1

1.2.3. Botulism
1.2.4. Black leg
1.2.5. Malignant edema (clostridial myonecrosis)
1.2.6. Infectious necrotic hepatitis (Black disease)
1.2.7. Bacillary haemoglobinuria
1.2.8. Enteric diseases associated with Clostridium perfringens
2. Some common chronic bacterial diseases of large animals
This portion gives attention to:-
2.1. Ulcerative lymphangitis
2.2. Tuberculosis
2.3.Paratuberculosis(John's disease)
2.4. Brucellosis
 3.Common disease of large animals caused by the member of in
the family of Pasteurellaceae
This portion gives attention to:-
3.1. Pasteurellosis
2
 Course cont’d---
 4. Some common bacterial diseases of large animals caused by
the member of in the family Enterobactericeae
This portion gives attention to:-
4.1. Colibacillosis
4.2. Salmonellosis
5.Diseases of large animals associated with order Actinomycetes
This portion gives attention to:-
5.1. Actinomycosis
5.2 Actinobacillosis
5.3. Dermatophilosis
6. Diseases of large animals associated with genus Mycoplasma
This portion gives attention to:-
6.1. Contagious Bovine Plueropneumonia (CBPP)
6.2. Contagious Caprine Plueropneumonia (CCPP)
 7. Other different bacterial diseases of large animals
This portion gives attention to:-
7.1. Infectious keratoconjuctivitis(group work) 3
 Course out line cont’d---
7.2. Listeriosis (group work)
7.3. Swine erysipelas( group work)
7.4. Leptospirosis(group work)
(group
work)
This portion gives attention to:-
1. Introduction to mastitis
2. Diagnostic methods of mastitis
3. Mastitis in cattle
4. Mastitis in domestic animals other than cattle
 5. Complication of mastitis
Part III. Some common viral diseases of large animals
1. Viral diseases of large animals characterized by nervous signs
This portion gives attention to:-
 1.1. Rabies
 2. Some common viral diseases of large animals characterized
by alimentary tract signs 4
 Course out line cont’d---
 This section gives attention to:-
2.1. Foot and Mouth Disease(FMD)
2.2. Rinder pest (Cattle plague)
2.3. Peste Des Petitis Ruminants (PPR)
2.4. Malignant Catarrhal Fever(MCF)
2.5. Bovine Virus Diarrhea(BVD)
3. Some common viral diseases of large animals characterized
by skin lesions
This section gives attention to:-
3.1. Lumpy Skin Disease(LSD)
3.2. Sheep pox and Goat pox
 3.3. Contagious Ecthyma( Contagious pustular dermatitis also
called orf)
3. 4. Papillomatosis(group work)
4. Some common vector born viral diseases of large animals
This portion gives attention to:-
4.1. Rift Valley Fever(RVF) 5
 Course cont’d---
4.2. Nairobi sheep disease
4.3. Blue tongue
 5. Viral disease of large animals characterized by respiratory
and nerve signs
This portion gives attention to:-
5.1. Maedi - visna
6. Viral diseases of swine characterized by the involvement of
the body as whole
This portion gives attention to:-
6.1. Classical swine fever(group work)
6.2. African swine fever (group work)

This portion gives attention to:-

4.1. Introduction to prions
4.2. Common diseases of large animals caused by prions
4.2.1. Bovine Spongiform Encephalopathy(BSE)
6
4.2.2. Scrapie
 Course cont’d---
Part V. Some common rickettsial diseases of large animals
This portion gives attention to:-
5.1. Heart water(Cowdriosis)
5.2. Anaplasmosis
Part VI. Common fungal diseases of large animals
This portion gives attention to:-
6.1. Dermatomycosis(Dermatophytosis also called Ring worm)
Part VII. Some common metabolic diseases of large animals
This portion gives attention to:-
7.1. Milk fever
7.2. Parturient paresis
7.3. Ketosis
7.4. Copper deficiency
7.5. Cobalt deficiency
7.6. Vitamin A deficiency
7.7. Hypomagnesaemia tetani
7.8. Selenium/vitamin E deficiency 7
I. Some common bacterial diseases of large animals
1. Anthrax
It has different names in different areas and named as Splenic
fever, wool sorter disease, Siberian ulcer, Charbon, Milzbrand.
Anthrax is one of the most dangerous infectious diseases of all
warm blood animals except birds.
It is soil born disease of haemothermic mammals including human
beings.
So that anthrax is zoonotic.
The disease is mostly characterized by its
 Acute form
 Septicemia
 Intoxication
 Formation of painful, edematous, subcutaneous swellings called
carbuncle that may appear about the throat, lower neck, floor of
the thorax and abdomen, prepuce, and mammary gland.
8
 Anthrax cont’d---
Etiology
Anthrax is caused by Bacillus anthracis.
Bacillus anthracis is gram positive aerobic spore forming rod
shaped bacteria.
The organism forms spores that help to persist in the environment
for decades.
B. anthracis forms spore in unfavorable environmental conditions
in the presence atmospheric oxygen in temperature of 15 – 42 0C.
That is why vegetative cells of B. anthracis in an unopened carcass
quickly die without sporulating.
Because of the rapid PH change following death and
decomposition of carcass.
The bacteria grows in common laboratory media like conventional
blood agar.
It forms capsule in infested organisms “ in vivo” and in artificial
media “ in vitro” that are rich in protein and it is the characteristics
of the virulent strains of B. anthracis. 9
 Anthrax cont’d---
Pathogenic strains have plasmid-encoded virulent factors that
include:-
 1. A poly D- glutamic( protein) capsule which aids in resistance to
phagocytosis and is encoded by virulence gen
 2. A tripartite toxin
A tripartite toxin comprised of:-
 Edema factor I
 Lethal factor II
Protective antigen factor III
So that protecting Antigen (PA) plus Edema Factor (EF) produce
edema toxin.
While protecting Antigen (PA) plus Lethal Factor (LF) produce
lethal toxin.
These toxins cause tissue swelling (edema) and death respectively.
NB. B. anthracis has been manufactured as a biologic warfare agent.
Spore form of B. anthracis is resistant to many disinfectants and to
desiccation. 10
 Anthrax cont’d---
That is why the spore can survive 60 – 100 years in the soil.
Epidemiology the disease
Anthrax has been reported from nearly every continent.
It is common in agricultural regions with neutral or alkaline soil.
But calcareous soils as acid soils reduce the survival of B.
anthracis.
In these regions, anthrax periodically emerges as epizootics among
susceptible domestic and wild animals.
These epizootics are usually associated with drought, flooding, or
soil disturbance, and many years may pass between outbreaks.
During interepidemic periods, sporadic cases may help maintain
soil contamination.

11
 Anthrax cont’d---
This area in tropics and sub tropics where there is a continuous
epidemics( epizootics) of anthrax called incubator area of B.
anthracis.
Although the area prevalence varies with the soil, the climate and
the efforts put into suppressing its occurrence.
It is often restricted to particular areas, the so-called “anthrax
belts” where it is enzootic.
The most susceptible animals are
Large and small ruminants
Equines and camel
Wild herbivores animals
Swines are less susceptible than the above species of animals.
 Dogs and cats are even less susceptible than swines and they
become sick when they are infested with large dose of spores.

12
 Anthrax cont’d---
Young animals are more susceptible than the older ones.
In epidemiology of anthrax we must understand
 1. Source of infection
 2. Mechanism of transmission
 3. Susceptible animals
 4. Risk factors of anthrax
1. Source of infection
These are sick animals that expel the organism together with faces
urine and together with different discharges.
The organism may also expel from sick animals together with
different exudates ( through necrotized carbuncle) and together
with black tarry blood through natural orifices at agony stage.
The organism can also expel from sick animal with
haematophages( as mechanical vectors).
Source of infection of anthrax can be contaminated soil with spore
of B. anthracis.
13
That is why anthrax is one of soil born diseases of animals.
 Anthrax cont’d---
2. Mechanism of transmission
The most dangerous factor for transmission anthrax is the carcass
of dead animals as it contains large amount of B. anthracis
organism.
B. anthracis can infect animals directly from
 The soil or from fodder grown on infected soil
 From contaminated bone meal or protein concentrates
 Infected excreta, blood, or other discharges from infected animals
Biting flies, mosquitoes, ticks, and other insects have often been
found to harbor anthrax organisms( mechanical vectors). Example
tabanid flies.
The most important disseminator of B. anthracis are scavengers
like vulture birds, street dogs and different wild carnivores.
3. Susceptible animals
Infection gains entrance to the body by
Ingestion
 Inhalation
 Through the skin( macro and micro wounds) 14
 Anthrax cont’d---
Most animals are infected by the ingestion of contaminated food or
water.
Injury to the mucous membrane of the digestive tract will facilitate
infection but there is little doubt that infection can take place
without such injury.
The increased incidence of the disease on sparse pasture is probably
due both to the ingestion of contaminated soil and to injury to the
oral mucosa facilitating invasion by the organism.
Inhalation infection is thought to be of minor importance in
animals, although the possibility of infection through contaminated
dust must always be considered “Wool sorter's disease” in humans
is due to the inhalation of anthrax spores by workers in the wool
and hair industries.
The organism may infect susceptible animals and human beings
through skin( micro and macro wounds) and cause cutaneous form
15
 Anthrax cont’d---
of anthrax with formation of carbuncle.
4. Risk factors of anthrax
1. Host risk factors
2. Environment risk factors
3. Pathogen risk factors
1. Host risk factors
The disease occurs in most domestic animals.
However, it is most common in cattle and sheep and less frequent in
goats and horses.
Humans occupy an intermediate position between these groups.
The relatively resistant animals are pigs, dogs, and cats.
2. Environment risk factors
Outbreaks originating from a soil-borne infection always occur
after a major climate change.
 For example heavy rain after a prolonged drought.
 Dry summer months after prolonged rain, and always in warm
weather. 16
Mode of transmission of B. anthracis

17
 Anthrax cont’d---
Other environmental risk factors include
Close grazing of tough
 Scratchy feed in dry times, which results in abrasions of the oral
mucosa
3. Pathogen risk factors
When material containing anthrax bacilli is exposed to the air,
spores are formed that protract the infectivity of the environment
for very long periods.
The spores are resistant to most external influences including the
salting of hides, normal environmental temperatures and standard
disinfectants.
Pathogenesis
After wound inoculation, ingestion, or inhalation, spores of the
bacteria is changed to vegetative cell.
The bacteria then multiply and enters to the lymphatic system and18
 Anthrax cont’d---
then to regional lymph nodes.
From lymph nodes the bacteria enter to blood system and captured
by phagocytic cells.
The organisms are resistant to phagocytosis, in part due to the
presence of the poly-o-glutamic acid capsule and distributed
through out the body of the organism especially in spleen.
There the bacteria are engulfed by macrophages and proliferate
inside them.
The bacteria then destruct the macrophages by lethal toxin and
enters to the blood system again.
In the blood circulation system, it reproduces and releases toxins
that helps to destruct tissues and cause intoxication.
If the infected organism is low resistant and the bacteria is high
virulent strain, septicemic form of anthrax may develop and death
occurs with in some hours after infection. 19
 Anthrax cont’d---
However if B. anthracis infest the animal through wounds of skin,
cutaneous form of anthrax( carbuncle) develops.
Carbuncle is edema( swelling) and is characterized by serous –
hemorrhagic inflammation of sub - cutaneous tissue due to toxin of the
bacteria found there.
Toxin may be absorbed from carbuncles and cause intoxication in
animals.
The bacteria may enter to regional lymph nodes again to cause
hemorrhagic lymphandinitis.
Finally from regional lymph nodes the bacteria may enter to blood and
to cause septicemic form of anthrax.
Clinical signs
The incubation period and the clinical course of the disease depends on
Resistance of the host
Route of infection
Dose and virulence of bacteria 20
 Anthrax cont’d---
Typically, the incubation period is 3 – 7 days (rarely it ranges 1−14
days).
The clinical course ranges from per acute to chronic.
There are two clinical forms of anthrax
1. Septicemic form
2. Carbuncle form
Depending on localization of pathological changes, anthrax has
Intestinal form
Cutaneous form
Pneumonic form
Angenose form(in pigs)
Such types of conditional classification helps us to understand the
clinical forms of the disease.
Septicemic form of anthrax can occur independently or as
21
secondary in carbuncle form of anthrax and vice versa.
 Anthrax cont’d---
Depending on the length of incubation period, anthrax usually has
the following forms
 1. Peracute
 2. Acute
 3. Subacute
 4. Chronic
1. Peracute form of anthrax
This form of anthrax is common in sheep and goat and rarely in
cattle and horses.
The peracute form anthrax is characterized by sudden onset of the
disease and a rapidly fatal course in sheep and goat.
Affected animals show staggering, dyspnea, trembling, collapse, a
few convulsive movements, and death may occur in sheep, or goats
with only a brief evidence of illness with out showing some
typical clinical sign of the disease.
However, at agony stage, bloody bubble is expelled through
mouth and nose.
Peracute form of anthrax in cattle and horses is characterized by
22
 Anthrax cont’d---
 Depression, dysponea and cyanosis.
Fever that can reach up to 41 – 42 0C.
 The animal may die within some minutes up to an hour at collapse.
2. Acute
This form of anthrax affects mostly cattle and horses.
It is characterized by
Fever
 Dysponea
 Shivering
 Inappetance
 Absence of rumination and pneumonia in cattle
 Colic in horses
 Rarely constipation or bloody diarrhea
 Haematuria 23
 Anthrax cont’d---
Finally the animal weakened, dysponea increased and cyanosis of
visible mucous membrane, rarely with petechial haemorrhages.
In this form of anthrax, swellings around pharynx, larynx of the
neck, sternum, lower abdomen, and external genitalia may occur.
Death usually occurs within 2 – 3 days of onset.
During period agony bloody bubbles of fluid is expelled through
mouth and nose.
3. Subacute
 Incubation period is 6 – 8 days
 Symptoms of the disease are similar to the acute form but they
develop slowly.
4. Chronic
 Incubation period is 2 – 3 months
 With clinical sign of progressive emaciation
 Chronic form anthrax can be suspected after slaughtering animals24
 Anthrax cont’d---
by observation of hemorrhagic infiltrate in intra madibular region
and by observation of infected intamadibular and
retropharyngeal lymph nodes during carcass inspection.
1. Carbuncle form anthrax: - It is characterized by formation
edematic swelling on different parts of the organism specially
around head, shoulder, sternum and abdomen.
Carbuncle is edematic swelling which is hard, hot and painful at
the beginning.
Then after some days it softened, cold and painless and at the
center it forms necrotic mass with holes through which the exudate
is expelled and contaminate the environment.
Carbuncle form of anthrax occurs on places where the bacteria
enter or as secondary in septicemic form of anthrax.
2. Intestinal form of anthrax: - This form of anthrax is common in
equines as secondary in septicemic form of anthrax and
25
 Anthrax cont’d---
characterized by colic, constipation followed by bloody diarrhoea.
3. Pneumonic form of anthrax: - It is characterized by progressive
hemorrhagic pneumonia and edematic lung.
4. Angenose form of anthrax: - This form of anthrax is common in
pigs and characterized by its chronic character, fever, angina and
pharyngitis.
The typical clinical signs that are seen in angenose form of anthrax
are
 Edema around neck
 Dysponea
 Difficulty in swallowing and coughing
 Sick pigs have abnormal chirp breathing sound
 Cyanosis of visible mucous membrane
 Edema of pharynx and larynx may lead the animal to die at hypoxia
26
Carbuncle form anthrax in horse and cow

27
Carbuncle form anthrax in cow

28
Cutaneous form of anthrax in human beings

29
Necrotized carbuncle in cutaneous form of anthrax in human
being

30
 Anthrax cont’d---
Necropsy findings
Do not open the carcass of animals if you suspect anthrax.
If the carcass is already opened by mistake and anthrax is
suspected on the way, please stop to open and think methods that
help you to control environmental contamination by Bacillus
anthracis spore.
The carcass of animals that are died at anthrax has the following
features
Rigor mortis is frequently absent or incomplete
All natural orifices usually exude dark, tarry blood that does
not clot.
 Carcass undergoes gaseous decomposition quickly and has marked
bloating assuming the characteristic “sawhorse” posture.
If a necropsy is carried out, there is
 Failure of the blood to clot and widespread ecchymosed,
bloodstained serous fluid in the body cavities,
 Severe enteritis and splenomegaly which are strong indications of
the presence of anthrax. 31
 Anthrax cont’d---
The enlarged spleen is soft, with a consistency likened to 'blackberry jam‘.
Subcutaneous swellings containing gelatinous material, and enlargement
of the local lymph nodes are features of the disease in horses and pigs.
Lesions are most frequently seen in the soft tissues of the neck and
pharynx in these species.
In pigs with chronic anthrax, the lesions usually are restricted to the
tonsils, cervical lymph nodes, and surrounding tissues.
The lymphatic tissues of the area are enlarged and are a mottled salmon to
brick-red color on cut surface.
Diphtheritic membranes or ulcers may be present over the surface of the
tonsils.
The area around involved lymphatic tissues generally is gelatinous and
edematous.
A chronic intestinal form involving the mesenteric lymph nodes is also
recognized.

32
Rapid bloated carcass of cow died at anthrax

33
Dark and tarry blood that oozes through natural orifices (nasal
cavity)during anthrax in cows

34
Dark and tarry blood that oozes through natural orifices (nasal
cavity)during anthrax in zebra

35
Splenomegaly during anthrax

36
Diagnosis
Diagnose of anthrax is done based on
1. The epidemiology of the disease
2. Clinical signs
3. Necropsy findings
The above diagnostic results give tentative diagnose of the disease.
4. Laboratory
Laboratory diagnose gives the confirmatory diagnose of the disease.
In epidemiological diagnose of the disease, consider
Species of animals that are affected in the area
 Sudden death of animals in pasture in endemic areas after
agricultural activities and flooding.
In clinical signs consider
Per acute and acute nature of the disease in most cases with
septicemia and/or carbuncle and angina on affected animals and
37
 Anthrax cont’d---
pigs respectively.
In necropsy findings, if there is a good reason to suspect the
existence of anthrax, the carcass should not be opened.
How ever, consider the presence of
 Incomplete rigor mortis
Oozing of dark, tarry blood through natural orifices that does
not clot.
 Rapid bloating of carcass
Tentative diagnosis of anthrax helps us to do necessary
prevention measures concerning human beings and the rest of
animals.
However the diagnosis must be confirmed by isolation and
identification the bacteria in laboratory.
In live animals, it is possible to collect aseptically
 Blood and fluid from peripheral blood vessels and local edematic
tissues respectively by needle puncture. 38
 Anthrax cont’d---
Prepare smears and stain with appropriate staining techniques( Gram
staining and/ or polychrome methylene blue).
In died animals to confirm the diagnosis on an unopened carcass
 Peripheral blood or local edema fluid should be collected by needle
puncture.
 Smears prepared from these fluids should be stained with polychrome
methylene blue and examined.
These fluid samples can also be used for bacteriological culture if smear
results are equivocal.
If decomposition of a carcass is advanced, collect
 A small quantity of blood or tissue from the fresh surface of an
amputated tail or ear in the direction where the animal lays.
A portion of spleen is the specimen of choice for bacteriological
culture if the carcass has been opened by mistake except from pigs.
 From dead pigs, edematic tissue, retropharyngeal and
interamandibullaris lymph nodes are the specimens of choice
39
 Anthrax cont’d---
NB. If anthrax is suspected, then shipping diagnostic samples via the
mail or courier systems is strongly discouraged.
Instead, samples should be appropriately packed, labeled and
transported directly by ice box with ice packs to the laboratory by
one of the staff members of the veterinary clinic.
Samples for confirmation of diagnosis
1. For bacteriology
1.1. From unopened carcass
 Blood or edema fluid in sealed, leak proof container
1.2. From opened carcass
Above samples plus spleen (local lymph nodes in horses, pigs) in
sealed, leak proof containers (direct smear and culturing)
2. For histology
 Formalin-fixed spleen or local lymph nodes if carcass has been
opened
NB. Note the zoonotic potential of this organism when collecting,
handling carcass and submitting specimens.
40
 Anthrax cont’d---
In laboratory the following activities are done
Staining by Gram and/ or
Polychrome methylene blue and capsule stainings( if the
specimens are fresh) and
Culturing on suitable media like blood agar
Serological tests like direct and indirect FAT
 The Ascoli test that can be used to demonstrate antigen in
severely decayed tissue.
PCR technique has been used in some referral laboratories
B. anthracis in polychrome methylene blue staining

41
 Anthrax cont’d---
NB. Protect your self from anthrax while you are taking specimens
by wearing protecting clothes , gloves, masks and by using
effective disinfectants.

NB. Since it is zoonotic disease all the procedures in the laboratory

must be done under biosafety cabinet using safety Bunsen
burner to control spattering of the pathogen while you flame the
inoculating loop.
42
 Anthrax cont’d---
Biosafety cabinet Safety Bunsen burner

43
B. anthracis in Gram and capsule stainings

44
 Anthrax cont’d---
Pure culture of B. anthracis on blood agar

45
Bacillus anthracis, medusa head colony morphology under
microscope

46
B. anthracis in spore staining

47
Ascoli ring precipitation test

48
Differential diagnosis
Anthrax must be differentiated from many animal diseases that
have similar clinical and path morphological manifestations like
Per acute black leg
Peracute blackleg
Malignant edema
Bacillary haemoglobinuria
Pasteurellosis
Enterotoxaemia in sheep
Piroplasmosis
Colic in equines
Tympany in ruminants
Hypomagnesemic tetany
Light and heat strike
49
Treatment
Treatment must be started with isolation of sick animals from the
herd.
The rest of animals should be moved to another pasture away from
where the bodies had lain and any possible soil contamination.
Administer for sick animals antibiotics and others like
 1. Penicillin ( 20 000 IU/kg BW twice daily I/M) for 3 – 5 days has
had considerable vogue, but streptomycin (8-10g/d) in two doses
intramuscularly for cattle) is much more effective.
 2. Oxytetracycline (5 mg/kg BW per day) parenterally(I/M) for 3 –
5 days has also proved superior to penicillin.
 3. Other antibacterial, including amoxicillin, chloramphenicol,
ciprofloxacin, , methicillin and netilmicin, doxycycline,
erythromycin, gentamycin, and sulfonamides and their combination
also can be used.
4. Specific hyper immune serum and Gamma globulin should also 50
 Anthrax cont’d---
administered for at least 5 days in doses of 100 - 250 mL S/C daily
if it is available.
Specific hyperimmune serum and gamma globulin give good
result if they are administered in combination with antibiotics.
NB. To use the specific serum for treatment of anthrax, it must be
warmed to 37 – 38 0C in water bath.
What makes difficult to use specific hyper immune sera for
treatment of different animal diseases in veterinary practice is in
that they are very expensive and cause anaphylaxis shock.
To prophylaxis anaphylaxis shock, administer 0.5 – 1 ml serum
S/C first and administer the rest of the dose after 15 – 30 minutes.
Control and prevention measures
Control measures
When an outbreak of anthrax occurs the following are part of the
animal disease control program that must be implemented as they
51
 Anthrax cont’d---
indirectly reduce human exposure.
These are
Placing of the farm in quarantine
 Destruction of discharges and cadavers
Vaccination of survivors
Prohibition of movement of milk and meat from the farm during
the quarantine period should prevent the entry of the infection into
the human food chain.
The methods that are used to control anthrax by destruction of
discharges and cadavers are
 1. Disposal of infected materials
 2. Disinfection of premises
1. Disposal of infected materials
It important and hygienic and it is one of the biggest single factor
in the prevention of spread of the disease.
Infected carcasses should not be opened but immediately burned
52
 Anthrax cont’d---
in situ or buried, together with bedding and soil contaminated by
discharges.
If this can not be done immediately, a liberal application of 5%
formaldehyde on the carcass and its immediate surroundings will
discourage scavengers.
Burning is the preferred method of disposal of the infected
materials
However, if it is impossible we can use burials.
Burials should be at least 2 m deep with an ample supply of
quicklime.
All suspected cases and in-contact animals must be segregated
until cases cease.
Thereafter the affected farm must be kept in quarantine to prevent
the movement of livestock.
The administration of hyperimmune serum to in-contact animals
may prevent further losses during the quarantine period,
However, prophylactic administration of a single dose of long 53
 Anthrax cont’d---
acting tetracycline or penicillin is a much commoner tactic.
2. Disinfection of premises
Disinfection can be carried out immediately before spore
formation can occur, ordinary disinfectants or heat (60°C (140 0F)
for a few minutes) are sufficient to kill vegetative forms.
However, when spore formation occurs (i.e. within a few hours of
exposure to the air), disinfection is almost impossible by ordinary
means.
So that strong disinfectants such as:-
5% Lysol require to be in contact with spores for at least 2 days.
 Strong solutions of 8 – 10 % of formalin or 5 – 10 % of sodium
hydroxide are probably most effective.
Additionally we can use the following different methods and
chemicals to sterilize materials.
Peracetic acid (3% solution) is an effective sporucidal and, if
applied to the soil in appropriate amounts (8 liter/m2).
Infected clothing should be sterilized by soaking in 10% 54
 Anthrax cont’d---
formaldehyde.
Shoes may present a difficulty and sterilization is most efficiently
achieved by placing them in a plastic bag and introducing
ethylene oxide for 18 hours.
 Hides, wool, and mohair are sterilized commercially by gamma-
irradiation
NB. Special care must be taken to avoid human contact with
infected material and, if such contact does occur, the contaminated
skin must be thoroughly disinfected.
Prevention measures
In livestock, anthrax can be prevented largely by
 Annual vaccination of all grazing animals in the endemic area and
 Implementation of control measures during epizootics
The non encapsulated strain of lyophilized attenuated live vaccine
is used almost universally for livestock immunization.
Vaccination should be done 2 – 4 weeks before the season when
outbreaks may be expected. 55
 Anthrax cont’d---
Dose of vaccine
 1 ml S/C for large animals
0.5 ml S/C for small animals
Immunity lasts for 1 year.
NB. Since anthrax vaccine is a live attenuated vaccine, antibiotics
should not be administered within 1 week of vaccination.
Other methods those are used to prevent anthrax are:-
 Livestock at risk should be immediately treated with a long-acting
antibiotic( prophylaxis dose) and/ or specific hyper immune serum
or Gamma globulin to stop all potential incubating infections.
 This must be followed by vaccination after 7 – 10 days of
antibiotic treatment.
 If any animals become sick after initial treatment and/or
vaccination should be retreated immediately and revaccinated a
month later.
 Simultaneous use of antibiotics and vaccine is inappropriate, as
available commercial vaccines for animals are live vaccines. 56
 Anthrax cont’d---
In addition to therapy and immunization, specific control
procedures are necessary to control and prevent its spread.
These include the followings : -
 Notification of the appropriate regulatory officials
 Rigid enforcement of quarantine (after vaccination, 2 weeks before
movement off the farm, 6 weeks if going to slaughter).
Prompt disposal of dead animals, manure, bedding, or other
contaminated material by cremation (preferable) or deep burial.
Isolation of sick animals and removal of well animals from the
contaminated areas
Cleaning and disinfection of stables, pens, milking barns, and
equipment used on livestock( intensive farms).
Use of insect repellents( intensive farms).
Control of scavengers that feed on animals dead from the disease.
Observation of general sanitary procedures by people who handle
diseased animals, both for their own safety and to prevent spread of
the disease. 57
 Anthrax cont’d---
 Contaminated soils are very difficult to completely decontaminate
but formaldehyde and other strong disinfectants will be successful
if the level is not excessive.
 The process generally requires removal of soil and burning or
burring of the contaminated soil together with dead animal carcass.
Incineration of the carcass of died animals at anthrax
Dig a pit about 2 feet deep and exceeding the length and breadth of
the carcass by about 1 foot on each side
Dig a trench 1 foot by 1 foot along the length of the center of the pit
extending beyond the ends of the pit by about 3 feet; this serves as
an air duct for the fire under the carcass .
Fill the trench and cover the bottom of the pit with straw and soak
them with an accelerant (kerosene or diesel fuel).
Cut heavy woods pallets to fit across the trench and within the
sides of the pit and place them on top of the straw.
Add other pieces of wood (or coal) until the pit is filled to the level
58
 Anthrax cont’d---
 Depending on the location of the carcasses and the environmental
rules in place and only in extenuating circumstances, tires have
been used in addition to or in place of wood (or coal).
Saturate all of this with accelerant.
 The carcass then can be lifted or drawn onto the pyre (combustible
heap).
 Pour further accelerant over the carcass and ignite the fire at either
end of the trench.
 Once the incineration is well under way (probably after the first
hour), cover the pyre with corrugated metal or other metal sheeting
to retain heat but not lose ventilation.
 If blood and body fluids have contaminated the ground and material
under the animal, they should be incinerated as well.
 Remove soil deep enough to collect any blood and body fluids that
have seeped into it.
 This could be up to 6 inches(15.3 cm). 59
 Anthrax cont’d---
Cross section of incinerator trench

60
Anthrax cont’d---

61
 Anthrax cont’d---
 This material can be placed on top of the carcass prior to igniting
the pyre.
If soil and other related materials cannot be incinerated, it can be
disinfected with 5% formaldehyde or NaOH solution by using 47
liters of solution per 0.85 square meter.
In Ethiopia it is difficult to practice incineration of the carcass as
above.
But it is possible to bury the carcass in 2 m depth pit under ground.
Selecting leveled area prepare the pit.
Put the carcass and all contaminates materials and soil cover with
Quick lime(Cao powder) and cover the soil.
The area must be free from grazing and agricultural activity and
well fenced.

62
2. Livestock diseases associated with clostridial infection
This portion gives attention to:-
 2.1.Introduction to the general characteristics of clostridial infection
2.2. Tetanus
2.3. Botulism
2.4. Black leg
2.5.Malignant edema(clostridial myonecrosis)
2.6. Infectious necrotic hepatitis (Black disease)
2.7. Bacillary haemoglobinuria
 2.8. Enteric diseases associated with Clostridium perfringens

63
 Clost. Cont’d---
2.1. Introduction to the general characteristics of
clostridial infection
The clostridia are of major importance in farm animals as primary
causes of disease.
They rarely act as secondary invaders except where gangrene is
already present.
Exotoxins are important in pathogenesis of most clostridia diseases.
But the potency of the exotoxins produced and the invasive ability
of the clostridia vary.
The toxins of the different organisms vary in their effects and in the
manner in which they gain entry to the circulation.
So the pathogenic species which produce potent exotoxin can be
divided in to four groups based on potency of toxin production and
invasive ability of the bacteria.
I. Neurotropic clostridia
This group of clostridia produces potent neurotoxin but they are
64
non invasive.
 Clost. Cont’d---
As the result the bacteria colonize the host to a very limited extent.
Example C. tetani and/or the toxin is ingested already preformed in
the feed example C. botulinum (type A - F)
Examples of animal diseases that are grouped under this group are
tetanus and botulism.
II. Histotoxic clostridia
They produce less potent exotoxin than the first group but are
invasive.
That means they elaborate in a more proper infection of the tissues.
Example are C. chauvoei, C. septicum, C. novyi, C. perfringens type
A and C
Animal diseases that are grouped under this group are:-
 Black leg
 Malignant edema(clostridial myonecrosis)
Infectious necrotic hepatitis (Black disease)
 Bacillary haemoglobinuria
65
 Clost. Cont’d---
III. Enterotoxaemic clostridia
Enterotoxins are formed in the intestines and absorbed from the gut
in to the blood stream and produce generalized toxemia.
Examples are C. perfringens type (A – E)
Animal diseases that are grouped under this group are enteric
diseases associated with Clostridium perfringens(A – E).
IV. Clostridia associated with antibiotic induced disease
They produce enteric disease that can be antibiotic induced.
Examples are C. difficile.
Animal diseases which is grouped under this group is enterocolitis
caused by Clostridium difficile.
2.2. Tetanus( Lock jaw)
It is an acute soil born and non-contagious wound infection of
animals characterized by increased reflexes in intensity.
So that the affected animals are easily excited into more violent by
sudden movement or noise that ends with tonic spasms upto
stiffness in all or in some groups of muscles caused by a specific66
 Tetanus cont’d---
neurotoxin produced .
Etiology
Tetanus in animals is caused by Gram positive bacterium which
produces terminal, spherical endospore that bulges the mother cell.
That is why the bacterium with its spore has a form of “Tennis
racket” or “Drum stick”.
The endospore of C. tetani is highly resistant.
However, autoclaving of the bacteria at 1210C for 15 minutes is
completely sporucidal.
Boiling kills the spore of most strains in 15 minutes.
The natural habitats of C. tetani are soil and faces.
The bacterium is motile by peritrichous flagella.
It is obligatory anaerobe and grow in Robertson’s cooked
medium(cooked meat medium) and on blood agar under
anaerobiosis.
Based on the flagella antigen there are 10 serological groups of C.
tetani . 67
 Tetanus cont’d---
But the neurotoxins produced by these serological groups is
uniform.
Epidemiology
Almost all mammals are susceptible to tetanus.
Dogs and cats are relatively more resistant than any other domestic
animals.
Birds are quite resistant.
Horses seem to be the most sensitive of all species, with the
possible exception of humans.
Young animals are more susceptible than old ones.
In young ruminants the case fatality rate is over 80%.
But the recovery rate is high in adult cattle.
How ever in horses it varies widely between areas.
In some areas almost all horses die acutely, in others the mortality
rate is consistently about 50%.
Source of infection
Source of infection are clinically healthy animals in which spore68 of
 Tetanus cont’d---
C. tetani is preset in their intestine and drops to the soil together
with their faces.
C. tetani organisms are commonly present in the feces of animals,
especially in horses, and in the soil is contaminated by these feces.
Spore of C. tetani can persist in soil for a long time.
That is why contaminated area always are the source tetanus
infection.
It is one of soil born disease.
Transmission
The portal of entry is usually through deep puncture wounds .
But the spores may lie dormant in the tissues for some time and
produce clinical illness only when tissue conditions favor their
proliferation.
Puncture wounds of the hooves are common sites of entry in horses.
Introduction to the genital tract at the time of parturition is the usual
portal of entry in cattle.
69
 Tetanus cont’d---
A high incidence of tetanus may occur in young pigs following
castration and in lambs following castration, shearing, docking,
vaccinations, or injections of pharmaceuticals preparates.
Neonatal tetanus occurs when there is infection in the umbilical
cord associated with insanitary conditions at parturition.
There is also what we call it 'idiopathic tetanus‘ occur
occasionally in young cattle without a wound being apparent.
It usually in association with the grazing of rough, fibrous feed, and
it is probable that toxin is produced in wounds in the GIT system.
Pathogenesis
The spores of C. tetani are unable to grow in normal tissue or even
in wounds if the tissue remains at the oxidation-reduction potential
of the circulating blood.
So that the presence of C. tetani in spore form in/ on different
tissues does not cause tetanus.
But instead the toxin that is produced cause the disease state.
70
 Tetanus cont’d---
Toxin production may occur immediately after introduction if and
only if : -
 The accompanying trauma has been sufficiently severe for
anaerobiosis creation or
 If foreign material has also been introduced to the wound and
creates anaerobiosis.
Other wise it may be delayed for several months until subsequent
trauma to the site causes tissue damage.
Suitable conditions for spore multiplication occur when: -
 There is tissues trauma due to different reasons and
 Small amount of soil or a foreign objects cause tissue necrosis and
create anaerobiosis.
Because they cause a lowering of the local tissue oxygen tension.
As the result the spores of C. tetani proliferate and produce battery
of toxins.
These are: -
71
 1. Protease,fibrinolysin and ribonuclease
 Tetanus cont’d---
 2.Tetanolysin
 3. Tetanospasmin (neurotoxin)
Hower in pathogenicity of C. tetani, tetanolysin and tetanospasmin
play an important role.
1.Tetanolysin
Tetanolysin is a cytolysin that increases the permeability of cellular
membranes through cell lysis.
It is heat and O2 liable and it can cause lysis RBC and tissues.
Tetanolysin promotes local tissue necrosis.
2. Tetanospasmin (neurotoxin)
It is plasmid - coded and responsible for the clinical signs of
tetanus.
Tetanospasmin is heat and oxygen stable.
Tetanospasmin is the cause of tetanus and is sometimes referred to
as tetanus neurotoxin (TeNT), as it acts on the central nervous
system.
It is highly lethal toxin ( 1×10 -7mg dose kills the mouse within 172–
 Tetanus cont’d---
– 2 days and easily neutralized by antitoxin).
The endospores of C. tetani enter to susceptible animal through
traumatized tissue created by different reasons like
 1.Surgical wounds( after castration or docking), shearing,
vaccinations, or injections of pharmaceuticals preparates if they are
not performed aseptically and antiseptically.
2. Via umbilicus or into the uterus following dystocia.
3. Through wounds of mechanical trauma and biting etc
The presence of facilitative anaerobiosis by different factors like:-
1. Deep wounds
2. The presence of necrotic tissue and hematomas
3. Mixed infection of wound with aerobic bacteria
These conditions create anaerobic condition and C. tetani spores
start to germinate.
Due to the multiplication of vegetative cells at the entry site, they
produce the potent exotoxin called tetanospasmin.
73
 Tetanus cont’d---
The toxin will be absorbed and travels to CNS( brain and spinal
cord) by peripheral nerves or blood stream and adsorb on motor
neurons.
The toxin acts in synapse mainly on post synaptic membrane.
 You know neurotransmission is controlled by the balance between:
 Excitatory( production of mediator called acetylcholine) and
 Inhibitory neurotransmitters( acetylcholine esterase).
Tetanospasmin blocks the inhibitory of neurotransmitters that
helps in depolarization of the postsynaptic membrane in conduction
of the electrical signal.
In the absence of inhibitory neurotransmitters, excitation of the
neuroaxon is unrestrained.
So that there is a continuous polarization of post synaptic
membrane.
The toxin binds to gangliosides in nerve tissue and once bound can
not be neutralized by antitoxin.
74
Mechanism action of tetanospasmin in synapses of motor
neurons

75
 Tetanus cont’d---
Tetanospasmin affect the pharynx, larynx, diaphragm and
intercostal muscles.
It also paralyzes the heart and breathing centers.
All the above conditions cause the death of affected animals at
asphyxiation and failure in blood circulation.
Clinical signs
The incubation period varies between 3 days and 4 weeks but
usually averages 10 – 14 days.
The rectal temperature and pulse rate are within the normal range
in the early stages.
But may rise later when muscular tone and activity are further
increased.
Tetanus usually has acute form.
Generally, there are two types of clinical manifestation of tetanus.
These are: -
 1. Ascending tetanus
2. Descending tetanus 76
 Tetanus cont’d---
1.Ascending tetanus
The toxin affects the muscle that are found around the wound
infection travels up through nerve fiber and tetanspasm develops
in the muscle of affected limb.
Then it spreads to the opposite limb and moves upwards.
This type of tetanus is called ascending tetanus.
Ascending tetanus is common in tetanus less susceptible animals
like dogs and cats.
2. Descending tetanus
If the tetanospasmin toxin circulating through blood stream, it will
affect the motor neuron of CNS.
As the result muscle spasm will start at head, maxillary, madibular
region then distribute to the neck and spinal cord lastly to limbs.
This type of tetanus is called descending tetanus and common in
high tetanus susceptible animals like human being and horses.
Clinical findings are similar in all animal species initially, there is
an increase in muscle stiffness, accompanied by muscle tremor. 77
 Tetanus cont’d---
The main clinical signs are:-
 Trismus with restriction of jaw movements because of spasm of
head muscles.
 Spasms of head muscles cause difficulty in prehension and
mastication of food, hence the common name of tetanus is lockjaw.
 In horses, the ears are erect, the tail stiff and extended, the anterior
nares dilated, and the third eyelid prolapsed
 Walking, turning, and backing are difficult
 Spasms of the neck and back muscles cause extension of the head
and neck.
 Stiffness of the leg muscles causes the animal to assume a
“sawhorse” posture.
 Uneven muscular contractions may cause the development of a
curve in the spine and deviation of the tail to one side.
 The reflexes increase in intensity and the animal is easily excited
into more violent by sudden movement or noise.
78
 Tetanus cont’d---
 An anxious and alert expression contributed to by an erect carriage
of the ears
 Retraction of the eyelids and dilation of the nostrils, and
and hyperesthesia with exaggerated responses to normal stimuli.
 Sweating is common
 General spasms disturb circulation and respiration which results in
increased heart rate, rapid breathing and cyanosis of visible
mucous membrane.
 Sheep, goats and pigs often fall to the ground and exhibit
opisthotonos.
 The animal may continue to eat and drink in the early stages but
mastication is soon prevented by tetany of the muscles, and saliva
may drool from the mouth.
 If food or water are taken, attempts at swallowing are followed by
regurgitation from the nose and may cause aspiration pneumonia.
 Constipation is usual and the urine is retained, partly as a result of
79
inability to assume the normal position for urination.
 Tetanus cont’d---
Tetanus in horses and ruminants with different clinical
manifestations

80
Tetanus in horses with stiffness and extended head and neck

81
Tetanus cont’d---

82
Tetanus in cattle
Stiffness of muscle with curved spine

83
Tetanus in dogs

84
Tetanus in dogs

85
Necropsy findings
There is no typical necropsy findings for tetanus, however, you
may observe
Complete rigor mortis
Black coloured un clotted blood
 Ecchymotic haemorrhage in different muscles
Haemorrhage on epicard, myocardium and pleura
Dilation of heart and edema of lung
Diagnosis
Diagnose is done based on:-
The clinical signs and history of recent trauma
Laboratory to confirm the diagnosis
The specimens of choice to confirm tetanus are
Exudates from deep wounds
Pieces of tissues in affected deep wounds
Pieces of liver and spleen
Additionally you can prepare smear from the above specimens and
86
Toxin identification
Control mouse Protected mouse

87
 Tetanus cont’d---
send to laboratory
 Blood ( 10 ml) in vacuutainer tube with out anticoagulant to extract
serum
Confirmatory diagnosis to tetanus is by two directions
 By demonstrating the presence of tetanus toxin in serum by
injecting to laboratory animals ( mice or guinea pigs)
 By isolation and identification of the bacteria and collection of the
toxin from isolated bacteria followed by toxin demonstration in
laboratory animals( mice and guinea pigs).
Differential diagnosis
Tetanus must be differentiated from:-
1. In all species of domestic animals
 Strychnine poisoning
 Meningitis
2. In horses
 Hypocalcemic tetany (eclampsia)
 Acute laminitis 88
 Tetanus cont’d---
 Hyperkalemic periodic paralysis
 Myositis, particularly after injection in the cervical region
3. In ruminants
 Hypomagnesaemia
 White muscle disease
 Polioencephalomalacia
 Enterotoxemia
Treatment
The main principles in the treatment of tetanus are to:-
Neutralize residual toxin
Eliminate the causative bacteria
Providing symptomatically treatment(control muscle spasms until
the toxin is eliminated or destroyed)
Provide supportive treatment
Tetanus antitoxin is administered but is of little value once signs
have appeared.
89
 Tetanus cont’d---
In the early stage of tetanus residual toxin is neutralized by IV
administration of antitoxin.
The recommended dose is controversial but for optimal results
horses should receive 300 000 IU IV every 12-hourly in three
injections.
Small and young animals 120000IU IV every 12 hours in 3
injections
Local injection of some of the antitoxin around the wound is
advised.
Good results have been obtained in horses by injecting 50,000 IU of
tetanus antitoxin directly into the subarachnoid space through the
cisterna magna.
NB. Tetanus antitoxin neutralize only the non – adsorbed tetanus
toxin that are found freely in the blood. So that early treatment of
tetanus with antitoxin is effective.
Such therapy should be supported by eliminating the causative
agent, symptomatically therapy(controlling muscle spasms) and90by
 Tetanus cont’d---
providing supportive treatment.
The causative agent is eliminated by:-
 Surgically opening of the abscess and removing of the pus and
necrotic tissues
 Washing of the infected wound with oxidizing antiseptics like 5%
iodine texture, 3% phenol(carbonic acid), 1:1000 Kmno4 and 3 –
5% H2o2 .
 Parenteral administration of penicillin and broad spectrum
antimicrobial agents( streptomycin, tetracycline, chloramphenicol
etc with their appropriate doses to control vegetative form of C.
tetani and other secondary infections.
The above treatment must be supported by providing
symptomatically treatment like:-
 1. Administration of curariform agents, tranquilizers, or barbiturate
sedatives for relaxation of the muscle tetany like
 Chlorpromazine (0.4-0.8 mg/kg body weight (BW) intravenously or
1.0 mg/kg BW intramuscularly, three or four times daily). 91
 Tetanus cont’d---
 Acetyl promazine (0.05 mg/kg BW twice daily) administered until
severe signs subside, are widely used or
 A combination of diazepam (0.01-0.4 mg/kg) and xylazine (0.5-1.0
mg/kg, intravenously or intramuscularly) for 3 or 4 times daily
until sever sign subside.
Beside the above methods, supportive treatment must be be given
like
 The horse should be placed in a quiet, darkened box stall with
feeding and watering devices high enough to allow use without
lowering the head.
Slings may be useful for horses having difficulty standing or rising.
 Administration of enemas with glucose that have 400 gm/ liter of
water.
 Periodically removing of faces from the rectum and massage or
catheterization of bladder may relieve the animals’ discomfort.

92
 Tetanus cont’d---
Control and prevention
Control and prevention of tetanus comprises using one or in
combination of the following methods:-
 1. Prevention of trauma, in time cleaning and disinfection of
wounds with appropriate antiseptics
2. Passive immuniazation
3. Active immunization
1. Prevention of trauma, in time cleaning and disinfection of
wounds with appropriate antiseptics
Proper skin and instrument disinfection is necessary during:-
Surgical operations
 Your help in dystocia cases
 Cleaning of the umbilicus of a new born animals
Castration, docking and shearing time
So that surgical procedures should be conducted with the best
possible aseptic techniques.
93
 Tetanus cont’d---
After surgery, animals should be turned out on clean ground,
preferably grass pastures.
Use only the oxidizing disinfectants that have iodine,
permanganates, and chlorine or peroxides as they dependably kill
the spores.
2. Passive immuniazation
Short-term prophylaxis can be achieved by the injection of tetanus
antitoxin.
The immunity is transient and persisting for only 10 - 14 days.
In high-risk areas it is important to vaccinate the animals with
tetanus toxoid; 1 month before mass castration of animals.
Tetanus antitoxin( 3000 – 5000 IU IV) together with broad
spectrum antibiotics should be given to any animal with a
penetrating wound, deep laceration or burning and/ or during
dystocia and the wound should also be cleaned aggressively with
appropriate antiseptics.
1. Tetanus antitoxin is often routinely given to mares following 94
 Tetanus cont’d---
foaling in dose of1500IU IV and to newborn foals in dose of 200IU
IV.
2. On farms where the incidence of tetanus in lambs is high,
antitoxin is usually given at the time of docking or castration; 200
IU IV has been shown to be effective.
3. The risk for tetanus in calves is lower than in lambs and tetanus
antitoxin is not commonly given at the time of castration.
3. Active immunization
Active immunization can be accomplished with tetanus toxoid.
Tetanus toxoid is formalin-inactivated adjuvanated toxoid which
induces long lasting immunity.
Primary vaccination requires two doses with 3 - 6 weeks apart.
Protective titer is obtained within 14 days of the second injection
and last for at least a year and more.
In high-risk areas, foals must be vaccinated with tetanus toxoid at
the age of 6 months.
Although immunity lasts longer than 1 year, it is common to 95
 Tetanus cont’d---
revaccinate horses yearly with a single booster injection.
Pregnant mares should receive a booster injection 4 - 6 weeks
before foaling to provide adequate colostral immunity to the foals.
Ewes are immunized with a similar schedule.
A prelambing booster vaccination is given yearly.
Commonly, commercial vaccines for sheep also contain antigens for
other clostridial diseases like black disease, enterotoxaemia and
bradsot for which sheep are at high risk.
Vaccination of cattle is usually not considered unless an outbreak of
the disease has occurred in the immediate past and further cases
may be anticipated.
2.3. Botulism
Botulism is a rapidly fatal motor paralysis caused by ingestion of
the neurotoxin produced by Clostridium botulinum types A - G.
Etiology
The causative organism Clostridium botulinum, rod shaped , Gram
positive spore-forming anaerobe, produces neurotoxins during 96
 Botulism cont’d---
vegetative growth.
C.botulinum forms sub-terminal spore in unfavorable environmental
conditions.
Spores can survive in the environment for over 30 years.
The spore-forming anaerobic organism proliferates in decomposing
animal tissue and sometimes in plant material.
Under favorable conditions of warmth and moisture the spores
germinate and vegetative cells multiply rapidly elaborating a stable
and highly lethal toxin called botulin.
Botulin is high potent biological toxin, it is more toxic than
strychnine.
C. botulinum can be cultured in laboratory on conventional
medium like blood agar and in anaerobic media like cooked meat
broth with thioglycollate(Robertson cooked meat medium) under
anaerobiosis.
Seven antigenically distinct toxin types (A - G), some with
subtypes, have been identified. 97
 Botulism cont’d---
These are neurotoxin A, B, C (C1 and C2 ), D, E, F and G .
Types A, B, E and rarely type F are most important in people.
Type C1 is important in most animal species, notably wild ducks,
pheasants, chickens, cattle, and horses; and D in cattle.
Types C and D are more common in warm climates like Africa.
The organism is present in the alimentary tract of animals that have
recently ingested contaminated material and may be introduced into
new areas in this way, or by birds and blowflies.

98
 Botulism cont’d---
The seven antigenically distinct toxin of Clostridium botulinum toxin
,susceptible hosts and their sources

Types Toxin(S) Most susceptible Source of toxin

produced animals C. botulinum
A A Human and chicken Vegetables, fruits, meat and
fish
B B Human rarely cattle, Meat and meat products (
horses and chicken often from pigs)
Cα C1(C2) Water fowl Invertebrate carcass and
rotting vegetation
Cβ C2(C1) Cattle , horses and Carcass , baled silage, chicken
dogs(human) manure as feed supplement
and spoiled feeds
D Mostly D Cattle, sheep ( Eating of contaminated bones
horses, humans) and carcasses of small
mammals due to calcium and
phosphorus deficiency. 99
 Botulism cont’d---
The seven antigenically distinct toxin of Clostridium botulinum toxin
,susceptible hosts and their sources
Types Toxin(S) Most susceptible Source of toxin
produced animals C.
botulinum

E E Humans Fish, fish products

and other foods
F F Humans Meat and fish
C. argentinense G Humans Soil

100
Epidemiology
Botulism has no geographical limitations in its distribution.
It affects most domestic animals, birds and human beings.
The most resistant animals to botulism are dogs, cats and pigs.
Source of infection
Most incidents of botulism are associated with the ingestion of
preformed toxin (forage botulism).
1. Toxin in feeds may result from the primary growth of C.
botulinum in the feed.
Examples are:-
Silage and hay may spoil to a stage and suitable for the growth of C.
botulinum.
Proliferation of the organism in decaying vegetable material. or
 2. From contamination of the feed with toxin containing carrion
forage called carrion-associated botulism( botulism with dead
carcass thrown to pasture)
 Carrion-associated botulism( botulism with dead carcass thrown to
pasture) can be:- 101
 Botulism cont’d---
 Either direct carrion ingestion that can be occurred due to calcium
and/ or phosphorus deficiency
From the contamination of the feed with toxin containing carrion.
3. Poultry manure and ensiled poultry litter have caused outbreaks of
botulism when used as cattle feed, as has poultry litter used for
bedding cattle.
It is probable that the source of toxin in poultry litter is from poultry
carcasses.
Pathogenesis
Botulism is potentially fatal intoxication associated with ingestion
of preformed toxin in foodstuffs.
Spores are present on food when gathered and processed.
If reliable temperature and pressure are not achieved air will be
evacuated but spores will remain.
This creates anaerobic and other favorable conditions that favor
spore germination and vegetative growth.
102
 Botulism cont’d---
As the result a potent toxin called botulin is released.
The neurotoxin( botulin) is absorbed from the intestinal tract and is
transported via the blood stream to peripheral nerve cells where it
binds to susceptible cells.
The heavy chain of the toxin is responsible for binding to the
receptors and translocation into the cell.
While the light chain of the toxin for resultant blockade of the
release of acetylcholine at the neuromuscular junction called
synapse.
Flaccid paralysis develops and the animal dies at respiratory
paralysis.
There are less common methods of acquisition of toxin botulin.
These are
1.Wound botulism
2. Intraintestinal toxicoinfection
1. Wound botulism
In wound botulism the spores are introduced into wounds where 103
 Botulism cont’d---
they germinate.
It is recorded in horses following castration, with omhalophlebitis,
umbilical hernias, with an infected wound and in association with
an injection abscess.
Due to the presence of necrotic tissues and hematomas creates
anaerobiosis and the spore germinates and release exotoxin at
localized site and spreads through out the body.
2. Intraintestinal toxicoinfection
In intraintestinal botulism the spores are introduced into intestine
where they germinate.
Due to the presence of necrotic tissues and hematomas in
intestinal lumen creates anaerobiosis and the spore germinates and
release exotoxin at localized site and spreads through out the body.
The disease is prevalent in horses specially in young horses.
The disease in foals also called the “shaker foal syndrome”
It is a disease of young foals up to 8 months of age with the highest
prevalence in foals 3 - 8 weeks of age. 104
 Botulism cont’d---
Clinical signs
The clinical signs of botulism are caused by:-
Flaccid muscle paralysis and progressive motor paralysis
 Disturbed vision, difficulty in chewing and swallowing
Generalized progressive weakness
 Death is usually due to respiratory or cardiac paralysis.
Incubation period of botulism is 18 hours upto 16 days.
It has peracute, acute, sub acute and chronic forms.
The length of incubation period and its forms depends on:-
The dose of botulin toxin
Species of animals and
At natural resistance of individual animals
The incubation period being shorter as the amount of toxin available
is increased.
Depending on the length of incubation period, botulism has the
following forms
1. Peracute 105
 Botulism cont’d---
2. Acute
3. Subacute
4. Chronic
1.Peracute form
In this form of botulism, animals die without prior signs of illness,
although a few fail to take water or food for a day before hand.
The disease is not accompanied by fever and the characteristic
clinical pictures is progressive symmetric muscular paralysis
affecting particularly the limb muscles and the muscles of the jaw,
tongue, and throat.
2. Acute form
In this form of botulism the animals show signs of
 Restlessness and incoordination
 Stumbling, knuckling and ataxia are followed by inability to rise or
to lift the head
Mydriasis and ptosis occur early in the clinical course
 Skin sensation is retained 106
 Botulism cont’d---
Shallow and fast breathing
Cyanosis of visible mucous membrane
Paralyze of the lower jaw and pharynx
In some cases the tongue becomes paralyzed and hangs
from the mouth
 The animal is unable to chew or swallow and it drools saliva
Atony and constipation
Difficulty in urination
 Affected animals lie in sternal recumbency with the head on the
ground or turned into the flank, like the posture of a cow with
parturient paresis but the cows do not respond to calcium therapy.
Fatality rate may reach 90 – 95%
3. Subacute form
The animal can stand and lie by it self
However quickly weaken while it moves
It likes to lie more time on the ground
107
 Botulism cont’d---
Mastication and swallowing is disturbed
Disturbance of swallowing results in aspiration of feed that can be
complicated by aspiration pneumonia or gangrene of lung.
Tones of heart are weak
4. Chronic form
Most of the physiological parameters are at the norm.
However, the affected animal spends most of it time by laying on
the ground.
And there is a progressive emaciation
Affected animals recover better than the other forms.
Necropsy findings
No characteristic gross and histological lesions develop, and
pathologic changes may be ascribed to the general paralytic action
of toxin.
But some pathological changes may help us to suspect botulism.
These are
The presence of unusual feedstuffs( bone, hair and pieces of 108
 Botulism cont’d---
of stones) in the forestomachs or stomach of animals
Pulmonary edema and pneumonia or gangrene of lung
 Congestion and excessive pericardial fluid which contains free-
floating strands of fibrin.
Hemorrhagic inflammation of the mucous membrane of intestine
 The presence of solid faces which is covered by mucous in the
rectum
Diagnosis
Diagnose of botulism is made based on
Epidemiology of the disease
Clinical signs
Necropsy findings
Laboratory
The first 3 methods give tentative diagnosis of the disease.
While laboratory diagnose is a confirmatory diagnose
Laboratory confirmations is attempted by:
109
 Botulism cont’d---
Detection of preformed toxin in serum, intestinal tract contents, or
feed
 Demonstration of spores of C. botulinum in the feed or
gastrointestinal contents
 Detection of antibody in recovering or clinically normal at-risk
animals.
Detection of preformed toxin in serum, intestinal tract contents, or
feed is done by toxin neutralization test on mice or guinea pigs( first
with polyvalent antitoxic serum then with monovalent antitoxic
serum).
Demonstration of spores of C. botulinum in the feed or
gastrointestinal contents can be done by inoculating the macerated
and heated( 65 – 80 0C) specimens on/ in suitable medium under
anaerobiosis.
The detection of antibody in chronically affected animals and at-risk
herd mates by an ELISA test has been used to support
110
 Botulism cont’d---
diagnosis in outbreaks of specially type C and type D botulism.
Differential diagnosis
Botulism must be differentiated from
Peracute form of anthrax
Rabies
Pseudo rabies( Aujeszky's disease)
Nervous form of Listeriosis
Marek’s disease
Poisoning of animals by poison plants and organophosphate/
/carbamate poisons
Parturient paralysis
Infectious equine encephalomyelitis
Treatment
Treatment of botulism is successful only in sub acute and chronic
form of the disease.
It can be treated using
111
 Botulism cont’d---
 1. Specific or polyvalent antiserum which are available in some
countries
They are effective if they are administered early in the course at a
dose of 30 000 IU for a foal and 70 000 IU for adult horses IV
A single dose is sufficient but it is expensive.
2. Supportive treatment like
Washing of stomach by 2 – 3 % NaHCo3
 Administration of warm enemas
 Administration of osmotic purgatives like 2% MgSo4 or Na2So4 or
laxatives
 Muzzling may be required to prevent aspiration pneumonia and
frequent turning to prevent muscle necrosis and decubital ulcers.
 Bladder catheterization may be required in horses that do not
urinate
 It is possible to administer broad spectrum antimicrobial drugs to
treat secondary complications such as during aspiration pneumonia.
112
 Botulism cont’d---
Prevention and control measures
Avoid contamination of animal feeds with soil, carcass of animals
and poultry manure and litter while you are preparing or storing
them.
Decaying grass or spoiled silage should be removed from the diet
Any dietary deficiencies in range animals should be corrected
specially calcium, phosphorus and/ or protein deficiency.
Hygienic disposal of carcasses is advisable to prevent further
pasture contamination
In area where botulism is endemic in poultry farms, stop to use
poultry manure and litter as feed of animals and as fertilizer in
pasture.
 Immunization of animals with toxoid that are prepared from toxins
of circulating types has practical importance for example
Vaccination with type-specific or combined (bivalent C and D)
toxoid is practiced in enzootic areas in Australia and southern Africa
113
 Botulism cont’d---
and give good result.
 Type B and C toxoid vaccine would be more appropriate for
prevention of disease in North America and Europe.
2.4. Black leg (Black quarter)
It is non - contagious, acute and febrile disease of cattle and rarely
sheep and goats caused by Clostridium chauvoei characterized by
emphysematous swelling with crepitating sound, usually in
heavy muscles that is why in some books they call it clostridial
myositis.
Etiology
Black leg is caused by Clostridium chauvoei.
Clostridium chauvoei is a gram positive, spore- forming, rod-
shaped and strictly anaerobic bacterium.
Spores are formed in aerobic conditions and are located central or
subterminal.
The size of spores bulges the mother cell.
The spores are highly resistant to environmental changes and 114
 Black leg cont’d---
disinfectants and persist in soil for many years.
Additionally the spores can be found faces of animals, dirty water
sources and in the intestine of animals.
The bacteria can be cultured on blood agar and anaerobic media
like cooked meat broth with thioglycollate(Robertson cooked meat
medium) under anaerobiosis.
Epidemiology
The disease can be endemic in particular areas, especially when
they are subject to periodical flooding.
Outbreaks of blackleg have occurred in cattle onfarms in which
recent excavations have occurred or after flooding.
Black leg is one of soil born infection.
Cattle, buffalo and rarely sheep and goats are susceptible to black
leg.
Cattle of all age groups are susceptible.
However, black leg very often affects cattle at the age of 6
months up to 4 years and animals with good body condition. 115
 Black leg cont’d---
In black leg enzootic areas, the disease never affects calves up to
3months of age and most cattle greater than 5 years old because of
colostral immunity and immunization subinfection respectively.
Occurrence of black leg is more at the end of winter season up to
the beginning of summer in our country this is due to
 Agricultural activities and flooding
 Increasement of the mechanical vector insects
 The season is favorable for animals to have good body conditions
 Mechanical damages of the mucousa of GIT by strong feeds
 The presence of high incidence of muscle trauma of animals
In enzootic area of blackleg, the case fatality rate approaches 100%.
Source of infection
Sick animals are source of infection.
 Blackleg infection can be endogenous as the organism may deposit
in muscle and other tissues (spleen, liver, and alimentary tract)
116
Means of transmission
Black leg is transmitted by soil, feed, pasture and water that are
contaminated by spores of Clostridium chauvoei.
Carcasses of animals that are dead because of black leg are also
very important means of transmission.
Insects can also transmit black leg.
Pathogenesis
In natural condition the organism can infect through two routes
 By ingestion and the microbes pass through the wall of the GI tract
 Through damaged skin
Then after the microbes gain an access to the bloodstream, are
deposited in muscle and other tissues (spleen, liver, and alimentary
tract) and may remain dormant. Or
It may sediment on muscle tissues including cardiac muscle and
spores start to multiply there as the muscle is rich in glycogen.
Macro and microtrauma on muscle facilitates the multiplication of
bacteria.
117
 Black leg cont’d---
As the result the bacteria produce exotoxins and aggressins.
The exotoxin and aggressins destruct the blood vessels and tissues
and develops inflammation.
Inflammatory exudate around the affected tissue contains bubble of
gas called H2S.
That is why carbuncle which is formed on affected muscles always
crepitate during palpation.
The exotoxins and other different toxins(that are produced from
damaged tissues) can be absorbed into blood stream and cause
intoxication of animals.
Intoxication of animals results in
Fever
Cardiovascular disturbance
Disturbance of liver functions
All these conditions lead the animal to die.
118
 Black leg cont’d---
Clinical signs
Incubation period of black leg is 1 – 2 days and rarely it can reach 5
days.
Very often black leg occurs in acute form with clinical signs like
Fever that may reach 41 – 42 0C.
 However, by the time clinical signs are obvious, body temperature
may be normal or subnormal.
Acute, severe lameness
 Characteristic edematous and crepitant swellings called
emphysematous carbuncle develop in the hip, shoulder, chest, back,
neck, or elsewhere.
At first, the swelling is small, hot, and painful.
 However, as the disease rapidly progresses, the swelling enlarges
and crepitate on palpation.
 The skin becomes cold and insensitive with decreased blood
supply to affected areas and it may have red - black colour.
 Swelling of regional lymph nodes. 119
 Black leg cont’d---
As infection progresses, the affected animals
 Depressed and anorexic with absence rumination,
 Fast breathes and tachycardic with weak pulse(100 – 120/ minute)
Prostrate and tremor and death occurs within 12 – 48 hrs.
In some cattle, the lesions may restrict to deep muscles, to the
myocardium and diaphragm and becomes difficult to observe
crepitation sound clinically.
Black leg can also affect tongues of animals.
Rarely black leg may occur peracute( septicemic) form in young
animals(3 – 5 months age) and in few cattle and usually: -
 The onset is sudden, and a few animals may be found dead
without premonitory signs.
In small ruminants black leg occurs similar to cattle but
emphysematous carbuncle not always appear and have the
following clinical signs
 Depression
120
Heifer dead because of black leg

121
 Black leg cont’d---
 Anorexia
 Lameness when the muscles of legs are affected
NB. Black leg in old cattle may appear atypically with clinical
signs like inappetance, depression and pain around affected
muscles with out the formation of emphysematous carbuncle.
Necropsy findings
If black leg is suspected by using epidemiological and clinical
diagnose, it is not advisable to open the dead bodies.
Cattle found dead of blackleg are often in a characteristic position
and it is easy to distinguish because
 The dead cattle is lying on the side with the affected hind limb
stuck out stiffly.
The necropsy findings are
 Bloating and putrefaction occur quickly and bloodstained froth
exudes from the nostrils and anus which clots rapidly.
 Hypoderm skin that is found on the of the affected muscles
hemorrhagic and have bubbles of gas. 122
 Black leg cont’d---
 Incision of the affected muscle mass reveals dark red to black,
swollen tissue with a rancid odor and thin, sanguineous fluid
containing bubbles of gas.
 Regional lymph nodes enlarge and have dark red colour in incision.
The thoracic cavity and the pericardial sac may contain excess
bloodstained fluid with variable amounts of fibrin.
Liver enlarges with foci of necrosis
Spleen enlarges and it is soft
Myocardium is found in parenchymatous degeneration.
The lungs are usually congested and may be atelectatic as a result of
abdominal tympany.
In sheep the muscle lesions are more localized and deeper and the
subcutaneous edema is not so marked, except around the head.
Gas is present in the affected muscles but not in such large amounts
as in cattle.
However, when the disease has resulted from infection of skin
123
wounds, the lesions are more obvious superficially, with
 Black leg cont’d---
Affected muscle and heart

124
 Black leg cont’d---
subcutaneous edema and swelling and involvement of the underlying
musculature.
NB. The heart and all skeletal muscles, including those of the
tongue, diaphragm, and lumbar region, must be checked, as the
lesion may be small and escape cursory examination.
Diagnosis
Diagnose of black leg can be done based on epidemiological,
clinical, necropsy findings and laboratory findings.
Specimens of choice to isolate C. chauvoei are
 Pieces of affected muscle tissues
 Liver
 Spleen
It is also possible to prepare air dried impression smears from
freshly cut lesions.
Collect samples from suspected muscle lesion for histopathology
The specimens are used for culturing and for FAT (direct and
indirect). 125
 Black leg cont’d---
NB. All specimens must be collected as soon as possible but not
after 2 – 3 hours of death and submitted to laboratory in air tight
container.
Differential diagnosis
Black leg must be differentiated from animal diseases like
Edematic carbuncle form of anthrax
 Malignant edema( false black leg)
 Lightning strike
 Bacillary haemoglobinuria
Black leg can be differentiated from edematic carbuncle form of
anthrax in that
 Anthrax affects all species of animals including equines.
 Edematic carbuncles because of black leg are crepitating but not in
anthrax
 Impression smears that are prepared from the ear of recently dead
animals from anthrax are capsulated with square ended bacilli but
not in black leg. 126
 Black leg cont’d---
Malignant edema occurs due to wounds.
In bacillary haemoglobinuria, there is a continuous urination of red
urine.
Treatment
It is possible to use antimicrobial drugs that have either narrow or
broad spectrum of action like
1. Penicillin groups like
Procaine penicillin in dose of 30,000 – 40, 000 IU/kg BW I/M
injection for 3 – 4 days or
Bezanthine penicillin in dose of 0.04 ml/ kgBW I/M injection for 3
– 4 continuous days.
2.Tetracycline groups like
 Oxytetracycline in dose of 2 – 10mg/ kg BW I/M injection for 3 –
4 days.
4. Aminoglycosides
Streptomycin sulphate in dose of 10 mg/KG BW I/m for 3 – 4
days. 127
 Black leg cont’d---
We can also use combinations drugs like penstrip and sulpha drugs.
It is also advisable to inject into the inflammatory edema either of
the following disinfectants like 3- 5% phenol or lysol; or 1 – 2%%
H2o2 or 0.1% kmno4.
Prevention and control measures
In farms where black leg is not registered prophylactic quarantine is
necessary.
While in area where black leg is enzootic ,annual vaccination of all
cattle at the age of 3 months and above; and sheep at 3 months age
and above must be practiced.
The most common black leg vaccine is formalin inactivated
aluminum hydroxide adjuvanated vaccine.
Vaccination should be done just prior to the anticipated danger
period, usually before pasture (before autumn and summer season in
Ethiopia case).
Dose cattle – 1ml/ SC in neck region
Sheep – 0.5 ml/ SC inner thigh of animal 128
 Black leg cont’d---
Antibody is produced after 10 – 14 days and lasts for 6 – 8 months.
NB. It is not advisable to vaccinate calves and lambs before 3
months of age respectively because maternal immunity persists
for at least 3 months in calves and for 6 months in lambs and will
interfere with active immunity.
In an outbreak areas all sick animals, animals those are suspecting
to be sick and suspected to be infested must be
 Isolated from the herd in separated isolation rooms
 Treated with appropriate antimicrobial drugs( sick and suspecting to
be sick)
 Isolation room and any object which has contact with sick animals
must be either burned or disinfected by appropriate disinfectants
like 4% formalin or 10% NaOH or sodium hypochlorite having 5%
active chlorine after mechanical removing of dirt. 129
 Black leg cont’d---
Dead animals must be either burned or buried.
Other animals in the herd should be vaccinated immediately
If antibiotics are not given, new cases of blackleg may occur
for up to 14 days until immunity develops, and constant
surveillance and the early treatment of cases will be necessary
Movement of the cattle from the affected pasture is advisable.
2.5.Malignant edema(clostridial myonecrosis)
It has also another name called gas gangrene or “false black
leg”.
Malignant edema is an acute, generally fatal toxemia affecting
all species and ages of animals characterized by myonecrosis
and gas gangrene.

130
 Black leg cont’d---
Species of Clostridium those cause malignant edema( gas gangrene)

Clostridium species Affected animals Diseases

C. septicum Cattle, sheep and pigs Malignant oedema
C. novyi (type A) Sheep Big head of ram
Cattle and sheep Gas gangrene
C. sordellii Cattle, sheep and horse Gas - gangrene

131
 Malignant cont’d---
Etiology
It is usually caused by histotoxic clostridia called Clostridium
septicum.
Other clostridial species have been isolated, indicating mixed
infections.
Additional clostridia implicated in wound infections include C.
chauvoei, C. perfringens, C.novyi and C. sordellii.
The bacteria are gram positive rod shaped with oval or spherical
shaped spore located sub terminal.
The bacteria form spores under aerobic conditions.
So that malignant edema is soil born disease.
They grow in anaerobic media like cooked meat broth with
thioglycollate(Robertson cooked meat medium) and on
conventional blood agar under anaerobiosis.
Epidemiology
The disease occurs worldwide.
All ages and species of animals are affected. 132
 Malignant cont’d---
The clostridia that cause malignant edema are common inhabitants
of the animal environment and intestinal tract and, although some
of the causative species have a restricted distribution.
The disease occurs sporadically, affecting individual animals
except in circumstances where a management procedure to a group
of animals results in an outbreak.
Source of infection
The infection is usually soil-borne.
The resistance of spores of the causative clostridia to
environmental influence leads to persistence of the infection for
long periods in a local area.
A dirty environment that permits contamination of wounds with
soil is the common predisposing cause.
Means of transmission
In most cases a wound is the portal of entry.
Deep puncture wounds accompanied by severe trauma provide the
133
 Malignant cont’d---
most favorable conditions for growth of anaerobes, and malignant
edema occurs most frequently under such conditions.
Deep puncture wound infection may occur through
Surgical or accidental wounds
Following vaccination and intramuscular injection of drugs
Venipuncture or through the umbilical cord in the newborn
After shearing and docking
Pathogenesis
C. septicum, C. novyi (type A) and C. sordellii are found in soil
and intestinal contents of animals throughout the world.
Infection ordinarily occurs through contamination of wounds
containing devitalized tissue, soil, or some other tissue debilitant or
through activation of dormant spores.
Wounds caused by accident, castration, docking, insanitary
vaccination, and parturition may become infected.
Potent neurotoxins are produced in the local lesion and cause death
when absorbed into the bloodstream. 134
 Malignant cont’d---
Locally the exotoxins cause extensive edema and necrosis followed
by gangrene.
Clinical findings
Clinical signs appear within 6 - 48 hours of infection.
The main clinical signs in malignant edema after predisposing
injury are:-
Anorexia
 High fever (41 - 42°C)
Weakness and show muscle tremor
Usually stiffness or lameness
 Local lesions, develop within 6–48 hrs at sites of injury
The local lesion at the site of infection consisting of a soft, doughy
swelling with marked local erythema accompanied by severe pain
on palpation.
At a later stage the swelling becomes tense and the skin dark and
taut.
The muscle in such areas is dark brown to black. 135
Malignant edema in calf and ram

136
Malignant edema in pigs

137
 Malignant cont’d---
Emphysema may or may not be present, depending on the type of
infection.
For example in C. novyi infections there is no emphysema.
The mucosae are dry and congested and have very poor capillary
refill.
Severe edema of the head of rams develops after infection of
wounds inflicted by fighting.
In “swelled head” of rams the edema is restricted initially to the
head.
It occurs first under the eyes and spreads to the subcutaneous
tissues of the head and down the neck.
When infection occurs at parturition, swelling of the vulva
accompanied by the discharge of a reddish-brown fluid occurs
within 2 - 3 days.
The swelling extends to involve the pelvic tissues and perineal
region.
138
Necropsy findings
Tissue changes occur rapidly after death, particularly in warm
weather, and this must be kept in mind when evaluating
postmortem findings.
The most common necropsy findings in malignant edema are:-
 Gangrene of the skin with edema of the subcutaneous and
intramuscular connective tissue around the site of infection.
The edema fluid varies from thin serum to a gelatinous deposit.
 It is usually bloodstained and contains bubbles of gas except in C.
novyi infections when the deposit is gelatinous, clear and contains
no gas.
The muscle in such areas is dark brown to black.
 Subserous hemorrhages and accumulations of serosanguineous
fluid in body cavities are usual.
In “swelled head” of rams, the necropsy findings is
 Edema of the head and neck that may extend into the pleural cavity
and also involve the lungs.
139
 Malignant cont’d---
Diagnosis
Diagnosis of malignant edema is done based on
Epidemiology of disease
Clinical signs
Necropsy finding
Laboratory
The first three methods of diagnosis give the tentative diagnosis of
the disease.
However, laboratory diagnosis is a confirmatory diagnosis
So the appropriate samples for confirmatory diagnosis are
Fascial tissue, placed in an airtight container
2 – 4 air-dried smears of fluid from lesion
Specimens must be taken as early as possible but not after 24 hours
of death.
Fascial tissues are used for bacterial isolation and identification by
culturing in anaerobic media or blood agar in anaerobiosis.
Air dried smears are used to identify the bacteria by serological 140
test
 Malignant cont’d---
called FAT(direct or indirect).
Tissues samples must be collected for histopathology by
preserving in 10 % formalin for histopathology.
Molecular techniques like PCR can be used for direct identification
and differentiation of clostridia associated with malignant edema.
Differential diagnosis
Malignant edema must be differentiated from
1. Blackleg
Black leg may occur with out external wound.
Horses and pigs are susceptible to malignant edema but not to
blackleg.
2. Braxy( bradsot) in sheep
C. septicum also causes braxy in sheep, a highly fatal infection
characterized by toxemia and inflammation of the abomasal wall.
This disease seems to be confined mostly to European sheep fed on
“frosted” pasture.
3. Carbuncle and angina form of anthrax 141
 Malignant cont’d---
No blacking of the muscles and subcutaneous tissues and no
emphysema.
4. Photosensitivity in white-faced sheep with swelled head
NB. The association of profound toxemia, local inflammation and
emphysema at the site of a wound is characteristics of malignant
edema to differentiate from the above diseases.
Diagnosis can be confirmed rapidly on the basis of fluorescent-
antibody staining of C. septicum from a tissue smear.
However, C. septicum is an extremely active postmortem invader
from the intestine, and its presence in a specimen taken from an
animal that has been dead for ≥24 hr is not significant.
Treatment
Malignant edema can be treated by using the following methods
Treatment of animals at early stage by using antitoxin has good
result but expensive.
Treatment with high doses of penicillin or broad-spectrum
antibiotics is indicated early in the disease. 142
 Malignant cont’d---
Although injection of penicillin directly into the periphery of the
lesion may minimize spread of the lesion.
Supportive therapy with NSAID (flunixin meglumine for cattle and
horses) is recommended.
Local treatment consists of surgical incision to provide drainage,
and irrigation with( 3 – 5%) hydrogen peroxide.
Control and prevention measures
Hygiene at lambing, shearing, castration and docking is essential to
the control the infection of sheep.
Vaccination with a specific or multivalent clostridial bacterins
toxoid prevents the occurrence of the disease in enzootic areas.
C. septicum usually is combined with C. chauvoei in a blackleg
vaccine and is available in multivalent vaccines.
Penicillin can be given prophylactically to animals at risk for the
disease.
In endemic areas, animals should be vaccinated before they are
143
castrated, dehorned, or docked.
 Malignant cont’d---
Calves should be vaccinated at the age of 2 months.
Two doses 2–3 wk apart generally give protection.
In high-risk areas, annual vaccination is indicated, as is
revaccination after severe trauma.
2.6. Infectious necrotic hepatitis (Black disease)
Infectious necrotic hepatitis(black disease) is an acute toxemia of
sheep; sometimes cattle and rarely in pigs and horses.
It is characterized by acute toxemia of sheep, cattle and sometimes
pigs and horses caused by the toxin of Clostridium novyi type B.
The exotoxin is elaborated from the bacteria that proliferate in
damaged liver tissue.
That is why black disease outbreaks are usually associated with
fasciolosis in sheep and cattle.
Since it causes congestion of venous blood vessels of the skin, in
some literature they call “black disease”.
144
Etiology
Black disease is caused by Clostridium novyi type B.
It is gram positive spore forming bacteria and resident in soil.
So that black disease is also soil born disease.
C. novyi type B are Gram positive rods that produces oval to
cylindrical sub terminal spores with little bulging of the mother
cell.
Their nutritional requirement differs from other clostridia in that
they require additional amino acid cysteine for their growth.
So that cooked meat medium(Robertson cooked meat medium) is
not used to isolate it instead anaerobic medium called Moore’s
medium( enriched with amino acid cysteine )is used.
Blood agar and/or stiff blood agar which are enriched with amino
acid cysteine can be used to isolate the pure colonies.
Not only the migrating metacercaria predispose the sheep and
cattle to black disease but also:-
Invasion of the liver by immature liver fluke
145
 Black disease cont’d---
 Local hepatic injury; e.g. invasion by cysts of Cysticercus
tenuicollis and other hepatic parasites
Trauma from liver biopsy
Epidemiology
Black disease affects mostly sheep and cattle.
The disease has worldwide in distribution but is of particular
importance in Australia and New Zealand.
The disease is becoming more common in some areas where liver
fluke is being introduced.
The disease is always fatal in both sheep and cattle.
Well-nourished adult sheep in the 2- 4years age group are
particularly susceptible.
Lambs and yearlings rarely being affected
Outbreaks are most common in the summer or autumn months.
This seasonal occurrence is marked because of fluctuation in the
liver fluke and host snail population.
Heavy irrigation of pastures creates favorable conditions for the 146
 Black disease cont’d---
development of flukes and may predispose disease.
Outbreaks in cattle and sheep commonly occur on irrigated farms
through out the year.
Source of infection
Fecal contamination of the pasture by carrier animals
Cadavers of sheep dead of the disease may cause heavy
contamination
Mechanism of transmission
The spread of infection from farm to farm occurs via carrier sheep
 Probably also by infected wild animals and birds
 By the carriage of contaminated soil during flooding
Pathogenesis
Spores of C. novyi type B are ingested and carried to the liver in the
lymphatic system.
The organism can be isolated from the livers of normal animals.
The spores may reach to liver and remain dormant in the kupffer
147
 Black disease cont’d---
cells.
Traumatic damage to the liver especially due to migrating
metacercaria and invasion of the liver by immature liver flukes
and other factors produce tissue damage and anaerobic condition .
In anaerobic environment the bacteria proliferate and liberating
alpha toxin( α – toxin) which is neurotoxin and causes local liver
necrosis and more diffuse damage to the vascular system and beta
toxin( β – toxin) which is phospholipase that acts on cell
membrane phospholipid bi – layer host cell.
The nervous signs observed may be due to this general vascular
disturbance or to a specific neurotoxin.
Clinical signs
Affected sheep ( usually 2 – 4 years old)commonly die during the
night with no well-defined signs and are found dead without
having exhibited any previous signs of illness.
The disease is most prevalent in well nourished adult sheep and
seems to be limited to animals infected with liver flukes 148
 Black disease cont’d---
Most cases occur in the summer.
Differentiation from acute liver flukes may be difficult.
But peracute deaths of animals that show typical lesions on
necropsy should arouse suspicion of infectious necrotic hepatitis.
Some common clinical signs in infectious necrotic hepatitis are:-
 Fever (40 - 42°C)
Rapid and shallow respiration
Sternal recumbency and often dies within a few minutes while still
in this position.
Clinical findings are the same in cattle as in sheep but the course is
longer, the illness lasting for 1-2 days.
Outstanding clinical findings in cattle include:-
 Sudden severe depression
Reluctance to move
Abdominal pain especially on deep palpation of the liver
Periorbital edema may also develop
149
 Black disease cont’d---
Necropsy findings
Bloodstained froth may exude from the nostrils.
The carcass undergoes rapid putrefaction.
There is pronounced subcutaneous edema.
Usually, there is extensive rupture of the capillaries in the
subcutaneous tissue, which causes the adjacent skin to turn black.
The dark appearance of the inside of the skin, particularly noticeable
on drying, has given rise to the name black disease.
Blood stained serous fluid is always present in large amounts in the
pericardial, pleural, and peritoneal cavities.
Subendocardial and subepicardial hemorrhages' are frequent.
Other common findings are an enlarged pericardial sac filled with
straw-colored fluid.
Excess fluid are found in the peritoneal and thoracic cavities.
However, the most characteristic gross lesions are seen in the liver
with characteristics of:-
Swollen liver 150
C. tenuicollis and F. hepatica which predispose animals to black
disease

151
The presence of yellow colored necrosis surrounded by a zone of
hyperemia in the liver

152
 Black disease cont’d---
 Grayish - yellow, necrotic foci in the liver along migratory tracks
of the young flukes.
 The necrotic foci are yellow areas 1 - 2 cm in diameter and are
surrounded by a zone of bright red hyperemia.
 These necrotic masses are found mostly under the capsule of liver
Diagnosis
Diagnosis of black disease is made by complex method including
Epidemiology
Clinical findings
Necropsy findings
Laboratory methods
In epidemiology consider:-
 Seasonal occurrence of the disease related to the migration of
immature liver fluke
Prevalent of the disease in young sheep with good body condition.
In clinical findings consider:-
Sudden death of animals 153
 Black disease cont’d---
The presence of pain around liver
Peritonitis
In necropsy findings consider:-
Rapid autolysis, engorgement of subcutaneous vessels with edema.
 Rupturing of subcutaneous capillaries that give the skin black
colour.
 The presence of yellow colored necrosis surrounded by a zone of
hyperemia in the liver
The above three diagnostic methods give tentative diagnose of the
disease.
Confirmatory diagnose is done by laboratory method.
Confirmatory diagnose of black disease requires:-
 Isolation and identification of C. novyi type B from the typical
liver lesion and
 Demonstration of preformed toxin in the peritoneal fluid and/or
the liver lesion from a fresh carcass.
Specimens of choice for isolation and identification of C. novi 154
 Black disease cont’d---
type B are liver and peritoneal fluids that are taken from recently
dead animals.
Pieces of liver tissue must be transported in air-tight container to
laboratory in cold chain for culturing in laboratory under
anaerobiosis.
2 – 4 impression smears from periphery of lesion( subcutaneous
tissues) must be prepared for FAT( direct and indirect).
 Additionally, liver tissue must be collected and preserved in 10 %
formalin for histopathology.
NB. Although Fluorescent Antibody Technique (FAT) is
almost as accurate and much less time consuming than traditional
anaerobic culturing methods, it has its own drawn back in
diagnosis of black disease due to the occurrence of nonpathogenic
strains of C. novyi type B.
These strains are detected in livers by the fluorescent antibody
technique and this may lead to a false-positive test result.
155
 Black disease cont’d---
An ELISA for beta toxin in intestinal contents has been described.
PCR techniques for better identification of toxin-producing
strains of clostridia are available.
Differential diagnosis
Black disease must be differentiated from:-
Acute fasciolosis in sheep
Other clostridial disease like blackleg and malignant edema
 Peracute form of anthrax
Treatment
No -effective treatment is available.
In cattle the longer course of the disease suggests the possibility of
controlling the clostridial infection by the parenteral use of
penicillin or broad-spectrum antibiotics.
However, the cases have high case fatality.
So that it is difficult to treat animals with black disease.
Prevention and control measures
The incidence of black disease may be lowered by 156
 Black disease cont’d---
Reducing the numbers of Lymnaea spp snails, the intermediate
hosts for the liver flukes.
 Reducing the fluke infestation of sheep
However, these procedures are not always practical and must be
used in combination with active immunization.
Active immunization of sheep is done with C.novyi type B
aluminum precipitated toxoid.
Active immunization can also be carried out not only as
prophylactic measures but also during outbreaks as ring
vaccination.
Long term immunity is produced by one vaccination( more than 4
years).
After this, only new introduced to the flock (lambs and sheep
brought in from other areas) need to be vaccinated.
This is best done in early summer.
Pasture contamination can be minimized by proper disposal of
157
cadavers (burning).
 Black disease cont’d---
2.7. Bacillary haemoglobinuria
The disease is also called “red water disease”.
Bacillary haemoglobinuria is an acute infectious, toxemic disease
of primary cattle but has also been found in sheep and rarely in
dogs and pigs caused by Clostridium novyi type D(Clostridium
haemolyticum).
Etiology
C. haemolyticum is a soil born organism naturally found in the GI
tract of some cattle.
It can survive for long periods in contaminated soil or in bones
from carcasses of infected animals.
It is gram positive spore forming bacteria and resident in soil.
So that bacillary haemoglobinuria is also soil born disease.
C. novyi type D(C. haemolyticum ) is gram positive rods that
produces oval to cylindrical, sub terminal spores with little bulging
of the mother cell.
158
 Bacillary cont’d---
Nutritional requirement differs from other clostridia in that they
require additional amino acid cysteine for growth as C. novyi type
B.
So that cooked meat medium(Robertson cooked meat medium) is
not used to isolate it; instead anaerobic medium called Moore’s
medium( enriched with amino acid cysteine )is used.
Blood agar and/or stiff blood agar which are enriched with amino
acid cysteine can be used to isolate the pure colonies.
Epidemiology
It occurs world wide including our country.
The cattle are the usual species involved although occasional cases
occur in sheep and rare cases in pigs.
As is the case in many clostridial diseases, animals in good
condition are more susceptible.
The disease is less common but mortality because of bacillary
haemoglobinuria may reach 25 % in infected farms.
Bacillary haemoglobinuria is a disease of the summer and autumn
159
 Bacillary cont’d---
months.
A primary association occurs with pastures that also are associated
with the occurrence of liver fluke although other, less determined
risk factors obtain.
The highest incidence of bacillary haemoglobinuria is on irrigated
or poorly drained pasture.
Some outbreaks have occurred in feedlots where hay cut from
infected fields was fed.
The disease is rare on dry, open range country but does occur in
range country where cattle have access to swales with areas
naturally irrigated by springs or streams.
Heavy mortalities may occur when cattle from an uninfected area
are brought on to an infected farm, cases beginning to occur 7-10
days later.
Source of infection
Fecal contamination of the pasture by carrier animals
160
 Bacillary cont’d---
Cadavers of sheep dead of the disease may cause heavy
contamination.
Mechanism of transmission
The disease is spread from infected to non infected areas by
Flooding and natural drainage
Contaminated hay from infected areas, or carrier animals.
 The carriage of bones or meat by dogs or other carnivores could
also effect spread of the infection.
Pathogenesis
Under natural conditions in endemic areas invasion occurs from
the alimentary tract after ingestion of contaminated materials with
spore of C. haemolyticum .
The spores, as in black disease of sheep may reach to liver and
remain dormant in the kupffer cells until favorable condition is
formed for their proliferation(tissue hypoxia).
Metacercaria and migrating young flukes are the primary factor
leading to liver necrosis and the establishment of anaerobic 161
 Bacillary cont’d---
conditions in the liver that will lead to the multiplication of the C.
haemolyticum.
Invasion of the liver by Cysticercus tenuicollis and other causes of
liver damage can also lead to the disease.
The development of an organized thrombus in a sub terminal branch
of the portal vein produces the large anemic infarct that is
characteristic of the disease.
Most of the bacteria are to be found in this infarct and, under the
anaerobic conditions.
Then the bacteria produce necrotic toxin and hemolytic beta toxin
and are released systemically and result in toxemia, generalized
vascular damage and intravascular hemolysis.

162
 Bacillary cont’d---
The major toxins produced by C. novyi types and diseases they
cause
C. novyi sub types Toxins

α - toxin β - toxin Disease

C. novyi type A + _ Big – head in young rams
Gas gangrene
C. novyi type B + + Black disease in sheep
C. novyi type C - _ Osteomyelitis in water buffalo
C. novyi type D - +++ Bacillary haemoglobinuria in
( C. haemolyticum) cattle occasional in sheep and
rarely in pigs

Systemic bacteriology and mycology

163
by Dereje Baye
 Bacillary cont’d---
Clinical signs
The illness is of short duration and cattle at pasture may be found
dead without signs having been observed.
If it has acute form , duration of clinical signs varies from almost
12 hrs in pregnant cows to 3–4 days in other cattle.
Usually, there is a sudden onset of severe depression
Fever that may reach 39. 5 – 41 0C
Abdominal pain and dyspnea
 The feces are dark brown; there may be diarrhea with much mucus
and some blood
The urine is dark red in colour(haemoglobinuria)
Anemia and jaundice are present in varying degrees
Grunting may be evident on walking
Respiration is shallow and labored and the pulse is weak and rapid
Pregnant cows often abort
Edema of the brisket is a common finding
The mortality in untreated animals is ∼95% 164
 Bacillary cont’d---
Some cattle suffer from subclinical attacks of the disease and
thereafter act as carriers.
Necropsy findings
After death, rigor mortis sets in quickly.
There is a variable degree of anaemia and jaundice.
Excessive amounts of fluid, varying from clear to bloodstained
turbid are present in the pleural, pericardial, and peritoneal
cavities.
The characteristic lesion of bacillary haemoglobinuria is an
ischemic infarct in the liver.
The infarct is pale, surrounded by a zone of hyperemia, and has the
general appearance of local necrosis.
The kidneys are dark, friable, and petechiation is evident
throughout the kidney.
The bladder contains purplish red urine.
Red urine is present in the kidneys and bladder
Diagnosis
Like other diseases diagnosis of bacillary haemoglobinuria can 165
be
Ischemic infarct of liver

166
 Bacillary cont’d---
done based on
 Epidemiology of the disease
 Clinical signs of the disease
 The necropsy finding
 Laboratory findings
The general clinical picture , clinical pathology and postmortem
findings usually permit a tentative diagnosis.
The most striking sign is the typical port-wine-colored urine,
which foams freely when voided or on agitation.
The clinical pathological features that permit a tentative diagnosis
are
The red color of the urine is due to the presence of hemoglobin
 Absence of free Red blood cells in the urine
 The presence of anemia followed by jaundice
 Reduction of erythrocyte count which is in between 1 and 4 x
109/ml
Reduction of hemoglobin concentration which is in between 0.003 167
 Bacillary cont’d---
Red urine due to the presence of haemoglubin in urine

168
 Bacillary cont’d---
0.008 gm/ ml
Leukocyte counts vary considerably from 6.7 - 3.48 x 109/ml.
Differential counts vary similarly, with a tendency to neutrophilia
in some cases.
Blood cultures during the acute stages of the disease may be
positive.
Serum agglutinins against C. haemolyticum may be detectable at
low levels (1:25 or 1:50) during the clinical illness.
 However, if the animal recovers, rise to appreciable levels (1:50
to 1:800) a week later.
But a positive agglutination test is not conclusive evidence of the
presence of the disease.
In necropsy findings, the presence of typical liver infarct is
sufficient for a presumptive diagnosis.
Diagnosis can be confirmed in laboratory by isolating C.
haemolyticum from the liver infarct.
Rapid and accurate diagnosis can be made by demonstrating the 169
 Bacillary cont’d---
organism in the liver tissue by
 Fluorescent antibody or
 Immuno histochemical test or
 Demonstrating the toxin in the fluid in the peritoneal cavity or in a
saline extract of the infarct in laboratory animals.
 Isolation identification of bacteria in Moor’s medium under
anaerobiosis
Differential diagnosis
The diagnosis of bacillary haemoglobinuria is largely a question of
differentiation from other diseases in which haemoglobinuria,
myoglobinuria, and haematuria are cardinal signs.
So that bacillary haemoglobinuria must differentiated from
Anthrax
Babesiosis and anaplasmosis
Enzootic haematuria
Hemolytic anemia caused by cruciferous plants
170
 Bacillary cont’d---
Bracken fern that cause enzootic haematuria

171
 Bacillary cont’d---
Cruciferous plants

172
 Bacillary cont’d---
Acute leptospirosis
Post parturient haemoglobinuria
Chronic copper poisoning (sheep)
Treatment
Treatment of bacillary haemoglobinuria comprises
Specific treatment
Supportive treatment
Specific treatment includes the immediate use of antibiotics like
penicillin or tetracyclines at high doses and antitoxic serum if
available.
Prompt(early) treatment is essential: provided that the serum is
administered in the early stages of the disease, haemoglobinuria
may disappear within 12 hours.
Supportive treatment including blood transfusion, parenteral
fluid and electrolyte solutions is of considerable importance.
Strong exercises may cause sudden death.
Bulls should not be used for service until at least 3 weeks after 173
 Bacillary cont’d---
recovery because of the danger of liver rupture.
Convalescence is often prolonged and animals should be protected
from nutritional and climatic stress until they are fully recovered.
Haemopoiesis should be facilitated by the provision of mineral
supplements containing iron, copper, and cobalt.
Prevention and control measures
Prevention measures of bacillary haemoglobinuria comprises
 Annual vaccination of animals with formalin-killed whole culture
adsorbed on aluminum hydroxide( used as adjuvant).
 Vaccination is carried out 4 - 6 weeks before the expected
occurrence of the disease.
 Annual revaccination of all animals over 6 months of age is
necessary in enzootic( endemic) areas.
 In some locations of extreme risk a second vaccination during the
grazing season is recommended.
 Cattle in contact with animals from areas where this disease is
enzootic( endemic) areas should be vaccinated, as the latter may 174
 Bacillary cont’d---
be carriers.
The carcasses of animals dying of the disease should be disposed of
by burning or deep burial.
2.8. Enteric diseases associated with Clostridium
perfringens
Clostridium perfringens resides in the intestinal tract of domestic
animals and can produce a number of toxins that result in enteric
and histotoxic diseases.
Clostridial enterotoxaemia are acute highly fatal intoxication that
affect sheep, lambs, calves, piglets and occasionally foals.
C. perfringens isolates are classified into one of the five types,
these are C. perfringens types A – E.
The diseases are caused by the major exotoxins(enterotoxins) of
Clostridium perfringens type B, C, D and occasionally type A and
E.
The major and minor toxins of C. perfringens play a very important
175
role in pathogenesis and pathogenicity of C.perfringens.
 Perfringens cont’d---
The minor toxins of Clostridium perfringens are:-
1. Theta (haemolysin)
 2. Kappa (collagenase)
 3. Mu (hyalurinidase)
 4. Nu (DNase)
They contribute the major tissue damage.
The major toxins of Clostridium perfringens are:-
1. Alpha
2. Beta
3. Epsilon
4. Iota
1. Alpha toxin
It is a lecithinase( phospholipase) toxin.
Attacks cell membranes causing cell death and destruction.
Alpha toxin is produced by all types of Clostridium perfringens(A
– E) and gives a zone of partial haemolysis on blood agar.
176
 Perfringens cont’d---
2. Beta toxin
It is lethal and necrotizing toxin produced only by Clostridium
perfringens type B and C.
Beta toxin is sensitive to trypsin and it is inactivated by trypsin.
That is why this toxin affects mostly neonates as colostrums has
antitrypsin activity.
Beta toxin is labile toxin and may be destroyed if there is
delayaness in submission of the specimen.
3. Epsilon toxin
Epsilon toxin is produced by C. perfringens type B and D .
Secreted by the bacterium as protoxin in intestine and activated by
protease such as trypsin.
This toxin causes pulpy kidney disease in sheep( C. perfringens type
D).
That is why pulpy kidney disease does not affect neonates.
As colostrum has antitrypsin factor.
No trypsin means no activation of the protoxin epsilon. 177
 Perfringens cont’d---
The pathogenesis of epsilon toxin is that it damages the epithelium
tissue of the gut and assuring absorption of the toxin into the blood
stream.
It damages vascular endothelium including blood vessels in the
brain leading to fluid loss and oedema.
That why the epsilon toxin can be regarded as an enterotoxin and a
neurotoxin.
4. Iota toxin
Iota toxin is produced by C. perfringens type E.
It is produced as protoxin but it is not fully under stood.

178
Pulpy kidney of sheep due to epsilon toxin of C. perfringens type D

179
 Perfringens cont’d---
The major toxins of Clostridium perfringens.
Types of
isolates Lethal toxins
Α( alpha) Β( beta) E( epsilon) I(iota)

A + - - -

B + + + -

C + + - -

D + - + -

E + - - +

180
 Perfringens cont’d---
The major diseases of animals caused by enterotoxaemic clostridia
C. Major Affected hosts Disease Clinical and post
perfringen toxins mortem findings
s type
A Alpha Lambs Enterotoxaemic Anemia, icterus,
jaundice haemoglobinuria and
death
B Beta, Lambs under 3 Lamb dysentery Hemorrhagic and rapidly
epsilon and weeks old fatal enterotoxaemia
alpha Lambs are often found
dead
C Beta and Pig lets 1 – 3 Hemorrhagic Dysentery, collapse and
alpha days old enterotoxaemia(clo death
stridial enteritis)

Lambs, foals Hemorrhagic Lumen is full of bloody

and calves enterotoxaemia( fluid
clostridial
enteritis)
181
 Table cont’d---
The major diseases of animals caused by enterotoxaemic
clostridia
C. Major Affected Disease Clinical and post
perfringens toxins hosts mortem findings
type
C Beta and Adult sheep Struck Sudden deaths due
alpha and goat to an enterotoxaemia

D Epsilon and Sheep all Pulpy kidney Oedema of brain,

alpha ages except disease glysosuria, sudden
neonates death.
Rare case in Excess fluid in body
calves and cavities.
goats Focal symmetrical
encephalomalacia in
some cases
E Iota and Calves and Enterotoxemia Pathogenicity
alpha lambs unclear.
182
 Perfringens cont’d---
Enterotoxemia caused by Clostridium perfringens types B and C
Disease C. perfringens type Host
Lamb dysentery Type B Lambs ≤3 wk old

Calf enterotoxemia Types B and C Well-fed calves ≤1 month

old
Pig enterotoxemia Type C Piglets in first few days
of life
Foal enterotoxemia Type B Foals in first week of life

Struck Type C Adult sheep

Goat enterotoxemia Type C Adult goats

183
 Perfringens cont’d---
Epidemiology
C. perfringens that cause different diseases in domestic animals are
found world wide.
The organisms are recoverable from the skin of sows, cows, mares
and ewes and in the feces of affected animals and infection of new
born animals probably occurs during suckling.
Insufficient cleaning and disinfection of the house of new born
animals with their mothers may cause to increase the incidence of
enterotoxaemia.
In lamb dysentery, the incidence of clinical disease in groups of
lambs may reach as high as 20 - 30% in intensive farms.
In an outbreak, the disease initially affects 1 - 4-day-old.
The case fatality rate approaches 100 % .
Type C enterotoxemia in lambs and goats occur as an outbreak with
an attack rate of 15 - 20% and a case fatality that approaches 100%.
184
 Perfringens cont’d---
Pathogenesis
The organism is ingested from soil and fecal contamination on the
surface of the dam's udder.
It proliferates and attaches to the surface of the epithelial cells of
the intestinal villi.
The factors that allow proliferation and attachment are poorly
understood.
Toxigenic strains of C. perfringens types B and C produce both
alpha and beta toxin.
The alpha toxin is a lethal toxin and is produced in varying
amounts by isolates of both types.
It is a phospholipase, and hydrolysis of membrane phospholipids
in erythrocytes, platelets, leukocytes, and endothelial cells results
in cell lysis or other forms of cytotoxicity.
The beta toxin causes increased capillary permeability and may
facilitate its uptake from the intestine.
Beta toxin is a necrotizing toxin and initially produces damage185to
 Perfringens cont’d---
the microvillus.
The age incidence of these diseases may be partially explained by
the observation of both beta and epsilon toxins.
Beta toxin is highly sensitive and inactivated by trypsin.
Colostrum contains a trypsin inhibitor that is why beta toxin of C.
perfergens type B and C mostly affect neonates.
While epsilon toxin of C. perfringens type B and D is produced in
protoxin form and it is activated by trypsin.
That is why pulpy kidney disease does not affect neonates which is
caused by epsilon toxin.
Clinical signs
Many of sick animals may die before clinical signs are seen.
However, some newborn lambs those do not die suddenly may
show the following clinical signs
 They stop nursing and become restless.
 They remain recumbent
 A fetid( unpleasant) blood-tinged diarrhea is common, and death186
 Perfringens cont’d---
usually returns within a few days.
In calves, there is
Acute diarrhea
Dysentery and abdominal pain
 Convulsions and opisthotonos
Death may occur in a few hours, but less severe cases survive for a
few days, and recovery is possible.
Pigs become acutely sick within a few days of birth and there is
Diarrhea
Dysentery
Reddening of the anus
High fatality rate; most affected piglets die within 12 hrs
In foals, there is
Acute dysentery
Toxemia and rapid death
Struck in adult sheep is characterized by death without premonitory
signs. 187
 Perfringens cont’d---
Enterotoxaemic disease caused by C. perfringens type D called
pulpy kidney disease has the following clinical signs :-
Sudden deaths in the best-conditioned lambs are the first indication
of enterotoxemia.
In some cases excitement, incoordination, and convulsions occur
before death.
Opisthotonos, circling, and pushing the head against fixed objects
are common neurologic signs
Hyperglycemia or glycosuria is present
Diarrhea may or may not develop.
Occasionally, adult sheep are affected too, showing weakness,
incoordination, convulsions, and death within 24 hr.
In goats, the course of disease ranges from peracute to chronic,
with signs that vary from watery diarrhea with or without blood to
sudden death.
Affected calves not found dead show mania, convulsions, blindness,
188
and death within a few hours.
 Perfringens cont’d---
NB. The important clinical pathology of enterotoxemia caused by
C. perfringens type D is glycosuria.
Necropsy findings
Opening of the carcass of animals died because of enterotoxemia is
possible only for diagnostic purpose in special slaughter houses.
The major necropsy findings in young lambs during enterotoxaemia
caused by C. perfringens type A, B, Care
Hemorrhagic enteritis with ulceration of the mucosa of duodenum
Grossly, the affected portion of the intestine has deep blue-purple
colour
Pericardial sac is filled by fluid
Mesenteric lymph nodes are enlarged, hyperemic with foci of
necrosis.
There is large amount of serous – hemorrhagic exudate in thoracic
and abdominal cavities
Edema and malacia( softness) can be detected microscopically in the
basal ganglia and cerebellum of lambs. 189
 Perfringens cont’d---
In older sheep
Hemorrhagic areas on the myocardium
Petechiae and ecchymoses of the abdominal muscles and serosal of
the intestine.
 Frequently, there is bilateral pulmonary edema and congestion of
lung
The rumen and abomasum contain an abundance of feed, and
undigested feed often is found in the ileum.
Hemorrhagic or necrotic enterocolitis may be seen in goats.
Rapid postmortem autolysis of the kidneys has led to the popular
name of pulpy kidney disease.
Diagnosis
Diagnose of enterotoxemia can be done based on
Epidemiology of the disease
Clinical signs
Necropsy findings
190
 Perfringens cont’d---
Laboratory
A presumptive diagnosis of enterotoxemia is based on
 Sudden and convulsive deaths in lambs and typical necropsy
findings.
However, the confirmatory diagnosis of enterotoxemia is done
By extraction of toxins in intestinal contents followed by its
identification by using toxin protection( neutralization) test first
with poly- valent, followed bi - valent antitoxin then with mono
valent anti toxin in laboratory animals(mice or guinea pig).
So the appropriate samples are
For bacteriology
20 - 30 ml of intestinal content, frozen in a glass or plastic leak
proof container ( for anaerobic culturing, bioassay of toxins and for
PCR)
 Air-dried smears of mucosal surface from several levels of small
intestine (Cyto - Gram stain)
 Collect tissue samples of several segments of each for histology 191
 Perfringens cont’d---
Toxin identification method by toxin neutralization in bioassay
Toxins of Clostridium perfringens types A to E

Antitoxin A B C D E
Alpha toxin Alpha Alpha Alpha Alpha
Beta Beta Epsilon Iota
Epsilon
Type A: p d d d d
Anti - alpha
Type B: p p p p d
Anti alpha
Beta
Epsilon
Type C: p d p d p
Anti -
alpha
beta
192
 Table cont’d---
Toxin identification method by toxin neutralization in bioassay
Toxins of Clostridium perfringens types A to E
Antitoxin A B C D E
Alpha toxin Alpha Alpha Alpha Alpha
Beta Beta Epsilon Iota
Epsilon
Type D: p d d p p
Anti –
alpha
epsilon
Type E: p d d d d
Anti alpha
Iota
NB. “P” means protected no death
“ d” means not protected, dead

193
Hemorrhagic enteritis because C. perfringens type A, B, C

194
Enterotoxaemia ( Dilated intestine showing a patchy congestion)

195
 Perfringens cont’d---
pathology in 10 % of formalin
Although immunologic tests have been developed to replace the
traditional mouse assay for detection of toxin, they are less
sensitive.
A PCR for detection of epsilon toxin gene is available for
identification of the isolates as either type B or D.
Differential diagnosis
Enterotoxemia must be differentiated from many animal diseases
like
Bradsot( braxy)
 Peracute and acute form of anthrax
Septicemic form of pasteurellosis
Listeriosis
Salmonellosis
Enteric colibacillosis
Cryptosporidiosis
Coccidiosis 196
 Perfringens cont’d---
Strongyloides westeri
NB. The most important way to differentiate them is
bacteriological, parasitological and serological laboratory methods.
Treatment
In individual cases the disease is often too acute for effective
therapy.
However, if you have got time use
1. Specific therapy by using
Specific hyperimmune serum
Antimicrobials per os and parenteral
Oral and parenteral administration of penicillin or tetracycline etc
may prevent further proliferation of organisms and production of
toxins
2. Supportive therapy 197
 Perfringens cont’d---
Administration of fluid and solutions to control dehydration and
acidosis etc.
3. Symptomatically therapy
Remove undesired clinical signs by administration of different
drugs.
Prevention and control measures
The method of prevention measures depends on
The age of the lambs
The frequency with which the disease appears on a particular
property
 The method of husbandry
If the disease is seen consistently in young lambs, ewe
immunization probably is the most satisfactory method of control.
Breeding ewes should be given 2 injections of type D toxoid in their
first year, a booster injection 4 – 6 weeks before lambing, and each
year thereafter.
198
Enterotoxemia in feedlot lambs can be controlled by immunization
 Perfringens cont’d---
Enterotoxemia in feedlot lambs can be controlled by immunization
of all lambs with toxoid upon entering the feedlot.
Two injections with 2 weeks apart, will protect lambs through the
feeding period.
However, when an outbreak occurs
Antitoxin can be administered to all sheep as soon as an outbreak
commences because
It can be used to treatment infected animals in combination with
antimicrobials
It can be also used to develop passive immunity for non – infested
animals.
If enterotoxaemia is endemic in the area
Susceptible pregnant animals should be vaccinated either by
bacterins or toxoid to provide colostral immunity to their progeny.
Vaccination, preferably with type-specific toxoid or bacterins, is
the specific preventive measure.
If there is an outbreak of enterotoxaemia, toxoid is cheaper, but to
199
 Perfringens cont’d---
administer it at such times may result in further serious losses
before active immunity develops.
So inactive out breaks it is better to use specific antitoxin than
toxoid.
NB. You can administer antitoxin in dose of 150 – 200 IU/ kg BW
IV and provide protection almost for 1 month.
The carcasses of animals dying of the disease should be disposed of
by burning or deep burial.
2.1. Ulcerative lymphangitis
The other name of the disease is pseudotuberculosis.
It is acute, subacute, chronic and latent bacterial infection of many
domestic animals characterized by the formation of suppurations or
granuloma(casein nodes) in different organs of affected animals.
The granulomas that are formed in different organs resemble the
granuloma of tuberculosis.
That is why the disease and the positive agent are named
200
2. Chronic bacterial diseases of large animals
In this portion we give attention to:-
2.1. Ulcerative lymphangitis
2.2. Tuberculosis
2.3. Paratuberculosis
2.4. Brucellosis

201
2.1. Ulcerative lymphangitis
The other name of the disease is pseudotuberculosis.
It is acute, subacute, chronic and latent bacterial infection of many
domestic animals characterized by the formation of suppurations
or granuloma(casein nodes) in different organs of affected animals.
The granulomas that are formed in different organs resemble the
granuloma of tuberculosis.
That is why the disease and the positive agent are named
pseudotuberculosis and Corynebacterium pseudotuberculosis
respectively.
In horses the disease is called ulcerative lymphangitis.
Ulcerative lymphangitis in horses is an infection of the lower
limbs, abscesses in the pectoral region and ventral abdomen and
abscess in different internal organs.
In cattle, the bacteria most commonly cause cutaneous excoriated
granulomas.
Location on the animal is variable but it is often associated with
202
skin trauma.
 Ulcerative cont’d---
Abortion and mastitis may also occur in pregnant mares and cows.
Rarely, visceral involvement has been reported in cattle.
In small ruminants C. pseudotuberculosis cause caseous
lymphadenitis (CL).
Caseous lymphadenitis (CL) is a chronic, contagious disease caused
by the bacterium Corynebacterium pseudotuberculosis.
Caseous lymphadenitis is characterized by:-
 Abscess formation in or near major superficial lymph nodes
(external form) or
 Abscess formation in the thorax and abdomen (internal form).
The internal form of caseous lymphadenitis can cause ill thrift and
respiratory compromise and is a major rule out for the “thin ewe”
syndrome.
Caseous lymphadenitis is primarily a disease of sheep and goats.
But sporadically it can also affect horses, cattle, swine and
camelids.
Caseous lymphadenitis occasionally can affect people. 203
 Ulcerative cont’d---
So appropriate precautions should be taken when handling
infected animals and purulent exudate from lesions.
Etiology
Ulcerative lymphangitis and caseous lymphadenitis are caused by
C. pseudotuberculosis.
The bacteria have straight to slightly curved pleomorphic rods,
frequent club-shaped swellings.
C. pseudotuberculosis is a gram-positive, facultative intracellular
coccobacilli.
It can occur in rod, coccoid club and filamentous shapes.
Stained smear from animal tissue often reveal as group of cells.
These cells may present in parallel(palasides) or cells at sharp
angles to each other( Chinese letters).
Corynebacteria are non-spore forming, non motile and non- acid
fast bacteria.
Many have metachromatic granules(high energy phosphate stores).
204
 Ulcerative cont’d---
Because it exhibits the property of metachromasia, where in the
granules appear in a colour other than the colour used for staining.
The granules that give such property are made up of
polymetaphosphates.
Two biotypes have been identified based on the ability bacteria to
reduce nitrate: -
 A nitrate-negative group that infects sheep and goats, and
 A nitrate-positive group that infects horses.
 Isolates from cattle are a heterogeneous group.

