FISH DISEASES

&
HEALTH MANAGEMENT

Ravi Kant Verma

V P Saini
Tejpal Dahiya
Pawan Kumar Yadav
 FISH DISEASES
&
HEALTH MANAGEMENT

Ravi Kant Verma

V P Saini
Tejpal Dahiya
Pawan Kumar Yadav

Agrotech Publishing Academy

Udaipur

Published by:
Mrs Geeta Somani

Agrotech Publishing Academy

1-G-24, sec. 5 (Gaytri nagar)

Hiran Magri, Udaipur-313001 (India)

Phone: 0294-2465635

First Edition 2010

© 2010 by Authors

All Right Reserved

ISBN…

Printed at…

Preface
Fisheries sector occupies a very important place in the socio-
economic development of the country. It has been recognized as a
powerful income and employment generator as it stimulates
growth of a number of subsidiary industries and is a source of
cheap and nutritious food besides being a foreign exchange earner.
Most importantly, it is the source of livelihood for a large section
of economically backward population of the country.

The book entitled “Perspectives in Aquaculture” has been

written with the intention to meet the needs of fisheries graduate
and post-graduate students working on various aspects of
aquaculture especially fish diseases. In this book, concise
descriptions of the topics, introduction to fresh water aquaculture,
importance of soil and water in fish health management, fish health
management, an introduction to bacterial fish, prawn and cat fish
diseases in India, fish disease control, use of probiotic in fish
disease abatement, fish immunology and status of fish vaccines in
aquaculture that is suitable for teaching the fundamental concepts
of fisheries. We are sure that this book will serve the purpose of
research work for beginners and entrepreneurs, who would like to
take up fish culture. This may also serve as a useful tool for the
students and researcher in understanding intricacies of aquatic
ecosystem and effect on fish health.

We acknowledge our gratitude to the authors of different

books and research papers, which were used as reference tool in
the preparation of this book. We extend our hearty thanks to Dr.
R.C. Sihag (Professor, Zoology and Aquaculture, CCS HAU,
Hisar), Dr. Sunil Kumar Saini (Asstt. Professor, MPUAT, Udaipur,
Rajasthan) for all needed co-operation and moral support.
Authors
 CONTENTS

Preface

About the authors

About the book

List of contributers

An introduction to fish health management

Importance of soil and water in fish health

management

Bacterial diseases of fresh water fishes

An introduction to bacterial fish diseases in India

Diseases of catfish and their control

An introduction to fresh water prawn diseases

Prominent catfish diseases

Probiotics in aquaculture

Fish immunology

Status of Fish Vaccines

 About The Authors

Ravi Kant Verma (CSIR NET-JRF) is research scholar in NRCE,

Hisar, (Haryana). He did his M.Sc. in 2006 and pursuing his
Ph.D. in same department. He has published several research
publications on fish biotechnology and pathology in reputed
journals. He is the ordinary member of SOFTI Cochin. His fields
of interest are fish biotechnology, pathology, immunology and
fish health management.

Dr. Tejpal Dahiya (RGNF- fellow) is lecturer in Department of

Zoology, Kurukhetra University Kurukhetra He did his M.Sc. in
2005 and Ph.D. Degree in 2010 from CCS Haryana Agricultural
University, Hisar, (Haryana). He has an extensive and rich
experience in field of fish pathology and teaching. He has
published several research publications on fish pathology in
reputed journals.

Pawan Kumar Yadav (CSIR NET-JRF) is lecturer in Department

of Biotechnology, Dr. K. N. M. I. P. E. R., Modinagar (U. P.). He
did his M.Sc. in 2005 from M.D. University, Rohtak, (Haryana)
and pursuing his Ph.D. in Genetics from Department of
Genetics, CCS Haryana Agricultural University, Hisar. He has
published several research publications and articles in various
national and international journals of repute. He has an
 extensive and rich experience of 3 years to his account in field
of teaching and research.

About The Book

The technology of culturing fish has attracted investors to
the aquaculture industry. More often, however, the
significant impact of disease outbreaks is overlooked. It is
only when occurrence of mortalities, retarded growth,
reduced fecundity or lesioned fish alarmingly reduces profit
that the fish farmer become aware of disease
problems.Unfortunately, the disease may be at an advanced
stage by the time diagnosis is attempted.