205
C. pseudotuberculosis

206
 Ulcerative cont’d---
Epidemiology
In natural condition the diseases caused by C. pseudotuberculosis
affects small ruminants, horses, cattle, pigs and camel.
Ulcerative lymphangitis mostly has sporadically character.
But rarely it can occurs as epidemics(epizootics) in winter season.
The bacteria can be found in the soil, water and in animal feeds.
The main reservoir of C. pseudotuberculosis are rodents and mice.
Source of infection
They are sick and recovered animals that distribute the organism
together with:-
Pus from infected superficial lymph nodes
Pus discharges through nostrils

207
 Ulcerative cont’d---
Mechanism of transmission
Ingestion
Aerosol
Cutaneously through damaged skin( hair shearing, castration, biting,
umbilicus and arthropods as mechanical vectors like stable flies,
horn flies)
Most infections occur through contact with purulent exudate from
ruptured external or pulmonary abscesses.
Skin injuries from shearing, tagging, docking, castration,
environmental hazards (eg, splintered wood, metal edges, jutting
nails, wire) provide opportunity for establishment of infection.
Use of communal sheep dips can spread the disease, because C.
pseudotuberculosis can survive in dip solutions for up to 24 hr.
So that shearing immediately before dipping increases the risk of
infection.
C. pseudotuberculosis does not multiply in the environment, but it
can survive on hay, straw, and wood for 2 months, and in soil for2088
 Ulcerative cont’d---
months.
The bacteria enter via skin wounds by arthropod vectors such as
stable flies, horn flies and house flies or by contact with
contaminated fomites or soil.
Pathogenesis
Corynebacteria are pyogenic bacteria causing varieties of
suppurative conditions.
All strains produce an exotoxin called phospholipase D.
Phospholipase D enhances dissemination of the bacteria by
damaging endothelial cells and increasing vascular permeability.
The bacterium has a second virulence factor, an external lipid coat
that provides protection from hydrolytic enzymes in host
phagocytes.
Replication of bacteria occurs in the phagocytes, which then
rupture and release bacteria.
The ongoing process of bacterial replication, followed by attraction
209
and subsequent death of inflammatory cells, forms the
 Ulcerative cont’d---
characteristic abscesses associated with caseous lymphadenitis .
Infection occurs after C. pseudotuberculosis penetrates the skin or
mucous membranes.
Clinical signs
The onset of ulcerative lymphangitis in horses is variable and may
manifest :-
 As painful swelling, pustules, and ulcers, especially in the region
of the lower limbs
 Lameness and edematous swelling extending up the entire limb.
 The exudate is odorless, thick, tan and blood tinged and usually,
only one leg is involved.
Ulcerative lymphangitis in cattle is characterized by:-
 Diffuse or localized swellings, ventral pitting edema, ventral
midline dermatitis, lameness and draining abscesses.
 Abscessation of the pectoral region or ventral abdominal region
with secondary dissemination to internal organs.
 A marked or prolonged fever, anorexia, or weight loss indicates 210
 Ulcerative cont’d---
toward sequelae such as deep or recurring abscesses, internal
abscessation, or systemic infection with abortion.
Abscesses can be large, upto 20 cm in diameter before rupturing,
and take weeks to months to resolve.
Superficial abscesses eventually rupture and discharge infectious
purulent material into the environment
The skin wound heals and leaves a scar.
The infection can spread in blood or lymph to internal lymph nodes
and to visceral organs such as the lungs, kidney, liver, uterus and
brain.
Less common sites of involvement include the udder, scrotum, and
joints.
You know the diseases caused by C. pseudotuberculosis in small
ruminants is called caseous lymphadenitis.
However, the distribution of abscesses in sheep and goats differ
somewhat possibly as a result of management differences.
External abscesses around the head and neck occur more commonly211
 Ulcerative cont’d---
in goats.
However, the visceral form is more common in sheep.
Bovine ulcerative lymphangitis

212
 Ulcerative cont’d---
Ulcerative lymphangitis in equine

213
 Ulcerative cont’d---
Ulcerative lymphangitis in equines

214
 Ulcerative cont’d---
Caseous lymphadenitis in sheep and goats

215
 Ulcerative cont’d---
Caseous lymphadenitis in sheep and goats

216
 Ulcerative cont’d---
Necropsy findings
Very often the carcass of died animals are emaciated.
Superficial cranial lymph nodes including the visceral lymph nodes
of lung usually enlarged and have casein.
The casein is covered by strong capsule made of connective tissue(
except in goats)
The casein mass has layered structure while you cut it(Except in
goats).
In visceral form of ulcerative lymphangitis you may observe small
grayish, gray – green colour nodules in lungs, liver, spleen and
kidneys that have casein.
In horses you always find lymphangitis while in small ruminants
not lymphangitis but lymphadenitis.
In sheep, the abscess often has the classically described laminated
“onion-ring” appearance in cross section with concentric fibrous
layers separated by inspissated caseous exudate.
217
 Ulcerative cont’d---
In goats, the abscesses are less organized, and the exudate is usually
soft and pasty.
Onion-ring” appearance of lymph node of goat affected by C.
pseudotuberculosis in cross section

218
 Ulcerative cont’d---
Diagnosis
Since the clinical forms of equine and bovine ulcerative
lymphangitis may appear in different forms, it has a little value to
diagnose the disease.
However necropsy findings help us to reach the tentative diagnose
of the disease.
Confirmatory diagnose is done in bacteriological laboratory.
Isolation and identification of C. pseudotuberculosis from lesions is
necessary for confirmation.
In all forms of lymphangitis in horses and cattle, samples for culture
include
Aspirates of abscesses
 Swabs of purulent exudate beneath crusts associated with folliculitis
A punch biopsies
The presence of an external abscess on a small ruminant is highly
suggestive of caseous lymphadenitis especially in an endemic herd.
But an aspirate from an intact abscess should be submitted for 219
 Ulcerative cont’d---
bacteriologic culture for definitive diagnosis.
Animals with visceral abscesses pose a greater diagnostic challenge
So, several attempts are made to diagnose the visceral form of
ulcerative lymphangitis(UL) and caseous lymphadenitis(CL.
Among them the most common are:-
Radiography and ultrasonography
 Culture of a transtracheal aspirate obtained from an animal with
pneumonia can help determine if caseous lymphadenitis is the
cause.
Serologic testing
Abdominal radiography and ultrasonography is useful for
detection of internal infection of the liver, spleen, or kidneys or
under muscle fascias.
Transtracheal aspirates are required to confirm pneumonia caused
by C. pseudotuberculosis.
Serologic testing with the synergistic hemolysis inhibition test,
which detects IgG to the phospholipase D exotoxin, is a useful 220
 Ulcerative cont’d---
adjunct for diagnosis of internal infection
This test detects antibodies to the phospholipase D exotoxin.
Titer results must be evaluated in light of the herd history, presence
or absence of clinical disease, and history of caseous lymphadenitis
vaccination.
In none vaccinated herd, titers of 1:8 and higher are considered
indicative of infection and titers of 1:256 and higher are correlated
with internal abscessation.
However, synergistic hemolysis inhibition serological test has its
own drawn back because
 The test never differentiate a vaccinated animal from a naturally
infected animal.
 It never detect the early infested animals and false - negative
results can occur if testing is done in the first 2 weeks after
exposure before the animal has seroconverted.
 Also, animals with chronic walled-off abscesses can have adjunct
for diagnosis of internal infection and show false-negative result.
221
 Ulcerative cont’d---
NB. When the status of an animal with a positive titer is in doubt,
the titer should be repeated after 2–4 weeks.
Differential diagnosis
Ulcerative lymphangitis in horses and in cattle, and caseous
lymphadenitis in small ruminants must be differentiated from
Tuberculosis
Other bacterial species that can cause suppuration and
lymphangitis(Staphylococcus aureus, Rhodococcus equi,
Streptococcus spp, Actinomycetes spp, Actinobacillus spp)
Strangles
Glanders
Dermatophilosis
Epizootic lymphangitis
Dermatophytosis
Sporotrichosis
 Equine cryptococcosis
222
 Ulcerative cont’d---
Blastomycosis
Parasitic disease onchocerciasis
Treatment
Treatment of ulcerative and caseous lymphangitis is effective if the
lesions are superficial.
If there is pain full external swellings, treated with hot packs of
water( hydrotherapy) or Ecitamol ointment for abscess maturation.
Then the matured abscess ruptures if not drain the abscess
surgically and lanced and flushed with dilute antiseptic solutions
like iodine solution, cresol and lysol solution.
The purulent material should be collected into a disposable
container and incinerated.
The surgeon should wear disposable gloves to avoid inadvertent
self-inoculation.
Treated animal should be isolated from other animals until the
wound is healed.
Deep abscesses under muscles like triceps or quadriceps region is 223
 Ulcerative cont’d---
difficult to diagnose and to treat.
That is why internal infection carries a 30 – 40% mortality rate,
even with appropriate treatment.
NB. Severe or untreated lymphangitis cases often become chronic,
and fibrosis and induration of the leg occurs.
Prevention and control measures
If the farm is free from the disease, all new animals and feed of the
animals must be from UL and CL free areas.
Quarantine for at least for 30 days is necessary for replacement
animals.
Attention must be given to zoohygienic condition of the farm by
doing prophylactic disinfection, deratization and disinsection.
Producers should remove hazardous items (barbed wire, exposed
nails, rough feeders) from the environment to decrease injury and
potential CL transmission.
Producers should also purchase their own shearing equipment and
dip solutions, and not share it with other flocks. 224
 Ulcerative cont’d---
Older animals and those with abscesses should be shorn last, and
require ultrasonography to guide placement of an indwelling drain.
Deep abscess drainage require to administer anti inflammatory and
pain relievers like phenylbutazone.
General supportive and nursing care is indicated.
Lymphangitis and internal infection should be treated with long term
antimicrobials (1 month duration or as directed by follow-up
ultrasonography).
The efficacy of systemic antimicrobial therapy is controversial.
Although C. pseudotuberculosis is susceptible to many antibiotics
“in vitro” ,treatment is not necessarily effective “in vivo “.
Because the antibiotic cannot penetrate into the abscesses well.
So that it is better to use antibiotics that have ability to penetrate the
abscesses well like rifampin or rifampin in combination with other
antibiotics.
Long term (4 – 6 weeks) penicillin (22,000 IU/kg, IM) and rifampin
(10 – 20 mg/kg, PO ) has been used to treat the internal caseous 225
 Ulcerative cont’d---
form of lymphadenitis with limited success.
Equipment should be disinfected after contamination by exudate.
Shearers should arrive at the flock with clean hands and wearing
clean footwear and clothing.
All items that come in contact with animals from different flocks
should be sterilized, replaced, or thoroughly cleaned between flocks
But if there are cases of UL and CL in your farm
Isolate sick animals and treat them.
Vaccinate the rest of the herd by CL vaccine( it is available in
Europe and USA).
Vaccination of young replacement stock reduces the incidence and
prevalence of CL within a flock.
Currently, all contain phospholipase D toxoid, and some also
contain killed whole bacterial cells.
The vaccine is available as a monovalent bacterin and as a
polyvalent preparation combined with Clostridium tetani and C.
226
perfringens type D.
 Ulcerative cont’d---
The initial dose is given S/C in the axillary space after colostral
immunity has waned (~3 months of age) and should be repeated in
4 weeks.
Colostral immunity can be improved by administering a booster to
pregnant ewes and does 1 month before lambing/kidding.
Lambs and kids from infected dams can be raised on pasteurized
colostrum and milk away from infected animals.
Vaccination of the young replacement stock should be considered,
and older infected animals should be gradually culled as economics
allow.
Once the disease is at a low prevalence rate, vaccination should be
stopped and all seropositive unvaccinated animals culled.
Replacement animals should be purchased from producers with
good CL prevention programs whenever possible.
Only seronegative animals, and those with no evidence or scars or
abscesses near lymph nodes, should be allowed into the clean herd
227
or flock.
 Ulcerative cont’d---
NB. The presence of infected asymptomatic animals in the “clean”
herd or flock limits the success of this approach, and is one reason
why eradication of CL is difficult.
2.2. Tuberculosis
Tuberculosis (TB) is an infectious chronic granulomatous disease of
many domestic and wild animals characterized by the formation of
granulomatous tubercles in different organs.
The disease affects practically all species of vertebrates and, before
control measures were adopted, it was a major disease of humans
and domestic animals.
Bovine TB is still a significant zoonosis in many parts of the world
specially in developing countries.
The current increasing incidence of tuberculosis in humans,
particularly in immuno - compromised humans, has given a
renewed interest in the zoonotic importance of M. bovis, especially
in developing countries
228
 TB Cont’d---
M. bovis can be responsible for 5 to 10% of human tuberculosis with
higher rates in children in some areas.
Infection in humans occurs largely through:-
 Consumption of infected raw milk and raw milk products by
children
Inhalation
So that transmission of bovine tuberculosis to humans can be
significantly reduced by pasteurization of milk.
But only complete eradication of the disease can protect the farmer
and his family.
It is under strict control in most developed countries but is still a
major cause of economic loss in many less well endowed countries.
Economic loss due to bovine tuberculosis comprises from:-
Actual deaths
 Lose of 10 - 25% productive efficiency of affected animals
 Lose because of discarding and condemnation of slaughtered
animals’ carcass organs with tuberculous lesions and the entire 229
 TB cont’d---
carcass or
The requirement of heat treatment before being released for human
consumption.
Etiology
The microorganism is found under genus Mycobacterium.
There are different species of Mycobacterium that affect domestic
animals.
Both of them have similar morphology, staining and cultural
characteristics.
All are acid – fast, aerobic, non -motile, non spore forming and
non - capsulated bacteria.
They are cultured in egg yolk based artificial medium called
Lowenstein – Jensen medium with malachite green or crystal
violet inhibitors.
The pathogenic species of Mycobacterium are slow growing
bacteria in the laboratory.
Because most of them require more than 3 weeks for growth 230
Acid fast staining when methylene blue is used as a counter staining
dye

231
Acid fast staining when malachite green is used as counter staining

232
Lowenstein – Jensen medium with malachite green and crystal
violet inhibitors

233
Lowenstein – Jensen medium with crystal violet inhibitors

234
 TB cont’d---
(except M. avium that requires 2 – 6 weeks).
However there are many species of Mycobacterium that can grow
within either <7 days or >7 days but not > 14 days.
These species of Mycobacterium are called non- pathogenic or
atypical mycobacterium.
So that it is easy to differentiate the pathogenic species of
Mycobacterium from atypical ones by their time requirement for
growth in laboratory.
But there are many pathogenic species of Mycobacterium that has
not been cultured yet "in vitro”.
These are:-
 M. lepraemurium
 M. leprae
 The non – identified acid fast mycobacteria which are causing
bovine skin tuberculosis.
Since we are studying large animals medicine, we are going to
focus only species of Mycobacterium that affect large animals. 235
 TB cont’d---
These are:-
M. bovis
M. avium
The rest are studied under human and small animal medicine .

236
Species of Mycobacterium that cause tuberculosis in human being
and animals
Species of Affected animals and Disease
Mycobacterium human
M. africanum Human Human tuberculosis (Africa)
M. tuberculosis Humans, dogs, Human tuberculosis( world wide)
canaries and psittacine
birds
M. bovis Many animal species Bovine tuberculosis
and humans
M. avium Poultry and wild birds Avian tuberculosis
Generalized form rare in mammals
Pigs Lesions in cervical lymph nodes and
intestinal lesions
Horses Rarely intestinal lesions
M. microti Voles Vole tuberculosis.
Localised lesions in rabbit, calves
and guinea pigs. 237
Epidemiology
Large number of species of domestic and wild animals are
susceptible to M. bovis.
All species, including humans, and age groups are susceptible to M.
bovis.
Cattle, goats, pigs and dogs are the most susceptible species.
While sheep, horses and cat showing a high natural resistance.
From wild animals, monkeys are the most suceptible animals to M.
bovis.
Source of infection
Infected animals( cattle) are the main source of infection for other
cattle.
Organisms are excreted together with:-
 Exhaled air and in sputum from pulmonary lesions
 Feces (from both intestinal lesions and swallowed sputum from
pulmonary lesions)
Milk
238
 TB cont’d---
Sperm, urine, vaginal and uterine discharges
Discharges from open peripheral lymph nodes
In general animals with gross lesions that communicate with
airways, skin, or intestinal lumen are obvious disseminators of
infection.
Infested animals including cattle in the early stages of the disease,
before any lesions are visible, may also excrete viable
mycobacteria in nasal and tracheal mucus.
In epidemiology of bovine tuberculosis wildlife reservoirs play an
important rule.
Because while most wildlife are unimportant as sources for
infection to cattle, in some areas of the world certain wildlife
species appear to be a significant maintenance host.
These wild life can transmit bovine tuberculosis to our domestic
animals by:-
Contamination of pastures by their urine
239
 TB cont’d---
Visiting farm buildings and cattle troughs to feed during which
they defecate and urinate directly onto the cattle feed.
 Direct contact on the pasture-bush margin where there is ample
opportunity for cattle-possum contact.
NB. Many wild animals are maintenance host for bovine
tuberculosis.
For example many studies show that buffalo is the maintenance
host for bovine tuberculosis in South Africa.
Means of transmission
There are different ways of transmission.
These can be grouped as:-
Ingestion
Inhalation
Other routes
Ingestion and inhalation are the common entry of M. bovis.
However, ingestion the most common entry of the organism.
For example 240
 TB cont’d---
 Pigs can be infected by M. bovis or M. avium when they eat raw
infected internal organs of cattle and other animals.
Hens are infected by ingestion of infected feed.
 Young animals can be infected through milk from infected animals.
Infection by ingestion is possible at pasture when feces contaminate
the feed and communal drinking water and feed troughs but a large
infective dose is required.
So that environmental contamination of pasture is not of major
importance in the epidemiology of the disease in cattle.
This is because the organism can survive for long periods in feces
and soil but most studies show that survival on pasture is measured
in weeks rather than months.
Inhalation is almost invariable portal of entry in intensive farms
where sick, latent carriers and healthy animals are kept in bad
ventilated animal houses.
Other route of transmission includes
 Placental transmission of fetus 241
 TB cont’d---
 Transovarian transmission in birds
Intrauterine infection at coitus( natural insemination),
Intrauterine infection by the use of infected semen or of infected
insemination gun or uterine pipettes
Intramammary infection by the use of contaminated teat siphons or
by way of infected cups of milking machines.
Unusual sources of infection are infected cats, goats, or even
humans.
 For example stockmen with genito-urinary infections have
transmitted infection to cattle through urinating in the cattle
environment.
Animals can be infected by M. tuberculosis by direct contact with
infected human beings.
There are many risk factors for the occurs of tuberculosis in
animals.
Among the most importants are:-
Environmental 242
 TB cont’d---
Host risk factors
 Pathogen risk factor
Housing predisposes the animals to the disease, as does high
stocking density.
The closer the animals are in contact the greater is the chance that
the disease will be transmitted.
In addition to the above, bad microclimatic condition ( coldness,
poor in ventilation) aggravates the problem.
That is why tuberculosis nowadays one of the disease of
intensification.
Zebu (Bos indicus) type cattle are thought to be much more
resistant to tuberculosis than European cattle.
All species, including humans, and age groups are susceptible to M.
bovis.
Cattle, goats, pigs and dogs are the most susceptible species.
While sheep, horses and cat showing a high natural resistance.
From wild animals, monkeys are the most suceptible animals to243
 TB cont’d---
tuberculosis
The causative organism is moderately resistant to heat, desiccation,
and many disinfectants.
It is readily destroyed by direct sunlight unless it is in a moist
environment.
In warm, moist, protected positions, it may remain viable for weeks.
Pathogenesis
Inhalation of infected droplets expelled from the lungs is the usual
route of infection
Ingestion, particularly via contaminated milk, also occurs.
Intrauterine and coital methods of infection are recognized less
commonly.
Inhaled bacilli are phagocytosed by alveolar macrophages that may
either clear the infection or allow the mycobacteria to proliferate.
Phagocytosed M. bovis may pass to the regional lymph nodes and
finally to thoracic lymph ducts with general dissemination.
Phagocytosed M. bovis at the site of contact with alveolar
macrophages form primary focus.
Primary focus formation is mediated by cytokines associated with 244
a
 TB cont’d---
hypersensitivity reaction that consists of dead and degenerate
macrophages surrounded by epithelioid cells, granulocytes,
lymphocytes, and later, multinucleated giant cells.
The purulent to caseous, necrotic center may calcify, and the lesion
may become surrounded by granulation tissue and a fibrous capsule
to form the classic “tubercle.”
The primary focus plus similar lesions formed in the regional lymph
node is known as the “primary complex.”
At the center of the lesion caseous necrosis develops.
Caseous necrosis may proceed to calcification (cattle) or
liquefaction.
Once cell – mediated immunity is established, the spread of the
bacteria through lymphatic system is retarded.
However the bacteria may spread by contiguous extension or via
erosion of bronchi through blood vessels to a new area.
Haematogenous dissemination may produce miliary tuberculosis in
245
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animals.
This involves multifocal tubercle formation in an organs like liver,
kidney, skeleton, mammary glands, reproductive tract, and CNS or
in the serosal surface of a cavity like pleura and peritoneum.
In alimentary forms of disease, the primary focus may be found in
the pharynx or mesenteric lymph nodes or, less commonly, in the
tonsils or intestines.
The cellular composition of and presence acid-fast bacilli in
tuberculous lesions differ between and within host species.
The primary complex seldom heals in animals and may progress
slowly or rapidly.
Dissemination through vascular and lymphatic channels may be
generalized and rapidly fatal, as in acute miliary TB.
A prolonged, chronic course may also ensue, with lesions usually
having a more localized pattern of distribution.
246
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Clinical signs
Tuberculosis usually has chronic form with incubation period of 15
– 45 days.
At the beginning of infection, it is difficult to differentiate infected
animals from healthy ones.
The clinical signs reflect the extent and location of lesions.
However, the generalized clinical signs include
Progressive emaciation and lethargy
Weakness and anorexia
Fluctuating fever
Forms of tuberculosis in animals
There are different forms of tuberculosis that can be classified by
different characteristics.
1. By location of pathological process
Tuberculosis can be:
 A. Localized tuberculosis
 B. Generalized tuberculosis 247
 TB cont’d---
A. Localized tuberculosis
It is tuberculosis that affect only certain organs of animals and does
not have a tendency to disseminate.
It can be:
Pneumonic tuberculosis
Intestinal tuberculosis
Genital tuberculosis
Serosal membrane of cavity( pleura and peritoneum) tuberculosis
Tuberculosis of udder , kidney, liver, spleen, bone, bone marrow
and skin etc
B. Generalized form of tuberculosis
This type of tuberculosis is common in animals with low resistance.
Because the fibrous capsule of tubercle may breaks down and the
bacteria may travel through lymph and blood to other organs which
results formation of a lot of tubercle in different organs called
miliary tuberculosis.
248
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2. Relation of affected organs to the external environment
Conditionally it is classified as
A. Open tuberculosis
B. Closed tuberculosis
A. Open tuberculosis
It is also called active tuberculosis when the bacteria are expelled to
the external environment from affected organ together with milk,
faces , discharges, air and sputum.
So that mostly intestinal, udder, reproductive organs tuberculosis
and non encapsulated form of pneumonic tuberculosis are called
open type tuberculosis.
B. Closed tuberculosis
It is a type of tuberculosis characterized by absence of expelling of
the organism to the environment from the affected organs.
This is because the tubercle is encapsulated by fibrous connective
tissue and may be up to calcification.

249
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Forms of tuberculosis in different species of animals
1. Cattle
Cattle has the following forms of tuberculosis
Pneumonic
Intestinal
Serosal
Generalized ( rarely)
Reproductive organs
Mastitic
1. Pulmonary involvement is characterized by a chronic cough due
to bronchopneumonia.
The cough is never loud or paroxysmal occurring only once or twice
at a time and is low, suppressed, and moist.
It is easily stimulated by squeezing the pharynx or by exercise and
is most common in the morning or in cold weather.
A chronic, intermittent, moist cough with later signs of dyspnea
250
and tachypnea and with time coughing becomes painful.
 TB cont’d---
The destructive lesions of the granulomatous bronchopneumonia
may be detected on auscultation and percussion.
Dullness on percussion are accompanied by areas in which
squeaky crackles are audible by auscultation.
Very often they are audible over the caudal lobes of lung.
In this form of tuberculosis, there may be involvement of lymph
nodes.
Involvement of the bronchial lymph nodes may cause dyspnea
because of constriction of air passages.
While enlargement of the mediastinal lymph nodes is commonly
associated with recurrent and then persistent ruminal tympany.
The animal shows inappetance and reduction of productivity with
anemic visible mucous membrane.
2. In intestinal form of tuberculosis in cattle there is
 Progressive diarrhea results in emaciation
 Enlargement of retropharyngeal lymph nodes results in dysphagia
and noisy breathing due to pharyngeal obstruction. 251
 TB cont’d---
 Chronic, painless swelling of the submaxillary, prescapular,
precrural and supramammary lymph nodes is relatively rare.
3. Tuberculous pleurisy may occur but is usually symptomless
because there is no effusion.
4. In generalized form of tuberculosis in cattle
There is extensive miliary tubercular lesions but the animal looks
clinically normal.
 However, in most cases there is progressive emaciation
unassociated with other signs occurs, and should arouse suspicion of
tuberculosis.
 A capricious appetite and fluctuating temperature are also
commonly associated with the disease.
The hair coat may be rough or sleek.
Affected animals tend to become more docile and sluggish.
Superficial lymph nodes(submadibular, prescapular, precrural and
supramammary lymph nodes) are enlarged and less moveable.
252
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5. Tuberculosis affecting reproductive organs of cattle
This form of tuberculosis is not common in cattle.
However, rarely it affects uterus of cows and testicles of bulls.
Infection of uterus by tuberculosis cause metritis.
Tuberculous infection from uterus can spread by contiguity and
causes
Peritonitis
Bursitis
Salpingitis
The lesions in the salpinx taking the form of small enlargements
containing a few drops of yellow fluid.
In tuberculous metritis, there may be
Conception may be followed by recurrent abortion late in pregnancy,
or
Alive calf is produced which in most cases dies quickly of
generalized tuberculosis.
253
Infertility
 TB cont’d---
Lesions similar to those of brucellosis occur on the placenta.
In cows that fail to conceive, there maybe a chronic purulent
discharge
 Number of cows will have an associated tuberculous vaginitis
affecting chiefly the ducts of Gartner
 Rare cases of tuberculous orchitis are characterized by the
development of large, indurated, painless testicles
Lastly, affected animals become infertile
6. Tuberculous mastitis
It has major importance because of
 The danger to public health
Spread of the disease to calves
 Difficulty of differentiating it from other forms of mastitis
In this form of mastitis, there is
Enlargement of supramammary lymph nodes that become hard,
rough and less moveable.
254
A marked induration and hypertrophy which usually develops first
 TB cont’d---
in the upper part of the udder( in rare quarters).
As the result ,there is disconfiguration of udder.
Affected quarter is painless in palpation.
In the early stages, the milk is normal macroscopically
 However, later the milk becomes watery mixed with blood or
caseous mass.
2. Tuberculosis in pigs
Tuberculosis in pigs has usually latent form.
But very often tuberculous lesions in pigs are found in cervical
lymph nodes.
Mostly the lesions are located in submadibular and retropharyngeal
lymph nodes.
No clinical abnormality is observed on them unless they rupture to
the exterior.
However, caseous mass mixed with pus is expelled during surgical
opening of the abscess.
In generalized cases a syndrome similar in cattle can be seen. 255
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3.Tuberculosis in horses
The commonest syndrome in horses is caused by involvement of the
cervical vertebrae in which a painful osteomyelitis.
Less common signs include
 Coughing due to pulmonary lesions
 Lymph node enlargement
Nasal discharge and fluctuating temperature
4. Tuberculosis in small ruminants
Bronchopneumonia is the commonest form of the disease in these
species.
In some goats intestinal ulceration, with diarrhea, and enlargement
of the lymph nodes of the alimentary tract occur.
Necropsy findings
Macroscopic lesions of bovine tuberculosis are typically
characterized by the presence of tubercles of different sizes with
central caseation and calcification.
In the early stages of infection, these lesions are not encapsulated,
256
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but are surrounded by condensed alveolar tissue.
After long time of infection, tubercle may calcify.
Bovine tuberculosis mostly affects lung tissue and lymph nodes of
thoracic cavity, and forms tubercle of different size that is solid and
has casein at the center.
Large numbers of tubercle are found on pleura and peritoneum
when the organism affects the serose membranes of the two
cavities.
Tubercle lesion on different organs of cattle
Lung

257
 TB cont’d---

Lymph nodes

Miliary tubercle on spleen

258
 TB cont’d---
Tubercle lesion on lung and lymph node

259
 TB cont’d---
Lung parenchyma is almost entirely replaced by variably-sized,
coalescing, raised pale nodules.

260
 TB cont’d---
Bovine, uterus the endometrium contains numerous raised
tubercles.

261
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Pig, tracheobronchial lymph nodes. The center of the sectioned
node is replaced by caseous, mineralized debris.

262
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Miliary tubercle on mesenteric lymph nodes of intestine
Tubercle lesion on mucous membrane of
uterus

263
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Mycobacterium bovis granulomas medial surface of rib cage, cow

264
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Miliary tubercle on peritoneum and pleura

265
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Intestinal form of bovine tuberculosis is characterized by the
formation of ulcers on the mucous membrane of intestine.
In histopathological examination, initially, there are epithelioid and
giant cells at the center of the tubercle.
As the disease progress, they are surrounded by lymphocytes,
plasma cells and monocytes, developing a peripheral fibroplasias
and central caseous necrosis.
Necropsy finding in pigs are mostly found in mesenteric lymph
nodes and in lymph nodes of head and rarely in liver.
In poultry necropsy findings of tuberculosis infection are found
mostly in liver, spleen, bone and bone marrow.
Diagnosis
Diagnose of tuberculosis is done by complex methods based on
Epidemiology of the disease
Clinical signs
Allergic tests
266
 TB cont’d---
Serological and blood testing method
Necropsy findings
Bacteriological method
Molecular method
Diagnosis based on clinical signs alone is very difficult, even in
advanced cases
This is because
The clinical signs of tuberculosis are not typical
Absence of clinical signs in early infection
The most important diagnostic methods of tuberculosis on live
animals are
Allergic
Serological and blood testing methods
Radiography (nonhuman primates and small animals)
Microscopic examination of sputum and other discharges is
sometimes used not widely used in veterinary,
However, the most important and practically applicable diagnostic 267
 TB cont’d---
methods of tuberculosis on live animals are
Allergic method
Serological and blood testing methods
Allergic method
The delayed-type hypersensitivity response of the host, responsible
for much of the pathology of TB.
It is fundamental to the tuberculin skin test that is widely used for
diagnosis in large animals( cattle, equines, sheep, goat, pig and
poultry)
Delayed hypersensitivity reaction is used in live animals to know
whether the animal is infected with tubercle bacilli or not as the
disease is chronic.
Delayed hypersensitivity test is screening test
It is diagnosed by intradermal inoculation of tuberculin.
Screening testing of our herd for tuberculosis by intradermal
injection of tuberculin is called tuberculinization.
268
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Tuberculin is a Purified Protein Derivatives ( PPD) which is
prepared from M. bovis or M. avium.
PPD is a complex mixture of protein, lipid, carbohydrate and
nucleic acids derived from M. bovis or M. avium.
There fore, there are two types of PPD
PPD derived from M. bovis
PPD derived from M. avium
PPD of M. bovis is used to diagnose tuberculosis in mammals by
using allergic test.
While PPD of M. avium is used to diagnose tuberculosis in pigs and
poultry by using allergic test.
Intradermal skin testing using PPD has proven an effective
diagnostic test used for screening
M. bovis infected cattle and other domestic animals
M. avium infected poultry and pigs
Tuberculin test is also used for screening test to diagnose
tuberculosis in poultry and pigs. 269
 TB cont’d---
Techniques and procedures
There are two methods of tuberculinization techniques
1. Single intradermal tuberculinization
2. Comparative simultaneously intradermal tuberculinization
1. Single intradermal cervical tuberculinization
In this method M. bovis tuberculin is injected intradermal either in
the skin of caudal fold or in the middle third of the side of the neck
or lips of vulva.
Procedure
 1.The animal is identified (by its ear tag) and its identification is
recorded.
2. Injection site is selected in the middle third of the side of the neck
or caudal fold.
3. Hair is clipped around the site to a radius of about 2 centimeters.
4. A fold of skin is measured with calipers and the measurement is
recorded.
5. Tuberculin is injected into the skin at the dose of 0.2 ml. 270
 TB cont’d---
Single intradermal cervical tuberculinization
1. Measuring of the thickness of the skin
at the site of injection with caliper

2. Clipping of the hair

271
 TB cont’d---
Tuberculin, disposable and automatic tuberculin syringes

272
 TB cont’d---
Intradermal injection of tuberculin

Measuring of the thickness of the injection site of the skin after 72

hours with caliper

273
 TB cont’d---
Intradermal injection of tuberculin

Tuberculin test in caudal

skin folds

274
 TB cont’d---
Measuring of the thickness of the injection site of the skin after
72 hours with caliper

275
Tuberculin skin test in pigs

276
 TB cont’d---
 6. After 72 hours, the tester returns, confirms the animal identity,
measures the fold of skin and records the thickness of the skin fold.
If the animal is infected by M. bovis ,there will be swelling
(thickening of the skin) at the site of injection due to delayed type
hypersensitivity reaction( mononuclear cell infiltration which cause
oedema).
 NB. SID test has its own drawback because there is a cross
reaction with other species of Mycobacterium and non -
Mycobacterium species that can give false positive test result.
Among species of Mycobacterium and non – mycobacterium
species that can give false positive test
results are
 1. Non – identified acid fast bacteria causing bovine skin
tuberculosis
2. M. avium
3. M. tuberculosis
4. Saprophytic mycobacteria 277
 TB cont’d---
 5. M. paratuberculosis
6. Nocardia asteroids( none Mycobacterium species)
That is why in areas with a high incidence of either avian TB,
atypical mycobacteriosis, or paratuberculosis, comparative
simultaneously intradermal tuberculinization test is used.
2. Comparative simultaneously intradermal tuberculinization
It is a comparative simultaneously intradermal injection of PPD
(tuberculin of both M. bovis and M. avium) which can be
inoculated simultaneously but at separate sites in the neck.
Procedure
1.The animal is identified (by its ear tag) and its identification is
recorded.
2. Two injection sites are selected in the middle third of the side of
the neck, one above the other, separated by about 130mm (if it is a
small animal, the two sites will be on either side of the neck).
3. Hair is clipped around the sites to a radius of about 2 cm.
4. A fold of skin at both sites is measured with calipers and the 278
 TB cont’d---
measurements must be recorded.
5. Tuberculin is injected into the skin; the upper site is used for the
avian tuberculin (or use the left side of neck for small calves).
6. After 72 hours, the tester must return and identify the animal
identity and measure the thickness of skin at both sites and records
the thickness of the skin fold.
Interpreting the results
Usually, a “standard interpretation "of the skin test results is
applied.
If the reaction to bovine tuberculin is by 3mm(more) greater than
the reaction to avian tuberculin; the animal is considered to be
infected with M. bovis and is called a “reactor”.
If the reaction to bovine tuberculin is by 1 up to 3 mm greater than
the reaction to avian tuberculin, the animal is considered an
“inconclusive reactor "and will be retested after 60 days.
If the reaction to bovine tuberculin is by 1 mm and less as compare
the reaction to avian tuberculin, the animal is considered negative
279
to M. bovis infection.
 TB cont’d---
NB. There is also false negative test result because of anergy.
Anergy is the unresponsiveness state of organism to allergen.
The reason of anergy is not clear up to now but it may be due to poor
immunity such as
Those in the early stages of infection
Loss of body condition
Young animals(calves up to six months of age)
Anergic cases in advanced disease or old animals
Cattle that have recently calved
Pregnant animals also may have false negative results
Developed countries have regular testing policy for bovine
tuberculosis in every 6 months.
What about our country? As tuberculosis is the major public health
problem and dairy production in on way of intensification.
NB. Tuberculin test can be performed for all susceptible animals
including human beings.
However, in addition to cattle; pigs and chicken are animals on 280
Comparative simultaneously intradermal tuberculinization

281
Tuberculinization in different species of animals
Species of Site of tuberculin Dose of tuberculin Evaluation of
animals injection test reaction
Bovine Neck region or caudal 0.2 ml After 72 hours
fold
Sheep and Inner thigh of hind legs 0.2 ml After 48 hours
goat around the skin of femoral
bone
Pigs External surface of ear, 2 0.2 ml After 48 hours
cm at margin
Dogs Inner thigh of hind legs 0.2 ml After 48 hours
around skin of femoral
bone
Camel Skin of abdominal cavity 0.2 ml After 72 hours
specially on crural region
at the level of tuber ischii
Poultry Wattle 0.1 ml After 36 hours

282
 TB cont’d---
which tuberculinization is commonly performed.
There is also ophthalmic method of tuberculinization which is
mostly used for race horses.
It is done by two times dropping of 4 drops(for each) PPD into the
conjunctiva of the lower eyelid with 5 -6 days interval.
The test result is evaluated after every 6, 9, 12 and 24 hours of the
first; and after every 3, 6, 9 and 12 hours of the second test.
The test result is consider as positive when there is secretion of
mucosal – pus or pus secretion which is accompanied by hyperemia
and edema of conjunctiva.
Serological tests
Serological tests have been developed to diagnose bovine
tuberculosis includes
Complement fixation Test(CFT)
 Fluorescent Antibody Test(FAT),
Direct bacterial agglutination and precipitation tests
 Passive haemagglutination tests(indirect haemagglutination) 283
 TB cont’d---
But have little potential value for the routine diagnosis of
tuberculosis.
Even enzyme-linked immunosorbent assay (ELISA) tests to crude
mycobacterial antigens had limited value.
However, an ELISA which examines antibody to defined antigens
of M. bovis before and after skin testing appears useful in detecting
non -specific reactors.
An “in vitro” assay of cell-mediated reactivity by detection and
quantitation of “ γ”interferon known as the interferon- γ assay is
licensed and commercially and available in some countries.
It is based on the detection of IFN - γ liberated from white blood
cells in whole blood(lymphocytes) cultures incubated with PPD
tuberculin.
It is done by culturing and incubation of lymphocytes with
specific antigen (PPD - tuberculin) for 16 – 24 hour.
The test makes use of the comparison of IFN-γ production
following stimulation with avian and bovine PPD. 284
 TB cont’d---
The detection of bovine IFN-γ is carried out with a sandwich ELISA
that uses two monoclonal antibodies to bovine gamma-interferon.
Gamma interferon test has an advantage over other tests in that
It can detect infected cattle earlier in infection than can the skin
test.
 It also has value in retesting skin-test positive cattle that may be
false-positive reactors.
Necropsy finding
Necropsy findings of the classic “tuberculous” granulomas are often
very suggestive of the disease.
Bacteriological method
This is a confirmatory diagnose to isolate and identify species of
bacteria.
The specimens for isolation and identification of Mycobacterium
species can be
 1. From live animal or
 2. From dead animals 285
 TB cont’d---
1. Specimens from live animals
It includes
Aspirates from cavities
Lymph nodes biopsies
Tracheobronchial lavages
 The centrifuged deposit of 50 ml milk in the case of suspected
tuberculous mastitis
2. Specimens from dead animals
Specimens from dead animals include
 Fresh tissues with typical tubercle lesions in cold conditions(
collect from the same tissue for histopathology).
 If there is no lesion, take samples from selected lymph nodes from
tuberculin reactors.
Specimens except milk must be decontaminated, digested and
centrifuged before they are inoculated to Lowenstein – Jensen
medium.
But if the specimen is milk, no need of decontamination and 286
Procedure for recovering of Mycobacterium spp from specimens

287
 TB cont’d---
digestion as the it is comparatively sterile.
Take 50 ml of milk and centrifuge it.
Isolation is made by culturing specimens in selective medium for
mycobacteria called Lowenstein – Jensen (LJ) medium.
Identification can be done by
Growth time and colony morphology (eugonic = M. bovis and M.
avium and dysgonic = M. bovis)
By biochemical tests like requirement of glycerol for growth;
enhancement of growth with 0.4 % sodium pyruvate; niacin
production, pyrazinamidase; urease test and nitrate reduction test etc
Bioassay method by using rabbit, guinea pigs and chickens as
shown below.
Molecular method of diagnosis
There are many molecular techniques that have been developed for
direct detection of mycobacteria from clinical samples and from
288
Culture of M. bovis

289
Bioassay method of identification

290
 TB cont’d---
pure isolates.
They are based on amplification of unique mycobacterial DNA or
RNA target fragments by PCR.
Tests can be done on sputum, blood, nasal swabs, and other tissues,
with the advantage of rapidly detecting nonviable bacilli.
Polymerase chain reaction technique for identification of M. bovis
from tissues, milk and colonies, demonstrating that this test can be
valuable for rapidly identifying M. bovis in milk.
Differential diagnosis
Bovine tuberculosis must be differentiated from many subacute and
chronic infectious diseases of animals that have similar clinical
signs and necropsy findings.
So that it must be differentiated from
Contagious Bovine Pleuro Pneumonia (CBPP)
291
 TB cont’d---
Pneumonic form of pasteurellosis
Non – infectious bronchopneumonia
Aspiration pneumonia (which is often secondary to chronic wasting
disease in cervids)
Traumatic reticulopericarditis
Caseous lymphadenitis in small ruminants
Melioidosis in small ruminants and chronic aberrant liver fluke
infestation
Lymphadenopathy
Other causes of mastitis (to differentiate from tuberculosis mastitis)
Treatment
Treatment has been attempted using drugs that have had success in
humans, eg, isoniazid(isonicotinylhydrazin),ethambutol and
rifampin, rifampicin and pyrazinamide,
But in veterinary, it is not advisable to treat tuberculosis
Because
 It requires long term treatment, so it is not economically visible 292
 TB cont’d---
 Sick animals can be source of infection to other animals and human
beings
 Danger of encouraging drug resistance
Prevention and control measures
In tuberculosis free farms, prevention measures must be directed
 Keeping of the farm from other tuberculosis non – free farms.
 Periodically( at least every 6 months) to do tuberculin test to isolate
diseased animals on time.
Prophylaxis disinfection of the farm with appropriate disinfectants
But if tuberculosis is common in the farm the following eradication
programs are applied in different countries
 Test-and-slaughter or
 Test-and-segregation methods
Affected herds are re-tested periodically every 6 months to
eliminate cattle that may shed the organism.
293
Tuberculin test is generally used.
 TB cont’d---
Those that are positive reactors are considered infected herds and
are usually isolated and slaughtered.
Animals that have been in contact with reactors are traced by
tuberculin test.
Only test-and-slaughter techniques are guaranteed to eradicate
tuberculosis from domesticated animals.
However, some countries use test-and-segregation programs during
the early stages of eradication, and switch to test-and-slaughter
methods in the final stage.
Sanitation and disinfection may reduce the spread of the agent
within the herd.
Sanitation and disinfection may reduce the spread of the agent
within the herd.
Mycobacterium is relatively resistant to many disinfectants and
requires long contact times for inactivation.
Effective disinfectants are 5% phenol, iodine solutions with a high
294
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concentration of available iodine, glutaraldehyde and formaldehyde
Treatment is illegal in some countries.
The BCG (Bacillie Calmette-Guérin) vaccine, sometimes used to
control TB in humans, has proved to provide little protection in
most animal species but it is not widely used.
5. John's disease (Paratuberculosis)
Paratuberculosis is a chronic, contagious granulomatous enteritis
characterized in cattle by persistent diarrhea, progressive weight
loss, debilitation, and eventually death.
The disease is called John’s disease and the positive agent was
called M. johne.
Because in 1895, German physician called Heinrich Albert Johne
and his American colleague, Frothingham, were the first to
describe the disease.
But nowadays it is called bovine paratuberculosis. 295
 Para cont’d---
Etiology
The etiologic agent, Mycobacterium paratuberculosis, also known
as Mycobacterium avium subspp. paratuberculosis.
Mycobacterium paratuberculosis is thin, rod shaped, acid fast
bacteria.
It is aerobic, non – capsulated and non- spore forming bacteria.
It is slow growing bacteria and require 5 – 14 weeks for growth.
Like other species of Mycobacterium, M. paratuberculosis never
grow on/ in conventional media.
So that Herrold’s egg yolk medium with mycobactin is often used as
a medium to isolate the bacteria.
In specific broth media, M. paratuberculosis produce chemical
substance called paratuberculin or johnin which is essential for
allergic test.
Epidemiology
Paratuberculosis very often affects cattle and sheep.
Rarely buffalo, camel and very rarely goat. 296
 Para cont’d---
The disease occurs worldwide most commonly in cattle and to a
lesser extent in sheep and goats.
Very often the disease appears sporadically and very often affects
cattle up to 4 months age.
Predisposing factor of paratuberculosis are
Unbalanced ration (mostly Ca and P)
Bad conditions in housing and management
There are many risk factors for occurrence of paratuberculosis.
They are
 Age of animals: - Calves are more susceptible than other animals.
 Breed : -Predominant breed in the area is more susceptible.
Stress factors: - like parturition, transportation and nutritional
deficiencies and animal diseases like bovine viral diarrhea(B
VD)can aggravate paratuberculosis
 Herd characteristics: - like herd size, annual herd birth rate,.
annual herd replacement rate etc have influence on the occurrence
of paratuberculosis. 297
 Para cont’d---
Source of infection
The source of infection are
Sick animals
Carrier animal with latent form of infection
 M.paratuberculosis is excreted in large numbers in feces of infected
animals and in lower numbers in their colostrum, milk and uterine
discharges.
Means of transmission
Contaminated water, feed and fomites.
Susceptible animals can be infected by
Ingestion(fecal-oral route); calves can also be infected with
colostrum or milk
 Vertical transmission of paratuberculosis(transplacental
transmission)
Spread of the organism from farm to farm is usually due to trading
298
 Para cont’d---
of livestock which are unknown infected carriers and shedders of
the organism but lateral spread of feces across boundary fences
also occurs.
There are also many insects and internal parasites that plays a very
important rule in transmission.
These are
 Cockroach(serve as a passive vector)
Earthworms and adult Diptera
 Trichostrongylid larvae (Haemonchus contortus, Ostertagia
circumcincta, Trichostongylus colubriformis).
Pathogenesis
After ingestion and uptake in the Peyer's patches of the lower
small intestine.
This intracellular pathogen infects macrophages of the GI tract
and in associated lymph nodes.
299
 Para cont’d---
It is possible that some animals may eliminate infection through a
cell-mediated immune response that encourages bactericidal activity
in macrophages.
However in most cases of paratuberculosis, there is interaction of
bacteria and macrophages that cause inflammation.
This leads the mucous membrane of the intestine to be thickened
and corrugated because of cellular infiltration.
The organisms multiply and eventually provoke a chronic
granulomatous enteritis that interferes with nutrient uptake and
processing, leading to the cachexia in typical advanced infections.
Profuse diarrhea is not necessarily be seen in sheep and goats but
emaciation is caused by malabsorption of protein intestine.
Animals are thought to become infected in neonatal period .
Not all infected animals become clinical cases but they remain
subclinical excretors.
Clinical signs
Incubation period of the disease is 1 – 1 2 months and more. 300
 Para cont’d---
Paratuberculosis always has chronic form.
It has 3 stages
Latent
Subclinical
Clinical
Latent stage of paratuberculosis is characteri z ed by absence of
typical clinical signs.
However, infected animals have low growth rate and low body
condition.
It is possible to know whether the animals are infected or not by
allergic, serological and bacteriological methods of diagnosis.
In sub clinical stage some clinical signs may appear which are not
clear and typical for the disease.
The clinical stage of the disease in cattle is characteri z ed by
301
Para cont’d---

302
 Para cont’d---
 Weight loss and diarrhea in the late phases of infection
 Constant or intermittent diarrhea which does not contain blood
mucus, or epithelial debris and is passed without tenesmus.
Fading of coat colour and
 Edema around ventral and interamandibullar region due to a
protein-losing enteropathies.
This leads to low concentrations of total protein and albumin in
plasma.
Animals are alert, and temperature and appetite are usually normal,
although thirst may be increased.
The disease is progressive and ultimately terminates in emaciation
and death.
In infected herds, the mortality rate may be low for a number of
years
But up to 5 0 % of animals may be infected subclinically with
associated production losses.
303
 Para cont’d---
In sheep, goats and other ruminants, diarrhea may not be seen
Necropsy finding
Carcasses may be emaciated with loss of pericardial and perirenal
fat in the more advanced, cachectic cases.
A diverse array of pathology may be seen in infected animals,
ranging from a complete lack of gross lesions to a thickened and
corrugated intestine with enlarged and edematous neighboring
lymph nodes.
Intestinal lesions can be mild, but typically the distal small-
intestinal wall is diffusely thickened
L esions may extend proximally and distally to the jejunum and
colon.
Sheep, goats, and deer sometimes develop foci of caseation with
calcification in the intestinal wall and lymph nodes.

304
 Para cont’d---
Thickened and corrugated intestinal mucosa due Johne's disease

305
 Para cont’d---
Thickened and corrugated intestinal mucosa due to
paratuberculosis

306
 Para cont’d---
Thicken wall of small intestine due to paratuberculosis

307
Diagnosis
Diagnosis of paratuberculosis is done based on
Epidemiological
Clinical signs
Necropsy and histopathological findings
Allergic
Laboratory
Allergic test
Field diagnostic test is based on delayed hyper sensitivity reaction
to antigen called johnin.
Johnin is an antigen prepared from M. paratuberculosis used to test
paratuberculosis in animals.
It can be injected either intradermal or intravenous.
Intra dermal test is done by injecting johnin intradermal and
swelling of skin at site of injection after 48 – 72 hours considered as
positive.
It is practically not applicable because 70% of the test result is false
positive. 308
 Para cont’d---
Intravenous injection of johnin can be done by injecting johnin
intravenously.
The elevation of the body temperature of animals is considered as
positive.
It is useful only for clinical sick animals from paratuberculosis and
detect only 80% of animals.
It is not useful for asymptomatically shedders.
So it is not also practically applicable.
Laboratory diagnose
Laboratory diagnose comprises
Bacteriology
Serology
Molecular techniques
Specimens for bacterial isolation and identification can be
Small punch of biopsy from rectum or rectal scrapings
Faecal samples can be used but it is less important than the above
samples. 309
 Para cont’d---
 Ileocaecal valve which is free from faces and mesenteric lymph
nodes are specimens from dead animals.
Additional section of ileocaecal valve must be preserved in 10 %
formalin for histopathology.
In general the specimen of choice for isolation and identification of
M. paratuberculosis are mesenteric and ileocaecal lymph nodes,
ileum and liver.
The bacteria is isolated by Herrold’s egg yolk medium with
mycobactin.
It needs 7 – 15 weeks for growth and only mycobactin dependent
bacteria that is M. paratuberculosis grows there.
Identification can be done by
Microscopic appearance by Ziehl-Neelsen(ZN) staining.
Test on laboratory animals
 DNA probes
Serology
Serologic tests are rapid, low-cost methods for antemortem 310
Herrold’s egg yolk medium with mycobactin

311
ZN stained tissues from ileocaecal valve

312
313
 Para cont’d---
confirmation of a clinical diagnosis.
The most practical applicable test to identify asymptomatically
shedders of M. paratuberculosis is immunological tests.
The most common immunological tests that are used commonly are
ELISA, CFT, AGID and FAT.
Among them ELISA and AGID tests show most promise.
Gamma interferon test (IFN – γ) also used to diagnose the disease
as tuberculosis
Molecular techniques
The most commonly used molecular technique to detect the
organism in feces or tissue is PCR.
NB. Use of different tests in combination can increase diagnostic
sensitivity.
Differential diagnosis
In cattle paratuberculosis must be differentiated from diseases
which cause chronic diarrhea.
So that paratuberculosis must be differentiate from 314
 Para cont’d---
Intestinal form of tuberculosis
Enteritis
Emeriosis(coccidiosis)
Gastrointestinal helminthiasis
Cu deficiency
Tuberculosis is differentiated from paratuberculosis by allergic test.
Enteritis of different etiology are usually acute and have good
antibiotic and symptomatic treatments response.
The latter two occur principally in younger animals and are
distinguishable on fecal examination for oocysts and helminths eggs.
In Copper deficiency there is alopecia around orbit and is usually an
area problem affecting large numbers of animals and responds well
to the administration of copper.
Treatment
Currently, no antimicrobials are approved for the treatment of
Johne's disease.
M. paratuberculosis is more resistant to chemotherapeutic agents315“in
 Para cont’d---
vitro” than M. tuberculosis.
Because of this treatment is not recommended.
Prevention and control measures
Control of Johne's disease is based on two major principles.
These are
Identification and elimination of infected animals
Prevention of new infections
Identification of infected animals is done using the diagnostic tests
available.
Prevention of new infections depends on
 Improvement of environmental management
Avoiding the introduction of infected animals into the herd
Animals testing positive, particularly those are heavy shedders or
have strong-positive ELISA results, should be sent to slaughter as
soon as economically feasible.
Retesting at least annually should be continued until herd tests
indicate a low (<5%) infection prevalence. 316
 Para cont’d---
Calves, kids, or lambs should be birthed in areas free of manure,
removed from the dam immediately after birth in the case of dairy
cattle.
Bottle-fed colostrum that has been pasteurized or obtained from
dams that test negative must be supplied to them.
Vaccination is also a control strategy to prevent the disease.
In many countries use vaccination of calves less than 1 months of
age.
But it is not as such effective to prevent the disease.
6. Brucellosis (Contagious abortion, Bang's disease)
Brucellosis is a chronic infectious disease of animals and human
beings caused by bacteria of the genus Brucella.
The disease is characterized by abortion, retention of placenta, and
to a lesser extent, orchitis and infection of the accessory sex glands
in males.
Etiology
Brucella is a genus of Gram-negative bacteria named after David317
 Bruce cont’d---
Bruce (1855 - 1931).
In1887, he isolated and identified the causative bacteria,
Brucella(B. melitensis) from the spleen of a soldier who had died
from the infection due to drinking of raw milk of infected goat.
The bacteria are small (0.5 to 0.7 by 0.6 to 1.5 µm), non-motile
non-encapsulated ,non- spore forming , gram negative cocco -
bacilli.
The bacteria are partially acid fast because they are not
decolorized by 0.5% acetic acid in modified Ziehl- Neelsen (MZN)
stain.
They are facultative intracellular bacteria.
Most species of Brucella can grow in media which are enriched by
blood or serum.
Brucella species are aerobic and capnophilic (microaerophilic
requires 5 - 10%Co2) for their growth.
They are non – haemolytic on blood agar.
There are 6 species of Brucella . 318
 Bruce cont’d---
These are
1. B. abortus
2. B. melitensis
3. B. ovis
4. B. suis
5. B. canis
6. B. neotomae
Antigenic structure in Brucella species
In Brucella species there are two types of antigens.
They are
1. B. abortus antigen have a symbol of “A” antigen
2. B. melitensis antigen have a symbol of “ M” antigen
Among the two types of antigens the” M” antigen(melitensis type
antigen) is a highly virulent.
Most species of Brucella shares the above common antigenic
structure with different proportions.
Their antigenic proportion is given below as follows 319
 Bruce cont’d---
B. abortus : A:M=20:1
B. melitensis: A:M=1:20
 B. suis: A:M=2:1
B. melitensis has the highest concentration of “M” type antigen.
Epidemiology
The disease is prevalent in most countries of the world.
It primarily affects cattle, buffalo, bison, pigs, sheep, goats, dogs
elk and occasionally horses.
It is a major cause of abortion in cattle in countries without a
national control program.
The disease in humans, sometimes referred to as undulant fever.
So brucellosis is zoonotic disease.
Undulant fever is a serious public health problem, especially when
it is caused by B. melitensis.

320
 Bruce cont’d---
Diseases, principal hosts of Brucella species and their biotypes
Species Affected animal Diseases Number of
biotypes
B. abortus CATTLE Abortion and orchitis 9
Sheep, goats and pigs Sporadic abortion
Horses Bursitis
B. melitensis GOATS and sheep Abortion 3
Cattle Occasional abortion
B. suis PIGS Abortion, orchitis ,arthritis 4
B. ovis SHEEP Epididmitis in rams and 1
sporadic abortion in ewes
B. canis DOGS Abortion, epididmitis and 1
permanent infertility in males

B. neotomae Desert wood rat Non pathogenic for wood rat 1

and for other mammals.

321
 Bruce cont’d---
Sexually mature animals are susceptible to brucellosis; outbreaks
occur in first-calf heifers, older cows infected but do not abort.
In a herd in which disease is endemic an infected cow typically
aborts only once after exposure.
Subsequent gestations and lactations appear normal.
Because after exposure, cattle become bacteremic for a short period
and develop agglutinins and other antibodies.
But they are latent shedders of the pathogen.
Source of infection
Sick animals with clinical signs of brucellosis( abortion) are source
of infection.
Aborted cow expels the organism together with amniotic fluid,
placenta, fetus and uterine discharges.
The organism can expel together with milk, semen and urine etc.
Natural transmission occurs by ingestion of organisms which are
present in large numbers in aborted fetuses, fetal membranes and
uterine discharges. 322
 Bruce cont’d---
Means of transmission
Brucellosis has the following means of transmissions
Horizontal
Vertical
Horizontal transmission is usually by
Direct contamination
Rare case there is the possibility of introduction of infection by flies,
dogs, rats, ticks, infected boots, fodder and other inanimate objects.
Cattle may ingest contaminated feed and water, or lick
contaminated genitals of other animals.
Transmission may occur by artificial insemination when Brucella-
contaminated semen is deposited in the uterus.
Venereal transmission by infected male to susceptible females
appears to be rare.
But venereal transmission is common in brucellosis of sheep caused
by B. ovis.
Contamination of the udder during milking is also one means of 323
 Bruce cont’d---
transmission.
Brucella may enter the body through mucous membranes,
conjunctivae, wounds, or intact skin in both humans and animals.
Vertical transmission is congenital infection may occur in calves
born from infected dams but its frequency is low.
Pathogenesis
In pathogenicity of Brucella endotoxin plays a very crucial rule.
After the initial invasion of the body, localization occurs initially in
the lymph nodes and spreads to other lymphoid tissues, including the
spleen and the mammary and iliac lymph nodes.
Brucella organism is phagocytized by macrophages and neutrophils
in an effort by the host to eliminate the organism.
However, once inside the phagocyte, Brucella is able to survive and
replicate.
The phagocyte migrates via the lymphatic system to the draining
lymph node, where Brucella infection causes cell lysis.
Because of vascular injury, some of the bacteria enter the 324
 Bruce cont’d---
bloodstream and subsequent bacteremia occurs, which disseminates
the pathogen throughout the body.
However, Brucella organism has a predilection for the pregnant
uterus, udder, testicle and accessory male sex glands, lymph nodes,
joint capsules, and bursae.
If the infected animal is pregnant, the organism will colonize and
replicate to high numbers in the chorionic trophoblasts of the
developing fetus.
The resulting tissue necrosis of the fetal membranes allows
transmission of the bacteria to the fetus.
The net effect of chorionic and fetal colonization is abortion during
second half of pregnancy.
Sexually immature and other non pregnant cattle can become
infected but they do not become sick.
In the adult non pregnant cow localization occurs in the udder and
the uterus, if it becomes gravid, is infected from periodic bacteremic
phases originating in the udder. 325
 Bruce cont’d---
That is why brucellosis is a disease of sexually matured and
pregnant animals.
Because even if the animal is infested by Brucella species when it
was young it never appear clinically(the bacteria exist
interacellularly in macrophages in above mentioned organs) until
the infested animals become sexually matured and pregnant.
This is because of allantoic factors that stimulate the growth of
most Brucella spp.
These factors include
1. The presence or absence of alcohol erythritol.
2. The change in concentration of steroid hormones ( progesterone,
estrogen, FSH and LH).
Erythritol is an alcohol produced by the fetus and capable of
stimulating the growth of B. abortus.
It occurs naturally in greatest concentration in the placental and
fetal fluids and is responsible for localization of the infection in
326
 Bruce cont’d---
these tissues.
Invasion of the gravid uterus results in a severe ulcerative
endometritis of the intercotyledonary spaces which results in
abortion.
Clinical signs
Incubation period is 2 – 4 weeks.
If there is no any pregnant animals( mostly heifers) brucellosis may
have latent form.
However, if there are pregnant animals, typical clinical signs of
brucellosis is mass abortion in the second half of pregnancy.
Cattle 5 – 8 months of pregnancy
Sheep and goat 3- 5 months
Pigs may abort in both first and second halves.
Dog 40 – 50 days.
The “storm” might last for a year or more, at the end of which time
most of the susceptible cows are infected and have aborted.
Retained placenta and metritis could be expected to be common 327at
 Bruce cont’d---
this time.
Infection may continue to affect fallopian tube and ovaries.
All this results in infertility of animals.
Brucellosis in animals can be characterized by bursitis, arthritis,
tendovaginitis.
In bulls and rams there can be orchitis and epididymitis in one or
both scrotal sacs.
Orchitis and epididymitis with acute inflammation can be
characterized by painful swelling( twice the normal size).
The seminal vesicles may be affected and their enlargement can be
detected on rectal palpation.
Affected bulls are usually sterile when the orchitis is acute but may
regain normal fertility if one testicle is undamaged.
Such bulls are potential spreaders of the disease if they are used for
artificial insemination.
In pigs in addition of the above clinical signs there is abscess in
subcutaneous tissue and in many parenchymatous organs. 328
 Bruce cont’d---
In horses brucellosis is characterized by bursitis.
Necropsy findings
The fetal membrane always edematic and is covered by fibrin and
pus.
Necrotizing placentitis and placenta is usually edematous.
There may be leathery plaques on the external surface of the
chorion and there is necrosis of the cotyledons.
The key microscopic feature of this inflamed chorioallantois is the
presence of intracytoplasmic coccobacilli within chorionic
trophoblasts.
The use of modified Ziehl-Neelsen stains on impression smears
from fresh placentas can provide a rapid presumptive diagnosis of
brucellosis.
There may be signs of metritis with cataract and pus; ovaritis with
development of cysts in the ovaries.
You may also observe mastitis, arthritis and bursitis.
329
Gross pathology of cows and fetuses experimentally infected with
Brucella abortus.

330
Placentitis on cotyledon of ruminants due to brucellosis

331
Carpal bursitis and arthritis in calves due to B. abortus

332
 Bruce cont’d---
There may be abscess in liver, spleen and kidneys.
In male animals you may observe testicular abscesses.
Disseminated inflammatory reactions in aborted fetal tissues are
the characteristic changes like
Edema of subcutaneous tissue and umbilicus
 Sero –hemorrhagic fluid with fibrin in the body cavities and
subcutis.
Haemorrhagy on serose and mucous membranes.
 Cataract inflammation of mucous membrane of GIT, lung with
necrotic foci in liver.
Diagnosis
Diagnosis of brucellosis is done by complex method by considering
the
Epidemiology of the disease
Clinical findings
Allergic test
333
 Bruce cont’d---
Pathological changes
Laboratory( bacteriology and serology)
In epidemiological diagnosis consider
The serological and/ or allergic test result of the herd for brucellosis
in last year.
The presence of prophylactic quarantine for a new animals
The presence of mass abortion in the second half of pregnancy.
The average calving rate and calving interval of the farm etc.
In clinical diagnosis consider some of like
The presence of mass of bursitis, arthritis and orchitis, endometritis,
retention of placenta etc.
In necropsy findings consider some of like
 The presence of necrotizing placentitis with mass of abortion in the
second half of pregnancy.
The presence of leathery plaques on the external surface of the
chorion and necrosis of the cotyledons.
The key microscopic feature of this inflamed chorioallantois is the 334
Brucella in MZN staining clubs of intercellular bacteria in
macrophages

335
 Bruce cont’d---
presence of intracytoplasmic coccobacilli within chorionic
trophoblasts that can be confirmed by modified Ziehl-Neelsen stains
on impression smears.
There are many additional diagnostic methods of brucellosis.
Among them the most common and practical applicable are
Allergic
Serological
Bacteriological
Molecular method
Allergic method
This method is less commonly used as compared the rest two
methods.
The principle of allergic test is based on delayed hypersensitivity test
to screen out infected animals from the herd.
The allergen that is commonly used to diagnose brucellosis is called
brucellin.
Brucellin can be injected 336
 Bruce cont’d---
 S/C around the skin of lower eye lid( sheep, goat and cattle)
 Intradermal in external surface of ear( pigs)
Dose and sites of injection of brucellin in different animals

Species of animals Injection sites Dose

Cattle S/C in lower eye lid/ 1 ml S/C/ 0.3 ml intradermal
intradermal caudal folds
Sheep and goat S/C in lower eye lid/ 0.5 ml S/C/ 0.2 ml intradermal
intradermal caudal folds

Pigs Intradermal into the skin of 0.2 ml

external ear

Test result is recorded after 36 – 48 hours.

The presence of inflammation at the site of injection is considered
as positive.
This method is not commonly used in our country
NB. Do not inject brucellin to the lower eye lid if there is trauma
and abscess. 337
 Bruce cont’d---
Serological methods
Immunological tests for detecting antibodies to Brucella abortus
Since brucellosis is a chronic infection disease, it is important to
diagnose the disease in live animals to control the disease in your
herd.
The presumptive serological diagnosis is usually made based on the
presence of antibodies in serum, milk, whey, vaginal mucus, or
seminal plasma.
A wide range of immunological tests has been developed to
diagnose brucellosis in live animals.
The principle of the immunological tests is on the detection of
Antibodies(IgM and Ig G) that are formed after natural infection.
However the Ig M titer is not long lasting that means
immunological tests of brucellosis is based on the detection of Ig G.
Immunological tests used to diagnose brucellosis can be screening
and confirmatory.
Among immunological screening tests the most common are 338
 Bruce cont’d---
1. Brucella Serum Agglutination Test(SAT)
2. Rapid Plate agglutination test (Rose – bengal plate test)
3. Brucella milkring test
1. Brucella Serum Agglutination Test(SAT)
It is non – specific serum agglutination test that detects non-
specific antibodies as well as those that are specific for Brucella
abortus infection and vaccination.
Since it has low specificity it is not widely used for diagnosis of
brucellosis.
2. Rapid Plate Agglutination Test (Rose – bengal plate test)
Rapid plate agglutination test is also screening test to diagnose
brucellosis in animals.
The principle of the test is that the latex materials or red blood
cells are conjugated with Brucella antigen.
If the testing serum contains antibody against Brucella antigen,
they agglutinate and clumps are visible on slide.
339
 Bruce cont’d---
The most common rapid plate agglutination test which is used to
diagnose brucellosis is Rose - Bengal plate agglutination test.
Rose - Bengal plate agglutination test
It is serological screening test to detect the presence of the disease
in our herd by detecting specific antibody (Ig G) against B.
abortus.
It is screening test because even if it has high sensitivity, the
specificity of the test is too low.
Since Rose bengal plate agglutination test has high sensitivity but
low specificity, there is false positive test result.
Because there are many gram negative bacteria which share
common somatic antigen with B. abortus.
Among them the most common species of bacteria which share
common somatic antigen with B. abortus is Yersinia
enterocolitica(Somatic antigen O – polysaccharide(0:9)).
Other gram negative bacteria which share common somatic
antigen with B. abortus and can cause false positive result in Rose 340
341
342
343
 Bruce cont’d---
Bengal plate agglutination test are
 1.Genus Francisella
 2.Genus Campylobacter
 3.Genus Salmonella
 4 . E. coli(O116:H21 and O157:H7)
 5. Vibrio cholerae
 4.Genus Pasteurella
That is why Rose bengal test is screening test and those that are
positive in Rose bengal test should be retested by one of
confirmatory tests.
3. Brucella milk ring test
The milk ring test is a satisfactory inexpensive test for the
surveillance of dairy herds for brucellosis.
Brucella milk ring test is a screening test for brucellosis infection at
the herd level by taking a small sample of pooled fresh milk or
cream, from no more than 25 cows.
The test is based on the principle of agglutination test that can be344
 Bruce cont’d---
carried out with body fluids other than serum that is with milk.
In the milk ring test stained Brucella abortus organism with
methylene blue dye or Rose – bengal dye is used as suspension of
antigen.
The principle of the test is that if the organism is chronically
affected by brucellosis the antibody which is secreted together with
milk will react with stained Brucella antigen and blue ring will be
formed at the top where the cream is located.
Confirmatory serological tests
Among confirmatory immunological tests that are used to diagnose
brucellosis, the most common are
1. Complement Fixation Test(CFT)
2.Enzyme Linked Immuno Sorbent Assay(ELISA)
3. Agar gel – Immuno Diffusion (AGID)
NB. The serological diagnosis is considered to be unreliable when
applied during the period of 2 - 3 weeks before and after abortion
or calving. 345
346
 Bruce cont’d---
NB. False-negative test result may occur and can be detected by
retesting animals at intervals over a period of at least 3 months.
Screening and confirmatory tests for different species of Brucella
Species of Brucella Screening test Confirmatory test
B. abortus Rose Bengal, (milk CFT or ELISA
ring test at herd level)
B. melitensis Rose Bengal or Serum CFT or Agar Gel Immuno
agglutination test Diffusion(AGID) or ELISA
B. suis Rose Bengal test CFT or ELISA
B. ovis Rose Bengal test CFT or ELISA
B. canis Rapid slide CFT or ELISA or
agglutination test Agar gel immuno diffusion
test(AGID)

347
 Bruce cont’d---
Bacteriological method of diagnosis
It is confirmatory diagnostic method based on the isolation and
identification of the positive agents.
In abortion cases a full range of specimens should be collected and
submitted for laboratory diagnosis.
The specimens that must be collected are
 A whole fetus if it is feasible.
 Alternatively, fetal stomach contents, any fetal lesions, cotyledons,
uterine discharges, urine( for leptospirosis), colostrum and paired
serum samples.
 Semen and tissue from epididymides or testes from males must be
submitted to laboratory.
Differential diagnosis
Brucellosis must be differentiated from many infectious and
parasitic disease that are accompanied with abortion like
 Campylobacteriosis
 Trichomoniasis 348
 Bruce cont’d---
 Epididymitis infectious and non – infectious etiology
 Listeriosis
 Leptospirosis
 Chlamydiosis with abortion
 Abortion non – infectious etiology
Treatment
Treatment is unsuccessful because of the intracellular sequestration
of the organisms in lymph nodes, mammary gland and reproductive
organs.
So that there is no any antimicrobial agent that can penetrate the
cell membrane of cells.
Prevention and control measures
In farms free of brucellosis strict veterinary sanitary measures must
be undertaken.
That includes
Prevention of the farm no to be infected by other animals in pasture
and water source. 349
 Bruce cont’d---
Replacement animals must be from brucellosis free farms
Quarantine of new replacement animals for at least 30 days and mix
with other animals after consequentive serological tests( all tests
must be negative).
Use semen from bulls that are free from brucellosis( natural mating
or artificial insemination)
In every six months the farm must be tested for brucellosis.
If there are mass abortion, appropriate specimens must be submitted
to referral laboratory.
Then if there is mass abortion due to brucellosis, the following
veterinary sanitary practice must be done to eradicate the infection.
These are
1.Quarantine
2. Depopulation
3. Vaccination
1. Quarantine
This is a period of time during which cattle movement is restricted
350
 Bruce cont’d---
and the cattle are tested.
This will prevent interherd transmission by infected cattle,
especially those that are test-negative and incubating the disease.
The quarantine period should be sufficiently long that all cattle
have had sufficient time to develop brucellosis and insure that the
remaining cattle will not be a source for interherd transmission.
The quarantine time for brucellosis usually range from 120 days
to 1 year or
Until all breeding animals have completed a gestation without test
evidence of infection.
2. Depopulation
Depopulation is slaughter of all cattle in a herd when all animals
have been exposed and are capable of becoming infected and
acting as a source of new infection.
This means test and reduction of carriers of infection.
Eradication of these infected cattle helps to reduce the economic
losses and to protect the public from the disease. 351
 Bruce cont’d---
Test and reduction of reservoir of infection.
Herds must be tested at regular intervals until 2 or 3 successive tests
are negative.
3. Vaccination
Vaccination is not commonly used in most countries of developing
world.
However in some countries use B. abortus strain 19 or RB51
vaccine to prevent bovine brucellosis.
Vaccination of calves with B. abortus strain 19 or RB51 increases
resistance to brucella infection.
The optimum age for vaccination is between 4 and 8 months
There is no significant difference between the immunity conferred
at 4 and at 8 months of age.
Also there is whole-herd adult cattle vaccination using strain 19 or
RB51 has been practiced in certain high brucellosis -incidence
areas.
352
7. Pasteurellosis
It is bacterial disease of several species of domestic animals
including poultry characterized by
 Haemorrhagic septicemia in acute form
Pneumonia in subacute and chronic form
Etiology
Pasteurellosis in domestic animals is caused by two species
Pasteurella multocida
Mannheimia haemolytica (formerly Pasteurella haemolytica)
They are small( 0.2µm by up to 2µm),Gram – negative rods or
coccobacilli.
They are non - motile, non- spore forming facultative anaerobe.
They are oxidase and catalase positive.
They can grow on/ in non- enriched media like nutrient agar and
broth , Trypticase Soya Agar (TSA), Trypticase Soya Broth(TSB)
etc.
But their growth is enhanced on/ in media contains blood or serum.
Giemsa staining on smears prepared from affected tissues and 353
 Pastur. cont’d---
blood during septicemia, the Pasteurella (Mannheimia) spp have
bipolar staining character.
On blood agar most of the Pasteurella spp. colonies are haemolytic
except the colonies of P. multocida and P. pneumotropica.
Most of the species of Pasteurella including Mannheimia
haemolytica grow on MacConkey agar and are non- lactose
fermenter.
But P. multocida never grow on MacConkey agar.
All species of Pasteurella including Mannheimia haemolytica are
extracellular parasite and commensals on mucous membrane of the
upper respiratory tracts.
Infections caused by these organisms are commonly associated with
stress factors( concurrent infections and parasitic diseases,
transport, feed and water shortage and environmental and
microclimatic conditions etc.).
Even though there are several species of Pasteurella that can exist as
commensals in upper respiratory tracts of different animals, the most
354
 Pastur. cont’d---
common species of that have veterinary importance are
Pasteurella multocida
Mannheimia haemolytica
By their capsular antigen differences P. multocida has the following
serogroups
Serogroup A
Serogroup B
 Serogroup D
 Serogroup E
Serogroup F
There is a relationship between the capsular subgroup and disease
predilection.
They are also different in somatic antigen structure(LPS).
Somatic antigens of Pasteurella multocida are represented by
numbers.
So the serotype is identified by serogroups followed by its somatic,
type. 355
The principal hosts and diseases caused by Pasteurella multocida
and Mannheimia haemolytica
Spp. Of Pasteurella Serogroup Principal Disease
and Mannheimia host (s)
Shipping fever,
Cattle Pneumonic pasteurellosis
A Occasionally but sever mastitis
Sheep Plueropneumonia and mastitis.

Poultry Fowl cholera

B Cattle Haemorrhagic septicemia( barbone) in
south east Asia.
P. multocida
D Pigs Pneumonic form pasteurellosis
Atrophic rhinitis( with or with out
Bordetella bronchiseptica)
E Cattle and Haemorrhagic septicemia( barbone) in
buffalo Africa only.
F Turkeys Isolated in turkeys mainly but its role in
mainly disease is not known.
356
 Table Cont’d---
Spp. Of Pasteurella Serogroup Principal host Disease
and Mannheimia (s)
Cattle Shipping fever
Pneumonia pasteurellosis
Mannheimia A Sheep and Pneumonia pasteurellosis
haemolytic goat Septicemia in lambs under 3
months of age.
Gangrenous mastitis
T Sheep Septicemia in lambs 5 – 12
months old.

357
 Pastur. cont’d---
For example the cause of bovine hemorrhagic septicemia is B:3 in
in south east Asia and E:6 in Africa.
The two serogroups of M. haemolytica have different
pathogenicity, antigenic nature and biochemical activity.
For example M. haemolytica serogroup A ferments arabinose and
serogroup T ferments trehalose.
The serotype of M. haemolytica is determined by its somatic
antigen (lipopolysaccharides).
So the serotype is identified by numbers.
For example M. haemolytica that cause pneumonic pasteurellosis
of cattle in Europe is Mannheimia haemolytica biotype A.
Diseases associated with Pasteurella and Mannheimia species
The most common diseases of domestic large animals that are
caused by Pasteurella and Mannheimia species are
 1. Pneumonic pasteurellosis
 2. Septicemic pasteurellosis
 3. Haemorrhagic septicemia in cattle 358
 Pastur. cont’d---
 4. Bovine pneumonic pasteurellosis(shipping fever)
 5.Atrophic rhinitis in pigs
1. Pneumonic pasteurellosis in sheep
Pneumonic pasteurellosis in sheep is usually caused by M.
haemolytica.
However, P. multocida tends to produce sporadic cases of the
disease.
You know that M. haemolytica is a commensals of the upper
respiratory tract in a proportion of healthy sheep.
Factors which predispose to clinical disease are poorly understood
and may include
Adverse climatic conditions and/or
Concurrent infections with respiratory viruses and lung parasites
such as parainfluenzavirus – 3 and/ or lung worms etc.
Flock outbreaks usually start with sudden deaths of some sheep and
acute respiratory distress in others.
359
 Pastur. cont’d---
2. Septicemic pasteurellosis in sheep
Septicaemic pasteurellosis in lambs less than 3 months of age is
caused by M. haemolytica(M. haemolytica type A).
In older animals between 5 and 12 months of age , septicemic
pasteurellosis is usually associated with P .trehalosi(M. haemolytica
type T).
M. trehalosi is found in the tonsilar tissues of carrier sheep.
As with most other pasteuurella infections, clinical disease may be
precipitated by the range of predisposing factors including
transportation.
Affected sheep which tend to be in good body condition may die
suddenly and the mortality rate may approach 5%.
3. Haemorrhagic septicemia in cattle
Haemorrrhagic septicemia or barbone is an acute, potentially fatal
septicemia mainly affecting buffalo and cattle.
The predisposing factors that are important for its development are
 Overwork 360
 Pastur. cont’d---
 Poor body condition
 Radical seasonal fluctuation
P. multocida serotype: 2(B:6) causes the disease in Asia, Middle
East and some southern European countries.
While serotype E:2(E:6) is the cause of hemorrhagic septicemia in
Africa.
These serotypes are the only serotypes of P. multocida with
hyaluronidase.
Buffaloes tend to be more susceptible to the disease than cattle.
All age groups can be affected but in endemic areas the disease is
most common in animals between 6 and 24 months of age.
Older animals may have immunity from previous exposure.
Many older animals are latent carriers with the organism located in
the tonsillar crypt.
Periodically these animals shed P. multocida in nasal secretions and
in aerosols.
Explosive outbreaks of pneumonic pasteurellosis can occur if an361
 Pastur. cont’d---
active carrier is introduced into a stressed and susceptible
population.
4. Bovine pneumonic pasteurellosis(shipping fever)
Shipping fever characterized by severe broncho- pneumonia and
pleurisy occurs most commonly in young cattle within weeks of
being subjected to severe stress such as: -
 Transportation
 Assembly in feedlots
 Close confinement
 The presence of concurrent viral infections like parainfluenzavirus
3, bovine herpesvirus1 and bovine respiratory syncytial virus
Bovine pneumonic pasteurellosis( shipping fever) is commonly
associated with M. haemolytica.
However, P. multocida has also been isolated from lungs of affected
cattle.
The principal serotype of M. haemolytica associated with
pneumonic form of pasteurellosis in cattle are 362
 Pastur. cont’d---
 Sero type A1
 Serotype A6
5.Atrophic rhinitis in pigs
Toxigenic strains of P. multocida type D or A cause a severe
progressive form of atrophic rhinitis.
These toxigenic strain of P. multocida isolates are designated AR+(
atrophic rhinitis – positive) strains.
The predisposing factors to infection are
 Infection with Bordetella bronchiseptica
 Over – stocking
 Poor ventilation
Epidemiology
Pasteurellosis affect almost all species of animals including poultry.
Animals of all ages are susceptible but the most susceptible age
group is 6 months to 2 years of age( in cattle).
There is no difference in susceptibility between breeds.
Dogs and horses are comparatively resistant species. 363
 Pastur. cont’d---
The disease occurs as epizootics( epidemics) in poultry but rarely in
other species of animals.
In other species of animals pasteurellosis occurs as epizootics(
haemorrhagic septicaemia) if the disease is exotic.
But in area where pastreullosis is endemic the disease occurs
mostly sporadically in pneumonic form.
The incidence of disease is reduced significantly in areas where the
vaccine is used.
Both morbidity and case-fatality rates vary between 50% and 100%
respectively.
Morbidity will depend on the immune status of the herd, either
acquired naturally or induced by vaccination.
Outbreaks of the disease are often associated with wet humid
weather during the rainy season and other stress factors.
Source of infection
The source of infection can be
Clinically sick animals 364
 Pastur cont’d---
Clinically normal carrier animals
These animals expel the pathogen with
Nasal discharge
Saliva and exhaled gases
Faces
Mechanism of transmission
The pathogen spreads by
 Contaminated feed
Aerogenic
Epidemiological character of pasteurellosis is its enzootic character.
Morbidity and mortality of pasteurellosis depends on
Serogroup and serotype of Pasteurella species
Virulence of bacteria
Immunological situation of the herd
Managemental conditions of animals( feeding and housing)
 The presence or absence of stress factors( concurrent infection or or
invasion, transportation etc).
365
 Pastur. cont’d---
The time when control measures are done
Pathogenesis
Infection can be endogenous or exogenous.
The route of infection is usually via respiratory tract although the
bacteria can infect susceptible animals by ingestion and through
trauma.
A fulminating septicemia occurs, which is associated with the
capsular material of the organism.
Virulence of Pasteurella is enhanced by animal to animal
transmission.
The bacteria reproduce at the place of entery and enters into the
lymph and blood systems.
Endotoxins are particular important in pathogenicity of virulent
strain of Pasteurella species.
It inactivate phagocytic cells, damages capillaries and causes edema
in subcutaneous tissues of skin and in intramuscular fats.
366
 Pastur. cont’d---
All these lead to the development of hemorrhagic septicaemic form
of pasteurellosis.
Septicemic form of pasteurellosis is formed when the pathogen is
virulent.
However, when less virulent species Pasteurella infect animals with
good resistance septicemic form of pasteurellosis is not develop.
Rather the bacteria will localized in some organs specially in the
lung tissue and cause fulminating fibrinous lobar pneumonia or
fibrinopurulent bronchopneumonia(pneumonic form of
pasteurellosis).
Some strains of Pasteurella multocida( strain type “D “) produce
thermo liable dermonecrotising toxin which have an important role
in atrophic rhinitis of pigs.
We know that the colonization of P. multocida strain type “D’ is
facilitated by Bordetella bronchiseptica.
The virulence factor of Mannheimia haemolytica is associated with
its ability to produce cytotoxin( leukotoxin). 367
 Pastur. cont’d---
Leukotoxin has a role to over come the lung primary defense
mechanism by its action on the alveolar macrophages and other
leucocytes.
Clinical signs
1. Septicaemic form of pasteurellosis has the incubation period of 2
– 4 days.
Death, without prior signs of illness may occur with in 24 hours of
infection.
It is clinically characterized by
 Sudden onset of fever (41- 42°C)
Severe respiratory distressed may occur if the respiration is
obstructed by respiratory discharges
Localization may occur in subcutaneous tissue, resulting in the
development of warm, painful swellings about the throat, dewlap,
brisket or perineum.

368
 Pastur. cont’d---
In the later stages of an outbreak, some affected animals develop
signs of pulmonary or alimentary involvement.
Mortality rates are usually above 50%.
2. In pneumonic form of pasteurellosis; in the first few days of an
outbreak a number of sheep will show clinical signs of pneumonia.
The clinical features are
Sudden onset of fever
Depression
Anorexia
Tachypnoae
Serous nasal discharge
In mixed infections, there is usually a marked cough and ocular
discharge.
3. In Atrophic rhinitis of pigs
Early signs usually encountered in pigs between 3 and 8 weeks age.
The early clinical signs includes
Excessive lacrimation 369
 Pastur. cont’d---
Sneezing and occasionally epitaxis
The snout gradually becomes shortened and wrinkled
As the disease progress, a distinct lateral deviation of the snout
may develop.
Atrophic rhinitis is rarely fatal.
Affected pigs are usually underweight and damage to the turbinate
bones may predispose to secondary bacterial infections of the lower
respiratory tract.
Necropsy findings
In septicemic form of pasteurellosis there is generalized petechial
hemorrhages particularly under the serosae
 Edema of the lungs and lymph nodes.
Subcutaneous infiltrations of gelatinous fluid may be present
 In a few animals there are lesions of early pneumonia and a
hemorrhagic gastroenteritis.
In pneumonic form of pasteurellosis the post mortem findings is
370
located on cranial ventral lobes of lungs that become ventral red,
 Pastur. cont’d---
swollen and fibrinous pleural with epicardial effusions.
There may be thickening of the interlobular septa.
Lymph nodes in the thoracic region are enlarged and hemorrhagic
In atrophic rhinitis of pigs, there is
Facial deformities
Atrophy of turbinate cause the snout to be shortened and found
between the first and second premolar teeth.
Diagnosis
Diagnosis of pasteurellosis is done based on
History of exposure to stress
Clinical signs have no typical pathognomonic signs and have little
value in diagnose of pasteurellosis.
However, if you made blood smear in septicemic form of
pasteurellosis; stained in Giemsa and found bipolar organism, You
could suspect septicemic form of pasteurellosis
Necropsy findings
Laboratory diagnosis 371
 Pastur. cont’d---
Gross pathological findings like the presence of red swollen lung
tissue on cranial ventral lobes of lungs with fibrinous pleural with
epicardial effusions may help to suspect pneumonic form of
pasteurellosis.
Laboratory is confirmatory diagnose.
Suitable specimens for laboratory examination from live animals
include
Tracheobronchial aspirates
Nasal swabs or mastitic milk
Tissues and blood smears from septicemic cases, stained by Giemsa
may reveal large numbers of bipolar – staining organisms.
Specimens should be cultured on blood agar and MacConkey agar.
Blood agar, supplemented with neomycin, bacitracin and actidione
can be used for the isolation of P. multocida heavily contaminated
specimens.
In atrophic rhinitis severely affected pigs characteristic facial
deformities are diagnostic. 372
 Pastur. cont’d---
Isolation and identification of P. multocida should be followed by
tests to confirm that the isolate is a toxigenic strain by ELISA and
detection of toxin gene by PCR.
Differential diagnosis
Pasteurellosis in cattle and sheep must be differentiated from
Anthrax
Piroplasmosis
Black leg
Staphylococcal and streptococcal infections
Salmonellosis
Colibacteriosis
Respiratory virus infections like Parainfluenza – 3 and infectious
rhinotrachitis
Pasteurellosis in pigs must be differentiated from
Swine pests
Erysipelas
373
Salmonellosis
 Pastur. cont’d---
Treatment
Affected animals must be isolated and treated early in the course of
the disease.
Treatment with oxytetracycline, potentiated sulphonamides and
ampicillin is usually effective.
Prevention and control measures
Stress factors must be kept to a minimum.
Procedures such as castration, dehorning, branding and
antihelmintics therapy should be carried out several weeks before
young cattle and sheep are transported.
Have good housing with good microclimatic condition and avoid
overstocking.
Latent carriers can be detected using immunohisto - chemical
techniques on samples of tonsillar tissues.
Vaccination regimes for respiratory pathogens should be completed
at least 3 weeks before transportation.
374
Vaccines available for control of the disease, include bacterins and
 Pastur. cont’d---
a modified live vaccine.
Recently, a vaccine containing serotype – specific antigens of both
M. haemolytica A1 and M. haemolytica A6 has been developed in
Europe.
Colostral immunity of calves from cows vaccinated against
hemorrhagic septicemia peaks at 8 -16 weeks of age and then
declines.
NB. In Ethiopia, there is formalin inactivated aluminum hydroxide
adjuvant vaccine for both cattle and sheep.
Dose of vaccine
 Cattle 2ml S/C
Sheep 1 ml S/C
Immunity lasts 6 - 8 months.
Bacterial diseases caused by Enterobactericeae
Here we will see
8. Colibacillosis
9. Salmonellosis 375
 Coli cont’d---
8. Colibacillosis
It has another names like “Escherichiosis “or “coli – infection” or
colibacteriosis.
It is acute bacterial diseases of calves, lambs and piglets
characterized by profuse diarrhoea, sign of intoxication and
dehydration in calves and lambs; coli – enterotoxemia with sign of
edema in piglets just after weaning.
Etiology
The disease is caused by pathogenic strains of E. coli.
E. coli is gram negative rod shaped, facultative anaerobe, non –
spore forming and most pathogenic strains are flagellated and
capsulated.
They grow in/on conventional media like blood agar.
They grow on selective media like Xylose Lysine
Decarboxylase(XLD medium) and brilliant green agar.
Most strains of E. coli are normally considered as pathogenic for
new born animals.
In adult animals they cause opportunistic infections in site of the 376
 Coli cont’d---
body such as mammary gland (mastitis) and uterus (metritis).
The diseases of new born animals are caused by pathogenic
serotypes of Escherichia coli.
Surface antigens of E. coli
E. coli has the following antigens: -
 1.Capsular (K) antigens which is polysaccharides.
2.The cell wall or somatic (O) antigens are determined by the sugar
side – chains on the lipopolysaccharide molecules of the outer
membrane.
3. Flagellar ( H) and fimbrial(F) antigens which are proteins.
Some of the most well known Fimbrial(F) antigens are K88(F4)
and K99(F5).
Fimbrial antigens allow pathogenic E. coli strains to adhere to
intestinal cells of small intestine.
The O, K and H antigens are used to serotype strains of E.coli.
377
Each serotype is designated by the number of antigens that it bears.
 Coli cont’d---
For example O157:K85:H19.
Epidemiology
Colibacillosis occurs most commonly in newborn farm animals
and is a significant cause of economic loss in raising livestock
specially in intensive farms.
Risk factors
The common risk factors are
1. Host and pathogen risk factors
2. Environmental and management risk factors
Rotavirus, Cryptosporidium spp, corona virus and enterotoxigenic
E. coli collectively are responsible for 75 - 95% of infections in
neonatal calves worldwide.
So that age of animals and associated serotypes of E. coli are the
crucial risk factors.
Enterotoxigenic colibacteriosis is common in calves and lambs
under 3 days of age.
The prevalence can be as high as 50 - 60% in diarrheic calves 378
 Coli cont’d---
under 3 days of age and only 5 - 10% in diarrheic calves at 8 days of
age.
Diarrhea caused by enterotoxigenic E. coli occurs in calves mainly
during the first 10 days of life and very rarely in older calves, and
never in adults.
In piglets the prevalence of enterotoxigenic E. coli in diarrheic
piglets varies geographically and with herds.
Other risk factors is the microclimatic condition of housing and
management
Predisposing factors are of paramount importance to cause clinical
illness of animals by E. coli.
Among them the most important are:
1.Insufficient( qualitatively or quantitatively) obtaining of new born
animals neonates) passive immunity( antibodies) with colostrum.
It is known that the transfer of maternal immunoglobulins to calves
depends on three successive processes those are
Formation of colostrum with a high concentration of
379
 Coli cont’d---
immunoglobulin by the dam
Ingestion of an adequate volume of colostrum by the calf
 Efficient absorption of colostral immunoglobulins by the calf
Colostral immunoglobulins are absorbed for upto 24 hours afterbirth
in calves and upto 48 hours in piglets.
However, in calves maximum efficiency of absorption occurs during
the first 6 -12 hours after birth and decreases rapidly from 12 - 24
hours after birth.
2. Intensive husbandry practice led to the rapid transmission of
pathogenic strains of E. coli strain.
3. Poor hygienic condition creates favorable condition for
pathogenic strains of E. coli in the young animal’s environment
because a large dose of pathogenic E. coli may over come clostral
immunity in young animals.
Stress factors such as cold ambient temperatures and frequent
mixing of animals may predispose new born animals to coli –
infection. 380
 Coli cont’d---
New born animals under one week of age are always susceptible to
E. coli infection; this is because:-
The normal flora of the intestine is not fully established.
They have a “naïve” immune system(Th 0 cells).
Receptors for fimbrie of E. coli is present only in the first week life
Digestive tract of young pigs equipped only for easily digested
foods so that accumulation of undigested and unabsorbed food
encourage replication of E. coli in GIT of piglets.
Pathogenesis
E. coli strains that cause enteritis in new born animals have been
classified as: -
1. Enterotoxigenic E. coli (ETEC)
2. Enteropathogenic E. coli (EPEC)
3. Enteroinvasive E. coli ( EIEC)
4. Attaching and Effacing E. coli ( AEEC)
5. Necrotoxigenic strains of E. coli
381
 Coli cont’d---
1. Enterotoxigenic E. coli (ETEC)
In this group belongs E. coli with fimbrie adhesin.
They produce enterotoxin after attachment which can be
Heat labile(LT) which causes hypersecretion in mucousa of
intestine.
Heat – stable(ST) which reduces absorption but enterocytes remain
in structure.
They cause the majority of neonatal colibacillosis( diarrhea in
neonatal piglets, calves and lambs) and post weaning diarrhea in
pigs.
The most common strains of E. coli that are ETEC are E. coli with
K88 or K99 antigen.
2. Enteropathogenic E. coli (EPEC)
Enteropathogenic Escherichia coli (EPEC) does not produce
enterotoxin but induces severe watery diarrhoea.
This is linked with weak inflammatory response.
Because EPEC flagellin triggers the secretion of the 382
 Coli cont’d---
inflammatory mediators called cytokine like interleukin (IL – 8) etc.
EPEC strains are isolated from lambs with diarrhea.
3. Enteroinvasive E. coli ( EIEC)
Strains which adheres to enterocytes of the distal part of small
intestine.
They invade enterocytes and the deeper layers of intestinal mucosa
by its virulence factor adhesin and invasin.
After this they reach the lymphatic system of intestine where they
multiply.
On the way some the bacteria may die and release endotoxin.
The important virulence factor for survival for this group of E. coli
are: -
Capsule
Adhesin
Siderophores
Alpha haemolysin
This group of invasive E. coli are responsible for colisepticemic 383
 Coli cont’d---
colibacillosis in calves and piglets.
4. Attaching and Effacing E. coli ( AEEC) or Verotoxigenic E.
coli(VTEC)
It has also another names like enterohemorrhagic E. coli (EHEC)
also referred to as Verotoxigenic E. coli(VTEC) or Verocytotoxigenic
E. coli (VTEC) or shiga-toxin producing E. coli(STEC).
These strains of E. coli which binds to enterocytes of small
intestine and damage them by producing shiga toxins called
verotoxins(VT1,VT2 and VT2e).
The shiga - like toxins( vero toxins) destroy the microvillus by
unknown means.
This strain of E. coli cause edema disease in piglets, hemorrhagic
enterocolitis in calves and post weaning diarrhoea in piglets.
E. coli 0157:H7, one of the important enterohemorrhagic E. coli
strains.
384
 Coli cont’d---
5. Necrotoxigenic strains of E. coli
This strain of E. coli binds to enterocytes and produce cytotoxic
necrotizing factor like CNF1 and CNF2.
These toxins damage to enterocytes and blood vessels and cause
hemorrhagic inflammation and cause:-
 Haemorrhagic colitis in calves
 Enteritis in calves and piglets
Edema disease in piglets is caused by E. coli withO139 and O141
strains that produce a verotoxin(VT2e) just after weaning.
Verotoxin is absorbed into the blood stream and damages
endothelial cells with consequent perivascular edema.
NB. All pathogenic groups of E. coli are haemolytic on blood agar.
Clinical findings
1. Calves with septicemic colibacillosis
It is most common during the first 3 days of life.
Illness is acute, the course varying from 24 - 96 hrs.
There are no typical diagnostic clinical signs but affected animals
385
 Coli cont’d---
are
Depressed and weakened
Anorexia is complete with marked tachycardia
Initially temperature is high and drops with time
Oral mucous membrane is dry and cool
If the calf survives the septicemic state, clinical evidence of post
septicemic localization may appear in about 1 week.
This include
Arthritis
Meningitis
 Pneumonia (less commonly)
2. Calves with enterotoxigenic colibacillosis
It is most common form of colibacillosis in calves from 3 to 5 days of
age.
In this form of colibacillosis the outstanding signs include
Severe weakness
Coma
Subnormal temperature 386
 Coli cont’d---
Pale mucosae
Wetness around mouth
Prognosis for these calves is poor they commonly die within 2 - 6
hrs after the onset of signs.
3. Calves with enteric colibacillosis
This is the most common form of colibacillosis in newborn calves
primarily3 - 5 days of age.
It may occur in calves as early as 1 day of age and only rarely up to
3 weeks.
The clinical severity will vary depending upon the number and kind
of organisms causing the disease.
The presence of a single enterotoxigenic strain of E. coli may cause
a state of collapse usually designated as enteric toxemia.
4. In lambs and goat kids
Although some cases manifest enteric signs, and chronic cases may
occur, colibacillosis in lambs is commonly septicemic and peracute.
Two age groups appear to be susceptible lambs 1 - 2 days of age387and
 Coli cont’d---
lambs 3 - 8 weeks old.
Peracute cases are found dead without premonitory signs.
Acute cases show collapse and occasionally signs of acute
meningitis manifested by a stiff gait in the early stages, followed by
recumbency with hyperesthesia and titanic convulsions.
5. Piglets coliform septicemia
This is uncommon but occurs in piglets within 24 - 48 hours of birth.
Some are found dead without any premonitory signs.
Usually more than one, and sometimes the entire litter, are affected.
Severely affected piglets seen clinically are weak, almost comatose,
appear cyanotic, and feel cold and clammy and have a subnormal
temperature.
Usually there is no diarrhea.
The prognosis for these is poor and most will die in spite of therapy.
Necropsy findings
In coliform septicemia there may be no gross lesions and the
diagnosis may 388
 Coli cont’d---
depend upon the isolation of the organism from the filtering
organs(spleen and lymph nodes ).
In less severe cases there may be subserosal and submucosa
hemorrhages.
A degree of enteritis and gastritis may be present.
Occasionally, fibrinous exudates are found in the joints and serous
cavities, and there may be omhalophlebitis, pneumonia, and
meningitis.
In enteric colibacillosis of piglets and calves the carcass appears
dehydrated but the intestine is flaccid and fluid-filled.
In calves, the abomasum is usually distended with fluid and may
contain a milk clot.
In both calves and pigs, the intestinal mucosae may appear normal
or hyperemic and there may be edema of the mesenteric lymph
nodes.
In calves affected with attaching and effacing E. coli there is
pseudomembranous ileitis as well as muco - hemorrhagic colitis 389 and
 Coli cont’d---
proctitis.
Diagnosis
Diagnose is done by complex method by considering
Epidemiology of the disease(age of animals)
Clinical signs
Necropsy findings
Laboratory methods
For laboratory confirmation the suitable specimens are
Faecal sample with enteric disease
Tissue specimens(spleen, liver with gall bladder and bone marrow)
are specimens of choice in septicemic form of colibacillosis.
The specimens are cultured on blood( selective media) and
MacConkey agars.
On blood agar the colonies are greyish, round and can be
haemolytic or not.
On MacConkey agar they form pinkish colony
Some of E. coli strains have a metallic sheen on EMB agar. 390
 Coli cont’d---
Test results are
In suspected cases of colisepticemia, isolation of E. coli in pure
culture from blood or from parenchymatous organs is considered
confirmatory.

391
 Coli cont’d---
However, when enterotoxigenic strains of E. coli are suspected, the
presence of either enterotoxins or fimbrial antigens can be
confirmed by immunological methods or molecular techniques such
as PCR.
Enterotoxins in the small intestine can be detected using methods
employing monoclonal antibodies.
Fimbrial antigens can be detected using ELISA or latex
agglutination.
The toxins produced by verotoxigenic and necrotoxigenic strains
can be detected by Vero cell assay.
Differential diagnosis
Colibacteriosis in calves and lambs must be differentiated from
Salmonellosis
Pasteurellosis
Rota and corona virus infections
They can be differentiated
392
 Coli cont’d---
By bacteriological and virological isolation and identification of
bacteria and virus and
 By considering the clinical signs and epidemiology of the disease.
Colibacillosis in piglets must be differentiated from
Swine pests
Erysipelas
Aujeszky's disease
Listeriosis
They can be differentiated by considering clinical signs,
epidemiology of the disease and off course by the result of
bacteriological and virological diagnosis in laboratory.
Treatment
Treatment of colibacteriosis must be done by complex method and
depends on types of colibacteriosis.
1. In coliform septicemia
Antimicrobials are given parenterally and may be given
393
 Coli cont’d---
continuously intravenously, more than once daily until recovery is
apparent.
Isolation of the organisms from blood and determination of drug
sensitivity is the ideal protocol.
Intravenous fluid and electrolyte therapy are administered
continuously until recovery is apparent.
2. In enteric colibacillosis
The considerations for treatment of enteric colibacillosis include
the following:-
Fluid and electrolyte replacement
 Antimicrobial and immunoglobulin therapy
 Antimotility drugs and intestinal protectants
Clinical management of outbreaks
The dehydration, acidosis and electrolyte imbalance are corrected
By the parenteral and oral use of simple or balanced electrolyte
solutions.
394
 Coli cont’d---
It can be administered through different routes and routes of
administration depends on degree of dehydration.
1. Calves with a history of acute diarrhea slightly dry oral mucosa,
good suck reflex, good muscle tone, alert, able to stand and warm
mouth.
These can be treated with oral fluids and electrolytes therapy.
 2. Calves with slight acidosis, weak suck reflex, good muscular
tone and warm mouth.
 Administer hypertonic saline (7.5% NaCl) intravenously at dose of
3 - 4 ml/kg BW
Administer oral fluids and electrolytes by stomach tube at 40- 60
ml/ kg BW and re-evaluate in 6 - 8 hours.
 3. Calves which are more dehydrated and have dry and cool oral
mucous membranes, recumbent, no suck reflex, and are very
depressed.
 Provide intravenous replacement and maintenance fluid and
electrolyte therapy for a period of 6-8 hours and up to 24-36 hours
395
 Coli cont’d---
if necessary.
Parenteral fluid composition which is a simple and effective
solution has
An equal mixture of isotonic saline (0.85%)
Isotonic sodium bicarbonate (1.3%) and
Isotonic dextrose (5%)
Some common electrolytes that can be administered parenterally
during acidosis and dehydration are
0.85% sodium chloride (isotonic saline)
1.3% sodium bicarbonate (isotonic)
1.3% sodium bicarbonate in 5% dextrose
5% sodium bicarbonate (hypertonic) during sever acidosis
Equal mixture of isotonic saline and isotonic sodium bicarbonate
Lactated Ringer’s solution
High-sodium, alkalinizing solution: Lactated Ringer’s solution plus
sodium bicarbonate (5 g/l) during acidosis and hyponatremia.
Calves with hypogammaglobulinemia can be given bovine gamma 396
 Coli cont’d---
globulin intravenously.
Enteric diseases may be treated by oral administration of
antimicrobial compounds which are active in GI tract.
The common and effective antibiotic drug that can be administered
per os during enteric colibacteriosis is
Amoxicillin
Dosage recommendations are
 Amoxicillin trihydrate (10 mg/kg every 12 h) for at least 3 days or
amoxicillin trihydrate - clavulanate potassium (12.5 mg combined
drug/kg every 12 h) for at least 3 days.
However, systemic infection require parenteral administration of
therapeutic agents and treatment should be based on antibiotic
susceptibility testing of isolates.
 Parenteral administration of broad-spectrum betalactam
antimicrobials (ceftiofur, 2.2 mg/kg intramuscularly or
subcutaneously every12 h). Or
397
 Amoxicillin or ampicillin, 10 mg/kg intramuscularly every 12 h) or
 Coli cont’d---
potentiated sulfonamides (25 mg/kg intravenously or
intramuscularly every 12 h) is recommended.
In calves with neonatal diarrhoea, milk should be with drawn and
replaced by fluids containing electrolytes and glucose.
The withholding of milk from diarrheic calves has been based on the
observations that lactose digestion is impaired.
So that resting of the intestine for a few days minimizes additional
osmotic diarrhea caused by fermentation of undigested lactose in
the large intestine.
Thus it has seemed logical not to feed the animal with milk which
must be digested.
But rather to provide readily absorbable substances such as glucose
- electrolyte mixtures orally.
In clinical management of outbreaks, the following principles
should be considered
 The veterinarian should visit the farm and conduct an
epidemiological investigation to identify risk factors.
 Examine each risk factor and how it can be minimized 398
 Coli cont’d---
Examine affected animals
 Identify and isolate all affected animals if possible
Treat all affected animals as necessary
Take laboratory specimens from affected and normal animals
 Make recommendations for the control of diarrhea in animals to be
born in the near future
Control and prevention measures
Effective control of colibacillosis can be accomplished by the
application of three principles:-
1. Reduction of the degree of exposure of the newborn to the
infectious agents
This predicate that newborn animals should receive ample amounts
of colostrum shortly after birth.
Because colostral antibodies can prevent colonization of the intestine
by pathogenic E. coli.
You know absorption of gamma globulin from the intestine declines
progressively after birth and is negligible by 36 hours. 399
 Coli cont’d---
2. Provision of maximum nonspecific resistance with
adequate colostrum and optimum animal management
This can be achieved by
 Supplying to neonates colostrum within 1 – 1: 30 hours and
Supplying clean, warm environment
For piglets new dietary regimes should be introduced
gradually at weaning time.
3. Increasing the specific resistance of the newborn by
vaccination of the dam or the newborn
This can be achieved by vaccination which includes: -
Commercially available killed vaccines containing prevalent
pathogenic E. coli serotypes to pregnant sows.
Vaccination of pregnant cows with purified E.coli K99
fimbrial or whole – cell preparation, often combined with
rotavirus antigen can be used to enhance colostral protection.
400
9. Salmonellosis
It is an infectious disease of many domestic animals including
poultry caused by bacteria genus Salmonella and is characterized by
Septicemia and enteritis in young animals
Latent infection in adult animals
As the result adult animals become carriers.
Salmonellosis is an economically important disease of all animal
species worldwide.
Salmonellosis and brucellosis have 4 points in common because
Both are important from the public health point of view
Both can lead to abortion
Both can lead to a carrier state likely to perpetuate the infection and
Both can lead to considerable financial loss to the farmers
Etiology
The genus Salmonella belongs to the family Enterobacteriaceae.
It is gram negative rod shaped or coccobacillary form bacteria.
They grow on conventional medium like blood agar and on selective
media for Enterobactericeae like XLD and brilliant green agar. 401
 Sal.cont’d---
Selective media for salmonella is Bismuth Sulfite Agar (BSA) and
has black colonies on BSA.
Currently there are 2463 serotypes (serovars) of Salmonella.
Serotyping of Salmonella is done based on somatic(O) and
flagellar(H) and occasionally capsular antigens(K).
By determining of the above antigens; there are two species of
Salmonella
S. enterica
S. bongori
S. enterica has 7 subspecies
 Salmonella enterica subsp. enterica
 Salmonella enterica subsp. arizonae
 Salmonella enterica subsp. bongori
 Salmonella enterica subsp. diarizonae
 Salmonella enterica subsp. indica
 Salmonella enterica subsp. salamae
 Salmonella enterica subsp. houtenae 402
 Sal.cont’d---
The majority of salmonellae of veterinary importance belongs to
Salmonella enterica subsp. enterica
The subspecies are further qualified by the serotype to give a final
designation such as S. enterica subspecies enterica serotype
typhimurium and written as S. typhimurium.
This nomenclature is now being used by the majority of
bacteriologist.
Salmonella can be classified into three main groups based on their
association with human and animal hosts: -
1. The first group consists of serotypes which have specificity to
the human host.
Salmonella typhi and Salmonella paratyphi.
 2. The second group contains serotypes adapted to specific animal
hosts.
Some of examples are
Salmonella dublin in cattle
 Salmonella abortusequi in horses 403
 Sal.cont’d---
Salmonella abortusovis in sheep and
Salmonella choleraesuis in pigs.
Salmonella pullorum and gallinarium in poultry
 3. The third group consists of un adapted serotypes which cause
disease in humans and animals
 Salmonella typhimurium is the best example of serotype in this
group.
Epidemiology
The epidemiology of salmonellosis is complex, which often makes
control of the disease difficult.
Salmonellosis occurs world wide and infect many mammals, birds
and reptiles.
The morbidity rate in outbreaks of salmonellosis in pigs, sheep, and
calves is usually high, often reaching 50% or more.
Morbidity and mortality are usually highest in calves under 12
weeks of age.
404
 Sal.cont’d---
The serotypes (serovars) that most commonly cause salmonellosis
in farm animal species are as follows
Spp of animals Serotypes or serovars
Cattle S. typhimurium, S. dublin and Salmonella newport
Sheep and goats S. abortusovis, S. typhimurium, S. dublin and Salmonella
anatum
Pigs S. typhimurium and S. choleraesuis
Horses S. typhimurium, S. abortusequi, S. anatum, S. newport, S.
enteritidis, Salmonella heidelberg, Salmonella arizona and
Salmonella angona.

405
 Sal. cont’d---
Salmonella serotypes of clinical importance and the consequence
of infection
Salmonella Hosts Consequence of infection
serotypes
Many animal species Enterocolitis
S. typhimurium
Human Food poisoning
Cattle Many disease conditions
S. dublin
Sheep, horses and dogs Enterocolitis and septicemia
S. choleraesuis Pigs Enterocolitis and septicemia
Poultry Often subclinical infection
S. enteritidis Many other species Clinical disease
Human Food poisoning
S. brandenburg Sheep Abortion

406
 Table cont’d---
Salmonella serotypes affecting only birds
S. pullorum Chicks Pullorum disease ( bacillary white diarrhoea)
S. gallinarum Adult birds Fowl typhoid
S. arizonae Turkeys Arizona or paracolon infection
Source of infection
Sick animals
Carrier animals

407
 Sal.cont’d---
They excrete the bacteria in faces.
Salmonella is spread by direct and indirect means.
Sick animals and carriers excrete and infect other animals directly,
or indirectly by contamination of the environment, primarily feed
and water supplies.
So that farm animals may be infected in many ways.
Some common routes of infection are
Animals -to- animal transmission, especially of the host-adapted
serovars
Contaminated animal feed and water ( ingestion)
Rarely through the mucosae of the upper respiratory tract and
conjunctiva
Mechanical vectors( insects) and rodents
Since salmonella is facultative intracellular organisms and can
survive in the phagolysosome of macrophages.
By this mechanism, it can evade the bactericidal effects of antibody
408
and complement.
 Sal.cont’d---
Persistence of infection in animals is the important epidemiological
features of salmonellosis.
Although most organisms are cleared from the tissues by the host
defense mechanisms, subclinical infection may persist with shedding
of small numbers of salmonella organism in the faces.
There is also latent infections in which salmonella is present in the
gall bladder but are not excreted.
So that clinical disease may develop from subclinical and latent
infections if affected animals are stressed.
Some of stress factors which may activate latent or subclinical
salmonellosis are
Intercurrent infections
Transportation
Overcrowding
Pregnancy
409
 Salm cont’d---
Extreme ambient temperature
Water deprivation
Oral antimicrobial therapy
Sudden changes in rations altering the intestinal flora
Surgical procedures requiring general anesthesia
Salmonellosis is a significant cause of economic loss in farm
animals because
Loss due to death because of salmonellosis
Economic loss for diagnostic purpose
Economic loss for control and eradication program(example costs of
cleaning and disinfection ).
Pathogenesis
Pathogenesis of salmonellosis is poorly understood.
But some of the general features associated with its virulence is 410
 Sal.cont’d---
known.
After oral infection with salmonellae, invasion of the host takes
place through intestinal wall in the terminal ileum and cecum and
progresses as far only as the mesenteric lymph nodes.
Progress beyond this point and development of the disease,
salmonellosis is determined by
Immune status and age of the host
Exposure to stress and
Virulence of the strain organism
The virulence of salmonella relates to
Their ability to adhere on host cell( the presence or absence of pilli)
Their ability to invade host cells(cytotoxin, endotoxin)
NB. LPS is responsible for endotoxin production and contribute to
the local inflammatory response which damages intestinal epithelial
cells and results in the development of diarrhoea.
Their ability to replicate in them and resist both digestion by
phagocytes and destruction by the complement components411of
 Sal. cont’d---
serum
Following adherence, probably through fimbrial attachment to the
surface of intestinal mucosal cells, the bacteria induce riffling of cell
membranes.
The ruffles facilitate the uptake of the bacteria in the membrane –
bound vesicles which often coalesce.
The organism replicate in this vesicles and are eventually released
from the cells which sustain only mild or transient damage of
intestinal mucousa and found in mesenteric lymph nodes.
In young animals and debilitated or aged are particularly susceptible
and the organism spread beyond the mesenteric lymph nodes occurs
And infection is established in liver cells, from this site the infection
invades the bloodstream and cause septicemic salmonellosis.
Systemic invasion may be sufficient to cause only bacteremia and
acute enteritis develops and abortion is the commons final sequel in
sheep and cattle.
Many animals survive this acute enteritis but localization of 412the
 Sal. cont’d---
and particularly gallbladder.
In healthy adults there may be no clinical illness when infection
first occurs but there may be localization of salmonellae in
abdominal viscera.
In the above both cases animals may become chronic carriers and
discharge salmonellae intermittently into feces and occasionally
into milk.
Clinical findings
Salmonellosis can be described as three syndromes classified
according to the severity of the disease as
1. Septicemia
2. Acute enterocolitis
 3. Chronic enterocolitis
 4. Abortion
 5. Terminal dry gangrene, osteitis and polyarthritis
1.Septicemic salmonellosis
Septicemic form of salmonellosis occurs in all age groups. 413
 Sal.cont’d---
However, it is most common in calves, in neonatal foals and in pigs
less than four months of age.
The onset of the disease is characterized by
Sudden high fever (40.5 - 420C)
Depression and recumbency
If treatment is delayed, many young animals with septicemic
salmonellosis die within 48 hours.
Surviving animals can develop persistent diarrhoea, arthritis,
meningitis or pneumonia.
In pigs with septicaemic salmonellosis there is a characteristic bluish
discoloration of the ears and snout.
2. Acute enterocolitis
Enterocolitis caused by salmonella organisms can affect most
species of farm animals irrespective of age.
Acute enterocolitis is characterized by
Fever
Depression 414
 Sal.cont’d---
Anorexia and profuse foul – smelling diarrhoea often containing
blood, mucous and epithelial casts.
Dehydration
Pregnant animals may abort
Severely affected young animals become recumbent and may die
within a few days of acquiring infection.
Onfarms with endemic salmonellosis, the milder clinical signs often
observed may be attributed to the influence of acquired immunity.
3. Chronic enterocolitis
It follows acute salmonellosis in pigs and occurs occasionally in
cattle and adult horses.
It is characterized by
Intermittent fever
Intermittent or persistent diarrhea with occasional passage of blood
mucous and firm fibrous casts
Gradual weight loss, leading to emaciation
415
 Sal. cont’d---
4. Abortion
It is a common manifestation of salmonellosis in cattle between
days of 124 and 270 of gestation.
When infection is associated with S. dublin, the organism
multiplies in the placenta, having been seeded there from a primary
lesion in other maternal tissues.
Cows that abort may have
 Fever and anorexia which results in hypogalactia
Some may have retention of fetal membranes.
In some cases, calves may be born shortly before term and die in
the perinatal period.
5. Terminal dry gangrene, osteitis and polyarthritis
Terminal dry gangrene due to endoarteritis of the extremities of the
limbs, ears, and tail may occur in calves with S. dublin infection.
Epiphyseal osteomyelitis affecting the metaphyses, and
polysynovitis and arthritis are also possible sequelae.
416
417
 Sal. cont’d---
Terminal dry gangrene of the extremities of calves is characterized
by
Lameness
 Swelling of the hind limbs below the fetlocks and separation of the
skin above the fetlock.
The distal portion of the limb is cool, not painful and the skin is dry
or moist.
There is a clear line of demarcation of the skin at the level of the
fetlock joints between the normal proximal skin and the distal
necrotic tissue.
The phalanges may be separated from the metatarsus.
The tips of the ears may be indurated and deviated medially and the
distal aspect of the tail may be dry and shriveled
Necropsy findings
The necropsy findings during salmonellosis depends on disease
syndromes.
418
 Sal. cont’d---
In septicemic form
Usually no gross lesions because of absence of time to develop as it
is peracute form.
However, sometimes there may be extensive submucosal and
subserosal petechial hemorrhages in different visceral organs.
In acute enterocolitis form
Gross lesions are most prominent in large and small intestines
There is inflammation as muco enteritis and hemorrhagic enteritis
Abomasitis with multiple mucosal erosions and petechiation
Intestinal contents are watery, have a putrid odor, contain mucous,
blood-tinged or whole blood
Mesenteric lymph nodes are enlarged
NB. In S. typhimurium infection, there is severe necrotic enteritis.
In chronic enterocolitis form
Discrete areas of necrosis on the wall of the cecum and colon with
thickened walls
419
Mesenteric lymph nodes and spleen are swollen
Diagnosis
Diagnosis of salmonellosis is done based on
Epidemiology( Age of affected animals and history of previous
outbreak of the disease)
Clinical picture of the disease
 Necropsy findings( enterocolitis with blood stained luminal content
and enlarged mesenteric lymph nodes)
Laboratory
Laboratory diagnosis is a confirmatory diagnose that can be done by
Isolation and identification of bacteria
 Serotyping of identified bacteria by slide agglutination test
 Serological tests like ELISA and agglutination techniques used to
know the past infection and to identify latent infection
Rising of antibody titer using paired serum samples is indicative of
active infection.
Molecular techniques like DNA probes can be used to screen large
numbers of faecal samples for salmonella.
420
Differential diagnosis
The septicemic form of salmonellosis in calves must be
differentiated from
 Coliform septicemia and can be differentiated by bacteriological
examination of blood, feces, and tissues.
How ever, salmonellosis occurs most commonly during the second
and third weeks of life in contrast to coliform septicemia which
occurs most commonly in the first few days of life.
Acute enterocolitis salmonellosis must be differentiated from
Coccidiosis
Coccidiosis occurs most commonly in young cattle 2 - 8 months of
age
Chronic enterocolitis salmonellosis must be differentiated from
1. Johne's disease or chronic molybdenum poisoning
However, dysentery and epithelial casts do not occur in these
diseases but present in salmonellosis. 421
 Sal. cont’d---
2. Massive stomach fluke infestations may also cause diarrhea and
dysentery.
Other diagnoses that must be considered include:
Clostridiosis due to Clostridium perfringens type A and Clostridium
difficile may result in peracute hemorrhagic diarrhea, marked
dehydration and rapid death.
Chronic diarrhea due to salmonellosis may resemble parasitism,
granulomatous enteritis or lymph sarcoma.
In pigs septicemic salmonellosis must be differentiated from
 1. Hog cholera
2. African Swine fever(ASF)
3. Coliform gastroenteritis( it occurs in recently weaned piglets)
while salmonellosis occurs in 1 – 4 months of age
4. Pasteurellosis mostly has respiratory form with atrophic rhinitis
 5. Acute erysipelas which is characterized by typical skin lesions,
fever and swollen joints.
422
 Sal. cont’d---
Treatment
Treatment of salmonellosis can be done using
1. Primary therapy using antimicrobial drugs
2. Supportive therapy
1. Primary therapy using antimicrobial drugs
Antibiotic therapy should be based on the result of antibiotic
susceptibility test
Because R- plasmids coding for multiple resistance are
comparatively common in salmonella.
Oral antimicrobial therapy should be used judiciously for treating
enteric salmonellosis.
Because it may disturb the normal intestinal flora, extend the
duration of salmonella excretion and increase the probability of
drug resistance development.
The most common drugs used to treat salmonellosis in ruminants
are
In the septicemic form of the disease, intravenous antibiotic therapy
must be used.
423
Some common antimicrobial drugs that are used to treat
 Sal. cont’d---
salmonellosis are
 Ceftiofur 5 mg/kg BW intramuscularly/ 24 hours for 5 days is
effective
 In calves with S. dublin infections trimethoprim-sulfadoxine is
recommended, given parenterally daily until clinical recovery
occurs.
Ampicillin and amoxicillin are also effective
 Gentamycin at 3 mg/kg BW combined with ampicillin at 20 mg/kg
BW given intravenously at S-12-hour intervals is recommended.
 An alternative is trimethoprim-sulfonamide given twice daily
intravenously at a combined dose of 30 mg/kg BW.
Sulfadiazine, sulfadoxine and sulfamethoxazole are the best
sulfonamides to combine with trimethoprim for salmonellosis in the
horse.
Orally of mass medication of the water supply with chlortetracycline
and sulphamethazine can be used to treat enterocolitis form of
salmonellosis 424
2. Supportive therapy
Fluid and electrolyte replacement therapy is required to solve
dehydration, acidosis and shock.
It is possible to administer
0.85% sodium chloride (isotonic saline)
1.3% sodium bicarbonate (isotonic)
1.3% sodium bicarbonate in 5% dextrose
5% sodium bicarbonate (hypertonic) during sever acidosis
Equal mixture of isotonic saline and isotonic sodium bicarbonate
Lactated Ringer’s solution
High-sodium, alkalinizing solution: Lactated Ringer’s solution plus
sodium bicarbonate (5 g/l) during acidosis and hyponatremia.
Prevention and control measures
1. If the farm is free from salmonellosis
Control is based on reducing the risk of exposure to infection.
This can be achieved by
Prevention of introduction of infection by having strong
“biosecurity measures” in your farm that can be achieved by 425
 Sal. cont’d---
A closed – herd policy should be implemented when feasible
Animals should be purchased from reliable sources and remain
isolated until negative for salmonella on three consecutive tests.
 If possible, purchase animals when they are older, to provide an
opportunity for specific and nonspecific immunity to develop.
Purchase animals from vaccinated herds are desirable
Introduce only those animals likely not to be carriers
Have prophylactic disinfection schedule in your farm
Have prophylactic vaccination program for your herd
2. If the farm is not free from salmonellosis
Limitation of the spread of the agent within the herd is essential
which can be achieved by
Identify carrier animals and either cull them or isolate and treat them
vigorously.
Treated animals should be re - sampled subsequently to determine 426
 Sal. cont’d---
whether a ' clean' status has been achieved
The prophylactic use of antimicrobials such as oxytetracycline in
the feed at the rate of 10 gm/ tone of feed or chlortetracycline in the
drinking water at the rate of 55 mg/liter is crucial
Restrict the movement of animals around the farm
 The water supply should be provided in troughs that are not
accessible to fecal contamination.
 Rigorous disinfection of buildings is important after every
outbreaks
 An all-in and all-out policy should be adopted and steam cleaning
and chemical sterilization performed after each batch of animals is
essential
 Suitable construction of housing is important
Heat treatment of feed is an effective procedure for pigs.
 Disposal of infective material should be done with care. For
example carcasses should be burned
All persons working on infected premises should be warned of the
427
hazards to their own health.
Diseases associated with Actinomyces, Actinobacillus and
Dermatophilus spp .
Here we will see
10. Actinomycosis
 11. Actinobacillosis
12. Dermatophilosis
10. Actinomycosis(Lumpy jaw)
Actinomycosis is a chronic infectious disease of cattle.
It is characterized by osteomyelitis of the bones of head, particularly
the mandible and maxilla and suppurative lesions in connective
tissue and bones.
Sometimes soft tissues of head and neck are involved in the form of
granulomas.
This leads eventually to replacement of normal bone by porous bone
that laid down irregularly and honeycombed with sinus tracts
containing pus.
Etiology
428
 Act. cont’d---
It is caused by Actinomyces bovis .
However, occasionally Corynebacterium pyogenes and Staph.
aureus have been found in association with Act. bovis.
Act. bovis is a gram positive filamentous bacteria.
Rarely they tend to be a mixture of cocci, rods and pear shaped
cells.
Occasionally short branching forms may be seen.
It grows on conventional media like blood agar very well in
capnophilic condition(5 – 10% Co2 ).
Epidemiology
The disease is sporadic and common in cattle.
Occasional cases occur in pigs and horses and rarely in goats.
Although actinomycosis occurs only sporadically, it is important
disease because of
Its widespread occurrence and poor response to treatment.
Actinomyces bovis is a common inhabitant of the bovine tooth.
Infection is presumed to occur 429
Gram positive branching (filamentous) Actinomyces

430
 Act. cont’d---
Through wounds to the buccal mucosa caused by sharp pieces of
feed or foreign material
 Through dental alveoli and may account for the more common
occurrence of the disease in young cattle when the teeth are
erupting.
Rarely infection of the alimentary tract wall is probably related to
laceration by sharp foreign bodies.
Source of infection and transmission
Most actinomycotic infections are endogenous infections
Infection is caused by introduction of a commensals strain into
susceptible tissues of its host.
Rarely biting is another means of transmission.
Pathogenesis
A. bovis has an access to the alveolar region of the jaw in cattle
from the oral cavity by invasion of mandible and maxillary bones.
It reaches to alveolar region of the jaw due to the presence of
different trauma on the mucosa by rough feed, tooth eruption etc.431
 Act. cont’d---
As the result the bacteria initiates to rarefying osteomyelitis and soft
tissue reaction.
This condition being referred “ Lumpy jaw” in cattle.
Lumpy jaw refers abscess( abscesses) that grow on the head and
neck of the infected animal.
The bacterial colonies form in affected tissues with clubs of
mineralized calcium phosphate forming around them to create
microscopic “ club colonies ”or rosettes.
The club formation is due to phosphatase reaction of the bacteria and
host reaction to chronic infection.
Granulation, mononuclear infiltration and fibrosis occur in the
lesions with sinus tracts to out side.
So the exudate will come out through tracts and contain pus.
The pus contains “sulphur granules” that are about 1 – 2mm in
diameter
The sulphur granules contain club colonies of A. bovis that can be
examined microscopically after breaking of the sulphur granules.432
Microscopic “ club colonies ”of Actinomyces bovis

433
Cows suffering from lumpy jaw

434
435
436
Drainage of exudate through out tracts and contain pus.

437
The bone of the mandible is dissolved and the bone trying to
repair itself, leaving a ‘honeycomb’ effect

438
Clinical signs
Actinomycosis usually occurs in chronic form.
It has two forms
1. Actinomycosis with involvement of hard tissues( bone and teeth)
2. Actinomycosis with involvement of soft tissues
1. Actinomycosis with involvement of hard tissues( bone and
teeth)
Actinomycosis of the jaw commences as a painless, bony swelling
which appears on the mandible or maxilla, usually at the level of the
central molar teeth.
The enlargement may be diffuse or discrete and in the case of the
mandible may appear only as a thickening of the lower edge of the
bone with most of the enlargement in the inter madibular space.
The more common, discrete lesions on the lateral surfaces of the
bones are more readily observed.
Some lesions enlarge rapidly within a few weeks, others slowly
over a period of months.
439
 Act. cont’d---
The swellings are very hard, immovable and in the later stages,
painful to the touch.
They usually break through the skin and discharge through one or
more openings.
Teeth embedded in the affected bone become malaligned(imperfect
aligned) and painful and cause difficult mastication with
consequent loss of body condition.
2. Actinomycosis with involvement of soft tissues
It occurs rarely.
The most common form of actinomycosis of soft tissues is
involvement of the esophageal groove region, with spread to the
lower esophagus and the anterior wall of the reticulum.
The syndrome is one of impaired digestion.
Less common lesions of soft tissue include orchitis in bulls and the
trachea causing partial obstruction, and abscess in the brain or
lungs.
440
Necropsy findings
Rarefaction of the bone and the presence of loculi( cavity) and
sinuses containing thin, whey-like pus with small, gritty granules is
usual.
The presence of “club colonies” containing the typical, thread -like
bacteria is characteristic of the disease.
These formations may be seen on microscopic examination of
smears made from crushed granules in pus or on histological
examination of section.
Granulomatous lesions containing pockets of pus may be found in
the esophageal groove, the lower esophagus and the anterior wall
of the reticulum.
Spread from these lesions may cause a chronic, local peritonitis.
Diagnosis
It is mostly depends on
Clinical signs and demonstration of the organism with sulphur
granules either by direct microscopy or after gram staining.
441
 Act. cont’d---
Observation of club colony of A. bovis in histopathological tissues
after Eosin Hematoxylin staining.
Isolation and identification of the bacteria
Direct microscopy
Sulphur granules are the best specimens for direct examination in
infections caused by A. bovis.
Procedure
Put the pus or exudate in a Petri dish.
Wash the pus with a little distilled water to expose the yellowish
sulphur granules of A. bovis
Then put the granules on slide having a drop of 10% KOH .
Gently crushed the granule by applying a pressure on cover slip
Look the crushed granule under low power dry objective of
microscope.
So you will see the clubs of Actinomyces bovis.
Or you can prepare a smear from granules.
442
Stain the smear by Gram.
 Act. cont’d---
Look under immersion oil.
You will see Gram – positive, branching filaments.
Rarely short filaments, pleomorphic may predominate or they tend
to be a mixture of cocci, rods and pear shaped cells.
Sulphur granules in pus from A. bovis infection in cattle

443
Branching filament gram positive bacteria of A. bovis

444
Club colonies in a tissue section from A. bovis infection in cows
with lumpy jaw

445
Differential diagnosis
Lumpy jaw must be differentiated from
1. Abscesses of the cheek muscles and throat region
Abscesses of the cheek muscles and throat region are
characterized by their
Movability and localization in soft tissues compared to the
immovability of an actinomycotic lesion .
Pus may be thin, fetid, or caseous depending on the duration of the
abscess.
Prompt recovery follows opening and drainage
2. Foreign bodies or accumulations of dry feed jammed between
the teeth and cheek
It commonly causes a clinical picture which resembles that
associated with actinomycosis
So that the mouth cavity should be inspected if the enlargement
has occurred suddenly.
446
 Act. cont’d---
3.Tumor
4. Tuberculosis
5. Actinobacillosis (wooden tongue)
Treatment
Treatment is with surgical debridement and antibacterial therapy,
particularly Iodides is effective.
In bovine actinomycosis , iodine compounds given orally( 4 to 8
gm) daily or intravenously(75mg/ kg) weekly are used.
Iv treatment with iodine must be interrupted when signs of toxicity
appear(hyper salivation, anorexia and vomiting) but you can restart
again after some weeks again.
Accessible soft tissue lesions can be drained or excised.
Additional treatment recorded as being effective includes isoniazid
given orally at the rate of 10-20 mg/kg body weight daily for about
30 days.
NB. Bacteriologic cure of lumpy jaw will not restore the normal
bone structure. 447
 Act. cont’d---
But the process can be arrested by systemic medication aided by
drainage, lavages( iodine) and debridement of lesions.
Prevention and control measures
Avoid feeding of animals that are strong and rough.
Disposal of animals with discharging lesions is important, although
the disease does not spread readily unless predisposing
environmental factors cause a high incidence of oral lacerations.
11. Actinobacillosis (wooden tongue)
Actinobacillosis is an infectious disease of cattle, sheep and pigs.
In cattle actinobacillosis is characterized by
 Formation of hard immoveable tongue called wooden tongue
(timber tongue).
Pyogranulomatous lesion around the head, neck and limbs.
Occasionally pyogranulomatous lesions in the internal organs.
In sheep actinobacillosis is characterized by
 Pyogranulomatous lesions usually in head and neck region.
In pigs it is characterized by 448
 Actinobac. cont’d---
Rare granulomatous abscesses in mammary glands
Etiology
Actinobacillosis is caused by Actinobacillus lignieresii.
Actinobacillus lignieresii is gram – negative , medium – sized rods.
Rarely they can have coccal forms.
They are non – motile, non – spore forming and non – acid fast.
Most species of Actinobacillus grow on sheep or ox blood agar.
The colonies are small , glistening and non – haemolytic.
A. lignieresii grow on MacConkey agar.
The colonies are first pale but become pinkish as A. lignieresii is
late lactose fermenter.
If the Gram – stained is made from the crushed granules, the
presence of medium sized Gram – negative rods would suggest
actinobacillosis rather than the filamentous Gram positive
actinomycosis.
A. lignieresii may be recovered in pure culture from the lesions.
However, other pyogenic organisms may also be present. 449
Actinobacillus lignieresii in Gram staining from pure culture

450
Gram negative bacillus bacteria of A . lignieresii

451
Club colonies in a tissue section from Actinobacillus lignieresii
infection in cows wooden tongue

452
Epidemiology
The disease has world wide occurrence.
It occurs sporadically but very rarely in epidemics.
It affects mostly cattle, sheep and rarely horses and pigs.
Actinobacillus lignieresii is a normal inhabitant of the oral cavity
and rumen of ruminants.
Infection in soft tissue results from damage to the oral mucosa.
In cattle, infection most commonly occurs through ulcerating or
penetrating lesions to the sulcus of the tongue, penetrating lesions in
the apex, and lacerations to the side of the body of the tongue caused
by the teeth.
In sheep, the different nature of prehension of food leads to lesions
predominantly in the lips and cheeks with occasional extension to
the mucous membranes of the turbinate and the soft tissue of the
head and neck.
Risk factors
The risk factors for occurrence of wooden tongue are
453
 Actinobac. cont’d---
Abrasive pasture species or pastures with spiny awns.
Traumatic lesions caused by nose grips or jugular venipuncture.
Pathogenesis
Localization of the organism is followed by an acute inflammatory
reaction.
This results in the development of granulomatous lesion in which
necrosis and suppuration occur.
 Finally this lesion will discharge pus to the exterior and spread to
regional lymph nodes is usual.
Lingual (tongue) involvement in cattle causes interference with
prehension and mastication due to acute inflammation in the early
stage and distortion of the tongue in the later stages.
Visceral involvement is recorded and is identical with that described
under actinomycosis.
Clinical findings
Cattle 454
 Actinobac. cont’d---
In cattle, there are three types of actinobacillosis
 1. Glossal actinobacillosis
 2. Lymphadenitis form of actinobacillosis
 3. Cutaneous actinobacillosis
1. Glossal actinobacillosis
 Onset of glossal actinobacillosis is acute, affected animals being
unable to eat for about 48 hours.
Excessive salivation and gentle chewing of the tongue is common
On examination the tongue is swollen and hard, particularly at the
base, the tip appearing normal.
Manipulation of the tongue causes pain and resentment.
Nodules and ulcers are seen on the sides the tongue.
In the later stages, acute inflammation is replaced by fibrous tissue
and the tongue becomes shrunken and immobile which causes
considerable interference in prehension.
Visible and palpable enlargement of submaxillary and parotid
lymph nodes is common. 455
Cattle affected by glossal form of actinobacillosis

456
457
 Actinobac. cont’d---
The photo shows a tongue removed from a cow with wooden tongue.
Note the yellow lumps of 'actino' granules bursting out of the
swollen tongue — both in the black pigmented section and lower down
near the base.

458
 Actinobac. cont’d---
2. Lymphadenitis caused by actinobacillosis
Lymphadenitis is common and is often independent of lesions in the
tongue.
There may be visible and palpable enlargement of the submaxillary
and parotid nodes.
Local firm swellings develop and often rupture with the discharge
of thin, non -odorous pus.
Healing is slow and relapse is common.
Enlargement of the retropharyngeal nodes causes interference with
swallowing and loud snoring respiration.
3. Cutaneous actinobacillosis
Cutaneous actinobacillosis is also recorded with actinobacillosis
granulomas .
It occurs as atypical but visible in areas such as external nares,
cheeks, skin or eyelid and hind limbs.
External trauma from abrasive materials in the environment is the
usual initiating cause. 459
 Actinobac. cont’d---
Lesions are several centimeters in diameter and are pliable or firm
and painful on palpation, red, and can bleed easily.
Lymphadenitis form of actinobacillosis

460
Cutaneous form of actinobacillosis

461
Cutaneous form of actinobacillosis

462
Cutaneous form of actinobacillosis which may cause mastitis

463
 Actinobac. cont’d---
Actinobacillosis in sheep
In sheep, glossal lesions are not usual.
Lesions (up to 8 cm in diameter) occur in the lower jaw, face and
nose or in ventral side of the neck.
It may extend to the cranial or cervical lymph nodes.
The lesions may discharge yellow-green pus containing granules
through a number of small openings.
Extension of lesions with formation of fibrous tissues may
physically impede prehension and respiration.
Thick scabby lesion on the lips may cause difficulty in eating
resulted in death of starvation.
Lesions also extend to the nose causing persistent bilateral nasal
discharges.
Actinobacillus lignieresii is also an occasional cause of mastitis in
ewes and cows.
A similar involvement of the lips with abscessation in the area of the
madibular lymph nodes is recorded in camels. 464
 Actinobac. cont’d---
In horses the disease is uncommon.
However, affected horses may show
Interamandibullar phlegmon or
 Infection of the tongue or of the muzzle
 Infection at other body sites.
Necropsy findings
Necropsy examination is not usually carried out in cattle affected
by the disease.
In sheep, lymphangitis and abscesses containing thick, tenacious,
yellow-green pus occur around the local lesion.
Typical club colonies are visible on staining sections of affected
tissue.
Culture of material from lesions usually detects the presence of
Actinobacillus lignieresii.
Diagnosis
Clinical signs and necropsy findings helps to suspect
actinobacillosis. 465
 Actinobac. cont’d---
Although, microscopically there is observation of " sulfur
granules” consist of club like rosettes with a center mass of bacteria,
it is not the pathognomonic to Actinobacillus lignieresii.
So, examination of smear or culture of pus for the presence of the
organism is advisable.
Club colonies of Actinobacillus lignieresii in histopathological
sections stained in H and E stain ×400 and gram staining

466
 Actinobac. cont’d---
Differential diagnosis
Actinobacillosis must be differentiated from
 Foreign bodies in the mouth
Early stage of rabies
 Esophageal obstruction
 Tuberculosis
Treatment
It is important to treat the affected animals quickly.
The standard treatment is iodides with good and permanent
response through reduction of the severity of the fibrous tissue
reaction.
Potassium iodide(KI) is administered orally in cattle (6 - 10 g/day
for 7 - 10 days) until signs of iodism appear (lacrimation, anorexia,
coughing and dandruff).
Sodium iodide is administered IV (1g/12 kg BW) as a 10% 467
468
 Actinobac. cont’d---
solution) in one dose to both cattle and sheep, acute signs disappear
in 1- 2 days after treatment.
The sulfonamides, penicillin, streptomycin, and the broad-
spectrum antibiotics are also used.
Streptomycin, given by intramuscular injection (5 g/day) for 3
days repeated if necessary.
They give good result in cattle when combined with iodides and
local surgical treatment.
Isoniazid has been reported on favorably as an adjunct to antibiotic
or iodide therapy in cattle.
The daily dose rate recommended is 10 mg/kg body weight orally
or intramuscularly, continued for 3 - 4 weeks.
Cutaneous actinobacillosis may require an extended course of
treatment with streptomycin and/or dihydrostreptomycin for 2 - 4
weeks to achieve resolution.
Prevention and control measures
Restriction of the spread of disease is best implemented by quick469
 Actinobac. cont’d---
Quick treatment of affected animals and
Prevention of contamination of pasture and feed troughs.
Isolation or disposal of animals with discharging lesions is essential
although the disease does not spread readily unless predisposing
environmental factors cause a high incidence of oral or skin
lacerations.
12. Dermatophilosis (mycotic dermatitis, cutaneous
streptotrichosis, lumpy wool of sheep)
The disease in sheep is commonly called mycotic dermatitis or
lumpy wool of sheep.
In cattle it is called cutaneous streptotrichosis.
Dermatophilosis (also known as streptothricosis in cattle or in
sheep as ‘lumpy wool disease’) is an exudative, pustular dermatitis
that affects mainly cattle, sheep and horses.
It also affects goats, dogs and cats, many wild mammals, reptiles
and occasionally humans.
470
 Derm. cont’d---
So that it has zoonotic implication.
It has economic impact in tropical and sub - tropical countries
because
In sheep dermatophilosis damage to the fleece and causes severe
losses up to 30% loss of value of wool and 40% loss of skin value.
Other losses in sheep are caused by interference with shearing and a
very great increase in susceptibility to blowfly infestation.
In cattle losses are from
Direct animal loss due to death
 Decreased work ability of affected oxen
 Reproductive failure from vulval infection or infection on the limbs
of males preventing mounting
Death from starvation of calves of dams with udder infection
 Loss of animal meat and milk production and downgrading of hides.
That is why, it is the four major bacteriological diseases with
equivalent importance to contagious bovine plueropneumonia and
brucellosis in tropical countries. 471
Etiology
The infective agent is Dermatophilus congolensis.
The bacteria require damage to the skin from other causes to
establish infection.
D. congolensis is filamentous and branching bacteria.
Stain either by Gram or Giemsa staining.
The better one is Giemsa staining because it shows the morphology
of bacterium clearly.
You can use Gram – staining but the bacterium and debris may
seem to absorb the crystal – violet iodine complex and become
dark.
To solve this problem you must modify the Gram staining method
by leaving the crystal violet on smear for 2 – 3 seconds.
After this the morphology of the bacterium is easier to see.
The organism is dimorphic and grows as branched filamentous
mycelia containing dormant zoospores.
Zoospores are transformed by moisture to the infective
stage of motile isolated cocci. 472
 Der. cont’d---
Mature filaments form of Dermatophilus congolensis is
composed of
Motile coccal zoospores(1 µm in diameter) in parallel lines
 At least two abreast, resulting in a “ tram – track “ appearance.
If the flakes of the scab are treated roughly when the smears are
made, the filaments will disintegrate and only Gram positive cocci
zoospores) will be seen.
Since the morphology of D. congolensis is unique, strong
presumptive diagnosis of streptothricosis ( dermatophilosis) can be
made on direct examination of stained smears alone.
Although isolation of D. congolensis is not necessary for diagnosis
of streptothricosis, it necessary to culture it in laboratory.
Because it grows on conventional media like sheep or ox blood
agar under 5 – 10 % Co2.
Development cycle of D. congolensis
The free motile zoospore starts to grow and form germ tube.
473
The germ tube starts to form horizontal septa of immature
D. congolensis in Giemsa staining

474
D. congolensis in gram staining

475
 Derm. cont’d---
filament.
Then transverse and longitudinal septa are formed.
 The transverse and longitudinal septa formation leads to binary
fission and formation of cocci.
 When they matured, these cocci will have polar flagella then they
are called zoospores.
 Zoospores are motile by polar flagella and are infective.
 Zoospores can initiate an infection in macerated or traumatized
skin.
Development cycle of D. congolensis

476
 Der. cont’d---
Epidemiology
The disease occurs in all areas of the world.
It occurs as epizootic in tropical and subtropical areas of the world
where it causes considerable economic loss.
In temperate climates the occurrence of the disease is usually
sporadic.
However, it can still be of considerable economic importance where
predisposing factors pertain there.
The disease affects cattle, sheep, goats, horses, donkeys and
occasionally in deer, pigs, camels and wildlife species.
Animals of all ages groups are susceptible including suckling with
a few weeks old.
Source of infection
The major source of infection are
 Clinically diseased animals with minor active lesions on the face
and feet
 Healthy carrier animals with infection in scabs still carried in the
477
 Derm. cont’d---
hair and wool from healed lesions.
D. congolensis is not highly invasive and does not normally breach
the barriers of healthy skin.
These barriers include
 The stratum corneum
The superficial wax layer produced by the sebaceous glands
The physical barrier of the wool.
However on the feet and face these barriers are easily and
commonly broken by abrasive terrain or thorny and spiny forage and
feedstuffs
Dermatophilus may infect these lesions and may be transmitted
mechanically by feeding flies to result in minor infection on the face
and feet.
Transmission
Transmission occurs from the carriage lesions by
 Contact from the face of one animal to the fleece or skin of another
478
Contact from the feet to the skin during mounting.
 Derm. cont’d---
Infection can be transmitted mechanically by flies and ticks
 Mediate dermatophilosis infection can be transmitted by
contaminated dips.
Risk factors for occurrence of dermatophilosis
There are many risk factors but the commons are
1. Environmental and management risk factors
 2. Host risk factors
 3. Pathogen risk factors
1. Environmental and management risk factors
Prolonged wetting of the fleece is the major risk factor and leads to
emulsification of the wax barrier and maceration of the skin surface
with disruption of the stratum corneum of skin in sheep.
Some of the examples are
 1. A prolonged and heavy rain is sufficient to do this especially if
followed by warm and humid weather that retards drying of the
fleece.
 2. Dipping, jetting, or spraying of sheep for external parasites 479
 Derm. cont’d---
Shearing cuts also destroy the barriers of the skin and cuts may
become infected mechanically by flies, physically by tight
yarding after shearing, and by mediate infection in dips when
sheep are dipped immediately following shearing.
Skin infection can also occur following infection with
contagious ecthyma.
In cattle, climate is also one of environmental risk factors.
That is why dermatophilosis occurs sporadically in temperate
zones.
Sporadically occurrence of dermatophilosis is associated with
abrasions caused by mounting.
The use of periodic showers or continual misting to cool cattle
during hot periods
Intercurrent disease and stress are also risk factors and in
infected dairy herds. 480
 Derm. cont’d---
As in sheep, the disease in cattle requires disruption of natural skin
barriers.
However, prolonged wetting of the skin of cattle does not appear to
be a major predisposing factor by itself.
So that the seasonal occurrence of dermatophilosis is associated
with a concomitant increase in tick and insect infestation.
For example, a recent study in Ethiopia found that although
prevalence was higher in cattle in the wet season.
However, it was associated with infestation of A. variegatum.
Tick infestation, particularly with
Ambylomma variegatum
 Hyalomma asticum
 Boophilus microplus
Infestation is strongly associated with the occurrence of extensive
lesions of dermatophilosis which can be minimized by the use of
acaricides.
481
 Derm. cont’d---
The lesions of dermatophilosis on the body do not occur at the
predilection sites for ticks and it is thought that the importance of
tick infestation relates to a tick-produced immune suppression in the
host rather than mechanical or biological transmission.
Lesions do occur at predilection sites for biting insects, mainly
Stomoxys spp.
 Lyperosia spp.
 Glossinia spp.
Calliphoria spp. and mosquitoes.
In Africa the disease is often combined with demodicosis to
produce a more severe and often fatal combination.
Trauma to the skin produced by thorny bushes and the Ox-pecker
bird (Buphagus africanus africanus) can also initiate lesions.
Horses biting flies (Stomoxys calcitrans) are thought to act as
mechanical vectors of the infection and the house fly (Musca
domestica) can carry infection.
482
 Derm. cont’d---
2. Host risk factors
There are breed differences in susceptibility in cattle and sheep.
In Africa, some native cattle breeds( N'dama and Muturu cattle
breeds) and native sheep are resistant.
While Zebu, white Fulani and European breeds are susceptible.
In the Merino, sheep of the strong or medium wool strains are more
susceptible.
Open-fleeced sheep, and sheep with a low-wax and high-suint
content in their fleece are more prone to infection.
3. Pathogen risk factors
D. congolensis does not live well off the body and in the normal
environment, and is susceptible to the external influences of PH and
moisture fluctuations.
In dry scab material the bacteria can exist for least 13 years kept at
room temperature
483
Pathogenesis
The natural skin and wool waxes act as effective barriers to
infection.
Minor trauma or maceration by prolonged wetting allows
establishment of infection and multiplication of the organism in the
epidermis.
The formation of the typical pyramidal-shaped crusts is caused by
repeated cycles of invasion into the epidermis by hyphae, bacterial
multiplication in the epidermis, rapid infiltration of neutrophils and
regeneration of the epidermis.
In sheep, the extensive maceration of skin that can occur with
prolonged fleece wetting can result in extensive skin lesions under
the fleece.
In cattle, tick infestation suppresses immune function and promotes
the spread of the lesion.
Secondary bacterial invasion may occur and gives rise to extensive
suppuration and severe toxemia.
484
 Derm. cont’d---
The pathogenic factors are very diverse.
However, the most important one is the enzymes (adenase and
lecithinase) which are produced by D. congolensis that have a
keratinolytic nature on epidermis.
Clinical findings
Lesions in sheep are commonly not visible.
Because they are obscured by the fleece but the crusts can be
palpated as hard masses at the surface of the skin (lumpy wool
disease).
Typically they are distributed irregularly over the dorsal midline
with 'ribs' spreading laterally and ventrally.
The crusts are roughly circular and thick up to 3 cm, often distinctly
pyramidal with a concave base, often pigmented and the underlying
skin is moist and reddened.
The muzzle, face and ears and the scrotum of rams, may also be
involved.
485
 Derm. cont’d---
Dermatophilosis in sheep

486
Dermatophilosis in sheep

487
Dermatophilosis in goats

488
 Derm. cont’d---
Dermatophilosis in cattle is characterized by
Pustules in the early stage of infection and the hair over the infected
site is erect and matted in tufts (paintbrush lesions) with greasy
exudate forming crumbly crusts which are hard to remove.
These develop to scabs which are greasy and fissure at flexion
points and finally the scabs become hard, horny and confluent.
 The scabs vary in color from cream to brown and are 2-5 cm in
diameter and are often in such close apposition that they give the
appearance of a mosaic.
 In the early stages the crusts are very tenacious and attempts to lift
them cause pain.
 Beneath the crusts there is granulation tissue and some pus.
 Lesions usually occur on the neck, body and the back of the udder
and may extend over the sides and down the legs and the ventral
surface of the body 489
Dermatophilosis in cattle

490
Dermatophilosis in cattle

491
Streptothricosis in cow

492
 Derm. cont’d---
In horses dermatophilosis is characterized by
Lesions in horses are similar to those in cattle.
 The hairs are matted together over the lesion and an exudative
dermatitis produces a firm mat of hairs and debris just above the
skin surface.
 If this hair is plucked the entire structure may lift off, leaving a
characteristic ovoid, slightly bleeding skin area.
No pruritus or irritation is apparent although the sores are tender to
the touch.

493
Dermatophilosis in horses

494
495
496
Necropsy findings
In animals that die due to dermatophilosis there is
Extensive dermatitis
 Sometimes secondary bacterial complication cause pneumonia and
often evidence of inter current disease.
Diagnosis
Diagnose of dermatophilosis is done based on the epidemiology,
clinical and laboratory diagnosis.
The causative organism may be isolated from scrapings or a biopsy
section and is much easier to isolate from an acute case than a
chronic one.
Typical branching organisms with double rows of zoospores can be
seen in a stained impression smear ( Giemsa or gram staining)made
directly from the ventral surface of a thick scab pressed firmly onto
a slide.
497
Laboratory diagnosis of Dermatophilus congolensis
Specimens
A tuft of hair that is plucked from the lesions detaches with scab
materials adhering to it is a specimen to isolate D. congolensis.
Direct microscopy
Prepare the smear by shaving small pieces of materials from scabs
with scalpel.
This flakes of scabs are softened with a few drops of distilled water
on microscope slide.
Smear is prepared by leaving a few flakes of scab material intact on
slide.
Dry the smear with air
Fix it with methanol
Stain either by Gram or Giemsa staining.
 The better one is Giemsa staining because it shows the morphology
of bacterium clearly.
You can use Gram – staining but the bacterium and debris may seem
to absorb the crystal – violet iodine complex and become dark.
498
A tuft of hair being plucked from a horse with streptothricosis

499
 Derm. cont’d---
 To solve this problem you must modify the Gram staining method
by leaving the crystal violet on smear for 2 – 3 seconds.
After this the morphology of the bacterium is easier to see.
Since the morphology of D. congolensis is unique, strong
presumptive diagnosis of dermatophilosis can be made on direct
examination of stained smears alone.
D. congolensis is filamentous and branching gram positive
bacterium if it is stained by Gram.
Mature filaments are composed of
 Motile coccal zoospores(1 µm in diameter) that are arranged in
parallel lines
 At least two abreast, resulting in a “ tram – track “ appearance
If the flakes of the scab is treated roughly when the smears are
made, the filaments will disintegrate and only Gram positive cocci
(zoospores) will be seen.
The organism can also be demonstrated by Direct Fluorescent
Antibody Test(FAT). 500
 Derm. cont’d---
ELISA has also been used to detect serological evidence of
infection with D. congolensis .
D. conglesense in Giemsa and Gram stainings

501
Isolation and identification of D. congolensis
The bacteria grow on conventional media like sheep and ox blood
agar.
It growth is enhanced in 5 – 10 % Co2 atmospheric air.
So that candle jar is necessary.
Specimen for isolation of the bacteria is tuft of hair with scab
materials.
However, the scab materials contain many contaminates.
To over come this problem Haalstra’s method was developed.
Haalstra’s method for the primary isolation of Dermatophilus
congolensis
Grind up a small amount of scab material and place a little in 2ml
distilled water in a bijou bottle for 3.5 hours at room temperature.
Place the bijou bottle with lid removed , in a candle jar at room
temperature for 15 minutes.
 The motile zoospores are chemotactically attracted to the carbon
dioxide – enhanced atmosphere in the candle jar and move to the
surface of distilled water. 502
 Derm. cont’d---
 Then remove a loop full of fluid from the surface and inoculate on
blood agar plate.
Finally incubate the inoculated plate at 37 0C for 72 hours up to 5
days under 5 – 10 % Co2.
The colonies on blood agar after 24 – 48 hours are small 1mm in
diameter and have yellowish color.
They are haemolytic and are firmly embedded on the agar.
After 3 – 4 days the isolated colonies can be 3mm in diameter and
are rough, wrinkled and a golden - yellow color.
Glass Bijou (McCartney) bottles for microbiology

503
 Derm. cont’d---
Candle jars

504
Pure culture of D. congolensis on sheep blood agar after 3 days
incubation in 10% C o2

505
Differential diagnosis
Dermatophilosis must be differentiated from
Ring worm caused by fungi
Staphylococcal dermatitis/folliculitis
Scabies (mangemitis)
Pediculosis caused by lice
Fleece rot – sheep( mild superficial skin infection caused by the
combination of bacteria and moisture).
Treatment
Bactericidal dips are used to treat sheep from dermatophilosis.
However, it has limited efficacy as topical treatments do not
penetrate the scab to the active lesions.
The most commonly method that is used to treat sheep from
dermatophilosis is systemic administration of antibiotics.
Antibiotics that are effective include
 Procaine penicillin combined with streptomycin at a dose of 70 000
IU/ kg BW and 70 mg/kg BW respectively IM
506
Long acting tetracycline at dose of 20 mg/kg BW IM.
 Derm. cont’d---
Combination of lincomycin and spectinomycin at a dose of 5 mg/kg
BW and 10 mg/kgBW respectively IM.
Cattle treatment in temperate countries is done by administration
of : -
 Tetracycline at dose of 5 mg/kg BW IM and repeat weekly as
required is recommended
Long -acting tetracycline at the dose of 20 mg/kg BW IM in one
injection is reported to give excellent results in cattle.
Parenteral procaine penicillin at the dose of 22 000 IU/ kgBW
daily for three continuous days is also reported as efficacious
Dermatophilosis that occurs in tropical areas, the diseases is
associated with tick infestation.
Therefore, there is no completely satisfactory treatment in herds
with extensive involvement, or those being constantly re - infected
or exposed to predisposing causes.
In general terms, better results are obtained during dry weather and
in dry climates. 507
 Derm. cont’d---
So that parenteral treatment with antibiotics, as above can be used
and should be used in conjunction with acaricides when ticks are
present.
The most commonly used method to treat dermatophilosis in horses
is topical application of effective antiseptics.
To have effective treatment
 Horses should be free from whatever is causing prolonged wetness
of the skin.
Scabs must be removed by grooming under sedation
Then the lesions are treated topically daily with
Povidone-iodine or chlorhexidine until the lesions heal
 Benzoyl peroxide has keratolytic, antibacterial and follicular
flushing properties and is reported to be effective in therapy when
applied topically at a concentration of 2.5%
Severe cases can be treated daily for 3 days with penicillin at 20
000IU /kgBW IM alone or in combination with streptomycin at 10
mg/kg BW. 508
Prevention and control measures
The principal approach, where possible is, the avoidance of
predisposing factors.
The disease usually disappears in dry weather.
Isolation of infected animals and avoidance of contact by clean
animals with infected materials such as grooming tools is desirable.
Affected sheep should be shorn and/or dipped last.
Close yarding of sheep, or factors that promote face to skin contact
immediately after shearing or after dipping, should be avoided.
Insecticidal dips should contain a bactericide.
Because bactericidal dips will give some protection to sheep.
Some of bactericidal solutions that are commonly used are
 Spraying or dipping of sheep in 0.5 - 1.0% ZnSo4 (solution of zinc
sulfate) immediately after shearing is used to prevent infection of
shear cuts.
 Spraying or dipping sheep in a 1 % solution of alum (potassium
aluminum sulfate) provides protection against infection for up to
70 days as alum rendering the organism to be non-motile. 509
 Derm. cont’d---
An alternate treatment is to dust alum along the back of the sheep
If insecticidal dipping are factors for transmission dermatophilosis
infection and if there is no bactericidal agents, alternatively, pour-on
insecticides can be substituted.
In tropical areas, tick control is most important in control of
dermatophilosis.
Attempts for prophylaxis by vaccination in both sheep and cattle
have been unsuccessful.
Diseases associated with Moraxella species
Here we will see
Infectious bovine keratoconjuctivitis(IBK)
13. Infectious bovine keratoconjuctivitis(IBK)
Infectious bovine keratoconjuctivitis(IBK), sometimes referred to as
“ pink eye” or “blight”.
It is a highly contagious disease of cattle affecting the superficial
structures of the eyes, usually in animals under two years of age.
510
 Kerato cont’d---
The economic losses due to IBK disease arising from
Decreased weight gain
Loss of milk production
Treatment cost
The disease appears to be an aged – related immunity, probably as
the result of previous exposure.
Asymptomatic carrier animals harbour M. bovis in the naso -
lacrimal ducts, nasopharynx and vagina.
Etiology
Infectious bovine keratoconjuctivitis(IBK) is caused by Moraxella
bovis.
M. bovis is gram – negative rods or occasionally, cocci which
typically occur in pairs.
This organism is non – motile, aerobic and usually catalase and
oxidase positives.
The bacteria grow on conventional media like blood agar.
However, its growth is enhanced by the addition of sterile serum511
 Kerato cont’d---
into the medium.
The bacteria do not grow on MacConkey agar.
Virulent strains when isolated from cases of infectious bovine
keratoconjuctivitis are fimbriated and haemolytic.
Species other than M. bovis are periodically isolated from clinical
specimens are generally regarded as non – pathogenic.
Beta-haemolysin, pilin, leukotoxin and proteases are virulence
factors of M .bovis.
Betahemolysin is cytotoxic and produces corneal damage.
Studies on corneal tissue culture show a great variation in virulence
between strains.
M. bovis has serologically distinct shared and variable pilus
epitopes.
That is why strains of M. bovis can be distinguished by their pilus
antigens into seven distinct serogroups.
There are two distinct types of pilus.
They are I and Q (formerly α and β). 512
 Kerato cont’d---
 Q - pili mediate bacterial adhesion to the cornea and the
establishment of infection by preventing removal of the organism
by the continual flushing effect of ocular secretions and the
mechanical action of blinking.
Receptors for I-pili may be found on tissues other than the cornea
and facilitate colonization of M. bovis into non corneal tissues.
Epidemiology
The disease occurs in most countries of the world.
The young animals being the most susceptible animals.
But in a susceptible population, cattle of all ages are likely to be
affected.
There is no mortality due to IBK.
However, the morbidity rate can be as high as 80% and permanent
blindness or loss of an eye may occurs in some cases.
Severe outbreaks can be experienced in winter, especially if the
cattle are confined in close quarters such as barns or intensive
feedlots. 513
 Kerato cont’d---
Source of infection
Source of infection can be
Sick animals
Carrier animals
Cattle are the reservoir as the organism is carried on the conjunctiva
and also in the nares and vagina of cattle.
The bacteria can also colonize on the other tissues rather than
conjunctiva due to
 The presence of I pilli for attachment
 M. bovis can switch from expression of one pilus type to the other.
Transmission
The disease is most common in the end of winter and autumn
seasons and reaches epizootic proportions when flies and dust are
abundant.
Transmission is thought to be by means of these agents
contaminated by the ocular and nasal discharges of infected cattle.
The face fly (Musca autumnalis) and Asian face fly (Musca bezzii) 514
 Kerato cont’d---
because of their feeding preference around the eyes, are important
vectors.
M. bovis can be isolated from the crops of Musca autumnalis that
have fed on the eyes of infected cattle.
Risk factors of IBK
The most common risk factors for occurrence of IBK are
 1. Animal risk factors with their immune mechanism and
 2. Environmental factors
It is commonly observed that there is a much higher prevalence of
the disease in Bos taurus cattle as distinct from Bos indicus cattle.
Severity and proportion of bilateral infections is much greater in B.
taurus cattle than in crossbreeds and Bos indicus cattle.
Previous infection appears to confer a significant immunity that
lasts through to the next season.
Further reinfection usually results with minimal clinical disease,
confers the presence further immunity.
Because lacrimal secretions contain antibody, and antibody directed515
 Kerato cont’d---
against the pilus antigens of M. bovis will prevent adherence of the
organism to the cornea.
So that significant protection against IBK can be achieved by prior
vaccination with pilus antigens of the homologous strain.
However, there is antigenic diversity in pili from different strains of
M. bovis.
And vaccines composed of pili from one strain only confer
protection to challenge with organisms of the same serogroup.
But M. bovis in the eye can switch their pilus antigenicity in
response to antibody presence and render monovalent vaccines to be
ineffective.
So that a polyvalent vaccine might provide protection.
However, polyvalent vaccines are less immunogenic than
monovalent vaccines because of antigenic competition
The exposure of the eye to ultraviolet light, dust, to high
concentration of irritating gases(NH3, CH4 and H2S etc) may
increase susceptibility animals to the disease and the severity of 516
 Kerato cont’d---
Factors which may exacerbate or predispose to outbreaks of IBK
Factor Comments
Age Young calves less than two years of age are
particularly susceptible to IBK infection
Breed Bos taurus breeds appear to be more
susceptible than Bos indicus breeds
Fly activity Flies can act as vectors of M. bovis
Ocular irritants Dust with irritant gases, all grasses, grass
seeds, wind, UV light and cold ambient
temperature may predispose to disease
Concurrent infections Infection with bovine herpesvirus 1 or
Thelizia species may exacerbate IBK

Vitamin deficiency A deficiency of vitamin A may predispose to

IBK. 517
 Kerato cont’d---
signs resulting from it.
Pathogenesis
Attachment of M. bovis to the corneal epithelium is mediated by the
presence of pilus antigens.
It is known that Q-piliated organisms are more infectious than I –
piliated strains.
Microscopic corneal erosions are present within 12 hours of
infection and occur at this time in the absence of a significant
inflammatory response.
This indicates the initial production of the corneal ulceration is due
to the direct cytotoxic activity of the organism.
Then followed by focal loss of corneal epithelium, degeneration of
keratocytes, and invasion of the corneal stroma with fibrillar
destruction.
An inflammatory reaction occurs several days post infection and
518
 Kerato cont’d---
results in enlargement of the corneal ulcers with deeper stromal
involvement, corneal edema, and corneal neovascularization.
The lesions are localized in the eye and there is no systemic
infection.
Clinical findings
The usual incubation period of IBK is 2 - 3 days.
However, longer intervals up to 3 weeks have been observed after
experimental introduction of the bacteria to conjunctiva of cattle.
There are four stages of IBK
1. The first stage of IBK is characterized by
Edema of the conjunctiva accompanied by
A copious watery lacrimation
Blepharospasm
Photophobia
In some cases a slight to moderate fever.
Pain associated with pinkeye also decreases their feed intake.
As the result there is reduction in productivity. 519
 Kerato cont’d---
Stage I will progress to a small ulcer in the center of the cornea
which appears as a small white spot.
The cornea develops a slightly cloudy grey appearance due to
inflammation.
One or both eyes may be affected.
In 1 - 2 days a small opacity appears in the center of the cornea .
This may become elevated and ulcerated during the next 2 days.
Spontaneous recovery at this stage is quite common with and
without ulcer in the cornea.
About 2% of eyes have complete residual opacity.
The degree of ulceration in the early stages can be readily
determined by the infusion of a 2% fluorescein solution into the
conjunctival sac.
As the result the ulcerated area of cornea retaining the stain.
However, non ulcerated cornea doe not. 520
Cloudy cornea with conjuctivitis

521
Cloudy cornea with copious watery lacrimation

522
 Kerato cont’d---
NB. Most of IBK heal completely with a small white scar persisting
in the eye.
In the second stage of IBK the clinical signs described in stage I
continue.
That means with progressive of the disease the opacity becomes
quite extensive and the cornea becomes increasingly cloudy.
At the peak of the inflammation that is at about 6 days after the
appearing of the first signs , the inflammation may cover the entire
cornea
The color of the opacity varies from white to deep yellow.
However, at this point, some of the dark color of the iris can still be
seen.
Blood vessels from the outside portion of the cornea begin to grow
across the cornea to help in healing.
These blood vessels make the cornea to appear pink colour.
That is why IBK has another name called pink eye.
In the third stage of IBK;the ulcer covers most of the cornea and 523
Extensive cloudy of the cornea with vascularization

524
525
 Kerato cont’d---
the inflammation continues to spread into the inner parts of the eye.
When this occurs, the inner part of the eye fills with fibrin, which is
pus-like substance that gives the eye a yellow appearance versus
the typical brown appearance.

526
 Kerato cont’d---
In severe cases the cornea there becomes conical in shape, there is
marked vascularization of the cornea, and ulceration at the tip of the
swelling.
This is the four stage of IBK and characterized by
Extension of the ulcer completely through the cornea
 And the iris may protrude through the ulcer
As the result the iris will become stuck in the cornea even after
healing.
This may lead to glaucoma or persistent swelling of the eye.
This eye will be partially or completely blind.
The eye may go on to completely rupture, and will develop a
shrunken appearance or enlarge if glaucoma (increased eye
pressure) is present.
As the result this eye will be permanently blind.
527
Kerato cont’d---

528
 Kerato cont’d---
Young steer with IBK showing the healing stage with the
characteristic red cone of granulation tissue projecting from the
cornea

529
Kerato cont’d---

530
 Kerato cont’d---
Lastly, once healing occurs (except stage IV) the blood vessels will
recede.
But the eye may continue to be a cloudy blue color.
The blue appearance may eventually resolve and the eye appears
clear again.
In other cases, depending on the severity of the disease, a white scar
may be present even after full resolution of the disease.

531
Necropsy findings
Necropsy examination is not usually necessary.
Diagnosis
Diagnosis of IBK is done based on
Epidemiology
Clinical signs
Laboratory
The organism can be isolated by
Culturing on blood agar
Lacrimal secretion is the most suitable specimen for laboratory
examination.
For transportation of swabs of lacrimal secretion should be placed
in 1 to 2 ml of sterile water.
Ideally, specimens should be cultured within 2 hours of collection.
On blood agar, the colonies are round, small, shiny and colonies of
virulent strains are surrounded by a zone of complete haemolysis.
Smears from colonies reveal short gram negative rods in pairs.
No growth on MacConkey agar. 532
 Kerato cont’d---
Reaction in catalase and oxidase tests are positive.
A loeffler’s serum slope may be pitted after 10 days.
Gram negative rods of M. bovis in pair Loeffler’s serum slant

533
 Kerato cont’d---
Additionally the bacteria can be identified from lacrimal specimens
by fluorescent antibody technique(FAT).
Serologically, serum agglutinins (1:80 to 1:640) are present 2 - 3
weeks after clinical signs commence.
And gar gel immune diffusion test is capable of detecting M. bovis
antibodies after 2 – 3 weeks of infection.
Differential diagnosis
IBK must be differentiated from
 1. Traumatic conjunctivitis
Traumatic conjunctivitis is easily differentiated because of the
presence of foreign matter in the eye or evidence of a physical
injury.
 2. Pasteurella multocida infection(capsular type A)
It can be differentiated by laboratory diagnose.
3. Severe conjunctivitis with corneal opacity and ulceration caused
Mycoplasma bovis.
It is differentiated by serological conversion in affected animals. 534
 Kerato cont’d---
In mycoplasmal keratoconjuctivitis the involvement of the eyelids
with marked swelling is prominent than IBK.
That means conjunctivitis is prominent in mycoplasmal infections
that produce keratoconjuctivitis.
4. Listeria monocytogenes iritis
5. Infectious bovine rhinotrachitis
6. Bovine malignant catarrh infection
7. Thelaziasis
8. Chlamydial keratoconjuctivitis
Chlamydial keratoconjuctivitis presents with identical clinical
findings with IBK.
But has a protracted course despite treatment and a higher morbidity.
Chlamydophila DNA can be detected by conjunctival swabs.
This disease is a possible zoonosis. 535
Treatment
Early treatment of cattle with pinkeye is important because
It helps not only for a successful outcome of treatment of the
affected animals.
But also to stop the shedding of the bacteria to decrease the risk of
transmission to other cattle.
The principle of treatment depends on the stage of IBK
In the first stage it is advisable to use
 Long-acting tetracyclines and the recommended dose is 4.5 ml/ 45
kg BW IM.
 A second injection given 48 to 72 hours later may increase the
percentage of cattle that responds to treatment.
 Another option is to inject penstrip and anti-inflammatory drug
like dexametazone into the bulbar conjunctiva.
The bulbar conjunctiva is the thin membrane that covers the white
portion (or sclera) of the eye.
If the injection is performed correctly, the conjunctiva will swell
and a bulge should be seen in this area. 536
 Kerato cont’d---
Bulbar conjunctival injections

537
 Kerato cont’d---
In this stage the bacteria is sensitive for topical application of
ophthalmic ointments that' containing antimicrobials like
antibiotics and sulfonamides.
But they need to be instilled in the conjunctival sacs at frequent
intervals, which may be impractical under field conditions.
In the second stage of IBK it is advisable to use
 Both tetracycline and a bulbar conjunctival injection are
administered at the above dosages.
In the third stage of IBK also
Tetracycline and a bulbar conjunctival injection are administered in
conjunction with either an eye patch or suturing the eyelids shut.
This makes the eye more comfortable, reducing further irritation
and therefore, reducing tearing and shedding of the bacteria.
In the fourth stage of IBK the same treatment as the third stage
must be performed
In addition of topical application it is good to use parenteral therapy
538
An eye patch

539
 Kerato cont’d---
with
Sulphadimidine at the normal dose rate of 100 mg/kg BW. or
A single treatment with long-acting oxytetracycline (20 mg/kg
intramuscularly) has a good efficacy.
Prevention and control measures
Fimbrie – derived bacterins are available commercially in some
countries but have uncertain efficacy.
Effective management methods are important to control IBK.
These includes
On time isolation of affected animals
Reduction of exposure to mechanical irritants( dusts, irritant gasses)
Use insecticidal ear tags
Control of concurrent diseases such as infectious bovine rhino –
tracheitis or thelaziasis.
Vitamin A supplementation may be beneficial.

540
Diseases associated with Mycoplasma spp.
Under this genus we will see
 Contagious Bovine Plueropneumonia(CBPP)
Contagious Caprine Plueropneumonia (CCPP)
13. Contagious Bovine Plueropneumonia(CBPP)
CBPP is a sever contagious disease of cattle characterized by
localization of infection in the lungs and pleura characterized by
pneumonia and pleuritis( pleurisy).
Etiology
CBPP is caused by Mycoplasma mycoides subspecies mycoides.
Mycoplasma mycoides subspecies mycoides is found under genus
Mycoplasma.
The positive agent also called Mycoplasma mycoides subsp.
mycoides small colony (MmmSC).
The organism belongs to the 'mycoides' cluster which consists of six
closely related species Mycoplasma.
Members of the 'mycoides' cluster are pathogens of cattle, sheep
and goats. 541
 CBPP cont’d---
Members of Mycoplasma mycoides cluster
Name Main disease Main( and other) hosts
M. mycoides subsp. mycoides SC Contagious bovine Cattle (goats, sheep,
variant plueropneumonia buffalo)

M. mycoides subsp. mycoides LC Caprine pneumonia, Goats (sheep, cattle)

variant contagious agalactiae

M. mycoides subsp. capri Caprine pneumonia Goats but rare in sheep

M. capricolum subsp. capricolum Caprine pneumonia, Goats (sheep, cattle)

contagious agalactiae
M. capricolum subsp. Contagious caprine Goats (sheep)
capripneumoniae plueropneumonia

M. bovine group 7 (Bg7) Arthritis, mastitis and Cattle

calf pneumonia

542
 CBPP cont’d ---
Among the six clusters only two of them cause disease in cattle.
These are
 Mycoplasma mycoides sub spp. mycoides of small
colony(MmmSC )which is the cause of CBPP and
 Mycoplasma bovine group 7 (Bg7) which causes arthritis and
bovine mastitis.
The agent of contagious bovine plueropneumonia (CBPP) also
called small colony types is not communicable to other species.
Large colony types are pathogenic for sheep and goats but not for
cattle.
However, the small and large colony types can be differentiated
culturally and biochemically.
The organisms are pleomorphic and some forms are filterable.
They can be maintained readily in
Special culture medium called beef( heart) infusion medium with
the addition of pooled serum or
 In embryonated hens' eggs 543
 CBPP cont’d ---
In beef( heart) infusion agar they form microscopic( small colony)
that have fried egg appearance.
Colonies of mycoplasmas on agar plates which has fried egg
structure

544
Epidemiology
Under natural conditions, CBPP affects
All species of cattle (Bos indicus and Bos taurus) of all age groups
Allied animals including buffalo, yak, bison and even reindeer.
CBPP is eradicated in most countries of western hemisphere
including Australia.
But it is endemic in Africa and less occurs in some countries of Asia
and Europe.
There was a very high incidence of CBPP in Zambia, Tanzania,
Botswana
Newly infected areas in the 1990s include Uganda, parts of Kenya,
Region of the Democratic Republic of the Congo, Tanzania, where
recently the disease was eradicated.
Lesotho, Malawi, Mozambique, South Africa, Swaziland, and
Zimbabwe are currently free (2002 OIE report)
But countries which were free from CBPP are reinfected again by
the disease.
Totally the incidence of CBPP in most African countries has 545
 Myco cont’d ---
increased alarmingly this is because of
Lack of a clear disease-control policy
Uncontrolled cattle movements
 Lack of public awareness
Lack of commitment and ineffective legislation
CBPP is less prevalent in European and Asian countries like Spain,
Portugal, Italy, Middle East, India, and China, Bangladesh and
Myanmar.
Outbreaks of CBPP tend to be more frequent in intensive farms
and in those in transit by train or on foot.
Morbidity because of CBPP increases with close confinement and
reaches 100% in susceptible herds.
The case mortality may be as high as 50%.
From of the rest of 50%, 25 % of the infected cattle remain as
recovered carriers with or without clinical signs.
Source of infection
Sources of infection for CBPP are 546
 CBPP cont’d ---
Sick animals
Recovered 'carrier' animals.
Recovered animals can be source of infection as pulmonary
sequestrum preserves a potential source of organisms for periods as
long as 3 years.
For many years, it was thought that conditions of stress due to
starvation, exhaustion or intercurrent disease can cause the
sequestrum to break down and convert the animal into an active
case.
But experimental evidence throws some doubt on this explanation.
Eventhough droplet infection is usually associated with a donor
lesion in the lungs.
Renal lesions are not uncommon and large numbers of viable M.
mycoides are passed in the urine of infected animals, and inhalation
of urine droplets may be a route of infection.
Methods of transmission
The organism is present in saliva, urine, fetal membranes, uterine
547
 Myco cont’d ---
discharges and fetal membrane of affected animals.
Transmission occurs from direct and repeated contacts between
sick( carrier) and healthy animals.
The principal route of infection is by inhalation of infective
droplets from active or carrier cases of the disease.
Mediate infection by contamination of inanimate objects is unlikely
under natural conditions.
However, it has been effected experimentally, the infected hay
remaining infective for up to 144 h.
Other inanimate objects such as placenta and urine can also remain
infective for long periods.
It has been suggested that urine may be a mode of transmission
especially in intensive farms.
Where cattle are reared intensively in restricted areas.
Transplacental infection is also reported.
Human being is not susceptible to CBPP infection( not zoonotic).
548
 CBPP cont’d ---
Risk factors
The risk factors for occurrence of CBPP are
1. Animal risk factor
2. Management risk factor
 3. Pathogen risk factor
1. Animal risk factor
CBPP affects only cattle; rare natural cases have been observed in
buffalo, yak, bison, reindeer and antelopes.
Wild bovids and camels are resistant.
The disease has been produced experimentally in captive African
buffalo and white tailed deer.
It has not been detected in other wildlife.
There is no difference in the susceptibility of Bos taurus and Bos
indicus cattle.
A strong immunity develops after an attack of the natural disease in
cattle.
So that vaccination plays an important part in control of CBPP. 549
2. Management risk factor
The occurrence and incidence of CBPP is heavily influenced by
Management systems(intensive, semi – intensive and extensive)
Disease control policies and regulations of a country
 Knowledge of the disease by farmers and veterinarians and
livestock field officers
Diagnostic capability of veterinary laboratories
Disease-surveillance and monitoring systems
Adequacy of vaccination programs
Government budgets allocated to control the disease
3. Pathogen risk factor
M. mycoides subsp. mycoides is sensitive to all environmental
influences, including disinfectants, heat and drying.
That is why it does not ordinarily survive outside the animal body
for more than a few hours.
A low incidence can be anticipated in arid regions because of the
rapid destruction of the organism in exhaled droplets.
The lack of a cell wall and endotoxins may enable mycoplasmas 550
 CBPP cont’d ---
to colonize the animal without inducing an immune response.
The predilection site of the organism is the mucosal membranes
that limits the humoral response of the affected cattle.
For these reasons it is suggested that the organism is a poor
immunogenic which may account for the frequent lack of good
circulating antibody.
A separation of 6 m between animals is usually considered to be
sufficient.
However, transmission over 45 m has been suspected to occur.
Economic importance of CBPP
CBPP is the most economically important disease of cattle in
Africa.
Because of direct and indirect economic losses.
The direct losses are from
Mortality
Reduced milk yield
Vaccination costs to control the disease 551
 CBPP cont’d ---
Cost for disease surveillance and research programs
The indirect losses are due to the chronic nature of the disease
including:-
Loss of weight and working ability
 Delayed marketing
 Reduced fertility
 Losses due to quarantine
Loss due to restriction of international cattle trade
In the affected countries, enormous losses are experienced each
year from
 Deaths of animals
 Loss of production during convalescence
The highly fatal nature of the disease, the ease of spread and the
difficulty in detecting carriers makes to have a close restriction on
the movement of animals from enzootic areas.
Pathogenesis
552
CBPP causes an acute lobar pneumonia and pleurisy.
 CBPP cont’d ---
The organism invades the lungs of cattle and causes a
mycoplasmemia.
This results in localization in numerous other sites including the
kidneys and brain.
This results in high morbidity and mortality of infected animals.
Thrombosis of pulmonary vessels is an essential part in
pathogenesis of CBPP.
It is formed probably prior to the development of pneumonic lesions.
The mechanism of development of the thrombosis in pulmonary
vessels during CBPP is not well understood.
However, there is no general increase in blood coagulability, and no
generalized tendency to spontaneous thrombosis.
The production of hydrogen peroxide and other active super oxides
is widely believed to play an important role in mycoplasmas
pathogenicity.
These peroxides and super oxides cause lysis of erythrocytes,
peroxidation of lipids and inhibition of ciliary movement in trachea.
553
 CBPP cont’d ---
Death results from anoxia and presumably from toxemia.
Clinical findings
There is considerable variation in the severity of CBPP.
So that CBPP can have
1. Peracute
2. Acute
3. Subacute and chronic forms
1. Peracute form of CBPP
In peracute form of CBPP the affected cattle may die within 1 week
after the onset of respiratory distress.
2. Acute form of CBPP
Incubation period lasts 3 - 6 weeks (occasionally up to 6 months).
There is
A sudden onset of fever upto 400C
 Sudden reduction of milk yield
Anorexia, depression and cessation of rumination
Animals stand apart or lag behind a traveling group 554
 CBPP cont’d ---
Coughing at first only on exercise
 Thoracic pain
Affected animals are disinclined to move, standing with the elbows
out, the back arched and head extended.
Respirations are shallow, rapid and accompanied by expiratory
grunting.
Pain is evidenced on percussion of the chest.
 Auscultation reveals pleuritic friction sounds in the early stages of
acute inflammation, and dullness fluid sounds and moist gurgling
crackles in the later stages of effusion.
Dullness of areas of the lung may be detectable on percussion.
Edematous swellings of the throat and dewlap may occur
 Swelling of the large movable joints may be present
In calves, valvular endocarditis and myocarditis may occur.
In fatal cases death occurs after several days upto 3 weeks.
3. Subacute and chronic form of CBPP
Recovered animals may be clinically normal. 555
 CBPP cont’d ---
But in some animals there may be inactive sequestrum forms in the
lung.
The sequestrum with a necrotic center of sufficient size to produce a
toxemia which can result in un thriftiness, a chronic cough and mild
respiratory distress on exercise.
These sequesters commonly breakdown when the animal is
exposed to environmental stress and cause an acute attack of the
disease.
Necropsy findings
Lesions are confined to the thoracic cavity and lungs.
Lesions on lungs are usually unilateral.
The pleural cavity may contain large quantities of clear, yellow-
brown fluid containing pieces of fibrin .
Caseous fibrinous deposits are present on the parietal and visceral
surfaces of the lungs.
The interlobular septae of lung are prominently distended with
amber-colored fluid surrounding distended lymphatics. 556
 CBPP cont’d ---
This fluid distinctly outlines the lobules with vary in color with
red, gray, or yellow hepatization.
One or both lungs completely or partially affected with marked
consolidation.
Affected lobules show various stages of grey and white
hepatization.
The interlobular septa greatly distended with sero fibrinous
exudates giving rise to the classical marbled lung which is the
typical necropsy findings during CBPP.
Adhesions between the pleural surfaces are constant findings of
CBPP.
In chronic or advanced cases a sequestrum of necrotic lung
varying size from 1 - 10 cm in diameter is surrounded by a fibrous
capsule are present.
So that, if these sequestrae rupture and are drained by a bronchus
they can be a source of aerosol infection to cattle.
Such a mechanism accounts for epidemics in closed herds. 557
 CBPP cont’d ---
In affected calves, exudative peritonitis, arthritis, bursitis and
fibrinous arthritis of carpal and tarsal joints may be present.
The joint may be enlarged due to a proliferation of connective tissue
and tendosynovitis (inflammation of the tendon and its sheath).
Renal lesions are frequently detectable in CBPP.
In the acute phase of the disease there is multiple renal infarcts.
 In subacute and chronic cases the infarcts progress to form large
areas of fibrosis accompanied by tubular dystrophic calcification,
tubular atrophy with lymphocyte interstitial infiltrates.
Diagnosis
There are many causes of severe pneumonia in cattle
So that it is difficult to diagnose CBPP based on clinical signs alone.
It is better to diagnose CBPP by considering the epidemiology,
clinical signs, necropsy findings and laboratory diagnosis.
Animals with CBPP frequently present with unilateral pneumonia
and in a herd with signs of pneumonia in adults and polyarthritis in
calves. 558
 CBPP cont’d ---
Post mortem lesions are often helpful in diagnosis of CBPP, like
 Presence of sero - fibrinous exudates with classical marbled lung
 Presence of sequestrum of necrotic lung varying size from 1 - 10
cm in diameter is surrounded by a fibrous capsule in chronic cases.
Confirmatory diagnosis is done by isolation and identification of the
agent.
The specimens of choice are
Pleural effusion fluid in serum tube
 Affected Lung
Bronchial lymph node
The organism is nutritionally very fastidious and special laboratory
media is required for growth and identification
Final identification of mycoplasmas is usually made by
Growth inhibition or
Immunofluorescence tests on agar
 Latex agglutination test(latex microspheres coated with anti-
559
MmmSC IgG)
 CBPP cont’d ---
Polymerase chain reaction (PCR) has been used to identify the
specific organism and differentiate it from other members of the
cluster
Serology
The most common serological tests that are used to diagnose CBPP
are CFT and ELISA
 CFT is used to detect early infection
ELISA is more sensitive to detect late infection.
Differential diagnosis
CBPP must be differentiated from
1. Rinderpest
Rinder pest is differentiated from CBPP in that there is erosive
stomatitis, dysentery and erosions throughout the alimentary tract
but not in CBPP.
2. Foot and mouth disease(FMD)
In FMD, there is salivation, lameness, fever, and vesicular
stomatitis but not in CBPP. 560
 CBPP cont’d ---
3. Hemorrhagic septicemia in cattle
Hemorrhagic septicemia is an acute disease with death in 6 to 72
hours.
It spreads more rapidly through a herd than CBPP.
Lung lesions during hemorrhagic septicemia are almost similar to
the lung lesions that are see in the early stage of CBPP.
But there is no classical marbled lung and sequestrum of necrotic
lung in hemorrhagic septicemia that are present in later stage of
CBPP.
Lastly we can differentiate them in laboratory by culturing.
4. Theileriosis (East Coast Fever)
In east cost fever there is coughing, nasal and ocular discharge,
diarrhea, enlargement of peripheral lymph nodes, ulceration of
abomasum which are not present in CBPP.
Absent lung lesions in theileriosis.
5. Pulmonary abscesses
561
 CBPP cont’d ---
Large abscesses containing foul-smelling purulent material caused
by different bacteria may have total destruction of lung but not
CBPP.
6. Tuberculosis
Tubercular nodules may resemble CBPP sequestra.
However, tubercular nodules are degenerative cheese-like lesions,
often calcified.
7. Actinobacillosis
Generalized lesions of lung and other adjacent tissues but not in
CBPP.
8. Echinococcal (hydatid cysts)
Pulmonary cysts with a double wall and containing
clear fluid, often calcified when old.
Treatment
Treatment of CBPP is recommended only in endemic areas.
Because elimination of the organism may not be possible and
carriers may develop. 562
 CBPP. cont’d ---
Antibiotic treatment is generally not effective as it can result in
extensive tissue damage and sequestration of the organism.
As soon as an outbreak is suspected, slaughter and necropsy of a
suspect animal is advisable.
An “in vitro” trial of five commonly used antibiotics in treatment of
CBPP on recent isolates of MmmSC found that
Tilmicosin and danofloxacin were effective both in terms of
mycoplasmastatic and mycoplasmacidal activity.
Tylosin, florofenicol and tetracycline were intermediate
Spectinomycin was ineffective against some strains MmmSC.
Dose and route of administration
Tylosin (10mg/kgBW every 12hrs for six injection)
Spiramycin (10 - 50mg/kgBW for 3 days ) are effective in the
treatment of clinical cases
Prevention and control measures
The major obstacles to control and to eradicate CBPP in sub –
563
Sahara countries are:
 CBPP cont’d ---
Difficulty in controlling the movements of cattle
Complications of applying quarantine and slaughter policies
 Lack of rapid pen-side diagnostic tests
 Ineffective vaccines
 Insufficient funds to implement control policies
 Civil strife and drought, which have an effect on the spread of the
disease in Africa.
The possible strategies that are used to control CBPP in CBPP
endemic countries or regions are strategies on the country and
region basis and herd level.
On the country level
1. Slaughter of all sick and in-contact cattle.
This strategy is impractical in developing countries with a pastoral
economy.
2. Slaughter of all sick cattle and vaccination of in-contact cattle
This strategy is used frequently and usually perpetuates the disease
564
in many countries.
 CBPP cont’d ---
 3. Vaccination of healthy cattle with slaughter of sick cattle in
an epidemic and revaccination of cattle at risk.
This method depends on the ability of the authorities to detect
epidemics rapidly most effectively by abattoir surveillance and to
maintain vaccination for at least 3 years.
Vaccination in endemic areas must be done annually
While newly infected areas require repeat vaccinations aimed at
eradication of the disease after 3, 9, 21, and 36 months.
Specific therapy is prohibited.
On an area basis
The prevention of entry of infected animals into a free area is a
difficult task.
Only the following classes of cattle should be permitted to enter:
1. Cattle which have not been in an infected area nor in contact
with infected animals for at least 6 months should be permitted
for entery.
However, cattle going to immediate slaughter after a clinical 565
 CBPP cont’d ---
examination if CBPP is suspected within a period of 1 month.
2. Cattle which have given negative reactions to the CFT on two
occasions within the preceding 2 months and have not been in
contact with infected animals during this period.
These animals may or may not have been vaccinated.
It is less rigid measures because these will permit introduction of
the disease.
When the disease is already present in an area, two methods of
control are possible.
 These are vaccination and eradication by test and slaughter of
reactors.
The method chosen will depend largely on the economy
of the cattle industry in the affected area.
On herd basis
When the disease becomes established in a herd the following
measures may be adopted to prevent its spread.
1. Removal of sources of infection 566
 CBPP cont’d ---
2. Hygiene
3. Vaccination
1. Removal of sources of infection
Infected animals should be removed from the herd as soon as
possible.
The CFT is adequate to identify the infected animals and should be
carried out in conjunction with clinical examination.
2. Hygiene
1. Any procedure which brings the animals together should be
avoided especially in humid conditions when droplet inhalation is
more likely to occur like
Passage through the milking shed
Collecting of animals for inspection and marketing
Bleeding and vaccination
 2. Strict quarantine of the infected and in-contact herds must be
maintained until all residual infection has been eliminated.
It is usually 12 weeks after the removal of the last reactor and/or567
 CBPP cont’d ---
clinical case is sufficient time.
 Animals in quarantine should be kept under constant surveillance so
that clinical cases may be observed.
3. Vaccination
All effective CBPP vaccines are live attenuated vaccines.
They are prepared from freeze dried broth cultures of Mycoplasma
mycoides subsp. mycoides SC type(MmmSC)
Current vaccine strains that are used now are
T144
T1 SR
Generally these vaccine considered to exhibit poor efficacy and
stability because
Protection is low (only 30 to 60% of animals are protected)
Immunity lasts duration (6 - 12 months) so that repeated
vaccination is necessary.
A number of post vaccine reactions may occur for example within
two to four weeks following vaccination an invading edema 568
 CBPP. cont’d ---
develops.
 The reversion to virulence specially T1 44 vaccine has also been
observed.
 As the result vaccinated animals by T1 44 and T1 SR
subcutaneously can be reservoirs for MmmSC and infect other
animals in areas where previously free of CBPP.
The value of calf hood vaccination is limited.
Because arthritis, myocarditis and valvular endocarditis occur 3-4
weeks after vaccination of calves less than 2 months old.
So that vaccination of calves must be started after two months of
age.
The available vaccine to control CBPP in Ethiopia is
freeze – dried live attenuated bacterial vaccine produced using T144
and T2 SR strains of Mycoplasma mycoides subspp. mycoides
small colony variant(MmmSC) .
Which is available in 100 doses per vial.
Reconstitute the freeze – dried vaccine by sterile saline. 569
 CBPP cont’d---
Dose – 1ml S/C/ and vaccinate all animals above 6 months of age.
Immunity lasts almost one year.
14. Contagious caprine plueropneumonia (CCPP)
It is a contagious mycoplasmal disease of goat characterized by
Pneumonia
 Fibrinous pleurisy
 Profuse pleural exudate and marbled appearance on the cut surface of
affected lungs.
Although similar in many respects to CBPP, well developed necrotic
areas in the lungs in chronic CCPP are rare.
CCPP is one of the most serious fatal diseases of goats.
It is commonly confused with other serious pneumonias of goats and
sheep.
However, it presents primarily as a plueropneumonia.
Etiology
Contagious caprine plueropneumonia (CCPP) is caused by
Mycoplasma capricolum subsp. capripneumoniae.
570
 CCPP. cont’d ---
The causative agent is previously known as mycoplasma strain F
38.
The organism does not survive long outside the host body.
This organism is difficult to grow which has led to poor
differentiation of the disease from that induced by M. mycoides
subsp. mycoides LC and M. mycoides subsp. capri.
There are different strains and one study established four different
lineages of M. capricolum subsp.capripneumoniae based on
nucleotide sequence.
Epidemiology
The exact distribution of the disease is unknown.
However, Reports of CCPP come most commonly from northern
and eastern Africa, Spain, Turkey, Mediterranean littoral, from
many countries of Asia and India.
But clinical disease has been reported from 38 countries, mostly
from Africa and Asia.
571
CCPP has many similarities clinically and at necropsy to
 CCPP cont’d ---
But it is not transmissible to cattle.
Infectivity is high with a morbidity of 100% and the illness is
acute and severe with a case mortality rate of 60-100 % .
Morbidity and mortality rates in a recent outbreak in a herd in
Eretria were 90 and 65% respectively.
Source of infection
Source of infection in CCPP can be
Infected sick animals
Carrier animals( latent infection)
Transmission
The disease is readily transmitted by inhalation( aerosol).
However, the organism does not survive for long outside the animal
body.
So that direct contact of infected animals( carrier animals) with
susceptible one is necessary.
Clinical findings
The clinical findings in contagious caprine plueropneumonia are 572
 CCPP. cont’d ---
restricted to the respiratory system.
The prominent clinical findings are
Fever (40.5 – 410C)
Cough
Dyspnea
 Lagging
 Lying down a lot (but the animal can stand and walk), fever
In terminal stage mouth-breathing, tongue protrusion and frothy
salivation with death in two or more days.
Under adverse climatic conditions the disease may occur in a
septicemic form with little clinical or postmortem evidence of
pneumonia.
NB. M. mycoides subsp. mycoides LC and M. mycoides subspp.
capri may cause pneumonia with similar clinical manifestations
like CCPP. However, in necropsy finding there is only
pathological change in lung tissue but not in pleura.
573
 CCPP. cont’d ---
Necropsy findings
The more usual necropsy findings are similar to those of contagious
bovine plueropneumonia.
However, sequestra are not formed in the lungs during CCPP.
Lesions are restricted to the lungs and pleura with hepatization of parts of
the lung and an increase in pleural fluid with a fibrinous pleuritis.
Histologically, contagious caprine plueropneumonia is characterized by an
interstitial interlobular edema rather than ‘ a thickening of the interlobular
septa seen with other mycoplasmal infections.
The lesions may be confined to one lung.
Diagnosis
Diagnose of CCPP can be done by considering
Epidemiology of the disease
Clinical findings
Necropsy findings
 Bacteriological findings
Molecular techniques 574
 CCPP. cont’d ---
 Serology
Laboratory diagnose is important to isolate and identify the positive
agent.
Molecular techniques like PCR is used for detection of antigen in
lung tissues and pleural fluids.
The serological tests that are used to identify carrier animals
include
Complement Fixation Test(CFT)
ELISA
Latex agglutination test
Latex agglutination test is available commercially and suitable for
field use.
The F38 monoclonal antibody is used in serological tests to identify
caprine F38-type isolates by the disc growth inhibition method
A blocking ELISA using monoclonal antibodies is highly specific
for CCPP
575
 CCPP. cont’d ---
Differential diagnosis
CCPP must be differentiated from other pulmonary mycoplasmosis
that are caused by
M. mycoides subsp. mycoides SC variant causes CBPP
M. mycoides subsp. mycoides LC variant causes caprine pneumonia
and contagious agalactiae
M. mycoides subsp. capri causes caprine pneumonia
M. capricolum subsp. capricolum causes caprine pneumonia and
contagious agalactiae.
Treatment
CCPP can be treated by using
 Tylosin - 17.5 mg/kg BW IM for 3 - 5 consequentive days or
Oxytetracycline - 15 mg/kg BW IM for 3 consequentive days
They are highly successful in limiting the severity of disease.
However, eventhough the severity of the disease is reduced, treated
animals are still sources of infection.
576
 CCPP. cont’d ---
Prevention and control measures
Herd biosecurity to prevent contact with infected animals is
important.
Vaccination with an inactivated mycoplasma F38 vaccine induces an
immune response which is effective in reducing morbidity and
mortality rates.
Booster dose 1 month after the first vaccination provides additional
protection.
Immunity is generally short-lived.
A vaccine for CCPP is present in Ethiopia.
It is inactivated saponin adjuvanated bacterial vaccine produced
using F -38 Kenyan strain of Mycoplasma capricolum subspp.
capripneumoniae.
The vaccine is a liquid suspension and available in 100 ml for 100
doses.
Dose and route of administration
Dose 1ml/ goat S/C in the inner thigh of goats. 577
 CCPP. cont’d ---
Vaccinate goats above 6 months of age.
Immunity lasts for 1 year.
NB. New born kids must not be vaccinated before 6 months of age
because maternal antibody may interfere with vaccine.
15. Listeriosis
The name Listeriosis is derived from a genus of bacteria Listeria that
was named after the English pioneer of sterile surgery Joseph Lister in
1940.
Listeriosis is bacterial infectious disease of animals characterized
by
Encephalitis
Abortion
Septicemia
Endophtalmitis and mastitis
Usually only one form of the disease occurs in a group of affected
animals.
Septicemic form of listeriosis often encountered in a new born 578
 List. cont’d---
piglets, foals, cage birds and poultry can also occur in adult sheep.
Listeriosis is a zoonotic disease.
Etiology
The genus Listeria has been divided into seven species with two
distinct groups.
The first group comprises
L. murrayi
L. grayi
 L. seeligeri
 L. innocua
 L. welshimeri
All these species of Listeria are non – pathogenic groups.
NB. Even if L. innocua is grouped under non – pathogenic group of
genus Listeria it may cause rarely meningoencephalitis in sheep.
The second groups comprises
 L. monocytogens
 L. ivanovii 579
 List. cont’d---
L. monocytogens are pathogenic for animals and human beings.
Listeria ivanovii is a pathogen of mammals specifically ruminants
and rarely it causes listeriosis in human being.
L. monocytogens is widespread in nature and has characteristics that
allow its survival and growth in a wide variety of environments.
Optimal growth temperatures are between 30°C and 37°C.
But the organism can grow and reproduce at temperatures between
-0.4°C and 45°C.
It can grow at PH between 4.5 and 9.6 and at low temperature
although growth at low PH and temperature is minimal.
The organism is susceptible to common disinfectants.
L. monocytogens can be divided into 16 serovars on the basis of
somatic(O) and flagellar (H)antigens.
Serovars 4b, 1/2a and 1/2b and 3 are most commonly isolated from
diseased animals
Virulent strains can multiply in macrophages and monocytes and
580
monocytes.
 List. cont’d---
Haemolysin, listerolysin 0 are believed to be a major virulence factor for
pathogenic species of Listeria.
Means of transmission of listeriosis to human beings as it is zoonosis

Epidemiology
Although the organism is widespread in nature, clinical disease in animals
occur seldom in tropics and sub tropics.
Because most but not all outbreaks of listeriosis in animals is associated
with silage feeding of animals.
581
 List. cont’d---
But listeriosis is important in temperate zones of the world mostly
in North America, Europe, New Zealand and Australia.
In the northern hemispheres listeriosis has a distinct seasonal
occurrence.
As it is associated with seasonal feeding of silage with the highest
prevalence in the months of December through May.
Listeriosis in animal
Listeriosis is primarily a disease of ruminants particularly sheep
and cattle and rarely in other species of animals.
The major diseases those are associated with L. monocytogenes and
rarely with L. ivanovii are
1. Visceral (septicemic) listeriosis in many species of young animals
2. Neural listeriosis that can be because of
Encephalitis called circling disease with micoabscess in the
brainstem
Spinal myelitis
3. Abortion 582
 List. cont’d---
4. Iritis
 5. Rarely gastroenteritis and mastitis
The main diseases of animals caused by genus Listeria
Species of Listeria Affected species of animals Disease syndrome

Young animals of many Visceral (septicemic)

species listeriosis

Neural listeriosis: circling

L. monocytogens disease with micoabscess in
Sheep, goat and cattle the brain stem.
Spinal myelitis
Abortion

Cattle Iritis

L. ivanovii Sheep and cattle Abortion

583
Source of infection
The organism is common in the environment.
The ability to form biofilms(slime layers) may assist in
its survival in the environment.
Biofilms may assist in perperuating its presence in water troughs on
infected farms.
Most feed hays, grains and formulated feeds have the potential to
contain L. monocytogenes.
In ruminants L. monocytogenes can be isolated from the feces and
nasal secretions of healthy animals.
The organism may be present in bulk tank milk or milk filters.
The presence of L. monocytogenes in bulk tank milk or milk filters
is used as a measure of farm infection prevalence.
The main source of infection in domestic animals are
 1. Poorly fermented silage with the PH of 5.0 - 5.5( source of the
organism is contaminated soil)
 2. Big bale silage(source of the organism is contaminated soil as it
has greater risk for mechanical damage to the plastic covering. 584
 List. cont’d---
 3. Moist preserved feeds other than grass silage are at risk for
listeria growth. For example
Moist brewers' grains,
 Wet spoiled hay bales and
Silage made from commodity by-products such as orange and
artichoke waste.
4. Infective materials( inanimate objects) that are contaminated by
feces, urine, aborted fetuses and uterine discharges and milk
Transmission
The main means of transmission are
 Ingestion of contaminated material, milk , poorly fermented silage,
big bale silage and moist preserved feeds.
 Through the navel from the environment and
 Congenital infection of the fetus
Risk factors
Despite the ubiquity of L. monocytogenes only a small proportion of
animals develop clinical disease. 585
 List. cont’d---
A number of predisposing factors have been proposed as risk
factors for disease.
These include
 1. Factors that cause a lowering of the host animal's resistance like
host management risk factor and
 2. Factors that increase the infection pressure of the organism
called pathogen risk factor
In farm animals the latter appear the most important.
1. Host and management risk factor
Concerning the host, all breeds of animals are susceptible to
listeriosis.
However, a few studies show angora goats and rambouillet sheep
are more susceptible than other species of goats and sheep.
There are many risk factors which are associated with management
and can predispose animals to listeriosis,
Some of them are
 Poor nutritional state 586
 List. cont’d---
Sudden changes of weather to very cold and wet
The stress of late pregnancy and parturition
Transport
 Long periods of flooding with resulting poor access to pasture.
Listeriosis outbreak affecting several flocks can occur in sheep on
poorly drained and muddy pastures following floods.
However, outbreaks are also described in droughts.
Overcrowding and insanitary conditions with poor access to feed
supplies may predispose sheep to listeriosis in intensive farms.
2. Pathogen risk factors
Factors that increase the infection pressure largely involve a massive
multiplication of L. monocytogenes in the feed or environment.
For example the organism can multiply in silages, poorly preserved
hay and grain and can exist in environment even at law temperature.
That is why feeding of grass or corn silage is considered as a major
risk factor for the occurrence of listeriosis.
The increase in use of silage for feed in ruminants may be the reason
587
 List. cont’d---
for the apparent increase in the prevalence of the disease in
intensive farms.
Other pathogenic risk factor is the ability of the organism to
persist in faces, straw, silage and in soil as well as in a low
temperature for a long time.
For example the organism persists for as long as 3 months in
sheep feces and has been shown to survive for up to 11.5 months
in damp soil to 207 days on dry straw and for more than 2 years
in dry soil and feces.
It is resistant to temperatures of - 20°C for 2 years.
Pathogenesis
You know that, the natural habitat of Listeria species is soil.
Plants decaying vegetation and silage(PH over 5.5) are in which
the bacteria can multiply.
Asymptomatic faecal carriers occur in man and many animals.
Silage is commonly implicated in outbreak of listeriosis in cattle
588
 List. cont’d---
and sheep.
Ingestion of the organism with penetration of the mucosa of the
intestine is the most common portal of entery of the organism.
The pathogenic species of Listeria infect the organisms through
1. The epithelium of small intestine( by damaging it)
The organism is a facultative intracellular pathogen that can infect
cells including intestinal cells by directed endocytosis.
It can survive and overcome the hepatic and splenic macrophages
by
 The bacterial superoxide dismutase that protect against the
bactericidal activity of the respiratory burst of the phagocyte.
Listerolysin produced by L. monocytogenes disrupts lysosomal
membranes allowing the organism to grow in the cytoplasm.
2. The damaged mucosal surface to central nervous system, via
neural sheath of the nerve endings of the trigeminal nerve.
Portal of entry is by ascending infection of the trigeminal or other
cranial nerves following loss of the integrity of the buccal mucosa
589
 List. cont’d---
resulting from trauma, the shedding of deciduous or permanent
teeth or from periodontitis.
The bacteria can infect the organism through the sensory nerves
of the skin following dermatitis from prolonged wetting of the
fleece to spinal cord and cause spinal myelitis.
Clinical findings
When disease occurs it is usual to have an outbreak of different
forms.
The most common clinical manifestation of listeriosis are
1. Septicemic listeriosis
2. Listerial encephalitis
3. Listerial abortion
4. Spinal myelitis
5. Ophthalmitis and iritis
6. Listerial mastitis
590
 List. cont’d---
1.Septicemic listeriosis
Septicemic form of listeriosis is common in newborn lambs, calves
and foal.
Rarely it occurs in ewes and goats after listerial abortion if the fetus
is retained.
There are no signs suggestive of nervous system involvement.
The syndrome being a general one in septicemic form of listeriosis
comprising
Depression
Weakness and emaciation
 Pyrexia and diarrhea
In some cases with hepatic necrosis and gastroenteritis at necropsy.
Corneal opacity is accompanied by dyspnea, nystagmus and mild
opisthotonos.
2. Listerial encephalitis
Fever usually reaches 40°C but occasionally as high as 42°C at
early stage. 591
 List. cont’d---
However in the later stage the body temperature becomes normal.
During the early stage the sheep may show a frenetic desire( fast
and energetic as wild) to escape.
But are uncoordinated as they run and fall easily.
The syndrome progresses rapidly with more severe depression to
the point of somnolence and the development of signs of cranial
nerve dysfunction.
After this the following clinical signs are developed
Incoordination
Head deviation sometimes with head tilt
Walking in circles
Unilateral facial hypalgesia and facial paralysis
Facial hypalgesia( diminished sensitivity to pain) can be detected
with pressure from a haemostatic forceps.
While the facial paralysis is manifest with
Drooping of the ear
Paralysis of the lips 592
Head deviation with head tilt of sheep due to listerial encephalitis