Aquaculture production in Asia has grown rapidly.

Unfortunately, a number of infectious disease have emerged
as well owing to deteriorating conditions in farms not
practicing sound culture management.To aid the industry in
dealing with its problems in diseases, the present book“Fish
Diseases and Health Management”is prepared.This book
provides ten research articles of reputed researchers in the
field of fish health management. Articles cover topics of
environmental, bacterial,fungal and viral diseases.Beside,
these the three most important and new topics –(i)
Probiotics in aquaculture(ii) Fish immunology & (iii) Status
of Fish Vaccines are the main attraction of the book.
The objective of this book is to acquaint the fish farmers &
students with the basic knowledge on disease causation
and development among cultured fish and the management
of major fish diseases.This book will be very useful for UG
and PG students, personal and research libraries because
it gives current literature of many aspects of fish health
management in one volume.
 List Of Contributors

1. Sihag, R.C., Department of Zoology & Aquaculture,

CCS Haryana Agricultural University, Hisar, Haryana
- 125004, India.
2. Dahiya, Tejpal, Department of Zoology, Kurukhertra
University, Kurukhertra.
3. Verma, Ravi Kant, Department of Zoology &
Aquaculture, CCS Haryana Agricultural University,
Hisar, Haryana - 125004, India.
4. Vijayanti Jakhar, Department of Zoology &
Aquaculture, CCS Haryana Agricultural University,
Hisar, Haryana - 125004, India.
5. Gupta, Manisha, Department of Zoology &
Aquaculture, CCS Haryana Agricultural University,
Hisar, Haryana - 125004, India.
6. Sharma, Mukta, Department of Zoology &
Aquaculture, CCS Haryana Agricultural University,
Hisar, Haryana - 125004, India.
7. Yadav, Pawan Kumar, Department of Biotechnology, Dr. K.N.
Modi institute of pharmaceutical education and research,
Modi Nagar, Ghaziabad, U.P.
8. Saini, V. P.

An Introduction To Fish Health

Management

Aquaculture is the production of fish and shellfish for

market under controlled or semi-controlled conditions.
During the past three decades the raising of fish for human
consumption has been increasing dramatically especially in
the sociologically and industrially developed countries. The
increased production has come about largely as a result of
consumer awareness that fish and shellfish are
nutritionally beneficial to health. Unfortunately, the
production increase in some countries has been so great
that the supply of propagated food fish has exceeded the
demand in the marketplace. Attendant to the increased
production of food fish under intensive culture conditions
has been the increased loss of production potential through
infectious and noninfectious disease processes. In many
cases the episodes have been so severe that 45-55% of the
numbers of fish at the beginning of the rearing process
have died before they became ready for market.
 For commercial success, an aquaculture operation
must maintain fish at densities that greatly exceed those
normally found in nature. Under these conditions, fish
must not only survive, but also grow rapidly. Regardless of
the culture system used (e.g., ponds, raceways, reuse
systems, cages) it is imperative that the culturist maintain
an environment conducive to good fish health. A wide
variety of parasites and pathogens can and do infect fish.
Most disease agents are naturally present in low numbers
and normally do not cause problems. The natural defense
mechanisms of fish (i.e. undamaged skin, mucus covering
the skin, and various components of the immune system)
keep disease agents in check. However, when fish already
crowded in culture operations are further stressed (e.g., by
low dissolved oxygen, nutritionally inadequate feeds,
excessive handling) their natural disease defense systems
may be weakened and the ability of the fish to protect itself
against infectious diseases maybe reduced. Disease
induced catastrophic mortalities are frequently the result
of, and response to, a stressful experience. Fish producers
today have very limited choices for controlling fish disease
problems.
A fish health prevention/maintenance program is
presently recognized as a required management practice by
fish producers to lessen the risk of diseases. This
management approach has become particularly critical for
producers using recirculating systems. Fish health
prevention/maintenance programs require many elements
including quarantine procedures, examination and
monitoring for fish pathogens, and prophylactic treatments
for parasites. Developing fish health prevention protocols
are necessary to reduce likelihood of bacterial and parasitic
diseases.
Fish health management is a term used in
aquaculture to describe management practices which are
 designed to prevent fish diseases. Successful fish health
management begins with prevention of disease rather than
treatment. Management practices directed at limiting stress
are likely to be most effective in preventing disease
outbreaks.