593
594
595
596
597
598
599
600
Paresis of the muscles of the jaw

601
Neural form listeriosis in calves
Iritis in cattle due to listeriosis

602
 List. cont’d---
Ptosis (dropping of the lower eyelid/ s)on the same side of the face
as the hypalgesia.
This may be accompanied by exposure keratitis often severe enough
to cause corneal ulceration.
 Strabismus (improperly aligned eyes) and nystagmus( involuntary
eye movement) occur in some affected animals.
Panophthalmitis with pus evident in the anterior chamber of one or
both eyes,
Also there is paresis of the muscles of the jaw with poor tone or a
dropped jaw.
As the result prehension and mastication are slow and the animal
may stand for long periods drooling saliva and with food hanging
from its mouth.
3. Listerial abortion
Outbreaks of abortion because of listeria organism are more
common in sheep and in goats.
In sheep and goats abortions occur from 3 months of pregnancy 603
 List. cont’d---
onwards.
Afterbirth is usually retained and there is a blood stained vaginal
discharge for several days.
There may be some deaths of ewes from septicemia if the fetus is
retained.
Abortion because of listeria occurs also in cattle and rarely in pigs.
In cattie, abortion or stillbirth occurs sporadically and usually in the
last third( trimester) of pregnancy.
Retention of the afterbirth occurs commonly with clinical illness and
fever of upto 40. 5oC.
Abortion has been observed soon after the commencement of silage
feeding but not always.
4. Spinal myelitis
Affected animals are initially mentally alert, bright and continue
to eat.
There is fever, ataxia with initial knuckling of the hind limbs.
Then after there is weakness and paralysis of hind limbs. 604
 List. cont’d---
In some cases, both in sheep and cattle, there is also paresis and
paralysis of the front limbs.
Lastly there is rapid deterioration of affected animals.
So the affected animals are commonly humanely killed.
5. Ophthalmitis and iritis
There is swelling of the iris and constriction of the pupil.
White focal lesions are evident on the internal surface of the cornea
with floccular material in the anterior chamber.
In advanced cases have pannus and corneal opacity. One or both
eyes are affected.
6. Listerial mastitis
Infection in the udder may involve in a single quarter or in both
quarters .
It is chronic and has poor responsive to treatment.
There is a high somatic cell count in milk from the affected quarter.
But the milk appears physically normal.
605
 List. cont’d---
Necropsy findings
Typically there are no distinctive gross changes associated with
listerial encephalitis.
Histological examination of central nervous system tissue is
necessary to demonstrate the microabscesses that are characteristic
of the disease.
The microabscesses are present in the brainstem in listerial
encephalitis and in the cervical and/or lumbar spinal cord in
outbreaks of spinal myelitis.
Visceral lesions occur as multiple foci of necrosis in the liver,
spleen, and myocardium in the septicemic form and in aborted
fetuses.
Aborted fetuses are usually edematous and autolyzed, with very
large numbers of bacteria visible microscopically in a variety of
tissues.
In aborting dams, there is placentitis and endometritis in addition to
606
the lesions in the fetus.
Diagnosis
Diagnose of listeriosis can be done based on
Epidemiology of the disease
Clinical findings
Necropsy findings
 bacteriological
Serological
Confirmatory diagnose of listeriosis is done by isolation and
identification of the organism in the laboratory.
Specimens of choice depends on the form of listeriosis.
1. In visceral form of listeriosis the specimens of choice are
material from lesions of visceral organs like heart, lung, liver,
kidney or spleen.
2. In neural form of listeriosis the specimens of choice are
 Samples of lumbosacral CSF that can be collected under local
anesthesia.
 In cases of listeriosis, the CSF has an increased protein
concentration (0.6–2.0 gm/l [normal 0.3 gm/l]) and a mild 607
 List. cont’d---
pleocytosis composed of large mononuclear cells.
The cerebral spinal fluid(CSF) and half of midsagittally
sectioned brain including brainstem can be used for culturing and for FAT.
3. In abortion case of listeriosis
The specimens of choice are chilled placenta( cotyledons), fetal
abomasal contents and/ or uterine discharges and/ or vaginal discharges
for culturing and FAT.
4. In listerial mastitis case
Mastitic milk is the specimens of choice.
NB. If the specimens are from neural cases of listeriosis a cold –
enrichment is necessary to isolate the bacteria.
NB. Parallel collect the above tissue samples and preserve with buffer
formalin for histopathology.
Serological tests (agglutination and complement fixation tests) have been
used.
But they lack the predictive value and not widely used.
Differential diagnosis
Listeriosis must be differentiated from 608
 List. cont’d---
1. Listerial encephalitis must be differentiated from
Pregnancy toxemia in sheep( pregnant lipidosis)
Nervous ketosis in cattle
Rabies
Gid( Coenuruses cerebrals)
Polioencephalomalacia
Scrapie (transmissible spongiform encephalopathy(TSE) in sheep
and bovine spongiform encephalopathy(BSE)
2. Listerial abortion must be differentiated from
Brucellosis
Listeriosis
Campylobacteriosis
Leptospirosis
Trichomoniasis
Mycosis
Epizootic bovine abortion
609
 List. cont’d---
BVD(Bovine virus diarrhoea
3. Listerial iritis must be differentiated from
Infectious Bovine Keratoconjuctivitis(IBK)
Treatment
Recovery from listeriosis depends on early aggressive antibiotic
treatment.
If signs of encephalitis are severe, death usually occurs despite
treatment.
L. monocytogenes is susceptible to
 Penicillin (the drug of choice)
Ceftiofur
 Erythromycin
For sulpha drugs like trimethoprim/sulfonamide
Chlortetracycline
In visceral and abortion case of listeriosis, administer the above
antimicrobial drugs systemically.
High doses are required to treat listerial encephalitis. 610
 List. cont’d---
In visceral and abortion case of listeriosis, administer the above
antimicrobial drugs systemically.
High doses are required to treat listerial encephalitis.
Because of the difficulty in achieving minimum bactericidal
concentrations in the brain.
So that Penicillin G should be given at 44,000 U/kg BW,IM daily for
1–2 week.
The first injection should be accompanied by the same dose given
IV.
Intravenous injection of chlortetracycline (10 mg/kg BW per day for
5 days) is reasonably effective in meningoencephalitis of cattle
Supportive therapy including fluids and electrolytes is required for
animals having difficulty eating and drinking.
Treatment of listerial iritis is with systemic administration of the
above antibiotics in the early stages coupled with subpalpebral
administration corticosteroid and atropine sulphate to dilate the
pupil. 611
 List. cont’d---
Prevention and control measures
Listeriosis prevention and control measures is difficult because of
The ubiquitous occurrence of the organism
 The lack of a simple method of determining when it is present in
high numbers in the environment.
 Poor understanding of the risk factors other than silage.
So that where the risk factor is silage use of the particular silage
should be discontinued on a trial basis when there is listeriosis
outbreak.
Generally spoiled silage should be avoided.
In feedlot intensive farms, tetracyclines is used in prophylactic dose
to prevent listeriosis.
Other recommendations on the feeding of silage include:
Avoiding making silage from fields where molehills may have
contaminated the grass.
 Avoiding soil contamination when filling the clamp
Corn ensiled before being too mature and grass silage containing612
 List. cont’d---
additives are likely to have a more acid PH which discourages
multiplication of L. monocytogenes.
Silage removed from the clamp should be fed as soon as possible.
Where iritis is a problem, feeding systems that avoid eye contact
with silage should be used.
Use of vaccines have been equivocal with the sporadic nature of the
disease lead to questions about the cost-benefit of vaccination.
In European countries A live attenuated vaccine has been
shown to induce protection against intravenous challenge.
The vaccine reduce the annual incidence of listeriosis in sheep
from 4% to 1.5% .
Commercial killed vaccines are available for the control of the
disease.
The efficacy of vaccination still requires further determination.
But when economy or food availability on the farm dictate that
contaminated silage must be fed.
So some consideration might be given to vaccination as a means of
613
providing some protection.
Diseases associated with Erysipelothrix rhusiopathiae
(insidiosa)
16. Swine Erysipelas
Erysipelas is an infectious disease and is one of the oldest known
swine disease that affect growing and adult swine.
Acute outbreaks are characterized by
Sudden and unexpected deaths
 Febrile episodes
 Painful joints
Skin lesions that vary from generalized cyanosis to the often-
described diamond skin (rhomboid urticaria) lesions.
Chronic erysipelas tends to follow acute outbreaks and is
characterized by
Low mortality
Enlarged joints
Lameness
Postmortem evidence of vegetative endocarditis. 614
 Ery. cont’d---
Pigs with valvular lesions may exhibit few clinical signs.
However, when exerted physically they may show signs of
respiratory distress and possibly succumb to the infection.
It has zoonotic implication because it can affect human beings.
Erysipelas affects
 Mostly veterinarians(exposed to infection when vaccinating
with virulent with virulent culture).
Abattoir workers or butchers or those employed in similar trades.
It usually produces a swollen finger and is known as erysipeloid.
Etiology
Erysipelas is an infectious disease caused by Erysipelothrix
rhusiopathiae.
The organism is Gram positive rod widely distributed in animals and
in the environment.
E. rhusiopathiae can grow on non-enriched media produces
pinpoint, non hemolytic colonies after incubation for 24 hr.
After 48 hr of incubation a zone of incomplete hemolysis becomes 615
 Ery. cont’d---
The main diseases of domestic animals caused by Erysipelothrix rhusipathiae.
Main host(hosts) Disease / Disease syndrome
Swine erysipelas
Acute septicaemic form( pregnant sows may
abort)
Pigs Urticarial form(diamond skin disease)
Endocarditis (chronic form)
Polyarthritis(chronic form)
Acute septicemia
Turkeys, geese and other birds Endocarditis(chronic form)
Arthritis(chronic)
Cattle, horse, rabbit and dog Occasional infection of varying severity
Young sheep Poly arthritis( chronic)
Adult sheep Lameness
Humans Erysipeloid and rarely endocarditis and arthritis

616
 Ery. cont’d---
evident around colonies.
It able to grow in temperature range 5 – 420C, with in PH range 6.7
– 9.2
It grows in common media like nutrient agar.
But the growth is improved by addition of serum or blood.
It doesn’t grow on MacConkey agar.
Up to 50% of pigs in intensive swine production areas are
considered to be colonized with E. rhusiopathiae.
The organism commonly resides in the tonsillar tissue.
At least 32 different serotypes are recognized, and pigs are
considered to be susceptible to at least 15 of them.
Many of the non pathogenic serotypes have been regrouped and
called Erysipelothrix tonsillarum.
These are the nonpathogenic type found in the tonsil and are
morphologically and biochemically similar to E. rhusiopathiae.
But has a very distinctive genetic profile.
617
 Ery. cont’d---
Epidemiology
Erysipelas in pigs occurs worldwide.
In most countries has reached a level of incidence sufficient to cause
serious economic loss due to
Deaths of pigs and
Devaluation of pig carcasses because of arthritis.
Total indoor confinement of swine herds(in intensive swine farms)
make the pigs not to have contact with contaminated soil.
As the result the occurrence of the disease has decreased markedly
in intensive farms.
The exception to this would be outdoor units( extensive and semi -
intensive swine farms) where no vaccination is practiced.
The disease occurred most commonly in unvaccinated growing pigs
over 3 months of age and adults.
This is primarily because maternal-derived antibodies provide
passive immunity and believed to last up to 3 months.
618
 Ery. cont’d---
While older pigs tend to develop protective active immunity as a
result of exposure to serotypes that do not induce clinical disease.
Erysipelas infection in pigs is usually caused by serotypes 1 or 2.
These serotypes has also been demonstrated in wild boars.
So that wild boars should not be ignored in epidemiology erysipelas
since they act as a reservoir.
Morbidity and case fatality rates in pigs vary considerably from area
to area.
Because of variations in virulence of the particular strain of the
organism involved in infection.
In unvaccinated pigs, the morbidity in the acute form will vary from
10 - 30%; the case fatality rate may be as high as 75 % .
Source of infection
Source of infection can be
Sick animals
Carriers
Prevalence of infection with E. rhusiopathiae in pigs in most 619
 Ery. cont’d---
surveys indicating that 20 - 40% of pigs are carriers.
This explains why the usual source of infection are carrier animals.
Methods of transmission
Ingestion from contaminated soil
Mechanical vectors ( house fly)
Skin abrasions
NB. The environment is considered to be secondary to animals as a
reservoir of infection.
The organism has been isolated from a horse affected with
vegetative endocarditis .
It is possible that other species, such as cattle may harbor strains
that are pathogenic for swine.
Risk factors
Factors that affect the occurrence of erysipelas infection are
1. Animal risk factors
2. Pathogen risk factors
620
 Ery. cont’d---
1. Animal risk factors
Pigs of all ages are susceptible to infection.
Although adult pigs are most likely to be affected if the local strain
is relatively low virulence.
When the strain is virulent, pigs of all ages, even sucklers a few
weeks old, develop the disease.
Recently farrowed sows seem to be particularly susceptible.
This suggests that fatigue may be a factor.
There are many additional predisposing factors.
Some of them are
Sudden diet change
Over stocking density
Heat and cold stress
Concurrent viral infection especially hog cholera may increase
susceptibility of the host.
It is likely that the animals are immune to the strains that are
normally found in their particular environment. 621
 Ery. cont’d---
But erysipelas outbreak may occur when there is
Arrival of new serotypes through new pig arrivals or
 The turning over of previously contaminated land together with an
increase in stress that are the main factors for the occurrence of
erysipelas.
2. Pathogen risk factor
At least 32 serotypes of Erysipelothrix rhusiopathiae are known to
exist.
However only 15 of them are commonly affecting pigs.
Among the 15 serotypes, serotypes 1 and 2 are the most common
serotypes isolated from swine affected with clinical erysipelas.
So that serotypes 1 and 2 are generally believed to be the only
serotypes that cause the acute erysipelas.
The other serotypes are relatively uncommon and none of them has
yet been a cause of acute erysipelas epidemics.
Pathogenesis
The bacteria infect susceptible animals mainly through scarified 622
 Ery. cont’d---
wound.
The virulence factors for pathogenic serotypes are
Their ability to produce a capsule that resists phagocytosis.
Some others may produce a neuraminidase which may cleave the
mucopolysaccharides in cell walls and cause vascular damage
leading to hemorrhage and thrombosis.
Coagulase activity is a possible virulence factor of the organism
In the first instance invasion of the bloodstream occurs in all
infected animals.
The subsequent development of either an acute septicemia or a
bacteremia with localization in organs and joints is depend on
undetermined factors.
Virulence of the particular strain may be important and this may
depend upon the number of recent pig passages experienced.
Localization in the chronic form is commonly in the skin, joints and
on other heart valves, with probable subsequent bacteremic episodes.
Selective adherence of some strains of E. rhusiopathiae (insidiosa)
623
 Ery. cont’d---
to heart valves may be a factor in the pathogenesis of endocarditis.
In joints, the initial lesion is an increase in synovial fluid and
hyperemia of the synovial membrane.
And followed in several weeks by the proliferation of synovial villi
(really a synovitis)
Then thickening of the joint capsule and enlargement of the local
lymph nodes
Amyloidosis may occur in pigs with chronic erysipelas
polyarthritis.
The microscopic lesions include vasculitis in capillaries and venules
in many sites, including glomeruli, pulmonary capillaries and the
skin.
Clinical findings
Depending on the length of incubation period, erysipelas in pigs has
the following forms
1. Peracute
2. Acute 624
 Ery. cont’d---
3. Chronic
By localization of pathological process erysipelas can be
1. Septicemic form of erysipelas occur mostly in peracute and acute
form the disease
2. Skin form also called urticarial form(diamond skin disease)
occurs mostly in acute form of erysipelas.
 3. Endocarditis and polyarthritis form of erysipelas occurs mostly
in chronic form of erysipelas.
1. Peracute form of erysipelas
It is common in young pigs and has septicemic form.
It is not in adult pigs and characterized by
 Sudden death or
Dullness and depression with increased body temperature upto 420C.
2. Acute form of erysipelas
Acute septicemic form of erysipelas has an incubation period of 1 -
7 days with clinical signs of
625
 Ery. cont’d---
Sudden onset of high fever upto 42°C
Followed sometime later by severe prostration, complete anorexia,
thirst and occasional vomiting.
Initially, affected pigs may be quite active and continue to eat even
though their body temperature is high.
Severely affected pigs showing marked red to purple discoloration
of the skin of the jowl and ventral surface or the whole-body may be
cyanotic.
Some pigs may be reluctance to rise and some incoordination while
walking.
Dyspnea is a common feature in erysipelas of pigs
Conjunctivitis with ocular discharge may be present.
Skin lesions are almost the pathognomonic signs in swine
erysipelas.
However it may not always be apparent.
These may take the form of the classical diamond shaped, red
urticarial plaques about 2.5 – 5 cm2. or 626
Typical rhomboid urticarial lesions (“diamond-skin” lesions) of
swine erysipelas

627
Diamond shaped and red urticarial plaques on the skin of swines

628
629
Diamond shaped and red urticarial plaques on the skin of swines

630
631
 Ery. cont’d---
Amore diffuse edematous eruption with the same appearance may
appear.
The lesions are most common on the belly, inside the thighs and on
the throat, neck and ears.
Sometimes they can be felt rather than seen.
After a course of 2 - 4 days the pig recovers or dies, with diarrhea,
dyspnea, and cyanosis evident terminally.
The mortality rate may reach 75 % but wide variation may occur.
Pregnant animals may abort and it is thought that this is due to the
fever.
But it may be that there is a direct fetal action as congenital
infections and isolations of the organism from the fetus have
occurred.
The so-called 'skin' form is usually the acute form with more
prominent skin localization.
But less severe signs of septicemia and with a low mortality.
632
 Ery. cont’d---
The skin lesions disappear in about 10 days without residual effects.
In the more serious cases the plaques spread and coalesce, often
over the back, to form a continuous, deep purple area extending
over a greater part of the skin surface.
The affected skin becomes black and hard, and the edges curl up
and separate from an underlying raw surface.
The dry skin may hang on for a considerable time and rattle while
the pig walks, or it may slough off.
Chronic form of erysipelas
Chronic form of erysipelas is characterized by arthritis and
endocarditis.
1. In arthritis form, signs are vague and indistinct except for the
joint lesions which is characteristic of this form of the disease.
Joint lesions are found in elbow, hip, hock, stifle and knee joints
and cause lameness and stiffness.
The joints are obviously enlarged and are usually hot and painful at
first. 633
Coalesced plaques in serious form of erysipelas

634
Slough off the skin affected by erysipelas.

635
636
 Ery. cont’d---
The skin lesions disappear in about 10 days without residual effects.
In the more serious cases the plaques spread and coalesce, often over
the back, to form a continuous, deep purple area extending over a
greater part of the skin surface.
The affected skin becomes black and hard, and the edges curl up and
separate from an underlying raw surface.
The dry skin may hang on for a considerable time and rattle while
the pig walks, or it may slough off.
Chronic form of erysipelas
Chronic form of erysipelas is characterized by arthritis and
endocarditis.
In arthritis form, signs are vague and indistinct except for the joint
lesions which is characteristic of this form of the disease.
Joint lesions are found in elbow, hip, hock, stifle and knee joints and
cause lameness and stiffness.
The joints are obviously enlarged and are usually hot and painful at
first. 637
Alopecia and sloughing of the tail and tips of the ears.

638
 Ery. cont’d---
In endocarditis form aloud murmur sound which is audible on
auscultation if the valves are badly damaged.
This result in dyspnea and cyanosis and even upto sudden death.
Paraplegia in pigs during erysipelas infection

639
Crippling, irreversible arthritis in a growing pig

640
Erysipelas is zoonotic

641
 Ery. cont’d---
Necropsy findings
In the peracute cases all that may be seen is a congested carcass
with discoloration of the skin.
Classic 'diamond skin' lesions may be present.
However, the diffuse, purplish discoloration of the belly and
cyanosis of the extremities is more common in erysipelas than
other septicemic diseases of pigs.
It is a more reliable histopathological finding during erysipelas.
Internally, petechial and ecchymotic hemorrhage occurs mainly
on the pleura and peritoneum and beneath the renal capsule.
Petechial and ecchymotic hemorrhage also found on the heart,
liver and spleen.
Mesenteric lymph nodes are swollen and hemorrhagic.
There is congestion of the lungs and liver.
Infarcts may be present in the spleen and kidney.
642
Carcass of swine affected by erysipelas

643
644
 Ery. cont’d---
In chronic form of erysipelas the necropsy findings are found mainly
in joints and endocardium of heart.
In arthritis form, there is a non suppurative proliferative arthritis
involving limb and intervertebral joints.
A synovitis, with a serous or sero fibrinous amber-colored intra-
articular effusion is common.
Local lymph node enlargement is usual.
With time, the joint lesions often repair by fibrosis and ankylosis
sufficiently to permit use of the limb.
Endocardial lesions when present are large, friable vegetations on
the valves often of have sufficient size to block the valvular orifice
and cause permanent valvular defect.
Diagnose
Diagnosis of erysipelas is done based on
Epidemiology
Clinical sign
645
Necropsy findings
Valvular defect caused endocardial lesions due to erysipelas

646
 Ery. cont’d---
Bacteriological
Serology
Molecular
Bacteriological diagnose starts with th examination of blood
smears in acute case.
Examination of blood smears may reveal the presence of the
bacteria, particularly in the leukocytes.
But blood culture is likely to be more successful as a method of
diagnosis in acute form of erysipelas.
Culture swabs from joints, synovial membranes in culture media,
heart valve masses, spleen, kidney, and bone marrow, particularly
from a long bone.
Recovery of the bacterium from skin lesions and from chronic
forms of a disease may be difficult.
Serology
The commonest serological tests that are used to diagnose
647
erysipelas are
 Ery. cont’d---
1. Florescent techniques(FAT) have been developed to show
antigen in joints.
2. Serum agglutination tests(SAT) used in herd level because it has
low predictive value.
Efficient serological tests that are used to diagnose individual
affected(carrier) animals are
3. CFT and ELISA
Differential diagnose
Erysipelas in pigs is not ordinarily difficult to diagnose,
Because of the characteristic clinical and necropsy findings.
The acute disease may be confused with the other septicemias
affecting pigs.
But pigs with erysipelas usually show the characteristic skin
lesions(diamond shaped and red urticarial plaques).
So that erysipelas must be differentiated from
1. Septicemic salmonellosis which is characterized by
Gross blue-purplish discoloration of the skin especially the ears 648
 Ery. cont’d---
 Some evidence of enteritis and polypnea and dyspnea.
2. Hog cholera(classical swine fever) which is characterized by
Affecting large numbers of pigs quickly than erysipelas
Weakness and fever
 Muscle tremors, skin discoloration and rapid death
Convulsions are also common
3. Streptococcal septicemia and arthritis
They are almost entirely confined to suckling pigs in the first few
weeks of life.
Streptococcal septicemia is usually associated with Actinobacillus
suis.
4. Streptococcal endocarditis
It causes endocarditis in similar age as erysipelas
So that bacteriological examination is necessary to differentiate
them.
5. Mycoplasma arthritis
Generally it affects pigs less than 1 0 weeks of age and produces649 a
 Ery. cont’d---
polyserositis as well as polyarthritis.
However, Mycoplasma hyosynoviae can produce simple
polyarthritis in growing pigs.
Arthritis is not sever as compared with arthritis because of
erysipelas.
However cultural differentiation is frequently necessary.
6. Rickets and chronic zinc poisoning
They produce lameness in pigs but they occur under special
circumstances.
They are not associated with fever.
Rickets is accompanied by abnormalities of posture and gait that
are not seen in erysipelas
7. Foot rot of pigs
It is easily differentiated by the swelling of the hoof and the
development of discharging sinuses at the coronet.
Treatment
Penicillin and anti-erysipelas serum comprise the standard 650
 Ery. cont’d---
treatment.
They are often administered together by dissolving the penicillin
in the serum.
Penicillin alone is usually adequate when the strain has only mild
virulence.
Standard dose rates give a good response in the field.
But experimental studies suggest that 50 000 IU/kg BW of procaine
penicillin intramuscularly for 3 days are required for complete
chemotherapeutic effect.
Oxytetracycline is also effective in erysipelas.
Chronic cases( arthritis and endocarditis) do not respond well to
either treatment.
This is because of
The structural damage that occurs in joints and
 The inaccessibility of the organism in the endocardial lesions.
Most strains are resistant to apramycin, neomycin, streptomycin
and also sulfonamides and polymyxins. 651
 Ery. cont’d---
Prevention and control measures
Successful prevention and control of erysipelas depends on
Good hygienic condition
Biosecurity (other pigs and other species)
Quarantine
Reduction of stress
An effective 6-monthly vaccination policy, preferably two doses, for
all animals including boars over 3 months of age.
 Rapid diagnosis and treatment.
1. General hygienic precautions
Clinically affected animals should be disposed of quickly
All new introduced animals should be isolated and examined for
signs of arthritis and endocarditis.
Eradication is virtually impossible.
Because of the ubiquitous nature of the organism; as the organism is
resistance to adverse environmental conditions.
Complete removal of all pigs and leaving the pens unstocked and652
 Ery. cont’d---
cleaning and disinfection is seldom satisfactory.
All animals dying of the disease should be properly incinerated to
avoid contamination of the environment.
Cleaning of the premises and the use of very strong disinfectant
solutions like 2% caustic soda(NaOH) and hypochlorites with 5%
active chlorine is advisable.
Immunization with biological agents
Biological prophylactic agents are commonly used to prevent and
control erysipelas.
The immunizing agents that are available include
1. Hyperimmune serum called anti - erysipelas serum
2. Vaccines
The parenteral administration of 5 - 20 ml of serum( amount
depending on age) will protect in contact pigs for a minimum of 1 -
2 weeks.
Suckling pigs in herds where the disease is endemic should receive
10 ml of hyperimmune serum during the first week of life and at653
 Ery. cont’d---
monthly intervals until they are actively vaccinated.
Active immunization of pigs can be done as early as 6 weeks
provided the sows have not been vaccinated.
However, in herds where sows are routinely vaccinated prior to
farrowing, persisting maternal passive immunity may require that
piglet vaccination be delayed until 10 - 12 weeks of age for effective
active immunity.
NB. Because of anaphylaxis which is happened due to repeated
administration of serum it is not widely used.
There is no fully satisfactory vaccine available for erysipelas
because of
 Strain variation and
 Short duration of immunity
However, the vaccines may reduce the occurrence of clinical
disease.
So that regular administration of vaccine with 6-months interval
overcomes short duration of the immunity to some extent. 654
 Ery. cont’d---
But there is always the possibility of appearing of a new strain.
Following a single vaccination at 6 - 10 weeks of age, significantly
protects the young pigs upto market age.
However, a second 'booster‘ vaccination given 2 - 4 weeks later is
advisable.
There are two types of vaccines in developed countries
Inactivated ( bacterin) vaccine
Live attenuated vaccine
Inactivated vaccine and live vaccine provide immunity almost for 6
and 8 -10 months respectively.
Sows should be vaccinated twice yearly preferably 3 - 6 weeks
before farrowing.
Because this will also provide significant protection against the
septicemic form of erysipelas in young sucklers.
Live attenuated vaccines may produce hemorrhage in the skin at the
injection site.
655
Diseases associated with Leptospira spp.
17. Leptospirosis
It is infectious bacterial disease of many domestic animals including
birds.
It can appear as clinical disease or in latent form.
Clinical leptospirosis is characterized by
Fever
Haemoglobinuria
 Icterus and necrosis of the mucous membrane and skin
Atony of GIT tract
Abortion or still birth
Leptospirosis affects human being.
So that it is zoonotic disease.
Etiology
Leptospira is found under the order of spirochaetales, family
Leptospiraceae and under genus Leptospira.
They are slender, motile, flexuous, unicellular, helically coiled 656
 Lepto cont’d---
ranging from 0.1 - 3µm in width.
The outer sheath is the outermost layer of a spirochaetes cell and is
a multilayered membrane.
The outer sheath completely surrounds the periplasmic
flagella(axial filament) and the helical protoplasmic cylinder.
The helical cylinder consists
 A. Nuclear material
 B. Cytoplasm
 C. Cytoplasmic membrane
 D. Peptidoglycan layer of cell wall.
 E. Periplasmic flagella
 F. The outer sheath
The periplasmic flagella are wrapped around the cylinder and are
located in the periplasmic space of these Gram negative bacteria
One end of each flagellum is inserted near a pole of the
protoplasmic cylinder and attached to a plate – like structure called
insertion discs. 657
Structural features order spirochates

658
Section through spirochates demonstrating structural features

659
 Lepto cont’d---
The other end of each flagella are not inserted and extends to the
center of the cell.
So each flagellum may over lap the others.
These periplasmic flagella facilitate the motility of bacteria in viscid
environment.
Leptospira are aerobic bacteria that are visible under dark field
microscopy, silver impregnation and FA technique.
Their movement is rotational around central axis, flexing,
translational and may have adulatory movement .
Leptospira has many spirals which are tight and fine.
Either both or one end is hooked.
It is gram negative aerobic bacterium.
The genus Leptospira at present has two species.
 1. The pathogenic species – Leptospira interrogans
 2. Saprophytic species Leptospira biflexa
660
Morphology of Leptospira

661
 Lepto. cont’d---
The pathogenic Leptospira interrogans containing over 212
serovars arranged into 23 serogroups.
Differentiation between serovars belonging to a particular serogroup
is done by
Cross agglutination tests
Restriction endonuclease analysis (REA) of leptospiral DNA
Serological typing has revealed that there are many
serotypes(serovars)under which share antigenic configuration in
genus Leptospira.
Serovar( serotype) is the serologically least divisible recognized
type.
A group of serovars with common antigens is called a serogroup.
and are used for diagnostic convenience.
Serovars of L. interrogans that can cause disease in animals and
human beings vary between continents, countries and even in
regions of one country.
662
 Lepto. cont’d---
The media used to isolate leptospira are
Korthof
Stuarts broths Fletcher semi – solid medium
EMJH(Ellinghausen-McCullough-Johnson-Harris)
Oleic albumin complex (OAC) medium
All of them are liquid media.
But they can be made semi – solid media by addition of 1.5 gm agar/
liter of the medium.
Leptospira interrogans serovars can not be resolved under bright
field microscopy.
They stain weak or not at all with aniline and Romanowsky stains.
But leptospires can be demonstrated in urine and other body fluids
and tissues by dark field microscopy and Fluorescent Antibody
Techniques(FAT).

663
 Lepto. cont’d---
Common serovars of genus Leptospira and their reservoir hosts
Serovars Main reservoir(s) Alternative
reservoir(s)
L. interrogans serovar Wild life Has no
autominalis
L. interrogans serovar ballum Wild life Has no
L. interrogans serovar Pigs, horse and cattle Has no
bratislava
L. interrogans serovar canicola Dogs Cattle, pigs and
rodents
L. interrogans serovar Wild life Dogs, cattle and pigs
grippotyphosa
L. interrogans serovar hardjo Cattle Sheep
L. interrogans serovar ictero - Rats Dogs ,cattle and pigs
haemorrhagiae
L. interrogans serovar pomona Pigs and cattle Dogs and wild life

664
Epidemiology
The disease has world wide distribution.
However, most leptospiral infections are subclinical and infection is
more common than clinical disease.
L. interrogans serovar pomona is the commonest serovar of
Leptospira that cause infection in all farm animals.
But its international distribution is unpredictable.
Serological surveys of cattle in the African continent reveal
evidence of antibodies against numerous leptospiral serovars.
In West Africa, serosurveys of dairy herds revealed 45% of cattle
were positive to one or more serovars.
This probably represented natural infection because vaccination had
not been practiced.
Leptospirosis affects all farm animal species.
The epidemiological characteristics of the infection, some of which
are unique with a species and important in the diagnosis, treatment
and control strategies.
665
 Lepto. cont’d---
The epidemiology of leptospirosis is most easily understood by
classifying the disease into two broad categories:
Host-adapted leptospirosis
 Non-host-adapted leptospirosis.
An animal infected with a host-adapted serovar of the organism
is called a 'maintenance' or 'reservoir' host.
Exposure of susceptible animals to non host-adapted serovars results
in accidental or incidental disease.
Each serovar is adapted to a particular maintenance host, although
they may cause disease in any mammalian species.
A serovar behaves differently within its maintenance host species
than it does in other, incidental or accidental hosts.
A maintenance host is characterized by
A high susceptibility to infection
Endemic transmission within the host species
 Relatively low pathogenicity for its host
A tendency to cause chronic rather than acute disease 666
 Lepto. cont’d---
Persistence of the serovar in the kidney and sometimes the genital
tract
A low antibody response to infection with difficulties in diagnosis
Low efficacy of vaccination in prevention of infection
In contrast an incidental host is characterized by
Relatively low susceptibility to infection but high pathogenicity for
the host
 A tendency to cause acute, severe rather than chronic disease
 Sporadic transmission within the host species and acquisition of
infection from other species, sometimes in epidemic form
 A short kidney phase
 A marked antibody response to infection, making for ease of
diagnosis
 More efficacious vaccines for preventing infection.
Cattle are the maintenance host and reservoir for L. interrogans
serovar hardjo.
It is an important cause of bovine abortion and is the commonest667
 Lepto. cont’d---
Some common leptospiral serovars with their maintenance and
accidental hosts
Serovar Maintenance host Accidental host
L. interrogans serovar hardjo Cattle and pig Sheep and man

L. interrogans serovar Pig, skunk, raccoon and Sheep and cattle

pomona opossum
L. interrogans serovar Wild life(Raccoon, Sheep and cattle
grippotyphosa opossum and
Squirrel)
L. interrogans serovar ictero Brown rat Cattle and pig
- haemorrhagiae

668
669
 Lepto. cont’d---
serovar.
Leptospirosis in sheep and goats is caused by L. interrogans serovar
grippotyphosa.
Leptospirosis in sheep can cause lamb losses due to
Congenital infections.
Starvation of lambs because of the acute agalactia of ewes.
 Fatal infection of feedlot lambs
L. interrogans serovar pomona has been the predominant serovar in
leptospiral infection in pigs.
However, there are other most common serovars of leptospira in
which antibodies are found in pigs.
These are
L. interrogans serovar ictero - haemorrhagiae
L. interrogans serovar bratislava
L. interrogans serovar autumnalis
L. interrogans serovar pomona
The clinical signs often associated with these infections is poor 670
 Lepto. cont’d---
reproductive performance.
Seropositive sows have a greater risk of weak newborn pigs and
having more weak newborn piglets per litter.
House mice on swine farms may be serologically positive to L.
interrogans serovar bratislava.
Which suggests a possible source of the organism.
Introduction into a farm may be via an imported boar who
frequently is found to harbor leptospira in the genital tract.
Leptospirosis in horses may cause
1.Recurrent uveitis
2. Nonulcerative keratouveitis
3. Neonatal foal disease
4. Abortion and stillbirths
1.Recurrent uveitis
Leptospirosis is relatively mild in the horse, except for blindness due
to recurrent uveitis or periodic ophthalmia.
Leptospirosis with recurrent uveitis has higher prevalence in tropical
671
 Lepto. cont’d---
areas.
The dominant serovar varies widely between localities.
But L. interrogans serovar bratislava has higher seroprevalence in
horses.
This help us to suggest that, the horse may be a maintenance host for
this serovar.
2. Nonulcerative keratouveitis
Nonulcerative uveitis associated with leptospira infection is common
in 2-yearold horses.
The horse was seropositive to three serovars of L. interrogans
L. interrogans serovar pyrogens
L. interrogans serovar canicola
L. interrogans serovar ictero -haemorrhagiae
3. Neonatal foal disease
A severe rapidly fatal illness in foals characterized by
Massive pulmonary hemorrhage and
Kidney disease 672
 Lepto. cont’d---
Neonatal foal disease has been associated with serological evidence
of L. interrogans serovar bratislava.
4. Abortion and stillbirths
Leptospirosis is an important cause of abortions and stillbirths in
the horse.
The common serotypes that cause abortion and still birth in horses
those were isolated from aborted fetuses are
Leptospira interrogans serovar grippotyphosa
Leptospira interrogans serovar pomona
Leptospira interrogans serovar bratislava
NB. Leptospirosis has also been diagnosed as a cause of abortions,
stillbirths, neonatal illness and neonatal death on a horse farm
involved in a flooding incident.
Risk factors for occurrence of leptospirosis
There are many risk factors that are associated with occurrence of
leptospirosis.
The most common risk factors are 673
 Lepto. cont’d---
 1. Environmental and management risk factors
 2. Rodent, mice and wild life exposure
1. Environmental and management risk factors
Survival of the organism in the environment depends largely upon
variations in soil and water conditions in the contaminated area.
The organism is susceptible to drying and a PH lower than 6 or
greater than 8 is inhibitory.
Low urinary PH in cattle fed with brewer's grains may inactivate
leptospira in animals with leptospiruria.
Ambient temperatures lower than 7 - 10°C or higher than 34 - 36°C
are detrimental to its survival.
Ground surface moisture and water is the most important factor
governing the persistence of the organism in bedding or soil.
It can persist for as long as 183 days in water-saturated soil.
But it survives for only 30 min when the soil is air-dried.
It survives in free, surface water for long periods.
The survival period is longer in stagnant than in flowing water 674
 Lepto. cont’d---
although persistence in the latter for as long as 15 days has been
recorded.
Certain management factors have been identified which pose risks
of L. interrogans serovar hardjo infection being introduced into
dairy herds.
 Purchase of infected cattle
Co-grazing or common grazing with infected cattle or sheep
Purchase or loan of an infected bull
Access of cattle to contaminated water supplies such as streams,
rivers, flood or drainage water.
2. Rodent, mice and wild life exposure
Rodent and mice exposure of domestic animals is associated with
risk of exposure to all serovars of leptospira infection.
The wildlife index value and their contact with domestic animals
has risk of exposure to many serotypes of Leptospira.
Source of infection
Source of infection can be 675
 Lepto. cont’d---
Sick animals including wild life
Carrier domestic and wild life (latent infection)
 Mice and rodents
Methods of transmission
The source of infection is an infected animal and carriers which
contaminates pasture, drinking water and feed by
Infective urine
Aborted fetuses
Uterine discharges
A viable infected neonate can harbor the infection for several weeks
after birth.
The semen of an infected bull may contain leptospira and
transmission by natural breeding or artificial insemination can occur
but is uncommon.
In rams, the semen is likely to be infective for only a few days
during the period of leptospiraemia.
In boars there is no evidence of coital transmission. 676
 Lepto. cont’d---
The important factors that play crucial role in epidemiology of
leptospirosis are
Leptospiruria
Wildlife
Urine is the chief source of contamination.
All animals which have recovered from infection may intermittently
shed organisms in the urine.
These animals are carrier animals.
NB. In any species, the leptospira may persist in the kidney for much
longer periods than they can be recovered from the urine by routine
laboratory methods.
Wildlife can act as a source of leptospiral infection for domestic
animals
Many study shows that there is variable rates of seroprevalence to
leptospira in many domestic animals.
Some of wild animals that act as a source leptospiral infection are
White-tailed deer 677
 Lepto. cont’d---
Mule deer
Pronghorns
Moose
 Red deer and elk
The entrance of the organism into the body of susceptible animals
occur most probably through cutaneous
Transplacental transmission is uncommon.
But neonatal infection in uterus has occurred during delivery.
Economic importance
Leptospirosis is a major cause of economic loss in farm animals.
The majority of leptospiral infections are subclinical and associated
with fetal infections causing
Abortions
Stillbirths
It cause the birth of weak neonates with a high death rate in cattle,
sheep, horses and pigs.
In cattle leptospirosis cause epidemics of abortions, infertility and
678
 Lepto. cont’d---
increased culling rate.
Epidemics of agalactia in dairy herds called milk drop syndrome is
associated with infection with L. interrogans serovar hardjo.
Leptospirosis is zoonotic disease and has emerged as a globally
important infectious disease in human medicine.
Leptospirosis is zoonotic disease and has emerged as a globally
important infectious disease in human medicine.
It is uncommon in developed countries.
It is now recognized as an emerging, potentially epidemic disease
associated with excess rainfall in tropical settings.
That is why the incidence is increasing in developed countries which
is associated with travelers returning from endemic tropical
countries.
Leptospirosis is an important zoonosis and is an occupational hazard
to butchers, farmers and veterinarians.
Human infection is most likely to occur by contamination with
infected urine or uterine contents. 679
 Lepto. cont’d---
Veterinarians may become infected by handling the tissues and
urine of sows which have aborted from L. interrogans serovar
pomona infection.
Although leptospira may be present in cows’ milk for a few days at
the peak of fever in acute cases, the bacteria do not survive for long
in the milk and are destroyed by pasteurization.
Pathogenesis
Leptospires invade the host across mucosal surfaces or softened
skin.
They have the ability to bind to epithelial cells and attach to the
constituents of the extracellular matrix through an active process
involving surface proteins.
Pathogenic leptospires are found extracellular between cells of the
liver and kidney.
Release of cytokines such as Tumor Necrosis Factor - alpha
(TNF-α) from monocytes through the endotoxic activity of the
leptospira. 680
 Lepto. cont’d---
Induction of TNF- α release may damage the endothelial cells
which results in hemorrhage seen in severe leptospirosis.
Leptospirosis can occur as an acute and severe disease due to
Septicemia with evidence of endotoxemia such as hemorrhages,
hepatitis, nephritis and meningitis.
Subacute moderately severe disease with nephritis, hepatitis,
agalactia and meningitis. or
Chronic disease characterized by abortion, stillbirth and infertility.
Variations between serotypes of L. interrogans in their pathogenicity
also affect the nature of the signs which appear.
For example L. interrogans serovar pomona infections cause
 Intravascular hemolysis
 Interstitial nephritis
However, L. interrogans serovar hardjo does not produce
hamolysin and does not cause interstitial nephritis.
But it causes clinical infection in sexually mature, lactating or
pregnant females. 681
 Lepto. cont’d---
Induction of TNF- α release may damage the endothelial cells
which results in hemorrhage seen in severe leptospirosis.
Leptospirosis can occur as an acute and severe disease due to
Septicemia with evidence of endotoxemia such as hemorrhages,
hepatitis, nephritis and meningitis.
Subacute moderately severe disease with nephritis, hepatitis,
agalactia and meningitis. or
Chronic disease characterized by abortion, stillbirth and infertility.
Variations between serotypes of L. interrogans in their
pathogenicity also affect the nature of the signs which appear.
For example L. interrogans serovar pomona infections cause
 Intravascular hemolysis
 Interstitial nephritis
However, L. interrogans serovar hardjo does not produce
hamolysin and does not cause interstitial nephritis.
But it causes clinical infection in sexually mature, lactating or
pregnant females. 682
 Lepto. cont’d---
Forms of leptospirosis in animal species
Spp of animals Acute form Subacute Chronic form
form
Cattle + (Calves only) + +( Abortion)
Sheep and goat +(Includes abortion) - -

Pig + - +( Abortion)
(Rarely and only
Horse - + +
(Abortion and
periodic ophthalmia

683
 Lepto. cont’d---

Vascular injury also occurs in the kidney and if the haemolysis is

severe, there will be
Anaemia
 Anoxia and
 Haemoglobinuria nephrosis
And infection may localize in the renal parenchyma causing
Interstitial nephritis and
Persistence of the leptospira in these lesions results in prolonged
leptospiruria.
In the acute phase of the disease, the animal may die from
septicemia and/or hemolytic anemia and/or uremic or at both.
Following systemic invasion, abortion may occur due to fetal death,
with or without placental degeneration.
Abortion usually occurs several weeks after septicemia because of
the time required to produce the changes in the fetus, which is
684
 Lepto. cont’d---
usually autolyzed at birth.
Abortion occurs most commonly in the second half of pregnancy,
but may occur at any time from 4 months on.
Although abortion occurs commonly in both cattle and horses after
either the acute or subacute form of the disease, abortion without
prior clinical illness is also common.
This is particularly the case in sows and occurs to a less extent in
cows and mares; this may be due to degenerative changes in the
placental epithelium.
In acute form of leptospirosis, localization of leptospirae in nervous
tissue is common in sheep and goats and may result in the
appearance of signs of encephalitis.
In the subacute form, the pathogenesis is similar to that of the acute
septicemic form, except that the reaction is less severe.
It occurs in all species, but is the common form in adult cattle and
horses.
In this form of leptospirosis periodic ophthalmia (recurrent uveitis)
685
 Lepto. cont’d---
in horses and rarely in cattle with absence of signs of clinical
illness is common.
However, there is rising of antibody titers.
Clinical findings
In all animals the incubation period is 3 - 7 days.
The clinical findings in leptospirosis are similar in each animal
species.
The clinical signs do not vary greatly with serotypes of Leptospira
except that infection with Leptospira interrogans serovar ictero
haemorrhagiae usually causes a severe septicemia.
We will see the clinical signs in each species of animals.
1. Cattle
Leptospirosis in cattle is usually associated with two serovars
I. L.interrogans serovar pomona or
II. L. interrogans serovar hardjo
I. Leptospirosis in cattle associated with L.interrogans serovar
pomona 686
 Lepto. cont’d---
Leptospirosis which is caused by this serovar can be
Acute
Subacute
Chronic
1. Acute leptospirosis in cattle
It is usually associated with L. interrogans serovar pomona.
Calves upto 1 month old are most susceptible to the acute
leptospirosis.
The disease is manifested by
Septicemia with high fever (40.5 - 41.5 °C)
Anorexia and petechiation of mucosae,
Depression and acute hemolytic anemia with haemoglobinuria
Jaundice and pallor of the mucosae
 Dyspnea is also prominent
 The case-fatality rate is high and if recovery occurs convalescence
is prolonged.
In adult cattle, abortion due to the systemic reaction may occur at
687
 Lepto. cont’d---
at the acute stage of the disease.
Milk production is markedly decreased and the secretion is red
colored or contains blood clots and the udder is limp and soft.
Mastitis as part of leptospirosis has often been described in cattle
and a high somatic cell count in grossly abnormal milk suggests
mastitis.
But these changes are due to a general vascular lesion rather than
local injury to mammary tissue.
Severe lameness due to synovitis is recorded in some animals and a
necrotic dermatitis probably due to photosensitization, in others.
2. Subacute leptospirosis in cattle
It is usually associated with L. interrogans serovar pomona.
The subacute form of leptospirosis differs from the acute form only
in degree.
Similar clinical findings are observed in a number of affected
animals .
But not all of the findings are present in the same animal. 688
 Lepto. cont’d---
The fever is milder (39 - 40 0C) and depression, anorexia, dyspnea
and haemoglobinuria are common.
But jaundice may or may not be present.
Abortion usually occurs 3 - 4 weeks later.
One of the characteristic findings is the marked drop in milk
production and the appearance of blood-stained or yellow-orange,
thick milk in all four quarters without apparent physical change in
the udder.
3. Chronic leptospirosis in cattle
It is associated with L. interrogans serovar pomona.
The clinical findings in the chronic form of leptospirosis are mild
and may be restricted to abortion.
Severe 'storms' of abortions occur most commonly in groups of
cattle which are at the same stage of pregnancy when they are
exposed to infection.
The abortions usually occur during the last trimester of pregnancy.
In rare case many animals in the group develop positive 689
 Lepto. cont’d---
agglutination titers without clinical illness.
There are occasional reports of leptospiral meningitis in cattle.
The clinical signs are
 In coordination
Excessive salivation
Conjunctivitis
Muscular rigidity are the common signs of meningitis caused by L.
interrogans serovar pomona.
II. Listeriosis in cattle associated with L.interrogans serovar
hardjo
The main clinical signs of listeriosis in cattle those are caused by
L.interrogans serovar hardjo are
Abortion
Infertility
Milk drop syndrome
Abortion may occur several weeks after the initial infection and
may also occur as the only evidence of the disease. 690
 Lepto. cont’d---
In some areas or circumstances it is the principal clinical
manifestation of leptospirosis due to Leptospira interrogans serovar
hardjo.
Infertility and milk drop syndrome occurs only in pregnant or
lactating cows.
Because the organism is restricted to proliferation in the pregnant
uterus and the lactating mammary gland.
There is a sudden onset of fever, anorexia, immobility and agalactia.
The milk is yellow to orange and may contain clots.
The udder is flabby, there is no heat or pain, and all four quarters
are equally affected.
The sudden drop in milk production may affect up to 50% of cows
at one time and cause a precipitate fall in the herd's milk yield.
The decline may last for up to 8 weeks but in individual cows milk
production will return to normal within 10 - 14 days.
The milk may have a high leukocyte count.
In some cases, there is no evidence of mastitis, no change in the 691
 Lepto. cont’d---
consistency of the milk and no changes in the udders of affected
cows.
But leptospiruria may be present in up to 30% of affected cows.
The herd fertility status can be known by considering
First service conception rate
The number of services per conception for cows conceiving
Calving to conception interval
The culling rate
They usually reveal a low reproductive performance especially
during the year of the diagnosis.
Exposure of non vaccinated dairy cows to L . interrogans serovar
hardjo can be associated with a subsequent reduction in fertility, as
indicated by
Greater time from calving to conception and/ or
 Higher number of breedings per conception
2. Leptospirosis in pigs
Leptospirosis in pigs is caused commonly by 692
 Lepto. cont’d---
Leptospira interrogans serovar pomona
Leptospira interrogans serovar tarrassovi
The infection is characterized by
 Abortion and
A high incidence of stillbirths
However, failure to conceive is not usually observed in leptospirosis
of pigs caused by the above serovars.
But failure to conceive in pigs has been reported in infections with
Leptospira interrogans serovars canicola.
An abortion' storm‘ may occur when the disease first appears in a
herd.
Most abortions occur 2 - 4 weeks before term.
Piglets produced at term may be dead or weak and die soon after
birth.
But abortions diminish in subsequent pregnancy as the herd
develops immunity.
L. interrogans serovar hardjo may cause sporadic reproductive 693
 Lepto. cont’d---
disease in pigs.
However, L. interrogans serovar muenchen and bratislava are
occasionally isolated during investigations of porcine abortion and
stillbirths.
Listeriosis in piglets caused by L. interrogans serovar
icterohaemorrhagiae causes septicemic leptospirosis with a high
mortality rate.
3. Leptospirosis in sheep and goats
Leptospirosis in sheep and goats is caused by
Leptospira interrogans serovar pomona
Leptospira interrogans serovar hardjo
It may have
Acute form
Chronic form
In acute form, most affected animals are found dead apparently
from septicemia.
Affected animals are 694
 Lepto. cont’d---
Febrile
Dyspneic
Snuffle and hang their heads down
Some have haemoglobinuria, pallor of mucosae and jaundice, and
die within 12 h.
Abortion seems to be almost entirely a manifestation of the acute
form of listeriosis when the infection is caused by L. interrogans
serovar pomona.
Lambs, especially those in poor condition are most susceptible.
The chronic form may occur and is manifested by loss of bodily
condition.
When infection is caused by L. interrogans serovar hardjo, abortion
has been recorded as the only clinical sign.
Oligolactia and agalactia, similar to the bovine milk drop syndrome
have been observed in lactating ewes by L. interrogans serovar
hardjo.
695
 Lepto. cont’d---
4. Leptospirosis in horses
Leptospirosis in horses is characterized by
Abortion
Periodic ophthalmia
Leptospira interrogans serovar pomona is the major cause of
abortions and stillbirths in equines.
The gestational ages when abortion takes place range from 140 -
250 days .
Periodic ophthalmia also called recurrent uveitis, moon blindness
and recurrent iridocyclitis is often associated with infection with L.
interrogans serovar pomona.
Clinically there are recurrent episodes of ocular disease including
Photophobia
Lacrimation
 Conjunctivitis
Keratitis
 A Apericorneal corona of blood vessels 696
 Lepto. cont’d---
Hypopyon and iridocyclitis.
Recurrent attacks usually terminate in blindness in both eyes.
There is a strong relationship between uveitis and leptospiral
seropositivity in horses.
Seropositive horses with uveitis are at increased risk of losing
vision compared with seronegative horses with uveitis.
Infection with L. interrogans serovar pomona in foals has been
observed in association with Rhodococcus equi to cause a very
heavy mortality.
The foals died of a combination of interstitial nephritis and uremia
and pulmonary abscessation and chronic enteritis.
Nonulcerative keratouveitis associated with leptospiral infection
has been described in horses with clinical signs of
Photophobia
Epiphora and blepharospasm
Hyperemia of the bulbar conjunctiva
697
 Lepto. cont’d---
Epiphora and blepharospasm
Hyperemia of the bulbar conjunctiva
Edema of the paralimbal cornea
Papillary block and iris bombe are also present
Necropsy findings
Acute bovine leptospirosis is characterized by
Anemia
Jaundice
 Haemoglobinuria
 Subserosal hemorrhages
There may be ulcers and haemorrhages in the abomasal mucosa.
Pulmonary edema and emphysema are also common in this species.
Histologically, there is focal or diffuse interstitial nephritis and
centrilobular hepatic necrosis.
Aborted bovine fetuses are usually autolyzed to the point where no
lesions or bacteria can be demonstrated.
Even in a fresh fetus the positive identification of leptospira in 698
 Lepto. cont’d---
lesions is not an easy task.
Gross placental lesions in cases of equine abortion and stillbirth
associated with leptospirosis include
Nodular cystic allantoic masses
Diffuse edema
 Areas of necrosis with a mucoid exudate on the chorionic surface.
 The liver is enlarged, mottled and pale-red to yellow.
The kidneys are swollen and edematous with pale, radiating streaks
in both cortex and medulla.
Aborted piglets are usually severely autolytic with blood- stained
fluid in the subcutis and filling the body cavities.
Diagnosis
Diagnosis of leptospirosis is done based on
Epidemiology of the disease
Clinical signs of the disease
Necropsy findings
Laboratory method 699
 Lepto. cont’d---
Laboratory procedures used in the diagnosis include
1. Bacteriology
2. Serology
3. Animal inoculation
4. Molecular techniques
1. Bacteriology
Bacteriology includes isolation and identification of leptospires in
appropriate specimens like blood or body fluids.
Samples for confirmation of diagnosis include
Whole blood( 0.5 ml) in leptospiraemic stage for bacterial isolation
Midstream urine
A small plug of kidney taken from inside the organ with sterile glass
Pasteur pipette for dark – field microscopy after maceration with a
little distilled water
Kidney and other body tissues(liver).
 Placenta, foetal abomasal content, foetal kidney, cotyledon and
uterine discharge 700
 Lepto. cont’d---
During septicemic stage leptospirae are present only in the blood
and there may be
Laboratory evidence of acute hemolytic anemia and increased
erythrocyte fragility and often haemoglobinuria.
However, the only positive diagnostic measure at this stage of the
disease is culture of the blood.
If abortion occurs, the kidney, lung and pleural fluid of the aborted
fetuses should be examined for the presence of the organism.
NB. The zoonotic potential of this organism should be noted when
handling carcasses and submitting specimens.
Demonstration or culture of organism
There are many methods to detect leptospires directly from
specimens
These are
1.Dark field microscopy
2. Silver impregnation technique
3. Serology(FAT, immunoperoxidase staining technique etc) 701
 Lepto. cont’d---
 4. Molecular techniques
Procedure to prepare the specimens for dark field microscopy and
silver impregnation techniques
1. Dark field microscopy
A. If the specimen is urine
Centrifuge the urine at 9750RCF( Relative Centrifugal Force) for 10
minutes to concentrate the leptospires.
Then use the sediment for dark field microscopy
B. If the specimen is blood
Unclotted blood with anticoagulant is centrifuged at low speed to
sediment red blood cells.
Take the plasma and centrifuge it again at 9750 RCF for 10 minutes.
Take the supernatant for dark field microscopy
C. If the specimen is tissue
Tissue specimens are grind in tissue grinder
Then prepare a 10 % tissue suspension in 1% Bovine Serum
Albumin (BSA). 702
 Lepto. cont’d---
Take a drop of tissue suspension for dark field microscopy
Procedure
Put the drop of the above preparations on microscope slide .
Cover with cover slip.
Examine under dark – field microscopy.
In all cases look the prepared smear under dark – field microscope.
Under dark – field microscopy use low power dry objective if large
numbers of leptospires are present.
But if the number of the bacteria are small, use high power dry
objective or oil – immersion objectives.
As the result, the bacteria appear as silver, beaded ( due to the
helical coils), slender rods with distinct hooks at each ends.
In dark field microscopy the L. interrogans are obviously motile if
they are viable.
They have flexuous movements and spinning around the long axis.
2. Silver impregnation
It can be used on tissue sections of kidney, liver or other tissues. 703
 Lepto. cont’d---
Leptospira under dark – field microscopy

704
 Lepto. cont’d---
Leptospira interrogans serovar hardjo under silver impregnated
stain

705
 Lepto. cont’d---
2. Serology
The most common serological methods to diagnose leptospirosis is
FAT
Immunoperoxidase staining techniques
Macro and micro agglutination
ELISA
CFT
Serological testing at the time of abortion is often unreliable.
Because the acute titers have already peaked and are declining.
In the stage immediately after the subsidence of the fever, antibodies
begin to develop and the leptospirae disappear from the blood and
appear in the urine.
The diagnosis of leptospirosis is much easier on a herd basis than in
a single animal.
Because in an infected herd, some animals are certain to have high
titers and the chances of demonstrating or isolating the organism in
urine or milk are increased with samples being taken from many 706
 Lepto. cont’d---
L. interrogans serovar hardjo in a bovine urinary
deposit by direct FAT

707
 Lepto. cont’d---
Leptospiral antigen in the luminal border of tubular epithelium in
the Kidney of pig stained by immunoperoxidase

708
 Lepto. cont’d---
Leptospira spp. in the kidney of pig stained by immunoperoxidase

709
 Lepto. cont’d---
animals.
On the other hand, in a single animal, depending on when the
infection occurred, the titer may have declined to a low level and be
difficult to interpret.
Macroscopic and microscopic agglutination tests and to lesser extent
CFT and ELISA techniques are used for the detection of leptospiral
antibodies in serum.
The macroscopic agglutination test is a screening uses dead antigen
and has low specificity.
The Microscopic Agglutination Test (MAT) is the most commonly
used serological test for the diagnosis of leptospirosis.
The leptospiral microscopic agglutination test uses live leptospires
as an antigen and is highly sensitive and serum – specific.
In animals which survive infection, acute leptospirosis can readily
be diagnosed on the basis of demonstrating a rising antibody titer in
acute and convalescent sera.
Paired sera are normally considered necessary so that a rise in titer
710
 Lepto. cont’d---
can be detected, in cases of bovine abortion or stillbirth, infection.
A MAT titer of >100 is considered positive.
The ELISA test is much more accurate than the others and has many
advantages.
The most common ELISA tests are
Direct ELISA
In direct ELISA
NB. Measurement of aqueous humor antibody titers against
leptospires in horses offers a more accurate means of establishing a
diagnosis of leptospiral-associated uveitis than serology alone.
Antibody can be detected in cervico-vaginal by ELISA as early as
2 weeks after natural or experimental infection and may reach high
levels after 8 weeks.
4. Molecular technology
DNA probes, PCR and nucleic acid hybridization for detection of
leptospires in semen and in aqueous humor of horses with uveitis
are commonly used. 711
 Lepto. cont’d---
Animal inoculation
Hamsters and guinea pigs can be inoculated intraperitoneally.
Use 0.5 – 1ml of neutralized urine or unclotted or 10 % tissue
suspension in EMJH or 1% BSA.
Then take the cardiac blood from these laboratory animals
aseptically when their body temperature rise.
Or take the cardiac blood of these animals at 5, 8, 10 and 14 days
post inoculation if there is no temperature reaction.
 Inoculate into the medium with 2 – 3 drops of freshly collected
blood from these laboratory animals
 Then incubate the inoculated medium and follow the media as
above.
Lastly take blood from these animals if they are alive after 21 days.
Extract serum and use it for serology.
Differential diagnosis
Acute leptospirosis in cattle must be differentiated from those
diseases causing haemolytic anemia with or without 712
Summary of laboratory procedures for diagnosis of leptospira

713
 Lepto. cont’d---
haemoglobinuria
They include
Babesiosis
Anaplasmosis
 Rape and kale poisoning
Post parturient haemoglobinuria
Bacillary haemoglobinuria
Chronic leptospirosis causing abortion must be differentiated from
all other causes of abortion in cattle including
Infectious bovine rhinotrachitis(IBRT)
 Protozoal abortion caused by Sarcocytis spp. and Toxoplasma gondi
 Brucellosis
Bovine Virus Diarrhea(BVD)
Pine needle abortion
 Mycotic placentitis
 Campylobacteriosis
Abortion caused by Actinomyces pyogenes 714
 Lepto. cont’d---
Abortion caused by Ureaplasma
Milk drop syndrome caused by Leptospira spp. is characterized by a
sudden drop in milk yield in upto 30 - 50% of the cows within
several days.
It must be differentiated from other causes of declining in milk
production of the herd.
These are
Change of feed
Change of management
 Other sever infectious diseases accompanied with fever.
In pigs abortion due to leptospirosis occurs in the last trimester.
The common manifestation of leptospirosis in pigs and must be
differentiated from
All other causes of abortion, mummification and stillbirths in swine.
Other common causes of abortion in swine are
Parvovirus
715
Porcine reproductive respiratory syndrome.
 Lepto. cont’d---
Brucellosis
 Pseudo rabies
 Stillbirth, mummification and Early Embryonic Death(EED)
In sheep and goats leptospirosis must be differentiated from
Chronic copper poisoning and poisoning caused by rape in sheep.
The clinical picture may present a similar to that in leptospirosis.
But there will be no febrile reaction.
Anaplasmosis associated with Anaplasma ovis may b e
accompanied by fever and haemoglobinuria.
But it is more commonly a chronic and emaciating disease and it can
be demonstrated by blood smear in Giemsa staining from peripheral
ear vein.
In horses leptospirosis must be differentiated from
Abortion, stillbirths, perinatal deaths of foals caused by
Streptococcus zooepidermicus, Salmonella abortus equi, Escherichia
coli and Actinobacillus equi.
Infectious equine anemia
716
Chronic periodic ophthalmia
 Lepto. cont’d---
Differential diagnosis of cattle characterized by acute haemolytic anaemia with
or with out haemoglobinuria
Disease Epidemiology Clinical findings Laboratory findings
Leptospirosis All age, cattle on Acute fever, red-colored
pasture milk.
Haemoglobinuria •Leptospira titers
abortion.
May die in 24-48 hours
Postpaturient High producing Acute , No change in milk,
haemoglobinuria lactating cows 4 No fever.
– 6 weeks Die with in 12 – 48 hours •Hypophosphatemia
postpartum Marked haemoglobinuria
Bacillary Usually mature Acute fever, abdominal
haemoglobinuria on summer in pain •Leukopenia or
enzootic area May die in 2 – 4 days •leukocytosis
Haemoglobinuria
Babesiosis Enzootic areas, Acute fever, jaundice, •Blood smear
tick-borne, abortion •CFT
Affect young course of 2-3 weeks.
animals Marked 717
haemoglobinuria
 Table cont’d---
Differential diagnosis of cattle characterized by acute haemolytic anaemia with or with
out haemoglobinuria
Anaplasmosis Yearling and mature No Anaplasmas on
cattle, common in haemoglobinuria, blood smear and
summer, insect-borne. jaundice common, CFT
common in feedlots fever
Chronic copper Follows long term oral Severe jaundice. No Toxic levels of
poising administration medicines fever. copper in blood,
and food contain copper Haemoglobinuria liver and faces

Rape and kale All ages of cattle Peracute hemolytic Acute hemolytic
poisoning rape crop grown for anemia, may die in anemia
F odder in fall few hours anemia
Drug-induced Some drug preparations Mild Nil
haemolytic anaemia when given intravenously haemoglobinuria
No haemolytic
anaemia
Blood transfusion Using blood from same Sudden onset, Nil
reaction donor more than 1 week dyspnea, hiccoughs,
after initial transfusion trembling responds
718
to adrenaline
 Table cont’d---
NB. The common causes of haematuria in cattle are pyelonephritis
and cystitis due to Corynebacterium renale, non-specific cystitis and
enzootic haematuria.
Myoglobinuria occurs occasionally in young cattle affected with
enzootic-nutritional muscular dystrophy and may be confused with
haemoglobinuria.
Treatment
The aim of treatment of leptospirosis are
Controlling the infection before irreparable damage to the liver and
kidneys occurs
 Controlling the leptospiruria of 'carrier' animals and render them
safe to remain in the to remain in the group.
The methods to treat animals from leptospirosis are
1. Parenteral administration of antimicrobials like
Dihydrostreptomycin (12 mg/kg BW IM twice daily for 3 days) is
effective in the treatment of the systemic infection.
Other antimicrobial drugs that are effective to treat leptospirosis719
are
 Table cont’d---
Oxytetracycline
Tilmicosin
 Ceftiofur and amoxicillin are also effective for resolving
leptospirosis in cattle.
For the elimination of leptospiruria in cattle and pigs a single dose
of
Dihydrostreptomycin (25 mg/kg BW IM) is recommended.
Long-acting oxytetracycline at 20 mg/kg BW is an alternative
recommendation.
For outbreaks of leptospiral abortion in horses, treatment of pregnant
mares with dihydrostreptomycin (50 mg/kg BW)IM daily for 3 - 5
days can minimize further abortions.
For equine periodic ophthalmia caused by leptospirosis
A course of a suitable antibiotic systemically and the administration
of a corticosteroid either parenterally in an acute episode or
Subconjunctivally suitable antibiotics with corticosteroids in a
chronic case is most likely to be satisfactory. 720
 Lepto. Cont’d---
Nonulcerative keratouveitis require long-term and intensive
medication, and recur with tapering of treatment.
 Topical and subconjunctivally corticosteroids are recommended in
controlling nonulcerative keratouveitis.
Atropine eye ointment is also usually applied three times daily to
maintain dilatation of the pupil.
2. Administration of antimicrobials in feed
 In groups of pigs, feeding of oxytetracycline (800 g/tone of feed
for 8-11 days) is claimed to eliminate carriers.
 Antimicrobial feeding should begin 1 month before farrowing to
avoid the occurrence of abortion.
3. Supportive treatment with blood transfusions.
Blood transfusions in dose of 5-10 liter/ 450 kg BW are indicated as
supportive treatment for the hemolytic anemia in acute leptospirosis
in cattle.
 Systemic administration of Vitamin B6, B12 and preparates those
have iron are essential. 721
 Lepto. cont’d---
Prevention and control measures
Prevention and control measures of leptospirosis comprises
1. Biosecurity and biocontainment measure
You know sources of leptospiral infection include
Clinically affected animals
Carrier animals
Wildlife, dogs, cats and rodents
 Environmental sources such as water supplies.
The probability of infection of animals by leptospira serovars is
increased by four factors:
1. Purchase of infected cattle
2. Co -grazing or common grazing with infected cattle or sheep
3. Use of natural service with an infected bull
4. Use of infected semen in artificial insemination
5. Access of cattle to contaminated water sources such as streams,
rivers, flood or drainage water
6. Contact of animals with wild animals and rodents 722
 Lepto. cont’d---
So that controlling the above methods of introduction of leptospiral
infections reduces the occurs of the disease in your farm.
For example: -
In artificial insemination centers, bulls destined as a source semen
must be free of antibody to different serovars of Leptospira
interrogans serovar hardjo, grippotyphosa, canicola, pomona and
icterohaemorrhagiae at a final serum dilution of 1:100 in the MAT.
To know whether the bulls are free from leptospiral infection or not
Culture of leptospires from semen of bulls is possible.
But it is difficult, costy and time consuming.
However, a PCR assay has been developed to detect pathogenic
leptospires in the semen and urine of infected bulls.
2. On an individual farm, leptospirosis can be eradicated or
controlled by vaccination
Vaccination against leptospirosis in cattle and swine is in general
use and an effective method for control of the disease
This suggests that vaccine manufactures should consider using the 723
 Lepto. cont’d---
genotypes which are most prevalent in cattle and pigs.
Vaccination of cows in late pregnancy gives effective immunity to
their calves.

724
Here we will focus on
I. Introduction to mastitis
II. Diagnostic methods of mastitis
III. Mastitis in cattle
IV. Mastitis in domestic animals other than cattle
V. Complication of mastitis

725
 Lepto. cont’d---
I. Introduction to mastitis
Mastitis is one of the first observed diseases of farm animals.
It was observed when cattle were domesticated over 5000 years ago.
Mastitis is inflammation of the parenchyma of the mammary gland
regardless of the cause.
Mastitis can have an infectious or non-infectious etiology.
Mastitis is also defined as inflammation of mammary gland
parenchyma which is caused by non - infectious agents and / or
microorganisms usually bacteria that invade the udder, multiply and
produce toxins which are harmful to the mammary gland.
Mastitis affects the quality and quantity of milk.

726
 Mastitis Cont’d---
Mastitis is therefore characterized by
Range of physical, chemical and bacteriological changes in milk
 Pathological changes in the glandular tissue
The most important physical and chemical changes in the milk
include
Discoloration
The presence of clots and
The presence of large numbers of leukocytes (neutrophils)
That is why mastitis affects the quality and quantity of the milk.
Depending on the clinical manifestations on the udder mastitis can
be
1. Clinical mastitis
2. Subclinical mastitis
In many clinical cases of mastitis, the following clinical features are
manifested in mammary gland.
These are
Swelling 727
 Mastitis Cont’d---
Heat
Pain
Edema
However, large proportion of mastitic glands are not readily
detectable by manual palpation nor by visual examination of the milk
using a strip cup.
This type of mastitis represent subclinical mastitis.
It seems practicable and reasonable to define mastitis as a disease
characterized by the presence of a significantly increased
SCC(Somatic Cell Count) in milk from affected glands.
The increased SCC is, in almost all cases, due to an increased
leucocytes mainly neutrophil concentration.
However, the exact clinical and laboratory changes that occur in the
udder as a result of infection can also be caused by other factors in
the absence of infection(Physical and chemical factors).
It is known that mastitis affects all species of female domestic
animals including bovine, ovine, caprine, swine and equine. 728
 Mastitis Cont’d---
However, we will focus entirely with bovine mastitis because of its
economic importance.
But small sections on ovine, caprine, porcine and equine mastitis are
included in the chapter.
Mastitis is a multi - etiological complex disease.
The etiological factors can be
1. Physical agents
2. Chemical agents
3. Biological agents
1. Mastitis caused by physical agents
Mastitis is caused by different physical agents like trauma due to
coldness, mechanically and incorrect exploitation of milking
machine etc
All mastitis that are caused by different physical agents can be
grouped in traumatic mastitis.
As injuries to the teats or udder that penetrate to the teat cistern or
milk ducts. 729
 Mastitis Cont’d---
It can also involve to the external sphincter, are commonly followed
by mastitis.
Any of the organisms that cause mastitis may invade the udder after
such injury.
In such cases mixed infections are usual.
So that all injuries to the teat or udder including surgical
interference should be treated prophylactically with broad-spectrum
antibiotics.
2.Mastitis caused by chemical agents
Mastitis which is caused by different chemicals that act on udder
called mastitis caused by chemicals.
Mastitis caused by physical and chemical agents are called non –
infectious mastitis.
Mastitis which are caused by physical and chemical agents can
expose the animal to infectious mastitis.
3. Mastitis caused by biological agents
Mastitis caused by different biological agents like bacteria, virus,730
 Mastitis Cont’d---
fungi and rarely by some algae is called biological mastitis.
It is one of the most common mastitis in most dairy farms of the
world,
Based on their epidemiology and pathophysiology, these
pathogens have been further classified as causes of
1. Contagious mastitis
2. Teat skin opportunistic mastitis
3. Environmental mastitis
4. Mastitis caused by less common pathogens
5. Mastitis caused by different specific diseases called
miscellaneous infectious conditions

731
 Mastitis Cont’d---
II. Diagnostic methods of mastitis
Diagnostic methods of bovine mastitis depend on the type of
mastitis(clinical or subclinical).
Clinical mastitis can be diagnosed based on
Routine physical examination of the udder, supramammary lymph
nodes and the milk.
The health condition of the cow if there is systemic infection
However, laboratory culturing of milk samples for isolation and
identification bacteria is a gold standard method to diagnose
clinical mastitis.
This method helps us
 To select the most effective antimicrobial drugs for treatment by
doing antimicrobial sensitivity test.
To design the appropriate control measures
We know that subclinical mastitis can not be detected by using
routine physical examination as clinical mastitis.
In other words, it is possible to say 732
 Mastitis Cont’d---
The affected quarters and supramammary lymph nodes do not show
the cardinal signs of inflammation
 The secretion from a quarter with subclinical mastitis appears
drinkable.
But subclinical mastitis has the greatest economic influence on the
dairy farms.
So that it is advantageous to diagnose subclinical mastitis on time,
on a cow and quarter level for early detection of subclinical mastitis.
We know culturing of the milk samples for isolation and
identification of the bacteria is a gold standard method to
diagnose subclinical mastitis.
However, this method is expensive and impractical for routine use.
Nowadays, there are two indirect methods to diagnose subclinical
mastitis.
The two methods are mainly based on determination of Somatic
Cells in the milk called Somatic Cell Counts(SCC).
SCC is quantified as cells per ml of milk. 733
 Mastitis Cont’d---
These cells can be counted by
 1. Direct cell counting methods
 2. Indirect test method
1.Direct cell counting methods
Many direct cell counting methods have been developed.
However, the most common methods are
Electronic somatic cell counts
Direct microscopic count on stained milk samples
Automated device for rapid determination of SCC in milk samples
Electronic somatic cell count uses equipments such as a coulter
counter.
The device counts the total cell counts as both exfoliated epithelial
cells and leukocytes.
In the case of direct microscopic count on stained milk samples,
only leukocytes are counted directly.
For this, a known volume of milk(0.01 ml) is spread over 1 cm2 on a
734
microscopic slide
 Mastitis Cont’d---
Stain by methylene blue based stain.
Since the microscope is calibrated and from an average number of
leucocytes per field( counting 50 fields).
From this number of leukocytes/ ml of milk can be calculated.
NB. If comparatively large number of pathogenic bacteria are
present in the milk sample, these may also be seen in the stained
smear.
The most commonly used device which is automated device for
rapid determination of SCC in milk samples is the Fossomatic
milk cell counter.
This instrument stains cells with a fluorescent dye and then counts
the number of fluorescing particles.

735
Direct microscopic count on stained milk samples

736
Pathogenic bacteria on stained smear

737
Direct SCC in the milk using fluorescence microscope

738
Fossomatic milk cell counter

739
 Mastitis Cont’d---
2. Indirect test method
Nowadays, the direct methods of diagnoses of subclinical mastitis
are not widely applicable in the field.
Because they need expensive equipments, experienced personnel
and laboratory reagents.
So much attention has been given to the development of indirect
test methods to predict the presence of an intramammary infection.
Currently available indirect tests of subclinical mastitis are
1. California Mastitis Test (CMT)
2. The NAGase (N- acetyl-β-D-glucosaminidase) test
3 . Electrical conductivity of milk test
4. Catalase test
Of these indirect tests, only the CMT and electrical conductivity are
commonly used methods.
All of them are screening tests.
Because they show only the presence of inflammation in the
udder milk from quarters or cows. 740
 Mastitis Cont’d---
That is why milk samples those are positive for subclinical mastitis
in screening test must be submitted to bacteriological laboratory to
isolate and identify the causative agent.
1. California Mastitis Test (CMT)
CMT is also called rapid mastitis test, Schalm test or Mastitis-N-K
test.
CMT is simple a modification of the Whiteside test which
described a reaction between sodium hydroxide and milk that
resulted in the thickening of mastitic milk.
The utility of Whiteside test as a field test was limited by the fact
that
the reaction was sometimes difficult to observe and
It would eventually occur even in normal milk( low specificity of
the test)
A refined version of the test, which enhanced its sensitivity, and
eliminated the confounding effect of milk fat, uses an anionic
surfactant with bromocresol purple indicator is called CMT. 741
 Mastitis Cont’d---
It forms a gel with the DNA of somatic cells in the milk.
It is one of a reliable screening test used to diagnose subclinical
mastitis.
This test can be used in the field or in the laboratory.
The CMT reagent contains
 A detergent(surfactants) that reacts with DNA of cell nuclei
 PH indicator (bromocresol purple) that changes color when the
milk PH is increased above its normal value of approximately 6.6
(mastitis increases PH to 6.8 or above).
CMT is based on the quantity of DNA in the milk and hence the
number of leukocytes and other cells present.
It operates by disrupting the cell membrane of any cells present in
It operates by disrupting the cell membrane of any cells present in
the milk sample.
As the result the DNA in the cells comes out and react with the test
reagent, forming a gel.
The procedure of CMT is as follows: 742
 Mastitis Cont’d---
Take a squirt of milk from each quarter of the udder
Place the milk in each of four shallow cups in the CMT paddle
Add equal amount of 3 % commercial CMT reagent to each cups
Mix them by gentle circular motion in a horizontal level.
A positive gelling reaction occurs within 15 seconds but maximum
gel formation actually occurs from 1 - 2.5 minute
Depending on the thickness of the gel which is formed due to the
reaction of CMT reagent and milk sample, the results are scored
as
 0 (negative)
 T(trace)
 1 (weak positive)
 2 (distinct positive)
 3 (strong positive).
Milk samples with test result of CMT 1 to 3 were classified as
evidence of subclinical mastitis.
743
 Mastitis Cont’d---
NB. Cows in the first week after calving or in the last stages of
lactation may give a strong positive reaction(False positive) test
result.

744
 Mastitis Cont’d---
Interpretation for the California Mastitis Test (CMT) result
Score Description Interpretation
N(Negative) No change Healthy quarter
T (Trace) Slime formed which disappeared with Subclinical mastitis
continuous movement of paddle
1 (Weak) Distinct slime, but no gel formation Subclinical mastitis
which do not disappear with
continuous movement of paddle.
2(Distinct Viscous with gel formation, which Serious-mastitis
positive) adhered to the margin. infection
3(Strong positive) The gel formation with convex Serious-mastitis
projection, the gel did not dislodge infection
after swirling movement of the paddle
745
Californian Mastitis Test(CMT) and test results

Interpretation
Normal 0(Top left)
Positive 1(Top right)
Positive 2( Bottom right)
Positive 3( Bottom left) 746
Procedure of CMT and test result

747
 Mastitis Cont’d---
Correlation of gel formation with somatic cell count in CMT
CMT score Somatic cell range per ml of milk(SCC/ ml ) Gelling
None 0 - 200, 000 None
Trace 200, 000 - 400, 000 Very mild
1 400, 000 - 1,200000 Mild
2 1, 200,000 – 500, 000 Moderate
3 Over 5000,000 Heavy almost
solidifies

2. The NAGase (N- acetyl-β-D-glucosaminidase) test

The NAGase test is based on the measurement of a cell-associated
enzyme (N- acetyl-β-D-glucosaminidase) in the milk.
So that high enzyme activity indicating a high cell count.
NAGase is an intracellular lysosomal enzyme derived primarily
from neutrophils but also from damaged epithelial cells. 748
 Mastitis Cont’d---
NAGase test must be done on fresh milk and read on the same day.
However, because most of the NAGase activity is intracellular,
samples should be frozen and thawed before analysis to induce
maximal NAGase activity.
3 . Electrical conductivity of milk test
It is based on the increase in concentration of sodium and chloride
ions in mastitic milk.
Which consequently increase in electrical conductivity.
The electrolyte changes in milk are the first to occur in mastitis and
the test has attractions for this reason.
4. Catalase test
The test used to know the presence of an enzyme catalase that
breaks hydrogen peroxide in to water and oxygen.
Since mastitic milk has large amount of somatic cells, large amount
of oxygen gas will be produced which can be detected by sound
which is produced.
749
 Mastitis Cont’d---
CMT is the most common screening test to detect subclinical
mastitis.
Subclinical mastitis can be diagnosed using CMT
1.At the herd level
2. At individual animal level
1.Detection of mastitis at the herd level
The prevalence of subclinical mastitis or intramammary infection is
monitored by
Determining the bulk tank milk SCC and
Isolation and identification of the most likely mastitis pathogens in
the laboratory from bulk tank milk
These two methods are recommended to diagnose the presence and
prevalence of mastitis pathogens on a herd basis.
The sample for analysis is obtained by agitating the milk for 5 -10
minutes and collecting a sample from the top of the bulk tank milk
using a clean dipper.
Milk processing plants in most developed countries use routinely750.
 Mastitis Cont’d---
automatic electronic somatic cell counters .
The device provides a monthly report of the bulk tank milk SCC.
The test requires only that the sample for examination be taken
randomly and not frozen.
The milk sample must be fresh and to be examined quickly with in
2 hours.
Other wise it must be preserved with formalin to prevent cell losses
during storage.
The bulk tank milk SCC is extremely useful in creating awareness
of the existence of a mastitis problem.
So that when the SCC of bulk tank milk exceeds permissible limits
further investigation of the herd is indicated.
But you must remember SCC in the bulk tank milk may increase
not only during inflammation but also in some cases like
Cows in early and late lactation
The presence of Corynebacterium bovis in teat canal (with out
inflammation of mammary glands). 751
 Mastitis Cont’d---
It is not possible to use the bulk tank milk SCC to determine the
number of cows in a herd affected by subclinical mastitis.
But it is possible to estimate fairly accurately infected quarters.
In general, the increasement of the bulk tank milk SCC shows
The increasement of the prevalence of udder infection and
The loss of production in the farm
That is why herds with a high bulk tank milk SCC have
 A significant low production levels and
 They are less likely to use a post milking teat dip or to have a
regular program of milking machine maintenance and automatic
cluster removal.

752
 Mastitis Cont’d---
Relation of SCC in bulk milk with infected quarters and loss of
production
Bulk tank milk SCC/ ml Infected quarters in the Production loss in %
herd
200000 6 0
5000000 16 6
1000000 32 18
1500000 48 29

NB.A bulk tank milk SCC of more than 300000 cells/ml is

considered to indicate a level of mastitis in the herd that warrants
examination of individual cows.

753
 Mastitis Cont’d---
II. Mastitis in cattle
Bovine mastitis is defined as inflammation of mammary gland
parenchyma of cattle which is caused by none - infectious
Agents(physically And chemically) and / or microorganisms usually
bacteria, Fungi and rarely virus that invade the udder, multiply and
produce toxins which are harmful to the mammary gland.
A total of about 140 microbial species, subspecies and serovars
have been isolated and identified from the bovine mammary gland.
Etiology
Bovine mastitis is caused by many different physical, chemical
and biological agents.
However the most common causes of bovine mastitis are infectious
agents.
All of infectious agents those causing bovine mastitis are divided
into
1. Mastitis of cattle associated with common contagious pathogens
2. Mastitis of cattle associated with common environmental 754
 Mastitis Cont’d---
pathogens
 3.Mastitis of cattle associated with common opportunistic
pathogens
4. Mastitis of cattle associated with less common pathogens
5. Miscellaneous causes of bovine mastitis
1. Mastitis of cattle associated with common contagious pathogens
Here we will see
1.1. Mastitis of cattle caused by Staphylococcus aureus
1.2.Mastitis of cattle caused by Streptococcus agalactiae
1.3.Mastitis of cattle caused by Mycoplasma bovis
1.4. Mastitis of cattle caused by Corynebacterium bovis
1.1. Mastitis of cattle caused by Staphylococcus aureus
As the other contagious mastitis, it is mastitis which can spread
from infected quarters to other quarters and from infected cows to
healthy herds.
Etiology
Coagulase-positive S. aureus is a major pathogen of the bovine 755
 Mastitis Cont’d---
mammary gland.
It is the common cause of contagious mastitis in cattle.
S. aureus also causes mastitis in sheep and goats.
Epidemiology
Occurrence and prevalence of infection
S. aureus was one of the most common causes of bovine mastitis
in dairy cattle.
The prevalence of infection and the occurrence of clinical mastitis
due to S. aureus has decreased in herds using effective mastitis
control measures.
However, surveys indicate that 50 - 100% of herds can be
infected in dairy farms with out effective mastitis control
measures.
The majority of intramammary infections due to S. aureus are
subclinical.
That is why with an effective mastitis control program, the most
common causes of clinical mastitis are the environmental 756
 Mastitis Cont’d---
Source of infection and method of transmission
S. aureus is ubiquitous in the environment of dairy cattle.
The infected mammary gland of lactating cows is the major
reservoir and source of the organism.
The organism may be present on the
Skin of the teats
External orifices of heifers
Bedding materials
Feedstuffs
Housing materials
Non bovine animals on the farm and equipment
The hornfly (Hameotobia irritans) is an important vector for
transmitting S. aureus mastitis in heifers, particularly in herds with
scabs on the teat ends.
Prevention of high populations of flies in heifers is therefore 757
 Mastitis Cont’d---
needed to decrease new infections in this group.
Risk factors
As many infectious diseases, there are many risk factors that
predispose the cow to be infected by mastitis caused by S.aureus.
The risk factors can be grouped as
1. Animal risk factors
2. Environmental and management risk factors
3. Pathogen risk factors
1. Animal risk factors
Several animal risk factors influence the prevalence of infection
and the occurrence of clinical mastitis due to S. aureus.
The most important animal risk factors are
 Local defense mechanisms
Colonization with minor pathogens
 Parity of cow
 Presence of other concurrent diseases
758
 Mastitis Cont’d---
 Heredity
 Immune system
Local defense mechanisms
Abrasions of the teat orifice epithelium are an important risk factor
for S. aureus mastitis.
In experiments, teat canal infection or colonization may develop in
93% of experimentally abraded teat canal orifices compared to
53% in control quarters.
Colonization with minor pathogens
The presence of minor pathogens such as coagulase-negative
staphylococci protects against new intramammary infections
associated with the major pathogen S. aureus.
This may be the result of
An elevated SCC or
 An antimicrobial-like substance provided by the coagulase that
inhibits the growth of S. aureus.
This is not always true for all species of bacteria, because 759
 Mastitis Cont’d---
 Quarters infected with coagulase-negative staphylococci may be
more susceptible to new infections with S. agalactiae.
 Quarters that are infected with Corynebacterium bovis are
protected against S. aureus infection but not protected against
most streptococcal species.
Parity of cow
The prevalence of intramammary infection and subclinical
infection due to S. aureus increases with parity of the cow.
This is probably due to
The increased opportunity of infection with time and
 The prolonged duration of infection especially in a herd without a
mastitis control program.
Presence of other concurrent diseases
The presence of periparturient diseases has been identified as a risk
factor for mastitis.
Some of the common periparturient diseases of cattle that
760
 Mastitis Cont’d---
predispose animals to mastitis are
Dystocia
Parturient paresis
Retained placenta
ketosis
Heredity
Experimentally, the presence of certain bovine lymphocyte antigens
increased the susceptibility to S. aureus infection.
But heritability estimates of susceptibility after experimental
challenge were low and unstable.
Immune system
The infection rate of S. aureus is dependent on the ability of the
immune system to recognize and to eliminate the bacteria.
Staphylococcal antibodies are present in the blood of infected
cows.
But they appear to afford little protection against mastitis
761
associated with S. aureus.
 Mastitis Cont’d---
This may be due to the low titer of the antibodies in the milk.
However, a very crucial role in protection of mastitis caused by S.
aureus plays cellular immunity mainly polymorphonuclear
phagocytic cells.
2. Environmental and management risk factors
Several herd-level management risk factors are important for the
spread of S. aureus.
Poor teat and udder cleaning can allow spread of the organism
among quarters of the same cow, and can allow contamination of
milking units, which are commonly transferred among cows
without washing or rinsing.
Management procedures that are most effective in reducing
infection rates and cell counts associated with infections with S.
aureus are:
Postmilking teat dipping
Maintaining a good supply of dry bedding for housed cows
762
 Mastitis Cont’d---
Thorough disinfection of the teat orifice before infusing
intramammary preparations.
Milking clinical cases last
Failure to use these management techniques will increase the risk
of intramammary infection with S. aureus.
3. Pathogen risk factors
Among the pathogen risk factors, the most important one is the
virulence factors of the bacteria.
S.aureus has several virulence factors that account for its
pathogenicity and persistence in mammary tissue in spite of
adequate defense mechanisms and antimicrobial therapy.
Some of the most important virulence factors of S. aureus are
1. The presence of extracellular matrix proteins called fibronectin
and collagen which help the bacteria to
Adhere and bind to epithelial cells of the mammary gland.
 Induce the epithelial cell to internalize the organism which protects
the bacteria from both exogenous and endogenous bactericidal 763
 Mastitis Cont’d---
factors.
2. Some strains of S. aureus produce toxins
α or β toxins or
Both α and β toxins in combination
Some of the toxins may cause phagocytic dysfunction.
The beta toxin damages bovine mammary secretory epithelial cells.
β toxins increases
The damaging effects of α toxin,
 The adherence of S . aureus to mammary epithelial cells
Finally the proliferation of the organism(bacteria)
3. All strains produce coagulase (hence the term coagulase positive
S. aureus), which converts fibrinogen into fibrin; this appears to
assist the invasion of tissues.
4. Leukocidin produced by S. aureus may inactivate neutrophils.
5. Many staphylococcal strains (coagulase -positive) are able to
produce an extracellular exopolysaccharide layer surrounding the
cell wall. 764
 Mastitis Cont’d---
This capsular structure and its production of slime have been
associated with virulence against host defense mechanisms.
6.A major pathogenic factor is the ability of the organism to
colonize and produce microabscesses in the mammary gland.
So that it is protected from normal defense mechanisms, including
phagocytic activity from neutrophils.
The difficulty in removing staphylococci from an infected quarter
is due to
The bacteria's ability to survive in intracellular sites.
 There is also an ability to convert to a non susceptible L-form
when exposed to antimicrobial agents, and to return to standard
forms when the antimicrobial is withdrawn.
Economic importance
The overall prevalence of bovine mastitis due to S. aureus is much
higher than for most of pathogens that cause mastitis.
The economic consequences of bovine mastitis due to S. aureus
comprises 765
 Mastitis Cont’d---
 The need for culling of cattle out production because response to
treatment is comparatively poor
 The risk of new infections is of continuing concern.
Methods for the eradication of staphylococcal mastitis from
infected herds have yet to be devised.
Zoonotic implications
The presence of S. aureus in market milk may present a degree of
risk to the consumer.
Because of the organism's capacity to produce enterotoxins and a
toxic shock syndrome toxin(TSST), which cause serious food
poisoning.
Pathogenesis
The pathogenesis of acute and chronic S. aureus mastitis in the cow
is the same.
The variation occurring only in degree of involvement of mammary
tissue.
In both forms each focus commences with an acute stage 766
 Mastitis Cont’d---
characterized by proliferation of the bacteria in the collecting ducts
and, to a lesser extent, in the alveoli.
In acute mastitis the small ducts are quickly blocked by fibrin clots,
leading to more severe involvement of the obstructed area.
Infection during early lactation may result in the peracute form of
mastitis, with gangrene of the udder.
During the later stages of lactation or during the dry period new
infections are not usually accompanied by a systemic reaction.
But result in the chronic or acute forms.
In the gangrenous form the death of tissue is precipitated by
thrombosis of veins causing local edema and congestion of the
udder.
Strains of S. aureus are the only bacteria that commonly cause this
reaction in the udder of the cow, and the resulting toxemia is due to
bacterial toxins and tissue destruction.
Secondary invasion by E. coli and Clostridium spp. contributes to
767
 Mastitis Cont’d---
the severity of the lesion and production of gas.
In the chronic form of S.aureus mastitis there are
Fewer foci of inflammation
The reaction is milder
The inflammation is restricted to the epithelium of the ducts
This subsides within a few days and is replaced by connective tissue
proliferations around the ducts, leading to their blockage and atrophy
of the drained area.
The leukocyte infiltration into the stroma, the epithelial lining and
the lumina indicate an obvious deficiency of secretory and
synthesizing capacity due to limitation of the alveolar lumina and the
distension of the stroma area.
Clinical findings
Clinically Staphylococcus aureus mastitis can have the following
forms
1. Peracute
768
 Mastitis Cont’d---
2. Acute
3. Chronic
1. Peracute form
Peracute staphylococcal mastitis is rare but do occur and can be
fatal, even if aggressively treated.
Peracute S. aureus mastitis occurs usually in the first few days after
calving and is highly fatal.
There is a severe systemic reaction with the following clinical
features
Elevation of body temperature to 41-42°C
Rapid heart rate (100-120 beats/min)
Complete anorexia, profound depression and absence of ruminal
movements
Muscular weakness and often to the point of recumbency.
The onset of the systemic and local reactions is sudden.
The cow may be normal at one milking and recumbent and
comatose at the next. 769
 Mastitis Cont’d---
causes severe lameness on the affected side
Gangrene is a constant development in peracute form and may be
evident very early.
The followings are the main clinical features
 A bluish discoloration develops that may eventually spread to
involve the floor of the udder and the whole or part of the teat, or
may be restricted to patches on the sides and floor of the udder.
Within 24 hours the gangrenous areas become black and ooze
serum and may be accompanied by subcutaneous emphysema and
the formation of blisters.
Toxemia is profound and death usually occurs unless early,
appropriate treatment is provided.
Even with early treatment the quarter is invariably lost and the
gangrenous areas slough after 6 -7 days.
2. Acute form
Acute staphylococcal mastitis as peracute form is rare but do occur
and can be fatal, even if aggressively treated. 770
 Mastitis Cont’d---
Acute S. aureus mastitis occurs most commonly in early lactation.
The main clinical signs are
Severe swelling of the gland
The milk is purulent or contains many thick clots.
Extensive fibrosis and severe loss of function of udder always
result.
3. Chronic form
The most important dairy economic losses are caused by the chronic
form or subclinical form of mastitis.
Although 50% of cattle in a herd may be affected, only a few
animals will have abnormalities recognizable by the milker.
Many cases are characterized by a slowly developing induration and
atrophy with the occasional appearance of clots in the milk or
wateriness of the first streams.
The SCC and the CMT results of infected quarter of the milk is
increased
But the disease may go unnoticed until much of the functional 771
 Mastitis Cont’d---
capacity of the gland is lost.
The infection can persist and the disease may progress slowly over
a period of many months.
Necropsy findings
In peracute staphylococcal mastitis, the affected quarter is grossly
swollen and may contain bloodstained milk dorsally.
But only serosanguineous fluid ventrally.
There is extreme vascular engorgement and swelling, often
progressing to moist gangrene of the overlying skin.
Histologically there is coagulation necrosis of glandular
tissue and thrombosis of veins.
In milder forms of staphylococcal mastitis, the invading organisms
often elicit a granulomatous response.
Microscopically, such 'botryomycotic' cases are characterized
by granulomas with a central bacterial colony and by progressive
fibrosis of the quarter.
772
 Mastitis Cont’d---
Diagnosis
Diagnose of bovine S. aureus mastitis can be done based on
History of the herd and the farm
Clinical signs
Necropsy findings
Laboratory
In live animals subclinical forms of mastitis can be diagnosed by
using screening tests like SCC and CMT of individual cow and bulk
tank milk.
ELISA tests for detecting S. aureus antibody in milk have been
developed but are not widely used.
Bacteriological culture of milk is the best method for identifying
cows with S. aureus intramammary infection.
Problem in the laboratory identification of S. aureus is that bacteria
are shed cyclically from infected quarters.
So that a series of samples are necessary to increase overall test
773
sensitivity.
 bovine
Specimens of choice are
Milk from live animal
Chilled mammary tissue and regional lymph nodes
For histopathology fixed mammary tissue with 10 % formalin
Differential diagnosis
Because of the occurrence of the peracute form bovine S. aureus
mastitis in the first few days after parturition, the dairy producer
may conclude that the cow has parturient paresis.
But parturient paresis is characterized by weakness, recumbency,
hypothermia, rumen stasis, dilated pupils, tachycardia with weak
heart sounds.
It is easy to differentiate parturient paresis from peracute form
bovine S. aureus mastitis because
 Parturient paresis has rapid response to intravenous administration
of 10% calcium gluconate (10% calcium borogluconate).
The mammary gland is usually normal in parturient paresis.
Peracute form of bovine mastitis is characterized by tachycardia,774
 Mastitis Cont’d---
fever, weakness and evidence of severe clinical mastitis with
swelling, heat, abnormal milk with serum and blood, and sometimes
gas in the teat and often with gangrene of the teat upto the base of
the udder but not during in parturient paresis.
Other peracute bacterial types of mastitis which must be
differentiated from peracute bovine S. aureus mastitis is mastitis
caused by environmental bacteria that may cause severe systemic
reactions like E. coli and A. pyogenes.
But gangrene of the quarter is less common in mastitis caused by
E. coli and A. pyogenes.
Treatment
Treatment of S. aureus mastitis can be either intramammary
infusion or parenteral antimicrobial administration or in
combination of both methods.
However bacteriological cure rates antimicrobials do not exceed
50 % and infections commonly persist throughout the lifetime of
the cow. 775
 Mastitis Cont’d---
There are three likely reasons:
 1. Inadequate penetration of the antimicrobial agent to the site of
infection
2. Formation of L-forms of S. aureus
3. Beta-lactamase production
1. Inadequate penetration of the antimicrobial agent to the site of
infection
There is inadequate penetration of the antimicrobial agent into the
site of intramammary infection in the lactating cow.
Because the infection agent survives in phagocytes that are
inaccessible to antimicrobial agents.
2. Antimicrobial resistance
Antimicrobial resistant strains of S. aureus may occur.
They are often beta-lactamase producers.
The enzyme conferring resistance to beta -Iactam antimicrobial
agents such as penicillin G, penethamate, ampicillin and amoxicillin.
776
 Mastitis Cont’d---
First and third-generation cephalosporins and erythromycin are
effective against beta lactamase- producing staphylococci.
The first- and third-generation cephalosporins are also effective
against Gram-negative bacteria.
Therapy of bovine S. aureus mastitis can be
 1. Lactating cow therapy
 2. Dry cow therapy
1.Lactating cow therapy
The treatment of clinical cases of S. aureus mastitis using
intramammary antimicrobial infusions is less than satisfactory but is
often done.
However, clinical recovery following therapy does not necessarily
eliminate the infection.
In general, the cure rate depends on
The duration of infection
 The number of quarters infected
777
 Mastitis Cont’d---
 Antimicrobial resistance of S. aureus( beta-lactamase producer).
 The immune status of the cow
The antimicrobial agent administered
The duration of treatment.
Current recommendations to ensure the best treatment success rate
are
 To combine intramammary and parenteral antimicrobial treatment
or
To use extended intramammary treatment for 4 - 8 days.
Penicillin G is regarded as the antimicrobial agent of choice for S.
aureus strains that are penicillin-sensitive.
The following intramammary infusions, given daily at 24-hour
intervals for three treatments (unless stated otherwise) have been
used for the treatment of clinical cases of S. aureus mastitis, in
lactating cows.
Sodium cloxacillin (200-600 mg for three infusions)
778
 Mastitis Cont’d---
Tetracyclines
Penicillin-streptomycin combination
 Penicillin-tylosin combination
Novobiocin
Cephalosporins - most strains of S. aureus are sensitive to
cephapirin
 Pirlimycin-extended therapy
A slightly more effective treatment for subclinical S. aureus
intramammary infection is
 Simultaneous intramammary infusion of amoxicillin and
intramuscular injection of procaine penicillin G.
Since there is a persistence of the infection in each herd, the final
choice of the antimicrobials to be used in peracute and sub clinical
S. aureus mastitis should be based on a culture and susceptibility
test.
The application of cytokines as an adjunct to antimicrobial therapy
may help to increase the number of phagocytes in the mammary779
 Mastitis Cont’d---
gland and enhance cell function.
Experimental intramammary infusion of recombinant interleukin
into infected or uninfected mammary glands elicited an influx of
polymorphonuclear leukocytes.
This exhibits a subsequent enhanced activity and increased the cure
rate of mammary glands by 20 - 30% in quarters infected with S.
aureus.
A novel method for decreasing the transmission of S. aureus within
a herd is selectively cease lactation in infected quarters of lactating
dairy cattle.
The best method for permanently drying off a quarter is infusion of
120 ml of 5% povidone-iodine solution (0.5 % iodine) after
complete milk-out and administration of flunixin meglumine (1
mg/kg BW, intravenously).
Treatment of dairy cows with Intramammary infusions are of little
value in peracute S. aureus mastitis because of the failure of the
drugs to diffuse into the gland. 780
 Mastitis Cont’d---
But intravenous administration of large quantities of electrolyte
solutions is recommended.
Frequent massage of the udder with cold wet packs and milking out
the affected gland is recommended.
Oxytocin is used to promote milk-down but is relatively ineffective
in severely inflamed glands.
2. Dry cow therapy
It has become a common practice to leave chronic S. aureus cases
until they are dried off before attempting to eliminate the infection.
The material is infused into each gland after the last milking of the
lactation and left in situ.
The major benefits of dry cow therapy are
The elimination of existing intramammary infections and
Prevention of new intramammary infections during the dry period.
In addition, milk is not discarded and bacteriological cure rates are
superior to those obtained during lactation.
781
 Mastitis Cont’d---
Control and prevention measures
Relatively poor results are obtained in the treatment of
staphylococcal mastitis.
Any attempt to control S. aureus depend heavily on effective
methods of preventing the transmission of infection from cow to
cow.
S. aureus is a contagious pathogen and the udder is the primary site
of infection.
So that hygiene in the milking parlor is of major importance to
reduce the source of the organism.
A program of early identification, culling, and segregation is
important to control S. aureus
The control program includes:
 Hygienic washing and drying of udders before milking c regular
milking-machine maintenance.
 Teat dipping after milking(Teat dipping in 1 % iodine or 0.5%
chlorhexidine, either in 5-10 % glycerine, is completely effective
782
 Mastitis Cont’d---
against S. aureus mastitis)
 Disinfection of hands or use of rubber gloves provides additional
advantages
Dry cow treatment on all cows
Culling cows with chronic mastitis
 Milking of infected cows and quarter last (very difficult to
implement in free stall housing or pasture feeding).
1. 2. Bovine mastitis caused by Streptococcus agalactiae
Major cause of mastitis in dairy herds without an effective mastitis
control program.
Etiology
It is caused by gram positive Streptococcus agalactiae.
Epidemiology
S. agalactiae was the major cause of mastitis before the
antimicrobial era.
It is still a significant cause of chronic mastitis where control
783
procedures for contagious mastitis are not used.
 Mastitis Cont’d---
In dairy farms where there is no control measures herd prevalence
rates of infection range from 11 - 4 7%.
But in dairy farms where good hygienic measures and the efficient
treatment of clinical cases are in general use, the prevalence of
infection within a herd will be less than10 % of cows.
Source of infection
S. agalactiae is a highly contagious obligate parasite of the bovine
mammary gland.
S. agalactiae has the ability to adhere to the mammary gland tissue,
and the specific microenvironment of the udder is necessary for
growth of the organism.
The main sources of infection are
The udder of infected cows
Contaminated environment when hygiene is poor
The teats and skin of cattle
Milkers' hands
784
 Mastitis Cont’d---
Floors, utensils and clothes which are often heavily contaminated
Sores on teats are the commonest sites outside the udder for
persistence of the organism.
NB. The importance of environmental contamination as a source of
infection is given in that necessarily general disinfection technique
help to eradicate the infection.
Transmission of infection
Transmission from animal to animal occurs most commonly by
The medium of milking machine liners
 Hands
 Udder cloths
Possibly bedding
The streak canal is the portal of entry.
Risk factors
All risk factors for the occurrence of bovine mastitis caused by S.
agalactiae can be grouped
 1. Animal risk factors 785
 Mastitis Cont’d---
2. Environmental risk factors
3. Pathogen risk factors
1. Animal risk factors
There is no difference particular inbreed susceptibility.
However, when we consider the age and stage of lactation, infection
establishes more readily in older cows and in the early part of each
lactation.
The reason why older cows are more susceptible to S. agalactiae
mastitis than the younger ones is because of
Hormonal changes of cows with age
The increased hypersensitivity of mammary tissue to streptococcal
protein with age.
The physical characteristics of the teat canal may influence the
susceptibility to streptococcal infection.
Dairy cows with large teat canal lumen are more susceptible to
mastitis caused by S. agalactiae than with small teat canal lumen.
786
Because the mechanisms used by S. agalactiae to penetrate the teat
 Mastitis Cont’d---
canal are influenced more by the diameter of the teat canal lumen,
as reflected by the peak flow rate, than by teat canal length.
2. Environmental risk factors
Poor hygiene, incompetent milking personnel and machinery that is
faulty or maladjusted are important risk factors.
The most important risk factors are the failure to use post milking
teat dip and non-use of dry cow therapy.
teat dip and non-use of dry cow therapy.
The use of a common wash rag or sponge is also a risk factor.
Inadequate treatment of clinical cases of mastitis is also a frequent
risk factor in infected herds.
3. Pathogen risk factors
You know that S. agalactiae has the ability to adhere to the
mammary gland tissue, and the specific microenvironment of the
udder is necessary for growth of the organism .
The virulence of various strains of the organism is related to
differences in their ability to adhere to the mammary epithelium.787
 Mastitis Cont’d---
Economic importance
The disease is of major economic importance in intensive dairy
milk production.
In individual cows, the loss of production associated with S.
agalactiae mastitis is about 25 % during the infected lactation.
In affected herds the loss may be of the order of 10 - 15% of the
potential production.
Reduction of the productive life represents an average loss of one
lactation per cow in an affected herd.
Deaths due to mastitis by S. agalactiae infection rarely and
complete loss of productivity of a quarter is uncommon
Pathogenesis
When the bacteria have passed the primary barrier of the streak
canal and if bacteria are not flushed out by the physical act of
milking.
They proliferate and invasion of the udder tissue follows.
There is considerable variation between cows in the developments 788
 Mastitis Cont’d---
that occur at each of the three stages of invasion, infection and
inflammation.
The reasons for this variation are not clear.
After the introduction of infection into the teat, the invasion, if it
occurs, takes 1- 4 days and the appearance of inflammation 3 - 5
days
Rarely very little clinically detectable inflammation may develop
despite the persistence of a permanent bacterial flora.
The development of mastitis associated with S. agalactiae is
essentially a process of invasion and inflammation of lobules of
mammary tissue in a series of crises, particularly during the first
month after infection
Initially there is a rapid multiplication of the organism in the
lactiferous ducts and is followed by
Passage of the bacteria through the duct walls into lymphatic
vessels and to the supramammary lymph nodes.
Outpouring of neutrophils into the milk ducts. 789
 Mastitis Cont’d---
Short lived systemic reaction of the organism
Sharply falling of the milk yield as a result of inhibition and stasis
of secretion caused by damage to acinar and ductal epithelium.
The infection may results finally fibrosis of the interalveolar tissue
and involution of acini even though the tissue invasion is quickly
cleared by phagocytic cells.
Subsequently, similar crises develop and more lobules are affected
in the same way, resulting in a stepwise loss of secretory function
with increasing fibrosis of the quarter and eventually atrophy of
mammary glands.
The clinicopathological findings vary with the stage of
development of the disease.
Bacterial counts in the milk are high in the early stages
But fall when the SCC rises at the same time as swelling of the
quarter becomes apparent
The SCC rises by 10 -100 times normal during the first 2 days after
infection and returns to normal over the next 10 days. 790
 Mastitis Cont’d---
The febrile reaction is often sufficiently mild and short lived to
notice the animal.
When the inflammatory changes in the epithelial lining of the acini
and ducts begin to subside, the shedding of the lining results in the
clinical appearance of clots in the milk.
Acute swelling of udder is the combination of inflamed
interalveolar tissue and retained secretion in distended alveoli.
Removal of the retained secretion at this stage may considerably
reduce the swelling and permit better diffusion of drugs infused
into the quarter.
The inflammatory reactions also occur in the teat wall of affected
quarters.
Clinical findings
In natural cases of bovine mastitis caused by S. agalactiae there is
 Occasionally fever during initial attack which lasts for a day or
two days 791
 Mastitis Cont’d---
Inflammation of the gland persists and the subsequent crises are
usually of a relatively mild nature.
Depending on the degrees of severity bovine mastitis caused by S.
agalactiae can be classified as
1. Abnormal cow
2. Abnormal gland
3. Abnormal milk
1. Abnormal cow
When the animal is febrile, anorexic with inflamed udder and/ or
inflamed supra mammary lymph nodes.
2. Abnormal gland
When the inflammation of the gland and/ or supramammary lymph
nodes is severe but there is no marked systemic reaction.
3. Abnormal milk
When the gland is not greatly swollen, pain, heat are absent and
with out systemic reaction.
However, the presence of clots in watery foremilk may be the only
792
 Mastitis Cont’d---
apparent abnormality.
Induration is most readily palpable at the udder cistern and in the
lower part of the udder.
Necropsy findings
The gross and microscopic pathology of mastitis associated with S.
agalactiae have no importance in the diagnosis of the disease.
Diagnosis
Diagnosis of bovine mastitis can be done based on
History of the farm and dairy farms
Clinical signs (only during abnormal cow and abnormal glands
cases)
Physical examination of milk
Laboratory
Laboratory method can be made based on
1. SCC on milk samples from individual cows or quarters.
S. agalactiae produces high SCC in individual cows, which has a
significant influence on the bulk tank milk SCC. 793
 Mastitis Cont’d---
2. Isolation of the bacteria and determination of total bacterial
count from bulk tank and individual cow milk samples
Due to the presence of S. agalactiae mastitis in the herd, the total
bacterial count in bulk tank milk can be markedly increased.
Samples of bulk tank milk from infected herds commonly contain
bacterial counts in the range of 20 000-100 000 CFU/ml
Because a cow in the early stages of infection can shed upto 100
000 000 bacteria/ml.
3. Identification of the bacteria from bulk tank and individual
cow milk samples
You know in microbiology after isolation of the pure culture of S.
agalactiae, the bacteria is identified after Lancefield grouping
using CAMP test.
That is why the CAMP test, which has served as the universally
used means of identifying S. agalactiae for many years.
Nowadays, CAMP test has been replaced by a commercial latex
794
agglutination.
 Mastitis Cont’d---
Differential diagnosis
Differentiation from other types of acute and chronic mastitis is not
possible clinically.
So that the clinical diagnosis of bovine S. agalactiae mastitis
depends entirely on the isolation of S. agalactiae from the milk.
Treatment
S. agalactiae is very sensitive to intramammary therapy using a
wide variety of commercially available intramammary infusion
preparations.
Systemic therapy must be given if there is a sign of systemic
infection(abnormal cow).
Clinical and subclinical bovine S. agalactiae mastitis identified at
any stage of lactation should be treated immediately because
 It helps us to prevent the transmission of the agent to uninfected
quarters.
The disease has an excellent response to treatment.
795
 Mastitis Cont’d---
It is possible to treat bovine S. agalactiae mastitis at all stages of
lactation with antimicrobials like
Penicillin ( the most effective one is procaine penicillin)
Erythromycin
Tylosin
Cloxacillin
Cephalosporins
All of them give 90 -100% cure rates.
To provide a broader spectrum of antimicrobial efficiency penicillin
is often combined with other drugs that are more effective against
Gram-negative organisms.
 A mixture of penicillin (100 000 units) and novobiocin (150 mg)
provides a cure rate ranging from 89 - 98 %.
In clinical bovine S. agalactiae mastitis three infusions at intervals
of 24 hours are recommended.
As a general rule clinical cases of bovine S. agalactiae mastitis
should be treated with three infusions 796
 Mastitis Cont’d---
While subclinical cases of bovine S. agalactiae mastitis, particularly
those detected by routine examination in a control program should
be treated with one intramammary infusion.
Other antimicrobial agents used in the treatment of S. agalactiae
infections include
Tetracyclines
Cephalothin
Lincomycin
Neomycin
They are as effective as penicillin.
In addition, they have an advantage over penicillin in that they have
a wider antibacterial spectrum.
So that they have an obvious advantage when the type of infection is
unknown and mixed.
Lincomycin (200 mg) combined with neomycin (286 mg) and
administered twice at 12-hour intervals also has good efficacy.
797
 Mastitis Cont’d---
NB. If therapy is done during dry cows period one infusion of
antimicrobial infusion is sufficient.
Prevention and control measures
Bovine S. agalactiae mastitis can be controlled in intensive dairy
farms by
 Suitable hygienic barriers against infection
 Elimination of the agent from individual quarters by treatment
since the disease is eradicable fairly simply and economically.
The control program consists of
Identifying infected quarters
Treating infected quarters on two occasions ( immediate and dry
cow therapy) if necessary
Culling incurable cows
Bovine S. agalactiae mastitis can be prevented by performing
Premilking teat and udder sanitation
Postmilking teat dipping
Dry cow therapy 798
 Mastitis Cont’d---
By having strong biosecurity rules in your farm (introduction of non
- infected animals, even heifers that have not yet calved, or the
employment of milkers who do not carry infection).
1.3. Bovine mastitis caused by Mycoplasma bovis and other
Mycoplasma species
It is also a contagious mastitis which can spread from infected
quarters to other quarters and from infected cows to healthy herds.
Etiology
It is caused mainly by Mycoplasma bovis.
Occasionally by other Mycoplasma species like
Mycoplasma F-38
Mycoplasma arginini
Mycoplasma bovirhinis
Mycoplasma canadensis
Mycoplasma bovigenitalium
Mycoplasma alkalescens
 Mycoplasma capricolium 799
 Mastitis Cont’d---
Mycoplasma californium
Mycoplasma dispar
However, M. bovis remains the most common cause of
mycoplasmal mastitis in dairy cattle.
Epidemiology
A highly contagious mastitis causing outbreaks of clinical mastitis.
The disease is mostly common in large herds with recent
introductions.
Transmitted within herds by bulk mastitis treatments and poor
milking hygiene.
The disease affects cows of all ages and any stage of lactation.
But those in early lactation are most severely affected.
Risk factors
Cows of all ages and at any stage of lactation are affected.
Cows that have recently calved showing the most severe signs and
dry cows the least.
There are several recorded outbreaks in dairy herds in dry cows 8.00
 Mastitis Cont’d---
One of them immediately after mammary infusions of dry period
treatment and all quarters of all cows are affected.
Economic importance
The disease is a disastrous one.
Because of the high incidence in affected herds and
The almost complete cessation of production for the lactation.
Many cows fail ever to return to milking; as many as 75 % of
affected cows may have to be culled.
Pathogenesis
The bacteria cause purulent interstitial mastitis.
Although infection probably occurs via the streak canal, the rapid
spread of the disease to other quarters of the udder and occasionally
to joints suggests indicates that the bacteria can spread
hematogenous.
The presence of the infection in heifers milked for the first time
also suggests that systemic invasion may be followed by
localization in the udder. 801
 Mastitis Cont’d---
Clinical findings
It causes clinical, subclinical and chronic form of intramammary
infections in cattle.
In lactating cows, the main clinical signs are
A sudden onset of swelling of the udder except dry cows which
show little swelling of the udder
In most cases all four quarters are affected without obvious signs of
systemic illness
A sharp drop in milk production specially high -producing cow may
fall in yield to almost nil between one milking and the next.
There is no overt evidence of systemic illness, and febrile reactions
in most field cases in lactating cows.
But those that have recently calved show most obvious swelling of
the udder and may be off their feed( anorexia) and have a mild
fever.
Secretion in affected quarters at the early stage is in that it appears
fairly normal at collection. 802
 Mastitis Cont’d---
But on standing a deposit, may form of fine, sandy material,
flakes or floccules, settles out leaving a turbid whey-like
‘supernatant.
Eventually there is atrophy of udders which do not return to
production.
Calves suckling milk from infected cows may develop otitis
media/interna.
In some cases the supramammary lymph nodes are greatly enlarged.
NB. The classic clinical presentation in bovine mastitis caused by
M. bovis and other species of Mycoplasma is severe clinical mastitis
in multiple quarters of multiple cows with minimal systemic signs
of disease.
From all mycoplasmal bovine mastitis, bovine mastitis due to M.
bovigenitalium may be very mild.
It disappears from the herd spontaneously and without causing loss
of milk production.
803
 Mastitis Cont’d---
Necropsy findings
The main necropsy findings that are observed in bovine
mycoplasmal mastitis are
 The presence of grossly, diffuse fibrosis and granulomatous lesions
containing pus in the mammary tissue.
 The thickening and roughening of lining of the milk ducts and the
teat sinus.
 On histological examination the granulomatous nature of the
lesions is evident.
Metastatic pulmonary lesions have been found in a few long-
standing cases.
Diagnosis
Diagnose of bovine mycoplasmal mastitis can be made based on
History of the farm
Clinical signs
SCC using CMT
804
 Mastitis Cont’d---
Laboratory
Diagnosis at the herd level can be made by culturing bulk tank milk
or milk from cows with clinical mastitis.
It is also possible to determine somatic cell counts(SCC) in the
milk by using CMT and SCC is very high, usually over 20 000
000 cells/ml.
The diagnose can be confirmed by isolation and identification of
the organism in the laboratory.
The appropriate specimens are
Milk in acute stage
Chilled mammary tissue, and regional lymph node
Formalin fixed mammary tissue for histopathology
Samples for culture should be freshly collected and transported at
4°C.
The causative organism can be cultured without great difficulty by
a laboratory skilled in working with Mycoplasma in special media
called beef infusion with special supplement. 805
 Mastitis Cont’d---
In the acute stages the organisms may be able to be visualized by
the examination of a milk film stained with Giemsa or Wright
Leishman stain.
Species identification of Mycoplasma isolates is most commonly
done using
Immunofluorescence
Homologous fluorescein-conjugated antibody or
An indirect immunoperoxidase test (immunohistochemistry).
Differential diagnosis
A presumptive diagnosis can be made based on the clinical
findings.
But laboratory confirmation by culture of the organism is desirable.
The facts that the organism does not grow on standard media.
Other pathogenic bacteria are commonly present often lead to
errors in the laboratory diagnosis unless attention is drawn to the
characteristic field findings.
806
 Mastitis Cont’d---
Treatment
Cows diagnosed with mycoplasmal mastitis should be considered to
be infected for life.
None of the commonly used antimicrobial agents appear to be
effective and oil-water emulsions used as intramammary infusions
appear to increase the severity of the disease.
Parenteral treatment with oxytetracycline (5 g intravenously, daily
for 3 days) has been shown to cause only temporary improvement.
A mixture of tylosin 500 mg and tetracycline 450 mg used as an
infusion cured some quarters.
Unless treatment is administered very early in the course of the
disease, the tissue damage has already been done.
Control and preventation measures
Affected animals should be culled immediately or placed in strict
isolation until sale for meat.
Eradication of the disease can be achieved by culling infected cows
identified by culture of milk and nasal swabs, especially at drying
807
 Mastitis Cont’d---
off and calving.
When eradication is completed the bulk tank milk SCC is the best
single monitoring device to guard against reinfection.
An alternative program recommended for large herds is the
creation of an infected sub herd that is milked last.
Cows with infected quarters are segregated into the sub herd.
Intramammary infusions must be carried out with great attention to
hygiene and preferably with individual tubes rather than multi dose
syringes.
Most commercial teat dips are effective in control.
Use of disposable latex gloves with disinfection of the gloved
hands between cows may minimize transmission at milking.
Because M. bovis can cause respiratory disease, otitis media/intema
and arthritis in calves, all colostrum and waste milk fed to calves
should be pasteurized.

808
 Mastitis Cont’d---
1. 4. Bovine mastitis caused by Corynebacterium bovis
It is also a contagious bovine mastitis which is commonly
associated with subclinical infection.
Etiology
It is caused by gram positive pleomorphic bacteria called
Corynebacterium bovis.
It is a common and very contagious pathogen.
There is considerable debate about the significance of C. bovis
infections for mammary gland health and cow productivity.
For this reason, C. bovis is classified as a minor pathogen of
bovine mastitis.
Epidemiology
The main reservoir of infection appears to be infected glands and
teat ducts.
C. bovis spreads rapidly from cow to cow in the absence of
adequate teat dipping.
809
 Mastitis Cont’d---
C. bovis is extremely contagious and the duration of intramammary
infection is long (many months).
The prevalence of C. bovis bovine mastitis is typically low in herds
where an effective germicidal teat dip, good milking hygiene and
dry cow therapy are practiced.
Many studies demonstrated that the bacteria has a predilection for
the streak canal.
Clinical findings
An intramammary infection with C. bovis is infrequently associated
with clinical disease .
But it usually causes a moderate increase in the SCC and a small
increase in the CMT score.
The main clinical signs are
Milk production losses are usually not detectable
 Mastitic milk secretion is typically a thicker than normal milk
(abnormal milk)
Occasional cases may have a large firm gland (abnormal gland). 810
 Mastitis Cont’d---
Treatment
C. bovis is very susceptible to penicillin, ampicillin, amoxicillin,
cephapirin, and erythromycin.
So that it is possible to use commercially available intramammary
infusions that contains the above antibiotics.
Since the bacteria do not cause systemic infection, no need for
parenteral treatment.
The duration of infection can be prolonged (months) if the animals
not treated with antimicrobial agents in time.
Preventation and control measures
Long-term intensive programs of teat dipping and dry cow therapy
will markedly reduce the prevalence of bovine mastitis caused by C.
bovis.

811
 Mastitis Cont’d---
2. Mastitis of cattle associated with common environmental
pathogens
 2.1. Bovine coliform mastitis
 2.2. Bovine mastitis caused by environmental streptococci
 2.3. Bovine mastitis caused by Arcanobacterium pyogenes
Environmental bovine mastitis is associated with bacteria that are
transferred from the environment to the cow rather than from other
infected quarters and/ or cows.
The main environmental pathogens that cause environmental bovine
mastitis includes
 1.Coliform mastitis associated with Escherichia coli, Klebsiella
species and Enterobacter aerogenes
 2. Environmental streptococci
 3. Arcanobacterium pyogenes
2.1. Coliform mastitis associated with Escherichia coli, Klebsiella
species and Enterobacter aerogenes
It is environmental bovine mastitis caused by coliform
812
Enterobactericeae(lactose fermenter).
 Mastitis Cont’d---
Etiology
Many different serotypes of E.coli, numerous capsular types of
Klebsiella spp. (most commonly K. pneumoniae) and Enterobacter
aerogenes are responsible for coliform mastitis in cattle.
Other Gram-negative bacteria in the family of Enterobactericeae
which are not categorized as coliforms but can cause mastitis
include
Serratia spp.
Pseudomonas spp.
Proteus spp.
Epidemiology
Occurrence of coliform mastitis
Coliform mastitis occurs worldwide.
The occurrence of coliform mastitis has increased considerably in
recent years.
Because of intensification of dairy farms.
It is mostly common in dairy cattle that are housed in total 813
 Mastitis Cont’d---
confinement during the winter or summer months.
Clinical mastitis associated with environmental pathogens
(including the environmental streptococci) is now the most
important mastitis problem in intensive dairy farms.
However, the disease is uncommon in dairy cattle that are
continuously in pasture.
In contrast to contagious mastitis, environmental mastitis associated
with coliform bacteria is primarily associated with clinical mastitis
rather than subclinical mastitis.
Coliform intramammary infections are usually short duration.
 82 – 90 % of coliform mastitis have acute form
 8-10% of coliform infections result in peracute mastitis
Source of infection and mode of transmission
The primary reservoir of coliform infection is the dairy cow's
environment (environmental pathogen).
This is in contrast to the infected mammary gland, which is the
reservoir of major contagious pathogens (S. aureus and S. 814
 Mastitis Cont’d---
agalactiae) and the main reservoir of infection in cattle with M. bovis.
Exposure of uninfected quarters to environmental pathogens can
occur at any time during the life of the cow, including during
 During milking
 Between milkings
During dry period
Before calving in heifers.
Coliform mastitis is one of the most common causes of fatal mastitis
in cattle.
The case fatality rate from peracute coliform mastitis is commonly
high and may reach 80% in spite of intensive therapy.
Outbreaks of the disease can occur with morbidity upto 25% of
recently calved cows.
Risk factors
The main risk factors for the occurrence of environmental mastitis
are
1. Pathogen risk factors 815
 Mastitis Cont’d---
 2. Environmental risk factors
 3. Animal risk factors
1. Pathogen risk factors
The pathogen risk factors are
LPS(endotoxin)
Somatic and capsular factors to form resistance to bovine
bactericidal activity of serum
The fibronectin binding property of E. coli from bovine mastitis
may be an important virulence factor that allows the organism to
adhere to the ductular epithelium.
2. Environmental risk factors
All the environmental components that come in contact with the
udder of the cow are considered potential sources of the organisms.
The coliform bacteria are opportunists and contamination of the
skin of the udder and teats occurs primarily between milkings.
Contamination of the udder and the teats with coliform bacteria
816
 Mastitis Cont’d---
may occur
When the cow is in contact with contaminated bedding rather than at
the time of milking.
 Feces, which are a common source of E. coli, can contaminate the
perineum and the udder directly or indirectly through bedding,
calving stalls, dry lot grounds, udder wash water, udder wash
sponges and cloth rags, teat cups and milkers' hands.
 Direct transmission probably occurs through the milking machine.
Environmental risk factors that affect the occurrence of coliform
mastitis are bedding materials.
The different materials that can be used as bedding material in dairy
farms are
 Sawdust
 Shavings
 Straw
Many studies showed that cows bedded on sawdust have the largest
817
teat end population of total coliforms and Klebsiella
 Mastitis Cont’d---
Those bedded on shavings have an intermediate number
While those on straw have the least number
Wet bedding, particularly sawdust and shavings, promotes the
growth of coliform bacteria, especially Klebsiella spp.
3. Animal risk factors
Animal risk factors that influence the susceptibility of cows to
coliform mastitis include
The SCC of the milk
Neutrophil recruitment and function
Selenium and vitamin E status
Stage of lactation and defense mechanism
Contamination of teat duct
Downer cows
Other defense factors(lactoferrin and citrate)
Somatic cell counts of 500 000 cells/ml provided complete
protection.
818
 Mastitis Cont’d---
Thus cows in herds with a low incidence of streptococcal and
staphylococcal mastitis have a low milk SCC and are more
susceptible to coliform mastitis.
Increased susceptibility to coliform mastitis in the periparturient
cow is primarily due to impaired neutrophil recruitment to the
infected gland.
In fatal cases of peracute mastitis in cows within 1 week after
parturition there may be large numbers of bacteria in mammary
tissues and an absence of neutrophilic infiltration.
The lack of neutrophil mobilization could be due to
Failure to recognize bacteria
Lack of production of inflammatory mediators
A defect in the ability of the cells to move into the milk
compartment.
An adequate dietary level of selenium enhances the resistance of
the bovine mammary gland to infectious agents.
The enhanced resistance is thought to be associated with a more819
 Mastitis Cont’d---
rapid diapedesis of neutrophils into the gland of cows fed diets
supplemented with selenium, which limits the numbers of bacteria
in the gland during infection
Vitamin E is especially important to mammary gland health during
the peripartum period.
Plasma concentrations of alpha-tocopherol begin to decline at 7 - 10
days before parturition and at 3-5 days after calving.
When plasma concentrations are maintained during the peripartum
period by injections of alpha-tocopherol, the killing ability of blood
neutrophils is improved.
The supplementation of the diets of dry cows receiving 0.1 PPM of
selenium in their diets with vitamin E at dose of 4000IU/ day for 14
days in dry period reduces mastitis by 90%.
Coliform mastitis occurs almost entirely in the lactating cow and
rarely in the dry cow.
The difference in the susceptibility may be due to the much higher
SCCs and lactoferrin concentrations in the secretion of dry quarters
820
 Mastitis Cont’d---
than in the milk of lactating quarters.
The sporadic occurrence of the disease may be associated with the
use of contaminated teat siphons and mastitis tubes and infection
following traumatic injury to teats or following teat surgery.
Cows affected with the downer cow syndrome following parturient
paresis, or recently calved cows that are clinically recumbent for
any reason, are susceptible to coliform mastitis .
This is because of the gross contamination of the udder and teats
with feces and bedding.
The failure of lactoferrin within mammary secretions to prevent
new infections.
Mastitis near and after parturition may be due to a decrease in
lactoferrin before parturition.
Lactoferrin normally binds the iron needed by iron-dependent
organisms.
These multiply excessively in the absence of lactoferrin
Citrate concentrations increase in mammary secretions at parturition
and may interfere with iron-binding by lactoferrin.
821
 Mastitis Cont’d---
Pathogenesis
After invasion and infection of the mammary gland, E. coli
proliferates in large numbers and releases endotoxin on bacterial
death.
Endotoxin of E. coli causes
 A change in vascular permeability, resulting in edema and acute
swelling of the gland .
 A marked increase in the number of neutrophils in the milk.
The neutrophil concentrations may increase 40 - 250 times and
strongly inhibit the survival of E. coli and others that cause
coliform mastitis.
This marked diapedesis of neutrophils is associated with the
remarkable systemic leukopenia and neutropenia that occurs in
peracute coliform mastitis.
The severity of coliform mastitis is influenced by
The degree of the pre-existing neutrophils in the milk
 The rate of invasion of bacteria and total number of neutrophils 822
 Mastitis Cont’d---
that invade the infected gland.
Susceptibility of the bacteria to serum bactericidins that are secreted
into the gland
The amount of endotoxin produced.
Many biological active substances are produced during coliform
mastitis.
The most common biological active substances that are produced
during coliform mastitis are
Histamine
Serotonin
Leukotriene
Arachidonic metabolites like prostaglandins
This indicates that they may play a role in the pathogenesis of
endotoxin-induced mastitis.
So that the use of NSAIDs has a great value in therapeutics of
coliform mastitis.
823
 --
Mastitis Cont’d-
In peracute form of coliform mastitis, there is
Severe toxemia
 Fever
Shivering
Weakness leading to recumbency in a few hours
Mild diarrhea
These are probably due to
 Absorption of large quantities of endotoxin from the site of
inflammation(udder)
Bacteremia which is present in 32 - 48% of naturally occurring
cases of coliform mastitis.
Endotoxemia in cattle due to coliform mastitis cause
Profound systemic leukopenia (neutropenia and lymphopenia)
A mild hypocalcemia
Elevation in plasma cortisol concentration.
Profound systemic leukopenia (neutropenia and lymphopenia) is
because of a marked diapedesis of neutrophils to the site of 824
 Mastitis Cont’d---
inflammation.
Hypocalcemia that occurs in naturally occurring cases and is thought
to be due to
Decreased abomasal emptying rate(hypotony and atony of GIT))
associated with the endotoxemia.
Increased plasma cortisol concentration which inhibits Ca ion
absorvation.
In the acute form, the systemic changes are usually less severe than
in the peracute form.
However, in both forms,
There is marked agalactia
 Secretions in the affected quarter become serous and contain small
flakes.
Clinical findings
Coliform mastitis can be classified
 1. Clinical coliform mastitis
 2. Subclinical coliform mastitis 825
 Mastitis Cont’d---
1. Clinical coliform mastitis
Clinical coliform mastitis can be
Peracute
Acute
Subacute
Chronic
Peracute coliform mastitis in the cow is a severe disease
characterized by a sudden onset of agalactia and toxemia.
The peracute form of the disease is most common in recently
calved cows.
The cow may be normal at one milking and acutely ill at the next.
The main common clinical signs in peracute coliform mastitis are
 Anorexia
Severe depression
 Shivering and trembling
 Cold extremities (particularly the ears)
826
 Mastitis Cont’d---
 Fever of 40 - 42°C
Within 6 - 8 hours after the onset of signs the cow may be
recumbent and unable to stand.
At that stage, the temperature may be normal or subnormal all of
which may superficially resemble parturient paresis.
The heart rate is usually increased upto 100-120 beats/min.
The rumen is static( atony of rumen)
There may be a mild watery diarrhea and dehydration
Polypnea is common and in severe cases an expiratory grunt may be
audible because of pulmonary congestion and edema.
 The affected quarter(s) is usually swollen and warm but not
remarkably
So that coliform mastitis may be missed on initial clinical
examination.
 The cow may be severely toxemic, febrile and have cold
extremities before visible changes are occurring in the mammary
gland or the milk. 827
 Mastitis Cont’d---
 The mammary secretion is characteristic, and changes from th
consistency of watery milk initially to a thin, yellow serous fluid
containing small meal-like flakes that are barely visible to the naked
eye.
Additional quarters may become affected within a day or two of the
initial infection.
The course of peracute coliform mastitis is rapid.
Some cows will die in 6 - 8 hours after the onset of signs
Others will live for 24 - 48 hours.
Those that survive the peracute crisis will either
Return to normal in a few days or
Remain weak and recumbent for several days and eventually
develop the complications associated with prolonged recumbency.
Intensive intravenous fluid therapy may prolong the life of the cow
for up to several days.
But significant improvement may not occur and eventually
euthanasia may appear to be the desirable course of action. 828
 Mastitis Cont’d---
Acute coliform mastitis is characterized by
Varying degrees of swelling of the affected gland
 The secretions of the gland are watery to serous in consistency and
contain flakes.
Acute form of coliform mastitis differs from peracute form in that
recovery with appropriate treatment usually occurs in a few days.
Subacute coliform mastitis has similar clinical signs like acute
mastitis but less sever.
Chronic coliform mastitis is characterized by repeated episodes of
subacute mastitis.
So it cannot be readily clinically distinguished from other common
causes of mastitis.
2. Subclinical coliform mastitis
Subclinical coliform mastitis is characterized by the presence of
coliform organisms in the milk samples of cows without clinical
evidence of mastitis.
829
 Mastitis Cont’d---
The prevalence subclinical coliform mastitis ranging from 0.9 -
1.2% from the total cause of subclinical mastitis.
Necropsy findings
There is edema and hyperemia of the mammary tissue.
 In severe cases hemorrhages are present and are accompanied by
thrombus formation in the blood and lymphatic vessels
There is necrosis of the parenchyma.
Tissue damage is most marked in the epithelium of the teat and
lactiferous sinuses
This damage diminishes rapidly towards the ducts.
In peracute cases, the organisms are largely confined to the ductular
and secretory lumen.
 In some cases there may be intense neutrophil infiltration, sub
epithelial edema and epithelial hyperplasia of the sinuses and large
ducts
 There is now some evidence that bacteremia may occur in coliform
mastitis. 830
 Mastitis Cont’d---
Diagnosis of coliform mastitis
Diagnose of coliform mastitis is done based on
History of the animals and the actual situation of the farm
Clinical signs
SCCs and CMT
Laboratory including hematology, endotoxin detection,and bacteria
isolation and identification.
In the actual situation of the farm, you must consider the hygienic
condition.
Coliform mastitis has mostly acute and peracute forms with less
subclinical form.
In coliform mastitis, the SCCs of affected quarters ranges 14 000
000 - 25 000 000 cells/ml.
The CMT on secretions from affected quarters is usually +3.
Hematologically, in peracute coliform mastitis there is
An increased PCV value (haemoconcentration)
A marked leukopenia, neutropenia and a degenerative left shift due
831
 Mastitis Cont’d---
to the margination of large numbers of neutrophils in response to
endotoxin.
A moderate lymphopenia, monocytopenia and thrombocytopenia.
Endotoxin detection also has very important value to diagnose
coliform mastitis.
Endotoxin can be detected in the milk and/or plasma of affected
animals.
A commercially available cow side test called Limast-test for
endotoxin is available.
The test takes 15 minutes to run on milk samples and is able to
detect the presence of endotoxins of Gram negative bacteria.
But it does not differentiate between E. coli, K. pneumoniae and
other Gram negative bacteria that cause coliform mastitis.
Laboratory diagnosis also help to isolate, identify the bacteria and
to do antibiotic sensitivity test.
Samples for confirmation of diagnosis can be from live and dead
animals 832
 Mastitis Cont’d---
From live animals milk sample is the appropriate sample to isolate
the bacteria.
From dead animals chilled mammary tissue and regional lymph
node can be collected for bacteria isolation and identification
For histopathology formalin-fixed mammary tissue can be used
Milk samples should be submitted for culture to identify the
causative agent.
In the peracute case, the milk samples will yield a positive culture.
In less acute cases, the milk sample may be negative because the
neutrophils have cleared the bacteria.
Differential diagnosis
Peracute coliform mastitis must be differentiated from
Parturient paresis (milk fever due to hypocalcemia)
Acute carbohydrate engorgement called rumen lactic acidosis
The weakness and recumbency because of parturient paresis
resembles peracute coliform mastitis.
But parturient paresis does not has a marked increase in heart 833
 Mastitis Cont’d---
rate, dehydration and mild diarrhea.
In peracute coliform mastitis shows clinical signs of fever, watery
consistency of milk, shivering, firmness of udder, tachycardia and
polypnea but not in parturient paresis.
Peracute coliform mastitis shows a marked leukopenia and
neutropenia
Whereas in parturient paresis there is usually a neutrophilia and
stress leukon (neutrophilia, no left shift, lymphopenia, monocytosis
and eosinopenia).
Other diseases of cattle that must be differentiated from peracute
mastitis is acute carbohydrate engorgement called rumen lactic
acidosis .
Because it causes rapid onset of weakness, recumbency, diarrhea,
dehydration and ruminal stasis which resembles the clinical
findings of shock in peracute coliform mastitis.
However, it is simple to differentiate them because during acute
834
CH2O engorgement
 Mastitis Cont’d---
The rumen contains an excess of watery fluid and
The PH of rumen content is below 5
Acute coliform mastitis cannot be accurately differentiated from all
other common causes of acute mastitis with abnormal gland and
abnormal milk.
The most common pathogens that can cause acute mastitis and
that must be differentiated from acute coliform mastitis are
Environmental streptococci (S. uberis and S. dysgalactiae)
Contagious pathogens (S. aureus and S. agalactiae).
They can be differentiated by isolation and identification the
bacteria.
Treatment
In generally the following combined methods of treatments are
more effective to treat peracute and acute coliform mastitis that
cause systemic sign of the disease(abnormal cow).
 Broad spectrum antimicrobial agents administered parenterally
and by intramammary infusion. 835
 Mastitis Cont’d---
Intravenous fluid and electrolyte therapy to prevent dehydration
 Frequent stripping out of the affected glands with the aid of
oxytocin and mechanical hand massages
Using of anti inflammatory drugs to control inflammation which is
caused by endotoxin of the bacteria(E. coli).
The major considerations for antimicrobial use to treat coliform
mastitis include:
Early administration in order to decrease the exposure of the cow to
endotoxin
 Ensuring appropriate withholding periods for milk and meat
The benefit-cost ratio
Broad-spectrum antimicrobial agents should be administered
parenterally to cattle with systemic signs of disease (abnormal
cow) in addition to intramammary infusion.
It is preferably to administer by the intravenous route initially
followed by intramuscular administration to maintain the
836
appropriate plasma concentrations.
 Mastitis Cont’d---
The main reason to administer parenteral antibiotics is to combat
bacteremia which is present in 32 - 48% of severely affected cattle
with coliform mastitis.
Experimentally most E. coli isolated from the mammary glands of
cattle are theoretically susceptible to
Third-generation cephalosporins (such as ceftiofur)
 Fourth-generation cephalosporins (such as Cefquinome)
Neutrofurans like fluoroquinolones
 Amikacin(antibiotic)
Sulfa drugs like trimethoprim-sulfonamide
Oxytetracycline.
So that the above antimicrobial drugs can be administered
parenterally and/or intramammary in the form of infusion.
Parenteral administration
Ceftiofur can be administered IM in dose of 2.2 mg/kg BW every
24 h for three days
Cefquinome can be administered IM in dose of 1 mg/kg BW two 837
 Mastitis Cont’d---
times at 24 h apart with or without intramammary infusion.
Cefquinome can be administered intramammary at dose of 75 mg,
three times at 12 h intervals is effective in coliform mastitis.
Antimicrobial drugs including neutrofurans and sulfa drugs are also
effective in coliform mastitis.
Parenterally administration of neutrofurans like fluoroquinolones
and Enrofloxacin are effective.
Sulfa drugs like trimethoprim-sulfadiazine (trimethoprim 4 g,
sulfadiazine 20 g) given intramuscularly every 24 h for 3 - 5 days
is effective to treat naturally acquired cases of coliform mastitis.
Sulfa drugs are more effective when we are using in combination
with NSAlDs and complete milking of affected quarters several
times daily.
Oxytetracycline (16.5 mg/kg BW intravenously every 24 hours for
3-5 days), combined with intramammary cephapirin (200 mg) and
supportive care (intravenous or oral fluids, flunixin meglumine,
stripping of the mammary gland) is effective in treatment of 838
 Mastitis Cont’d---
coliform mastitis.
Intramammary antimicrobial agents
Intramammary preparations of antimicrobial agents can be infused
into the affected quarters after they have been stripped out
completely at the start and end of the day.
The most common commercially available intramammary
antimicrobial agents are
Ceftiofur : - an excellent choice for intramammary infusion in
suspected cases of coliform mastitis.
It is administered at the dose of 200 mg/ quarter very 12 hours for
two milkings.
If the intramammary infusion of ceftiofur is combined with IM
administration of 100 units of oxytocin every 12 hours immediately
before milking for two or three milkings, it has short-term clinical
or bacteriological cure rates.
Amoxicillin: - It is also every an excellent choice for
intramammary infusion in suspected cases of coliform mastitis at839
 Mastitis Cont’d---
dose of 62.5 gm/ quarter every 12 hours for three milkings.
Stripping of affected quarter
Many studies showed that frequent stripping of the udder would be
an effective treatment.
Using of oxytocin at the dose of 10 - 20 units per adult cow given
intramuscularly, followed by vigorous hand massage and hourly
stripping of the affected quarter, may assist in removing
inflammatory debris.
Effective removal of coliform bacteria and endotoxin will
minimize their local effects in the mammary gland and decrease
the systemic signs of endotoxemia.
It is known that the main problems with stripping are
 Labor involved
 Potential for creating additional pain and discomfort for the cow
(and the producer when the cow kicks!)
 Potential contamination of the environment if the secretion is
stripped on to the ground. 840
 Mastitis Cont’d---
However, many studies showed that the role of frequent stripping, if
any, in the treatment of clinical mastitis remains to be determined.
Fluid and electrolyte therapy
Fluid and electrolyte therapy are essential for the treatment of acute
and peracute coliform mastitis.
It helps to counteract the effects of the endotoxemia.
Fluids and electrolytes that are commonly used to overcome
endotoxemia during coliform mastitis are
Isotonic polyionic electrolyte solutions (such as Ringer's solution)
Alternative it is possible to use small volumes of hypertonic saline,
which can and administered rapidly.
The isotonic polyionic solution is given at dose of 80 ml/kg BW
for the first 24 hours by continuous intravenous infusion(give 50%
of the solution in the first 4 hours then the remains in the next 20
hours).
Hypertonic saline (7.2% NaCl) is given intravenously at 4-5 ml/kg
841
BW intravenously over 4 - 5 minutes followed by immediate access
 Mastitis Cont’d---
to drinking water.
Anti-inflammatory agents
NSAIDs are frequently administered as supportive therapy in
coliform mastitis, particularly in the peracute form.
The most common NASIDs that are commonly used singly or in
combination to treat peracute coliform mastitis are
Ketoprofen
Phenylbutazone and dipyrone
Flunixin meglumine or dexamethasone
Long acting NSAIDs called Carprofen
Carprofen is long acting NSAID which can reduce fever,
tachycardia and udder swelling associated in E. coli endotoxin-
induced mastitis.
NB.Using of combination therapy (antimicrobial, anti-
inflammatory, fluid and electrolyte therapy and stripping of the
udder) in peracute and acute coliform mastitis have good
therapeutic result. 842
 Mastitis Cont’d---
Control and preventation measures
The control and preventation measures of coliform mastitis is
characteristically difficult, unreliable and frustrating.
The general principles of mastitis control that have been effective for
the control of contagious mastitis( bovine mastitis caused by S.
aureus and S. agalactiae) have been unsuccessful for the control of
coliform mastitis.
Because infection of the mammary gland occurs by direct contact
with the environment usually between milkings.
So the following practices help us to prevent and control coliform
mastitis.
1. Management of outbreaks
It includes:
 Culture milk samples and obtain a definitive etiological diagnosis
(in other words, put a name to the causative pathogen).
 Examine the bedding for evidence of heavy contamination with
843
 Mastitis Cont’d---
coliform bacteria(if sawdust or wood shavings are being used,
replace with sand or straw if possible, or change more frequently)
 Conduct a general clean-up of the stall and lounging areas
 Improve premilking hygiene
 Examine milking machine function
 Allow cows access to fresh feed immediately after milking to
ensure that they remain standing for at least 30 minutes to allow
time for the streak canal to close.
2. Housing and environment ( Bedding, regular daily cleaning
of barns)
The overall level of sanitation and hygiene must be improved and
maintained in the herd.
Since most coliform bovine mastitis occur in periparturient period
( very early in the dry period or just before calving), much efforts
to prevent the infection should be centered on these periods.
So that management of the dry cow environment may provide the
best opportunity for prevention of infection. 844
 Mastitis Cont’d---
Cows that are housed during part or all of the day or night should be
bedded on clean and dry bedding and not overcrowded, to prevent
heavy fecal contamination.
That means bedding should be kept as dry as possible.
The use of sawdust or shavings as bedding should be avoided if
possible.
Sand is now considered to be the ' gold standard' and the most
suitable alternative because it minimizes the contamination of
teats.
In free-stall and loose housing dairy barns, every management
technique which are available nowadays must be used.
As it insures that cows do not defecate in their stalls and increase
the level of contamination of teats.
3. Milking procedures(Pre and post milking teat disinfection and
using of post milking external teat sealant)
Many dairy producers have now incorporated premilking teat
disinfection into their mastitis control strategy. 845
 Mastitis Cont’d---
So that many different teat dips are being used.
The most commonly used premilking teat dip antiseptics
containing
 0.25% iodine
 0.1% - 0.25% iodophor
0.55% iodophor in 1.9% linear-dodecyl benzene sulfonic acid
Premilking and postmilking teat disinfection, in association with
good udder preparation, are significantly more effective in
prevention of environmental pathogen intramammary infection.
The use of postmilking external teat sealant has also significant
importance in preventation of bovine coliform mastitis.
Postmilking external teat sealant is a latex barrier teat dip which
form a physical seal between the teat and the environment.
So that teat sealant reduces the incidence of new coliform
intramammary infections during lactation.
A barrier teat dip containing 0.55% chlorhexidine was effective in
846
 Mastitis Cont’d---
reducing intramammary infection associated with both
environmental and contagious pathogens.
Nutrition
You know in animal nutrition course, vitamin E or selenium
deficiency decreases the innate immunity of animals and exposed
them to mastitis.
Because deficiency of vitamin E and/or selenium reduces
Neutrophil chemotaxis into the mammary gland
The intracellular killing capacity of neutrophils
Therefore, it is very important to ensure that vitamin E and
selenium intakes of dairy cows are adequate.
Preventation of bovine coliform mastitis in most intensive dairy
farms is best achieved
 By daily ingestion of 1000 IU of vitamin E and 3 mg selenium for
dry cows
By daily ingestion of 400 - 600 IU of vitamin E and 6 mg selenium
for lactating cows. 847
 Mastitis Cont’d---
Prevention of bovine coliform mastitis during dry period
Cows that are due to calve should be kept on grass or moved into a
clean area at least 2 weeks before calving.
Their udders and teats should be washed daily if necessary, and teat
dipping with a teat disinfectant should be begun 10 days before
calving.
This is particularly necessary for older cows and those that are
known to be easy milkers.
The teats of those cows that are ‘leakers' just before calving may
have to be sealed with adhesive tape or collodion to minimize the
chance of infection.
Recumbent cows
Cows that are recumbent and unable to stand (e.g. the downer cow)
should be well bedded on clean dry straw.
Their udders should be kept clean and dry, and the teats should be
dipped with a teat disinfectant.
Strict hygiene must be practiced when using teat siphons and teat848
 Mastitis Cont’d---
creams, and strict asepsis must be applied when you are doing teat
surgery.
Milking machine
Irregular vacuum fluctuations in the milking machine may induce
coliform mastitis in quarters exposed to a high level of
contamination.
The operation and sanitation of the milking machine, especially
those parts indirect contact with the teats, must therefore be
examined.
Vaccination using core lipopolysaccharide antigen vaccine
Vaccination of cows during the dry period and early lactation with
core lipopolysaccharide antigen vaccine is applied in many
countries of the world with developed intensive dairy production.
The core antigen (lipid A component) is uniform between bacterial
species possessing lipopolysaccharide and is immunogenic.
It provides one tool to reduce the incidence and severity of clinical
coliform mastitis. 849
 Mastitis Cont’d---
The two core LPS antigen vaccines that are a available nowadays
 Re mutant of Salmonella typhimurium
 Rc mutant of E. coli.
These vaccines are available in the USA.
But these vaccines are not widely used nowadays because of their
endotoxin content.
The endotoxin content causes
Reduction of leukocyte and blood segmented neutrophil
concentration( For instance, vaccination of late lactation and dry
cattle with the S. typhimurium Re mutant).
Reduction of milk production by 7% at the second and
third milkings after vaccination with the E. coli Rc mutant.
2.2. Bovine mastitis caused by environmental streptococci
It is environmental bovine mastitis in which the animals are
infected by the bacteria from the environment.
Etiology
So many species of Streptococcus and occasionally Enterococcus850
 Mastitis Cont’d---
may cause bovine environmental streptococcal mastitis.
But the most common are
Streptococcus uberis
Streptococcus dysgalactiae
S. dysgalactiae has characteristics of both a contagious and an
environmental pathogen.
However, some categorization schemes makes it in the contagious
category eventhough it is primarily an environmental pathogen.
Other uncommon environmental streptococci those are involved in
bovine mastitis include
Streptococcus equi var. zooepidermicus
Streptococcus viridians
 S. equinus formerly called S. bovis
Streptococcus pyogenes
Streptococcus pneumoniae
 Enterococcus spp.
. 851
 Mastitis Cont’d---
Epidemiology
Occurrence and prevalence of infection
The environmental mastitis caused by Streptococcus and
Enterococcus spp. is prevalent in country where intramammary
infections due to the contagious pathogens S. agalactiae and
S. aureus has been reduced or eradicated.
That is why the proportion of intramammary infections associated
with environmental streptococci has increased markedly in dairy
farms of the world.
Nowadays these organisms are a leading cause of both subclinical
and clinical mastitis in dairy cattle worldwide
Among them S. uberis is now a common cause of intramammary
infection.
It occurs mostly during the dry cow period with most clinical cases
occurring during the first months of lactation.
Environmental streptococcal intramammary infections are usually
short lived for about a month. 852
 Mastitis Cont’d---
However, small percentage of cases may become chronic.
There is a change in the epidemiology of bovine mastitis in dairy
farms of the world over the past decade.
It was the rise of environmental pathogens mainly causing clinical
mastitis relative to contagious pathogens.
Nowadays there is remarkable increases in both coliform and
environmental streptococci as causes of clinical mastitis in intensive
dairy farms of the world.
Source of infection
Fecal shedding is believed to play an important role in the
maintenance of S. uberis populations on dairy farms.
The number of S. uberis found in the dairy farms depend on the
type of bedding material used.
It was found that large numbers of S. uberis are found in straw
bedding and much lower numbers in sawdust and wood shavings.
S. dysgalactiae is also associated with summer mastitis.
It affects dry cows and heifers during the summer months. 853
 Mastitis Cont’d---
It has been isolated from the common cattle fly Hydrotaea irritans.
This fly may be involved in the establishment and maintenance of
bacterial contamination of teats.
Risk factors
The common risk factors for occurrence of environmental bovine
streptococci mastitis are
 1. Environmental risk factors
 2. Animal risk factors
 3. Pathogen risk factors
1.Environmental risk factors
The major risk factor for environmental streptococci infection is
exposure of the teat end to mastitis pathogens in the environment.
Transmission is predominantly from the environment.
You know exposure of uninfected teats to environmental
streptococci can occur during
Milking process
 Between milkings 854
 Mastitis Cont’d---
During the dry period and
Prior to parturition in first-lactation heifers
Other environmental risk factor is housing and management
practices.
Housing and management practices may contribute
 For contamination of bedding materials
 For exposure of teats to environmental streptococci.
Housing facilities that predispose to the accumulation of feces on
cows will increase the rate of exposure of the teat end to the
pathogens.
Type of bedding material used in dairy farms is risk factor.
Straw bedding appears to increase the risk of S. uberis mastitis than
saw dust and wood shaving.
Pastured cattle are generally at reduced risk for environmental
streptococcal mastitis when compared to cows in confinement
housing.
However, certain pasture conditions, such as areas under shade 855
 Mastitis Cont’d---
trees, poorly drained ground surfaces, ponds and muddy areas,
may result in a high rate of exposure to the pathogens.
S. dysgalactiae is commonly isolated from heifers and cows in the
dry period
It is one of the most prevalent pathogens isolated from cases of
summer mastitis.
The spread of S. dysgalactiae between cows within dairy herds
may occur directly or by way of the milking machine or
environment.
That is why S. dysgalactiae is considered as primarily as
environmental mastitis causing pathogen and secondarily as
contagious mastitis causing pathogen.
2. Animal risk factors
The risk of new infections is influenced by
The stage of lactation
Parity of the cow
The rate of new infection is highest during the 2 weeks following856
 Mastitis Cont’d---
drying off and the 2 weeks prior to calving.
The high rates of new infection following drying off may be
associated with
The lack of flushing action of milking
 The changes in the composition of the mammary secretion which
may enhance the growth of the pathogens
The lack of a keratin plug in the streak canal
The increase in susceptibility to infection just prior to parturition
may be associated with
The lack of milking when the gland is accumulating fluid
Loss of keratin plugs from streak canals
Immunosuppression of the periparturient period
The rate of infection is also higher in older cows than for either
heifers or cows in second lactation
Infection is highest during the summer months for both cows in
lactation and cows in the dry period.
857
 Mastitis Cont’d---
3. Pathogen risk factors
Several virulence factors of S. uberis and S. dysgalactiae have been
identified.
These virulence factors are important in the pathogenesis of
environmental mastitis.
The virulence factors of S. uberis and S. dysgalactiae is the
antiphagocytic factor of hyaluronic acid capsule .
Hyaluronic acid capsule allows S. uberis
To infect and multiply in the gland
 To adhere to and invade the mammary tissue
Bovine mammary macrophages are capable of phagocytosis of the
organism.
But certain strains of S.uberis are capable of resisting phagocytosis
by neutrophils.
Strains of S. dysgalactiae possesses several antiphagocytic factors
including
M-like protein 858
 Mastitis Cont’d---
Alpha-2-macroglobulin capsule
Fibronectin binding
 Hyaluronidase
 Fibrinolysin
Economic importance
The major economic losses associated with environmental
streptococcal mastitis are
Loss of production
Milk withholding time during treatment
Premature culling
increased labor costs of therapy and veterinary services.
Pathogenesis
During infection of dairy cows with S.uberis there is acute
inflammation of udder resulting in the accumulation of large
numbers of neutrophils in the secretory acini within 24 hours.
After 6 days of infection the neutrophil response is still evident.
But there is cellular infiltration, septal edema, extensive 859
 Mastitis Cont’d---
vacuolation of secretory cells and focal necrosis of alveoli.
At this stage, the organism is present free or phagocytosed in
macrophages in the alveolar lumina, adherent to damaged secretory
or ductular epithelium, in the sub epithelium and septal tissue and in
lymphatic vessels and lymph nodes.
Clinical findings
Approximately50% of environ mental streptococcal intramammary
infections cause clinical mastitis during lactation.
The clinical findings are usually limited to abnormal milk or
abnormal gland.
From the total infected herds of dairy farms
About 43% of cases the findings are limited to abnormal milk
About 49% involve abnormal milk and an enlarged (abnormal)
gland
Only 8% of cases are with systemic signs with clinical sign of
fever and anorexia (abnormal cow)
860
 Mastitis Cont’d---
Necropsy findings
Since there is no typical pathological changes that are visible in
gross pathology and in histopathology, it is not applicable.
Diagnosis
Diagnose can be done based on
Housing and management practice of the farm
Clinical signs
SCCs by CMT
Laboratory
Specimens can be milk, udder tissues and lymph nodes and
transported at 4 0C .
The bacteria can be isolated and identified in the laboratory.
Species can be differentiated with reasonable success using a
variety of biochemical tests, such as the API20 strep after
serological grouping using specific antisera called Lancefield
groups.
However, molecular biological techniques are needed for detail 861
 Mastitis Cont’d---
identification of the bacteria.
NB. All the environmental streptococci except S. dysgalactiae
hydrolyze aesculin on blood agar.
Differential diagnosis
Bovine mastitis caused by Streptococcus uberis in dry cows may be
sufficiently severe.
It resembles mastitis associated with A. pyogenes.
So that diagnoses dépends on cultural examination of the milk.
Treatment
Most isolates of S. uberis and S. dysgalactiae are susceptible to
penicillin, novobiocin, amoxicillin and cephapirin.
A high percentage (96%) are also susceptible to tetracycline.
But their susceptibility to amino glycosides is much lower.
Most cases of clinical mastitis associated with S. uberis and S.
dysgalactiae respond well to intramammary infusions of penicillin,
cephalosporins, cloxacillin, erythromycin and tetracyclines.
Rarely spontaneous cures can also occur. 862
 Mastitis Cont’d---
Clinical cases in lactating cows should be treated by at least two
intramammary infusions 12 hours apart.
Failure of treatment may be due to
Epithelial cell invasion of the bacteria
Movement of the bacteria into sub epithelial layers,
 These conditions possibly reduces the effectiveness of the
antimicrobial drugs
Extended therapy (for 5 or 8 days) with intramammary infusion
every 24 hours may increase the bacteriological cure rate.
Some of them are
Ceftiofur (125 mg)
Pirlimycin (50 mg)
Penethemate hydriodide
Dihydrostreptomycin sulphate
 Framycetin sulfate
Parenteral administration of antibiotic is necessary if there is
systemic infection( abnormal cow). 863
 Mastitis Cont’d---
Preventation and control measures
Preventation and control of mastitis due to environmental
streptococci is achieved by
Decreasing the exposure of pathogens to the teat end and
Increasing the resistance to intramammary infections.
One of the best method to control environmental streptococci
mastitis is not to bed on straw.
But this may not be a practical or economic recommendation
for some producers.
So that if straw bedding is used, a reduction in the teat end
exposure to S. uberis can result from frequent (daily) replacement
of bedding.
Reducing the exposure of the teat end to pathogens depends on
maintenance of a clean and dry environment.
Sand is the ideal bedding material because it has the lowest
number of coliform and environmental pathogens.
864
 Mastitis Cont’d---
Wet and damp areas in the back part of free stalls and tie stalls
promote exposure to environmental pathogens.
So that wet and damp areas of free and tie stalls must be clean and
dry.
The following practices must be performed in dairy farms to
prevent environmental streptococci mastitis.
Keep the dairy farms clean, dry and use ideal bedding materials
Predipping with a teat dip germicide like iodophor may reduce
environmental mastitis by as much as 50%.
 Application of an internal teat sealant of bismuth sub nitrate at dry-
off is effective in preventing infections associated with S. uberis
during the dry period.
 A long-acting intramammary infusion dry cow therapy containing
250 mg cephalonium administered after the last milking of lactation
reduced the incidence of new infections due to S. uberis.
865
 Mastitis Cont’d---
1.3. Bovine mastitis caused by Arcanobacterium pyogenes
It is one of environmental pathogens that cause environmental
bovine mastitis.
It causes two forms of severe bovine clinical mastitis namely
 Suppurative mastitis mostly in housed cattle and is referred to as
pyogenes mastitis which occurs in sporadic cases
 Summer mastitis which is clinically similar disease that occurs as
outbreaks in cattle caused by P. multocida and M.
haemolytica(formerly P. haemolytica)during the summer months in
Europe .
Etiology
It is caused by Arcanobacterium pyogenes (formerly Actinomyces
pyogenes and Corynebacterium pyogenes).
Epidemiology
Occurrence and prevalence of infection
There is a much higher incidence of suppurative ‘'summer
mastitis ‘’during the summer months when non lactating females
866
 Mastitis Cont’d---
are left at pasture and not kept under close observation.
Bovine mastitis associated with A. pyogenes occurs sporadically
and is most common in dry cows or pregnant heifers.
Rarely lactating cows may also be affected.
Source of infection and mode of transmission
The portal of infection is unknown although it is presumed to be via
the streak canal.
The method of spread is uncertain in sporadic cases but insects,
especially biting ones such as Hydrotoea irritans appear to play an
important role in outbreaks of 'summer mastitis
That is why the prevalence of the disease is related to the peaks of
the fly populations and the prevailing climate which is wet and
warm(summer).
Risk factors
The common risk factors for occurrence of the disease are
 1. Environmental risk factors
 2. Stage of lactation 867
 Mastitis Cont’d---
3. Occurrence of concurrent diseases
1. Environmental risk factor
The incidence is much higher in wet summers and on heavily
wooded and low-lying farms when the fly population is high
Heavy fly populations are a common accompaniment of an
outbreak.
The infection rate of A. pyogenes in udders is much less in housed
cattle than in the same cattle at pasture.
It has been suggested that some triggering mechanism is needed
before contamination of the teat and invasion and infection of the
gland can occur.
2. Stage of lactation
Dairy breeds are the predominant target, mostly in dry cows or
pregnant heifers.
Rarely the disease can affect lactating cows .
3. Occurrence of concurrent diseases
Lactating cows usually acquire the infection after injury or the 868
 Mastitis Cont’d---
development of black spot on the teat.
Severity of the disease is determined by the presence of anaerobes
like F. necrophorum.
Outbreaks of pyogenes mastitis are also recorded in association
with outbreaks of foot-and mouth disease and herpes mammilitis.
Because these viruses cause damage to the teats.
Economic importance
Summer mastitis caused by P. multocida and M. haemolytica is a
serious disease if adequate treatment is not done it cause
Death with mortality rate about 50%
Totally destruction of the affected quarters of surviving cows.
However, in pyogenes mastitis which is caused by A. pyogenes
the mortality rate is much less.
But it causes total destroy of the affected quarter.
So that the loss of the quarter means that the cow is culled.
Pathogenesis
Even if the organism is applied to the teat skin at the end of the 869
 Mastitis Cont’d---
teat, infection of the quarter does not occur unless
The teat end is injured and
 Anaerobic bacteria are involved in the infection
It is suggested that the infection is carried from udder to udder by
flies.
The massive invasion of the mammary tissue occurs via the teat
canal that is damaged.
The greater part of the gland is affected at the first attack, causing a
severe systemic reaction and loss of function of the entire quarter.
Clinical findings
Mastitis associated with A. pyogenes is usually peracute with a
severe systemic reaction including
Fever (40 - 41°C)
Rapid heart rate
Complete anorexia
Severe depression and weakness
Abortion may occur during this stage 870
 Mastitis Cont’d---
In almost all cases only one quarter is affected most commonly the
front one
The teat is swollen and inflamed and the quarter is very hard,
swollen and sore
The secretion is watery with clots early and later purulent with a
typical putrid odor.
Affected cows usually carry a large fly population.
If the cow survives severe toxemia develops
The quarter becomes extremely indurated and abscesses develop
Later rupturing through the floor of the udder commonly at the
base of the teat.
Sloughing of the affected quarters
The function of the quarter is permanently lost and cows that have
calved recently may go completely dry.
Severe thelitis with extreme thickening and obstruction of the teat
is the common sequel of mastitis caused by A. pyogenes.
Lameness in the hind limb on the affected side occurs in some 871
 Mastitis Cont’d---
cases, and the limb joints may be swollen.
Necropsy findings
Details of the pathology of the disease are not available.
Diagnosis
Diagnose can be made based on history, epidemiology, clinical and
laboratory.
Confirmatory diagnose is done by isolation and identification of
the bacteria.
Milk, pus from abscess can be used as specimens
Freezing of milk samples reduces the number of samples giving a
positive cultural result.
Differential diagnosis
Mastitis caused by A. pyogenes must be differentiated from many
mastitis that have almost similar clinical features.
But it is easy to differentiated this mastitis because it has
Seasonal incidence in some areas
 Acute inflammation of one quarter mostly the fore quarter. 872
 Mastitis Cont’d---
Suppurative nature of mastitis
Abscesses formation and
Severe systemic reaction
Treatment
Summer mastitis caused by A. pyogenes normally responds poorly
to treatment.
Most of the affected quarter is typically lost for milk production.
Failure of therapy is due to the extensive purulent processes in the
udder but not due to antimicrobial resistance.
Experimentally it is known that bacterial isolates from cases of
summer mastitis caused by A. pyogenes are susceptible to penicillin
G and to other beta -lactam antimicrobials “in vitro” but not “in
vivo”.
Because penicillin G has limited distribution throughout the
inflamed udder due to purulent pus formation.
In peracute cases administration of the following antimicrobial
drugs in combination with other methods give a better result. 873
 Mastitis Cont’d---
 Parenteral administration of sodium sulphadimidine or one of the
tetracycline groups in combination with intramammary infusion of
broad spectrum antibiotics accompanied by repeated stripping of
the quarter.
 The best result is obtained by permanently drying off a quarter
and infusion of 120 ml of 5 % povidone-iodine solution (0 .5%
iodine) after complete milk-out accompanied by administration of
flunixin meglumine (1 mg/kg BW, intravenously).
In the above methods of treatment clearing of proteinaceous debris
from the affected quarter is essential to increase the efficiency of
treatment.
Clearing of proteinaceous derbies from the udder may be aided by
the intramammary application of proteolytic enzymes( artificial
trypsin).
NB. In summer mastitis caused by A. pyogenes, even with intensive
therapy, at least 80% of quarters are rendered useless and many of
those that respond are greatly reduced in productivity. 874
 Mastitis Cont’d---
Preventation and control measures
The most favored techniques used to control and prevent summer
mastitis caused by A. pyogenes are
 Intramammary infusion a dry cow with Cloxacillin 500 mg and
ampicillin 250 mg in a long-acting base at 3 -week intervals during
the dry period.
 An alternative intramammary infusion procedure is to use
cephalonium at 4-weekly intervals.
Repeated spraying of the udder, for example automatically at
watering points, with a contact insecticide is commonly carried out
during the fly season and is believed to be effective.
In particular, cows with purulent material draining from an affected
quarter need to be isolated from other cattle.
Early treatment of teat lesions is very important to limit bacterial
colonization and to limit possibly transported by flies.
Careful daily examination of dry cows during the summer.
Careful daily examination of dry cows during the summer may 875
 Mastitis Cont’d---
enable
To identify the affected quarters
 To identify cows to be isolated
 To treat the affected quarters at an early stage
III. Mastitis of cattle associated with teat skin opportunistic
pathogens
3.1. Bovine mastitis caused by coagulase negative staphylococci
They are among the most common bacteria found
in milk.
Coagulase negative staphylococcal bovine mastitis are common
especially in herds in which the major pathogens have been
adequately controlled.
There is considerable debate about the significance of these
pathogens for the mammary gland and for cow productivity.
For this reason, these pathogens are classified as minor pathogens
of udder.
876
 Mastitis Cont’d---
Etiology
Coagulase-negative staphylococci that can cause bovine mastitis
include
Staphylococcus epidermidis
 S. chromogenes
 S. simulans
Staphylococcus warneri
These bacteria are normal teat skin flora.
However, there are coagulase – negative staphylococci that come
from uncertain sites which can cause bovine mastitis.
Some of the common are
 Staphylococcus xylosus
Staphylococcus sciuri
Epidemiology
Coagulase-negative staphylococci are teat skin opportunistic
pathogens.
They cause mastitis by ascending infection via the streak canal.877
 Mastitis Cont’d---
Coagulase –negative staphylococci appear to have a protective
effect against colonization of the teat duct and teat skin by
S. aureus and
Other major pathogens with the exception of E. coli and the
environmental streptococci.
These bacteria can cause mastitis in dairy farms where the major
mastitis pathogens are controlled.
These bacteria mostly affects heifers in peripartum and post
parturition.
Almost 50% of heifer quarters are infected before parturition with
coagulase –negative staphylococci.
That is why Coagulase-negative staphylococci are the most
commonly isolated bacteria from milk samples of heifers with
mastitis.
But these infections are usually eliminated spontaneously or with
antimicrobial therapy during early lactation
878
 Mastitis Cont’d---
Clinical findings
Coagulase-negative staphylococci are usually associated with mild
clinical disease which is characterized by
Abnormal secretion only
Occasionally abnormal gland(swelling)
They are commonly isolated from mild clinical cases of mastitis
and subclinical infections.
They usually induce a moderate increase in SCCs
Necropsy findings
They cause microscopic lesions but gross necropsy reports are
lacking.
Diagnosis
Diagnose can be done on the basis of
History
Clinical signs
SCCs
Laboratory 879
 Mastitis Cont’d---
Intramammary infections by minor pathogens such as coagulase-
negative staphylococci result in a higher than the normal SCCs.
So that CMT can also help to diagnose the disease.
This helps and there by increase the resistance of the colonized
quarter from invasion by a major pathogen.
Laboratory method of diagnosis is based on isolation and
identification of the bacteria.
Differential diagnosis
This type of mastitis must be differentiated from other mastitis that
cause mild form of mastitis.
Treatment
Spontaneous cure is common.
Coagulase negative staphylococci, including are very susceptible to
penicillin, ampicillin, amoxicillin, clavulanic acid, cephapirin,
erythromycin, gentamycin.
The efficacy of the above antimicrobial drugs can be potentiated
with sulfonamides and tetracyclines. 880
 Mastitis Cont’d---
The use of a combination of novobiocin and penicillin and
cloxacillin as dry cow therapy for coagulase-negative staphylococci
gave cure rates of over 90%
All the above antimicrobial drugs are administered intramammary.
Preventation and control measures
Implementation of a mastitis control program will be very effective
in decreasing intra mammary infection due to coagulase negative
staphylococci.
4. Mastitis of cattle associated with less common pathogens
The less common pathogens that cause mastitis in cattle
Causative agent Reasons of Clinical feature of mastitis
occurrence
Pseudomonas Intramammary It occurs sporadically
aeruginosa and other infusion with Acute mastitis with systemic reaction
spp. of Pseudomonas contaminated and swelling of the udder and
material. the appearance of clotted, discolored
Milk which is accompanied with 17%
881
mortality.
 Mastitis Cont’d---
The less common pathogens that cause mastitis in cattle
Causative agent Reasons of Clinical feature of mastitis
occurrence
It may cause subacute and chronic
mastitis
Pasteurella species Occurrences in Summer mastitis
mainly P. multocida summer sporadically Severe with fever, profound toxemic
and M. haemolytica It has/ has no relation shock, weak pulse, tachycardia, and
with and with out A. with mastitis caused recumbency.
pyogenes. by A. pyogenes The affected quarter is very swollen
It has relation with The milk is watery, red-tinged and
fly population contains flakes.
All four quarters may be affected.
Subsequent fibrosis and atrophy of
udder is common.
Newborn calves allowed to suck
colostrum from affected cows
may die ofpasteurellosis.

882
 Mastitis Cont’d---
The less common pathogens that cause mastitis in cattle
Causative agent Reasons of occurrence Clinical feature of mastitis

Nocardia species Accidental introduction Systemic reaction with high fever,

mainly Nocardia of the causative bacteria depression and anorexia.
asteroides into udders when An acute or subacute inflammation
Rarely infusions. is more usual.
N.brasiliensis Confinement of dairy Subacute mastitis accompanied by
andNocardia cattle in muddy pens. extensive granulomatous lesions in
farcinica Because the bacteria is the udder.
common soil contaminant Fibrosis of the gland and the
and probably gains appearance of clots in a grayish,
entrance to the udder viscid secretion that also contains
when udder washing is small, white particles.
ineffective or udder The fibrosis may be diffuse but is
infusion is not carried usually in the form of discrete
out aseptically. masses 2-5 cm in diameter.
Badly affected glands become
grossly enlarged, and may rupture
883
develop sinus tracts to the exterior.
5. Miscellaneous causes of bovine mastitis
1. Some common miscellaneous bovine mastitis caused by
bacteria
Infectious agent Reasons of occurrence Clinical sign

Bacillus species Contamination associated Peracute to acute hemorrhagic mastitis

Bacillus cereus with teat injuries or affecting one or more quarters.
and Bacillus surgery. Fever upto 40 - 41 0C and sever toxemia
subtilis Mastitis may also occur in Severe swelling and pain
cows at the time of calving Secretions are red-tinged and serous in
and is associated with the consistency.
feeding of brewers' grains Affected cows are weak and quickly become
in which the spores of B. recumbent
cereus are present. Death may occur in 24 - 36 hours.
Gangrene may occur and in cows that
survive, portions of affected gland will
slough out and a chronic relapsing mastitis
will persist.
Campylobacterct Not well studied. It has mostly subclinical form with high
er jejuni It has very rarely SCCs
occurrence.
884
 Mastitis Cont’d---
Some common miscellaneous bovine mastitis caused by bacteria
Causative agent Reasons of occurrence Clinical feature of mastitis
Clostridium Contamination of the teat High fever, swelling and
perfringens with soil that has spore. superficial hyperemia of the
Type A Trauma and other factors affected quarter.
create anaerobic condition It is followed later by gangrene,
in the udder and the Enlargement of the
bacteria release toxin supramammary lymph nodes
A thin brown secretion
containing gas
Subcutaneous emphysema
Fusobacterium The portal of infection is No systemic reaction
necrophorum unknown, Affected quarters have a
although it is presumed to be viscid, clotty, stringy secretion
via the streak canal from the There is little fibrosis.
soil since it is anaerobe.
Complicates summer mastitis
with other anaerobes

885
 Mastitis Cont’d---
Some common miscellaneous bovine mastitis caused by bacteria
Causative agent Reasons of occurrence Clinical feature of mastitis
Histophilus somni Contamination from the Cause an acute form with high fever
(formerly Haemophilus environment through teat and bloodstained milk.
somnus) canal. It may cause mild upto chronic
Infection of the mammary mastitis.
gland from systemic Mastitis at the end may have a
infection. gangrenous form
Listeria monocytogenes Infection through teat canal It cause subclinical mastitis and
from the environment. abnormal milk is rare.
Infection of the mammary But the affected quarter loses
gland from systemic productivity
infection SCCs usually greater than
107cells/ml of milk.
Pathogenic During bovine Udder losses its configuration
Mycobacterium spp. tuberculosis infection Affected quarter/s hard in palpation
M. bovis because of granulomatous lesion
Milk contains pus to a watery
fluid containing flakes
886
 Mastitis Cont’d---
Some common miscellaneous bovine mastitis caused by bacteria
Causative agent Reasons of Clinical feature of mastitis
occurrence
Saprophytic
Mycobacterium spp
1. M. lacticola Intramammary infusion Tremendous hypertrophy of the quarter
of therapeutic agents in Appearance of clots in discolored milk
oils but not in watery There is no systemic reaction.
suspension. Affected animals do not show
sensitivity to avian or mammalian
tuberculin.

2.Mycobacterium Intramammary infusion Infected quarters are

fortuitum, of therapeutic agents in seriously damaged and do not respond
Mycobacterium oils but not in watery to treatment, and affected cows die or
smegmatis and suspension. are salvaged.
Mycobacterium
chelonei.

887
 Mastitis Cont’d---
Some common miscellaneous bovine mastitis caused by bacteria
Animals show positive reactions
to mammalian and avian
tuberculosis and some sensitivity
to johnin.
The mammary secretion of
affected quarters varies from pus
NB. It is better to use watery to a watery fluid containing flakes
suspension of intramammary
infusion to prophylaxis the
disease.

Serratia species Using of contaminated Mild chronic mastitis in which

Serratia marcescens sawdust as bedding and swelling of the quarters
and inadequate cleaning of the with clots in the milk appear
Serratia liquefaciens teats before milking periodically.

888
 Mastitis Cont’d---
Some common miscellaneous bovine mastitis caused by fungi and
yeast
There are some species of fungi that causes bovine mastitis
Some of them are
Trichosporon spp.
Cryptococcus neoformans
Many other yeasts may cause bovine mastitis
Some of them are
Candida spp
Saccharomyces spp
Pichia spp
Torulopsis spp
 Aspergillus fumigatus
889
Some common miscellaneous infectious agents(viruses)of the
external surface of bovine udder and teat
Infectious agent Disease Clinical sign
Bovine parapoxvirus Pseudocowpox Starts as a small inflammatory
(Poxviridae) papule usually on teats that
develops into a dark red scab. The
central area desquamates and leave
a ring or horse shoe shaped scab.
Bovine herpesvirus 2 Bovine ulcerative The lesions are usually on teats but
mamillitis can spread to the udder. Lesion
starts as local thickening of the skin
followed by severe oedema and
erythema.
Bovine papillomavirus Bovine papillomas Cutaneous fibropapilloma that can
(Papovaviridae) Type 1 and 2 be large and cauliflower. Usually
occurs on head and neck but can be
present on the skin of the udder.

890
 Mastitis Cont’d---
Some common miscellaneous infectious agents(viruses)of the
external surface of bovine udder and teat
Infectious agent Disease Clinical sign
Type 5 Teat fibropapilloma with a rice –
grain appearance being flat and
white
Type 6 Teat papilloma or ‘ frond warts’
that are thin and pedunculated
Bovine orthopoxvirus Cowpox Forms pustules on teats and
(Poxviridae) (Vaccinia virus ruptures.
infection) The exudate forms a scab covering
an ulcerative area.
Apthovirus(Picornaviridae Foot and mouth Vesicles on the teat of milking cow
) disease
Vesiculovirus Vesicular Vesicles may occur on the teats of
(Rhabdoviridae) stomatitis milking cow .
Orbivirus (Reoviridae) Blue tongue Lesions on the teats of milking
cows 891
 Mastitis Cont’d---
3. Mastitis in domestic animals other than cattle
Mastitis has the greatest prevalence and economic importance in
dairy cows.
But it can be significant in other domestic animals particularly in
sheep, goats, pigs and equines.
3.1. Mastitis in ewes
Most causes of clinical mastitis in sheep is caused by
 80 - 90% of clinical mastitis is caused by Staphylococcus aureus
The remainder10 – 20 %) is due to
 Streptococcus agalactiae
M. haemolytica, with rare occurrences of Escherichia coli and
Histophilus somni(formerly Histophilus ovis) may cause mastitis in
ewes.
However coagulase negative staphylococci are a major cause of
subclinical mastitis in sheep.
A small percentage of cases are associated with
Clostridium perfringens A 892
 Mastitis Cont’d---
 Pseudomonas spp.
Corynebacterium pseudotuberculosis
 Acholeplasma oculi may cause mastitis and agalactia in ewes
Epidemiology
Occurrence
Clinical mastitis in ewes occurs about equally immediately after
weaning or close to parturition.
In pastured ewes the prevalence is low if it occurrences it is usually
due to either
M. haemolytica or Staphylococcus spp.
Clinical mastitis in pastured ewes averages only about 2 % per year.
But mastitis causes upto 10% of all ewe deaths.
Staphylococcal mastitis of ewes is the most prevalent mastitis in
ewes and is caused by S. aureus.
The mortality rate varies between 25 % and 50%.
The affected quarters in surviving ewes are usually destroyed.
893
 Mastitis Cont’d---
Clinical mastitis of ewes because of S. aureus is probably spread
from infected bedding grounds.
The infection gaining entry through teat injuries caused by sucking
lambs.
Other coagulase negative staphylococci like S. epidermidis, S.
xylose, S. simulans and S. chromogenes can cause subclinical
mastitis in ewes.
Streptococcal mastitis of ewes occurs naturally in sheep used as
milking animals.
The infection originates from an infected udder and is transmitted
to the teat skin of other females by
Milking machine liners
Milkers' hands
Wash cloths
Any other material that can act as an inert carrier
The common occasional causes of mastitis in ewes are
Streptococcus dysgalactiae 894
 Mastitis Cont’d---
Streptococcus uberis
Mannheimia mastitis of ewes causes peracute and gangrenous
form of mastitis.
It is mostly associated with Mannheimia spp. commonly with M.
haemolytica.
It is mastitis of ewes which is common in pasture .
S. aureus, Arcanobacterium pyogenes and streptococci are often
present as secondary invaders in manneheimma mastitis of ewes.
Mannheimia mastitis is most common in ewes suckling large lambs
2 - 3 months old.
Infection is thought to occur through injuries to teats, perhaps
caused by over vigorous sucking by big lambs.
Pathogenesis
The mechanisms of pathogenesis are similar to those for bovine
mastitis.
895
 Mastitis Cont’d---
Clinical findings
The main clinical findings those are observed in ewes in mastitis caused by
different bacteria are as follows
Types of mastitis in Main clinical signs observed
ewes
1. Staphylococcal mastitis It has both peracute and gangrenous infections.
caused by S. aureus The ewe is usually recumbent and profoundly toxic
The affected gland and the surrounding area of belly
wall are blue green in color and cold to the touch.
A few drops of clear, bloodstained liquid in the milk
2. Mannheimia mastitis An acute systemic disturbance, with a high fever (40 -
caused by M. haemolytica. 42°C) anorexia, dyspnea and profound toxemia
Acute swelling of the gland and severe lameness on
the affected side.
Usually only one side is affected
The udder is at first hot, swollen and painful and the
milk watery.
Within 24 hours the quarter is discolored blue-black
and cold. 896
 Mastitis Cont’d---
The main clinical findings those are observed in ewes in mastitis caused by
different bacteria are as follows
Types of mastitis in Main clinical signs observed
ewes
Within 24 hours the quarter is discolored blue-black
and cold.
The secretion is watery and red and contains clots.
The secretion dries up entirely and the animal either
dies
of toxemia in 3 - 7 days or survives with sloughing of
a gangrenous portion of the udder, followed by the
development of abscesses and the continual draining
of pus.
Cases of pneumonia due to the same organism
may occur in lambs
3. Clostridial mastitis It occurs rarely and highly fatal
caused by Clostridium It causes acute mastitis in ewes
perfringens A Clinical signs of infection are principally hemolytic
and are characterized by 897
Haemoglobinuria
 Mastitis Cont’d---
The main clinical findings those are observed in ewes in mastitis caused by
different bacteria are as follows
Types of mastitis in ewes Main clinical signs observed
Anemia
Jaundice
Fever
Anorexia and recumbency.
The affected quarter is swollen, painful and hot
The affected udder contains watery, brown and
flocculent secretion
Caseous lymphadenitis and Suppurative lesions are commonly found in ovine
mastitis caused by mammary g lands.
Corynebacterium But they usually involve only the supramammary lymph
pseudotuberculosis nodes and are not true mastitis.
The function of the mammary gland may be lost when
the infection has spread from the lymph node to
mammary tissue.
Pseudomonal mastitis Naturally occurring pseudomonal mastitis
caused by Pseudomonas in ewes is likely to be gangrenous and
aerogenes lethal and accompanied by severe lameness
898
 Mastitis Cont’d---
The main clinical findings those are observed in ewes in mastitis
caused by different bacteria are as follows
Types of mastitis in ewes Main clinical signs observed
in the hind limb on the affected side. Infected
intramammary infusions or
milking machine malfunction are the usual
means of introducing the infection

Necropsy findings
The gross appearance of the affected glands varies with the agent
involved and the duration of the process.
In general, the swollen, hemorrhagic, and/or gangrenous nature of
fatal acute ovine mastitis is glaringly obvious.
A purulent exudate is sometimes present, especially in the case of
chronic C. pseudotuberculosis infection.
899
 Mastitis Cont’d---
Diagnosis
Diagnose of mastitis in ewes can be done based on
History
Clinical signs
SCCs
Laboratory diagnose
Determination of the SCC is useful for the prediction of mammary
gland infection
The SCC in normal ewes' milk may range from 500 000-1 000 000
cells/ml in sheep
All mastitis-positive samples had somatic cells in excess of 2 000
000 cells/ml .
It is suggested that the threshold level for subclinical mastitis in
ewes should be close to 1 500 000 cells/ml.
Confirmatory diagnose can be done by isolation and identification
of the causative agents.
900
 Mastitis Cont’d---
Samples for confirmation of diagnosis are chilled mammary gland
for aerobic culture and anaerobic culture if Clostridium spp. is
suspected.
Differential diagnosis
Mannheimia mastitis is peracute and must be differentiated from
mastitis associated with S. aureus.
Suppurative mastitis associated with C. pseudotuberculosis is
chronic in type with out systemic signs.
It must be differentiated from Actinobacillus lignieresii which
cause a similar disease in ewes.
Differentiation of clostridial mastitis is also necessary.
All these are differentiated in laboratory by isolation and
identification of the causative agents.
Treatment
Broad-spectrum parenteral and intramammary antimicrobial agents
are effective.
Systemic treatment is necessary if there is systemic reaction. 901
 Mastitis Cont’d---
Ewes probably require smaller doses of intramammary infusions
than cows
So that it is customary to use ordinary cow-type mammary infusion
treatments.
The treatment of ewes with peracute gangrenous mastitis is as
unsatisfactory in terms of results as in cows.
Preventation and control measures
Removal of sources of infection by culling of some ewes in sheep
flocks necessary to control the disease.
However, even rigid culling usually fails to completely eradicate
the disease.
Dry period treatment with intramammary infusions of cephapirin
benzathine (1 infusion of both halves) is being greatly reduces the
subsequent new infection rate.
It is possible that vaccination could be an effective method of
control for staphylococcal mastitis in developed countries.
Prophylactic infusion of each half of the udder within a few days902of
 Mastitis Cont’d---
weaning, using half of a tube of dry cow treatment of penicillin and
streptomycin reduces mastitis in ewes.
The frequent changing of pasture areas and culling of affected ewes
should also help to control the spread of infection.
In developed countries for mannheimia mastitis in ewes use
polyvalent hyperimmune serum and a formalized vaccine have been
shown to be of value in prophylaxis.
3.2. Mastitis in shoots
Goats are much less frequently affected by contagious mastitis than
cattle and ewes.
But mastitis is an important sign in many of the infectious diseases
associated with Mycoplasma agalactiae and Mycoplasma mycoides
var. mycoides.
The most common mastitis in shoots are
1. Staphylococcal mastitis
2. Streptococcal mastitis
903
 Mastitis Cont’d---
3. Pseudomonas mastitis
4. Summer mastitis
5. Mastitis caused by other infectious agents
1. Staphylococcal mastitis
This is the most common mastitis in goats caused by S. aureus.
As in cattle, some staphylococci in goats' milk produce enterotoxins
and the toxic shock syndrome toxin(TSST) and are likely to cause
food poisoning in humans.
2. Streptococcal mastitis
Goats are uniformly susceptible to as cattle to Streptococcus
agalactiae.
In flocks of milking goats the infection is passed from infected
quarters to others by means of
 Milkers’ hands
 Teat cups of milking machines
 Washing cloths used to disinfect the udder before milking
904
 Mastitis Cont’d---
Streptococcus zooepidermicus causes chronic suppurative mastitis
in shoots.
3. Pseudomonas mastitis of shoots
Mastitis in shoots which is caused by Pseudomonas mastitis in
goats is acute, with extensive necrosis and fatal septicemia.
4. Summer mastitis of shoots
Summer mastitis in shoots is mainly associated with A. pyogenes.
It causes udder lesions typical of acute suppurative mastitis.
5. Mastitis in shoots caused by other infectious agents
Mastitis in goats is associated with an organism tentatively
identified as M. haemolytica and Yersinia pseudotuberculosis cause
mastitis in an aborting shoot.
The infection would have had zoonotic implications.
Granulomatous lesions in the mammary glands and in internal
organs have been observed in goats experimentally infected with
Cryptococcus neoformans.
905
 Mastitis Cont’d---
Clinical findings
Clinical mastitis in goats has similar to that of in cattle depending
on the form of mastitis.
Mastitis in shoots may show similar clinical features as cattle
Mastitis in goats has the following forms
Peracute
Acute
Subacute
Chronic
Gangrenous form
Diagnosis
Diagnose can be made based on
History
Clinical
SCCs using CMT
Laboratory
906
A physiological threshold of SCCs of 500 000 cells/ml has
 Mastitis Cont’d---
been suggested.
While SCCs of more than 1 000 000 cells/ml can b e regarded as
positive for mastitis.
The definitive diagnose is made based on isolation and
identification of the causative agents.
Latex agglutination tests are available for the identification of the
enterotoxins produced by S. aureus in the mastitic milk.
Treatment
Treatment of shoots is advisable either intramammary and/ or
parenterally depending forms of mastitis.
Treatment can be given either in dry period or lactation period
based on the result of laboratory findings.
If the cow dose rate is used retention of the antibiotic in the udder
will be prolonged and the holding period will need to be increased.
Intravenous administration of anti-inflammatory and antipyretic
agent like flunixin meglumine (two doses at 2.2 mg/kg BW hours)
or dexamethasone (0.44 mg/kg BW once) increase efficacy of 907
 Mastitis Cont’d---
treatment.
3.3. Mastitis of mares
Mastitis in mares occurs rarely.
The most common causative agent of mastitis in mares are
Corynebacterium pseudotuberculosis
Pseudomonas aeruginosa
Streptococcus zooepidermicus
Streptococcus equi
Streptococcus pyogenes
Staphylococcus aureus
Escherichia coli
Klebsiella spp.
Neisseria spp.
Beta hemolytic streptococci have been found in the milk of many
normal, just-foaled mares.
Clinical cases occur at any time during the lactation and many
occur in non lactating mares. 908
 Mastitis Cont’d---
Clinical findings
In streptococcal mastitis there may be severe local pain and
moderate systemic signs.
In most cases both halves are affected.
Gangrenous mastitis similar to that in cows may occur
occasionally.
Severe cases, sometimes accompanied by
Fever
Depression
Anorexia
Swelling
Pain and heat in the affected half
 Ventral edema and clots in the milk
The mare is lame in the leg on the affected side.
Gangrene and sloughing of the ventral floor of a gland may occur.
Treatment
909
Because of the high frequency of Gram-negative bacteria as
 Mastitis Cont’d---
causative agents in mares, treatment should include a broad-
spectrum antibiotic in an intramammary infusion plus intensive
parenteral antibacterial treatment.
Trimethoprim sulfonamide preparations are generally satisfactory.
Hot packs in chronic mastitis and frequent milking are also
recommended.
V. Complication of mastitis
If different forms of mastitis are not treated appropriately on time,
it results in complication.
There are two forms of complication of mastitis.
These are
 1. Induration of udder
 2. Gangrene of udder
910
 Mastitis Cont’d---
1. Induration of udder
It is the growth of intra alveolar connective tissues which is
accompanied with atrophy of its parenchyma.
Induration of udder occurs during mastitis when inflammatory
process destructs the parenchyma of udder( glandular alveolar
epithelium ) is completely replaced by connective tissues.
Induration of udder is irreversible.
That is why treatment does not give a good result.
So that culling of the animals out of production is necessary.
Use the animals for meat after fattening.
2. Gangrene of udder
Gangrene of udder occurs when mastitis or wound of udder is
complicated with anaerobic bacteria.
Clinical findings
It grows very fast and accompanied by high body temperature.
It occurs in septicemic or pyemia forms.
911
 Mastitis Cont’d---
In udder develops ischemia, edema and coldness of the affected
quarter.
Skin of the udder will have black – red colour with blue spots.
Affects quarter develops gangrene from which exudate comes
which has bad smelling.
On the base of affected quarter there is the formation of line which
has red – blue colour.
During milking there is small amount red – gray secretion in the
milk with bad smell.
Prognosis is bad.
Treatment
Isolation of the affected animals in calm animals.
To stimulate the condition of the animals, it is important to
administer IV 40% solution of glucose, 10% cacl2 , Caffeine and
IM antibiotic.
Intramammary infusion of solution of ligula, oxygen, hydrogen
912
peroxide and solution of potassium permanganate.
 Mastitis Cont’d---
After 15 minutes all the solutions are removed from udder through
intramammary catheter.
Then intramammary antibiotic infusion is necessary.
It is important to open the lesion of gangrene surgically and to
wash the wound with hydrogen peroxide, put powder of iodophor
or streptocida and potassium permanganate (3: 1) after every 4
hours lesion of gangrene.
Finally if treatment does not give good result, cull the animal.

913
914
In this portion we give attention on only
1.1. Rabies

915
 Rabies is a Latin word for madness.
Rabies can be defined as an invariably fatal neurologic disease
affecting most warm-blooded animals.
The disease called rabies encephalomyelitis.
It is manifested by
 Motor irritation with signs of mania and an attack complex
Inability to swallow and
Progressive paralysis beginning in the pelvic limbs.
Etiology
Rabies is caused by a rhabdovirus.
Rhabdovirus is bullet shaped RNA virus.
Rabies virus belongs to
Order of Mononegavirales
 Family of Rhabdoviridae
 Genus Lyssavirus
916
 Rabies cont’d---
 Genus Lyssavirus
The genus is composed of at least seven genotypes.
There are two types of rabies virus.
They are
 Fixed viruses refer to laboratory strains of low virulence and
 Street viruses refer to isolates of rabies virus from naturally
infected field cases.
It is now known that there are several strains of rabies virus.
They are adapted to particular host species but remain infective for
any warm-blooded mammal.
The virus is truly neurotropic and one of the larger viruses.
The virus is relatively fragile and susceptible to most standard
disinfectants and dies in dried saliva in a few hours.
Epidemiology
Occurrence
Rabies occurs in all warm-blooded animals.
Rabies is common in most countries of the world. 917
 Rabies cont’d---
It may be absent in some insular countries that exclude it by rigid
quarantine measures or prohibition of the entry of dogs.
All warm-blooded animals with the possible exception of opossums
are susceptible to rabies.
There is no variation in susceptibility with age.
However, variation in susceptibility between species is noticeable.
Foxes, cotton rats and coyotes are extremely susceptible
Cattle, rabbits and cats are highly susceptible
 Dogs, sheep and goats are moderately susceptible
Opossums little if at all.
Rabies viruses are well fitted to vectors belonging to the orders of
Carnivora (flesh-eating mammals including skunks)
 Chiroptera (bats)
So that bats and carnivora are the major reservoir of the disease.
Reservoirs of rabies virus are different in different continents of
the world.
 Raccoons and skunks are major reservoir of rabies in North 918
 Rabies cont’d---
 America.
Vampire bat in Latin America
Wild carnivores including mongoose in Africa
Method of transmission
Source of infection for rabies is almost always an infected animal.
Method of spread is almost always by the bite of an infected animal.
Although contamination of skin wounds by fresh saliva may result
in infection.
Rarely contamination of skin wound by fresh saliva as well as
ingestion of virus (if the dose is large enough) can spread rabies.
Not all bites from rabid animals result in infection.
Because the virus is not always present in the saliva and may not
gain entrance to the wound.
The virus may appear in the milk of affected animals but spread by
this means is unlikely as infection.
The natural occurrence of rabies in animals in caves inhabited by
infected insectivorous bats inhalation as a route of infection came919
 Rabies cont’d---
under suspicion.
It is now accepted that inter bat spread and spread from bats to other
species is principally by bites.
But that infection by inhalation also occurs.
Traditionally the dog and, to a minor extent cat are considered as the
main source infection for animals.
Native fauna including foxes, skunks, wolves, coyotes, vampire,
insectivorous, fruit-eating bats, raccoons, mongoose and squirrels
provide the major source of infection in countries where domestic
carnivora are well controlled.
Bats represent a serious threat of spread of rabies.
Because of their migratory habits.
Most spread is within the species
But the threat to humans and animal species by bats cannot be
completely disregarded.
Spread of the disease is often seasonal with the highest incidence in
the late spring around April – beginning of June) and at the end of
920
 Rabies cont’d---
winter and beginning of autumn.
This is because of it is the time of
Large scale movements of wild animals
Beginning of mating of domestic and wild carnivora
Zoonotic implications
The prime importance of rabies is its transmissibility to humans,
with veterinarians being at special risk.
Many studies showed that the greatest proportion of humans
requiring pretreatment for rabies have been exposed to a rabid
domestic animal.
Human rabies is extremely rare in countries where canine rabies is
controlled by regular vaccination.
According to the WHO, over 30 000 people die each year from
rabies and more than 10 million undergo post- exposure treatment,
having been bitten by a rapid animal.
The disease is a major occupational hazard for veterinarians who
should receive pre-exposure prophylaxis. 921
 Rabies cont’d---
Although individual herds and flocks may suffer many fatalities,
rabies is not of major economic importance in farm animals.
But the disease in human has always been considered fatal.
Pathogenesis
Following the deep introduction of rabies virus by the bite of a rabid
animal, initial virus multiplication occurs in striated muscle cells at
the site.
The following sites provide the virus entry from muscles into the
nervous system.
The neuromuscular spindles
The motor end plates
For example in the olfactory end organ of the nares, neuro -
epithelial cells are the direct contact with the body surface.
These cells extend without interruption into the olfactory bulb of the
brain.
Following entry of the virus into nerve findings, there is invasion of
the brain by passive movement of the virus within axons, first into922
 Rabies cont’d---
the spinal cord then into the brain.
Following entry of rabies virus to the central nervous system
(CNS), usually in the spinal cord, an ascending wave of neuronal
infection and neuronal dysfunction occurs.
The primary lesions produced are in the CNS, spread from the site
of infection occurs only by way of the peripheral nerves.
This method of spread accounts for the extremely variable
incubation period of rabies.
There are many factors that affects the length of incubation period
of rabies like species, dose of the virus etc.
However, the incubation period of rabies varies to a large extent
with the site of the bite.
So that bites on the head usually result in a shorter incubation
period than bites on the extremities.
The severity and the site of the lesions will govern the clinical
picture of rabies which can be
923
 Rabies Cont’d---
Irritative (furious)
Paralytic(dumb)
Gradually ascending paralysis of the hindquarters may be followed
by severe signs of mania, which persist almost until death.
Destruction of spinal neurons results in paralysis.
But when the virus invades the brain, irritation of higher centers
produces
Manias
Excitement
Convulsions
Death is usually due to respiratory paralysis
At death, there are viral inclusions and particles in almost all
neurons in the brain, spinal cord and ganglia.
Electron microscopic examination also shows the presence of the
virus in the cornea
Virus reaches the salivary glands and many other organs in the
same way. 924
 Rabies Cont’d---
But the highly infective nature of saliva arises from passage of the
virus along the olfactory nerve to taste buds other sensory end
organs in the oropharynx, rather than from the salivary glands.
Experimentally the virus reproduced in the adrenal medulla, cornea
and nasal glands.
The virus may be found in milk, in some organs and in fetuses.
But the virus cannot be demonstrated in the blood at any time.
Variations in the major manifestations as mania or paralysis may
depend upon the source of the virus.
Virus from vampire bats almost always causes the paralytic form.
Fixed virus that has been modified by serial intracerebral passage
causes ascending paralysis
In contrast to 'street‘ virus, which more commonly causes the
furious form.
The site of infection and the size of the inoculum may also
influence the clinical course.
925
 Rabies Cont’d---
There is also geographical difference in the proportion of animals
affected by the furious or paralytic form of the disease.
In the Americas most cases are paralytic.
In Africa and India most cases in farm animals are the furious form.
Pathogenesis of rabies can be illustrated as follows

926
 Rabies cont’d---
Pathogenesis
Infection of bite
wounds

Virus multiplies locally in

myocytes for weeks or months

Virus travels up
peripheral nerve to CNS

Virus multiplies in CNS

and causes encephalitis
927
 Rabies cont’d

Virus moves out through the peripheral and

cranial nerves

Then found in salivary glands, skin, mucosal surfaces, gut and most other organs

Saliva is infective for several days/ upto 2weeks/ before clinical signs appears

Death
928
 Rabies cont’d---
Clinical findings
Among farm animals, cattle are the most commonly affected animals
by rabies.
The incubation period of rabies in cattle is about 3 weeks (vary from
2 weeks to several months).
However, the incubation period of rabies may reach 5 and 6 months
in cattle and dogs respectively.
Unvaccinated cattle had shorter incubation and clinical duration of
disease than vaccinated cattle.
Some of the early signs of rabies are non-specific and include
Anorexia
Depression
Mild ataxia
 Rising of body temperature due to muscular activity
Major clinical findings in cattle include
 Excessive salivation (100%)
Behavioral change (100%) 929
 Rabies cont’d---
Muzzle tremors (80%)
Vocalization (bellowing 70%)
 Aggression
 Hyperesthesia and/or hyperexcitability (70 %)
Pharyngeal paralysis (60 %).
The furious form occurred in 70% of cases.
Rabies in cattle may have
1. The paralytic and
2. The furious forms
1. The paralytic( dumb
This form of rabies in cattle is characterized by
Knuckling of the hind fetlocks and sagging
Swaying of the hindquarters while the animal walks
Flaccidity or deviation of the tail to one side
Decreased sensation
930
931
Rabies in dog

932
933
934
935
936
937
 Rabies cont’d---
The above three early signs are best diagnostic criteria for detection
of rabies in cattle.
Drooling of saliva is a constant sign
 Tenesmus with paralysis of the anus resulting in the sucking in and
blowing out of air
 The yawning movements are more accurately described as voiceless
attempts to bellow.
Bulls have paralysis of the penis
Then paralysis follows, animal goes down and unable to rise
Death will be after 48hrs after recumbency or it may have a total
course of 6 - 7 days.
2. The furious (excitative) form
The main clinical signs that you may observe during this form of
rabies are
The animal has tense and alert appearance
Hypersensitive to sounds and movements
Violently attack other animals or inanimate objects 938
 Rabies cont’d---
Violently attack other animals or inanimate objects
Frequently, loud bellowing is usual at this stage.
The sound is characteristically hoarse and the actions are
exaggerated.
Sexual excitement is also common, bulls often attempting to mount
inanimate objects.
There is apparent inability to swallow but others eat normally until
the terminal stages
Termination is characteristically sudden
 Severe signs may be evident for 24 – 48 hours, then the collapses
suddenly, dying within few hours
The course vary from 1 to 6 days
In sheep
The clinical picture is very similar to that seen in cattle.
Rabies occurs usually in a number of animals at one time.
They may attack humans or each other
There is muscle tremor and salivation 939
 Rabies cont’d---
In goats
They become commonly aggressive and continuous bleating( sound
similar to crying) is common.
In horses
 There is unexplained aggressiveness
 Muzzle tremors were the most frequently observed and most
common initial signs.
The most common clinical signs are
Abnormal postures
 Frequent whinnying,
Unexplained aggressiveness
kicking, biting and colic
 Sudden onset of lameness in one limb followed by recumbency the
next day.
 High-stepping gait, ataxia, apparent blindness and violent head-
tossing. 940
 Rabies cont’d---
Lameness in one limb then recumbency the next day
In pigs
The main clinical signs in pigs are
 Excitement and a tendency to attack or dullness and
incoordination.
Affected sows show twitching of the nose and rapid chewing
movements
Excessive salivation
 Clonic convulsions
They may walk backward
Terminally, paralysis and death occurs 12 - 48 hours after the onset
of signs.
Necropsy findings
There is no any specific gross pathology in affected tissues of
rabies.
The main gross pathology that may be observed during rabies are
Emaciated carcass 941
 Rabies cont’d---
The presence of wounds where the animals were bitted.
Hair animals around the head and neck are wetted by saliva
Stomach of affected animals except carnivores animals are mostly
empty.
In carnivores animals you may found other materials other than
food.
In reticulum and omasum of ruminant animals, you may found dry
feed
Brain with its membrane can be edematic with haemorrhagiae.
Histophatologically, you may observe
Encephalomyelitis of brain and spinal chord.
In cytoplasm of neurons you may found inclusion bodies (2–10 µm
in diameter)
These inclusion bodies (2–10 µm in diameter) are called
Negribodies
You may observe Negribodies in neurons of brain, spinal chord,
ganglia, salivary gland and cornea. 942
 Rabies Cont’d---
The histopathological changes of rabies infection include
A non –suppurative encephalomyelitis
Ganglioneuritis
Neuronal necrosis and the formation of glial nodules
Negri bodies are most commonly found in the Purkinje cells of the
cerebellum of ruminants.
Spongiform change has also been reported in the brain of a heifer
infected with rabies virus.
Diagnosis
No antemortem laboratory examination is of diagnostic value.
But tests for lead on blood, urine, and feces may help to eliminate
lead poisoning as a possible diagnosis.
Virus neutralization tests are available
But the presence of antibodies do not have diagnostic importance.
Other available tests are
Passive haemagglutination
Complement fixation 943
 Rabies cont’d---
Radioimmunoassay
Indirect fluorescent antibody staining
These are used to determine immune status rather than as a
diagnostic aids.
Confirmation of rabies depends on careful examination of fresh
brain.
Dogs or other animals, suspected of rabies because of abnormal
behavior should be kept in isolation for 10 days.
The animal is classified as non-rabid if survives and does not show
the typical sign of rabies.
Of course it may still be necessary to carryout a postmortem
examination if the animal die at rabies.
Specimens
The appropriate specimens for virus isolation are
One half of midsagittally sectioned brain
 Cervical spinal cord
Preserve the above specimens in buffer formalin for histopathology,
944
 Rabies Cont’d---
Fluorescent Antibody Test and immuno histochemistry.
NB. Note the zoonotic potential of this organism when handling
carcass and submitting specimens.
The recommended viral laboratory procedures include the following
three tests and it is recommended to use at least two of the tests for
all specimens.
 1. Fluorescent Antibody Test (FAT)
 2. Mice Inoculation
 3. Detection of Negribodies
1. Fluorescent Antibody Test (FAT)
The test is done on impression smears from the brain.
The test is completed approximately within 2 hours.
It is highly accurate and has reliability of over 99%.
2. Mice inoculation
Negative specimen on FAT and still suspicious are inoculated into
experimental mice.
945
Mouse brain is harvested as soon as signs appear and subjected to
 Rabies cont’d---
FAT.
A positive result can be obtained 4 - 7 days after inoculation.
3. Detection of Negribodies
It is histological detection of Negribodies in tissue section.
Result is available in 48 hours.
However, because of the presence false positive diagnoses, the
technique is in some disrepute.
What are Negribodies?
Negribodies are eosionphilic sharply outlined, pathognomonic
inclusion bodies (2–10 µm in diameter).
They are located in the cytoplasm of certain nerve cells containing
the virus of rabies especially in Ammon's horn of the hippocampus,
medulla obligata and cerebellum or gasserian ganglion.
Negribodies are also found in salivary gland, ganglia and cornea.
ELISA is available for the detection of rabies antigen in animals.
An immunoperoxidase test for rabies can be used on formalin-fixed,
paraffin-embedded brain tissues of domestic animals and wild 946
FAT of rabies virus

947
Negribodies under microscope

948
 Rabies cont’d---
animals when fresh tissues are not available.
Immunohistochemical tests may detect the presence of antigen.
Molecular technologies like reverse transcriptase polymerase
Chain Reaction(RT – PCR) test has been found of value in
detecting rabies infection in decomposed brain samples that were
negative by the direct fluorescent antibody test.
Differential diagnosis
The diagnosis of rabies is one of the most difficult and important
duties that a veterinarian is called upon to perform.
The best policy is to handle all suspect animals with extreme care.
If the animal is rabid, it will die and the diagnosis can then be
confirmed by laboratory examination.
Rabies must be differentiated from several diseases of animals
characterized by signs of abnormal mental state or paralysis, or a
combination.
Some of the common diseases are
 1. Acute and subacute lead poisoning in cattle and sheep 949
 Rabies cont’d---
It is easy to differentiate them because there is blindness in lead
poisoning but not in rabies
2. Lactation tetany in cattle and sheep which occurs in lactating
cattle on lush pasture.
3.Vitamin A deficiency occurs in groups of young cattle from 6
months to 18 months of age not receiving adequate carotene intake
or vitamin A supplementation and is characterized by
Polioencephalomalacia in cattle and sheep is characterized by
blindness and opisthotonos
4. Enterotoxemia in sheep is usually confined to lambs on heavy
carbohydrate diets
5. Pregnancy toxemia is a disease of pregnant ewes and is readily
differentiated by the presence of ketonuria.
6. Botulism in cattle
7. In pigs rabies must be differentiated from
 Pseudorabies(Aujesky’s disease)
 From several other diseases of the pigs such as 950
 Rabies cont’d---
Hog( classical) and African swine fever,
 Meningitis associated with Streptococcus suis Haemophilus spp,
Escherichia coli, septicemia and erysipelas
8. In horses rabies must be differentiated from many diseases of the
nervous system including
 Viral encephalomyelitis
Herpes virus myeloencephalopathy
Cerebrospinal nematodiasis
Equine degenerative myeloencephalopathy
Equine protozoal myeloencephalitis
Japanese encephalitis
Botulism

951
 Rabies cont’d---
Treatment
Treatment is not recommended after signs are evident.
In animals, immediately after exposure irrigation of wound with
20% soft soap or zephiran solution prevent establishment of
infection.
Immediate and through washing of all wounds and scratches with
soap and water is the most important measure for preventing rabies
in veterinarians.
Post-exposure vaccination is unlikely to be of value in animals, as
death usually occurs before appreciable immunity has had time to
develop.
Euthanasia of suspect animals must be avoided, particularly if
human exposure has occurred, since the development of the disease
in the animals is necessary to establish a diagnosis.
Anti rabies serum may become available for animal treatment
at some future date.
952
 Rabies cont’d---
Prevention and control measures
The major goal of rabies control in animals is reduction and or
elimination of human rabies.
The most rational approach for this is reducing the prevalence and
incidence in animals.
In developed countries this has been accomplished by vaccination
of dogs and cats.
In farm animals there are three useful techniques for rabies control.
1. Prevention of exposure to virus
2. Pre-exposure vaccination
3. Post – exposure vaccination
1.Prevention of exposure to virus
This can be achieved by controlling access of wildlife species which
are likely to come into contact with the farm livestock.
Foxes accounted for a very large proportion (85% in Europe) of
wildlife rabies.
So that a control program aimed at reducing their population using
953
 Rabies cont’d---
poison or traps was attempted.
This method of population reduction failed to control outbreaks or
to reduce enzootic rabies.
2. Pre-exposure vaccination
The most successful form of rabies prevention is pre-exposure
vaccination.
Farm livestock in endemic areas where clinical cases of rabies occur
commonly should be vaccinated.
For cattle, sheep and horses the primary vaccination is given at 3
months of age and boosters given annually.
In countries where vampire bats are a major vector for rabies in
farm livestock vaccination is necessary.
But in countries those are in developing does not support due to a

cost benefit analysis.