Any manager of fish health seeks to achieve two goals:

 To maximize immunocompetence in fish populations; and
 To reduce or eliminate potential pathogens (i.e., parasites,
bacteria, viruses and fungi) in culture systems.
EXTRINSIC FACTORS AFFECTING FISH HEALTH
The extrinsic or environmental factors which are known to
compromise the health status
of fish individually or collectively can be grouped into the
following categories according to their location within the
system:
(1) water-associated;
(2) pond-associated;
(3) nutrition-associated;
(4) management-associated.
(1) Water-associated factors:
Among water-related factors identified as affecting the
productivity of aquaculture systems, water temperature and
dissolved oxygen content have the most significant effects
on fish health. They are inherent in all water supplies and
are subject to fluctuations to which the fish in the system
must adapt. The physiological effects of the fluctuations are
very broad, ranging from a change in metabolic rate to
altering the susceptibility to pathogens. In this regard, the
documentation of environmental changes and the
occurrence of infectious and noninfectious disease
processes could be an invaluable aid in predicting the
likelihood of a subsequent disease episode.
(2) Pond-associated factors:
The primary effect here is requiring a particular fish to live
in a pond configuration which does not meet its behavioral
requirements.

(3) Nutrition-associated factors:

One would think that in this day and age of high quality
commercial diets, nutritional problems would not occur,
but they do, all too frequently. Growth rate is a very reliable
indicator of the health of the population. Deviations of as
little as 1% from the expected growth rate can be measured
quite accurately and evaluated with a high degree of
statistical validity. Over feeding a population is another
health-compromising situation. In this case there are
frequently abnormal amounts of abdominal fat and hepatic
glycogen deposits. The effects are not often seen
immediately, but can be implicated in the milieu of casual
factors of an unhealthy state.
(4) Management-associated factors:
The primary health-threatening factors in this category
emanate from exceeding one or more of the pond carrying
capacities, inadequate housekeeping practices, inadequate
record-keeping practices, and undue physical stressors.

INTRINSIC FACTORS AFFECTING FISH HEALTH

The intrinsic or somatic factors originate within the fish
itself. They are largely governed
by the genetic make-up which by and large dictates the
physiological and psychological responses of the fish to the
extrinsic factors.
Perhaps the major intrinsic factor which fish health
managers have some control over is
the generation of endogenous ammonia . It is well known
that free ammonia in the system is deleterious to the health
of fish when specified limits are exceeded. The main impact
is on the gill lamellae in the form of epithelial hypertrophy
and hyperplasia which reduce the oxygen uptake by the
fish, thus impacting the physiological well-being of the fish.
The control of this process is to reduce the dietary protein,
or to decrease the retention time of ammonia in the pond by
increasing the water flow, or to reduce the population of
fish in the pond, or to reduce the water temperature, or to
decrease the pH of the system.
A second intrinsic factor over which some control
can be exercised is the healthy or
chronic asymptomatic carrier of infectious agents. It is
apparent from studies with Renibacterium salmoninarum
and Aeromonas salmonicida carriers that the presence of
these bacteria within the fish negatively impact their growth
potential and present a measurable threat to the uninfected
portion of the population.
STRESS

Most fish can tolerate environmental conditions that differ

somewhat from the natural conditions in which they
evolved. This does not mean, however, that they will be as
healthy or live their full normal life span. For example,
keeping a fish in water that is cooler (or warmer) than its
preferred condition forces its body organs to work harder to
keep it alive. That is, such conditions place the fish under
increased stress.

Stress is any condition that causes physical or

mental discomfort that result in the release of stress-related
hormones or results in specific physiological responses. For
example, stressful events will cause an increase in heart
rate, blood pressure, increased blood sugar, and the release
of cortisol. Stress can be physical, psychological, or
environmental. Stress can either be short and sudden, or
long and chronic. Mild, short-term stress has few serious
health effects, but long-term stress or severe, short-term
stress contribute to many of the illnesses and deaths in
aquarium fish. Increased stress reduces a fish's ability to
ward off diseases and heal itself (e.g., if its fins get nicked,
or parasites get introduced into the tank with newly
purchased fish). In addition, stress reduces a fish's ability
to breed successfully and shortens its natural life span.
Thus, one of the most important goals of a fish keeper is to
remove sources of stress wherever possible.