In some countries of the world prevention and control of rabies is
done through vaccination of the wildlife.
Oral immunization on of foxes has resulted in a substantial 954
 Rabies cont’d---
decrease in the number of rabies cases in Europe.
Vampire bats have been vaccinated IM, scarification, oral or aerosol
routes using a vaccine-rabies.
3. Post – exposure vaccination
An effective post -exposure protocol for unvaccinated domestic
animals exposed to rabies includes
 Immediate vaccination against rabies
 Strict isolation period of 90 days a nd administration of booster
vaccinations during the third and eighth weeks of the isolation
period .
The protocol has been effective in dogs, cats, cattle and horses.
Rabies vaccine for domestic animals and wild life
Farm livestock in endemic areas where clinical cases of rabies occur
commonly should be vaccinated.
Almost all rabies vaccines for domestic animals are inactivated.
They are inactivated by binary-ethylenimine and containing
aluminum hydroxide adjuvant.
955
 Rabies cont’d---
The vaccine protects animals for upto 3 years.
Inactivated tissue culture cell vaccines given to cattle result in
neutralizing antibodies in 1 month after the primary vaccination.
A booster given 1 year later increases the titer which are detectable
1 year after the booster.
It is very useful for the control of rabies in cattle where the vampire
bat and wild animals are the main vectors of the disease.
Vaccinal antibodies are present in the colostrum of vaccinated
cows.
It is recommended that where cattle are vaccinated annually, calves
be vaccinated at 4 months of age and again when they are at 10
months of age.
Calves from unvaccinated dams ca n be protected by vaccinating
them at 17 days of age.
Quarantine and biosecurity
The most effective method o f preventing the entry o f rabies into a
country free o f the disease is the imposition o f a quarantine period
956
 Rabies cont’d---
of 4 - 6 months on all imported dogs.
This system has successfully prevented the entry of the disease
into island countries.
But it has obvious limitation in countries that have land boarder.
There fore, vaccination o n two occasions with a n inactivated
vaccine while the animal is still in quarantine for 6 months is the
current recommendation

957
In this portion we give attention to
2.1. Foot and Mouth Disease(FMD)
2.2. Rinder pest (Cattle plague)
2.3. Peste Des Petitis Ruminants (PPR)
2.4. Malignant Catarrhal Fever(MCF)
2.5. Bovine Virus Diarrhea(BVD)

958
 FMD also called Aphthous fever.
Foot and Mouth Disease(FMD) is an extremely contagious viral
disease of all cloven- hoofed animals.
The disease is characterized by
Fever and
Vesicular eruptions in the mouth, on the feet and teats.
Etiology
FMD is caused by virus found under the family of Picornaviridae
and genus Apthovirus
Apthovirus is RNA virus.
It is the smallest among RNA viruses.
FMD virus has seven serotypes, designated as
1. A
2. O
3. C
Southern African Territories (SAT) serotype which includes
4. SAT-1 959
 FMD Cont’d---
5. SAT-2
6. SAT-3
7. Asia-1
There are a number of immunologically and serologically distinct
subtypes with different degrees of virulence, especially within the
A and 0 types.
So far about 80 subtypes have been identified
As there is no cross-immunity between serotypes, immunity to one
type does not confer protection against the others.
This presents difficulties to vaccination programs.
Furthermore, there can be great changes in antigenicity between
developing serotypes; virulence may also change dramatically.
What are the main reasons that make FMD to be difficult to
control?
The presence of many hosts that act as maintenance and amplified
host. For example small ruminants are the maintainace host while
large ruminants are amplified hosts for FMD. 960
 FMD Cont’d---
The presence of many serotypes which do not have cross protection
and are in constant mutation. So that the vaccine must be a potent
vaccine that contains all serotypes those are circulating in a given
area.
The presence of many wild animal which acts as reservoir of the
virus
The ability of the virus to travel more than 250 Km with winds
even across the sea.
Epidemiology
FMD affects all cloven- hoofed domestic and wild animals.
The disease is enzootic in Africa, Asia, South America and parts of
Europe.
The disease can occur in any country.
But Japan, New Zealand and Australia are disease free country.
Western Europe is essentially free of the disease but cattle imported
from Eastern Europe gave rise to an epidemic in many countries of
Western Europe. 961
 FMD Cont’d---
More recently, there was a massive outbreak in Britain in 2001
which spread to Ireland, France and The Netherlands
Spread within the country and to other countries was mostly through
the movement of livestock not showing obvious clinical signs.
There are no reliable figures for the prevalence of FMD in different
countries.
The disease generally occurs in the form of an outbreak that rapidly
spreads from herd to herd before it is controlled.
Of the seven standard types:-
Serotypes O, A and C are prevalent in all continents where the
disease is enzootic.
SAT 1 is found in Africa and Asia
 SAT 2 and SAT 3 are limited to Africa
Asia 1 occurs only' in Asia
Overall, outbreaks of types 0 and A occur more frequently than the
others.
The morbidity rate of FMD in new outbreak areas can reach 962
 FMD Cont’d---
approximately 100%.
However the case fatality rate is usually very low and reaches 2%
in adults and 20% in young animals.
The violent form in exotic animals may cause case fatality rate of up
to 50% in adults.
Source of infection
 Sick animals or animals with latent infection
 Carrier animals
Methods of transmission
FMD virus spreads from one animal to another by
Inhalation
Ingestion
Direct contact
In tropics the most important method to spread FMD virus is by
direct contact between animals which are moving freely for trade or
nomadic cattle.
Air-borne spread is common and the speed and direction of the 963
 FMD Cont’d---
wind are very important factors.
In most favorable circumstances, infection can be wind-borne and
the virus can travel as far as 250 KM and possibly across expanses
of the sea.
All secretions and excretions can be infective before the animal
shows clinical signs of FMD
Among all secretions and excretions vesicular fluid has maximum
FMD virus concentration.
Up to 50% of infected cattle, sheep and goats may become carriers.
But there is no carrier state in pigs.
FMD is spread from one herd to another either
 1.Directly by direct contact between herds.
2. Indirectly by
Inanimate objects
Uncooked and unprocessed meat products
Milk and milk products
Animal products and bi – products other than meat 964
 FMD Cont’d---
Risk factors
The most common risk factors for occurrence of FMD are
1. Host factor
2. Environmental and pathogen factors
1. Host factors
FMD affects all cloven hoofed domestic and wild animals.
The most susceptible animals are cattle and pigs.
But sheep and goats can also be affected.
Elephants, hedgehogs, many rodents and wild ruminants like
antelopes and buffaloes etc. are susceptible to FMD and act as
reservoirs for domestic animals.
2. Environmental and pathogen factors
FMD virus is resistant to common disinfectants and the usual
storage practices of meat and milk in trade.
The virus persists
Over a year in infected premises
For 10 -12 weeks on clothes and feed 965
 FMD Cont’d---
Up to a month on hair
For a month in bull semen stored at -79 0C
The virus is susceptible to
The change of the PH from neutral
Sunlight, to high temperature like boiling
Economic importance of FMD
FMD is probably the world’s most important animal disease in
economic losses.
Losses occur in many ways but the most importants are
Loss of meat and milk production
Reduction of work ability
Interference with movement of livestock and meat between
countries are the most important economic losses
Expense for prevention, control and eradication measures
Pathogenesis
Once infection gains access to the blood, virus is disseminated to
many epidermal sites carried by mononuclear cells. 966
 FMD Cont’d---
But gross lesions develop only in the mouth and feet and to less
extent, on the teats.
Characteristic vesicular lesions develop after an incubation period
of 1 - 21 days (usually 3 - 8 days in most species).
After the end of viremia, animal may recover.
But the virus may persist in pharyngeal area for months or even
years.
Bacterial complications aggravate lesions particularly of the feet and
teats, leading to severe lameness and mastitis.
In young animals the virus frequently causes necrotizing myositis.
Clinical signs
1. In cattle
The incubation period is mostly 3 - 6 days and may reach upto 7
days.
The main clinical signs that can be observed in cattle are
 Fever that can reach upto 410C
Reduction of milk yield 967
 FMD Cont’d---
 Severe dejection( depressed) and anorexia followed by acute
painful stomatitis
Abundant salivation
Smacking of lips and difficulty in chewing
Vesicles and bullas that may have 1-2cm in diameter appear on the
buccal mucosa, dental pad and tongue.
Vesicles rupture after 24 hrs and leaves a raw of painful surface
Concurrently with the oral lesions, vesicles appear on the feet
Rupture of vesicles causes acute discomfort and animal become
grossly lame
The animal is often recumbent with marked painful swelling of the
coronet
Bacterial complication interferes with healing and severe
involvement of deeper structures.
Vesicles on the teats cause severe mastitis
Very rapid loss of condition and fall in milk yield
968
Young calves are more susceptible than adults
969
970
971
Two day old ruptured vesicle (blister) on tongue, lower gum and
low

972
Ruptured oral blister in diseased cow.

973
974
975
976
977
978
979
 FMD Cont’d---
Heavy mortality in calves results due to severe myocardial
damage(Tiger heart).
The occurrence of FMD in sheep, goats and pigs has mild form
But it is important mainly because of transmission of the disease to
cattle.
2. In sheep
The main clinical signs are
Severe sudden lameness
Lies down frequently and is very unwilling to rise
When made to rise stands in a half-crouching position with hind legs
brought well forward, reluctant to move
Blisters may be found on the hoof where the horn joins the skin
which may extend all round the coronet and in the cleft of the foot.
When they burst the horn is separated from the tissues underneath
and hair round the hoof may appear damp
Blisters in the mouth are not always apparent.
 But when they develop they may found on the dental pad and 980
 FMD Cont’d---
sometimes on the tongue.
Diagnosis
Exhaustive laboratory studies are needed to
 Isolate and identification of the agent in tissue or fluid.
FMDV is isolated by inoculation into cell cultures or embryonated
eggs or unweaned mice.
Mostly FMD virus is cultivable on tissue culture and in
embryonated hen eggs,
To identify the isolated FMDV, virus neutralization test with
specific antisera is widely employed.
Then serotyping of isolated virus is done by virus neutralization test
using known antisera.
Serology
The most common serological tests that are used to diagnose FMD
are
1. FAT( direct)
2. ELISA
981
 FMD Cont’d---
2.1. The indirect ELISA can detect the FMDV antigen in
epithelial cells using special monoclonal antibody
The sensitivity of ELISA using special monoclonal antibody is
greater than CFT and approaches virus neutralization test.

982
 FMD Cont’d---
2.2. Serological tests for specific antibody response to FMD
structural or nonstructural proteins by using
Virus neutra lization (VN), a prescribed test for international
trade, the test is serotype specific
 Competitive enzyme linked immunosorbent assay (ELISA)
another prescribed test
 Blocking ELISA
Nonstructural protein (NSF) antibody tests that enable detection of
past or current infection, irrespective of vaccination status.
They are more useful on a herd basis.
The test measures antibody to virus infection-associated antigen
(VIAA) by
Agar gel immunodiffusion (AGIP), or better, antibody to NSPs(
None Structural Proteins)
3. Complement fixation test (CFT)
Direct CFT on epithelial suspension is one of the fastest methods of
making a positive diagnosis, within a few hours. 983
 FMD Cont’d---
But negative samples must be checked in tissue cultures.
Because of the number of false-negatives which occur with the CFT
especially in poorly collected and packaged samples.
4. Nucleic acid recognition methods
These include the reverse transcription polymerase chain reaction
(RT-PCR)
The RT-PCR amplifies fragments of FMD genome in samples and
can be used for typing.
It is more sensitive than ELISA.
A portable real-time RT-PCR that can be performed within 2 h has
been described.
Differential diagnosis
FMD must be differentiated from
Vesicular stomatitis
Swine Vesicular exanthema
Swine vesicular disease
Blue tongue 984
 FMD Cont’d---
Differentiation of acute vesicular diseases
Types of diseases

Species of FMD Vesicular Vesicular Swine Blue tongue

animals stomatitis exanthema vesicular
of swine disease
Cattle + + - - +( occurs rarely)
Pig + + + + -
Sheep and + + - - +
goat
Horse - + - - -

Treatment
Good nursing care and treatment of wounds with mild antiseptic and
antimicrobial ointments are recommended.
Cardiac preparates in the case of heart failure.
A good symptomatic response is reported to the administration of flunixin
meglumine 985
 FMD Cont’d---
Control and preventive measures
In free areas
Rapid identification of outbreaks, quarantine and slaughter of all
affected and exposed animals are effective to prevent the disease.
In endemic areas
Vaccination and quarantine are the bases for control.
The vaccine must be type specific.
Killed trivalent (O, A and C) vaccines are in general use.
Immunity lasts only for 6 - 8 months.
NB. In Ethiopia, there is bivalent vaccine contains O and C strain
which is absorbed into Aluminum Hydroxide gel(Al(OH)3
concentrated and inactivated with 0.3% of formaldehyde.
 It is presented as 50 ml and 100 ml vials of suspension.
 Route of administration: - S/C in dewlap in cattle and in the inner
thigh of animals in small ruminant animals.
 Dose : Cattle - 4ml/ animal above 6 months of age
Small ruminants 2ml/ animal above 6 months of age 986
 FMD Cont’d---
 Revaccinate again after 6 months and then after every 6 – 8
months.

Rinderpest is an acute, highly contagious viral disease of ruminants

and swine.
It is characterized by
High fever
Necrotic stomatitis
Bloody diarrhea with high mortality

The rinderpest virus belongs to

Family of Paramyxoviridae
The genus Morbillivirus
It is RNA virus
RP virus is quite fragile
RP virus is antigenically related to
Peste des Petits Ruminants (PPR) virus in sheep and goats 987
 Rinder pest Cont’d---
Canine distemper virus in dogs
Measles virus in humans
Phocine distemper virus in seals
The virus occur as many strains with considerable variation in
virulence.
But all strains are immunologically identical.
That means, vaccination for one strain confer protection against all
other strains.
Epidemiology
Rinderpest (cattle plague) was a most devastating disease spread
from Asia to Europe, the Middle East and Africa, usually as a sequel
to wars.
The need to combat the disease was instrumental in the
establishment of the first veterinary school in 1762 in Lyon, France.
Most countries today claim to be free of rinderpest.
At the end of the last century, the disease was still endemic in parts
of Asia and in a small area bordering on Kenya and Somalia. 988
 Rinder pest Cont’d---
The most recent outbreaks occurred in Kenya and (1993 - 97,
2003) and in Pakistan.
All ruminants and pigs are susceptible to infection with rinderpest
virus.
Natural infection occurs commonly only in domestic cattle, buffalo
and yak.
Cattle and pigs are susceptible to infection with rinderpest virus.
But in some outbreaks, it may affect sheep and goats.
One humped camels become infected, but show no clinical signs.
Wildlife are often affected during outbreaks and the infection usually
spreads to them from infected cattle.
When epidemics occur in highly susceptible populations, the
morbidity and case-fatality rates approximate 100% and 50% (25-
90%) respectively.
Method of transmission
Close contact between infected and non-infected animals is
necessary for the spread. 989
 Rinder pest cont’d---
Because the virus does not survive long outside the host.
All secretions and feces of infected animals are contagious
throughout the course of the disease.
Transmission occurs through
Contaminated feed and/or
By inhalation of aerosol.
European breeds of pigs, sheep, goats, camels and wild ruminants
may act as source of virus for cattle.
Risk factors
The risk factors for occurrence of rinder pests are
1. Host factors
2. Environmental and pathogen factors
1. Host factor
Cattle and buffalo of all ages are susceptible.
Amongst races of cattle, European breeds are more susceptible than
zebu cattle.
990
2. Environmental and pathogen factors
 Rinder pest Cont’d---
The highest virus concentration is seen at the highest temperature
reaction of the affected animals.
So the risk of transmission of the disease is greatest during the
febrile stage.
The virus is very susceptible to external influence and does not
persist outside the host body for more than a few hours.
It is readily destroyed by heat, drying and most disinfectants.
Pathogenesis
The virus is inhaled in infected droplets, penetrates epithelium of the
upper respiratory tract and multiplies in the tonsils and regional
lymph nodes.
Then it enters the blood in mononuclear cells and disseminated to
other lymphoid organs, the lungs and epithelial cells of mucous
membranes.
RP virus has high degree of affinity for lymphoid tissue and alimentary mucosa.

There is striking destruction of lymphocytes in tissues causing

leucopenia. 991
 Rinder pest Cont’d---
The focal, necrotic stomatitis and enteritis, which are characteristic
of the disease, are the direct result of viral infection and replication
in epithelial cells in the alimentary tract.
Death is usually from severe dehydration resulting from diarrhea.
In less acute cases, death may be from activated latent parasitic or
bacterial infections which are exacerbated by immunosuppression as
a result of destruction of lymphoid organs by the virus.
Clinical findings
The incubation period of RP is 3 to 15 days.
The main clinical signs during rinder pest are
 Sudden onset of fever, depression and anorexia
The muzzle is dry and mucous membranes are congested
 Within days there will be oral erosions where necrotic foci have
sloughed
Nasal and ocular discharge
Purulent lacrimation is a constant sign
992
993
994
995
996
997
998
999
1000
1001
1002
1003
1004
1005
1006
1007
1008
 Rinder pest Cont’d---
Diarrhea is severe and blood may be present
Dehydration and emaciation lead to death.
Necropsy findings
Lesions are found mainly in the alimentary tract and lymphoid
tissue.
The main necropsy findings in rinder pest are
Oral erosions with red floor
Edema and congestion of abomasum
Ulcers and hemorrhagic to necrotic in Peyer’s patches of small
intestine
Lymph nodes have necrotic germinal centers
Carcass emaciated, dehydrated and soiled with fetid feces
Severe changes also occur in the mucosa and Peyer’s patches of the
large intestine which is hemorrhagic necrosis
 Zones of hemorrhage and erythema running transversely across the
colonic mucosa produces a characteristic striped appearance, the so
called ‘zebra stripes’. 1009
Hyperemia of small intestine

1010
Hyperemia of the large intestine

1011
Haemorrhagic mesenteric lymph nodes

1012
1013
 Rinder pest Cont’d---
The disease in sheep and goats is mild or sub- clinical.
The disease in sheep and goats is mild or sub- clinical.
Diagnosis
As usual diagnose of rinder pest is done based on
 Epidemiology of the disease
 Clinical signs
Postmortem findings
The above three tests give the tentative diagnose of the disease.
However the confirmatory test is done by virus isolation and
identification in viral laboratories.
Laboratory diagnosis
Laboratory confirmation of RP can be accomplished by
1. Antigen detection and virus isolation
For antigen detection and virus isolation, the key to diagnostic
success is 1014
 Rinder pest Cont’d---
The collection of suitable samples at the optimum time (3 - 5
days)after fever commences) from many animals rather than many
samples from one sick or dead animal.
Use as antigen the needle biopsy of lymph nodes and detect by
AGID with specific antisera
Other satisfactory methods of detecting rinderpest antigen in feces,
buccal scrapings and ocular and nasal discharges in the early stages
of the disease are
 Complement fixation test
Immunofluorescence
Immunohistochemistry
Passive haemagglutination tests
The RPV can be detected by using Reverse transcriptase PCR and
DNA probes.
Virus isolation
Virus can be isolated using tissue culture or using continuous
growing cell lines of bovine T-lymphocytes. 1015
 Rinder pest Cont’d---
Virus identification can be done by using virus neutralization test.
Antibody detection is done by
CFT
ELISA
In direct FAT
Immunoperoxidase
Differential diagnosis
Rinder pest must be differentiated from various viral, bacteria and
other diseases of cattle and small ruminants.
Diseases that appear similar clinically with RP are
FMD
Malignant catarrhal fever (MCF)
Bovine virus diarrhea (BVD)
Infectious bovine rhinotrachitis(IRT)
PPR
MCF rarely affects many animals in one herd and is characterized
by specific eye lesions and nervous signs. 1016
 Rinder pest Cont’d---
BVD occurs either in explosive outbreaks like rinderpest.
But the mortality rate is low or it occurs sporadically.
Infectious bovine rhinotrachitis may produce similar mucosal
lesions.
But there is no diarrhea and the mortality rate is too low in IRT.
In sheep and goats, PPR presents the greatest problem in
differentiation
But pneumonia is the usually a feature of PPR but not in rinderpest.
If only sheep and goats a re affected in an outbreak, it is most likely
PPR.
Treatment
Treatment is ineffective and should not be undertaken because of the
danger of disseminating the disease.
Prevention and control
Since there is only one known serotype and protection after infection
or vaccination is life-long.
Vaccination of cattle with a live cell culture attenuated virus is1017
an
 Rinder pest Cont’d---
effective means of control of rinderpest.
NB. In Ethiopia there is a vaccine for rinder pest.
It is freeze – dried lyophilized virus vaccine of Kebete – strain.
The vaccine contains 2.5 TCID 50 and presented in vials of 20 ml of 100
doses.
Reconstitute the vaccine with 100 ml of sterile saline water.
Dose 1ml/ animal above 6 months of age.
Immunity develops 8 days after vaccination and lasts and may lasts for
life.
Control of epidemics in areas where the disease is not endemic involves
Quarantine
Slaughter of
 Infected animals
Contact animals
Other contact ruminants
Disinfection of premises is essential in controlling the disease.

1018
 Pest des petitis ruminants(PR R) is an acute highly contagious viral
disease of goats and sheep characterized by
 Fever
Anorexia
Necrotic stomatitis
Diarrhea
Oculo - nasal purulent discharge and respiratory distress
Etiology
PPR is caused by RNA virus called Morbillivirus in the family of
Paramyxoviridae.
As rinder pest virus, PPR virus is closely related to
Rinderpest virus
Canine distemper virus
 Phocine distemper virus
Measles virus
1019
 PPR cont’d---
Four lineages of PPRV have been identified
These lineages are
Lineage 1 and 2 viruses in west Africa
Lineage 3 in east Africa, Arabia and southern India and
Lineage 4 in the Middle East and Asia subcontinent, reaching east
as far as Nepal and Bangladesh.
All four lineages have been shown to be genetically distinct from
the rinderpest virus.
There is some doubt that PPRV might have evolved from goat-
adapted rinderpest vaccines.
Epidemiology
It is contagious disease of goats and sheep endemic in west and
central Africa
There is a continuous outbreaks of PRR in northeastern Africa,
Middle East and Asia.
The disease occurs in goats and less often in sheep.
Infection rates in enzootic areas are generally high (above 50%) 1020
and
 PPR cont’d---
can be upto 90% of the flock during outbreaks.
The disease, however, is more severe in goats than in sheep and is
rapidly fatal in young animals.
Case fatality rates are also much higher in goats (55 - 85%) than in
sheep (less than 10%).
There is no significant seasonal variation in the prevalence of the
disease.
But since maternal antibodies are lost at about 4 months of age, the
number of susceptible animals is likely to increase 3 - 4 months after
peak kidding and lambing seasons.
PPR is an important economical disease.
As it affects goats and sheep mostly in West Africa and possibly in
all countries where the disease occurs.
In many of those countries, these animals are a major source of
animal protein.
The virus of PPR does not affect humans, so it does not have
zoonotic implication. 1021
 PPR cont’d---
Methods of transmission
Close contact with infected animals or contaminated fomites is
required for disease to spread.
 Large amounts of virus are present in all body secretions and
excretions, especially in diarrheic feces.
Infection occurs mainly by inhalation.
 However infection could also occur through the conjunctiva and
oral mucosa.
Risk factors
The risk factors for PPR are
Species
Age of animals
Breed
Goats are more susceptible than sheep.
Kids over 4 months to under 1 year of age are most susceptible.
Sahelian breeds of sheep and goats are believed to be more resistant
than the dwarf breeds in the humid and sub-humid zones of West1022
 PPR cont’d---
Africa.
Risk of outbreak greatly increased when:
A new stock is introduced or
Animals are returned unsold from markets
NB. Recovered animals have lifelong lasting immunity.
Pathogenesis
Virus penetrates the retropharyngeal mucosa setsup viremia and
specifically damages
The respiratory system
The alimentary system
Lymphoid system
As the result infected cells undergo necrosis .
In respiratory mucosa and lungs there is also proliferation of cells.
Virulent strains cause death from severe diarrhea and dehydration,
particularly in young goats before respiratory lesions become severe
or is hastened by concurrent diseases such as pneumonic
pasteurellosis, coccidiosis or coliform enteritis. 1023
Ly
 PPR cont’d---
Mphoid

In others death is hastened by concurrent diseases such as

pneumonic pasteurellosis and coccidiosis.
Lymphoid necrosis is not as marked in this disease as in rinderpest.
Clinical findings
The disease can occur
Acute or
Subacute.
The acute form is seen mainly in goats and is similar to rinderpest in
cattle.
But severe respiratory distress is a common feature of PPR and signs
appear 3 - 6 days after contact with infected animal.
The main clinical features of PPR in acute form of PPR are
High fever (above 40oC) accompanied by dullness
 Sneezing and serous discharge from eye and nose
 A day or two days later, discrete necrotic lesions develop in mouth
and extend over the entire oral mucosa forming diphtheritic
1024
plagues.
PPR in a goat: inflamed (reddened) eye membranes

1025
PPR in a goat: purulent eye and nose discharges

1026
 PPR cont’d---
PPR in a goat: early mouth lesions showing areas of dead cells
Early pale, grey areas of dead cells on the gums.

1027
PPR in a goat: later mouth lesions
The membrane lining the mouth is completely obscured by a thick
cheesy material; shallow erosions are found underneath the dead
surface cells.

1028
PPR in a goat: swollen, eroded lips
The lips are swollen, oedematous and show areas of
erosion.

1029
PPR in a goat: signs of diarrhoea
The hindquarters are soiled with liquid faeces.

1030
 PPR cont’d---
There is profound halitosis and the animal is unable to eat because
of a sore mouth and swollen lips.
Nasal and ocular discharges become mucopurulent
 Exudates dry up, matting the eyelids and partially occluding the
external nares
Diarrhea develops after 3 - 4 days of onset of fever
Diarrhea is profuse and may be mucoid or blood tinged
Dyspnea and coughing occur later
Respiratory signs are aggravated by secondary bacterial pneumonia.
Death occurs within a week of onset of illness
Erosions have been described in the vulva and prepuce.
Abortions have been reported during outbreaks.
 Sub acute form of PPR is more common in sheep but also occur
rarely in goats.
Signs and lesions are less marked in subacute form of PPR
A few animals may die with in 2weeks, however most recovers
Contagious ecthyma (Orf) may complicate the labial lesions or 1031
 PPR cont’d---
develop in surviving animals.
Necropsy findings
The necropsy findings during PPR are
The carcass is severely dehydrated
The hindquarters are soiled with fluid feces
There is crusts of exudate are present around eyes, nose and lips.
Discrete or extensive areas of erosion, necrosis, and ulceration are
present in the oral mucosa, pharynx, and upper esophagus and may
extend to the abomasum and distal small intestine.
 Hemorrhagic ulceration is marked in the ileocaecal region, colon
and rectum where they produce typical 'zebra stripes'.
Regional lymph nodes are enlarged and wet.
 Spleen may be enlarged.
Severe lesions are often present throughout the respiratory tract that
include 1032
PR in a sheep: advanced pneumonia

1033
PR in a goat: "zebra striping" in the large intestine

1034
 PPR cont’d---
A mucopurulent exudate extends from the nasal opening to the
larynx.
Trachea and bronchi may be hyperemic and contain froth due to
pulmonary congestion and edema.
Diagnosis
Diagnose of PPR can be based on the epidemiology, clinical,
necropsy findings and laboratory findings.
Other diseases that cause diarrhea or pneumonia pose a diagnostic
challenge.
Clinical and postmortem findings of stomatitis, enteritis and
pneumonia are typical for PPR.
As diarrhea develops, there is a progressive haemoconcentration(
increased PCV value) and low serum sodium and potassium.
For diagnostic purpose, specimens should be collected from several
live animals and should include
Swabs of conjunctival, nasal and buccal mucosae
1035
 PPR cont’d---
Whole blood in anticoagulant for virus isolation
At necropsy, the following specimens should be collected for
virology and histopathology.
Lungs
Small and large intestines
Oral mucosa
Tonsil
Mesenteric lymph nodes.
The laboratory tests are
1. Virus isolation
2. Virus identification by using
Molecular techniques
Serology
PPR virus is isolated by using Vero cells.
Isolated virus can be identified by
Reverse transcription-polymerase chain reaction (RT – PCR)
1036
 PPR cont’d---
Serological tests with monoclonal antibodies like
Virus neutralization test (VNT)
Agar – gel immuno diffusion(AGID)
Complement fixation test(CFT)
Enzyme – linked immunosorbent assay specially Competitive
ELISA( C- ELISA) specific for the nucleocapsid (N) or
haemagglutinin (H) proteins of PPR
The presence of past infection of PPR can be known by serological
test that are mentioned above.
NB. The reverse transcription-polymerase chain reaction (RT -
PCR) has been reported to be more rapid and far more sensitive
than conventional identification of the PPR virus on Vero cells by
using VNT.

1037
Differential diagnosis
Other diseases that cause diarrhea or pneumonia in sheep and goats
may pose a diagnostic challenge
But a history of recent introduction of new stock and the clinical and
postmortem findings of stomatitis, enteritis and syncytial giant cell
pneumonia are typical for PPR.
Laboratory tests are required to rule out rinderpest.
Other diseases to be considered are
Heart water
Pneumonic pasteurellosis
Contagious Caprine Plueropneumonia in goats
Contagious Bovine Plueropneumonia
 Helminthiasis
Coccidiosis
Contagious ecthyma
Possibly Nairobi sheep disease
Treatment
1038
Valuable for sick animals in the early stages of the disease.
 PPR cont’d---
Sick animals must be isolated from herds as soon as possible.
Then give hyperimmune serum which may be obtained from cattle
hyper immunized against rinderpest.
Supportive treatment includes fluid therapy for dehydration and
antibiotics to prevent secondary bacterial infections.
Lesions around the eyes, nostrils and mouth should be cleaned and
good nursing provided.
Preventive and control measures
The disease is effectively prevented by not introducing new stock
from unknown sources.
Animals returned from markets should be segregated from other
herds.
Vaccine is a available to prevent and control PPR.
In Ethiopia PPR vaccine is available and called lyophilized freeze –
dried PPR virus vaccine with the minimum titer of 2.5 TCID 50
with 20 ml of vial of 100 dose.
1039
 PPR cont’d---
Reconstitute with saline.
Dose and route of administration :- For goats and sheep 1 ml SC in
the inner thigh of animals.
Vaccinate all sheep and goat above 6 months of age.
The immunity lasts one year so that annual vaccination of sheep and
goat is required.

1040
 It is infectious and non – contagious viral disease of mainly cattle
and buffalo characterized by
 Fever
Inflammation of the respiratory and GIT mucosae
 Infection of eye and CNS
Etiology
MCF is caused by DNA virus in the family of Herpes viridae and
genus Herpes virus.
The virus replicates in cell culture of thyroid gland, adrenaline
gland and lung and cause CPE.
Epidemiology
In natural condition MCF affects mainly cattle and buffalo.
However, sheep, goat, pigs, deer, giraffe and antelopes are
susceptible to the virus.
All animals except cattle and buffalos are carrier animals and aret
he main source of infection for cattle and buffalo. 1041
 MCF Cont’d---
The epidemiology of MCF is not well studied and many scientists
consider animals except cattle and buffalos are carrier animals and
are the main source of infection for cattle and buffalo.
So that source of infection in MCF infection are
Sick animals
Carrier animals(latent form of infection)
Old animals are more susceptible than young animals.
Bulls and oxen are more susceptible than cows.
The disease mostly occurs sporadically and very rarely it occur as
epidemics(epizootics).
Transmission
Wild animals and sheep are carriers for MCFV.
Infection of new born wild animals can be occurred horizontally
and occasional intrauterine transmission may occur.
So that infected young wild animals upto the age of about 4 months
have viremia and shed virus in ocular and nasal secretions.
1042
 MCF Cont’d---
The disease is transmitted from wild animals to cattle by
Inhalation of aerosol or
Ingestion of pasture contaminated by virus excreted by young wild
animal in nasal and ocular discharges.
However, there is occurrence MCF outbreak in feed lots where the
cattle do not have contact even with sheep.
This leads us to suspect that the MCFV persistent on inanimate
fomites and transmitted to the farm.
But the virus is a most fragile one and this seems unlikely.
The observation that some recovered cattle show
A persistent viremia for many months suggests that carrier cattle
may be the source of these carryover infections.
In addition, the virus, detected by PCR, has been demonstrated in
cattle and farmed deer with no evidence of MCF disease.
It is possible that stress could activate a latent infection in animals
with no sheep contact.
Stress factors that aggravate the latent infection of cattle to disease
1043
 MCF Cont’d---
in feed lots are
Law temperature
High humidity
Poor ventilation
Un balanced ration
Housing of cattle and sheep together
Risk factors
The risk factors for the occurrence of MCF are
1. Environmental risk factor
2. Animal risk factors
1. Environmental risk factor
The disease shows the greatest incidence in late summer and
spring months.
There have been suggestions that
Copper deficiency and/or
 Exposure to bracken fern might be environmental stressors that
predispose the expression of the disease in cattle. 1044
Bracken Fern Poisoning of Cattle

1045
 MCF Cont’d---
2. Animal risk factors
Amongst domestic animals, all ages, races and breeds of cattle are
equally susceptible.
But bison and deer are more susceptible and suffer a more severe
form of the disease than do cattle.
Pathogenesis
It is believed that the pathogenesis of this disease is the result of
direct virus-cell interactions or
Perhaps immune-mediated responses directed against infected cells
CD8+ T- lymphocytes are the predominant cells associated with the
vascular lesions.
MCF is a fatal, multisystemic disease characterized by
Lymphoid proliferation and infiltration
 Widespread vascular epithelial and mesothelial lesions, which are
morphologically associated with lymphoid cells.
Clinical findings
The incubation period in natural infection varies from 3 - 8 weeks.
1046
 MCF Cont’d---
MCF is described as occurring in a number of forms
1. Peracute form
2. Alimentary tract form
3. Common 'head and eye' form
4. Mild form
1. Peracute and alimentary tract forms
In the peracute form the disease runs a short course of 1 - 3 days.
There is usually
 High fever
Dyspnea
An acute gastroenteritis
The alimentally tract form resembles the 'head and eye' form, except
that there is marked diarrhea and only minor eye changes consisting
of conjunctivitis.
2. Common 'head and eye' form
There is a sudden onset of the following symptoms
Extreme dejection 1047
MCF in cattle

1048
1049
MCF cont’d---

1050
1051
 MCF Cont’d---
In addition to the above clinical findings there is
Superficial necrosis is evident in the anterior nasal mucosa and
on the buccal mucosa.
As the result there is
Discrete local areas of necrosis appear on the hard palate, gums
 The mouth is painful at this time and the animal moves its jaws
carefully
The mouth is painfully and with a smacking sound.
The mucosa of oral cavity as a whole is fragile and splits easily.
 The mouth and tongue are slippery and the mouth is hard to
open.
The erosive mucosal lesions may be localized or diffuse and
they may occur on the:-
Hard palate
Dorsum of the tongue
1052
1053
 MCF Cont’d---
Gums below the incisors
Commissars of the mouth
Inside the lips.
The cheek papillae inside the mouth are hemorrhagic, especially at
the tips
At this stage there is excessive salivation which is ropy and bubbly,
hanging from the lips.
3. Mild form
The mild form of MCF occurs very rarely in cattle.
In this form of MCF, there is
 Transient fever and
 Mild erosions on the oral and nasal mucosae.
Mild disease may be followed by complete recovery, recovery
with recrudescence or chronic MCF.
Very rarely there is a typical form of MCF called MCF with nervous
signs.
It is characterized by 1054
 MCF Cont’d---
Weakness particularly in one leg
Incoordination
Demented appearance
 Muscle tremor
 Nystagmus
Paralysis and convulsions may occur in the final stages
Necropsy findings
Lesions with minor degrees of haemorrhage and erythema, through
extensive, severe inflammation to discrete ulcers are found in the
mouth, nasal cavities and pharynx.
These lesions may be shallow and almost imperceptible or deeper
and covered by cheesy diphtheritic deposits.
There is erosion of the tips of the cheek papillae especially at the
commissures
Longitudinal, shallow erosions are present in the esophagus.
Similar lesions are present in the mucosa of forestomachs and
abomasum. 1055
1056
 MCF Cont’d---
Catarrhal enteritis of moderate degree and swelling and ulceration
of the Peyer's patches are constant.
Similar lesions to those in the oral cavity are found sometimes in
the bronchi.
But the lungs are not usually involved except for occasional
emphysema or secondary pneumonia.
The liver is swollen
Severe hemorrhage may be visible in the urinary bladder.
All lymph nodes are swollen, edematous and often hemorrhagic.
Diagnosis
MCF is diagnosed with complex method comprises by considering
The epidemiology of the disease
 Clinical findings
Necropsy findings
These give the tentative diagnose of the disease.
Confirmatory diagnose is done by laboratory diagnostic method.
For laboratory diagnose the specimens of choice are different. 1057
 MCF Cont’d---
For virology and molecular techniques
Lymph node
Spleen
Liver
Peyer's patch of intestine
For histopathology: - Brain, lymph node, alimentary tract
mucosa(pharynx, esophagus, rumen and Peyer's patch) liver, adrenal
gland, kidney and urinary bladder that are fixed in buffer formalin.
Virus isolation is not practical in MCF diagnosis.
Because of the instability of the virus and the fact that some strains
do not replicate in cell culture.
There are a number of serological tests that can be used.
But they have limited value for diagnosis of clinical cases.
Because only a small percentage of animals seroconvert and do so
late in the course of the disease.
The antibody titer is low and there is cross reaction with other
herpes viruses. 1058
 MCF Cont’d---
Among serological tests competitive- ELISA using a monoclonal
antibody to a broadly conserved epitope of the MCF virus is widely
used and has largely replaced other serological tests.
The most widely used to diagnose MCF is detection of viral nucleic
acid by PCR.
Detection of viral nucleic acid by PCR techniques is the current
accepted diagnostic technique for MCF.
Differential diagnosis
MCF must be differentiated from
Oral mucosal disease is called stomatitis of non – infectious nature
Infectious bovine rhinotrachitis(IRT)
Sporadic bovine encephalomyelitis
Rinderpest
Treatment
Treatment of affected animals is unlikely to influence the course of
the disease.
But it is possible to use 1059
 MCF Cont’d---
Broad spectrum antibiotics and appropriate antiseptics to dress the
erosions in oral cavity.
Non-steroidal anti-inflammatory may ease the discomfort.
Prevention and control measures
Because of the field observation that sheep are important in the
spread of MCF.
So that separation of cattle and sheep herds is recommended.
The introduction of sheep from areas where the disease has
occurred to farms with cattle should be avoided.
Attempts to immunize cattle with live or inactivated culture
vaccines with Freund's incomplete adjuvant
But they do not provide protection against experimental challenge
or natural challenge by exposure.
High and persistent levels of virus-neutralizing antibody are
demonstrable following vaccination.
But humoral mechanisms are probably not important in
1060
 MCF Cont’d---
determining resistance to infection with the virulent virus.
So that there is no any effective vaccine to prevent and to control
MCF.

1061
 Bovine viral diarrhoea(BVD)has also another name mucosal
disease or bovine pestivirus disease complex.
It is viral contagious disease of cattle characterized by
Fever
Erosal– ulceratative inflammation of the mucous membrane of
digestive and respiratory systems which is accompanied by
diarrhoea, rhinitis, conjunctivitis and rarely with abortion.
Etiology
The positive agents of BVD are classified in the virus family of
Flaviviridae and genus Pestivirus.
The bovine viral diarrhea virus (BVDV) is one of three pestiviruses:
 1. Bovine viral diarrhea virus (also known as mucosal disease virus
or bovine pestivirus)
 2. Border disease virus of sheep
 3. Hog cholera virus (also called European or classical swine fever
virus).
1062
 BVD Cont’d---
Cross - infection between species can be achieved experimentally
and has been demonstrated in field infections.
Among the ruminant pestiviruses there are two biotypes in the
BVDV, depending on their effect on tissue culture cells designated
as
 Non-cytopathic (NCP)
 Cytopathic (CP)
The non – cytopathic type is the most common and most important.
Because only the non-cytopathic type crosses the placenta, invades
the fetus and establishes persistent infection in the fetus, which is
crucial for spread of the virus.
In contrast, the cytopathic biotype of the virus is usually associated
with only mucosal disease in animals already persistently infected
with the non-cytopathic biotype.
Both biotypes can be isolated from animals dying of mucosal
disease.
There is evidence that the cytopathic biotype evolves by mutation1063
 BVD Cont’d---
from the non cytopathic biotype within PI( persistent infected)
animals.
Only non cytopathic BVDVs cause severe acute bovine virus
diarrhea.
Epidemiology
Diseases associated with BVDV have been recorded in most
countries where cattle are raised.
Since the virus is immunosuppressive, it is now clear mucosal
disease occurs only in PI( Persistent infected) animals .
As the result of a congenital infection with a non-cytopathic
strain of the virus acquired in early fetal life.
These animals remain specifically immunotolerant to the
homologous strain of the BVDV throughout postnatal life.
Peracute enteric form of the disease and thrombocytopenia in
young and adult immunocompetent animals infected with highly
virulent strains of the virus were recognized.
1064
 BVD Cont’d---
This was the first indication that mucosal disease could occur in
immunocompetent animals as a result of postnatal infection.
Young cattle which are persistently infected with a non-cytopathic
strain of the virus are the major source of infection in a herd.
Mucosal disease in PI(persistent infected) immunotolerant
seronegative animals occurs in all classes of cattle mostly
Between 6 and 24 months of age
 Rarely in calves as young as 4 months of age
Cattle older than 2 years of age
Outbreaks of the recently recognized peracute BVD occurs in
immunocompetent non-PI(persistent infected) animals are
characterized by
High case rate among all clinically affected animals
Morbidity rates may reach 40%
 Mortality rates 20 %

1065
 BVD Cont’d---
Clinically sick animals called PI(persistent infected viremic animal)
Animals with latent infection
Methods of transmission
The virus can be excreted from sick and latent infected animals
together with nasal discharge, saliva, semen, feces, urine, tears and
milk
Each of them would allow the wide dissemination of the virus.
The virus can be transmitted to susceptible animals by
 Direct contact
 Indirect contact
Some the direct method of transmissions are
Direct contact between animals
Transplacental(transmission to the fetus)
Nose-to-nose contact is an effective method of transmitting the virus
from PI(persistent infected) to susceptible animals.
Thus PI( persistent infected) animals may introduce the infection
into a herd when:- 1066
 BVD Cont’d---
 Infected animals are mixed with susceptible animals at the time of
breeding or pasture or marketing or transportation
 Under conditions requiring emergency movement because of
drought or flood etc
 Accidental mixing of a PI bull with susceptible breeding females
during the breeding season.
 Introduction of an unknown persistently infected cow
The fetus can be infected by transplacental transmission of the
virus from the infected dam, whether the dam is transiently or
persistently infected.
Epidemics of abortion and congenital defects of calves due to
BVDV have occurred when
Transplacental virus infection of the fetuses of cows in the first
trimester
 Previously virus-free herds has followed the introduction of
BVDV-infected animals.
PI bulls may also introduce the virus into artificial breeding units.
1067
 BVD Cont’d---
A PI bull can shed the virus in his semen for a long period and if
introduced into a susceptible herd could have immediate
undesirable effects on reproductive performance.
However, PI bulls may have acceptable semen quality and fertility.
Previously unexposed heifers have been shown not to conceive to
service by a PI bull until they have seroconverted.
The virus may be transmitted in cattle by artificial insemination
with semen from a PI bull.
The use of semen from a transiently infected bull has the potential
to introduce pestivirus infection into a group or herd of susceptible
animals.
But the conception rates are usually within normal ranges.
Some of the indirect means of BVDV transmissions are
 Airborne transmission
 By blood feeding flies
By different fomites like per rectum using the same glove;
hypodermic treatment and vaccination needle 1068
 BVD Cont’d---
Risk factors
Some of the risk factors in the occurrence of BVD are
Animal risk factors
Environmental and managemental risk factors
Pathogen risk factors
In general, young cattle are most susceptible to BVDV infection.
But adult cattle may develop severe disease if infected with the
highly virulent genotypes of the virus.
Unvaccinated animals, improperly vaccinated animals, or animals
whose immune status has waned are most susceptible to infection .
Vaccinated animals may be susceptible if field isolates of the virus
are distinct from those used in the vaccine.
PI animals are susceptible to the development of mucosal disease
following superinfection with the cytopathic virus.
They are also susceptible to other infectious diseases such as
pneumonia.
1069
 BVD Cont’d---
Environmental and management risk factors
The major management risk factors are
The introduction of PI animals into a susceptible herd
 The failure of a vaccination program or an inadequate vaccination
program.
Pathogen risk factors
Several isolates have been identified that are antigenically related.
But there may be antigenic variants that are immunologically
distinct.
In addition to antigenic diversity among strains, there are major
molecular differences between the same strains.
Monoclonal antibodies that have neutralizing activity differentiate
BVD viruses into several groups.
The antigenic variability of this virus may also explain the wide
range of lesions and disease complexes which have been observed.
The antigenic variability of this virus may also explain the wide
range of lesions and disease complexes which have been observed. 1070
 BVD Cont’d---
Economic importance
BVD disease complex is considered as one of infectious diseases
known as production limiting diseases in dairy herds.
The economic losses associated with the introduction of the BVDV
into a susceptible herd of pregnant cattle are due to
Abortion
Congenital defects
Stillbirths
Increased neonatal mortality
 Increased occurrence of other infectious diseases
Prenatal and postnatal growth retardation
Suboptimal reproductive performance due to infertility, deaths from
mucosal disease and the early disposal of PI animals.
Large losses due to fetal infection occur during the first 2 - 3 years
following introduction of infection to a susceptible herd.
Pathogenesis
Pathogenesis depends on multiple interactive factors. 1071
 BVD Cont’d---
Host factors which influence the clinical outcome of BVDV which
includes
Whether the host is immunocompetent or immunotolerant to the
virus
Age of the animal
Transplacental infection and gestational age of the fetus
Induction of immune tolerance in the fetus and the emergence of
fetal immune competence at about 180 days of gestation
Immune status (passively derived or actively derived from previous
infection or vaccination)
Presence of stressors
Routes of infection for BVDVs are the mucous membrane of GIT
and breathing systems.
The virus reaches into tonsil of the organism where it reproduces.
After 1 – 4 days develops viremia.
The virus infected all organs and affects them.
1072
 BVD Cont’d---
During viremia develops fever.
BVDV is an immunosuppressive virus.
Because it destructs
Leukocytes and cause leukopenia
 Reduces the phagocytic activity of neutrophiles and
macrophages
Such deep immunological suppression increases susceptibility
of the animals to other infection.
Immunological suppression occurs in both clinical and
persistent infection of BVD.
BVDVs affects the epithelium of the mucous membrane and
cause degenerative change and cause necrosis and erosion.
Inflammation the mucous membrane of GIT results diarrhea,
dehydration and electrolyte – acid base in balance.
The virus passes through placenta to fetus causes death fetuses
and abortion.
1073
 BVD Cont’d---
Clinical findings
Incubation period of BVD is 2 – 14 days.
Forms and severity of the disease depends on
Virulence and dose of the virus
Immunological status of the organism specially the presence or
absence of specific anti - BVDV antibody.
Clinically BVD has the following forms
Acute
Subacute
Chronic
Latent
In acute form of BVD the main clinical forms are
Fever(40.5 – 42.40C)
Depression
Anorexia
Hyperemia the mucous membrane of nasal cavity with mucosal
nasal discharge 1074
 BVD Cont’d---
On mucous membrane of oral cavity you may observe hyperemic
spots ,erosives which can be changed to ulcers.
As the result there is oral mucosal discharges which has high
viscosity.
There may be ulcers on mucous membrane of nasal cavity and
vagina; and on muzzle.
After some days the affected animals(calves) develops diarrhea and
the faces is mixed with mucous or blood and has bad smelling.
The animal is dehydrated
In interdigital space of cloves, you may observe ulcers and erosion.
Diarrhea may continue upto 4 weeks and the animal die at
dehydration
Abortion in pregnant cows
In Subacute form of BVD, you may observe
Hyperthermia upto 40 0C for the short time
Depression and anorexia
Hyperemia in visible mucous membranes 1075
 BVD Cont’d---
Salivation
Nasal discharge and coughing
Atony of GIT
In some cases, there may be diarrhoea for short time
Subacute form of BVD ends with self recovery but affected
animals become carriers of the virus.
Chronic form of BVD often occurs at the end of every epidemics.
Clinical signs develop slowly even upto 6 months
The main clinical signs are
Emaciation
Constant or intermittent diarrhoea
Ulcers with necrotic mass on the mucous membrane of oral cavity
Prognosis rarely bad.
Latent form of BVD can be known only by serological method of
diagnose.
It occurs between the two epidemics of BVD.
This form of BVD is dangerous because the virus exist as persistent
1076
 BVD Cont’d---
and cause persistent infection(PI) and the affected animal becomes a
source of infection for long period of time.
Necropsy findings
The main necropsy findings that can be observed on animals died
at BVD are
Dehydration
Ulcers and erosion on mucous membrane of GIT( they are clearly
visible on gums, on soft and hard palate; on mucous membrane of
abomasum)
 The mucous membrane of abomasum and small intestine are
inflamed with haemorrhagiae.
Mesenterial lymph nodes are also inflamed.
Liver and kidney may be enlarged and have haemorrhagiae under
their capsule
Petechial haemorrhagiae may be found on epicard and endocard.
Ulcers and erosions may be found on nasal cavity, on mucous
membrane of vagina and on interdigital spaces of hoof. 1077
 BVD Cont’d---
Diagnosis
Diagnose of BVD can be done based on
Epidemiology
Clinical signs
Necropsy findings
These methods of diagnoses give tentative diagnose of the disease
Definite diagnose of the disease is given based on the result of
laboratory diagnose.
Appropriate specimens are
Mucous membrane of nasal cavity and intestine
Parenchymatous organs like spleen, kidneys and liver
Mesenterial lymph nodes
Virus isolation is done on primary cell culture of testicle of bulls,
spleen and kidney of embryo cattle.
The isolated virus is identified by using
Virus neutralization(VN)
RID(Reaction immunodiffusion ) 1078
 BVD Cont’d---
CFT(Complement Fixation Test)
FAT (Fluorescence Antibody test)
The disease can be diagnosed retrospectively using paired serum
samples using VN and CFT.
The diagnose is considered as positive if titer of antibody in the
second serum is 4 times that of the first.
It is also possible to diagnose BVD using antigen detection.
The BVDV antigen can be identified rapidly in tissue samples
using immuno histochemical methods such as
immunofluorescence and immunoenzyme staining in sections of
frozen tissue.
Skin biopsy by using immunohistochemical (IHC) staining for
BVDV in formalin fixed, paraffin embedded skin biopsies is an
effective method for the diagnosis of persistently-infected cattle.
The technique is easy, accurate, and less expensive antemortem
diagnostic test for the detection of PI animals compared to virus
isolation. 1079
 BVD Cont’d---
Using of laboratory tests in the herd can be used
1. Acute infections
 2. Persistently infected animals
The diagnosis of acute infections must be done as early as 3 - 8
days after infection.
A whole-blood sample is the best sample for virus isolation to
identify acutely infected animals.
In herd outbreaks, blood samples from normal animals should also
be submitted.
For serology, paired acute and convalescent samples collected 21 -
30 days apart are required to identify a four-fold increase in serum
antibody titers.
In abortions, the dam may have already seroconverted before the
abortion
So that there may be no seroconversion between acute and
convalescent sera
1080
 BVD Cont’d---
In persistently infected animals in a herd can be identified using any
or a combination of the following testing procedures:
 1. Collect whole blood from all animals in the herd including calves.
An RT – PCR can be done on every sample which is very expensive
but highly sensitive method.
 2. Collect serum on all animals over 3 months of age and detect the
presence of antibody using ELISA.
3. Collect skin biopsies (ear notches) from all animals in the herd
including calves. The tests of choice are the IHC on formalin-fixed
tissues or antigen capture ELISA using fresh samples.
Differential diagnose
BVD must be differentiated at many diseases of cattle those have
similar clinical features.
1. From diseases with oral lesions and diarrhea
 Rinder pest
 MCF
 Infectious Rhino Tracheitis(IRT 1081
 BVD Cont’d---
2. Diseases with oral lesions and no diarrhea
 FMD
 Vesicular stomatitis
 Blue tongue
 Bovine papular stomatitis
3. Diseases with diarrhea and no oral lesions
 Paratuberculosis
 Salmonellosis
 Emeriosis
 Intoxication with arsenic
 Ostertagiasis
 Carbohydrate engorgement
Treatment
There is no any specific effective method to treat animals from
BVD
However, hyperimmune serum and serum from recovered animals
were used but do not have high efficacy. 1082
 BVD Cont’d---
So using of supportive and symptomatic treatment may help to
reduce the mortality of animals.
Some of supportive treatments are
Intravenous administration of electrolyte and glucose to combat
dehydration, electrolyte and acid base imbalance.
 Cleaning and washing of the mucous membrane with effective
antiseptics like neuro furacilin.
Administration of Enemas using 1%Cuso4
Administration of cardiac preparates
 To control secondary bacterial complication we can use broad
spectrum antibiotics and sulfa drugs.
Preventation and control measures
The ultimate goal of BVDV prevention and control measures is to
eliminate the potential for the birth of calves persistently infected
with the virus.
So that a combination of biosecurity, vaccination and
biocontainment strategies are necessary to control and prevent 1083
 BVD Cont’d---
BVDV infection and its consequences in a herd and country.
Biosecurity is the action taken to prevent the introduction of a
disease agent into a herd or region.
Biocontainment includes the strategies to control an already existing
disease in a herd or region.
The successful control and prevention of the bovine virus diarrhea-
mucosal disease complex depends on
1. Identification and elimination of PI animals from the herd
2. Prevention of introduction of infection into the herd (biosecurity)
3. Immunization programs and biocontainment
4. Eradication of the virus from herds
1. Identification and elimination of PI animals from the herd
Identification and elimination of PI cattle is an essential component
of a control program in an infected herd.
Elimination of such animals, also known as “clearance of
infection”.
1084
 BVD Cont’d---
2. Prevention of introduction of infection into herd (biosecurity)
After identification and elimination of the PI animals, the new
virus-free status of the herd should be maintained by a program of
testing of all introduced animals for freedom from infection.
In many cases introductions can be guaranteed, as far as is
reasonably possible, to be free of infection by selecting animals
which have convincing titers of serum antibody or are negative.
3. Immunization programs and biocontainment
Some of specific preventation methods of BVD are
Passive immunization
Active immunization
Some of the passive immunization methods are
Specific immunoglobulin
Hyperimmune serum
Serum from recovered animals
Serum for preventation and control of BVD can be administered
Aerosol(2 -5 ml/ m3) 1085
 BVD Cont’d---
Intranasal (2 – 4 ml for each)
Subcutaneously(1 ml/ kg BW).
Active immunization can be done by using
Live attenuated for feed lot cattle
Killed vaccine for dairy farms( reproductive animals)
Do not use live vaccine for reproductive cattle when they are
pregnant.
Because calves obtained from vaccinated pregnant dams can be
infected and may have persistent infection.
Regularly mechanically clean and disinfect the animal house.
4. Eradication of the virus from herds
If an outbreaks occurs on the farm, sick animals must isolated and
treated animals with specific immunoglobulin or hyper immune
serum or serum from recovered animals.
The animal houses must be regularly cleaned and disinfected by
using appropriate disinfectants.
All animals must be tested for BVD and those which give positive
test result must be sold for meat.
1086
In this section we give attention on
3.1. Lumpy Skin Disease(LSD)
3.2. Sheep pox and Goat pox
3.3. Contagious Ecthyma( Contagious pustular dermatitis, orf)
3.4. Papillomatosis

1087
 Lumpy skin disease(LSD) is an acute to chronic viral disease of
cattle which is characterized by
Fever
Followed by the development of nodular lesions in the skin that
subsequently under go necrosis
General lymphadenitis and edema of limbs
Results in ulceration of lesions and development of secondary
bacterial infection.
LSD is also referred to as
Pseudourticarial disease
Etiology
LSD is caused by DNA virus belongs to
Family Poxviridae
Subfamily : - Chrodopoxvirinae
Genus : - Capripoxvirus.
Carpipox virus is DNA virus and it is the largest virus which has
1088
brick shape with complex structure.
 LSD Cont’d---
Antigenically, it is closely related to sheep and goat pox viruses.
These viruses cannot be differentiated using routine serological
testing.
Epidemiology
It was first recognised in Zambia in 1929.
This disease is widely prevalent only in Africa.
LSD affects cattle of both Bos taurus and Bos indicus.
Young animals are more susceptible than adults.
Occurrence of LSD is mostly related with wet and humid season.
So that epidemics of LSD occurs mostly in rainy season.

1089
 LSD cont’d---
Morbidity in susceptible herds may be as high as 100%.
But mortality is rarely more than 1 -2%.
The economic importance of LSD is prolonged convalescence and
in this respect, it is similar to FMD.
Source of infection
Clinically sick animals
Transmission
Transmission of LSD virus is primarily by biting insects.
Most cases result from a transmission by an arthropod vector.
Particularly mosquitoes under genus of Culex and Aedes.
Flies under genus of Stomoxys, Biomyia, Culicoides, Glossina and
even Musca .
Direct contact is a minor way of transmission.
However, LSD virus can be present in
Cutaneous lesions
Saliva and nasal discharge
Milk and semen 1090
 LSD cont’d---
Very rarely cattle can be infected by LSD virus with drinking
water.
However, ingestion is not a common route.
NB. No carrier state in LSD
Risk factors
The risk factors in LSD are
1. Animal risk factors
2. Pathogen risk factor
3. Environmental risk factor
1. Animal risk factors
All ages and breeds of cattle are susceptible to LSD except animals
recently recovered from an attack.
Because there is a solid immunity lasting for about 3 months.
In outbreaks, very young calves, lactating and malnourished cattle
develop more severe clinical disease than others.
Wildlife species are not affected in natural outbreaks although there
is concern that they might be reservoir hosts. 1091
 LSD cont’d---
There is only one report of the natural occurrence of LSD in a
species other than cattle.
This species is in water buffalo (Bubalis) and giraffe but no further
such cases are recorded.
2. Pathogen factors
Capri pox viruses are generally resistant to drying, survive freezing
and thawing.
Resistance to heat is variable.
But most are inactivated at temperatures above 60°C.
3. Environmental risk factor
Outbreaks tend to follow waterways and extensive epizootics are
associated with
High rainfall and
 Concomitant high levels of insect activity
That is why LSD outbreaks has a peak in the late winter and early
autumn in Ethiopia.
1092
 LSD cont’d---
Economic importance
The mortality rate is low but the economic loss is high.
The economic loss due to LSD comprises
Severe loss of milk production
The occurrence of secondary mastitis predisposed by the
development of lesions on the teats.
Damage to hides
Loss of body condition during the course of the disease
Loss of fertility in affected bulls
Abortion in course of the disease.
Lumpy skin disease is considered to be one of the high risk diseases
for spread out of Africa .
LSD may have the potential to spread out side African continents
through importation of wild animals( giraffe) to zoos.
It could establish new foci of infection if suitable insect vectors are
available.
1093
 LSD cont’d ---
Pathogenesis
In the generalized disease there is viremia accompanied by a febrile
reaction .
Localization in the skin occurs with development of inflammatory
nodules.
In the experimental disease following ID inoculation, local lesions
can develop at the site of challenge without viremia and
generalization of the infection.
Clinical findings
Incubation period of LSD is 2 - 4 weeks is common in field
outbreak.
In severe cases, there is
Fever lasting about a week
 Followed by lacrimation ,nasal discharge and salivation
 Lameness
Then multiple nodules appear suddenly a week later.
The first nodules usually appearing in the perineum. 1094
Lacrimation and nasal discharge during LSD

1095
1096
1097
 Rabies cont’d ---
Nodules are round and firm and vary from 1 to 4 cm in diameter.
They are flattened and the hair on them stands on end.
They vary in number from a few to hundreds and are intradermal.
In most cases, they are confined to the skin area.
Other manifestations that may be observed in severe cases include
Lesions in the nostrils and on the turbinate causing mucopurulent
nasal discharge, respiratory obstruction and snoring.
Plaques and later ulcers in the mouth causing salivation.
 Nodules on the conjunctiva causing severe lacrimation
 Nodules on the prepuce or vulva and spreading to nearby mucosal
surfaces.
The limbs may become grossly distended with edema fluid.
In most cases the nodules disappear rapidly.
But they may persist as hard lumps or become moist, necrotic and
slough.
Since drainage of lymph is disturbed the affected area become
1098
Lumpy skin disease. Various sized cutaneous nodules in a
severe case of lumpy skin disease.

1099
1100
1101
1102
 Rabies cont’d ---
enlarged and cause local edema.
When sloughing of the yellow center of nodules occurs there is
often exposure of underlying tissues, e.g. testicles or tendons.
Lesions where skin is lost may remain visible for long periods, and
where lesions have coalesced, large areas of raw tissue may be
exposed.
The skin lesions provide an excellent point of entry for screw
worms.
Pneumonia is a common sequel in cases where lesions occur in the
respiratory tract.
A convalescence of 4 - 12 weeks is usual.
Pregnant cows may abort.
Necropsy findings
Characteristic skin nodules are visible externally.
Similar nodules are present in the mouth, pharynx, trachea, skeletal
muscle, bronchi and stomachs and there may be accompanying
pneumonia. 1103
 LSD cont’d ---
The superficial lymph nodes are usually enlarged.
Respiratory distress and death are often the result of respiratory
obstruction by
 The necrotic ulcers and surrounding inflammation in the upper
respiratory tract and/or concurrent aspiration pneumonia.
Histologically, a widespread vasculitis reflects the viral tropism for
endothelial cells.
Intracytoplasmic viral inclusion bodies may be seen in a variety of
cells types.
Diagnosis
As usual diagnose of LSD is done based on
Epidemiology
Clinical signs
Necropsy findings
Laboratory diagnose
When you are considering the epidemiology of the disease
consider the basic information like 1104
 LSD cont’d ---
Species of animals that are affected( only cattle are always affected)
Seasonal occurs of disease( very often at the end of winter and
beginning of autumn season)
Morbidity almost 80 – 100% during LSD epidemics.
In LSD enzootic area the morbidity may reach 80%.
Case fatality not more than 2%.
Disease association with increased number of insects( mechanical
vectors)
When you consider the clinical signs, look the typical signs of LSD
like
Multiple skin nodules
Nodules around nostrils, turbinate, mouth, vulva and prepuce that
can be disappear or persist as hard lumps or become moist, necrotic
and slough.
Edema of leg
When you consider the necropsy findings, look the nodules in skin,
mouth, nostrils, vulva and prepuce. 1105
 LSD cont’d ---
Enlargement of superficial lymph nodes.
Edema of affected legs.
Histologically, a widespread vasculitis reflects the viral tropism for
endothelial cells
The confirmatory test is done in laboratory by antigen detection,
isolation and identification the virus.
Specimens of choices for virus isolation and identification are
Lymph node and skin lesion
NB. Parallel collect formalin-fixed lesions from skin, alimentary
and respiratory tissue, lymph nodes for histopathology.
Diagnosis is most commonly made by electron microscopic
demonstration of typical Carpipox virions in
Full thickness skin biopsies or
Scabs coupled with the clinical findings of a generalized nodular
skin disease with enlarged superficial lymph nodes.
Biopsy of lesions reveals a granulomatous reaction in the dermis and
hypodermis. 1106
 LSD cont’d ---
In the earlier acute stages, there are intracellular, eosionphilic
inclusion bodies.
If specimens are collected early in the course of the disease before
the development of neutralizing antibodies , the antigen of Carpipox
virus can be detected by
Direct FAT
ELISA
Molecular technique like PCR
LSD virus can be isolated by using cell culture.
Identification of the isolated virus can be done using Virus
Neutralization Test(VNT) using specific monoclonal antibody.
The past infection of the herd with LSD virus can be known by
using serological tests like
Indirect FAT
ELISA
AGID
However AGID is not commonly used because of its low specificity 1107
 LSD cont’d ---
and give false positive reaction as it cross reacts with antibodies of
bovine papular stomatitis virus and pseudo cowpox virus.
Differential diagnosis
LSD must be differentiated from
Pseudo – lumpy skin disease
Streptothricosis
Allergic dermatitis
Pseudo-lumpy skin disease (also known as Allerton virus which is
the general infection of cattle associated with bovine herpesvirus-
2.
The disease common in South Africa.
In pseudo – lumpy skin disease involves only the superficial layers
of skin and have circular structure and are smaller ( up to 2 cm in
diameter).
In contrast to the lesions of lumpy skin disease which are often
deep enough to expose underlying tissues.
1108
 LSD cont’d ---
Treatment
No specific treatment is available.
But prevention of secondary infection is essential.
So that it is possible to administer broad spectrum antibiotics or
sulfonamides.
Prevention and control measures
Lumpy skin disease moves into new territory principally by means
of
Movement of infected cattle and/or
Possibly by wind-borne vectors
In the new territory further spread is accepted as being via an
insect vector.
Control of cattle movement from uninfected to infected territory
is an important control measure.
Further control can only be by vaccination.
Nowadays the following vaccines are available in the world
1. A safe and effective vaccine has been produced by 60 passages 1109
 LSD cont’d ---
of the virus through lamb kidney culture.
It is administered to all animals over 6 months of age.
2. A freeze-dried, living attenuated virus vaccine is also available.
 3. The most commonly vaccine is vaccination of cattle with sheep
pox virus which is also attenuated by passage through tissue
culture.
It is effective in preventing infection with the lumpy skin disease
virus and is currently used in most African countries.
NB. In Ethiopia the vaccines that are used to control LSD are
Lyophilized live freeze dried Carpipox virus strain with TCID50
per dose.
It is available in 20 ml of vial of 100 dose.
Reconstitute it with 100 ml of sterile saline water.
Dose and route of administration : - 1ml S/C
Immunity develops after eight days and lasts for two years.
It is also possible to use sheep and goat pox vaccine but reduce the
dose by half .
1110
Inject 0.5 ml of sheep pox vaccine S/C to cattle.
 They are the most serious of all pox diseases of domestic animals
characterized by
Fever
Edema of the eyelids
Mucous discharge from nose
Appearing of cutaneous nodules all over the body, most obvious in
the area of skin where the wool or hair is short.
Etiology
The virus is DNA virus found under
The family of Poxviridae
Subfamily Chrodopoxvirinae
Genus Capripoxvirus
Sheeppox and Goatpox are host specific.
But some strains affect both species.
That is why, three viruses are named on the basis of their host
specificity in natural outbreaks.
1111
 Pox cont’d---
1.Sheeppox virus
2.Goatpox virus
3. Kenya Sheep and Goatpox virus
Sheeppox virus and Goatpox virus are mainly highly host specific
in natural infections.
But exceptions exist, Kenya sheep and Goatpox virus and Yemen
and Oman sheep isolates infect both sheep and goats.
All are members of the genus Capripoxvirus and are closely
related on the basis of genome mapping.
Epidemiology
Sheeppox and goatpox are prevalent in Africa except South Africa.
It is also prevalent in most developing countries including the
Indian subcontinent, Middle Eastern countries and China.
However, sporadic outbreaks occur in southern Europe and
elsewhere.
The capripoxvirus infections of small ruminants are the most
serious of all the pox diseases in animals. 1112
 Pox cont’d---
In susceptible flocks and herds morbidity is 75 - 100% with
outbreaks often causing death with average of 50%.
Mortality in lambs may reach 100%.
Source of infection
Sick animals
Carriers with latent infection
Transmission
Sheeppox and goatpox are highly contagious diseases.
The virus enters via
 The respiratory tract and transmission commonly is by aerosol
infection associated with close contact with infected animals.
 Contaminated materials and through skin abrasions produced
iatrogenically or by insects.
The virus is present in nasal and oral secretions for several weeks
after infection and can live in scabs that have fallen off the animal
for several months.
Spread can also occur from contact with contaminated materials1113 and
 Pox cont’d---
through skin abrasions produced iatrogenically or by insects.
Capripox has been shown to spread via the bites of Stomoxys
calcitrans and the tsetse fly.
Risk factors
The risk factors are
1. Animal risk factors
2. Pathogen risk factors
1. Animal risk factors
It is known both sheep pox and goat pox affect sheep and goats of
all ages, breeds and sex groups.
But young and old animals and lactating females are more severely
affected.
In areas where sheep pox is enzootic, imported breeds such as
Merinos or some European breeds may show greater susceptibility
than the native stock.
2. Pathogen risk factors
The virus is resistant to drying and survives freezing and thawing.
1114
 Pox cont’d---
It is sensitive to extremes of PH and 1 % formalin.
Sensitivity to heat and most strains are inactivated at 60°C for 60
min.
Isolates from most regions are host specific
But isolates from Kenya, Yemen and Oman naturally infect both
goats and sheep.
Scabs shed by infected animals remain infective for several months.
Economic importance
The economic importance of sheep pox and goat pox comprises
Loss is from direct mortality
Loss due to abortions and mastitis
Loss from down grading and skin condemnation
Loss from poor quality of wool
Loss of exports.
In ewes severe losses may occur if the udder is invaded because of
the secondary occurrence of acute mastitis.
NB. Sheep pox and Goat pos have no zoonotic implication. 1115
 Pox cont’d---
Pathogenesis
During an initial viremia, the virus is deposited in most tissues,
including the skin.
The virus is present in greatest quantities between the 7th and 14th
day after inoculation.
Circulating antibody limits spread of infection, but does not prevent
replication of virus at the site of inoculation.
Since the virus is epitheliotropic, it will infest the epithelium tissues
of the organism.
Generally, pox infections follow five stages while producing the
lesions.
Development of pox lesions
There are 5 stages in the development of pox infection.
They are
1. Roseola stage
2. Popular stage
3.Vesicular stage 1116
 Pox cont’d---
4. Pustular stage
5. Stage of desquamation
1.Roseola stage
Skin lesions typically begin with small red spots with in three days
of infection which is followed by papules.
The affected animals are febrile at this stage.
2. Papules
Nodular skin lesions that are developed from roseola stage (red
spots) those are hard during palpation.
Papules develop after 3 days of roseola stage.
The affected animals are also febrile at this stage.
3. Vesicular stage
Papules with in 5 – 6 days are changed to vesicles.
Vesicles are characterized by having gray – yellowish exudate.
At this stage fever reduces.
4. Pustular stage
Pustular stage develops after 3 days of vesicular stage. 1117
Roseola stage of sheep pox and goat pox

1118
Papules stage of sheep pox and goat pox

1119
1120
Vesicular stage of sheep pox and goat pox

1121
1122
1123
5 th stage also called stage of desquamation.

1124
1125
 Pox cont’d---
At this stage a gray – yellowish exudate contents of vesicles is
changed into high viscous fluid called pus.
Pustules with pus may
Burst or
Become desiccated
The larger ones may leave a pock mark which can be a deep lesion
with permanent scarring.
Pock meaning having holes or depressions in the surface.
This stage of pox development is called the 5 th stage also called
stage of desquamation.
Clinical findings
The clinical findings during sheep pox and goat pox infection
depends what type of Capripoxvirus affect the susceptible animals.
Depending the above fact, there are different pox infections with
different clinical findings in small ruminants.
These are
1.Sheep pox in sheep 1126
 Pox cont’d---
2.Goat pox in sheep
3.Goat pox in goats
1.Sheep pox in sheep
Incubation period ranges from 2 to 14 days.
The disease can occur in two forms:.
A. Malignant form
B. Benign form
A. The malignant form
This form of sheep pox in sheep is more common in lambs and is
characterized by
Marked depression and prostration
Edema of eyelids
 A high fever(41 – 42 0C) and discharges (serous or pus)from the eye
and nose.
 Difficulty in breathing due to accumulation of exudate in trachea
and bronchi.
Affected lambs may die during this stage before typical pox lesions
1127
Sheep pox in lambs

1128
1129
 Pox cont’d---
develop.
However, if the lambs do not die
The pox lesions often appear after 4 days around
Head
Lips
Nostrils
Under tail
Orbit of eye
In the inner thigh of animals
Rarely ventral part of abdominal cavity, udder and scrotum.
Them pox lesions commence as papules then nodular, vesicular,
pustular and finally scabs.
During sheep pox in sheep, pox lesions are seen mainly on
Un wooled skin and
 The buccal, respiratory, digestive and urogenital tract mucosae
B. The benign form
This form of sheep pox in sheep is common in adults. 1130
1131
 Pox cont’d---
And only skin lesions occur particularly under the tail
There is no systemic infection
Skin lesions in sheep may cause intense irritation and/or pain
leading to self mutilation
2. Goat pox in sheep
Goat pox in sheep is more severe than sheep pox and lesions occur
On the lips and oral mucosae
On the teats and udder
3. Goat pox in goats
It is very similar clinically to sheep pox in sheep.
It has the morbidity of 90% and mortality rate of 4% in susceptible
goat flocks.
Young kinds suffer from systemic disease
In adult goats the disease has mild form.
Necropsy findings
In addition of skin lesions that are visible externally on un wooled
parts of skin, there are necropsy findings in internal organs in 1132
1133
1134
1135
 Pox cont’d---
malignant form.
In the malignant form, pox lesions extend into the mouth, pharynx,
larynx, and vagina with lymphadenopathy and a hemorrhagic spleen.
Lesions may also appear in the trachea and lung with severe
manifesting as lentil sized white pox nodules to a consolidating
necrotizing pneumonia.
Lesions occasionally reach into the abomasum and are accompanied
by a hemorrhagic enteritis.
Diagnosis
Diagnose of sheep and goat pox can be made based on
Epidemiology of the disease
Clinical signs
Necropsy findings
Laboratory diagnosis
However, diagnosis is based on typical clinical signs combined with
laboratory confirmation of the presence of the virus or antigen.
The virus can be detected by using electron microscopy and shows1136
Sheep pox. Reddish to whitish nodules in the lungs.

1137
 Pox cont’d---
large numbers of characteristic 'sheep pox cells' containing inclusion
bodies and typical capripox virions can be seen in biopsies of the
skin.
The virus can be isolated in tissue culture.
But virus isolation as a method of rapid diagnosis is limited
Because some strains need several blind passages for virus to show
cytopathic effects(CPE).
The sheep pox virus antigen can be detected by
Direct fluorescent antibody test using edema fluid
 Agar gel immuno Diffusion(AGID) using biopsies of lymph nodes
using specific immune sera.
ELISA can be used to detect sheep and goat pox antigens.
The past infection of the flock can be known by using serological
tests.
The antigen commonly used is a' soluble antigen fraction' which 1138is
 Pox cont’d---
non infectious, has replaced infectious virus for serological tests
which avoids the risk of accidental spread of the virus from
diagnostic laboratories.
The most common serological tests that are used in most
laboratories are
AGID
Indirect FAT
Virus Neutralization Test(VNT)
ELISA
But these serological tests are not widely used
Because of their low specificity and sensitivity as they are
confounded due to cross reaction with orf (contagious pustular
dermatitis virus).
Differential diagnosis
Sheep and goat pox must be differentiated from
Contagious pustular dermatitis virus( orf)
Blue tongue 1139
 Pox cont’d---
Treatment
No specific treatment is advised.
But palliative treatment may be necessary in severely affected
animals.
Prevention and control measures
Prevention and control measures of sheep pox and goat pox depends
on epidemiology the disease in a given countries or regions.
1. Control in free countries or regions
It is necessary to prohibit the importation of sheep and goat from
infected areas.
And in a case if infection is introduced do
Ring vaccination
Destruction of affected flocks and quarantine of infected premises
should be instituted .
2. Control in enzootic areas
Vaccination is the basis for control sheep pox and goat pox.
It is known that natural infection with one capripox strain imparts
1140
 Pox cont’d---
immunity to all capripox infections.
So that vaccination with a single capri pox vaccine will give
protection across all species and against all capripox infections.
A large variety of commercial vaccines is now available.
The vaccines can be
Killed virus vaccines that elicit at best, temporary protection
Live attenuated vaccines appear to give excellent protection for
periods greater than 1 year.
You know colostral antibody interferes with response to vaccination
before 6 months of age.
So that it is not advisable to vaccinate lamps and goat kids before 6
months of age.
In Ethiopia, sheep pox and goat pox vaccine is available.
It is lyophilized live freeze died Carpipox virus strain in vial of 20 ml
of 100 doses.
Reconstitute the vaccine with 100 ml sterile saline water.
1141
 Pox cont’d---
Dose and route of administration : - 1 ml S/C in the inner thigh of
sheep and goat.
Immunity develops after 8 days and lasts for 2 years.
NB. Vaccination in the face of outbreak is unlikely to prevent deaths
during the subsequent 2 weeks because if needle hygiene is poor, may
facilitate the spread of the disease.

1142
 It has different names as
Orf
 Scabby mouth
Sore mouth
Contagious ecthyma is a viral disease of mainly sheep and goat
characterized by formation of nodules, vesicles, pustules and
scabs on mucous membrane oral cavity, lips, distal part of limbs,
udder, reproductive organs and rarely on other parts of affected
animals.
Etiology
The virus is DNA virus belongs
Family Poxviridae
Genus Parapoxvirus
In addition to the orf virus (Parapox ovis) the genus includes the
Viruses of bovine papular stomatitis:- Parapoxbovis 1
Pseudocowpox : - Parapoxvirusbovis 2 1143
 Orf Cont’d---
 Parapoxvirus of deer.
The virus can be cultivated in chick embryo at the age of 9 – 12
days and mild lesions develop on the chorioallantois membrane.
The orf virus withstands drying and is capable of surviving at room
temperature for at least 15 years.
Restriction endonuclease analysis of DNA shows considerable
heterogeneity between different field isolates.
Epidemiology
In natural condition contagious ecthyma affects goats and sheep of
all breeds, sex and age.
Lambs and goat kids at the age of 4 days up to 10 months
susceptible.
Orf occurs most commonly in lambs 3 - 6 months of age at pasture
time.
Although outbreaks occur at any time but they are most common in
dry conditions when the sheep are at pasture, or in penned sheep
being fed from feed troughs. 1144
 Orf cont’d---
The disease also occurs in humans working among infected sheep.
So orf has zoonotic implication.
In abattoir workers it is commonest in those handling wool and
skins.
Source of infection
Source of infection during orf can be
Sick animals
Recovered animals
Means of transmission
The virus can infect the pasture and animal feeds, feed trough etc
that is expelled from the organism together with
Secrete of pustule and vesicle
Scabs and discharges from oral cavity.
The route of infection is mechanically damaged skin, mucousa
membrane, distal parts of limbs, lips and reproductive organs.
Infection can be from
 Environmental persistence of the virus or 1145
 Orf Cont’d---
Infected sheep
Spread in a flock is very rapid and occurs by contact with other
affected animals or by contact with contaminated inanimate objects.
It has been assumed that natural infections on pasture are the result
of invasion of the virus after skin damage induced by prickly plants
or stubble.
Morbidity rates approaching 100% and case fatality rates from 5 to
15% .
The deaths that occur are due to the extension of lesions in the
respiratory tract.
Risk factors
The primary risk factors are
Pathogen risk factor(the presence of the virus in the environment)
The immune status of the sheep and goat
The presence of predisposing factors
Economic importance
The disease produces a minor set back as it has low mortality rate.
1146
 Orf Cont’d---
However when the orf disease affects young sucking lambs with
associated lesions on the teats and udders of their ewes.
Loss from lamb mortality and secondary mastitis in these
circumstances can be significant.
Pathogenesis
Damage to the skin is essential for the establishment of orf
infection and the development of typical lesions.
After infection of the orf virus replicate in epithelium cells of skin
cloves, lips and reproductive organs specially in vulva and prepuce.
This causes proliferation and degenerative process in affected
epithelial cells and form nodules with red colour.
And there is formation of cytoplasmic inclusions and exudation.
As the result the nodules are changed to vesicles then to pustules.
Lastly the thick crust forms which can fail to the ground.
What it makes different from sheep pox and goat pox is it never
forms scar in affected epithelium tissues.
In unfavorable condition the pustules may join together and are1147
 Orf Cont’d---
changed to erosion and may have generalized form.
Clinical findings
1. Orf in sheep
New lesions will develop during the first 10 days of infection.
Lesions develop initially as papules and then pustules.
Time progression from the initial lesions to the formation of scabs is
approximately 6 to 7 days.
The first lesions develop at the oral muco - cutaneous junction.
Usually at the oral commissures and are accompanied by swelling
of the lips.
From here they spread on to the muzzle and nostrils to a lesser
extent on to the buccal mucosa.
They may appear as discrete thick scabs 0.5 cm in diameter or
coalesce and be packed close together as a continuous plaque.
Affected lambs suffer a severe setback.
Because of restricted sucking and grazing.
Cases the scabs dry and fall off and recovery is complete in about
11483
 Orf Cont’d---
weeks.
Affected lambs sucking ewes may cause spread of the disease to the
udder.
Lesions on the teats predispose to mastitis and secondary infection
In rams, lesions on the scrotum may be accompanied by fluid
accumulation in the scrotal sac and associated temporary infertility.
A high incidence of infection can also occur where the dominant
lesions are on the feet, occurring around the coronary band.
Rarely, systemic invasion occurs and lesions appear on the coronets
and ears, around the anus and vulva or prepuce, and on the nasal and
buccal mucosae.
There is a severe systemic reaction, and extension down the
alimentary tract may lead to a severe gastroenteritis, and extension
down the trachea may be followed by bronchopneumonia.
A malignant form of the disease has also been observed in sheep.
It begins with an acute episode manifested by oral vesicles, and
extension of these lesions down the gastrointestinal tract, followed
1149
Sore mouth in Sheep

1150
1151
1152
1153
1154
1155
1156
1157
1158
1159
Orf in human being

1160
 Orf Cont’d---
later by granulomatous lesions and shedding of hooves.
2. Orf in goats
Almost it has similar clinical sign with orf in sheep.
Multifocal lesions occurred over the head, neck, thorax, and flanks
of each animal.
The lesions developed approximately 2 weeks after the animals
returned from a local summer.
The skin crusts gradually dried and fell off, leaving areas of
alopecia and depigmented skin.
Necropsy findings
In malignant cases there are irregularly shaped lesions with a
hyperemic border in the oral cavity and the upper respiratory tract.
These irregular shaped lesions rarely may involve in the mucosae
of esophagus, abomasum and small intestine.
Diagnosis
Orf as other viral diseases can be diagnosed by complex method.
Epidemiologically 1161
 Orf Cont’d---
Clinically
Necropsy findings
Laboratory
Samples for confirmation of diagnosis can be
For virology, the specimen of choices are vesicle fluid, scraping
from active lesions
Parallel collect for histopathology which is fixed with buffer
formalin.
Electron microscopic identification of the virus is quick and
generally reliable with multiple samples from an affected herd or
flock.
Virus can also be detected by PCR and restriction enzyme analysis
of viral DNA and gene sequencing.
Recovered animals have a high level of neutralizing antibodies in
their serum and this is detectable by a gel diffusion test and other
serological tests
But has little clinical value. 1162
 Orf Cont’d---
Differential diagnosis
Orf must be differentiated from many infectious and non –
infectious diseases of sheep and goat with similar clinical signs.
Mycotic dermatitis
Facial eczema
Papillomatosis (warts)
Bluetongue
Sheeppox
Foot and mouth disease
Treatment
There is no specific treatment.
Removal of the scabs and the application of ointments or astringent
lotions are practiced.
But delay healing in most cases.
The provision of soft palatable food is recommended.
Prevention and control
In the early stages of an outbreak, the affected animals should be1163
 Orf Cont’d---
isolated and the remainder vaccinated.
Vaccination is of little value when a large number of animals are
already affected.
Persistence of the disease in a pastured flock from year to year is
common.
In such circumstances the lambs should be vaccinated at 6 - 8 weeks
of age.
The vaccine is live attenuated vaccine.

1164
 It has also another name called warts.
Papillomatosis affects cattle, horses, sheep and goats.
It is cutaneous benign tumors induced by host-specific
papillomaviruses.
The viruses infect epithelial cells causing hyper proliferative lesions
that are benign, self-limiting.
In most case papillomatosis in animals are spontaneously regress.
Differentiation of types is based on
The histological features of the lesion and
 DNA identification by hybridization or PCR
There has been little research on the papilloma virus of horses or
small ruminants.
In cattle six types have been identified and include
Bovine papilloma virus - 1(BPV – 1)
Bovine papilloma virus - 2(BPV – 2)
Bovine papilloma virus - 3(BPV – 3)
1165
 Bovine papilloma virus - 4(BPV – 4)
 Bovine papilloma virus - 5(BPV – 5)
 Bovine papilloma virus - 6(BPV – 6)
 Bovine papilloma virus BPV– l, BPV - 2 and BPV - 5cause
fibropapilloma
 BPV - 3, BPV - 4 and BPV - 6 cause true epithelial papillomas.
Cattle types show some site predilection or site specificity.
 BPV- 1 causes frond fibropapillomas of teat skin and penile
fibropapilloma.
BPV- 1 and BPV-2 causes fibropapilloma of the skin of the
anteroventral part of the body including the forehead, neck and
back. It causes the common cutaneous wart in cattle.
 BPV-2 may also cause cauliflower-like fibropapillomas of the ano -
genital and ventral abdominal skin
 BPV-2cause cancer in bladder which is associated with the
ingestion of bracken fern (Pteridium spp)
1166
 BPV-3 causes cutaneous papilloma
 BPV-4 causes papilloma of the esophagus, esophageal groove,
forestomachs and small intestine. It has capable of becoming
malignant, particularly in animals fed bracken fern.
Although a single BPV type is detected in an individual papilloma,
a single animal may have papillomas at different sites associated
with different BPV types.
Papilloma of cattle that have regional distribution and may have
separate antigenic identity are
 Oral papillomas, mostly in adult cattle these are caused probably
by BPV-4.
Papilloma of the larynx in steers
 Papillomavirus has been observed in squamous cell carcinoma of
bovine eyes.
Other skin lesions in which papillomavirus plays an etiological
role are
Equine sarcoid which associated with BPV-1 and BPV-2
Ear cancer of sheep 1167
 Palp cont’d ---
Epidemiology
Papillomatosis has an international occurrence in all animal
species.
The method of spread is by direct contact with infected animals,
infection gaining entry through cutaneous abrasions.
Virus can also persist on inanimate objects in livestock buildings
and infect animals.
Crops of warts sometimes occur around ear tags, at branding sites
or along scratches made by barbed wire rubbing against them.
An extensive outbreak of perianal warts is recorded in beef heifers,
the infection having been spread by rectal examination for
pregnancy
Risk factors
The commonest risk factors that exposed animals to papillomatosis
are
 Species
 Age 1168
 Papl. Cont’d---
 Immune status of the organism
All species of wild and domestic animals are susceptible to
infection
But the disease is common in cattle and horses.
Outbreaks have been recorded in sheep and goats but the disease is
uncommon in sheep.
It is also uncommon in pigs but if it occurs usually affecting the
genitalia of pigs
Cutaneous papillomas of the head and neck occur predominantly in
young animals of cattle.
The lack of susceptibility of adults to natural infection being
ascribed to immunity acquired by apparent or inapparent infection
when young.
The occurrence of cutaneous warts and their severity can be
influenced by factors that induce immunosuppression.
Latent infection has converted to clinical disease with the
administration of immunosuppressive agents like feeding of 1169
 Papl. Cont’d---
bracken fern
NB. Congenital infection is recorded in the foal and calf but it is
rare.
Economic importance
In purebred animals they may interfere with sales because of their
unsightly appearance.
Animals with extensive lesions may lose condition, and secondary
bacterial invasion of traumatized may cause concern.
Warts on the teats of dairy cows often cause interference with
milking.
In all species, the development of warts on the genitalia requires
immediate treatment.
Pathogenesis
Papiloma virus infects the basal keratinocytes, replicating its
genome in the differentiating spinous and granular layers causing
the excessive growth that is characteristic of wart formation.
The tumor contains epithelial and connective tissues and can be1170a
 Papl. Cont’d---
papilloma or a fibropapilloma.
Papiloma or fibropapilloma formation depends on the relative
proportions of epithelial and connective tissue present in the wart
Papillomas contain little connective tissue and fibropapillomas are
mostly fibrous tissue with very little epithelial tissue.
Clinical findings
It is known that warts are solid outgrowths of epidermis.
Structurally they can be
 Sessile
 Pedunculated
Clinically warts have different forms in different species of
animals.
I. Warts in cattle
Warts in cattle occur on almost any part of the body.
The most common papillomas occur in the skin of cattle under
2 years of age.
They are located most commonly on the head, especially around 1171
 Papl. Cont’d---
the eyes, on the neck and shoulders.
But they may spread to other parts of the body.
They vary in size from 1 cm upwards.
They have dry, horny, cauliflower-like appearance.
In most animals they regress spontaneously.
But the warts may persist for 5 - 6 months, and in some cases for as
long as 18 months, with serious loss of body condition.
By their anatomical location warts in cattle can be
1.Teat warts
2. Perianal warts
3. Genital warts
4. Alimentary tract warts
5. Urinary bladder warts
1.Teat warts
They may have
 Frond form which is filiform projections on teats.
Frond form teat warts appear to have been drawn out into an 1172
 Papl. Cont’d---
elongated shape of about 1 cm in length.
Other forms of warts on teats are a flat, round type.
They are usually multiple, always sessile and upto 2 cm in diameter.
The third form has an elongated structure appearing like a rice
grain.
Teat warts may regress during the dry period and recur with the next
lactation.
2. Perianal warts
Perianal warts are esthetically unattractive warts but they do not
appear to reduce activity or productivity.
3. Genital warts
Genital warts are commonly found on the vulva and penis
They make mating impracticable.
Because the lesions are of large size, friable and bleed easily.
They occur on the shaft or on the glands of the penis in young bulls.
They can be single or multiple.
They are pedunculated and they frequently regress spontaneously. 1173
 Papl. Cont’d---
4. Alimentary tract warts
Alimentary tract warts are rarely observed clinically in farm animals
But they are recorded in abattoir with a high incidence in some
anatomical sites.
Papillomas in cattle mostly occur on
The lateral and dorsal aspects of the tongue
The soft palate
Oropharynx
Esophagus, esophageal groove
 Rumen and reticulum
Papillomas occurring in the esophageal groove and in the reticulum
are a cause of persistent ruminal tympany.
5. Urinary bladder warts
It is less common manifestations of papillomatosis in cattle.
It causes no clinical signs but may predispose to enzootic
haematuria.
Urinary bladder warts is common in cattle being fed bracken fern.1174
 Papl. Cont’d---
Because bracken fern is an immunosuppressor and mutagens in
bracken fern cause neoplastic transformation of papilloma cells to
other organs.
For example BPV – 4 which cause papillomas in the upper
alimentary tract can be transformed to urinary bladder in cattle
being feed bracken fern.
II. Warts in goats
Papillomas most commonly occur on the face and ears .
But may occur on the skin generally, especially on un pigmented
skin.
Most completely regress, others regress and recur.
But occasional lesions may progress to carcinomas.
Papillomas that occur on the teats are persistent and may spread
through the herd.
Necropsy findings
Necropsy finding has importance when warts are located in internal
organs(alimentary and urinary warts). 1175
 Papl. Cont’d---
Diagnosis
Diagnose of external warts mostly depends on clinical findings.
However laboratory methods help us to confirm the disease.
There are no specific changes in the haemogram or blood
chemistry.
But cattle with papilloma have lower numbers of CD4 lymphocyte
subpopulations.
Biopsy of a lesion can be used as specimen.
Biopsy used to observe the histopathological changes and to
identify the virus.
Microscopically it is easy to differentiate the true papillomas from
fibropapillomas.
Because papillomas consist of a hyperplasic epidermis with scant
dermal tissue.
While in fibropapillomas the dermal component tends to be
predominate.
In laboratory the need to identify the specific virus in a crop of 1176
 Papl. Cont’d---
warts creates a requirement for serological and molecular
techniques.
The most common techniques are
An ELISA
 PCR on biopsy material or tissue scrapings
Differential diagnose
Warts in cattle must be differentiated from at atypical papilloma.
Atypical papilloma in cattle is associated with an unidentified type
of the papillomavirus.
However, it is easy to differentiate because
Atypical papillomas are characterized by an absence of dermal
fibroplasia but presents in wart.
All ages of animals can be affected and the lesions persist for long
periods.
 They are characteristically discrete, low, flat and circular and often
coalesce to form large masses.
The external fronds are much finer and more delicate than warts. 1177
 Papl. Cont’d---
Warts in horses must be differentiated from
Sarcoid(locally aggressive benign fibroblastic tumors of the skin)
Melanoma
Treatment
Warts can be removed by surgery or cryosurgery.
However, there is a tendency for spontaneous recovery.
So that surgical removal followed by vaccination with an
autogenous vaccine give a good result.
Since surgical intervention and even vaccination, in the early stages
of wart development may increase the size of residual warts and
prolong the course of the disease.
This indicates treatment of warts must not begin in the early
development of warts.
Vaccination in cattle is done by autogenous vaccines prepared from
wart tissues of the affected animal
Commercially the vaccine are available but may be less efficacious.
1178
 Papl. Cont’d---
However, an autogenous vaccine prepared for a specific problem
has the advantage.
Because it includes the local virus types.
The vaccine is prepared from homogenized wart tissue that is
filtered and inactivated with formalin.
Because of the different BPV types, care is required in the selection
of the tissues.
In general terms they can be selected on tumor type, location, and
histological composition.
The alternative is to use many types of tissue in the vaccine.
The stage of development of wart is also important in preparation
of efficient vaccine.
Since papilloma virus is present in much greater concentration in
the epithelial tissue of older warts than young ones.
So that tissues those are used in preparation of vaccine must be
taken from older warts than young ones.
1179
 Papl. Cont’d---
The vaccine can be administered subcutaneously.
However, better results are claimed for Intradermal injection for
2 - 4 injections in 1 - 2 weeks apart are commonly recommended.
Preventation and control measures
Specific control procedures are usually not instituted or warranted.
Because of the unpredictable nature of the disease and its minor
economic importance.
However the following preventation and control measures may
reduce the occurrence of warts in animals.
Vaccination of animals with vaccine which has all serotypes of the
papillomavirus because they are very type-specific.
Avoidance of close contact between infected and uninfected animals
The use of communal equipment between affected and unaffected
animals should be avoided.

1180
In this portion we give attention on
4.1. Rift Valley Fever(RVF)
4.2. Nairobi sheep disease
4.3. Blue tongue

1181
 Rift valley fever is an acute viral vector born( insect transmitted)
disease of cattle, goat and sheep.
The disease is characterized by
Fever
Inappetance
Mucopurulent nasal discharge
Bloody diarrhea
Hepatitis in calves, lambs and goat kids
Abortion of pregnant animals
RVF affects human beings so that it is zoonotic.
RVFV cause flu- like illness, hepatitis, hemorrhagic fever and
retinitis in human being.
Etiology
Rift Valley Fever virus is a member of the family Bunyaviridae

1182
 RVF cont’d---
The family consists four genuses
Bunyavirus
Phlebovirus
Nairovirus
Hantavirus
All are RNA viruses in the family Bunyaviridae.
Rift valley fever is caused by Phlebovirus.
The first three genuses are arboviruses.
However, Hantavirus is not arbovirus.
Arbovirus means viruses those maintained in arthropod –
vertebrate – arthropod cycles.
But Hantavirus is non – arbovirus that means it has vertebrate –
vertebrate cycle( Man – Rodents) with out arthropod vectors.
Hantavirus causes disease to human beings transmitted from
rodents’ through their urine, faces and saliva.
The two common diseases in human beings those are caused by
Hantavirus are 1183
 RVF cont’d---
 Hantavirus hemorrhagic fever with renal syndrome(HFRS)
 Hantavirus pulmonary syndrome (HPS).
Rift valley fever is caused by Phlebovirus.
Genus Phlebovirus contains two serogroups and at least 50 viruses.
All of which are transmitted by sand flies or mosquitoes.
The genus contains important pathogen including Rift Valley Fever
virus and the sandy fly fever viruses.
Isolates from outbreaks in different countries are antigenically
similar and there is only minor genetic variation between isolates.
There is only one serotype of RVF virus.
However, there are differences in pathogenicity among strains of
RVF virus.
It is one of the most important veterinary pathogen in the world.
Because it has great potential to cause lethal epidemics disease in
sheep and cattle.
Far beyond its usual habitat in sub – Saharan Africa and great
potential cause severe human disease in such epidemics, it has 1184
 RVF cont’d---
potential to expand through out the world.
Epidemiology
Rift Valley Fever was initially reported in the Rift Valley in Kenya.
Epidemics in sheep, goats and cattle have been recognized in
southern and eastern African countries.
It was the time when intensive livestock husbandry was introduced.
In late 1997 and 1998 a major epidemic of RVF spread from
Somalia through Kenya into Tanzania.
The epidemics caused death of thousands of sheep, goats and
camels.
In 1998 there had been more than 90000 human cases and more than
500 death.
This epidemics considered the largest ever seen in eastern Africa.
It was blamed on exceptional rainfall as a result of an EL Nino
climate pattern.
In eastern, western and southern Africa, RVF virus survives in a
minimally evident endemic cycle for years. 1185
 RVF cont’d---
Then , when there is a period of exceptionally heavy rainfall, the
virus explodes in epidemics of great magnitude.
Although such epidemics had been studied for many years, it was
not known until 1980s the mechanism of this phenomenon.
It was found that the virus is transmitted transovarially among
flood water Aedes spp. of mosquitoes.
The virus survives for very long periods in mosquito eggs laid at the
edge of usually dry depressions called “dambos”.
Dambos are common throughout grassy plateau regions.
However, when the rains come and these dambos flood, the egg
hatch and infected mosquitoes emerge
And infect nearby wild and domestic animals.
This discovery involved one of the first successful application of
satellite remote sensing and geographic information system
technology.
Nowadays, it exists and occurs as epizootics throughout sub-
Saharan Africa with recent extensions into Egypt, Madagascar, 1186
small dambos in dry seasons

1187
Dambos after flooding

1188
 RVF cont’d---
Mauritania and most recently expansion to the Arabian Penunsula.
It has great potential for spread to other countries.
Source of infection
The virus is believed to be maintained through a cycle involving
mosquito vectors, wildlife and domestic livestock .
The virus is transmitted by transovarially transmission in certain
floodwater Aedes mosquitoes which have drought-resistant eggs
that survive for long time.
Epizootics occur in enzootic areas when wet and flood conditions
promote the expansion of the vector population in the presence of
susceptible livestock.
Ruminants are highly susceptible and serve as the main amplifying
host.
The vector is haematophages; they suck the peripheral blood
when a pronounced when short viremia occur in infected animals.
This facilitates the spread of the disease by biting insects.
Virus is also present in milk, feces and aborted fetuses. 1189
 RVF cont’d---
An unidentified wildlife reservoir host may also exist.
Method of transmission
A wide variety of mosquitoes from several genera have been
identified as vectors.
In addition to Aedes mosquitoes
Sand flies
Culicoides
They have also been identified as vectors but are not maintenance
hosts
Risk factors
The risk factors for occurrence of RVF are
1. Environment risk factors
2. Animal risk factors
1. Environment risk factors
The incidence of the disease varies with the size of the vector
population.
It is greatest in seasons of heavy rainfall which allows the vector
1190
population to increase and expand from permanent water sites to
Aedes mosquitoes

1191
Sand fly

1192
Culicoides

1193
 RVF cont’d---
breed in surface waters in normally dry areas.
Expanding irrigation schemes may also enlarge areas at risk.
2. Animal risk factors
There may be a high incidence of abortions and some deaths in
adult sheep and cattle.
Losses are due mainly to deaths in young lambs and calves
Mortality is higher in lambs than in calves.
Indigenous breeds may have inapparent infections.
Camels, domestic buffalo, monkeys, humans, mice, rats, ferrets, and
hamsters are susceptible to infection and goats moderately.
But pigs, rabbits, guinea pigs, and poultry are not susceptible to
RVF infection.
Trade animals are suspect as the source of infection to previously
free areas.
Pathogenesis
Hepatocytes are the primary site of viral replication in lambs and
calves. 1194
 RVF cont’d---
So that age is a determining factor in the progression and outcome of
infection.
In very young animals, hepatic lesions progress from degeneration
and necrosis of individual hepatocytes to extensive necrosis
throughout the liver resulting in hepatic insufficiency and failure.
In young animals, encephalomyelitis may also occur.
Clinical findings
Infected sheep develops
Fever
Inappetance
Mucopurululent nasal discharge
Bloody diarrhea
Under field conditions, 90 – 100% of pregnant ewes abort and called
abortion storm.
There is a mortality rate of 90% in lambs and 20 – 60% in adult
sheep.
The clinical disease and outcome are similar in goats. 1195
Sheep in the way to abort

1196
Mass aborted fetus

1197
 RVF cont’d---
In cattle the disease is somewhat less sever, with mortality rates in
calves and cows of 10 – 30%
However, 90 – 100% of pregnant cows abort.
Some animals develop emaciation with jaundice.
Necropsy findings
Extensive hepatic necrosis is the characteristic lesion in Rift Valley
Fever.
Some of additional necropsy findings are
Spleen enlargement
Hemorrhages in GIT and in subserosal
Non -specific lesions include
Congestion and petechiation in the heart, lymph nodes, gallbladder,
and alimentary tract.
Encephalitis, evidenced by neuronal necrosis and perivascular
inflammatory infiltration.
Diagnosis
As usual diagnose of RVF is made based on 1198
Hepatic syndrome, vasculitis and necrosis of the liver

1199
Intestine showing petechial haemorrhages on the serosal
surface

1200
1201
 RVF cont’d---
Epidemiology of the disease
Clinical signs
Necropsy findings
Laboratory diagnosis
In regions where this disease has not occurred RVF should be
suspect when there is an area outbreak of a abortion and neonatal
mortality in sheep and cattle coupled with an area outbreak of flu-
like disease in humans
Samples for confirmation of diagnosis are liver, spleen and brain.
Take liver, spleen and brain for histopathology and
immunohistochemistry.
The most common methods that are used for confirmatory diagnose
Virus isolation
Serology like direct FAT
Molecular techniques like PCR
NB. Note the zoonotic potential of this disease when handling
these specimens. 1202
 RVF cont’d---
RVFV is isolated in cell culture or mice.
RVFV is replicates in variety of cell culture such as
 Vero E6(African green monkey)
 BHK -21( Baby hamster kidney) cells
The virus causes CPE and plaques.
The isolated virus is identified by Virus Neutralization Test(VNT).
The past infection test can be known by using
 Ig M capture enzyme immunoassay on single acute sera.
 ELISA and haemagglutination inhibition on paired serum samples.
Differential diagnosis
RVF must be differentiated from blue tongue.
Treatment
Little attention has been given to the aspect of treatment of the
disease.
There is no any known treatment.
Prevention and control measures
Prevention and control measures of RVF comprises 1203
 RVF cont’d---
1. Live stock vaccination
2. Vector control
3.Quarantine
1. Livestock vaccination
There are two type of vaccine to control and prevent RVF.
These are
A. Live attenuated
B. Inactivated vaccine
A. Live attenuated vaccine
It is produced in mouse brain and in embryonated egg.
The vaccine is effective and inexpensive.
Live attenuated vaccine provide good protection which lasts for at
least 28 months.
But are not recommended for pregnant animals because they are
Abortigenic causing fetal death and
 Teratogenic anomalies
1204
 RVF cont’d---
B. Inactivated vaccine
It is killed-virus vaccines produced in cell culture.
It avoid the problem of abortion.
The immunity lasts for short period and require repeat
administration for good immunity .
So that annual vaccination of all dairy cattle and sheep is
recommended as a cost- effective control program in endemic
countries.
They are also recommended for pregnant and young animals.
What the problem not to be used widely because it is expensive.
2. Vector control
The use of mosquito larvicides and insecticides virtually impossible
in most areas of Africa.
This is because of
The involvement of wide range of vector species with different
habits and econiche preferences with wide geographical distribution.
The need to intervene through out long vector breeding seasons1205
 RVF cont’d---
But vector control would be a major element in control programs
were the virus to be introduced outside Africa.
3.Quarantine
Prevention of the introduction of Rift Valley Fever into countries
free of the disease requires
 The prohibition of the importation of all susceptible species from
Africa.
All necessary steps to prevent the introduction of infective insects
and infected biological materials should be taken.
However, the possibility of humans carrying the infection from
country to country is very rare but there is possibility.

1206
 Nairobi sheep disease is a vector born(tick-transmitted )disease
of small ruminants particularly sheep.
The disease is characterized by
Fever
 Haemorrhagic gastroenteritis
Prostration
Abortion of pregnant ewes with high mortality
Etiology
Nairobi sheep disease is caused by RNA virus called Nairobi sheep
disease virus (NSDV) which belongs to
Family Bunyaviridae
Genus Nairovirus
Nairovirus is Arbovirus meaning that the viruses are maintained in
arthropod – vertebrate – arthropod cycles.
It is possibly the most pathogenic virus known for sheep and
goat.
1207
Genetic and serologic data have shown that there are related viruses
 NSD Cont’d---
to Nairovirus in some parts of the world; like
 Dugbe virus that affect cattle in Nigeria
 Ganjam virus in Indian which affects sheep and goats
Epidemiology
It is endemic in East and Central Africa (Kenya, Uganda, Tanzania,
Ruanda, Somalia and Ethiopia).
Antibodies to the NSD virus and other bunyaviruses have been
detected in southern and northeastern Africa and Sri Lanka.
NSD affects sheep and goats.
The case-mortality rate is 30 - 90% in sheep but is lower in goats.
There are differences in susceptibility among different breeds of
sheep and goats.
Unlike in most other diseases, some indigenous breeds are more
susceptible than exotic breeds.
NSD virus does not affect cattle.
It can cause a mild febrile disease in humans.
Hence NSD is a zoonotic. 1208
 NSD Cont’d---
Source of infection
Sick animals
Adult ticks( long term carriage upto 2 years)
Means of transmission
It is tick transmitted(vector born) disease.

1209
Rhipicephalus appendiculatus

1210
Rhipicephalus appendiculatus on ears cow in Kenya

1211
Tropical bont ticks, Amblyomma
variegatum

1212
Adult tropical bont ticks, Amblyomma variegatum Fabricius,
feeding on the udder

1213
 NSD Cont’d---
The most common vector is the brown ear tick called
Rhipicephalus appendiculatus.
But other species like Amblyomma variegatum may be involved.
Transmission by Rhipicephalus appendiculatus can be both
Transovarian
Transstadially
Animals in endemic areas are usually immune .
The virus can persist in ticks for long periods, more than two years
in unfed adults.
The vertebrate reservoir host of the virus remain unknown.
The virus has not been found in wild ruminants or other animals in
endemic area.
Clinical disease occurs when susceptible animals are
Moved into endemic areas for marketing and grazing purposes
and/or
When there is a breakdown in tick control measures
For example in Kenya, sheep and goats acquire the infection when 1214
 NSD Cont’d---
they are transported from Northern districts to the Nairobi area.
Outbreaks occur outside endemic areas when
There is source of infection( infected animal and/or ticks)
 There is unusual increase in tick population brought about by
excessive or prolonged rains.
Clinical findings
After a short incubation period there is
A sudden onset of fever is followed by anorexia
 Nasal discharge
Dyspnea
Severe diarrhea because of hemorrhagic enteritis
Sometimes with dysentery
Abortion of pregnant ewes
 Lastly death within 3 - 9 days of 30 - 90 % affected animals.
Sometimes there is subclinical infections and recovered animals
become immune.
Ticks specially Rhipicephalus appendiculatus and other species1215
 NSD Cont’d---
like Amblyomma variegatum are likely to be found in the body,
especially in the ears and head of infected animals.
Necropsy findings
The necropsy picture is typical of a hemorrhagic diathesis consists
of:-
Hemorrhages on serous surfaces of visceral organs and on mucosal
surfaces, particularly the abomasum, colon and female genital tract.
Enlargement of mesenterial lymph nodes.
Hemorrhagic gastroenteritis becomes more obvious and there may
be zebra striping of the colon and rectum.
The uterus and fetal skin are hemorrhagic.
Common histopathological lesions outside the gastrointestinal tract
include : -
Myocardial degeneration
Nephritis and necrosis of the gall bladder
Diagnosis
As usual diagnosis of Nairobi sheep disease is made by complex1216
 NSD Cont’d---
method by considering
Epidemiology of the disease
Clinical signs of the disease
Gross pathologic examination
Laboratory
Specimens for laboratory diagnosis include
Uncoagulated blood taken during viremia
Mesenteric lymph nodes
Spleen
Confirmatory diagnose is made based on virus isolation and
identification of Nairovirus.
The virus is isolated
 In cell culture(Vero E6(African green monkey) or
 BHK -21( Baby hamster kidney) cells and/or
 In infant mice and the disease can be reproduced in susceptible
sheep.
Virus identification is done by immunological tests like: - 1217
 NSD Cont’d---
Immunofluorescence( direct)
Agar Gel Immunodiffusion(AGID)
CFT
 ELISA
The recommended serological test that is used to know the past
infection are
 Indirect fluorescent antibody test
Complement Fixation Test (CFT)
Indirect haemagglutination test
Differential diagnosis
Differential diagnoses include
Peste des petits ruminants (PPR)
Rinderpest
Rift Valley Fever
Heart water
 Parasitic gastroenteritis
Salmonellosis 1218
 NSD Cont’d---
All these diseases can be differentiated by laboratory diagnosis.
Treatment
There is no treatment for NSD
Prevention and control measures
The disease can be controlled by
1. Vector control and
2. Vaccination
1. Vector control
Vector control depends primarily on spraying and dipping of
animals with acaricides.
Controlling the main vector of the NSD called Rhipicephalus
appendiculatus is very important not only to control NSD.
But it also helps us to control economically very important
protozoan disease called East Coast Fever.
2. Vaccination
There are two types of vaccine to control NSD.
These are 1219
 NSD Cont’d---
Killed tissue culture vaccine
 Attenuated live virus vaccine
So that animals to be moved to endemic areas should receive either a
killed tissue culture vaccine or an attenuated live virus vaccine of
NSD.

1220
 Bluetongue is an arthropod borne viral disease of sheep
characterized by
Fever tat may lasts several days before hyperemia
Excessive salivation
Frothing at the mouth and nasal discharge
The tongue may become cyanosed, hence the name” blue tongue”.
Blue tongue virus affects cattle and goats and has subclinical form.
Etiology
Blue tongue is caused by RNA virus belongs to
 The family of Reoviridae
Genus Orbivirus
Within the BTV, there are at least 24 serotypes
There is considerable genetic variability within the serogroup.
This arises by genetic drift of individual gene segments as well as by
reassortment of gene segments when ruminant or the vectors are
infected with more than one strain.
1221
 Blue tongue Cont’d---
Epidemiology
Until the 1940 blue tongue was recognized only in Africa.
Then following a major epidemics in 1956 – 1957 in Portugal
and Spain the disease was recognized in USA, Middle East,
Asia and later in Australia.
Today blue tongue is recognized the disease is present in
most , if not all, countries in the tropics and subtropics.
Blue tongue is the most important disease in sheep.
Its severity depends on
 Serotype of the virus
Breed of the sheep

1222
 Blue tongue Cont’d---
Environmental stresses
Economic losses result from death and loss of conditioning in sheep
that recovered from the disease
But more importantly the disease has emerged as a major non –
traffic trade barrier.
That is suspicion of its presence is used at the convenience of
importing countries to limit live sheep, lamb and mutton imports.
In endemic areas, the infection is always present.
But clinical disease of the indigenous species is unusual.
Infection can occur only
When new BTV strains and
Non-indigenous susceptible species are introduced to the area
Epidemic zones also exist where infection and clinical disease occur
every few years.
Infection in these areas is highly focal and outbreaks occur when
Climatic conditions allow the vector to spread beyond its usual
boundaries and to infect susceptible ruminants. 1223
 Blue tongue Cont’d---
Incursive disease can occur in regions which do not normally
experience infection and may occur when
 The virus is introduced by windborne movement of infected
Culicoides with subsequent insect breeding in the end of winter
before ' die out' in the autumn and summer.
This method of spread is believed to have been the genesis of
several serious outbreaks of bluetongue in countries normally free of
the disease like
Outbreaks in Portugal in 1956, in Cyprus in 1977, in Turkey and
Greece in 1979-1980 and in Israel in 1960-1980.
Generally morbidity may be as high as 80% and mortality may reach
30%.
Source of infection
Infection occurs in a number of animals.
But significant disease occurs only in sheep.
Under natural conditions infection occurs in sheep and cattle and
rarely in goats. 1224
 Blue tongue Cont’d- --
Cattle are the major reservoir host for BTV for sheep.
But it is also recorded in elk, white-tailed deer, pronghorn antelope,
camels and other wild ruminants can be act as reservoir host.
Method of transmission
Bluetongue is not contagious disease.
It is transmitted by
1. Biologically by certain species of Culicoides
Virus in ingested blood infects cells of the mid gut by a receptor-
mediated process replicates and subsequently is released to the
salivary gland.
Once infected, they are infected for life - a period of several weeks.
The generation time in the summer and winter is averagely about 10
-14 days and more than 3 weeks respectively.
That means they can transmit the virus for this duration of life span.
Culicoides breed in damp, wet areas including streams, irrigation
channels, muddy areas and fecal runoff areas around farms, and
habitats for them exist on the majority of farm environments. 1225
 Blue tongue Cont’d---
Example species of Culicoides that are cattle associated, such as C.
brevitarsis, breed in cattle dung.
2. Other vectors may transmit the disease mechanically.
But are unlikely to be of major significance in disease epizootics.
These are
Argasid tick Ornithodoros coriaceus
The sheep ked (Melophagus ovinus)
The mechanical vectors ingests the virus when sucking the blood of
infected sheep and can transmit the infection in a mechanical
manner.
NB. Mosquitoes genus Aedes lineatopennis and Anopheles vagus
have been suspect in transmission of bluetongue virus
3. Venereal transmission
Bluetongue virus has been found in the semen of infected bulls
during the initial viremic period
So infection has been transmitted through bull semen to susceptible
1226
cows.
 Blue tongue Cont’d---
But it is unlikely that this is a significant mechanism of
transmission.
Risk factors of bluetongue
1. Pathogen risk factor
 2. Vector and environmental risk factors
3. Host risk factors
1. Pathogen risk factor
Genetic studies indicate that BTV tends to exist in discrete, stable
ecosystems.
BTV serotypes that circulate in one region of the world are largely
different from those in other regions.
 For example:- In Africa, serotypes 1, 16, 18, 19, and 24 are the
major serotypes that are circulating and causing bluetongue.
The bluetongue viruses are stable and resistant to decomposition
and to some standard verucidal agents, including sodium
carbonate.
They are sensitive to acid, inactivated below PH 6.0, and 1227
 Blue tongue Cont’d---
susceptible to 3% sodium hydroxide solution and organic iodides.
There are differences in virulence between serotypes and between
strains within serotypes.
2. Vector and environmental risk factors
The geographical occurrence of bluetongue serotypes varies and is
changing with time.
Different Culicoides species vary in susceptibility to infection.
Some known vectors are resistant to infection with some serotypes,
which in part explains regional differences in serotype occurrence.
Climate is a major risk factor as culicoides require warmth and
moisture for breeding and calm, warm, humid weather for feeding.
A cold winter or a dry summer can markedly reduce vector numbers
and risk for disease.
Moisture may be in the form of rivers and streams or irrigation that
helps the vector reproduction through out the year.

1228
 Blue tongue Cont’d---
But rainfall is the predominant influence and rainfall in the
preceding months is a major determinant of infection.
3. Host risk factors
Cattle are the reservoir and amplifying host and have a high titer
viremia.
Cattle appear to be much more attractive to Culicoides spp. and this
may enhance the importance of cattle as carriers.
Bos taurus breeds are more likely to be seropositive than Bos
indicus.
Bulls have a greater risk for infection than females or castrated
males.
Seroprevalence increases with age, probably a reflection of
increased duration of exposure.
All breeds of sheep are susceptible but to varying degrees.
Merinos and British breeds are more susceptible than native
African sheep.
There are also differences in age susceptibility to clinical disease
1229
 Blue tongue Cont’d---
which, inexplicably, vary with different outbreaks.
Exposure to solar radiation can increase the severity of the disease,
as can excessive droving, shearing, poor nutrition and other forms of
stress.
Pathogenesis
Bluetongue can be attributed to vascular endothelial damage
resulting in
Changes to capillary permeability and fragility
With subsequent disseminated intravascular coagulation and
necrosis of tissues supplied by damaged capillaries.
These changes result in
Edema
Congestion
Hemorrhage
Inflammation and necrosis.
Following infection into the skin, there is replication in the local
1230
 Blue tongue Cont’d---
draining lymph node and dissemination in mononuclear cells.
Circulating virus concentrations subsequently fall with the
appearance of circulating interferon and specific neutralizing
antibodies
With the viremia, there is localization of the virus in vascular
endothelium which causes endothelial cell degeneration and
necrosis with thrombosis and hemorrhage.
There is also the development of a hemorrhagic diathesis and
coagulation changes consistent with disseminated intravascular
coagulation.
Clinical signs
In sheep , bluetongue is characterized by
Fever that may last several days before hyperemia
Excessive salivation and frothing at the mouth with reddening of the
buccal and nasal mucosae
Swelling and edema of the lips, gums, dental pad and tongue occur
1231
and there may be involuntary movement of the lips.
 Blue tongue Cont’d---
 Excoriation of the buccal mucosa follows, the saliva becomes blood
stained and the mouth has an offensive odor.
Swelling and edema of the lips, gums, dental pad and tongue occur
and there may be involuntary movement of the lips.
 Excoriation of the buccal mucosa follows, the saliva becomes blood
stained and the mouth has an offensive odor.
Hyperemia and ulceration are also common at the commissures of
the lips, on the buccal papillae and around the anus and vulva.
Cyanosed tongue, hence the name” bluetongue”
There is facial swelling with extensive swelling and drooping of the
ears
Nasal discharge initially serous but become mucopurulent and
speckled with blood
Respiration is obstructed and stertorous and is increased in rate up to
100/min.
Marked loss of condition and sheep may die often through aspiration
pneumonia. 1232
 Blue tongue Cont’d---
 Some affected sheep show severe conjunctivitis, accompanied by
profuse lacrimation.
Foot lesions, including laminitis and coronitis, and manifested by
Lameness and recumbency, appear only in some animals, usually
when the mouth lesions begin to heal.
The appearance of a dark red to purple band in the skin just above
the coronet, due to coronitis, is an important diagnostic sign.
Wryneck, with twisting of the head and neck to one side, occurs in a
few cases, appearing suddenly around day 12.
This is apparently due to the direct action of the virus on muscle
tissue as is the pronounced muscle stiffness and weakness which is
severe enough to prevent eating.
Hyperemia of the skin may occur leading to “ wool break” some
week later.
Convalescence may be protracted and wool growth may be
impaired.
1233
As the result, partial or complete loss of the fleece is common and
 Blue tongue Cont’d---
causes great financial loss for the farmer.
Other signs during convalescence include
 Separation or cracking of the hooves and
 Wrinkling and cracking of the skin around the lips and muzzle.
Some bluetongue virus strains may cause
Abortion and
Congenital abnormalities like arthrogryposis, hydranencephaly and
ataxia.
In sheep in enzootic areas, the disease is much less severe and often
inapparent.
Two syndromes occur
An abortive form in which the febrile reaction is not followed
by local lesions and
Subacute type in which the local lesions are minimal, but
emaciation, weakness and extended convalescence are severe.
Infection in cattle and goats is usually subclinical.
But in Africa some cattle develop clinical signs similar to those seen
1234
 Blue tongue Cont’d---
in sheep.
The North American bluetongue viruses often cause a peracute fatal
hemorrhagic disease in cattle and goat.
Necropsy findings
The mucosal and skin lesions have already been described.
Other consistent lesions include
Generalized edema,
Hyperemia and hemorrhage and necrosis of skeletal and cardiac
muscles.
There is a most distinctive hemorrhage at the base of the pulmonary
artery.
 Animals with damage to esophageal or pharyngeal musculature
may have lung consolidation due to aspiration pneumonia
Aborted bovine fetuses should be examined for evidence of
hydranencephaly or porencephaly.
Diagnosis
Diagnose of Bluetongue as usual made by considering 1235
 Blue tongue Cont’d---
The epidemiology of the disease
 Clinical signs
 Necropsy findings
 Laboratory diagnosis
The main clinical pathology that are observed during acute
bluetongue infection are
 Increasement of Packed Cell Volume(PCV – value)
 Leukopenia due to lymphopenia followed by leukocytosis.
Confirmatory diagnose is made by isolation and identification of
bluetongue virus.
Specimens of choice for isolation are
 Blood taken in viremic stage
Chilled lung, spleen, CNS tissues and thoracic fluid from aborted
fetus
Specific diagnosis of bluetongue is made by
1.Isolation of the virus
2.Detection of viral antigen or nucleic acid 1236
 Blue tongue Cont’d---
3. Detection of specific antibodies in serum.
1.Isolation of the virus
Bluetongue viruses are often difficult to isolate in the laboratory.
The success of virus isolation is enhanced if blood is collected from
animals showing early clinical signs and if buffy coat is used as the
specimen of choice.
Bluetongue virus isolation commonly is carried out by
Cell culture(Usually by blind passage) or
Embryonated eggs
Less commonly, diagnosis of bluetongue is made by inoculation of
blood into susceptible sheep.
A positive test depends on
The appearance of diagnostic clinical signs and
Resistance to subsequent challenge with the bluetongue virus or
The significant increase in virus-neutralizing antibodies in the
recipient sheep.
1237
 Blue tongue Cont’d---
2.Detection of viral antigen or nucleic acid
The most common methods are the Immuno histochemical tests
including
Immunofluorescence
 Immunoperoxidase
 They use monoclonal antibody for rapid sensitive and specific
detection of antigen.
 In situ nucleic acid hybridization and PCR can be used for detection
of the virus
3. Detection of specific antibodies in serum.
A number of serological tests are available based on the detection
of group-reactive antibody or serotype-specific antibody.
Diagnosis of bluetongue by serology is imprecise, unless a rising
titer is demonstrated in acute and convalescent serum samples.
The commonly available serological tests to demonstrate
bluetongue virus antibodies by using monoclonal antibodies are
include 1238
 Blue tongue Cont’d---
CFT
AGID
ELISA
Among them Competitive ELISA has high sensitivity and
specificity.
Differential diagnose
Bluetongue in sheep and cattle must be differentiated from
Foot-and-mouth disease
Contagious ecthyma(orf)
Sheep pox
Treatment
Local irrigations with mild disinfectant solutions may afford some
relief.
Affected sheep should be housed and protected from weather,
particularly hot sun.
Fluid and electrolyte therapy and treatment to control secondary
infection may be desirable. 1239
 Blue tongue Cont’d---
Prevention and control measures
Prevention and control measures of blue tongue can be done by
1. Reduction of infection through vector control
2. Vaccination
3. Controlling international movement of livestock
1. Reduction of infection through vector control
Attempts to control bluetongue through
 Reduction of infection consist of reducing the risk of exposure to
infected Culicoides and
Reduction in Culicoides numbers
Neither of them are effective.
However, the risk of exposure is attempted to reduce by
Spraying cattle and sheep with repellents and insecticides and
Housing of sheep at night
During transmission periods avoidance of low, marshy areas or
moving sheep to higher altitudes may reduce risk
There is a high mortality in Culicoides that fed on cattle that have
1240
 Blue tongue Cont’d---
been treated with a standard antihelmintics dose of ivermectin.
Ivermectin has also a larvicidal effect in manure passed for the next
passed for the next 28 days for Culicoides that breed in dung.
2. Vaccination
Vaccination is the only satisfactory control procedure once the
disease has been introduced into an area.
Vaccination will not prevent or eliminate infection.
But it is successful in keeping losses to a very low level provided
immunity to all local strains of the virus is attained.
Current vaccines are usually polyvalent attenuated virus vaccines
and are in use in South Africa and Israel and available in other
countries.
The present Onderstepoort of South Africa Bluetongue vaccine
comprises three bottles (Vaccines A, B, and C) and includes the
following serotypes of BTV:
Bottle A: BTV serotypes 1, 4, 6, 12, and 14
 Bottle B: BTV serotypes 3, 8, 9, 10, and 11 1241
 Blue tongue Cont’d---
 Bottle C: BTV serotypes 2, 5, 7, 13, and 19
The three bluetongue vaccines are administered separately at
3-week intervals.
Reactions to vaccination are slight but ewes should not
be vaccinated within3 weeks of mating as an estrus often results.
Ewes should not be vaccinated within 3 weeks of mating as an
estrus often results.
Because the vaccine is live attenuated and can cause Early
Embryonic death or mortality(EED/EEM)
Annual revaccination 1 month before the expected occurrence of
the disease is recommended.
Immunity is present 10 days after vaccination.
So that early ring vaccination during an outbreak may
substantially reduce losses.
Lambs from immune mothers may be able to neutralize the
attenuated virus and fail to be immunized.
So that field strains may overcome their passive immunity. 1242
 Blue tongue Cont’d---
In enzootic areas, it may therefore be necessary to postpone
lambing until major danger from the disease is passed and lambs
should not be vaccinated until 2 weeks after weaning.
Rams should be vaccinated before mating time.
Live attenuated vaccines should not be used in pregnant ewes
Because of the risk of deformity in the lambs or embryonic death.
The danger period is between the 4th and 8th weeks of pregnancy.
The greatest incidence of deformities occurring when vaccination is
carried out in ewes pregnant for 5 - 6 weeks.
The preparation and use of attenuated vaccines against BTV is
problematic.
The neutralizing epitopes are highly conserved on some serotypes
But they are highly plastic on others.
Therefore it is necessary to monitor continuously the identity and
prevalence of the serotypes that need to be in the vaccine.
There are also concerns for the use of live vaccines to control insect-
borne diseases 1243
 Blue tongue Cont’d---
Because of the risk of the vaccine strain being transmitted, of being
exalted in virulence by passage, and of recombinants resulting in the
development of new virus strains with unwanted characteristics.
There is evidence for the emergence of a reassortment strain of BTV
Others require a negative test in conjunction with a period of
quarantine.
The introduction of bovine semen from low-risk areas is possible
 After negative test result of donors and
A prolonged storage period is accepted by most countries.
Most countries allow the importation of embryos.
in USA
However, living vaccines are used for practical reasons, including
the fact that inactivated
Vaccines of BTV do not provide protection against infection as it
produces low titer of antibody and not long lasting.
The difficulty in obtaining safe vaccines may be overcome by the
use of recombinant DNA technologies. 1244
 Blue tongue Cont’d---
3. Controlling international movement of livestock
Countries that are free of BTV infection have traditionally erected
barriers to avoid: -
animals from countries where the disease occurs.
Others have less severe restrictions and several procedures aimed at
permitting limited movement are in force; their stringency varies
with the importing country.
Some countries only require a negative serological test or series of
tests prior to movement.
Others require a negative test in conjunction with a period of
quarantine.
The introduction of bovine semen from low-risk areas is possible
 After negative test result of donors and
A prolonged storage period is accepted by most countries.
Most countries allow the importation of embryos.
1245
Here we focus on
5.1. Maedi - visna

1246
 Maedi – visna comprises two diseases of sheep those are caused by
one genus of virus.
The diseases are
Maedi
Visna
Maedi also called ovine progressive pneumonia.
Maedi are North American and European terms for slow virus
diseases of sheep in which a chronic progressive pneumonia is a
major manifestation.
As the name Maedi is derived from the Icelandic term for dyspnea.
While visna is associated with an Ovinelentvirus and is the
neurological manifestation of the disease.
Both the two disease forms may exist in one affected sheep and
called Maedi – visna.
Etiology
The disease is caused by RNA virus which belongs to
1247
 Maedi Cont’d---
Family Retroviridae
Genus Lentivirus
It consists of a variety of strains that vary in their ability to infect
target cells and cause disease.
There are neuroinvolvement and non - neuro involvement strains.
The non- neuro involvement strains cause maedi disease in sheep.
While the neuro involved strains cause visna disease.
Visna usually occurs in conjunction with maedi lesions in the lungs
a 10-18% of sheep with maedi have histological lesions of visna in
the brain.
Additional manifestations of infection are arthritis, indurative
mastitis, and ill-thrift.
There is a high degree of relatedness with its nucleotide homology
and serological properties with other lentivirus like
Caprine arthritis - encephalitis virus
Feline immunodeficiency virus
Equine infectious anemia virus 1248
 Maedi Cont’d---
Although they belong to a single virus species, isolates obtained
from naturally infected sheep are genetically heterogeneous,
antigenic drift is common.
The antigenic variation of the surface protein facilitates the
persistence of the virus in the host.
Epidemiology
Progressive pneumonic form of the disease called maedi was
reported early from South Africa and the United States.
But it now occurs in all major sheep producing countries with the
exception of Australia, New Zealand, Iceland and Finland.
Neurological form the disease called visna was originally recorded
in Iceland.
It was a significant cause of death of sheep in the country in 1950 -
1960s.
The two forms of diseases always occurs in association called
maedi- visna .
Despite the widespread occurrence ofmaedi-visna or ovine 1249
 Maedi Cont’d---
progressive pneumonia in many countries, visna is a ‘very
uncommon disease.
A significant occurrence of clinical disease of visna has seldom
been recorded in countries other than Iceland.
The reason for this is not known but might be due to
An increased susceptibility of the Icelandic breed of sheep to the
neurological form of the disease or
 The differences in maedi-visna virus strain neuroinvasiveness and
neurovirulence.
Maedi-visna was reported in many countries of the world with
largely been maedi occasionally with coexistent visna.
However, recently there have been the occurrence of visna with no
evidence of respiratory disease(maedi) suggesting that a neurotropic
strain might be emerging in different countries of the world.
Maedi – visna is a disease of sheep and rarely of goats.
Although rabbits are susceptible to maedi – visna, the infection
never cause disease state because. 1250
 Maedi Cont’d---
The disease is limited to the acute stage prior to the production
antibody.
Chronic infection does not occur as it does in sheep and goats.
That is why sheep and goats are the only species known to be
susceptible.
All breeds of sheep appear to be susceptible to infection.
But there may be differences in breed susceptibility.
Source of infection
The sources of infection are
 Clinically sick animals and/ or
 Chronic shedders
Transmission
The disease is spread by
Aerogenic (respiratory route)
 Ingestion of infected milk
Through different fomites
1251
Infection can also be transmitted intrauterine.
 Maedi Cont’d---
But the relative importance of this route in the spread of the disease
in flocks is debated.
Virus is also shed in the semen of infected rams that have
leukospermia.
Economic importance of the disease
The economic importance of maedi – visna comprise from
Lose associated with decreased life span
Lose due to mortality with clinical disease
Lose due to decreased value of cull animals and
Lose due to possible effects of subclinical infection on productivity
Clinical disease occurs in sheep 2 years old or older usually in
sheep 3 to 4 years.
The case fatality rate may reach100%.
Pathogenesis
The virus infects cells of the monocytes macrophage lineage and
attaches to cells by the binding of its envelope glycoprotein to
specific receptors on the cell surface. 1252
 Maedi Cont’d---
The virus replicates its RNA genome via a DNA intermediate
provirus which is integrated into the chromosomal DNA of
infected cells.
With initial infection there is virus replication; this is followed by
an immune response that restricts viral replication.
But fails to eliminate the virus completely.
Replication is restricted and does not proceed beyond the synthesis
of provirus in most infected cells.
There is persistence and replication of virus in the presence of
viral-specific immune responses, with the development of immune-
mediated lesions in various organ systems.
Persistent production of viral antigen results in lymphocytic
hyperplasia.
The infected macrophages in the various tissues are surrounded by
an inflammatory response creating a focus of mononuclear cell
aggregation.
The lungs, mammary gland, brain, joints, lymph nodes and blood 1253
 Maedi Cont’d---
vessels are affected by the maedi-visna/ ovine progressive
pneumonia viruses.
In the central nervous system there is infiltration of the meninges
and white matter with lymphocytes.
The demyelination that occurs in visna is believed to result from the
direct effect of the virus on oligodendrocytes and astrocytes
As well as being the result of an inflammatory response provoked by
the presence of viral antigen in these cells.
Similar infiltrations occur in the udder.
Lymphoid follicles are found in the alveolar parenchyma, often with
atrophy of the alveolar tissue.
Clinical signs
The incubation period in both maedi, visna and in maedi – visna
forms is long( more than a month)
Clinical disease, if it occurs, does not develop before 2 years of age.
That is why, most clinical sheep are older than 3 years.
In maedi forms, the clinical signs develop insidiously, progress 1254
 Maedi Cont’d---
slowly, and there is a long clinical course.
The earliest signs in maedi forms( progressive pneumonic form) are
Listlessness
Loss of body condition which progresses to emaciation which
results in increased cull rate of ewes because of poor condition.
 Signs of respiratory involvement are not evident in the initial stages
of the disease.
 But there is exercise intolerance and affected sheep will fall back
behind the flock when the flock is moved.
With the time the following clinical signs develop like
Dyspnea with an increase in respiratory rate (80-120 breathing
movement/min) at rest.
 Flaring of the nostrils or open mouth breathing develops later.
There may be coughing and some nasal discharge(but in most

1255
 Maedi Cont’d---
instances this occurs in sheep with secondary bacterial pneumonia).
There may be inflammation of the third eyelid.
Clinical illness lasts for 3-10 months and the disease is always fatal.
 Clinically affected sheep( pregnant ewes) are more prone to
diseases such as pregnancy toxemia.
In some sheep, clinical respiratory disease is minimal and the major
manifestation is wasting and the thin ewe syndrome.
In visna form, the disease has an insidious onset and the early
clinical signs include
Lagging behind the flock because of ataxia and body wasting.
The body wasting and the hind limb ataxia are progressive.
Affected animals show hypermetria and may stumble or fall as they
traverse uneven
ground or when making sudden turns.
There is no fever and a normal appetite and consciousness are
retained.
Additional signs include severe tremor of the facial muscles and1256
 Maedi Cont’d---
Knuckling of the distal limbs
So that the animal stands on the flexed tarsi.
Some animals may show a head tilt.
Aimless wandering and blindness occur in some sheep.
Necropsy findings
In maedi form of a disease, lesions may be present in the
Lungs and associated lymph nodes
Brain, joints, mammary gland and blood vessels
 But gross lesions in most sheep are confined to the lungs and, in
some cases in the mammary glands.
 In advanced cases, the lungs are larger and two to four times as
heavy as normal lungs.
They collapse much less than normal when the chest is opened and
are gray-blue to gray-yellow in color.
 There is a diffuse thickening of the entire bulk of both lungs and
the abnormal color and consistency are generalized and unvarying in
1257
all lobes.
 Maedi Cont’d---
Enlargement of the bronchial and mediastinal lymph nodes is
constant.
 There are frequently associated lesions of arthritis, encephalitis, and
mastitis.
Histopathological changes are characteristic of a chronic interstitial
pneumonia with proliferation of lymphoid tissue and the presence of
numerous lymphoid.
In visna forms of the disease
Muscle wasting and an interstitial pneumonia may be visible
But there are no gross changes in the central nervous system.
 The characteristic histological lesion is patchy, demyelinating
encephalomyelitis.
 Demyelination occurs in the white matter of the cerebrum and
cerebellum, and in the spinal cord.
 The inflammatory infiltrate is predominantly comprised of
lymphocytes and macrophages and pleocytosis is variable during
the course of the disease
1258
 Maedi Cont’d---
Diagnosis
Diagnose of maedi – visna is done based on
The epidemiology
Clinical signs
Necropsy finding
Laboratory
Confirmatory diagnosis is done by
Virus isolation and identification
Antigen detection by PCR
Samples for confirmatatory diagnosis( for virus isolation and
antigen detection) can be
Chilled brain
Cerebral spinal fluid(CSF)
Spinal cord
Synovial membrane
 Lung and mammary gland
Brain 1259
 Maedi Cont’d---
Specimens of choice for histopathology are
Formalin-fixed lung and bronchial lymph node
 Formalin fixed mammary gland and synovial membrane
Formalin fixed half of midsagittally-sectioned brain (LM) .
Flock status with respect to the presence or absence of infection and
the determination of the infection status of an individual sheep
currently relies on serological testing.
The most common serological tests are
Agar gel immunodiffusion (AGID) test
 ELISA
NB. The specificity and sensitivity of most currently available
serological tests are inadequate to determine the infection status of
an individual and the results of flock tests of a potential source of
replacement sheep should be used coupled with an examination of
postmortem records in the potential source flock if available.
Differential diagnose
1260
Maedi form of a disease must be differentiated from many chronic
 Maedi Cont’d---
diseases of sheep and goats that cause pneumonia and chronic
wasting conditions.
Among them the most common are
Parasitic pneumonia
Chronic suppurative pneumonia
Post-dipping pneumonia
Enzootic pneumonia
 Melioidosis
 Johne's disease
Caseous lymphadenitis
Visna is a sporadic disease of mature sheep with an insidious onset
of muscle wasting, progressive ataxia and having a long clinical
course.
These characteristics differentiate it from other diseases of sheep
manifest with ataxia.
So that visna must be differentiated from
1261
Scrapie
 Maedi Cont’d---
ewes at birth.
However giving them no colostrum from infected mother but bovine
colostrum
And rearing them on milk replacer quite separately from other
of particular value where there is a requirement to retain genetic
lines in the eradication procedure.
But it has its own drawn back because
It is very labor-intensive and expensive.
This can create a considerable potential for reinfection of the
artificially reared flock.
2. Test and cull
This involves the detection and culling of seropositive animals.
This method is the preferred method where lateral transmission is
the dominant mode of transmission in the flock.
All sheep (and goats) on the farm are serologically tested annually
or twice a year,.
Then seropositive animals and their progeny of less than 1 year of
1262
 Maedi Cont’d---
sheep are culled.
This method is effective in establishing an infection-free sero
positive animals.
Testing is continued semi - annually or annually until there are at
least two consecutive negative tests.
The offspring of older seronegative ewes are kept for
replacements.
Future flock introductions with both methods should be from a
seronegative flock.
Other control procedures
Control procedures that attempt to limit or delay the spread of
infection, and consequently the occurrence of clinical disease within
an infected flock, have a limited success.
In flocks that have a high incidence of clinical disease, the
determination of the age of onset of clinical disease and the
establishment of a culling policy based on this information can
reduce the economic impact of the disease.
Flock introductions should be from seronegative flocks where 1263
 Maedi Cont’d---
In the absence of such programs, severe restriction of inter-farm
movement and outdoor housing may limit the spread of the disease.
The fact that some sheep in infected flocks retain freedom from
infection in the face of continual exposure to infection suggests that
there is a genetic resistance to infection.
The identification of these determinants may prove to be the future
method for the control of this disease.
Implementation of strong biosecurity policy
Once infection is introduced to a flock it is difficult and expensive
to eradicate.
So that all efforts should be directed to prevent its introduction.
This can be achieved by having strong biosecurity in the farm.
Rams and replacement ewes should be acquired from accredited
free flocks in countries where the disease is registered.

1264
Here we give attention to
6.1. Classical swine fever
6.2. African swine fever

1265
 It is also called Hog cholera.
It is highly contagious and highly fatal disease of swine caused by
pestivirus .
The disease is characterized in peracute and in acute form by
 Viremia
Haemorrhagic diathesis
Abortion
Death
In subacute and chronic form by
Pneumonia
Colitis
Etiology
Hog cholera is caused by RNA virus belongs to
 Family Flaviviridae
 Genus Pestivirus
There is only one antigenic type with many numbers of strains
1266
which have variable virulence.
 Hog Cont’d---
The pestivirus has the following strains
Strains of high virulence
Strains of mild virulence
Strains of low virulence
The virus has an antigenic relationship to the bovine virus diarrhea
virus (BVDV).
Epidemiology
The pig is the only domestic animal species naturally infected by
the virus.
All breeds and ages are susceptible
However adults are more likely to survive an acute infection than
young.
The disease originated in the United States but is now almost world
wide in distribution.
The disease is currently endemic in most countries of South
America and the Far East except Japan and Korea.
The disease has also concentrated in certain parts of Europe where
1267
 Hog Cont’d---
the pig populations are intense and live in close proximity to wild
boar .
Thus there may be transmission and persistence of the virus within
the wild boar population.
As pig production continues to become intensified the disease has
been more difficult to control.
When a virulent strain of the virus infects a susceptible population,
the disease usually occurs in epidemics.
Virulent strain has often with a morbidity of 100% and a case-
fatality rate approaching 100%.
However, in recent years outbreaks of a relatively slowly spreading.
Because the mild form of the pestivirus has caused great concern in
many countries.
Mild strain of pestivirus produces a mild viremia with widespread
antigen distribution.
But without clinical signs, except lesions of hemorrhagic
dermatitis. 1268
 Hog Cont’d---
It produces an antibody response and trans- placental infection.
The disease associated with strains of low virulence may be
unnoticed in growing in adult pigs.
But the infection can be associated with perinatal mortality,
abortions and mummification of fetuses.
Source infection can be
Sick animals
Recovered animals which are carriers
Products from the above animals
Infection is usually acquired by
Ingestion
Inhalation
Direct animal to animal contact
Infected pigs shed a large amount of the virus in all normal
secretions - nasal, salivary, urinary, fecal
They are important in transmission when there are in clinical signs
of the disease. 1269
 Hog Cont’d---
The virus is excreted in the urine for some days before clinical
illness(incubation period) appears and for 2 - 3 weeks after clinical
recovery.
The resistance and high infectivity of the virus make spread of the
disease by
Inert materials(fomites)
Uncooked meat
Artificial insemination (This is because the virus probably infects
spermatogonia).
 Contaminated instruments and drugs
Because the common practice of not changing syringes and needles
between farm visits constitutes a major risk when viremic animals
are present.
The most common cause of dissemination occurs through the
movement and sale of infected or carrier pigs in communal market
places.
1270
 Hog Cont’d---
NB. In areas free of the disease, introduction is usually by the
importation of infected pigs or the feeding of garbage containing
uncooked pork scraps.
Risk factors for occurrence of CSF
The most common risk factors are
Animal risk factor
Pathogen risk factor
It is known that CSF
 More affects finishing pigs than sows and boars.
 Susceptible pregnant sows if exposed to less virulent strains of the
virus may remain clinically healthy but infection of the fetuses in
uterus is common.
So that the virus may be introduced into susceptible herds by way
of these infected off springs.
The genotype of pigs may influence the outcome of hog cholera
virus infection.
1271
 Hog Cont’d---
Pure bred pigs resulted in acute fatal infections
While cross-bred pigs experienced acute, chronic and latent
infections.
Virulence characteristics the virus is the pathogen risk factors of
the virus.
The most virulent strains produce clinical disease in pigs of all ages.
While the less virulent strains cause only mild clinical disease or
disease restricted primarily to fetal and newborn piglets.
It is probable that this variance has always occurred in field strains
of the virus .
But the use of inadequately attenuated live virus vaccines is also a
contributory factor.
The occurrence of variation in virulence and antigenicity has been
recognized as a cause of failure of vaccination and vaccine
breakdowns.
The other pathogen risk factor is the resistance of the virus to
different temperature, chemical and PH of the environment. 1272
 Hog Cont’d---
The virus can be killed by
Boiling
5% cresol or 3% sodium hydroxide
Sunlight through 45 – 60 minutes
But it persists in meat which is preserved by salting, smoking and
particularly by freezing.
The virus can survive in infected uncooked ham pork for at least 84
days.
It survives with PH ranges from 3 to 11.
Persistence in frozen meat has been observed after 4.5 years. The
virus persists for 3- 4 days in decomposing organs
The virus persists for 15 days in decomposing blood and bone
marrow.
Economic importance
Classical swine fever(Hog cholera ) has been responsible for large
economic losses in the swine industry worldwide.
It is considered to be the most important disease of pigs in1273the
 Hog Cont’d---
European Union.
The economic loss because of Hog cholera comprises
Losses due to the death of pigs
Direct costs of transport and destruction of infected herds.
 Compensation loss for farmers
Loss for controlling and preventation of the disease(loss for
disinfection of premises and for vaccination program).
Loss due to reduced growth rate of affected pigs.
Loss for medicaments since susceptibility of recovered or partially
recovered pigs to other diseases have increased
Because most of recovered or partially recovered pigs are very
susceptible to secondary infection such as enzootic pneumonia of
pigs.
Pathogenesis
CSF virus reproduces mostly in lymphoid reticular organs.
After exposure, the tonsil is the primary site of virus invasion
following oral exposure. 1274
 Hog Cont’d---
Primary multiplication of the virus occurs in the tonsils, beginning
within several hours after infection.
After 6 hours the virus through lymphatic system enters and can be
found in regional lymph nodes.
Then after 24 hours you may found the virus in the blood .
Eventhough the CSF virus is panthrophic virus , it more replicates
in organs and tissues those are rich in lymphoid – reticular cells like
Lymph nodes
Tonsils
Spleen
Liver
 Peyer's of Patches
Bone marrow liver
Then the virus exerts its pathogenetic effect on endothelial cells,
lymphoreticular cells, macrophages and epithelial cells.
This effect on the vascular system results in the characteristic
lesions of congestion, hemorrhage and infarction from changes in 1275
 Hog Cont’d---
blood vessels.
Thrombosis of small and medium-sized arteries is another feature.
Vascular changes are most severe in the lymph nodes, spleen,
kidneys and gastrointestinal tract.
Pathogenic effect of the virus on endothelial cells cause occlusion
of blood vessels.
This results the destruction of the wall of blood vessels and mass
haemorrhagiae in different organs.
Because of the disturbance of blood circulation of lymph nodes,
intestine and other organs, there may found necrosis on mucous
membrane of GIT and infracts in spleen in other parenchymatous
organs
The virus affects blood forming organs as the result the animal
develops anemia and leucopenia.
Atrophic process in lymphoid – macrophage tissue ends with
blockage of immune system.
Lymphocyte apoptosis in which activation of induced 1276
 Hog Cont’d---
programmed cell death is one of the key features of CSF infections.
All this results in exposure of affects pigs to secondary bacterial
infection that cause pneumonia, colitis (pasteurellosis and
salmonellosis).
The virus can infect the brain and spinal cord forms perivascular
infiltrate and cause aseptic encephalomyelitis.
That is why there may be depression, mania and tonic convulsion
as clinical feature in CSF.
In acute form of a disease, death of animals is due to the
disturbance of the function of blood forming and blood circulatory
organs .
In subacute and chronic form of the disease death of animals is
mostly related to the disturbance of metabolism which ends with
blockage of the immune system and failure of the natural resistance
of pigs.
That is why pigs are susceptible to secondary conditional
pathogenic bacterial infections like salmonellosis and pasteurellosis.
1277
 Hog Cont’d---
Clinical findings
Incubation period of CSF is 5 – 8 days.
Rarely, it can be short upto 3 days or long upto 2 – 3 weeks.
Depending on the length of incubation period CSF has the
following forms
Peracute
Acute
Subacute
Chronic
In apparent forms
Depending on the types of signs predominate during infection, CSF
can be conditionally classified into
Septicemic manifestations
Nervous manifestations
Breathing manifestations
Intestinal manifestation
Atypical form of the disease 1278
 Hog Cont’d---
In peracute form of CSF, the main clinical findings are
Haemorrhagic spots on skin
Vomiting
Death after 1- 2 days
In acute form of CSF the common clinical signs are
 Pyrexia constant type that can reach upto 40 – 41 0C
Depression
 Standing of pigs in a drooped position with their tails hanging
They are disinclined to move and, when forced, do so with a
swaying movement of the hindquarters
Vomiting
Anorexia
Constipation followed by diarrhea
Later a diffuse purplish discoloration of the abdominal skin
Conjunctivitis is usual and in some pigs the eyelids are stuck
together by dried, purulent exudate.
Nervous signs often occur in the early stages of illness and include
1279
 Hog Cont’d---
Peracute form of CSF occurs very rarely in young pigs.
At the beginning of an outbreak, young pigs may die per acutely
without evidence of clinical signs.
But if the affected pigs survive they may develop
Fever (410C and above)
Tachycardia and polypnea
Abortion
Rhinitis and bleeding of the nasal cavity.
On skin surface of femoral, abdominal cavity, neck and on the base
of ear develops pustules contain yellowish exudate.
After some times petechial haemorrhagiae may develop which can
connect them selves to form large black spot on it.
This black spot on the skin never temporarily removed from the
skin when you press by figures and it is used as method of
differential diagnosis from swine erysipelas .
NB. Infection with Salmonella choleraesuis may also be
potentiated by hog cholera infection and the two diseases in 1280
 Hog Cont’d---
combination can result in high mortality.
This progressive weakness results in difficulty in breathing and
heart function which results in
Cyanosis of the skin of nostrils, ears, ventral abdomen and distal
parts of legs.
Leucopenia
In peracute and acute form of CSF, some animals may have
nervous manifestation with clinical signs like
 Lateral recumbency
Tetanic convulsion for 10 - 15 seconds followed by a clonic
convulsion of 30 - 40 seconds.
Apparent blindness, stumbling and allotriophagia have also been
observe
Subacute form of CSF has breathing or intestinal manifestation.
Because the virus affects either the lung and thoracic cavity or
intestine.
When the virus affects the breathing organ, it has the following 1281
 Hog Cont’d---
clinical manifestations
Intermittent fever
Anorexia
Pneumonia
Difficulty in breathing
Pain around thoracic cavity
When the virus affects the intestine, the following are the main
clinical features
Intermittent fever
Anorexia
 Enterocolitis
Constipation followed by diarrhea
Most of the animals may die and those which recover become
carrier animals.
In chronic form of CSF is caused by low virulence strains of virus.
The incubation period is longer may reach 2 months and above.
The main clinical signs in chronic CSF are 1282
 Hog Cont’d---
Persistent mild fever
Depression
Periodic diarrhoea
Anorexia
 Unthriftiness
Coughing
 Characteristic skin lesions including alopecia, dermatitis, blotching
of the ears and a terminal, deep purple coloration of the abdominal
skin.
CSF may cause reproductive failure without other clinical evidence
of disease within the herd.
It may occur in pregnant sows
 When inadequately protected pregnant sows are exposed to
virulent virus and/or
When susceptible pregnant sows are vaccinated with live
attenuated vaccines or
When they are exposed to low virulent field strains. 1283
 Hog Cont’d---
Infection of the pregnant sows may result in with no clinical signs
other than a mild pyrexia.
But it may be followed by a
 High incidence of abortion
Low litter size
Mummification
Stillbirth and anomalies of piglets
Persistent congenital infection of piglets is characterized by
Persistent viremia
Continuous virus excretion
Late onset of disease, with death occurring 2 - 11 months after birth
Atypical form of CSF occurs when piglets and young pigs soon
after weaning which have maternal antibodies are infested with
high virulent strains or
When the piglets and young pigs are infested with low virulent
strains of virus.
The disease is characterized by subacute and chronic forms with1284
 Hog Cont’d---
typical clinical signs of
Anorexia
Conjunctivitis
Subcutaneous haemorrhagiae
Most of the animals recover slowly.
However, a few of the affected animals may develop pneumonia
and gastroenteritis and they may die
This is because of the complication with secondary bacterial
infection.
Necropsy findings
In peracute cases there may be no gross changes at necropsy.
In the more common acute form, there are many submucosal and
subserosal hemorrhages.
But these are not the constant necropsy findings in CSF.
The hemorrhage results from erythrodiapedesis and increased
vascular permeability, probably aided by mast cell degranulation.
1285
The hemorrhages are most noticeable under
 Hog Cont’d---
The capsule of the kidney
 The ileocaecal valve
 The cortical sinuses of the lymph nodes
 the bladder and larynx.
The hemorrhages are usually petechial and rarely ecchymotic.
The lymph nodes are enlarged
Spleen contain marginal infarcts.
Infarction in the mucosa of the gallbladder is a common necropsy
finding in CSF
However, it is not constant finding.
But appears to be an almost pathognomonic lesion for CSF.
There is congestion of the liver and bone marrow and often of the
lungs.
Circular, raised button ulcers in the colonic mucosa are usual
necropsy findings in CSF.
But it is difficult to distinguish from those of salmonellosis.
The above gross necropsy findings are fairly typical in the case1286
of
 Hog Cont’d---
hog cholera.
However, they cannot be considered as diagnostic unless
accompanied by the clinical and epizootological evidence of the
disease.
Because they can occur in other diseases, particularly
salmonellosis caused by Salmonella cholera suis.
There are characteristic microscopic lesions of a non-suppurative
encephalitis during nervous form of the disease.
Histologically, the main site of tissue injury is the
reticuloendothelial system.
There is always a progressive lymphoid depletion and mucosal
necrosis.
The lymphoid depletion is probably caused by viral induced
apoptosis but not by direct apoptosis.
In the chronic form of the disease, ulceration of the mucosa of the
large intestine is usual.
Secondary pneumonia and enteritis commonly accompany the 1287
 Hog Cont’d---
primary lesions of hog cholera.
Diagnosis
Diagnose of CSF can be done based on
Epidemiology of the disease
Clinical signs
Necropsy findings
Laboratory diagnose
A valuable antemortem diagnostic test is the total and differential
leukocyte count.
In the early stages of the disease there is a marked leukopenia.
There is specifically a granulocytopenia caused by a bone marrow
Atrophy.
In the late stages of hog cholera, a leukocytosis due to secondary
bacterial invasion may develop.
B-lymphocytes, T-helper cells and cytotoxic T-cells are the most
affected by the virus.
The loss of the circulating B lymphocytes was consistent with the
1288
 Hog Cont’d---
failure to generate a circulating neutralizing antibody.
Virulent strains produce a more reduction in B-lymphocytes than
do mild forms.
This can be of value in differentiation from bacterial septicemias
But it should not be used as the sole method of differentiation.
The disease is confirmed by isolation and identification of the
virus.
Samples for confirmation of diagnosis can be
Heparinized blood( taken at viremic stage)
 Lymph nodes
Tonsil
Spleen
Distal ileum
Skin
Tongue
Brain
If there is doubt about the presence of the virus, you can confirm
1289
 Hog Cont’d---
by injecting filtrates from tissue suspension in non – immunized
piglets.
If CSF virus is present the pig lets will die after 7 – 10 days.
Confirmatory diagnose is done by isolation and identification of the
virus .
The virus can be isolated by using piglets.
A positive diagnosis of hog cholera is difficult to make without
laboratory confirmation.
The isolated virus can be detected by using different serological and
molecular techniques including
Fluorescent antibody techniques
Antigen-capture ELISA
Agar gel precipitation test
Molecular techniques like RT –PCR
A comparison of diagnostic tests shows that the best results are
detected by RT-PCR
There are many pest viruses like BVDV and ASFV that must be 1290
 Hog Cont’d---
differentiated from CSF
Differentiation of swine fever virus from other pestiviruses can be
done using
Reverse transcriptase polymerase chain reaction ( RT - PCR) assay
Different serological tests those are used to detect antibody
including
Fluorescent antibody test
Virus neutralization test
Tissue culture serum neutralization test
Indirect ELISA
Differential diagnosis
A positive diagnosis of hog cholera is difficult to make without
laboratory confirmation.
The major diseases which resemble hog cholera · include:
 Salmonellosis
 Erysipelas
Pasteurellosis 1291
 Hog Cont’d---
Encephalomyelitis caused by other viral diseases and
salmonellosis
African swine fever
Salmonellosis can be differentiated from CSF because
It has less severity than CSF
It usually accompanied by enteritis and dyspnea
Erysipelas is differ from CSF because
Skin haemorrages have characteristic diamond shaped
The subserous hemorrhages are likely to be ecchymotic rather than
petechial.
 The ecchymotic skin haemorrhagiae temporary disappears when
you press by your figure .
However, in CSF the haemorrhagiae are more petechial and do not
disappear temporarily when you press by your fingure.
Pasteurellosis can be differentiate from CSF because
 In pasteurellosis respiratory signs are predominate
 Lesions in pasteurellosis are predominately pleuropneumonia at1292
 Hog Cont’d---
necropsy
In addition, epidemiological considerations, hematological and
bacteriological examination will usually help us to differentiate the
above diseases from CSF.
Viral encephalomyelitis and salmonellosis cause similar nervous
signs and can be differentiated in laboratory.
African swine fever (ASF) differs from CSF by its greater
severity.
Apart from this it is almost impossible to differentiate from hog
cholera without laboratory testing.
Treatment
There is no effective treatment for CSF after the clinical signs are
developed
So that clinically sick pigs must be killed as soon as possible to
control the wide spread of the disease.
However, it is possible to use hyperimmune serum.
Hyperimmune serum may be of value in the very early stages of1293
 Hog Cont’d---
the illness if given in doses of 50 -150ml subcutaneously.
But it has more general use in the protection of in-contact animals for
preventation measure than for treatment of sick pigs.
Control and preventation measures
The methods those are used in the control CSF include
Eradication
Control by vaccination
In areas where there are effective barriers to control the reintroduction
of CSF, control of the disease can be established: -
By eradication of the disease by slaughter methods
This method of controlling CSF economically feasible and usually
desirable in such areas.
In contrast, in areas where there is no barriers because the pig industry
forced them to have movement of pigs within-country and across-border.
It may not be practical or economically feasible to institute a slaughter
eradication program.
So that in area where there is continuous outbreak of CSF, control even
eradication of the disease possible only by vaccination. 1294
 Hog Cont’d---
Control of CSF varies depends on the epidemiology of the disease
in the area.
So that the control strategy of CSF includes
1. Control of outbreaks in hog cholera free areas
2. Control where hog cholera is endemic
1.Control of outbreaks in hog cholera free areas
In areas where the disease does not normally occur, eradication is
possible by
Slaughter of all in contact and infected pigs is recommended.
The pigs are slaughtered and disposed of preferably by burning.
All herds in the area should be quarantined and no movement of
pigs permitted unless for immediate slaughter.
All vehicles used for the transport of pigs, all pens and premises
and utensils must be disinfected with strong chemical disinfectant
such as 5% cresyic acid.
Contaminated clothing should be boiled.
Entry to and departure from infected premises must be carefully1295
 Hog Cont’d---
controlled to avoid spread of the disease on footwear, clothes and
automobile tires.
Legislation prohibiting the feeding of garbage or commanding the
boiling of all garbage before feeding must be enforced.
2. Control where hog cholera is endemic
Control of CSF in hog cholera endemic area must be directed to
Control CSF in wild life reservoir like in the wild boar population.
Select the best vaccine and identification of carrier animals
 Notifying the outbreaks on time
Stop purchase and sale of infected or in-contact pigs
Stop feeding of uncooked garbage
Oral vaccination of wild boar usually reduces the presence of CSF
But only a low rate of wild boar (30 - 35%) become seropositive.
Because it was shown that more than 50 % of the wild boar did not
feed on the vaccination baits and therefore did not become immune.
In endemic areas, control is mostly a problem of selecting the best
vaccine. 1296
 Hog Cont’d---
The first group is the classical live group containing attenuated
virus which are preferred
Because live attenuated virulent virus vaccines produce a solid
immunity within just a few days(3 - 5 days)
They give life long protection.
But they are capable of introducing the infection and of actually
causing the disease when vaccination 'breaks' occur.
The reaction to live virus vaccine may be severe and the
susceptibility of pigs to other diseases may be increased.
Eradication of the disease is impossible while the use of this type of
vaccine is permitted.
Second group of live vaccines aimed as marker vaccines which is
prepared based on the E2 protein.
There appears to be no complete protection against congenital
infection.
The immunity lasts only for about 1 year.
Failure to notify outbreaks; to stop purchase and sale of infected1297or
 Hog Cont’d---
in-contact pigs, to stop feeding of uncooked garbage can be
improve
Through education by showing to the farmers the highly
contagious nature of the disease and the ease with which it can be
spread by the feeding of uncooked garbage and the purchase and sale
of infected or in-contact pigs.
The common practice of sending pigs to market as soon as illness
appears in CSF endemic area is one of the major methods by which
hog cholera is spread.
When an outbreak occurs in a herd, the immediate need is to
prevent infection from spreading further.
This can be best achieved by
 Removing the source of infection and
 Increasing the resistance of in-contact animals
Removing of the source of infection can be achieved by
 Isolation of infected animals.
1298
 Hog Cont’d---
Applying suitable hygienic precautions to prevent the spread of
infections on boots, clothing and utensils
 Disposal of carcasses by burning
Disinfection of pens.
The pens should be scraped, hosed and sprayed with 5% cresylic
acid solution or another suitable disinfectant like queenicides that
contains quaternary ammonium.
The resistance of in-contact animals can be achieved by
administration of
Hyperimmune serum
 Available vaccines
So that pigs in the affected pen should receive serum (20-75 ml
depending on size).
While pigs in unaffected pens should be vaccinated.
There is also inactivated vaccine which is used in control CSF.
It has been the one of the most widely used vaccine to eradicate the
disease in many countries of Europe. 1299
 Hog Cont’d---
It is completely safe vaccine
But its has its own demerits, because
The immunogenicity is poor.
Immunity does not develop until 12 days after vaccination.
Its duration is short and booster injections are required for
maintenance

1300
 African swine fever has another names like
African pig disease
Wart hog disease
African swine fever is highly contagious viral disease of pigs in
Africa characterized by
Fever
Cyanosis of skin
General haemorrhagiae in internal organs
High case fatality up to 90 – 100%
ASF is an OIE list A disease.
It is indistinguishable in the field from classical swine fever as both
cause hemorrhagic diatheses with high fatality.
However, ASF is associated with a totally different virus.
Etiology
It is associated with a DNA virus which is the sole member of
the family Asfarviridae.
1301
 ASF Cont’d---
The virus is large icosahedral cytoplasmic DNA virus.
It is the only known DNA arbovirus.
Morphologically, it is similar to the iridoviruses
But resembles the pox viruses in genome construction and gene
expression.
There are different strains of virus.
They are
Virus strains those causing highly lethal form of ASF
Virus strains causing sub-clinical form of ASF
Tissue culture adapted strains ASFV
Epidemiology of ASF
African swine fever is indigenous to the African continent where it
affects mostly wild pigs.
The wild pigs are
Warthogs
Bush pigs
Forest hogs 1302
 ASF Cont’d---
These wild pigs act as reservoirs of the virus.
The ASFV can cycles between the pigs and the ticks.
So that ASFV is arbovirus.
It was always considered to be a disease of Sub –Saharan Africa.
Because until 1957, African Swine Fever (ASF) had not occurred
outside the African continent.
But in last 60 years several outbreaks were registered in many
countries of the world including Spain, Portugal, Italy, France,
Cuba, Brazil and many countries of Latin America and Australia.
Twenty years ago, there was an outbreak in Belgium(March 1985).
The source of infection was thought to be pork imported from
Spain which was fed to only one boar.
The source of infection are
 Sick pigs
 Carrier animals
 Reservoirs(wild pigs)
The disease can be transmitted to susceptible animals 1303
 ASF Cont’d---
 Vectors
 Ingestion
 Aerosol
The main vectors to transmit ASF in Africa is a soft tick(argasid)
tick Ornithodoros moubata.
The viremic wart hog is a source of infection for the ticks.
The virus can be maintained in wart hog- associated argasid ticks by
Transstadial transmission mechanism
Transovarial transmission mechanism
Sexual (male to female but usually not vice versa) transmission
mechanism.
It needs to replicate in the mid-gut epithelium of the tick for
successful ASF infection of the tick.
The tick has a wide distribution in Africa south of the Sahara.
The main habitat is in burrows which are inhabited by the wart hog.
In some areas where infected wart hogs are common but where
Ornithodoros moubata is apparently absent there are other soft ticks
1304
 ASF Cont’d---
for transmission of ASFV.
These are
Ornithodoros. savignyi
Ornithodoros porcinus
ASFV is also found the bush pig (Potamochoerus porcus)
It can transmit the virus to ticks following infection which may be viremic
for 35 - 91 days after infection.
In Africa, the virus is maintained primarily by a cycle of infection between
wart hogs and soft ticks (Ornithodoros moubata).
The European vector of the virus are the soft tick Ornithodoros erraticus.
It can maintain and transmit the virus for at least 300 days.
Many species of soft ticks in genus Ornithodoros that are feed on
pigs may be capable of acting as vectors of the virus in many countries of
the world.
However, there is a possible existence of potential vectors among the
other bloodsucking arthropods and should not be ignored.
Many researchers think that ASFV can be transmitted by hog louse
Haematopinus suis.
1305
 ASF Cont’d---
After infection the virus excreted together with blood, feces, urine ,
secretion of the mucous membrane and saliva.
In addition to vectors, susceptible animals are infested by
Indirect contact by infected pens
Ingestion of contaminated feed and water
Feeding uncooked garbage containing infected pig material
Aerosol through respiratory system
Through contaminated wounds created by fighting
Ris factors
Pathogen risk factors
Animal risk factors(immune mechanisms)
Recent studies of ASFV have suggested that the virulence may
depend on their ability to regulate the expression of macrophage
derived cytokines.
Macrophage derived cytokines in turn regulate Th1 and Th2
responses and control the host protective responses.
The less virulent cultures of ASFV with macrophages produce more1306
 ASF Cont’d---
TNF -α , IL-6, IL-12, and IL-15.
However, virulent strains
Inhibit the production cytokines
Affects chemotactic responses
Phagolytic capacity
Reduction in the release of toxic oxygen radicals.
The virus is highly resistant to
Putrefaction
Heat (it will survive 2 h at 56°C)
Dryness
 Survives in chilled carcasses for upto 6 months and at 4°C for 2 years.
Antibodies against the African swine fever virus occur in the
colostrum of sows previously infected with the virus.
The antibodies can be transferred passively to nursing pigs.
Experimentally passively transferred virus specific immunoglobulins
alone will protect swine against lethal infection with a highly
virulent homologous strain of the virus.
1307
 ASF Cont’d---
Pathogenesis
The virus invades pigs mainly through the tonsils and respiratory
tract.
ASFV replicates first in the lymphoid tissues of the nasopharynx.
Then the virus through lymphatic system reach to all organs of the
organism and cause generalized viremia.
The virus is panthrophic virus and can replicate in different organs.
However, the virus mostly affects lymphoid organs and endothelium
of blood vessels.
In the blood the virus affects mostly the mononuclear phagocytic
cells like monocytes, other reticular cells and macrophages causes
necrosis and lysis of these cells.
Then after the virus replicates in lymph nodes and endothelium of
blood vessels.
Pathogenetic factors that play in development of the disease are
mass destruction of cells by ASFV.
Because of the replication ASFV in the cells. 1308
 ASF Cont’d---
As the result there is high production of biological active substances
like
Pyrogenic substances
Serotonin
Histamines
Lymphokines
These biological active substances cause disturbance in
thermoregulation and cellular metabolism and cause mass cellular
death.
The virus causes hemorrhages through its effect on haemostatic
mechanisms by affecting vascular endothelium.
After about 4-5 days the vascular damage extends to the basement
membranes and death ensues usually because of the serious edema
and hemorrhage.
The mechanisms related to hemorrhage consist of
Activation and extensive destruction of monocytes and
macrophages. 1309
 ASF Cont’d---
Disseminated intravascular coagulation
Infection and necrosis of megakaryocytic cells
Early in the infection there is prolongation of coagulation times due
to
Inhibition of fibrin formation
Thrombocytopenia
Finally because destruction of endothelium of blood vessels,
disseminated intravascular coagulation lead the development of:-
Hemorrhages
Serous exudates
Infarction
 Local edema
Engorgement of tissues
Since the virus destructs immune cells by inducing apoptosis of the
immune cells.
So that the organism becomes immune incompetent and can be
exposed to other secondary bacterial complication. 1310
 ASF Cont’d---
All clinical forms of the disease are characterized by extensive
hemorrhages at necropsy.
It is this feature which often establishes a presumptive diagnosis in
the field.
The lymphopenia which is so characteristic of the disease is due to a
significant increase in lymphocyte death by virus induced apoptosis.
The virus can cross the placenta and replicate in fetal tissues and
cause abortion.
However the pregnancy failure is probably the result of the effects
of the virus infection on the dam more than from direct viral
damage to the placenta or fetus.
Clinical findings
By external clinical signs of the disease, it is difficult to differentiate
ASF from CSF.
Length of incubation period, form and severity of ASF depends on
 Virulence of the virus
 Dose of the virus 1311
 ASF Cont’d---
Routes of infection
The incubation period of ASF after contact exposure varies from 5 -
15 days.
Depending on the severity of the disease, ASF can have
Peracute form
Acute form
Subacute form
Chronic form
Latent form
Peracute form of ASF occurs very rarely and have the following
clinical signs:-
Fever(40.5 – 420C)
Dysponea
The animals may die after 1 – 3 days because of shock .
The acute form of ASF is the most common form in pigs
It has an average incubation period of 5 – 15 days.
The animals die in an acute state of shock characterized by a 1312
 ASF Cont’d---
disseminated intravascular coagulation with multiple hemorrhages
in all organs.
However, if the affected pigs survive, they show the following
clinical signs: -
Fever which reach upto 40.5 0C
Tachycardia
Cyanosis of the skin
Depression, anorexia and huddling together
Disinclination to move, weakness and incoordination
Extreme of the hindquarters with difficulty in walking
Coordination remains in the front legs
Serous to mucopurulent nasal and ocular discharges
Dyspnea and cough
Diarrhea sometimes dysentery
Vomiting and pregnant sows usually abort
Purple discoloration of the skin may be present on the limbs, snout,
abdomen and ears. 1313
 ASF Cont’d---
NB. Abortion because of ASF may occur in all stages of gestation
about 5 - 8 days after the infection commences or after 1-2 days of
fever.
The clinical signs of ASF in subacute form of the disease is similar
to that of acute form.
But subacute form of ASF has less degree of manifestations.
The clinical signs in subacute form ASF can be seen for twenty days
following infection.
The body temperature of affected animals can be 40.5 – 420C for
one week following infection then reduces to 40.5 0C
Most of the pigs die while the rest of pigs can be chronically sick.
Chronically affected cases are/ have
Febrile intermittently
Low growth rate
 Emaciated
Signs of broncho pneumonia
 Edematous swellings over limb joints called polyarthritis 1314
 ASF Cont’d---
Edematous swelling under the mandible
Necrosis of ears, skin of lower limbs, shoulders and head of
animals.
In latent form of ASF, pigs are carrier of the virus.
However, due to different stress factors the virus replicate and
excretes/ secrets from the pigs and become source of infection.
Recovered pigs have no lesions suggestive of the disease
But they may be viremic for several weeks.
These persistently infected pigs would pass routine antemortem
inspection at slaughter and potentially infectious offal and carcass
trimming could be fed unknowingly to other pigs.
Necropsy findings
Gross changes at necropsy resemble closely those found in hog
cholera.
However, in the acute ASF, the lesions are more severe than in
CSF.
The most common gross findings in acute form of ASF are 1315
 ASF Cont’d---
The presence of black colour with cyanotic spots on the skin of ear,
ventral abdominal cavity and on the internal lateral surfaces
femoral region.
Distended blood vessels.
Rarely hematoma around inguinal canals.
Haemorrages and hematoma in different muscles
Swollen and hemorrhagic in gastro hepatic, mesenterial and renal
lymph nodes and they may resemble spleen.
Other gross lesion in acute form of ASF are
Sub capsular petechiation of the kidneys(the renal hemorrhages are
considered almost to be pathognomonic for ASF).
Ecchymoses of the cardiac surfaces
Ecchymoses on various serosae
 Pulmonary edema with hydrothorax
Splenomegaly of spleen is usual( but in contrast to hog cholera,
splenic infarcts are rarely seen)
1316
 ASF Cont’d---
The gallbladder is edematous and hemorrhagic
There is severe submucosal congestion in the colon as CSF
However, button ulcers in the large intestine in ASF are less
common when compare with CSF.
An encephalitis may be present with lymphoid infiltration during
ASF( but is generally less severe than that of hog cholera).
Histologically the lesions are more diagnostic.
So that the most common histopathological changes are
 Destruction of the mononuclear phagocyte system and then infects
megacaryocyte, tonsillar crypt cells, renal cells, hepatocytes and
endothelial cells
Destruction of monocytes/macrophages is visible in the lymph
nodes, the spleen and the bone marrow.
Destruction of hepatocytes in the liver
In subacute and chronic ASF the gross and histological lesions
have less severity and seems that of CSF.
1317
 ASF Cont’d---
Diagnose
Diagnose of ASF can be done by considering the
Epidemiology of the disease
Clinical signs of the disease
Necropsy findings of the disease
Laboratory findings
In epidemiology of the disease consider
 The trade and economical relation ship of the country for those
countries which are free from ASF.
The severity of epidemics
The fatality of the disease
Although most of the clinical signs are similar to CSF, you have to
consider the presence of
 Pyrexia constant type for 3 – 6 days.
 Hemodynamic disturbance
 Cyanosis of ear and skin of abdomen
Edema of lung 1318
 ASF Cont’d---
Diarrhea rarely dysentery ( bloody)
Bloody nasal and oral discharges
From pathomorphological changes consider
Splenomegaly( 1.5 – 2 times)
Haemorrhagiae in kidneys with petechiation in nephron.
 Haemorrhagic pneumonia with edema in connective tissues found
between doles of the lung.
Haemorrhagy in portal, mesenterial and renal lymph nodes
 Accumulation of serous – haemorrhagic exudate in thoracic,
abdominal cavities and in pericardium of heart.
In laboratory findings, consider the
Reduction of total leukocyte count by 40 - 50% in the fourth day
of fever.
 Presence of lymphopenia
 Increasement immature neutrophils
 Elongation of the clotting times of blood at about 5 days post-
infection.
Thrombocytopenia that is detectable from day 6 after infection.1319
 ASF Cont’d---
However the diagnose is confirmed by isolation and identification
of the virus from appropriate specimens.
The appropriate specimens are
Heparinized blood taken at viremic stage
Spleen
 kidney
Submadibular and abdominal lymph nodes
Tonsil
Visceral fluid samples
For histopathology parallel collect formalin-fixed
 Spleen
Lung
Lymph nodes
Kidney
Liver
Colon
Cecum 1320
 ASF Cont’d---
Brain
It is possible to use these specimens for
Histopathology
 To detect the virus by Immuno Histo- Chemistry( IHC)
Antigen can be detected from specimens by direct FAT in tonsil
and madibular lymph node within 24 - 48 hours of infection.
The indirect FAT and direct FAT are commonly carried out on
pooled visceral fluid samples.
ASFV can be isolated in culture cells of
 Leucocytes from swine
Swine bone marrow
The isolated virus can be identified by serological and molecular
techniques including
FAT (Fluorescence Antibody Test)
CFT(Complement Fixation TEST)
RDP(Reaction diffuse precipitation)
While PCR is the common molecular technique used to identify 1321
the
 ASF Cont’d---
isolated virus
Nowadays ASFV reduces its virulence and most of the infections
have either subacute or chronic or in latent forms.
So that the presence of the infection in the herd can be diagnosed by
collecting appropriate specimens from necessarily slaughtered pigs
by appropriate serological tests like
Reaction haemadsorption
Direct and indirect FAT
CFT
RDP
The past infection of our herds can be diagnosed by detecting
specific antibody.
As antibody to ASFV may be detected within 7 days of infection.
This antibody can be detected by serological tests like
 ELISA (highly sensitive and specific)
RIA(Radio Immuno Assay)
Indirect FAT 1322
 ASF Cont’d---
CFT
RDP
NB. If you have a doubt in isolation and identification of the virus,
you can inoculate your specimens to pigs those are vaccinated
against CSF.
Differential diagnose
ASF must be differentiated from
Hog cholera(CSF)
Aujeszky's disease(Pseudo rabies)
Pasteurellosis
Erysipelas
ASF always confuse with hog cholera.
So that very careful examination is required to differentiate the
two.
The two diseases can be differentiated by their
Severity
Histopathological change under microscope 1323
 ASF Cont’d---
Haemadsorption in cell culture of leucocytes
FAT
Test in vaccinated laboratory animals.
Clinically, the illness from ASF is much shorter than in hog
cholera.
Gross necropsy changes are similar but more severe than those of
hog cholera.
More recently, reliance has been placed on the demonstration of
hemadsorbing activity with virus from suspected outbreaks grown
on pig leucocyte tissue cultures.
ASF can be easily differentiated from Aujesky's disease(Pseudo
rabies) in that
Aujesky’s disease affect not only pigs but many domestic and wild
animals(cattle, dogs, cat and rats).
 Aujesky’s disease more has paralytic form with out haemorrhagic
diathesis.
Erysipelas is differ from ASF because 1324
 ASF Cont’d---
Skin haemorrages have characteristic diamond shaped
The subserous hemorrhages are likely to be ecchymotic rather than
petechial.
The ecchymotic skin haemorrhagiae temporary disappears when
you press by your figure .
Pasteurellosis can be differentiate from ASF because
 In pasteurellosis respiratory signs are predominate
 Lesions in pasteurellosis are predominately pleuropneumonia at
necropsy
Treatment
There is no treatment for ASF.
Treatment is forbidden by law in most countries of developed
world.
Control and preventation measures
Slaughter affected pigs and killing of ticks as quickly as possible.
However, control and eradication of ASF is difficult because of
the:- 1325
 ASF Cont’d---
Lack of an effective vaccine
Transmission of the virus in fresh meat and cured pork products
Recognition of persistent infection in some pigs, particularly wild
feral pigs, possibly warthogs and bush pigs
Clinical similarity of hog cholera and African swine fever
Recognition that in some parts of the world soft ticks of the genus
Ornithodoros (erraticus, moubata, porcinus) are involved in the
biological transmission of the disease and can remain carriers for
long periods (possibly 5 years).
Prevention of introduction of the disease to free countries is crucial
to control ASF.
It can be done based on the prohibition of importation of live pigs
or pig products from countries where African swine fever occurs.
However, if an outbreak occurs control must consist of prevention
of spread by
Quarantine
1326
 ASF Cont’d---
Slaughter of infected and in -contact animals and
Suitable hygienic precautions including using of appropriate
disinfectants and acaricides.
The disease is virtually uncontrollable when swine farms are not
intensive.
Because in semi – intensive and extensive swine farms, pigs have an
access
to communal grazing.
The virus is highly resistant to external influences including
chemical agents.
That is why common disinfectants never kill the virus.
So that the most practical disinfectant to use against the virus is a
strong solution of caustic soda.
The main difficulties encountered in the eradication program of ASF
in many countries of the world.
The highly resistant nature of the virus that makes it to be infective
1327
in contaminated sties for periods exceeding 3 months.
 ASF Cont’d---
The persistence nature of the virus in recovered pigs
Indiscriminate use of attenuated vaccines, which fostered the
development of carrier pigs
The current eradication program of ASF consists of the following: -
 Depopulation of herds with clinical disease
 Serological surveillance of all sows and boars in every herd
Improvement of sanitary conditions of housing
 Improved hygiene (safe disposal of manure, vehicle disinfection,
insect and rodent extermination)
 Veterinary control of all swine livestock transfers (with individual
identification of every animal moved for finishing or breeding
purposes)
 Health certification of every animal used for herd replacement
 Destruction of every seropositive animal
 Formation of mobile veterinary field teams exclusively dedicated
to support the program
1328
 ASF Cont’d---
There are several vaccines that have been used in ASF preventation
and eradication programs.
The vaccines are
Inactivated virus vaccine and
Modified live virus vaccines
Inactivated vaccine produce short duration of protection.
So that it needs booster doses .
The modified live virus vaccines provide some protection better
than inactivated ones.
But the results following their use have been neither satisfactory
nor safe.
So that they have the two disadvantages, includes
Confounding laboratory tests
Producing' carrier' pigs.
1329
1330
 4.1. Introduction to prions
4.2. Common diseases of large animals caused by
prions
 4.2.1. Bovine Spongiform Encephalopathy(BSE)
 4.2.2. Scrapie

1331
4.1. Introduction to prions
Prions are normal cellular proteins that have under gone
conformational change.
As the result of post translational processing of a normal cellular
protein they become pathogenic.
The normal protein, called PrPC term for the normal cellular
isoform of the prion protein.
An abnormal folded isoform, designated PrPSC of a host encoded
cell-surface glycoprotein (prion protein, PrPC) accumulates during
disease and is associated closely with infectivity.
The function of PrPC is not known and the mechanism by which
PrPC is converted to PrPSC is uncertain.
The normal prion protein is composed of about 208 amino acids.
It is encoded in the genome of most mammals and expressed in
many tissues, specially in
Neuron
Lymphoreticular cells
1332
The abnormal isoform of the protein called scrapie isoform of the
 Prions Cont’d---
prion proteins and are designated by PrPSC .
The amino acid sequence of PrPC and the abnormal isoform of the
protein called PrPSC term for the scrapie isoform of the prion
protein in a given host is identical.
Only the confirmation of PrPSC is changed from a structure made
up predominately of α – helices( predominate in prion proteins
PrPC) to one made up predominately of β – sheets(predominate in
abnormal isoform proteins called PrPSC).
That is why PrPSC which are rich in β - sheets are isolated as
insoluble aggregates during different transmissible spongiform
encephalopathies (TSEs) diseases of animals and human beings.
The agent that cause transmissible spongiform encephalopathies
(TSEs) is the abnormal isoform of the prion protein and that, in the
infected host, this can recruit further alternatively folded prion
protein.
With this theory the long incubation period of prion diseases
reflects the rise in level and deposition of PrPSC in a variety of 1333
 Prions Cont’d---
tissues, including brain, eventually resulting in fatal spongiform
encephalopathy.
Scrapie in sheep has been recognized for over 200 years.
But the recent epidemics of Bovine Spongiform
Encephalopathy(BSE) has focused public attention and scientific
research on the TSEs.
Many research works with scrapie, and other TSEs showed that
transmission was successful by crude or purified extracts of brain or
other tissues from affected animals and human beings.
The infective agent is very resistant to ionizing and ultraviolet
irradiation and to reagents that damage or modify nucleic acids.
This, along with other experimental findings, has led to proposals
that the infectious agent in scrapie, and other TSEs, is the PrPSC
itself.
But not a small, unconventional virus or virions as previously
proposed.
The structure of the infecting PrPSC is believed to imprint upon the
1334
 Prions Cont’d---
normal cellular precursor PrPC, resulting in a change to the
abnormal isoform which is protease-resistant and accumulates in
cells.
Recently, a monoclonal antibody was developed that can
discriminate between the normal and disease specific form of PrP.
The monoclonal antibody specifically precipitates bovine, murine
and human PrPSC but not PrPC.
The results of attempts at inter-species transmission of these
diseases are variable.
Frequently they do not transmit.
But there are a few case when the TSEs agents can be transmitted
to other species .
Successful primary transmission between different mammalian
species typically requires larger dose to effect disease than would be
required for transmission to the same species.
Parenteral or intracerebral routes are required and success is greater
with young animal recipients. 1335
 Prions Cont’d---
This is the so-called 'species barrier.
When using transmission studies to detect the presence of one of
these agents optimal sensitivity is with a recipient host the same
species is necessary.
To solve the problem of species barrier, it is possible to use
transgenic mice.
That is why the 'gold-standard' technique for the diagnosis of TSE
agents is the passage of tissue in panels of inbred mice.
4.2. Some common diseases large animals caused by prions
The transmissible spongiform encephalopathies (TSEs) are a group
of progressive neurological disorders that are transmissible and
affect a number of animal species and humans.
They are non-febrile with long incubation periods and a long
course of disease.
Some of the diseases of animals are caused by prions are given in
table below.

1336
 Prion cont’d---
4.2. Some common diseases large animals caused by prions
Diseases Affected hosts Sources of infection
Bovine Spongiform Cattle Acquired
Encephalopathy(BSE) Bovine spongiform encephalopathy
prion contamination of meat - and -
bone meal ; some vertical transmission
from cow to calf
Scrapie Sheep, goats Natural
and Moufflon Not certain, possibly scrapie prion
contained in feed, but more likely by
direct contact and contamination of
pastures by placentas and fetal tissues
Transmissible mink Mink Acquired
encephalopathy Scrapie prion contamination of sheep
(TME) carcasses and offal fed to mink
Feline spongiform Zoo cats (puma, Acquired
encephalopathy(FSE) cheetah and From prion contamination of meat
domestic cats)
1337
4.2.1. Bovine Spongiform Encephalopathy(BSE)
Bovine spongiform encephalopathy (BSE) is an afebrile
neurological disorder affecting adult cattle.
It causes subacute transmissible spongiform encephalopathy.
It occurs as the result of the exposure of cattle to animal protein
feeds containing the scrapie agent.
The disease is of considerable importance mainly because
 It has zoonotic potential
It has also a potential to spread into many countries
The cost of controlling the disease is very high
Etiology
BSE is a prion-associated transmissible spongiform encephalopathy.
What are the prions that cause BSE?
There are many debate as to prions that cause BSE because
 1.There is evidence of characteristics that distinguish it from
conventional scrapie strains.

1338
 BSE Cont’d---
2. A strain of scrapie that has modified to infect cattle or
3. Whether it is originated from a sporadic spongiform
encephalopathy pre-existent in the cattle population.
In view of the high susceptibility of certain African ungulates and
zoo carnivores to infection, it has also been postulated that the
agent could have entered into meat-and-bone meal from the
carcass of an animal that died in a zoo or a safari park in the UK.
A further hypothesis promotes meat and bone meal from the
Indian sub-continent as a source.
Although there are many debates for the cause of BSE, there has
been no conclusion and the source may never be known.
Epidemiology
In Great Britain, the first known clinical case probably occurred in
1985.
But the BSE Inquiries considered it likely that there had been
several undetected cycles of BSE in the South West of England in
the 1970s and early 1980s. 1339
 BSE Cont’d---
The disease was subsequently recognized in Switzerland, Portugal
and France in the early 1990s and is now widespread in the world.\
Cases have occurred in imported British cattle in Oman and the
Falkland and Channel Islands Countries that have had cases of BSE
in native cattle are
Austria, Belgium, Canada, Czech Republic, Denmark, Finland,
Germany, Greece, Ireland, Israel, Italy, Japan, Luxembourg,
Netherlands, Poland, Portugal, Slovakia, Slovenia, Spain,
Switzerland, United Kingdom, and the United States of America.
The OIE Terrestrial Animal Health Code describes five BSE risk
categories for countries.
In order of increasing incidence of BSE these categories are
 BSE free countries
BSE provisionally free countries
Minimal BSE risk countries
 Moderate BSE risk countries
 High BSE risk countries 1340
 BSE Cont’d---
These classifications can be important in international live animal;
animals’ products and bi products trade.
There has been no apparent breed predisposition.
In both herd types, the risk for cases increased significantly with
increasing herd size.
Epidemiological studies suggest that most affected cattle have been
infected at their calves stage.
Risk of BSE is greatest in the first 6 months of life and between 6
and 24 months of age.
Risk is related to feeding patterns of proprietary concentrates.
Adult cattle are at low risk for infection.
Other species spongiform encephalopathy have been identified in
seven species of ungulates in zoos or wildlife parks in Great Britain
since the occurrence of the disease in cattle.
These animals had been fed meat and- bone meal.
But the apparently shorter incubation period suggests that they
might be more susceptible to infection than cattle. 1341
 BSE Cont’d---
There is evidence for horizontal transmission.
Feline spongiform encephalopathy (FSE) is also recorded in
domestic cats in Great Britain since 1990 and in zoo felids.
The zoo felids had been fed cattle carcasses unfit for human
consumption.
The zoo had a history of BSE in exotic ruminants and fed culled
carcasses to other zoo animals.
In mice with the agents associated with these encephalopathies in
zoo ungulates and felids suggest that they are the same strain as
causes BSE.
Source of infection
The source of infection in both countries is believed to have been
meat-and-bone meal imported from Great Britain.
Means of transmission
Ingestion of proprietary meat-and-bone meal.
Meat-and-bone meal is manufactured by the rendering industry
from tissues discarded in slaughterhouses, and also from down and1342
 BSE Cont’d---
dead livestock.
The outbreak of BSE in Britain was temporarily preceded by a
change in the method of processing of meat-and-bone meal to a
continuous process with a cessation of the use of hydrocarbon fat
solvents.
It is postulated that this change, coupled with the high sheep to cattle
ratio in Great Britain, allowed the greater survival of a scrapie-like
agent in scrapie-infected sheep carcasses processed to meat-and-
bone meal to reach a level that was infective for cattle.
The marked fall in disease incidence following the introduction of
the feed ban in 1987.
UK substantiated the importance meat-and-bone meal as the major
method of infection.
Calves born after the ban in the UK and in other counties a number
Have developed the disease.
Most of these were born in the years immediately following the ban
1343
and their numbers have decreased in subsequent years.
 BSE Cont’d---
livestock.
The outbreak of BSE in Britain was temporarily preceded by a
change in the method of processing of meat-and-bone meal to a
continuous process with a cessation of the use of hydrocarbon fat
solvents.
It is postulated that this change, coupled with the high sheep to
cattle ratio in Great Britain, allowed the greater survival of a
scrapie-like agent in scrapie-infected sheep carcasses processed to
meat-and-bone meal to reach a level that was infective for cattle.
The marked fall in disease incidence following the introduction of
the feed ban in 1987.
UK substantiated the importance meat-and-bone meal as the major
method of infection.
Calves born after the ban in the UK and in other counties a number
Have developed the disease.
Most of these were born in the years immediately following the ban
and their numbers have decreased in subsequent years. 1344
 BSE Cont’d---
This may be the result of exposure, at birth, to high infectivity in
birth products as there is no evidence for Infection and transmission
in embryo transplants.
Risk for occurrence of BSE
There are many risk factors for occurrence of BSE.
Among them the most common are
Changes in the method of processing meat and-
bone meal
Importing of latently infected cattle
Importing of infected meat-and-bone meal.
These risks can be substantially avoided by
Prohibiting the feeding meat-and-bone meal to cattle.
Prohibition of importation of latently infected cattle and meat and
bone meal.
NB. Countries with largely pastoral cattle are at low risk.
1345
 BSE Cont’d---
Zoonotic implication
BSE is a zoonotic disease because it causes CJD in human beings.
CJD (Creutzfeldt–Jakob disease): - It is a degenerative
neurological disorder that is incurable and invariably fatal.
CJD is at times called a human form of mad cow disease (bovine
spongiform encephalopathy or BSE).
Pathogenesis
Pathogenesis of BSE is not well studied.
Clinical findings
BSE is insidious in onset and the clinical course progresses over
several weeks, varying from 1 to 6 months in duration.
There is a constellation of clinical signs with alterations in
behavior, temperament, posture, sensorium and movement.
The predominant neurological signs are
Having apprehensive behavior

1346
 BSE Cont’d---
Hyperesthesia
Unusual kicking
Head tossing when haltered
Change in posture and movement and being ataxic
Behavioral changes are gradual in onset and include changes, such
as reluctance to pass
 Affected cattle are disoriented and may stare, presumably at
imaginary objects, for long ‘ periods.
 There is hyperesthesia to sound and touch, with twitching of the
ears or more general muscle fasciculation and tremors.
Other changes in temperament include
 Avoidance of other cows in loose housing
 Having antagonistic behavior to herd mates and humans when in
confined situations.
Affected animals may kick during milking and show resistance to
handling.
Relatively early in the course of the disease there is 1347
 BSE Cont’d---
Hind limb ataxia with a shortened stride
Swaying gait
Difficulty in negotiating turns
In the later stages of the disease the cow may show sign of
Knuckling
 Stumbling and falling with subsequent difficulty in rising
Progressive weakness with ataxia and weight loss.
Necropsy finding
There are no abnormalities in gross pathology and diagnosis is
dependent upon histological findings.
Major changes are in the brain stem and the pathognomonic lesion
is a bilaterally symmetric intracytoplasmic vacuolation of neurons
and gray matter neuropil.
The occurrence of vacuolation in the solitary tract and the spinal
tract of the trigeminal nerve in the medulla oblongata is the basis of
statutory diagnosis of the disease
Histopathological findings are diagnostic in many cases. 1348
 BSE Cont’d---
But can be supplemented with the immunodetection of PrPSC in
brain tissue by in situ immunohistochemistry and Western immuno
blots.
Diagnosis
No confirmatory ante-mortem test available to diagnose BSE.
But there are clinical methods to diagnose BSE in live animals
even if they are not confirmatory.
Clinically BSE in live animals can be diagnosed by the reaction of
animals to
Sudden noise
Sudden light
Sudden movement
Sudden touch
The reaction of BSE affected animals to sudden noise is tested by
clanging two metal objects together out of sight of the animal.
The test is called the bang test.
The reaction of BSE affected animals to sudden light is tested by
1349
 BSE Cont’d---
But can be supplemented with the immunodetection of PrPSC in
brain.
The reaction of BSE affected animals to sudden movement is tested
by waving a clipboard towards the cow from a short distance .
The test is called clipboard test .
The reaction of BSE affected animals to sudden touch is tested by
touching the animal on the hind limbs by soft stick.
The test is called stick test.
Abnormal reactions to the above tests include
Being startled
Head tossing
Salivation
Snorting
Running away or panicky circling
 Kicking out on touch
There are a number of validated rapid tests for BSE in dead animals.
Recently it has been established that the sensitivities of detection
1350of
 BSE Cont’d---
BSE-specific PrP by
 Immunohistochemistry.
Detection of infectivity by mouse inoculation
The specimens of choice for confirmatory test for histopathology is
formalin-fixed brain, including midbrain and entire medulla
oblongata.
The above specimen can be used for histochemistry.
The above specimens must be collected and submitted to laboratory
for mice inoculation.
NB. Note the zoonotic potential of this disease when handling
carcass and submitting specimens.
Differential diagnosis
BSE should be considered in the differential diagnosis of any
progressive neurological disease in cattle.
Primary diseases that must be differentiated at BSE based on
clinical signs include
1351
 BSE Cont’d---
Hypomagnesaemia
 Nervous acetonemia
 Rabies
Lead poisoning
Listeriosis
Polioencephalomalacia
Tremorogenic toxins.
Bovine amyloidotic spongiform encephalopathy
Treatment
There is no treatment for the disease.
Prevention and control measures
The prevention and control measure of BSE must focus on
1. Detection of BSE in surveillance and control programs
2. Measures to protect human health
1.Detection of BSE in surveillance and control programs
Detection of BSE can be done by using
 Passive surveillance 1352
 BSE Cont’d---
 Active surveillance
1. Passive surveillance has been used in many countries.
Suspect disease is notifiable with compulsory slaughter and
compensation and disposal of the carcass by incineration.
1. Active surveillance is directed at a targeted proportion of culled
animals.
In active surveillance animal manifesting neurological disease,
rabies-suspects but negative for rabies, and a proportion of cattle,
or all cattle, over 24 to 30 months (depending on country) that are
presented for slaughter for human consumption.
In slaughter cattle, the sampling frame is set to detect BSE at a
prevalence rate of one mature animal in a million mature animals.
The ability to conduct active surveillance, particularly on
slaughter cattle, has been allowed by the development of rapid
tests that can be conducted and read while the carcass is being
held so that positive test cattle are not released for human
consumption. 1353
 BSE Cont’d---
Positive rapid tests need to be conformed by histology and
immunohistochemistry.
Control of BSE in cattle
Control of BSE has the following assumptions
Infection and disease in cattle is introduced through feeding
contaminated feed containing infected meat-and-bone meal or
greaves
 The source of infection to cattle can be eliminated by effective
prohibition on feeding infected feed
There is no significant horizontal or vertical transmission.
Based on this, most countries have established a ban on the feeding
of ruminant protein to ruminants.
There is however a strong argument for banning all mammalian
protein for feeding to all livestock.
The experience of several countries with animals that were 'born
after the ban shows that cross contamination in feed mills can occur.
1354
Removal of Specified-Risk-Materials (SRM) like
 BSE Cont’d---
Brain
Spinal cord
Eyes
Tonsil
Thymus
Spleen
 Intestines
All these SRM must be removed from cattle carcasses.
This reduces the risk of BSE agent being in the subsequent
rendered carcass.
Measures to protect human health
Measures to protect human health from BSE are
High risk animals, such as downer cows should be kept out of the
humans food chain and not rendered for meat-and bone meal.
Infection is present in the tissues listed as SRM (brain, spinal cord,
eyes, tonsil, thymus, spleen, and intestines) must be removed from
the carcass at slaughter 1355
 BSE Cont’d---
The removal of SRM also protects against the risk posed by cattle
that may be incubating the disease yet do not show any symptoms.
Together with a ban on products such as mechanically recovered
meat that could be contaminated with SRM, excluding SRM from
the human food chain is the most important food safety measure to
However, this may not be sufficient and additional cares must be
taken like
Stopping of method of slaughter with captive bolt guns that can
spread brain within the carcass by blood into the pulmonary tissues
and elsewhere.
 Stopping of method of splitting the carcass and spinal cord that can
result in significant carcass contamination and contamination of the
slaughterhouse environment.
 Based on ' transmission and infectivity experiments cattle under 30
months of age are considered to have very low risk of being infected
in BSE non- endemic countries.
But it can be a risk in endemic countries with cattle over 1356
 BSE Cont’d---
this age.
Some countries with a high incidence of BSE have banned cattle
over 30 months for human consumption.
4.2.2. Scrapie
Scrapie is a non-febrile, fatal, chronic disease of adult sheep and
goats, characterized clinically by pruritus and abnormalities of
gait and has very long incubation period.
It is the prototypic disease for a group of diseases known as
Transmissible Spongiform Encephalopathies(TSE).
Etiology
A transmissible agent that is highly resistant to chemical and
physical agents, and appears not to contain genetic materials(DNA
and/ or RNA).
It is proposed to be due to a prion, a proteinaceous infectious
particle.
The main constituent of these is a disease specific, protease-
resistant neuronal membrane glycoprotein termed the prion 1357
 Scrapie Cont’d---
protein or PrPSC.
PrPSC is an abnormal isoform of a host-coded membrane
glycoprotein PrPC .
The transmissible spongiform encephalopathies are characterized
by the accumulation of PrPSC in neuronal and other tissue.
More than 20 different strains of scrapie have been identified based
on:_
 Strain typing by differences in incubation time in inbred strains of
mice of different genotype.
The type, pattern, severity, and distribution of lesions in the brain of
the different strains of experimental animals (lesion profiles)
Resistance to thermal inactivation
 The type of disease produced in sheep and experimental animals
(e.g. drowsy versus pruritic manifestations in goats)
The ability of a strain to produce disease in different species of
experimental animals.
Co-infection can occur with scrapie. 1358
 Scrapie Cont’d---
But since one will have a shorter incubation period, it will
predominate and be expressed first.
The masked strains can be revealed by passage in different mice
strains.
Epidemiology
Scrapie in sheep occurs enzootically in the United Kingdom,
Europe, and North America.
Outbreaks have been reported in Australia, New Zealand, India, the
Middle East, Japan, and Scandinavia, principally in sheep imported
from enzootic areas.
Under natural conditions, scrapie occurs in sheep and occasionally
spontaneously in goats.
Scrapie is recorded in most breeds of sheep.
But there are breed, family and individual differences in
susceptibility.
There is substantial genetic control of the incidence of disease, and
in both the natural and experimental disease. 1359
 Scrapie Cont’d---
Genetics is a major determinant of susceptibility.
The susceptibility of sheep strongly linked to certain
polymorphisms in the sheep PrP gene.
That is why the incidence of scrapie is higher in some breeds than
others.
For example Suffolk breed in USA and Hill breeds in the UK are
more susceptible to scrapie than other breeds of sheep.
Scrapie is a disease of mature sheep.
Naturally scrapie affects goats very rarely.
However, most of sheep and goats are exposed when they were
young .
The incidence of scrapie decreases with age at exposure.
The age-specific incidence in sheep is highest between 2.5 and
4.5 years of age.
However, scrapie cases are rarely occur under 18 months of age
The case fatality rate, with time in both sheep and goat reaches 100
%. 1360
 Scrapie Cont’d---
Scrapie occurs in both sexes.
The usual method of introduction into unaffected flocks is by the
purchase of pre-clinically infected sheep.
Infectivity can be demonstrated in the placenta and fetal fluids of
naturally occurring cases.
But it has not been demonstrated in the saliva, colostrum, milk,
urine or feces of natural cases, even though it can be demonstrated
in the intestine and nasal mucous membrane.
So that the sources of scrapie prion can be contaminated feed,
more likely contamination of pastures by placentas and fetal
tissues.
Ingestion of infected material appears the most likely route of
infection.
But scarification of the skin and conjunctival inoculation will also
allow infection.
Hay mites have been found to harbor the agent on scrapie infected
properties. 1361
 Scrapie Cont’d---
Generally the methods of transmission of the disease are classified
into
1. Horizontal transmission
2. Vertical transmission
3. Iatrogenic transmission
1. Horizontal transmission
This is the usual method of spreading of the agent.
The placenta is considered the major source of infection for the
mother to her lamb and to other lambs in close contact
Scrapie can also transmit between sheep in close contact.
This can occur from sheep in the preclinical phase of the disease.
Scrapie has also been observed to spread from sheep to goats by
contact.
The duration of infectivity on inanimate materials such as pasture
has not been defined.
But field and experimental observations indicate that it is a long
time, probably in excess of 3 years. 1362
 Scrapie Cont’d---
2. Vertical transmission
There is a greater risk for scrapie in lambs born from infected dams
But this most probably reflects horizontal transmission at birth
from placentas.
There are conflicting results between studies that have examined
transmission by embryo transfer.
That is why the importance of vertical transmission to the
epidemiology of the natural disease remains to be determined.
However, epidemiological studies suggest that it is of rare
occurrence.
Although there is significant evidence against the occurrence of in-
utero - transmission.
But the agent has not been demonstrated in the testes or semen of
rams.
3. Iatrogenic transmission
Many an outbreak of scrapie occurred spontaneously in many
countries following the use of a vaccine against louping ill(ovine
1363
 Scrapie Cont’d---
encephalomyelitis, infectious encephalomyelitis of sheep,
trembling)prepared from the brains of sheep.
More recently, the use of a vaccine against contagious agalactia has
been epidemiologically linked to an outbreak of scrapie in sheep
and goats in many developed countries.
Risk factors
The most common risk factors for the occurrence of scrapie are
1. Environmental and managemental risk factors
2. Animal risk factors
3. Pathogen risk factors
1. Environmental and managemental risk factors
There is a dose-response relationship in naturally occurring scrapie.
The high incidence in some flocks is attributed to a high level of
exposure.
Because of a long winter housing period with a higher risk for
disease in lambs born in the winter housing period.
Factors that influence exposure risk will vary with the management
1364
 Scrapie Cont’d---
systems.
Risk factors that have been identified are
 A higher risk for scrapie in larger flocks and in pedigree flocks
 A greater risk in flocks that lamb communally in group pens
compared to those that in those that lamb in individual pens or
outside on pasture
 A greater risk in flocks that disposed of the placenta in the compost
and spread sheep compost on the land
 A greater risk in flocks that purchased replacement sheep through
the market
 A greater risk where different flocks share pastures or rams.
2. Animal risk factors
We have discussed that scrapie susceptibility difference with breed.
Scrapie in goats is rare and most cases arise in goats that are in
close contact with infected sheep.
Scrapie can spread from goat to goat with no sheep contact.
1365
 Scrapie Cont’d---
Lambs exposed at birth have a shorter incubation period and higher
risk for scrapie than lambs exposed at 6-9 months of age
Similarly, lambs or goats removed from infected dams at birth to a
scrapie free environment have a lower incidence of scrapie than
those removed at later times.
Lambs born to affected ewes are at increased risk for scrapie, and
the offspring from an infected ewe and an infected ram are at
greater risk than those born from an infected ewe and an uninfected
ram.
3. Pathogen risk factors
The scrapie agent can be maintained in tissue culture, and
infectivity is retained with passage.
Infectivity also survives for remarkable periods in dead and
formalinized tissues and infected brain homogenates buried in soil
for 3 years retain their infectivity.
It is highly resistant to physical and chemical influences and can
survive decontamination processes that are effective against 1366
 Scrapie Cont’d---
conventional viruses.
It is capable of withstanding the usual verucidal procedures.
It is not destroyed by boiling, by rapid freezing and thawing, or by
exposure to ether or 20% formalin.
Conventional heat treatments may reduce infectivity
But the agent is remarkable resistant to heat and steam sterilization
at 132°C for 15 minutes is required to totally to destroy it.
Chemical inactivation can be achieved with
 Sodium hypochlorite providing 2% (20000 PPM) of available
chlorine acting for 1 hour,
 4% sodium hydroxide(NaOH)
Economic importance of scrapie
The economic importance of scrapie comprises
Direct loss from the death of animals.
 Curtail the sale of sheep by the public if it is present.
Economic loss for eradication schemes.
Embargos maintained by several countries against sheep from 1367
 Scrapie Cont’d---
enzootic areas.
All this results the dissolution of the pedigree flock as the disease
affects highly pedigree flocks.
Zoonotic implications
There is no evidence for transmission of scrapie to humans or if
scrapie has a risk to public health.
However the concern is that BSE might be established in sheep as a
result of feeding of infected meat-and-bone meal of cattle that are
infected by BSE agent.
This has led some countries to establish an active TSE surveillance
program in sheep.
All sheep that are over 18 months of age and intended for human
consumption are tested at slaughter by a rapid test for scrapie and
BSE.
Pathogenesis
Agent of scrapie shows the predilection sites for tissues of the
lympho reticular system. 1368
 Scrapie Cont’d---
Lympho reticular system is the tissue where the scrapie agent
replicates during the incubation period before invading the nervous
system.
In naturally infected sheep, replication begins in the tonsil,
retropharyngeal and Peyer's patches and gut-associated
lymphoid tissue which probably reflects the oral route of
infection.
However, how the scrapie agent reaches the central nervous system
is not certain.
But it is probably through infection of the autonomic nervous
system.
Gut-associated lymphoid nodules in the Peyer's patches have a
substantial network of nerve fibers and they are probably the site
for neuroinvasion of PrPSC.
Infection in the brain of sheep is initially in the diencephalon and
medulla oblongata with subsequent spread and replication in
other areas of the brain. 1369
 Scrapie Cont’d---
Characteristically, there is a non -inflammatory‘ vacuolar
degeneration of gray matter and the presence of PrPSC in scrapie-
associated fibrils.
Infection results in the post- translational modification of this protein
So that it becomes resistant to proteinases and to normal clearance
and consequently accumulates in the cell.
PrPSC is present not in all placentas and it is present not in all
trophoblast cells of the placentome.
But it is not present in the endometrium, myometrium, associated
nerve plexuses nor in fetus.
The presence of PrPSC in the placenta is determined by the fetal PrP
gene.
PrPSC is not present in the placenta of fetuses carrying resistant
genotypes.
Clinical findings
The incubation period varies from several months upto several
1370
 Scrapie Cont’d---
Years.
Scrapie is a non - febrile disease and the onset is insidious.
But as the disease progresses clinical signs become more obvious
and severe.
The clinical course is protracted, varying from 2 to 12 months but
lasting in most cases for about 6 months.
Affected animals usually show behavioral change which is
manifested clinically by
Tremor
Pruritus
 Locomotary disorder
Scrapie in sheep
There are two stages in the development of scrapie in sheep.
These are
1. Early stage of development
2. Advanced stage of development
1371
 Scrapie Cont’d---
1. Early stage of scrapie
The earliest signs are transient, nervous phenomena occurring at
intervals of several weeks or under conditions of stress.
These episodes include:-
Sudden collapse
Sudden changes of behavior with sheep charging at dogs or closed
gates.
Infrequent occurrence of rubbing and biting at the fleece.
The apparent pruritus is manifested chiefly over the rump, thighs,
and tail base.
The apparent pruritus can be rarely observed in the dorsum of the
neck and less commonly on the neck in front of the shoulder and
the ribs behind the elbow.
The affected areas have approximate bilateral symmetry.
In this early stage a stilted gait(stiff, jerking gait which is caused
by the non- flexing joints of the body) is often observed.
A general loss of condition may also be observed as an early sign 1372
 Scrapie Cont’d---
although the appetite may not be severely affected.
2. Advanced stage of development
It is more advanced stage of the case and shows
 Intense pruritus
 Muscle tremor
Marked abnormalities of gait with severe emaciation
Persistent rubbing causes loss of wool over the areas around rump,
thighs and tail base. dorsum of the neck, the neck in front of the
shoulder and the ribs behind the elbow.
Scratching with the hind feet and biting at the extremities also
occur.
Hematoma of the ears and swelling of the face may result from
rubbing.
Simultaneously with the development of pruritus there is serious
impairment of locomotion which is manifested by
Hind limb abnormalities appear first
There is incomplete flexion of the hock shortening of the step 1373
 Scrapie Cont’d---
weakness and lack of balance.
When the animal is attempting to evade capture gross, there is
incoordination of head and leg movements is likely and the animal
often falls.
Convulsions usually transient but occasionally fatal, may occur at
this time.
General hyperexcitability is evident.
In the animal at rest an intermittent nodding and jerking of the head
and fine tremor of superficial muscles may also be observed.
In some cases, nystagmus can be produced by rotating the head
sideways.
Other clinical signs include
Inability to swallow even if prehension is unaffected
Vomiting
Loss of bleat and blindness
Change of voice
Anorexia is not evident in most cases 1374
 Scrapie Cont’d---
Pregnancy toxemia may occur as a complication in pregnant ewes
during this stage of scrapie.
Finally, the sheep reaches a stage of extreme emaciation
 Inability to move without becoming readily fatigued.
Sternal recumbency follows and lateral recumbency with
hyperextension of the limbs is the final stage.
But pyrexia is not evident at any time
Scrapie in goats
The clinical course in naturally occurring cases lasts from 2 to 24
weeks.
Clinical signs are similar to those in sheep and the common clinical
signs are
Hyperesthesia
Ataxia
Pruritus
But the clinical signs in goat differs from sheep in that loss of
weight is less common. 1375
 Scrapie Cont’d---
In lactating goats the first sign may be a reluctance to permit
milking.
Dribbling and regurgitation of ruminal contents are also recorded in
one-third of cases.
Necropsy findings
Significant gross findings are restricted to traumatic lesions caused
by rubbing and it is loss of wool.
There is gross distension of the abomasum has been recorded in
some natural cases.
The essential histopathological lesion in scrapie is the vacuolation
of gray matter neuropil in the spinal cord, medulla, pons and
midbrain.
Degeneration nerve fibers in the cerebella peduncles and the optic
nerve.
In addition, there is degeneration of the cerebella and
hypothalamus - neurohypophyseal systems.
1376
 Scrapie Cont’d---
Diagnosis
The characteristic signs of behavioral change, tremor, pruritus, and
locomotors disorder occurring during a period of prolonged illness
should suggest the possibility of this disease.
The long incubation period, slow spread and high case fatality rate
should also be considered when making a diagnosis.
There are no changes in hematologic or serum biochemistry
parameters.
Until recently there has been no antemortem laboratory test for
scrapie.
Nowadays, PrPSC can be detected in cells by immuno- histological
methods.
PrPSC is present in the lymphoid tissue of sheep with scrapie in
the preclinical phase of the disease.
However, tonsil biopsy requires general anesthesia and is not a
practical on-farm technique.
Biopsy of lymphoid follicles in the third eyelid is more practical.
1377
 Scrapie Cont’d---
It requires only restraint and local anesthesia and can be a valuable
tool for the preclinical diagnosis of scrapie in surveillance
programs.
In scrapie positive sheep, PrPSC can usually be detected by 14
months.
A sample of a single protuberance on then palpebral side of the
third eyelid will usually give sufficient follicles for analysis.
In laboratory, the agent can be detected by using
 Immunohistochemistry
 Laboratory animals
Because the agent is present in the brain, spinal cord, lymph nodes,
intestinal tract tissue and spleen of infected sheep.
Experimentally, the disease can be transmitted to laboratory
animals like mice; and sheep and goats using extracts from these
tissues.
1378
 Scrapie Cont’d---
Differential diagnosis
Diseases that may require differentiation from scrapie are
1. Diseases with signs of nervous dysfunction including
 Pregnancy toxemia
Rabies
Pseudorabies
Visna
2. Skin diseases like
Different External parasites
Wool loss due to different cases
Treatment
No treatment has proved capable of changing the course of the
disease.
Control and preventation measures
The control and preventation measures of scrapie depends on the
epidemiological situation of the disease in the country as well as in
the farm. 1379
 Scrapie Cont’d---
So that different approaches must b perform to control the disease.
1. If the farm is free from scrapie but the disease is enzootic in
the country the preventation measures must be focused on
individual flocks of the farm.
That means maintenance of a closed ewe flock is critical to control
and prevent this disease.
In the case if ewes and rams need to be purchased from outside this
flock they should be from certified scrapie free flocks or better still,
selected on by PrP genotype testing.
That means the flocks that you have to purchase from breeds that
have resistant gen to scrapie.
In all time the ewes should be isolated at lambing and lambed
individually.
Disposal of placenta must be by burning.
2. In countries that do not have the disease, and where it is
inadvertently introduced with imported sheep, national
eradication program has a good result. 1380
 Scrapie Cont’d---
This approach works on slaughter eradication of the infected flock
and all in-contact animals.
The aim is to eliminate the disease from the country and the
approach is usually successful.
3. In countries where scrapie is enzootic the following programs
play essential role in the control of the disease
1. Flock eradication program
2. Genetic control and national programs
1. Flock eradication program
The eradication of scrapie in countries where it is enzootic has less
chance of success.
The eradication programs vary and may involve the whole flock or
just the family lines of the infected sheep.
During this period there was no antemortem diagnostic test for
scrapie.
The identification of infected farms and flocks relied upon owners
submitting suspect or clinical cases for postmortem and histo 1381
 Scrapie Cont’d---
pathological diagnosis.
To do this the government must prepare adequate compensation for
owners.
The farms are left without sheep for a 2-year period during which
there is extensive cleaning and disinfection of the farm area prior
to repopulation with scrapie-free sheep.
It is the best and effective program to control scrapie in many
countries but very expensive.
2. Genetic control and national programs
The occurrence of scrapie and the concern for BSE in sheep has led
many countries to develop a national breeding programs for the
control of scrapie and potential BSE.
The overall aim is to identify sheep genetically resistant to scrapie
on the basis of their genotype.
Then to breed them so as to create a national flock with scrapie
resistance.
Genetic testing will allow the selection of resistant sheep for 1382
 Scrapie Cont’d---
breeding and the culling of susceptible sheep.
The results obtained by this method suggest eradication is feasible.
But the process could take decades and would be expensive
Surprisingly whole flock culling was more efficient in terms of time
to eradication than genetic typing and selective culling.
NB. The eyelid biopsy test can be used for selected monitoring for
preclinical disease in these flocks.
NB. You must be careful selection for certain PrP genotypes and
reduction or elimination of other PrP genotypes could affect other
desirable genetic characteristics and reduce the overall 'genetic
pool‘ of the country.
So this will need to be determined for individual breeds.
That means preliminary analyses must be done so that we must use
breeds in which reproductive traits, muscle mass, wool quality, live
weight gain, carcass characteristics, is not affected

1383
1384
5.1. Introduction to the order Rickettsiales
5.2. Some common diseases of large animals caused by
Rickettsiales
5.2.1. Cowdriosis(Heart water)
5.2.2. Anaplasmosis

1385
5.1. Introduction to the order Rickettsiales
They are minute, pleomorphic and obligate intracellular Gram –
negative bacteria.
Most of them replicates only in host cells except Rachalimaea
quintana, the pathogenic species of the human being.
They are rods or coccobacilli, visible under the light microscope.
They have the size of 0.3 – 0.6µm in length.
They are aerobic, non – motile and reproduce by binary fission and
budding.
Their reproduction cycle is associated with invertebrate vectors.
Most of them are transmitted by vectors except Coxiella burnetii.
Coxiella burnetii which is the cause of Q – fever in animals and
human beings can be transmitted aerosol through the respiratory
tract or the mucous membranes by infected dust particles and air
droplets.
The invertebrate vectors that are found in phylum arthropods can be
either in class insect( louse and flea) or in class arachnids( ticks and
1386
mites).
 Rickettsiales Cont’d---
The agent can be transmitted in the vectors either transovarially or
transstadially.
Transovarian transmission is characterized by transmission of the
agent to the progeny after the infection of the ovary.
So if the disease is transmitted transovarially to the progeny, the
vectors act as the reservoir of the disease. Example Q – fever in
animals.
In Q – fever the infected animals and ticks are the reservoirs of the
disease.
Transstadially transmission is characterized by the transmission of
the agent through only the stage of developmental cycle ( either
complete or incomplete metamorphosis).
If the agent is transmitted transstadially the vector can not be act as
the reservoir of a disease.
Example human body louse in the case of human typhus which is
caused by Rickettsia prowazenkii.
1387
 Rickettsiales Cont’d---
Rickettsiales infect the vectors either the epithelium of digestive
system or the whole system that lead the accumulation the agent in
ovary and salivary glands.
Vectors are infected by the agent and transmitted it to susceptible
animals while they are sucking the blood of animals as the agent is
located in salivary gland of the vector( most species of Rickettsia).
But some species infest the epithelial cells of the digestive system
of vector and are transmitted to susceptible host by their faces
through micro wounds on skin that are formed during scratching.
Example Human typhus.
Generally the vectors that can transmit rickettsial diseases can be
1. Biological vectors
2. Mechanical vectors
If the agents are reproduced in vectors it is called biological vectors.
If the agents are not reproduce in vectors it is called mechanical
vectors.
There are two families in the order Rickettsiales that have 1388
 Rickettsiales Cont’d---
veterinary importance.
These are
 1. Family Rickettisiaceae
 2. Family Anaplasmataceae
The member of the family Rickettisiaceae of the order have the cell
wall similar to those of other Gram – negative bacteria.
But those belonging to the family Anaplasmataceae have cells
bounded by a two layer of cytoplasmic membrane but lack cell
walls.
Anaplasmataceae and Rickettisiaceae are poorly stain with Gram
stain.
But they stain well with Giemsa and other Romanowsky stains.
So Giemsa or Leishman stain used to demonstrate these organisms
in blood smears.
Only few species of the family Rickettisiaceae will grow in
conventional media. Example Rachalimaea quintana nowadays we
call it Bartonella quintana. 1389
 Rickettsiales Cont’d---
So we can propagate them in the yolk sac of fertile eggs or in
tissue culture.
Others like Haemobartonella felis have not yet been propagated
“in vitro” and “in vivo” in laboratory.
The members of the Rickettisiaceae have tropism for vascular
endothelium or leucocytes.
While the members of Anaplasmataceae parasitize in erythrocytes
and leucocytes.
Most of them are the parasitize in cytoplasm of host cells.
But a few of them replicate in both cytoplasm and nucleus of host
cell. Example spotted fever groups that are caused by R. rickettsii.
Why do the order Rickettsiales are grouped under bacteria but not
protozoa?
Because they do not have true nucleus(nucleus with out nuclear
membrane)
That why the are prokaryotic not eukaryotic
1390
Some common diseases of large animals caused by order
Rickettsiales
Agent Reserv Parasitized Vector Clinically Name of
oir in/on affected the disease
host
Coxiella bruneti Human Mammary Usually Cattle, Q - fever
, cattle gland and Tick but human
and uterus of inhalation and small
small pregnant cow and ingestion ruminants
rumina are means of Cause
nts and transmission abortion
ticks

Ehrlichia equi Horse Granulocytes Vector un Horses in Equine

in USA and known. Tick USA ehrlichiosis
only monocytes suspected
Ehrlichia ondiri Cattle Granulocytes Tick Cattle Bovine
in East and Ambleoma petechial
Africa monocytes fever
high
lands 1391
 Table Cont’d---
Agent Reservoir Parasitized Vector Clinically Name of the
in/on affecting disease
host
Cowdria Domestic Reticular Tick Cattle and Heart water
ruminantum and wild cells, Ambleoma small (Cowdriosis)
(E. ruminants neutrophiles ruminants
ruminantum) in Africa and vascular
and endothelial
Caribbean cells
Anaplasma Cattle and Inside Ticks and Cattle and Anaplasmosis
marginale other erythrocytes other other ( Gall
ruminants with arthropods ruminants sickness)
in tropical marginale and
and sub – distribution veterinary
tropical instrument
regions s

1392
5.2.1. Cowdriosis(Heart water)
Heart water or cowdriosis is a disease of cattle, sheep, goats and
wild ruminants endemic to Sub-Sahara Africa, Madagascar and
portions of the Caribbean.
The disease is characterized by increase in vascular permeability
which leads to a leakage of fluid from blood vessels.
This leakage of fluid results in edema formation, especially in
Pericardium of the heart
In the lungs
Body cavities
The brain of ruminant animals
In goats, the disease is often characterized by renal ischemia (lack
of blood to the kidneys) and nephrosis (disease of the kidneys).
This irreversible kidney damage may cause death in infected goats.
Etiology
Heart water is caused by obligate intracellular rickettsial organisms
that parasitize initially in macrophages and later in endothelium of
1393
blood vessels.
 Cowdriosis Cont’d---
The rickettsial organism is now classified as Ehrlichia
ruminantum, and was previously known as Cowdria
ruminantum.
Ehrlichia (Cowdria) ruminantum is a Gram negative, intracellular
rickettsial organism in the
 Family of Rickettisiaceae
Genus Ehrlichia( formerly called Cowdria)
The organism is coccoid 0.2 - 0.5 µm in diameter and stains blue
with Giemsa stain.
Cyclical development is believed to take place in intestinal and
salivary epithelia of ticks.
Although strain differences exist, all isolates possess a Major
Antigenic Protein - 1 (MAP - 1) that is used for diagnosis.
However, the antigen cross-reacts with other Ehrlichia spp.,
including Ehrlichia equi, the cause of equine granulocytic
ehrlichiosis.
1394
 Cowdriosis Cont’d---
Epidemiology
Heart water is limited in its occurrence to sub- Saharan Africa,
Madagascar and three Caribbean islands.
It is one of the main causes of death in imported breeds of cattle,
sheep and goats in the endemic area.
In endemic areas, morbidity and mortality rates of heart water are
low.
But the percentage of sera positive titers for heart water could be as
high as 100% in adults depending on the abundance of tick vectors.
Case mortality can be as high as 100% in peracute cases in sheep
and goats
However, the case fatality of heart water in cattle in peracute cases
is as low as 0 - 10% in cattle.
The disease is less severe in indigenous breeds than in exotic
imported breeds.
E. ruminantum is transmitted by
1. Biologically by vectors of tick called genus Amblyomma 1395
 Cowdriosis Cont’d---
 The main species of Amblyomma ticks that play in transmission of
heart water are
A. variegatum also known as the tropical bont tick
A. habraeum also called the bont tick
2.Vertical transmission (from cow to calf) through colostrum has
also been reported.
Infection in ticks is transmitted transstadially and possibly
transovarially transmission.
In many books it is written that the organism does not infect
humans.
But researchers in South Africa have recently obtained evidence that
the E. ruminantum may be capable of causing infection in dogs and
people (zoonotic).
It is feared that infected people or dogs may lead to the spread of
heart water to other parts of the globe.
Several wild ruminants can be infected and become subclinical
carriers and reservoirs. 1396
 Cowdriosis Cont’d---
Ticks feeding on them can transmit the disease to domestic
ruminants.
Risk factors for occurrence of heart water
The risk factors are
1. Animal risk factor
2. Environmental risk factor
1.Animal risk factor
Animals at greatest risk are exotics imported into endemic areas.
Native breeds of cattle and wildlife such as the N’Dama breed in
West Africa are said to be resistant to the clinical effects of heart
water.
Angora goats are highly susceptible.
Cattle and sheep recovering from the disease are immune for 6
months to 4 years.
But they can be carriers for 8 months or longer.
Apparently an age-dependent resistance has long been
demonstrated, recognizing that young animals have an innate 1397
 Cowdriosis Cont’d---
resistance.
This resistance has been shown to be due to a low-grade infection
of E. ruminantum obtained in colostral cells.
2. Environmental risk factor
Animals are at greater risk to heart water infection when the vector
population is high, usually during warm and rainy seasons.
Heat water requires the vector tick to get established in any
population of animals where the disease is free.
That means it has the potential to spread from sub Saharan Africa
and Caribbean region etc. to any place where the disease is free if
the vector is present there.
For example heart water can be a spreaded from the Caribbean to
the American mainland if an appropriate biosecurity measures are
not taken.
Therefore, there is concern about possible illegal importation of
infected animals or ticks to southern United States where potential
vectors exist. 1398
 Cowdriosis Cont’d---
Economic importance
Heart water is the most important rickettsial infection of ruminants
in Africa.
It is the second most important tick-borne disease after East Coast
fever.
In southern Africa, it is regarded as the most important disease of
ruminants.
In countries or regions where there is endemic stability, losses from
heart water are minimal until new animals are introduced.
On the other hand, since most losses are in exotic animals, heart
water is a major constraint to livestock improvement in sub-
Saharan Africa in addition of the direct losses by the disease.
Pathogenesis
The rickettsial organisms are introduced into the susceptible
ruminants together with the saliva of an infected tick.
Then after it parasitizes first in macrophages and multiply there.
1399
 Cowdriosis Cont’d---
The organism then infest the reticulo-endothelial cells of the local
lymph node, and rupture the cells and are released into the
circulation.
From where they invade endothelial cells of blood vessels in all
organs.
Organisms can be also found in phagosome of circulating
neutrophils.
But they are more abundant in endothelial cells of blood vessels of
different organs.
They cause increased vascular permeability, leading to edema
especially in the lungs, body cavities and the brain by mechanisms
that are not well understood.
Because the infected endothelial cells show minimal cytopathic
effects.
In goats, renal ischemia and nephrosis have been described during
heart water.
It causes irreversible kidney damage in goats and may be it is the
1400
 Cowdriosis Cont’d---
the cause of death in goats.
Clinical findings
The incubation period is 1-3 weeks after transmission by an
infected tick saliva.
We have discussed that clinical disease is produced by an increase
in vascular permeability .
This phenomenon leads to a leakage of fluid from blood vessels.
This leakage of fluid results in edema formation, especially in the
lungs, body cavities, pericardium and the brain.
In addition to the above features of the disease in goats the bacteria
causes ischemia and necrosis of kidneys.
This irreversible kidney damage may cause death in infected goats.
Depending on the susceptibility of individual animals and the
virulence of the infecting organism, heart water of ruminants may
have the following forms
 Peracute
Acute 1401
 Cowdriosis Cont’d---
Peracute form of cowdriosis is characterized by
High fever that quickly leads to prostration (extreme exhaustion)
Death within 1- 2 days
These animals may have terminal convulsions shortly before death.
The mortality rate may be near 100%.
Acute form of cowdriosis more common and have a course of
about 6 days with the following clinical signs:
A sudden febrile reaction
 In appetence
Listlessness
 Rapid breathing
Classical nervous syndrome which is characteristic of heart water
The affected cattle in acute form of heart water may become
Ataxic
Have empty chewing movements
Twitching of the eyelids
Circling 1402
 Cowdriosis Cont’d---
 Aggression
Apparent blindness and convulsions
The animals will often become recumbent shortly before death.
A profuse, fetid diarrhea often accompanies the neurologic clinical
signs.
The mortality rate is 50 to 90% in adult animals
Whereas calves less than 4 weeks of age only have a 5 to 10%
mortality rate
The mild form of cowdriosis is often subclinical and is seen
mainly in indigenous animals and wild ruminants with high natural
or induced resistance.
Subacute cases have similar clinical signs but they are less severe
than acute form.
The course of the disease in subacute form is longer than acute
form; and it may reach upto 2 weeks in length.
Most animals recover in subacute form cowdriosis.
The mild form of the disease is only seen in indigenous breeds of1403
 Cowdriosis Cont’d---
cattle or native wildlife that have natural resistance.
Necropsy findings
The standard lesions in cowdriosis of ruminants are
 Ascites
 Hydrothorax
 Hydro pericardium.
Pulmonary edema which is often severe and accompanied by
copious froth in the tracheobronchial airways.
There may be subserosal hemorrhages in most cavities.
Lymph nodes are swollen and wet
Spleen is markedly enlarged
Rarely hemorrhages have been described in the brain but often has
no remarkable macroscopically gross lesions
 Microscopically, there is perivascular mononuclear infiltration and
edema along with presence of rickettsial colonies in capillary
endothelial cells.
1404
 Cowdriosis Cont’d---
NB. In goats in addition to the above pathological changes there is
nephrosis and the kidneys are enlarged.
Diagnosis
Diagnose of heart water in ruminant animals can be done by
considering :-
The epidemiology of the disease
The clinical signs of the disease
The necropsy finding of the disease
The result of laboratory finding
Epidemiology, clinical signs and necropsy findings give the
tentative diagnosis of the disease.
However, laboratory finding give the confirmatory diagnosis of the
disease.
In the laboratory findings, hematological changes during heart
water are not specific.
But there may be:-
Thrombocytopenia 1405
 Cowdriosis Cont’d---
Neutropenia
Eosinopenia
Lymphocytosis
Confirmatory diagnosis is done based on the result of
1. Direct microscopy
2. Serological investigation
3. Molecular technique
1. Direct microscopy
Direct microscopy is based on the identification of the rickettsia in
capillary endothelial cells using a Giemsa stained squash
preparation of brain tissue at postmortem.
The rickettsiae occur as blue to reddish-purple colonies or morula
of five to several hundred coccoid organisms (0.2 - 0.5 microns in
diameter) in the cytoplasm of the cells.
Animmunohistochemical staining technique has also been
described.
1406
 Cowdriosis Cont’d---
2. Serological
All serological assays so far available have poor sensitivity or
specificity.
The available serological tests to diagnose heart water are
 An indirect fluorescent antibody test (Indirect FAT) used for
surveys but the close antigenic relationship with other Ehrlichia
spp. often leads to false positives test result that why it is not
widely used nowadays.
 An ELISA based on recombinant MAPl(Major Antigenic Protein –
1) of C. ruminantum was reported to be more sensitivity assay.
It has therefore been suggested as the method of choice for
detection of E. ruminantum infection.
3. Molecular technique
A polymerase chain reaction assay has therefore been suggested as
the method of choice for detection of E. ruminantum infection.
Differential diagnosis
In cowdriosis endemic areas, heart water should be suspected in1407
 Cowdriosis Cont’d---
susceptible animals if
The animals are infected with Amblyomma tick
The animal have a fever of unknown origin
 The above clinical signs are accompanied by nervous signs
The clinical and pathological findings are not specific and the
diagnosis must be based on detection of rickettsial organisms.
The peracute form of cowdriosis should be differentiated from
 Anthrax
 Acute form of rabies
 Sporadic bovine encephalomyelitis
 Tetanus
 Cerebral forms of theileriosis
 Babesiosis
 Trypanosomosis
 Meningitis
 Listeric or other encephalitis
 Hypo magnesemia and poisoning with strychnine, lead and other 1408
 Cowdriosis Cont’d---
organophosphates.
So appropriate laboratory tests must be used to differentiate them.
Treatment
Field cases of heart water are difficult to treat successfully .
Because available drugs are effective only in early febrile stages
before neurological signs develop.
In the early stages, short-acting tetracyclines at 10-20 mg/kg BW
IV and long-acting forms at reduced doses are effective.
Sulphonamides can also be used in the early stages but are less
effective.
Supportive therapy must given to the animal which are useful: -
 In reduction of the pulmonary and other edemas(anti edematic)
In stabilizing the neurologic signs(sedatives)
Control and preventation measures
Control and preventation measures of cowdriosis in cowdriosis
endemic areas can be achieved by having
 1. Appropriate tick control measures 1409
 Cowdriosis Cont’d---
 2. Vaccination programmes
1. Tick control measures
Past efforts to control heart water were based on intensive acaricide
treatment in endemic areas.
It involved frequent use of acaricides (plunge dipping) up to 52
times a year.
This has now been shown to be environmentally unfriendly,
economically unsustainable and would invariably lead to animals
that remained always susceptible.
Nowadays, tick control may be useful or controversial.
So far the use of acaricides have met with only limited success.
For example the use of 1% flumenthrin pour on or plunge dipping
at 45 days interval was found to provide effective protection of
Friesian/Zebu crossbred cattle against important ticks in many
countries of Africa.
However, local breeds like pure Zebu and N'Dama cattle would
probably require less frequent applications of 1% flumenthrin. 1410
 Cowdriosis Cont’d---
But it must be applied correctly at the recommended dose.
In another way the use of acaricide in control of many tick born
diseases in endemic areas is controversial.
Because stringent tick control may result in additional losses by
interfering with immunity obtained when native species of ruminants
are constantly exposed to lower levels of the rickettsial infection.
So that many research works showed in endemic areas, animals may
be better able to deal with field exposure by constant challenge with
low levels of infection.
2. Vaccination
What is advocated today is integrated control based on the
establishment of endemic stability by vaccination or natural
challenge.
More recently, cattle were successfully immunized for upto 10
months of age with
Killed vaccine from a lysate of E. ruminantum formulated in
Freund's adjuvant 1411
 Cowdriosis Cont’d---
Inactivated vaccines from cell-cultured E. ruminantum combined
with an adjuvant led to a reduction in mortality from heart water.
DNA recombinant vaccines
However, as of yet there is no widely effective or safe vaccine
available against E. ruminantum.
Various types of vaccines are currently in the experimental
development stage but have yet to be released.
Chemoprophylaxis involves administration of tetracyclines or
subcutaneous implantation of doxycycline at prophylaxis dose in
susceptible animals when they are introduced into an endemic area
is very important to control and prevent cowdriosis..
However, the results are not always predictable.
NB. In cowdriosis endemic areas the use of cowdriosis resistant
breeds than exotic and cross breeds is useful.

1412
5.2.2. Anaplasmosis
Anaplasmosis has another name called haemo rickettsiasis.
Because the bacteria parasitize in blood cells called erythrocytes.
It is bacterial transmissive diseases of large and small domestic and
wild ruminants caused by intra - erythrocytic parasites characterized
by its acute and chronic form and has the clinical signs like
Intermittent fever which can reach upto 40 – 41 0C
Anemia of visible mucous membrane
 progressive emaciation
Etiology
Anaplasmosis is caused by obligate intra – erythrocytic parasites
belonging to
The order Rickettsiales
Family Anaplasmataceae
Genus Anaplasma
Genus Anaplasma comprises many different species including:-
Anaplasma marginale now it includes A. bovis, A. platys and
1413
 Anaplasma Cont’d---
A. phagocytophilum.
They were previously known as Ehrlichia bovis, E. platys and E.
phagocytophila respectively .
Anaplasma marginale is the causative agent of anaplasmosis in
cattle and wild large ruminants.
Anaplasma ovis is the causative agent of anaplasmosis in sheep
and goats.
Anaplasma centrale is closely related to A. marginale and causes
mild anaplasmosis in cattle.
Anaplasma marginale was originally isolated in Africa.
But it has been introduced as an immunizing agent in Australia,
South America and Asia.
There are antigenic variants among isolates of A. marginale.
The six major surface protein antigens being antigenically
polymorphic.
In blood smears stained by Romancky, Anaplasmas have a coccus
form with the size of 0.2 – 1.2 µm in diameter. and are located 1414
 Anaplasma Cont’d---
marginally in red blood cells.
In blood smears stained by Romancky, anaplasmas organism has a
coccus form with the size of 0.2 – 1.2 µm in diameter.
They are located marginally in red blood cells.
Very rarely anaplasmas can be found in leucocytes and
thrombocytes of affected animals.
A. marginale retain red color in basic fuchsine but blue in
methylene blue stainings.
Anaplasmas are the parasites of red blood cells of large and small
ruminants.
They reproduces by binary vision and budding .
During reproduction the anaplasmas are found in erythrocytes by
forming 2 - 8 anaplasmas cells in group.
This groups of cells in cytoplasm of erythrocytes are surrounded
by two layers of plasma membrane are called microcolonies.
Microcolonies of anaplasmas are clearly visible under
electronmicroscopy. 1415
 Anaplasma Cont’d---
Epidemiology
Anaplasmosis in cattle is common world wide.
It is transmitted by a diverse group of biological and mechanical
vectors.
Infection in cattle is endemic in tropical and subtropical areas.
Because the environment supports these vectors to be found there
constantly.
Infection occurs more sporadically in temperate climate areas.
Anaplasmosis of sheep and goats has a distribution similar to that
of cattle
Cattle are susceptible to
A. marginale and
 A. centrale
Sheep and goats are susceptible A. ovis.
A variety of species of wild ruminants in both North America and
Africa can be infected and may have significance as reservoirs for
A. marginale. 1416
 Anaplasma Cont’d---
For example black-tail deer in west cost region of USA and
different species of antelope in south Africa are the main reservoirs
of the disease.
Bighorn sheep may be reservoir for A. ovis in western USA.
The prevalence of infection in cattle in endemic areas is very high
with seropositivity rates exceeding 60% and often approaching
90% .
Differences in seropositivity of cowdriosis in enzootic and
epizootic areas are largely related to tick distribution and climate.
However, there is a concern, and some evidence, that the global
warming trend will expand the boundaries and movement of host
ticks.
The source of infection is always the blood of an infected animal.
Recovery from acute infection results in persistent infection.
Persistent infection is characterized by repetitive cycles of
rickettsemia.
Persistent carriers are the reservoir for herd infection. 1417
 Anaplasma Cont’d---
However, the level of rickettsemia is often too low for detection by
microscopy.
But it can be detected by nucleic acid probe analysis.
Generally transmission of Anaplasmas can be classified as
1. Haematophages insect transmission
2. Iatrogenic transmission
3. Transplacental transmission
1. Haematophages insect transmission
It is the transmission of anaplasmas from one animal to another
animal chiefly by insect vectors.
A variety of arthropods may act as vectors.
But significant natural vectors are
Ticks in the family Ixodidae and
 Flies in the family Tabanidae.
It is known that over 20 species of tick have been incriminated a
vectors world-wide.
1418
Among all these ticks , the most importants are
 Anaplasma Cont’d---
The one-host Boophilus spp. are of major importance in tropical and
subtropical regions
 The three-host tick Dermacentor spp. of major importance in the
western USA
The organism undergoes a complex developmental cycle in the gut
cells of ticks
The final infective stage is present in the salivary gland of the ticks
In ticks there are two methods of transmission of anaplasmas.
These are
Transovarial(rarely)
Transstadial( usually)
In ticks anaplasmas develop in gut and move to salivary glands and
ticks are biological vectors of anaplasmas.
There appears to be no developmental sequence of Anaplasma spp.
in flying insects.
So that tabanids are efficient mechanical vectors but not biological
1419
vectors.
 Anaplasma Cont’d---
They can transmit infection for 2 hours after feeding.
2. Iatrogenic transmission
Anaplasmosis may also be spread mechanically by different fomites
Infected hypodermic needles
 Castrating, spaying and dehorning instruments
 Embryo transplants(infected embryo and/ or instrument)
Blood transfusions(infected blood and/ or instrument)
NB. Anaplasmosis may also be spread when cattle, used as donors
of infected blood for immunization against babesiosis when there is
mixed infection with A. marginale.
3. Transplacental transmission
Intra-uterine infection also occurs in cattle.
But much less frequently in field cases than in experimental ones.
Abortion or neonatal infection may result.
Risk factors for occurrence of anaplasmosis
As many diseases of animals, risk factors for occurrence of
anaplasmosis can be grouped in to three groups. 1420
 Anaplasma Cont’d---
These are
1. Animal risk factors
2. Environmental risk factors
3. Pathogen risk factors
1. Animal risk factors
The animal risk factors are
Breed
Nutritional status
Age at infection
Bos indicus, Bos taurus and their crosses have equal susceptibility
to anaplasmas infection.
But under field conditions, Bos indicus are not as commonly
affected as compared to Bos taurus.
Because Bos indicus has relative resistance to heavy tick
infestation.
Breeds with black or red coat color have a higher risk of infection
than those with white coats in regions where biting flies in the 1421
 Anaplasma Cont’d---
family Tabanidae. are the insect vector.
Dairy breeds may be at greater risk for iatrogenic transmission than
feedlots.
Exposure of infected but clinically normal animals to devitalizing
environmental influences like shortage of feed and the presence of
other concurrent diseases may result in the development of acute
anaplasmosis.
Age at infection is a major determinant of the severity of clinical
disease of young animals.
Young calves are less susceptible to infection with A. marginale
than older cattle although the reason for this is not understood.
Infection between six months and three years of age has increasing
risk of clinical illness.
Animals infected after 3 years of age are commonly affected by a
peracute fatal form of the disease.
1422
 Anaplasma Cont’d---
2. Environmental risk factors
Environmental risk factors in the occurrence of anaplasmosis are
Season of the year
Geographical location
In tropics the disease may occur in any season of the year.
But the outbreaks of anaplasmosis is more correlates with warm
and rainy season of the year.
In temperate climates a seasonal occurrence of disease occurs in
association with seasonal occurrence of the insect vectors.
Winter outbreaks are likely associated with iatrogenic
transmission or possibly the winter tick called Dermacentor
albipictus.
Clinical disease is rare in enzootic areas.
Because the infection pressure is high and most cattle may develop
immunity after clinical and subclinical infection.
However, the animals become susceptible to anaplasmosis in
anaplasmosis when they are seronegative to anaplasmas infection.
1423
.
 Anaplasma Cont’d---
Clinical disease of anaplasmosis occurs where there is
Introduction of susceptible animals into endemic areas and/ or
The expansion of the vector population into previously free areas
and/ or
When there is interface between endemic and non- endemic
regions.
3. Pathogen risk factors
Phylogenetic analysis of A. marginale geographic isolates support
the existence of clades.
Australian isolates do not appear to differ significantly in
antigenicity or virulence.
In contrast in other countries there can be significant differences
between isolates in antigenic composition.
Recent research has demonstrated that the phenomenon of
infection exclusion occurs with A. marginale.
Infection of tick cells and bovine erythrocytes with one genotype
of Anaplasma marginale excluded infection with other genotypes. 1424
 Anaplasma Cont’d---
So that in herds of cattle from endemic areas where many
genotypes were detected only one genotype was found per animal.
Economic importance
The economic loss due to anaplasmosis comprises from
 Direct death and abortion in clinical cases
 Loss of production in sick and recovered animals
Economic loss associated with preventive measures such as tick
control and medication of clinically sick animals.
Difficulty in exporting of cattle to countries where the disease is
not common( developed world).
Limitation to the introduction of susceptible cattle breeds with
superior genetics(developing world).
Pathogenesis
Anaplasma are obligate intra-erythrocytic bacteria.
They infect mature erythrocytes by an endocytic process and
reproduction occurs by binary fission to produce 2 - 8 infective
initial bodies which are covered by plasma membrane in cytoplasm
1425
 Anaplasma Cont’d---
of infected erythrocytes called microcolonies.
After reproduction, Anaplasma. marginale leave the infected
erythrocytes by exocytosis and other erythrocytes.
The number of infected erythrocytes doubles every 24 - 48 hours
and the infection becomes patent 2 - 6 weeks after infection.
The length of time when the clinical signs become patent depends
on
 The initial challenge dose of A. marginale
 The strain of the pathogen
The susceptibility of the host
Upto 10 - 90% of erythrocytes may be parasitized in the acute stage
of the infection.
However, at least 15% of erythrocytes have to be parasitized to
develop clinical disease of anaplasmosis.
Parasitized erythrocytes are removed by phagocytosis in the
reticular endothelial system.
1426
 Anaplasma Cont’d---
This results in release of acute phase inflammatory reactants and
the consequent development of fever.
Continued erythrocyte destruction resulting in the development of
Mild to severe anemia
Icterus without hemoglobinemia and haemoglobinuria
The first appearance of the anaplasmas in the blood coincides with
 A fall in the hematocrit (reduction of PCV value)
 Reduction of erythrocyte numbers
 The appearance of immature erythrocytes in blood smears
 The development of fever.
Acutely affected animals may die shortly after this phase is reached.
The appearance of anti-erythrocyte antibodies late in the acute stage
may exacerbate the anemia.
Cattle that survive the disease become carriers and serve as
reservoirs of A. marginale and have cycles of rickettsemia.
Because they provide a source of infective blood for both
mechanical and biological transmission by ticks. 1427
 Anaplasma Cont’d---
Clinical findings
In cattle, the incubation period varies with the challenge dose and
resistance of animals.
But it is generally about 3 - 4 weeks with tick-borne infection and 2
- 5 weeks with the inoculation of blood.
In cattle, anaplasmosis has the following forms
Per acute
Subacute
Peracute cases of anaplasmosis is characterized by
 A sudden onset of high fever
Anemia followed by icterus
Abortion of pregnant cows
Severe dyspnea
 Hyperexcitable and tending to attack attendants just before death
In most cases, anaplasmosis of cattle has subacute form and
common especially in young animals.
The common clinical features of subacute form of anaplasmosis1428 in
 Anaplasma Cont’d---
cattle are
Rising of body temperature which may reach upto 40.5 0C which
may remain elevated or fluctuate with irregular periods of time.
Jaundiced and marked pallor of the mucous membrane with out
haemoglobinuria
Impairment of fertility
Death
However, many of animals survive in an emaciated condition and
become carriers with impairment of their fertility.
In sheep and goats, anaplasmosis has usually subclinical form.
But in some cases, particularly in goats, a severe anemia may occur
and a clinical picture similar to that found in cattle may be seen.
Goats may show hyperexcitability and may bite at inanimate
objects.
The main clinical pictures in lambs includes
Fever
Anemia 1429
 Anaplasma Cont’d---
Pale and icteric conjunctivae
Constipation or diarrhea
Necropsy findings
The most obvious necropsy findings in anaplasmosis are
Emaciation
Pallor of the tissues
Thin and watery blood
 Mild jaundice
Enlargement of liver which has orange colour
Enlargement of spleen with soft pulp
Congested kidneys
 Myocardial hemorrhages
Reddened of the bone marrow cavity because of the increased
hematopoietic tissues in acute case.
Atrophy of bone marrow in chronic cases
Postmortem identification of A. marginale can be established by
Staining blood smears with Giemsa and/or fluorescent stains. 1430
 Anaplasma Cont’d---
To do this peripheral blood is superior to organ smears.
The technique is applicable to fetuses suspected of being aborted as
a result of infection with Anaplasma spp.
Diagnosis
Diagnose of anaplasmosis can be done based on
 The epidemiology of the disease
 Clinical signs
 Necropsy findings
 Laboratory findings
The common laboratory methods that are used to diagnose
anaplasmosis are
 1. Hematology
 2. Direct microscopy
 3.Isolation and identification of the bacteria(serology and
molecular technology)
 4. Serology
1431
 Anaplasma Cont’d---
Specimens
The specimens of choices for confirmatory diagnosis includes
Blood smears from cut surface of an ear
 Spleen, liver and bone marrow for bacterial isolation and for
histopathology
1. Hematology
Erythrocyte destruction may be so severe that the erythrocyte count
is reduced to 1.5 million/µl
Immature red cells are common at this stage and their presence is
considered to be a favorable sign. To diagnose blood parasitic
bacteria, protozoa and virus.
The small dot-like bacteria are discernible at the periphery of
upto 10% of the red cells in subacute cases.
But in peracute cases more than 50% of the cells may be affected
by the agent.
A. ovis are usually situated at the periphery of erythrocytes but as
many as 40% of infested cells may show sub marginal bacteria.1432
 Anaplasma Cont’d---
2. Direct microscopy
It is used mostly during rickettsemia in peracute and acute cases
The method is based on staining of blood smears with Giemsa
and/or fluorescent stains.
Peripheral blood is superior than others.
The technique is applicable to fetuses suspected of being aborted as
a result of infection with Anaplasma spp.
3. Isolation and identification of the bacteria
The bacteria can be isolated by cultivating it using
Embryonated egg by inoculating the filtrate through the yolk sac
route
Cell cultures
Laboratory animals (mice or guinea – pigs)
Transmission of the filtrate to splenectomized animals that is taken
from latently infected animals has been used to detect carriers.
But it is not widely used because it is expensive.
1433
 Anaplasma Cont’d---
The bacteria can be identified by
Serology
Molecular technique
Direct staining of specimens does not help to identify the species of
the bacterium.
Specimens from sick animals, from egg yolk sac and laboratory
animals can be identified by using FAT( direct FAT).
Nucleic probe analysis can be used to detect low levels of
rickettsemia.
Polymerase Chain Reaction(PCR) is widely used to identify the
bacteria.
4. Serology
There are many serological tests to diagnose the presence of
antibody in infected herds.
The most common and practically applicable serological tests are
Complement Fixation Test(CFT)
Rapid Serum Agglutination Test(RSAT) on the card 1434
 Anaplasma Cont’d---
Capillary tube agglutination test
Indirect Fluorescence Antibody Test(Indirect – FAT)
Indirect ELISA
Competitive ELISA
The complement fixation test is the standard test for the detection of
carrier animals.
It is satisfactory to use in cattle, goats and sheep.
But the antibody titer is highest during the active phase of the
disease and sufficiently low in carrier animals.
So there is some proportion of false negative test results.
False positive reactions can also occur because of erythrocyte
contamination of the A. marginale antigen and the presence of
antibodies to erythrocytes in some cattle sera.
A rapid card agglutination test, which tests serum or plasma for
antibodies against A. marginale is cheap and quick and sufficiently
accurate to be used as a herd test.
So that many countries use RSAT for screening test and CFT to1435
 Anaplasma Cont’d---
confirm the test.
A capillary tube agglutination test is comparable efficient and is
available and more economical and faster.
An indirect fluorescent antibody test is also accurate and has a
particularly suitability for testing blood which has been dried onto
paper for passage through the mails.
ELISA with high sensitivity, specificity and predictive value is also
described and could be particularly applicable to field examinations.
A competitive inhibition ELISA test, with high sensitivity and
specificity has been developed that detects antibody to a major
surface protein that is conserved among anaplasma species
NB. Vaccinated animals may react to all of the serological tests for
periods of over one year and you must consider it as the serological
tests never differentiate antibody from natural infection or after
vaccination
Differential diagnose
Anaplasmosis must be differentiate from many disease of cattle 1436
and
 Anaplasma Cont’d---
small ruminants that cause haemolytic anemia.
Some of these diseases those can be considered as differential
diagnosis are l
Babesiosis
Piroplasmosis
 Anaemia caused by organic and inorganic toxins
Leukosis in cattle
Babesiosis and piroplasmosis can be differentiate from
anaplasmosis in that
Babesiosis and piroplasmosis cause haemoglobinuria but not in
anaplasmosis.
Anaplasmas are located marginally as dot point in erythrocytes
under microscope but the two protozoal parasites have different
morphological features in erythrocytes.
Leukosis in cattle can be differentiated from anaplasmosis in that
Leukosis has hereditary character and is caused by virus
Leukosis does not have seasonal character and does not show 1437
 Anaplasma Cont’d---
treatment effect with tetracycline groups.
Anaemia due to intoxication can be differentiate from anaplasmosis
in that
It does not has seasonal characteristics and does not correlates with
tick population.
Has no as such mass character in given area.
 Absence of anaplasmas in red blood cell of affected animals.
Treatment
Anaplasmosis can be treated with tetracycline groups.
 1.Treatment of clinical disease can be achieved with
oxytetracycline 6-10 mg/kg BWIM daily for three days or
A single injection of long-acting oxytetracycline at a dose of 20
mg/kg IM.
Concurrent administration of estradiol cypionate (14.3 mg/kg BW
intramuscularly) appears to improve the rate of recovery by
promoting parasitemia during treatment.
Tetracycline treatment will not eliminate infection and immunity1438
 Anaplasma Cont’d---
persists for a long time.
Blood transfusions are indicated as supportive treatment in animals
with a PCV less than 15 % .
2. Imidocarb (3 mg/kg BW) is also an effective treatment for
clinical cases and does not interfere with the development of
acquired immunity to A. marginale.
The risk for infection in the rest of the herd should be assessed and
protection with tetracycline must be given to animals in
prophylactic dose.
Protection of animals from anaplasmosis with tetracycline can be
used for
Temporary protection
Prolonged protection
Elimination of infection
Temporary protection in the face of an exposure risk can be
achieved with a single intramuscular injection of long-acting
tetracycline at dose of 20 mg/kg BW. 1439
 Anaplasma Cont’d---
The results generally are good except when the cattle are exposed
to infection during the 14 days prior to the treatment.
Prolonged protection can be achieved by the intramuscular
injection of long-acting tetracycline at the dose of 20 mg/kg BW of
every 28 days or by chlortetracycline in the feed at 1.1 mg/kg BW
daily.
Elimination of infection cannot be achieved with tetracycline
therapy.
Preventation and control measures
Methods for the control of anaplasmosis have not changed greatly
over the past several decades and comprises of the following
methods.
Arthropod control with acaricides
Chemotherapy for prophylactic measures
Vaccination
The eradication of anaplasmosis is not a practicable procedure
in most countries at the present time. 1440
 Anaplasma Cont’d---
This is because of : -
 The wide range of insects which are capable of carrying the
disease
 The long period of infectivity of carrier animals
 In some areas, the presence of carriers in the wild animal
population.
Nowadays, the following methods are commonly used to prevent
and control anaplasmosis.
These are
 Taking of general measures
 Controlling animal movements
 Elimination of carriers
 Controlling of outbreaks
 Using of chemotherapy as prophylaxis measure
 Vaccination
1441
 Anaplasma Cont’d---
Taking of general measures
The introduction of the disease into herds by carrier animals
should be prevented by prior serological testing.
Attention should also be given to preventing iatrogenic
transmission with instruments used for injections or surgical
operations by sterilization(disinfection) after using on each animal.
Controlling animal movements
Some advantage can be gained when introducing animals into an
enzootic area by
Limiting the introductions to animals of less than 2 years of age
Bringing them in when the insect and tick population is least
numerous
Elimination of carriers
This is feasible in regions that are subject to only periodic
incursions of infection and that do not have endemic tick vectors.
It can be achieved by serologic testing and culling of reactors or
treating them as outlined above to eliminate the carrier state. 1442
 Anaplasma Cont’d---
Controlling of outbreaks
If an outbreak occurs:-
The affected animals should be treated vigorously as described
above
 In-contact animals must be vaccinated and/or placed on a regimen
of prolonged tetracycline protection at prophylactic dose.
 Subsequently all exposed animals should be tested serologically
and the reactors treated or preferably salvaged.
Using of chemotherapy as prophylaxis measure
Chemotherapy for control is more commonly used in many
countries.
It is expensive and carries the risk of causing selection of resistant
strains.
Vaccination
Exposure-naive animals that are to be introduced into an enzootic
area should be vaccinated.
Most control programs in enzootic areas are based on increasing 1443
 Anaplasma Cont’d---
the resistance of the population by immunization.
In any vaccination program particular attention should be paid to
the animals at high risk consists of:-
 Animals particularly brought in from non-enzootic areas
Those in surrounding similar areas to which infection may be
spread by expansion of the vector population under the influence of
suitable climatic conditions.
There are two types of vaccine to prevent cowdriosis in cattle and
small ruminants
Killed vaccines
Live attenuated vaccines
Killed A. marginale vaccine are usually in an adjuvant vehicle.
The vaccine requires two doses, four weeks apart, the last dose
given at least two weeks before the vector season.
Subsequently, booster doses should be given two weeks before the
next vector season.
The vaccine does not prevent infection but does significantly
1444
 Anaplasma Cont’d---
The duration of the immunity lasts at least for 5 months.
There is a risk for neonatal isoerythrolysis.
This can be reduced by vaccinating only non- pregnant cows and
avoiding unnecessary booster injections.
Live attenuated vaccines have been attempted by irradiation of
strains and passage of the organism through sheep or deer and the
use of naturally low virulence isolates.
Live vaccines of anaplasmosis have demerits because
 The vaccine is currently produced in live animals which is
expensive.
Since the vaccine is non-inactivated vaccines there is a risk of
transmitting blood-borne viral and protozoal diseases.
Cattle that have recovered from acute infection or immunized with
killed vaccines are solidly protected against challenge with the
homologous strain.
However, they are partially protected against challenge with
hetrologous strains. 1445
1446
6.1. Introduction to fungi
6.2. Dermatomycosis
6.2.1. Ringworm

1447
 Since fungi are larger than bacteria, they were recognized as the
agent of infectious diseases earlier than other infectious agents
(bacteria and virus).
Nowadays, even though there are over 250, 000 fungal species,
only 150 species are pathogen for animals and human beings.
Infectious diseases which are caused by fungi are called mycoses(
singular mycosis).
Fungal infections occur
1. By invasion of tissues, and the diseases are called mycosis.
2. By production of different toxins which are called mycotoxin,
and the diseases that are caused by mycotoxins are called
mycotoxicoses ( singular mycotoxicosis).
3. By induction of hypersensitivity called allergy.
Factors which may predispose animals and human beings to fungal
invasion are:
 Immunosuppression due to various reasons
1448
 Introduction Cont’d---
 Immunosuppressive viral diseases ( HIV/ AIDS) in human being.
Prolonged antibiotic therapy
Immunological defects
Immaturity (ageing and malnutrition)
Exposure to heavy challenges of fungal spores
Traumatized tissue
Persistent moisture to the skin surface etc.
Traditionally, the mycotic diseases are classified according to the
site or in the level of the infected tissues.
Classification of mycotic diseases are mostly followed by
researchers and teachers because it facilitates an easy understanding
for us.
So that clinically all mycotic diseases (mycoses) are classified as
1. Superficial mycoses
2. Cutaneous mycoses
3. Subcutaneous mycoses
1449
4. Systemic mycoses
 Introduction Cont’d---
1. Superficial mycoses
This group of fungi affect only the upper layer of epidermis; the
super follicular layer and are localized along hair shafts of the hair.
Therefore, the infection is localized to the stratum of the outer, non
– viable, cornified epidermal cells.
In hair infections the growth is generally superficial with minimum
damage.
This group of fungi do not infect other part of the body.
They rarely induce the immune response of the infected host.
2. Cutaneous mycoses
This kind of fungal infection is caused by dermatophytes ( Gr.
Phyton which means plant) that are always parasites.
The disease which is caused by dermatophytes is called
Dermatophytosis (plural dermatophytoses) or dermatomycosis
(plural dermatomycoses).
The fungal agents affect the epidermal tissues, hair and claws (nails)
of animals and human beings. 1450
 Introduction Cont’d---
They invade and destroy the keratinized tissues of the body ( skin,
hair, claws/ nail and feather)
As the result they cause mild to severe inflammatory reaction in the
host.
The infection is characterized by the formation of raised, circular
and crusted patches beside alopecia and itches.
This group of fungi do not penetrate the subcutaneous or deeper
tissues of the body.
Because naturally there are some inhibitory factors in the body
fluids and serum.
Incidence of cutaneous form of the disease is more in young age
group than adults.
Because the skin of young is thinnest and smooth.
A good example of this form of fungal infection which is caused by
dermatophytes is ring worm.
3. Subcutaneous mycoses
It is caused by saprobes in soil and on vegetation. 1451
 Introduction Cont’d---
Infection occurs by direct implantation of spores or mycelial
fragments, commonly in scratches caused by thorns.
That is why subcutaneous mycoses are more prevalent in rural and
tropical regions.
Subcutaneous mycoses affect skin, subcutaneous tissue, fascia and
bone.
The diseases begin insidiously, progress slowly and are
characterized by localized subcutaneous abscesses and granulomas
that spread by direct extension.
4. Systemic mycoses
It is also called deep mycoses.
Systemic or deep mycoses are caused by saprobe fungi in soil and /
or opportunistic fungi that are commensals on the skin and mucous
membranes.
The route of infection for most systemic mycoses is inhalation.
Infection of systemic mycoses resemble clinically to the chronic
bacterial infections due to mycobacteria and actinomycetes. 1452
 Introduction Cont’d---
The earliest lesions are usually pulmonary, and it is characterized
by acute, self – limited pneumonia like that of bacteria and virus
infections.
The subsequent chronic form ( which is usually less frequent)
begins insidiously, progress slowly, and is characterized by
suppurative or granulomatous lesions.
These some times form pulmonary cavities and often spread by
direct extension. Example in to contiguous soft tissues such as
pleurae.
Dermatomycoses are caused by dermatophytes fungi that infect
only epidermis and its appendages like hair and claws/ nails of
animals.
Dermatophytes infect structures that are reach in keratin.
This group of fungi have affinity to keratin – rich tissues because
they have unusual capacity to degrade and utilize keratin.
The skin lesions are usually roughly circular, tend to expand
1453
 Introduction Cont’d---
equally in all directions, and have raised serpiginous borders.
Because of the morphology of the lesions on the skin, in ancient
time people thought to be due to worms or lice.
That is why up to know dermatomycoses are called ring worm or
tinea( it is Latin word which means worm or the larva of insect).
The name of ring worms are usually qualified by the area of skin
involved.
Example ring worm around phalanges/ fingers is called Tinea
capitis; ring worm around the body of animals is called Tinea
corporis and ring worm on the feet of human being is called Tinea
pedis.
Some useful characteristics of dermatophytes
All are septate and on - dimorphic fungi (exist only in mycelial
form in tissues and cultures)
Arthrospores are the infectious form of dermatophytes.
They do not have sexual form of reproduction so that they are
grouped under Fungi Imperfect. 1454
 Introduction Cont’d---
Macroconidia of different morphology may be formed on/ in
cultures.
All are obligate aerobes, grow slowly( 14 – 30 days) and
require 25 0C for their growth “in vitro”.
Depending on colonial pigmentation, morphology of spores
and other characteristics like presence or absence of
arthrospores and chlamydospores etc all dermatophytes fall in
to three genera.
Microsporum
Trichophyton
Epidermophyton
The most important genera in veterinary medicine are
Microsporum and Trichophyton.
Genera Epidermophyton is represented by single species E.
floccosum and affects only human being. 1455
 Introduction Cont’d---
Some common species of dermatophytes which causes
dermatomycoses
Dermatophytes Species of animals Geographical distribution
M. equinum Horses Africa, Australia, Europe,
North and South America
M. gypseum Horses, dogs and rodents World wide
M. nanum Pigs North and South America,
Europe and Australia
Trichophyton equinum Horses ( mostly under 4 years World wide
of age)
T. equinum var. Horses Australia and New Zealand
autotrophicum
T. mentagrophytes Rodents, dogs, horses and World wide
var. mentagrophytes many other animal species

T. verrucosum Cattle (mostly calves) World wide

1456
 Introduction Cont’d---
6.2. Dermatomycosis
6.2.1. Ringworm
It is a fungal disease of large animal characterized by invasion of
cutaneous keratinized epithelial cells and hair fibers by
dermatophytes.
Etiology
Ringworm is caused by groups of fungi imperfect belonging to
genus
 Microsporum
 Trichophyton
 Epidermophyton
The most important genera in veterinary medicine are Microsporum
and Trichophyton.
Genera Epidermophyton is represented by single species E.
floccosum and affects only human being.

1457
 Ringworm Cont’d---
Some of the species of genus Microsporum and Trichophyton and
affected species of large animals
Cattle Pigs Sheep Goats
Tr. verrucosum Tr. mentagrophytes Tr. verrucosum var. Tr. verrucosum.
Tr. mentagrophytes Tr. rubrum ochraceum
Tr. megninii Tr. verrucosum Tr. quinclceanum
Tr. verrucosum var. var. discoides, Tr. mentagrophytes
album M. canis, Tr. gypseum
Tr. verrucosum var. M. nanum
disco ides

1458
 Ringworm Cont’d---
Epidemiology of ringworm
Ringworm occurs in all animal species in all countries.
But it is more commonly where animals are accommodated in
dense groups.
Ringworm can be transmitted by
Direct contact with infected animals ( the common method of
spread of ringworm)
 Indirect contact with inanimate objects, particularly bedding,
harness grooming kits and horse blankets
Spores can exist on the skin without causing lesions for a long time
and these groups of animals can act as source of infection.
Premises and harness may remain infective for long periods.
Because fungal spores remain viable for years provided they are
kept dry and cool.
Moderate heat and desiccation destroy them.
Risk factors for occurrence of ringworm
The common risk factors are 1459
 Ringworm Cont’d---
1. Pathogen risk factor.
3. Host risk factor
3. Environmental risk factor
Although most of cutaneous fungi cause mycoses by invasion of
tissues, M. gypseum and M. nanum are soil saprophytes and the
reasons for their assumption of pathogenicity are not understood.
Animal susceptibility is determined largely by immunological status
of the organism.
So that young animals, animals with different abrasions and stressed
animals(nutrition deficiency etc.) are the most susceptible animals
for cutaneous mycoses.
A high incidence of clinical cases in the winter and of spontaneous
recovery in the spring is common.
But outbreaks also occur during the summer months.
So that close confinement and possibly nutrition seem to be more
important in the spread of the disease than other environmental
1460
 Ringworm Cont’d---
factors such as temperature and sunlight.
Humidity is known to be important, a high humidity being
conducive to multiplication of the fungus.
In calf-rearing units the prevalence of ringworm is greater in units
which continuously add or remove calves from the stock;.
So that an 'all-in all-out‘ program is less conducive to control the
spreading of the ringworm.
Zoonotic implication of ringworm
Ringworm of animal origin affects adult humans as well as children.
Diagnosis and treatment of ringworms are often very difficult.
About 80% of human ringworm may derive from animals.
The source of infection for humans can be varied with species of
fungi
 Trichophyton spp. infections are commonly contracted from horses
and cattle
 M. canis infections from dogs.
1461
 Ringworm Cont’d---
Trichophyton verrucosum from cases tinea corporis, tinea faciei
and tinea captis were
Identified in many countries of the world.
Prevalence of ringworm is common in
 Farmers those have contact with animals
 Children who lived on dairy or cattle farms or in a dairy farming
area
Slaughterhouse employee
 Veterinarian
Economic importance of ringworm
Skin injury to affected animals by dermatomycoses is minor
nature.
But sufficient damage to hides occurs to warrant some attempt to
control the disease.
Pathogenesis
Ringworm fungi chiefly attack keratinized tissues particularly the
stratum corneum and hair fibers. 1462
 Ringworm Cont’d---
This results in
Autolysis of the fiber structure
 Breaking off of the hair and alopecia
Exudation from invaded epithelial layers, epithelial debris and
fungal hyphae produce the dry crusts which are characteristic of
the disease.
The lesions progress if suitable environmental conditions for
mycelial growth exist, including a warm humid atmosphere, and a
slightly alkaline PH of the skin.
Ringworm fungi are all strict aerobes and the fungi die out under
the crust in the center of most lesions, leaving only
the periphery active.
It is this mode of growth which produces the centrifugal
progression and the characteristic ring form of the lesions.
The susceptibility of humans to ringworm infection is much
greater before puberty than afterwards. this is because of: -
1463
 Ringworm Cont’d---
This is because of the falling of the PH of skin from about 6.5 to
about 4.0.
This change is largely due to excretion of fatty acids in the sebum
and these fatty acids are often highly fungi static.
Calves are more commonly infected than adult cattle.
But whether this is due to increased susceptibility in calves or the
development of immunity in adults has not been determined.
There is some experimental evidence that traumatic injury of the
skin is an important factor for the development of ringworm
lesions in calves.
Different numbers of macroconidia of Trichophyton verrucosum
are required to induce ringworm depending on the degree of
shearing of the hair and scarification of the skin.
Secondary bacterial invasion of hair follicles is common.
A resistance to reinfection occurs after recovery from experimental
or natural infection.
The immunity is specific to the fungal species concerned and lasts
1464
 Ringworm Cont’d---
upto 2 years.
Clinical findings
The clinical findings are almost similar in different species of large
animals and have the following features.
In cattle
The typical lesion is a heavy, gray-white crust raised perceptibly
above the skin.
The lesions are roughly circular and about 3 cm in diameter.
In the early stages the surface below the crust is moist.
However, in older lesions the scab becomes detached and
pityriasis and alopecia may be the only obvious abnormalities.
Lesions are most commonly found on the neck, head and perineum.
But a general distribution over the entire body may occur
particularly in calves.
In severe cases the lesions may coalesce together.
Itching does not occur and secondary acne is unusual. 1465
 Ringworm Cont’d---
2.In pigs
Regular ringworm lesions in pigs develop as a centrifugally
progressing ring of inflammation surrounding a scabby,
alopecic center.
The lesion produced by M. nanum is different because
There is no pruritus or alopecia
Cutaneous reaction is minimal but the centrifugal enlargement
of each lesion may cause it to reach an enormous size.
Superficial, dry, brown crusts cover the affected area.
But they are not obviously raised except at the edges in some
cases.
The crusts are formed of flakes or dust composed of epithelial
debris.
Most lesions occur on the back and sides.
Spontaneous recovery does not occur in adult pigs.
1466
 Ringworm Cont’d---
3.In sheep and goats
In sheep the lesions occur on the head rarely in the fleeced
areas.
However, they usually disappear in 4 - 5 weeks.
But the disease may persist in the flock for some months.
The lesions are discrete, round, almost bald patches covered
with a grayish crust.
Similar lesions occur in goats but they are distributed
generally over all parts of the body.
The exception to ringworm of sheep is a new ringworm
associated with an unidentified Trichophyton spp. in some
countries of the world.
Lesions occur extensively in fleeced areas and are
characterized by shedding of the wool staple and exudation
from the skin surface.
1467
 Ringworm Cont’d---
Necropsy findings
There is no as such pathological changes in internal organs except
lesions on the skin.
Diagnosis
Diagnosis of ringworm in animals is done based on
 Epidemiology of the disease
Clinical signs of the disease(the appearance of characteristic
lesions)
Result of laboratory findings
Diagnostic confirmation is done in laboratory by demonstration of
fungal elements in a scraping or biopsy which include
 Demonstration of spores and mycelia in skin scrapings and in
culture.
Isolation of culture of fungi on/in artificial media.
Specimens for fungi isolation and identification in dermatophytoses
is skin scrap that are taken peripherally from animals those were
not treated. 1468
 Ringworm Cont’d---
Skin scrapings should be made after defatting the skin with ether
or alcohol and gently warming the scraping in a 10 - 20% solution
of either potassium or sodium hydroxide(KOH or NaOH).
Specimens to be sent for laboratory examination should be packed
in envelopes.
Because airtight jars and cans favor the growth of non-pathogenic
fungi.
For identification of species of dermatophytes we must culture the
specimen on/ in artificial media.
The species of the fungi is identified by
 Growth rate of the fungi on artificial media
 Colour and morphology of the colony
 Character of mycelia
 Morphology of micro and macroconidia, arthrospores and
chlamydospores.
For example morphology of spores under microscope is the
diagnostic feature to differentiate the species of fungi. 1469
 Ringworm Cont’d---
If the spore appears as round or polyhedral, highly refractive bodies
in chains under microscope it is Trichophyton spp.
But if the spore appears as mosaics it is Microsporum spp.
Fungal hyphae in tissues can be identified, even down to the genus
by the use of immunofluorescent staining.
The technique was devised for use on necropsy material but should
have application for biopsy material and scrapings.
Differential diagnosis
The differential diagnosis list of ringworm which may be confused
with diseases with similar clinical profiles varies and depends on
species of animals.
In cattle, you must consider the following diseases as differential
diagnosis of ringworm.
Mycotic dermatitis
 Inherited parakeratosis
Sarcoptic mange
Psoroptic mange 1470
 Ringworm Cont’d---
Mycotic dermatitis is differ from ringworm in that it has tenacious
scabs which cover a raw area of skin.
Inherited parakeratosis is differ from ringworm in that it is
characterized by tenacious thick crusts which respond quickly and
completely to dietary supplementation with zinc.
Sarcoptic mange is differ from ringworm in that mites can be
demonstrated in scrapings and there is intense pruritus and a quick
response to standard insecticides
Psoroptic mange is differ from ringworm in that it is identifiable
by the presence of mites in scrapings, pruritus occurrence in housed
cattle and the location of the lesions over the hindquarter.
In pigs the following diseases must be taken as differential
diagnosis.
 Pityriasis rosea, in which no mites can be demonstrated and the
disease is limited to a particular age group
Exudative epidermitis has extensive lesions with a characteristic
greasy covering 1471
 Ringworm Cont’d---
Tyrogliphosis is self-limiting, associated with a new source of
grain, and characterized by pruritus
Sarcoptic mange, identifiable by the mites in scrapings, the
intense pruritus and the prompt response to treatment with
insecticide
In sheep and goats ringworm must be differentiated at
Dermatitis of non – infectious etiology
Sheep keds
Mangemitis etc.
Treatment
Treatment is widely practiced and recommended.
Because it greatly reduces the contamination of the environment
by infected animals.
Depending on the size of the lesion treatment of ringworm can be
1. Local treatment
2. Systemic treatment
1472
 Ringworm Cont’d---
1. Local treatment
It can be
 Local application
 Local spray or washes
Local application is topical application of medicaments.
Local application is widely used when the lesions of the skin are
localized.
Topical treatment is probably of greater value in the early stages of
an outbreak when the lesions are small and few in number.
The crusts should be removed by scraping or brushing with a soft
wire brush and burned.
The medicament should be brushed or rubbed in vigorously.
The suitable medicaments that are used for topical applications
include
A weak solution of iodine
Whitfield's ointment
10 % ammoniated mercury ointment 1473
 Ringworm Cont’d---
Solutions of quaternary ammonium compounds (1:200-1 :1000)
 Propionic and undecylenic acid ointments
Solutions of 0.25% hexadecamethylene
Bisisoquinolinium chloride
Borotannic complex
 1-5 % ointment the antibiotics (natamycin and nanomycin A)
Povidone-iodine
Thiabendazole
Ointments containing one of the azole compounds such as
imidazole or miconazole.
Local spray or washes is used when infection in a group is
widespread.
Washes or sprays which can be applied over the entire body surface
of all animals are used when ringworm infection is wide spread.
But the efficacy of the medicament is less than that of ointments
and daily application for at least 5 days is required.
However, sprays have a big advantage if prophylactic treatment1474
 Ringworm Cont’d---
of all in-contact animals is recommended.
The most common preparates that are used for spraying are
5% lime sulfur (20 % w/v polysulfides diluted 1:20)
Captan
3% N-trichloromethylthio- tetrahydrophthalimide
Iodofors
 0.5% sodium hypochlorite
Natamycin (100 ppm)
2. Systemic treatment
Systemic treatments recommended for use in farm animals include
the intravenous injection of 10% sodium iodide at the dose of 1
g/14 kg body weight repeated on several occasions.
If the cost of the drug is not problem oral administration of
griseofulvin has high efficacy.
1475
 Ringworm Cont’d---
Control and preventation measures
Control and preventation measures of ringworm can be achieved
by
Hygiene
Vaccination
Nutrition
Failure to control an outbreak of ringworm is usually due to the
widespread contamination of the environment before treatment is
attempted.
The following practices help to control ringworm
Isolation and treatment of infected animals
 The provision of separate grooming tools, horse blankets and
feeding utensils
Disinfection of these items after use on affected animals
The following chemicals are used to clean and to disinfect the
fungi
 Commercial detergent or a strong solution (2.5-5 %) of phenolic1476
 Ringworm Cont’d---
5% lime sulfur
5% formalin
3% Captan
 5% sodium hypochlorite
Good results are also claimed for the disinfection of buildings with
a spray containing
2.0% formaldehyde and 1.0% caustic soda.
Vaccination to prevent ringworm has been practiced in many
countries of Europe.
The vaccines include those containing highly immunogenic, non
virulent strains, or attenuated strains of fungi, or those killed
vaccines containing specific fractions of mycelia.
Vaccination of all animals in the group is recommended and
isolation and treatment of infected animals and disinfection of
premises.
The vaccine is almost totally without side effects except for very
rare deaths due to anaphylaxis. 1477
 Ringworm Cont’d---
Nutrition also play essential role in controlling ringworm.
Ringworm occurs in well nourished as well as poorly fed animals.
But there seem to be a tendency for the latter to become infected
more readily and to develop more extensive lesions.
Supplementation of the diet, particularly with vitamin A to young
housed animals, should be encouraged as a preventive measure of
fungal infection.

1478
1479
7.1. Parturient paresis(milk fever)
7.2. Ketosis
7.3. Copper deficiency
7.4. Cobalt deficiency
7.5. Vitamin A deficiency
7.6. Hypomagnesaemia tetani
7.7 . Selenium/vitamin E deficiency

1480
 Parturient paresis(milk fever) is a disease of cattle, sheep and goats
occurring around the time of parturition.
It is caused by hypocalcemia and characterized by
Weakness
Recumbency
Ultimately shock and death
Etiology
It is caused by hypocalcemia just before or after parturition.
So that a depression of the levels of ionized calcium in tissue fluids
is the basic biochemical defect in milk fever.
A transient period of subclinical hypocalcemia is characterized by
less than 1.9 mmol of calcium ion /liter of blood plasma during
onset of lactation .
It is caused by an imbalance between calcium output in the
colostrum and influx of calcium to the extracellular pool from
intestine and bone.
1481
 Milk fever Cont’d---
That is why the onset of lactation results in a sudden large demand
on the calcium homeostasis.
This is about nine times as much calcium as that present in the
entire plasma calcium pool of the cow is required during the onset
of lactation.
Calcium lost from the plasma pool for colostrum production must
be replaced by increasing intestinal absorption and bone resorption.
During the dry period, calcium requirements are minimal at about
10-12 gm/day.
However at parturition, the cow must mobilize about 30 gm/ day
or more of calcium into the calcium pool per day.
You know hypocalcemia may occur because of inappropriate
function of
Endocrine system like parathyroid and thyroid gland which
produces parathyroid and calcitonin hormones respectively.
Deficiency of calcipherol(vitamin D)
Failure of the absorvation of Ca ion through small intestine due 1482
to
 Milk fever Cont’d---
different reasons.
But hypocalcemia which occurs in spite of apparently adequate
function of the parathyroid gland; absence of failure in its
absorvation and with appropriate amount of vitamin D most cows
adapt within 48 hours after calving by increases in plasma
concentrations of parathyroid hormone.
These cows during the onset of the hypocalcemia, they mobilize
calcium by increasing intestinal absorption and bone resorption.
But about 5 - 20% of adult cows are unable to maintain plasma
calcium by the above process due to different reasons and
consequently develop severe hypocalcemia.
Such cows have a total plasma calcium ion which is reduced 1.00
-1.40 m mol/ liter of plasma.
As the result clinical milk fever called parturient paresis which
requires immediate treatment develops.
Epidemiology
In epidemiology of milk fever, we must consider 1483
 Milk fever Cont’d---
Species of animals affected
Age of animals at which susceptibility is increased
Breed
Individual animals
Time of occurrence
Stressors those predispose animals to milk fever
Milk fever affects cattle, sheep and goats.
The disease occurs most commonly in high-producing adult
lactating dairy cattle.
Lactating beef cows are affected but less commonly.
There are differences in susceptibility between the breeds but the
differences are small.
Zebu breed is less susceptible to milk fever than cross breeds and
exotics.
From exotics Jersey breeds are the most susceptible breeds.
The heritability of susceptibility to milk fever and hypocalcemia has
been assessed and it is insignificant. 1484
 Milk fever Cont’d---
Several studies have reported that the incidence of milk fever is
positively associated with the level of milk production.
This means high producing dairy cows are more susceptible to milk
fever.
In cattle, milk fever occurs at three main stages in the lactation
cycle.
These are
In last few days of pregnancy
Rare cases occur several weeks before calving
Some cases will occur a few hours before parturition or at the time
of parturition
Milk fever cases occur within the first 48 hours after calving and the
danger period extends upto about the 10th days of postpartum
period.
There are many stress factors that predispose the pregnant cows to
milk fever.
Some of the examples are 1485
 Milk fever Cont’d---
1. Starvation for 48 hours before parturition
It causes severe depression of serum calcium level and this may be
importance in the production of hypocalcemic paresis in dairy cows
at time of parturition.
2. In winter months when pregnant beef cattle are fed on poor-
quality roughage
Feeding of poor quality roughage feeds to pregnant beef cows
results in hypocalcemic paresis(milk fever)
Among the groups of such cows the less aggressive ones may suffer
selective malnutrition.
3. Milk fever may occur in beef cows affected with diarrhea of
undetermined etiology.
But you must understand that hypocalcemic syndromes in
ruminants are also observed at times other than related to
parturition.
Some of the examples are
Overeating of fermentable carbohydrate due to the formation of1486
 Milk fever Cont’d---
lactate salts.
Intera venous administration of certain aminoglycosides, especially
neomycin, elihydrostreptomycin and gentamycin.
They may cause a reduction in the degree of ionization of serum
calcium and a syndrome similar to milk fever.
NB. So that a veterinarians must be careful not to administer the
above drugs during parturient period of cows.
In sheep, the disease commonly occurs in outbreaks in groups of
ewes exposed to
Forced exercise
Long-distance transport
Sudden deprivation of food
Grazing on oxalate-containing plants or green cereal crops
At the end of a drought when the pasture growth is lush and very
low in calcium content.
These circumstances commonly precipitate outbreaks of
1487
hypocalcemic paresis in sheep.
 Milk fever Cont’d---
Mature ewes are the most susceptible, particularly in the period
from 6 weeks before to 10 weeks after lambing.
The disease is manifested by paresis.
But in the rest of the flock may show poor growth, lameness and
bone fragility.
Milking goats become affected mostly during the 4-6-year age
group.
Cases occur before and after kidding, some later than 3 weeks after
parturition.
Clinical syndromes are identical to those in cows, including the two
stages of ataxia and recumbency.
Generally milk fever has sporadic occurrence.
But in intensive farms the incidence of milk fever may rarely reach
25 - 30% in high producing dairy cows.
From the total 75% to 85% of uncomplicated cases respond to
calcium therapy very well.
The remaining 15 - 25% respond bad to calcium therapy, this 1488
 Milk fever Cont’d---
because
 Milk fever is complicated by other conditions or
Incorrectly diagnosed by veterinarians
Risk factors
The risk factors for occurrence of milk fever are
1. Animal risk factors
2. Dietary( Nutrition) risk factor
1. Animal risk factors
Serum calcium level declines in all adult cows at calving due to the
onset of lactation.
Serum calcium levels decline to lower levels in some cows than in
others .
This shows some cows are more susceptible to parturient paresis
than others.
Milk fever susceptibility increases with age and first calving heifers
are less susceptible to parturient paresis.
Because they are able to adapt rapidly to the high demands of 1489
 Milk fever Cont’d---
calcium for lactation.
However, with increasing age, this adaptation process is decreased.
This results in moderate-to-severe hypocalcemia in most adult cows.
The adaptation mechanism is directly related to the efficiency of
intestinal absorption of calcium, which decreases with increasing
age.
Three factors affect calcium homeostasis
1.Excessive loss of calcium in the colostrum beyond the capacity of
absorption from the intestines and mobilization from the bones to
replace
Variations in susceptibility between cows could be due to variations
in the concentration of calcium in the milk and the volume of milk
secreted.
2.Impairment of absorption of calcium from the intestine at
parturition.
3.Mobilization of calcium from storage in the skeleton may not be
sufficiently rapid to maintain normal serum levels. 1490
 Milk fever Cont’d---
The calcium mobilization rate and the immediately available
calcium reserves are sufficiently reduced in cows in later pregnancy
to render them incapable of withstanding the expected loss of
calcium in the milk.
In older cows, bone resorption makes only a minor contribution to
the total rate of calcium mobilization at parturition .
Therefore of minor importance for the prevention of periparturient
hypocalcemia.
Osteoblasts are the only type of bone cell to express the 1,25-
(OH)2D receptor protein.
The decrease in the numbers of osteoblasts with increasing age
could delay the ability of bone to contribute calcium to the plasma
calcium pool.
Body condition score (BCS) is also one of animal risk factor that
affect the occurrence of milk fever.
A high BCS increases the risk of milk fever.
1491
 Milk fever Cont’d---
2. Dietary( Nutrition) risk factor
Several dietary factors of the pregnant cow during the peripartum
period (last 4 weeks) can influence the incidence of milk fever in
cattle.
Feeding more than 100 gm of calcium daily during the dry period
is associated with an increased incidence of milk fever.
A 500 kg cow requires only about 31 gm of calcium to meet daily
maintenance and fetal demands in late gestation.
When a cow is fed a high calcium diet (>100 gm Ca/day), its daily
requirement for calcium can be met almost entirely by passive
absorption of dietary calcium.
Th active transport of calcium from the diet and bone calcium
resorption mechanisms are homeostatically depressed and become
quiescent.
As a consequence, at calving, the cow is unable to use bone
calcium stores or intestinal calcium absorption mechanisms and is
susceptible to severe hypocalcemia until these mechanisms can1492
be
 Milk fever Cont’d---
activated which may take several days.
Feeding peripartum diets containing a low concentration of calcium
prevents milk fever by activating calcium transport mechanisms in
the intestine and bone prior to parturition.
Thus allowing the animal to adapt more rapidly to the lactation
drain of calcium.
Feeding diets high in calcium just before parturition may also lower
the incidence of milk fever by increasing th absorption of calcium.
In sheep, hypocalcemia may occur in pregnant ewes fed a calcium-
deficient diet over a prolonged period.
Another nutritional risk factor is dietary phosphorus in the feed.
Peripartum diets which is high in phosphorus (>80 gm P/day) also
increases the incidence of milk fever and the severity of
hypocalcemia.
High dietary levels of phosphorus increases the serum level of
phosphorus which is inhibitory to the renal enzymes that catalyze
production of 1,25- (OH)2D decreased reduce the intestinal calcium
1493
 Milk fever Cont’d---
absorption mechanisms peripartum.
Economic importance
The economic loss due to milk fever comprises
 The need of repeating treatment in some cases because of
relapsing milk fever.
Economic loss due to downer cow complications.
The downer cow syndrome is associated with milk fever cases.
It is a syndrome that fails the cow to respond within a few hours
and remain recumbent for several hours or several days before
subsequently standing, or die, or are euthanized.
It represents an important cause of economic loss because of
Treatment costs
 Long-term care costs
Loss of milk production
 Loss of the value of the animal.
Acute mastitis due to environmental pathogens is a common
complication of downer cow complications
1494
 Milk fever Cont’d---
Milk fever predispose the cow to dystocia and reproductive disease.
Hypocalcemia at the time of parturition can result in uterine inertia
which may cause dystocia and uterine prolapse.
Parturient paresis predispose the cow to retained placenta.
Several studies have found an increased risk of retained placenta
following milk fever.
Milk fever predispose the cow to metritis.
A few studies have found an indirect relationship between milk
fever and subsequent metritis.
Milk fever causes loss of milk production due to mastitis and
prolonged recumbency.
Milk fever also the main causes of displacement of abomasum,
body weight.
Parturient paresis predispose the cow to ketosis.
There may be an increased probability of culling cows which have
had milk fever.
Because of the complications or direct or indirect consequences
associated with the disease
1495
 Milk fever Cont’d---
Pathogenesis
There is no similar idea in pathogenesis of milk fever.
However, many researchers believe that suddenly disturbance in
mineral and sugar regulation causes in disturbance neuro – endocrine
system regulation.
This disturbance cause in
Hypocalcemia
Hyper magnaemia
Hypo glycemia
In correct ratio of calcium and magnesium in blood results in
Disturbance in Ca and Mg metabolism
Depression of CNS
All this results in disturbance the function of parathyroid and
hypophises which ends with coma.
We know hypocalcemia occurs because of
The increased amount of Ca ion together with colostrum
Inhibition of the function parathyroid gland 1496
 Milk fever Cont’d---
While hypoglycemia is due to
High production of insulin which causes less amount of glucose in
blood.
The disturbance of these macro elements like Ca, P and Mg; and
sugar cause depression of the function of hypothalamus –
hypophises and adrenaline gland relation ship.
 This ends with stress and disease which is commonly called
parturient paresis(milk fever).
Clinical findings
Parturient paresis usually occurs within 12 - 72 hrs of parturition.
Rarely before or at parturition time.
There are 3 discernible stages of parturient paresis.
First stage
Second stage
Third stage
During the first stage of parturient paresis, the main clinical
signs are 1497
 Milk fever Cont’d---
Animals are ambulatory but show signs of hypersensitivity and
excitability
Cows may be mildly ataxic, have fine tremors over the flanks and
triceps, and display ear twitching and head bobbing.
Cows may appear restless, shuffling their rear feet and bellowing.
If calcium therapy is not instituted, cows will likely progress to the
second, more severe stage.
During the second stage of parturient paresis, the main clinical
signs are
The cow unable to stand but can maintain sternal recumbency
Cows are obtunded, anorectic and have a dry muzzle
Subnormal body temperature and cold extremities
Auscultation reveals tachycardia and decreased intensity of heart
sounds
Peripheral pulses are weak
Paralysis smooth muscle leads to GI stasis which can manifest as
bloat, failure to defecate and loss of anal sphincter tone. 1498
 Milk fever Cont’d---
 An inability to urinate may manifest as a distended bladder on
rectal examination.
Cows often tuck their heads into their flanks, or if the head is
extended an S-shaped curve to the neck may be noted.
During the third stage of parturient paresis, the main clinical
signs are
Cows loses consciousness progressively to the point of coma.
They are unable to maintain sternal recumbency
They have complete muscle flaccidity
The cows are unresponsive to stimuli and can suffer severe bloat
As cardiac output worsens, heart rate can approach 120
pulse/minute peripheral pulses may be undetectable.
If untreated, cows in stage 3 may survive only a few hours
Parturient paresis in ewes and goats is characterized by similar
clinical signs like cow.
However parturient paresis in pigs occurs mostly after 2 – 5 days
of parturition and has the following clinical features. 1499
 Milk fever Cont’d---
In pigs, as in cattle, signs develop within a few hours of farrowing.
Generally, the main clinical signs are
 Restlessness
Anorexia
 Inability to rise followed by lateral recumbency
 Coma
But for easy under standing of milk fever in pigs can be
conditionally to be classified as easy and complicated form of milk
fever.
In easy form of parturient paresis:-
The pig stands with difficulty and loss its coordination.
In complicated form of parturient paresis: -
 The pig tries to stand but it may have sitting dog posture and unable
to move.
Body temperature decreases and may reach 37 0C
Peristalsis is reduced and defect with difficulty as the result the
faces become dry. 1500
 Milk fever Cont’d---
Necropsy findings
No specific lesions.
But ischemic muscle necrosis of large muscles of pelvic limbs may
occur because of prolonged recumbency.
Diagnosis of parturient paresis
A diagnosis of milk fever is based on the occurrence of paresis and
depression of consciousness in animals following parturition.
Diagnosis is supported by
A favorable response to treatment with parenteral injections of
calcium solutions
 Biochemical examination of the blood
So that diagnostic confirmation of parturient paresis is
hypocalcemia in serum and response to treatment with calcium
borogluconate.
Differential diagnosis
In the immediate postpartum period, there are several diseases
which cause recumbency in cows. 1501
 Milk fever Cont’d---
So that diseases of cows, sheep and goats those must be
differentiated from parturient paresis are
1. Cattle
From metabolic and nutritional disease like
Hypophosphatemia
Hypomagnesaemia
Downer cow syndrome
 Fat cow syndrome
Carbohydrate engorgement.
Toxemias
Peracute coliform mastitis
Aspiration pneumonia
 Acute d iffuse peritonitis.
Injuries to pelvis and pelvic limbs
 Maternal obstetrical paralysis
 Dislocation of coxofemoral joint
Disease caused by prions
1502
Bovine spongiform encephalopathy
 Milk fever Cont’d---
2.Sheep and goats
Pregnancy toxemia
Enterotoxemia.

1503
 Milk fever Cont’d---
Differential diagnosis of common causes of recumbency in
parturient adult cattle
Disease Clinical signs Clinical Response to treatment
pathology
Milk Early excitement and Hypocalcemia, Rapid response
fever(parturie tetany, coma, High serum characteristic(Muscle tremor
nt paresis) hypothermia, flaccidity, Magnesium sweating on muscle
pupil dilation, weak Low inorganic ,defecation ,urination, pulse
heart sounds . phosphate in amplitude and heart sound
No rumen movement serum intensity improve first) after
IV injection soluble Ca salts

Downers cow Moderately bright, Variable. May be Variable response to

syndrome active, eating. Temp. low inorganic calcium, phosphorus and
following slightly raised, HR 80-1 phosphate, potassium salt.
milk fever 00/ m. Unable to stand potassium, or Fluid therapy and provision
but tries -'creepers'. glucose, of deep bedding and hourly
When dull and Ketonuria, rolling from side to side are
depressed, are non-alert usually necessary.
downers'. Long course 1-
2 weeks 1504
 Milk fever Cont’d---
Table Cont’d
CH2O Complete cessation of Haemoconcentration Ruminotomy or rumen
engorgement ruminal activity. with severe acidosis, lavages
Fluid splashing sounds in PH of rumen juice may be necessary.
rumen below 5. Alkalinizing
Severe dehydration No living protozoa agents must be
circulatory failure. in rumen demonstrated
Apparent blindness then
recumbency and too weak
to rise soft odor ferrous
feces
Hypomagnes Excitement, Low serum Even after IV injection
aemia hypersensitivity magnesium response in a severe
(lactation, muscle tremor, tetany. case may take 30 min,
grass tetany) Recumbent with tetanic much slower than
convulsions, loud heart response to calcium in
sounds, rapid rate, milk fever
Subacute cases remain
standing

1505
 Milk fever Cont’d---
Table Cont’d
Severe Recumbency, depression to Profound Require supportive response
toxemia coma, sleepy, dry nose, leukopenia for toxemia and shock.
(acute diffuse hypothermia, gut stasis, HR Response is poor and
peritonitis, over 100/min, may be temporary.
coliform grunting. Prognosis very bad.
mastitis) Examine mammary gland. May die if treated IV with
Examine abdomen for calcium or magnesium salt.
abdominal disease

Fat cow Excessive body condition, Evidence of Will recover if begin to eat.
syndrome anorexia, apathy, hepatic Treat with fluids, glucose,
depression, disease. insulin.
recumbency and looks like Provide good quality
milk fever, scant soft feces, palatable roughage
ketonuria

1506
 Milk fever Cont’d---
Bovine spongi form Most die or are Laboratory Nil
encephalopathy euthanatized Insidious examination of brain
onset, clinical course for presence of prions
several weeks, changes
in behavior
hyperesthesia, ataxia,
loss of weight, kick
during milking,
knuckling, progressive
leading to recumbency

Treatment
Parturient paresis can be treated by administration of
15 – 20 ml of 20% caffeine SC route to stimulate heart function
20 0 – 250 ml of 40% glucose IV route
100 – 150 ml 10 % Cacl2 IV route strictly
 100 – 150 ml of 10% calcium gluconate or calcium borogluconate
IV route.
1507
 Milk fever Cont’d---
If the animal recovers after treatment
Response to environmental stimuli increases.
Body temperature comes to norm
The cow starts to move tongue.
Starts to defecate and urinate
 Rumination begins
Parturient paresis in ewes, goats and pigs can be treated similar to
cows.
If parturient paresis is not treated on time, it may predispose the
cow to
Dystocia
Uterine prolapse
Retained fetal membranes
 Metritis
Abomasal displacement
 Mastitis
1508
 Milk fever Cont’d---
Control and preventation measures
Various methods are available for the control of milk fever in
ruminants, especially in dairy cows.
They include
Dietary management during the transition period before and after
calving
A administration calcium gels orally at the time of parturition
 Administration of vitamin D and its metabolites and analogs
immediately before parturition to enhance the mobilization of
calcium
Options for reducing the risk of hypocalcemia in pasture-fed dairy
cows are
1. Low calcium intake peripartum
2. Magnesium supplementation
3. Calcium supplementation
1. Low calcium intake peripartum
A low calcium diet in the peripartum period will activate calcium1509
 Milk fever Cont’d---
homeostasis at calving.
An alternative is to feed calcium binding agents for 1-3 weeks
peripartum.
2. Magnesium supplementation
Hypomagnesaemia influences calcium homeostasis, potentially
predisposing the cow to milk fever at calving.
Diets high in potassium can reduce the concentration of plasma
magnesium and may be a mechanism linking high dietary
potassium to hypocalcemia.
Recommended concentrations for dietary magnesium levels are
within the range of 0.2 - 0.4% of total DM intake.
Plasma levels are readily elevated when magnesium is added to
the diet.
3. Calcium supplementation
Oral administration of gels of calcium chloride or calcium
propionate immediately after calving is effective in preventing
milk fever. 1510
 Milk fever Cont’d---
Feeding cows an excess of calcium in the form of calcium
carbonate during the first few weeks of lactation is beneficial.
Increasing dietary concentration of calcium from 0.68% to 1.02%
DM within 8 h of calving has an immediate effect on plasma
calcium.
Generally, the following management practices are suggested to
prevent parturient paresis
Avoid over fattening by either reducing the energy concentration of
the ration or restricting the intake during the peripartum period
A void stresses at the time of parturition
 Provide a clean well-bedded box stall with conditions conducive to
cow comfort and allow the animal to exercise
 Make frequent observations of cows prone to milk fever from 48 h
before to 48 h after parturition for evidence of milk fever and
immediate treatment will reduce the incidence of the downer cow
syndrome associated with milk fever.
1511
 Milk fever Cont’d---
 At calving the cow should receive an oral dose of a calcium salt in
a gel, as set out later, followed by a diet with a high calcium
content (over 1 % of dry matter) .
 The critical day is the day of calving and a sharp increase in
calcium intake on this day can significantly reduce the occurrence
of milk fever.
 If hypomagnesaemia is a likely concomitant, the diet should be
supplemented with 60 gm magnesium oxide daily.
7.2. Ketosis(Acetonemia)
Ketosis is a common disease of adult cattle; occurs in dairy cows
in early and rarely in late lactations.
It is consistently characterized by partial anorexia and depression.
In addition to inappetance, signs of nervous dysfunction are
occasionally seen, including
 Pica and abnormal licking
 Incoordination and abnormal gait
1512
 Ketosis Cont’d---
Bellowing and aggression
Etiology
A multifactorial disorder of energy metabolism.
It is a negative energy results in hypoglycemia and Ketonaemia.
As the result there is high accumulation in blood of acetoacetate, β-
hydroxybutyrate and their decarboxylation products acetone and
isopropanol.
Depending on the geneses of ketosis, ketosis can be classified as
1. Primary ketosis (production ketosis)
2. Secondary ketosis
3. Alimentary ketosis
4. Starvation ketosis
5. Ketosis due to specific nutritional deficiency
1. Primary ketosis (production ketosis)
This is the ketosis of most herds, the so called estate acetonemia.
1513
It occurs in cows in good to excessive body condition that have high
 Ketosis Cont’d---
lactation potential and are being fed good-quality rations but that
are in a negative energy balance.
A proportion of cases appear as clinical ketosis.
But a much greater proportion occur as cases of subclinical ketosis
in which there are increased levels of circulating ketone bodies but
no overt clinical signs.
2. Secondary ketosis
This occurs where other disease results in a decreased food intake.
The cause of the reduction in food intake commonly in dairy cows
are the result of
Abomasal displacement
Traumatic reticulitis
Metritis
Mastitis and other diseases' common to the post parturient period
3. Alimentary ketosis
This form is due to excessive amounts of butyrate in silage.
It is also possibly also due to decreased food intake resulting from
1514
 Ketosis Cont’d---
poor palatability of high butyrate silage.
Silage made from succulent material may be more highly
ketogenic than other types of silage Because of its higher content
of preformed butyric acid.
Spoiled silage is also a cause and toxic biogenic amines in silage,
such as putrescine, may also
Alimentary ketosis is commonly subclinical.
But it may predispose to the development of production or
primary ketosis.
4. Starvation ketosis
This occurs in cattle that are in poor body condition and that are
fed poor-quality feedstuffs.
There is a deficiency of propionate and protein from the diet and a
limited capacity of gluconeogenesis from body reserves.
5. Ketosis due to specific nutritional deficiency
Specific dietary deficiencies of cobalt and possibly phosphorus
may also lead to a high incidence of ketosis. 1515
 Ketosis Cont’d---
This may be due in part to a reduction in the intake of total
digestible nutrients (TDN).
But in cobalt deficiency, the essential defect is a failure to
metabolize propionic acid into the tricarboxylic acid (TCA) cycle.
The problem is restricted to the cobalt deficient areas of the world.
But the occurrence of cobalt deficiency in high-producing dairy
cows in non-deficient areas has been described.
Epidemiology of ketosis
All dairy cows in early lactation during the first 6 weeks are at
risk of ketosis.
Although it appears to be less common in primiparous animals,
ketosis is seen in all parities.
It does not appear to have a genetic predisposition other than being
associated with dairy breeds.
Cows with excessive adipose stores (body condition score ≥3.75
out of 5) at calving are at a greater risk of ketosis than those with
lower body condition scores. 1516
 Ketosis Cont’d---
Risk factors
The common risk factors for occurrence of ketosis are
1.Animal risk factors
2. Managemental and environmental risk factors
1. Animal risk factors
These are
Stage of lactation
Age of animals
Body condition score (BCS) of animals
The disease occurs in the immediate post parturient period.
However, 90% of cases occurring in the first 60 days of lactation.
Generally, regardless of specific etiology, ketosis occurs most
commonly
During the first month of lactation
 Less commonly in the second month
 Only occasionally in late pregnancy.
1517
 Ketosis Cont’d---
Cows of any age may be affected.
But the disease increases from a low prevalence at the first calving
to a peak at the fourth and reduces after the fourth calving.
Increased body condition loss and a higher risk for ketosis .
In another study, cows with good BCS at parturition and that lost
her BCS in the first 2 months of lactation developed subclinical
ketosis
Body condition loss during the dry period also causes ketosis
following lactation.
2. Managemental and environmental risk factors
Herd differences in prevalence of ketosis are very evident in
clinical practice.
Big herd cows are more prone to ketosis.
There is no clear association with season.
Higher risk of ketosis is generally observed in cattle during the
winter housing period.
There is a greater risk for the development of ketosis in cows that
1518
 Ketosis Cont’d---
have
 An extended long dry period
 Develop milk fever,
Retained placenta,
Lameness or hypomagnesaemia.
Cows with twins are also at risk for ketosis in the terminal stages of
pregnancy.
Economic significance
Clinical and subclinical ketosis are major causes of loss to the dairy
farmer.
In rare instances the disease is irreversible and the affected animal
dies.
But the main economic loss while the disease is present is due to
The loss of production
The occurrence of periparturient disease
Both clinical and subclinical ketosis are accompanied by
Decreased milk yields 1519
 Ketosis Cont’d---
Lower milk protein and milk lactose
Increased risk for delayed estrus
Lower first service conception rates
Increased inter-calving intervals
Increased risk of cystic ovarian disease
Metritis and mastitis and increased involuntary culling
Pathogenesis
The pathogenesis of bovine ketosis is incompletely understood.
But it requires the combination of intense adipose mobilization and
a high glucose demand.
So in bovine ketosis, the principal metabolic disturbances observed
are hypoglycemia and ketonaemia.
Both of these conditions are present in early lactation and at the
period of peak lactation.
It is a time of negative energy balance which leads to adipose
mobilization and milk synthesis creates a high glucose demand.
1520
 Ketosis Cont’d---
Adipose mobilization is accompanied by high blood serum
concentrations of Non Esterified Fatty Acids (NEFAs).
During these periods of intense gluconeogenesis, a large portion of
serum NEFAs are directed to ketone body synthesis in the liver.
Thus, the clinicopathological characterization of ketosis includes
high serum concentrations of NEFAs and ketone bodies and low
concentrations of glucose.
The serum ketone bodies are
Acetone
Acetoacetate
β-hydroxybutyrate (BHB)
In many cases, the severity of the clinical syndrome is proportional
to the degree of hypoglycemia.
It is suggested that hypoglycemia is as the predominant factor in
bovine ketosis.
In most field cases the severity of the clinical syndrome is also
1521
 Ketosis Cont’d---
roughly proportional to the degree of ketonaemia.
This is an understandable relationship as ketone bodies are
produced in larger quantities as the deficiency of glucose increases.
However, the ketone bodies may exert an additional influence on
the signs observed.
Acetoacetic acid is known to be toxic and probably contributes to
the terminal coma.
The nervous signs which occur in some cases of bovine ketosis are
caused by
The production of isopropyl alcohol, a breakdown product of
acetoacetic acid in the rumen
 The requirement of nervous tissue for glucose to maintain normal
function may also be a factor in these cases.
Spontaneous ketosis in cattle is usually readily reversible by
treatment.
But incomplete or temporary response is usually due to the
existence of a primary disease with ketosis present only as a 1522
 Ketosis Cont’d---
a secondary development,
Changes in ruminal flora after a long period of anorexia may also
cause continued impairment of digestion.
Immunosuppression has been demonstrated with energy deficiency
and ketosis.
The higher susceptibility of ketotic postpartum cows to local and
systemic infections may be related to impairment of the respiratory
burst of neutrophils which occurs with elevated levels of β -
hydroxybutyrate (BHBA).
Depending on the time of occurrence, ketosis in cattle can be
classified as
Type I ketosis
Type II ketosis
Ketosis cases occurring closer to peak milk production.
That means ketosis which usually occurs at 4–6 weeks postpartum
period.
It may be more closely associated with under fed cattle 1523
 Ketosis Cont’d---
experiencing a metabolic shortage of gluconeogenic precursors than
with excessive fat mobilization.
Ketosis at this time is sometimes described as type I ketosis.
Ketosis cases occurring in the immediate postpartum period is
slightly different than that of cases occurring closer to the time of
peak milk production.
Ketosis in the immediate postpartum period is sometimes
described as type II ketosis.
Such cases of ketosis in very early lactation are usually associated
with fatty liver.
Both fatty liver and ketosis are probably part of a spectrum of
conditions associated with intense fat mobilization in cattle.
Clinical findings
Ketosis can be
 1. Clinical
 2. Subclinical
1524
 Ketosis Cont’d---
1. Clinical ketosis
Two major clinical forms of bovine ketosis are described.
These are
1.The wasting ketosis
2. The nervous ketosis
1.The wasting ketosis
But these are the two extremes of a range of syndromes in which
wasting and nervous signs are present in varying degrees of
prominence.
Body weight is lost rapidly, usually at a greater rate than one would
expect from the decrease in appetite.
Farmers usually describe affected cows as having a ‘woody’
appearance.
The temperature, the pulse and respiratory rates are normal.
But the ruminal movements may be decreased in amplitude and
number
A characteristic odor of ketones is detectable on the breath and 1525
 Ketosis Cont’d---
often in the milk.
2. The nervous form of ketosis
Signs are usually bizarre and begin quite suddenly.
The syndrome is suggestive of delirium rather than of frenzy and
the characteristic signs include:
Walking in circles
Straddling or crossing of the legs
Head pushing or leaning into the stanchion
Apparent blindness
Aimless movements and wandering
Vigorous licking of the skin and inanimate objects
Depraved appetite
Chewing movements with salivation
Hyperesthesia may be evident
The animal bellowing on being pinched or stroked.
Moderate tremor and tetany may be present
 There is usually an in coordinate gait. 1526
 Ketosis Cont’d---
The nervous signs usually occur in short episodes which last for 1
or 2 hours.
It may recur at intervals of about 8 -12 hours.
Affected cows may injure themselves during the nervous episodes.
2. Subclinically ketosis
It ketosis in many cows in early pregnancy with out showing
clinical signs.
However these cows are
Negative in energy balance
They have ketonuria
They have diminished productivity including depression of milk by
1. 9%.
 Reduction in fertility.
Infertility may appear as an ovarian abnormality and is
characterized by
 Delayed onset of estrus
Endometritis (resulting in an increase in calving to conception 1527
 Ketosis Cont’d---
interval and reduced conception rate at first insemination).
Necropsy findings
The disease is not usually fatal in cattle.
But fatty degeneration of the liver and secondary changes in the
anterior pituitary gland and adrenal cortex may be present.
Diagnosis
Diagnose of bovine ketosis is done based on history, clinical signs
and laboratory findings.
The clinical picture is usually too indefinite especially in cattle, to
enable a diagnosis to be made on clinical grounds.
General consideration is necessary to diagnose ketosis like history
with particular reference to the time of calving, the duration of
pregnancy in ewes and the feeding program.
Biochemical examination to detect the presence of hypoglycemia,
ketonaemia and ketonuria are necessary to establish a diagnosis.
In laboratory findings you observe
 Reduction of blood glucose levels from the normal of 1528
 Ketosis Cont’d---
approximately 50 mg/deci liter to 20 - 40 mg/deci liter
 Hypoglycemia, ketonaemia and ketonuria are characteristic of the
disease.
It is possible to know a cow with ketosis by comparing with the
standard as
Normal cows have plasma BHBA concentrations less than
1000µmol/ liter of plasma
Cows with subclinical ketosis have concentrations greater than
1400 µmol/liter
Cows with clinical ketosis have concentrations often in excess of
2500 µmol/ liter
Milk and urine can be used as specimens to diagnose ketosis in
cows.
These tests have the advantage of being inexpensive, giving
immediate results.
A minor source of error is that the concentration of ketone bodies in
1529
these fluids will depend not only on the ketone level of the blood
 Ketosis Cont’d---
but also on the amount of urine excreted or on the milk yield.
However, milk is less variable, easier to collect and may give fewer
false negatives with subclinical ketosis.
Milk and urine ketone levels have been traditionally detected by the
reaction of acetone and acetoacetate with sodium nitroprusside.
The result can be interpreted in a semi-quantitative manner based
on the intensity of the reaction.
But nowadays, several products are available commercially as test
powders or strips.
They are commonly accompanied by a color chart that allows a
classification in grades such as negative, trace, small, moderate,
large, based on the intensity of the color of the reaction.
Differential diagnosis
There are many diseases of cattle and ewes those must be
differentiated from ketosis.
So that the wasting form of ketosis must be differentiated from
1530
 Ketosis Cont’d---
Abomasal displacement
Traumatic reticulitis
Primary indigestion
Cystitis and pyelonephritis
 Diabetes mellitus.
While the nervous form of ketosis must be differentiated from
Rabies
Hypomagnesaemia
Bovine spongiform encephalopathy
Treatment
The rational treatment in ketosis is
To relieve the need for glucose formation from tissues and
Allow ketone body utilization to continue normally
Theoretically, the simplest means of doing this is by the
administration of glucose replacement therapy.
Ideally, treatment should be at an early stage of the disease to
minimize loss with subclinical ketosis. 1531
 Ketosis Cont’d---
Treatment of ketosis can be done by
 1. Replacement therapy
 2. Hormonal therapy in combination with replacement therapy
1. Replacement therapy
Replacement therapy can be achieved by
1. IV injection of 500 ml of a 50% solution of glucose.
This results in transient hyperglycemia, increased insulin and
decreased glucagon secretion, and reduced plasma concentration
of non –esterified fatty acids.
2. IP injections of 20% solution of dextrose may be used
alternatively
But it is not widely used because it is accompanied by risk of
infection.
3. Other sugars, especially fructose, either alone or as a
mixture of glucose and fructose (invert sugar)have been used
in an effort to prolong the response
1532
 Ketosis Cont’d---
4.Perosadministration of propylene glycol and glycerine/glycerol
To overcome the necessity for repeated injections, propylene glycol
can be administered as a drench.
The traditional does is 225 gm twice daily for 2 days, followed by
110 gm daily for 2 days to cattle.
Propylene glycol (200-700 gm daily) or salts of propionic acid, can
be administered in the feed and give good results.
Administration in feed is preferred because this method avoids
dangers of aspiration with drenching.
5. Using of other glucose precursors
Lactates and propionates have glycogenic effect and can be used in
treatment of ketosis in cattle.
Some of lactates those are used in treatment of ketosis are
Sodium propionate
Sodium acetate
Calcium and sodium lactate 1533
 Ketosis Cont’d---
Ammonium lactate
Because of its glucogenic effect, sodium propionate is theoretically
a suitable treatment in ketosis.
So administer 110 - 225 gm doses daily, the response in cattle is
often very slow.
Lactates are also highly glucogenic .
But both calcium and sodium lactate (1 kg initially, followed by
0.5 kg for 7 days).
Sodium acetate in dose of 110-500 gm/day have given used in
treatment of ketosis.
But it has less satisfactory results than those obtained with sodium
propionate.
Ammonium lactate in dose of 200 gm for 5 days give a good result
in treatment of ketosis and has been used extensively.
2. Hormonal therapy in combination with replacement therapy
Hyperglycemia occurs within 24 h of administration of
1534
glucocorticoids.
 Ketosis Cont’d---
Hyperglycemia appears as the result from a repartitioning of glucose
in the body rather than from gluconeogenesis.
A hyperglycemic state is produced for 4 - 6 days in ketotic cows by
administration of
 Ten milligram( 10 mg) of dexamethasone 21-isonicotinate
Forty milligram(40mg)dexamethasone sodium phosphate
Five milligram(5 mg) flumethasone
So that it is better to use corticosteroids and IV glucose to treat
ketosis .
Because it is superior with fewer relapses than therapy with
corticosteroids or glucose alone.
In the early-onset cases of ketosis that are unresponsive to glucose
and/or corticosteroid therapy, it is possible to use insulin.
Insulin is administered in conjunction with either glucose or
glucocorticoids.
Because insulin facilitates
1535
 Ketosis Cont’d---
Cellular uptake of glucose
Suppression of fatty acid metabolism
 Stimulation of hepatic gluconeogenesis
Insulin can be administered in conjunction with either glucose or
glucocorticoids.
The common preparates is protamine zinc insulin given at the dose
of 200-300 IU SC per animal every 24 - 48 hours as required.
Vitamin B12 and cobalt are indicated in regions where cobalt
deficiency is a risk factor for ketosis.
Control and preventation measures
The control of clinical ketosis is integrally related to the adequate
nutrition of the cow in the dry and lactating period.
Cows should neither have been starved nor be overfat at calving.
Careful estimation of diets by reference to feed value tables is
recommended
Cows that are-housed should get some exercise each day .
In the herds where the disease is a particular problem during the1536
 Ketosis Cont’d---
stabling period, the cattle should be turned out to pasture as soon
as possible in the spring.
The ration should contain adequate amount of cobalt, phosphorus
and iodine.
If there is a high incidence in a herd receiving large quantities of
ensilage, reduction of the amount fed for a trial period is indicated.
If the incidence of ketosis is high energy supplements are
necessary.
Some of them are
Propylene glycol: - Propylene glycol has been drenched to cattle
in early lactation at doses varying from 350 to 1000 ml daily for 10
days after calving.
Glycerol: - It can replace propylene glycol in similar dose and is
given by drenching.
Propionic acid and its salts: -Propionic acid is absorbed across
the rumen wall and is transported to the liver where it is converted to
glucose via gluconeogenesis as the result it increase in serum blood 1537
 Ketosis Cont’d---
Ionophores : -It alters bacterial flora of the rumen, leading to
decreases in Gram-positive bacteria, protozoa, and fungi but it
increases the Gram-negative bacteria.
The net effect of these changes in bacterial flora is increased
propionate production and a decrease in acetate and butyrate
production providing increased gluconeogenic precursors.
It can be administered as a slow release capsule to cattle 2-4 weeks
before calving.
Niacin:- It is antilipolytic and induces increases in blood glucose
and insulin.
Niacin is given in the feed has a beneficial effect on subclinical
ketosis in cattle.
It has been suggested that it should be supplemented from 2 weeks
prior to parturition to 12 weeks post partum to prevent ketosis in
dairy herds.
To prevent ketosis herd monitoring is crucial.
Herd monitoring comprises 1538
 Ketosis Cont’d---
Biochemical monitoring of herds for subclinical ketosis.
It can be conducted using blood glucose estimations on a sample of
cows in their second week of lactation.
As the result blood glucose levels of below 35 mg/deciliter suggest
subclinical ketosis.
For individual cows, blood glucose estimations should be done at
about 14 days after calving.
More commonly, testing for ketones is done in urine or milk of cows
in their first or second week of lactation.
It is recommended for
For early detection of ketosis and
For early treatment to prevent milk loss and ketosis-associated
disease.

1539
7.3. Copper deficiency
It is also called hypocupporus which affects cattle, sheep and rarely
pigs and characterized by
Pica
Anemia
 Diarrhoea
Etiology
Copper deficiency can be primary or secondary.
Primary copper deficiency occurs due to inadequate levels of
cupper in the diet.
Secondary copper deficiency due to conditioning factors such as
excess molybdenum and sulfur in the diet.
Epidemiology
Primarily it is common in young pastured ruminants (cattle, sheep,
goats, and farmed deer) in spring and summer.
Primary deficiency occurs in sandy soil and heavily weathered
areas; secondary in peat or muck soil areas.
1540
Feed and water supplies may contain molybdenum, sulfate and iron
 Cupper Cont’d---
which interfere with copper metabolism.
It may be congenital in newborn lambs (swayback) if ewes deficient
or delayed in nursing lambs (enzootic ataxia).
Some breeds of sheep are highly susceptible.
Pathogenesis
In cupper deficiency the amount of cupper in plasma of blood
reduces and reach 0.2 mg/ liter( in normal animal 1 mg/liter).
As the result the biosynthesis of haemoglubin is disturbed and
develops anemia.
And the ability of blood to carry oxygen is reduced and in the
organism accumulates the intermediate product of metabolism and
develops acidosis.
As the result in animals develops
Diarrhea
Loss of production and growth rate
Cupper deficiency influences the biosynthesis of calcipherol and
keratinization of skin, hair and feather. 1541
 Cupper Cont’d---
That is why it is one of the etiology of osteodystrophy and changing
of the colour of hairs and feather.
Clinical signs
It may have as herd problem.
Young growing ruminants on pasture are more susceptible
Some of the common clinical signs are
Unthriftiness,
Changing of the hair color
Chronic diarrhea in molybdenosis (secondary deficiency),
Chronic lameness
 Neonatal ataxia
 Anemia in later stages of deficiency and called falling disease in
adult cattle.
Necropsy findings
The main necropsy findings are
Emaciation of the carcass
Anemia of the carcass
Hemosiderosis in liver and spleen
Osteodystrophy
Demyelination in enzootic ataxia
Myocardiopathy 1542
 Cupper Cont’d---
Diagnosis
Diagnose of cupper deficiency can be done based on history,
clinical signs and laboratory test.
Epidemiology, history and the presence of anemia in the herd can
help to suscepect cupper deficiency.
However, diagnostic confirmation can be done by
 Low serum and hepatic copper
 Having good response to treatment
Differential diagnosis
Copper deficiency must be differentiated from herd problems
associated with the following clinical findings.
Unthriftiness due to intestinal parasitism
Malnutrition due to energy-protein deficiency
Lameness caused by osteodystrophy due to calcium, phosphorus,
and vitamin D imbalance
 Anemia due to pediculosis
1543
Neonatal ataxia in lambs (congenital swayback and enzootic ataxia)
 Cupper Cont’d---
Cerebellar hypoplasia
Hypothermia
Meningitis
Treatment
The most common methods those are used to treat cupper
deficiency are
1. Copper sulfate orally
Oral dosing with 4 gm of copper sulfate for calves from 2 to 6
months of age and 8-10 gms for mature cattle given weekly for 3-5
weeks
2. Cupper glycinate parenterally
Parenterally administration of cupper glycinate at the dose of in
beef herds is 120 mg of copper for adult cattle and 60 mg of copper
for calves is useful.
One dose of copper glycinate will maintain adequate copper levels
for about 60 - 90 days.
1544
 Cupper Cont’d---
Control and preventation measures
We can control the disease by oral dosing or dietary
supplementation salts of cupper like copper oxide in feed or on
pasture.
Parenteral administration of copper salt solutions are at strategic
times and are also important.
Other methods those are used to prevent cupper deficiency are
 Genetic selection of breeds which are resistance of cupper
deficiency
 Removal of sulfates from water supply
7.4. Cobalt deficiency
It is the disease of mostly ruminant animals caused by ingesting a
diet deficient in cobalt, which is required for the synthesis of
vitamin B12.
It is caused mainly by low content of cobalt in the soil and plants
(below 2 mg/ kg).
1545
 Cobalt Cont’d---
This condition predispose the animals to the development of endemic
disease called hypocobaltosis(absence of cobalt A cobaltosis).
Cobalt was first shown to be an essential nutrient for sheep and cattle in
1930.
Because it causes the two naturally occurring diseases called 'coast
disease' of sheep, and 'wasting disease‘ or enzootic marasmus, of cattle.
The disease is characterized clinically by inappetance and loss of body
weight.
It has also some effects on reproductive performance of sheep.
Etiology
The disease is caused by a deficiency of cobalt in the
diet which results in a deficiency of vitamin B12.

1546
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