The causes of stress

There are dozens of potential stresses to fish but some of

the more common causes are:

 Elevated  Inadequate
ammonia tank size
 Elevated  Overstocking
nitrate of tank
 Improper pH  Medications
level and water
 Fluctuations treatments
in  Improper
temperature nutrition
 Improper  Disturbance
salinity of the tank
 Low oxygen  Harvesting
levels and shipping
 Harassment of fish
from other  Lack of
fish enough fish to
 Lack of hiding provide
places schooling
Elevated ammonia, nitrite, and nitrate all create
deterioration in fish health due to stress. High levels can
cause severe stress, whereas slightly elevated levels can
contribute to chronic stress.

pH levels that change abruptly cause acute stress and

continually elevated or lowered pH levels can cause chronic
stress. Many fish adapt to long-term changes, but there are
limits. pH changes of more than 1.5 points below or above
recommended levels have a negative effect over time and
should never be considered acceptable.

Temperature fluctuations are a much underappreciated

stressor of fish. Most tropical freshwater and marine fish do
not tolerate temperature changes very well. Many tanks
that are not set up properly will have over the
recommended maximum of one degree of temperature
fluctuation in a 24-hour period due to room temperature,
lights, and equipment. The daily fluctuations create
chronic.

Wild fish live within very specific salinity levels (levels of

salt in the water). Their bodies work hard to maintain the
osmotic gradient between themselves and their
environment. If their environmental salinity is not specific
to their needs and is not held at a steady level, they have to
work harder to maintain their osmotic gradient, which
generates chronic stress.

Oxygen levels that are below recommended levels can cause

fish to 'breathe' faster than optimum and this can result in
chronic stress. Obviously, very low oxygen levels can lead to
severe short-term stress and death.
Harassment from other fish and lack of hiding spaces go
hand in hand. There should be two suitable hiding spaces
for every fish in the tank, otherwise, there are going to be
fish that are stressed and bullied. Unlike their environment
in the wild, these fish are confined and cannot get away
from aggressors. Aggression is a very real problem in many
tanks that leads to many injuries, infections, and death.

Overstocking of the tank is a common problem that

contributes to almost all of the stresses in the above list,
from water pollution to oxygen depletion to harassment.
Tanks should not be overstocked.

Improper nutrition is also a commonly overlooked stressor

of fish. Many fish can live on minimal nutrition with old or
stale flake foods, but this poor nutrition is a chronic stress.
A variety of well-preserved dry foods as well as freeze-dried,
fresh, and frozen foods specifically designed for individual
species are necessary to prevent chronic nutritional stress.

Disturbing the tank through banging on the glass,

constantly netting fish, or rearranging décor stresses fish
and should be kept to the necessary minimum.

Thus it can be seen that many factors lead to fish

stress. Minimizing and eliminating sources of stress
increases the chances of keeping tank inhabitants healthy.
The exact amount of stress an individual fish can take
depends greatly on what species it is, its age and size, etc. A
stressed fish is a weakened fish. Although it may appear
healthy to the casual observer, it will be more susceptible to
disease, injury, etc. In contrast, healthy (unstressed) fish
will be able to ward off disease and infection on their own.
Thus, the appearance of disease in a tank is frequently
brought on by ``poor water conditions'' that leave fish with
weakened immune systems.

Common symptoms of stress include:

 Fish stays near the surface gasping for breath,

indicating that it has trouble getting enough oxygen
(the concentration of dissolved oxygen is highest near
the water's surface). Possible causes include low
oxygen concentration due to poor water circulation,
toxins that have damaged its gills, high ammonia or
nitrite levels, etc.
 Fish won't eat, or doesn't eat as aggressively as in
past.
 Fish stays hidden continuously and won't come out
where it can be seen. Possible causes: aggressive
fish, insufficient cover (e.g., plants, wood, etc.) to
make fish feel ``safe'' while swimming about.
 Fish has nicked fins, open wounds that don't seem to
heal. Possible cause: fish is target of aggression.
Normally, minor nicks and cuts heal quickly. If they
don't, stress levels may be suppressing the fish's
immune system.
 Fish has disease (parasites, fungus, etc.) Of course,
the disease itself is a major problem. But in most
cases, a healthy fish's immune system keeps it from
getting sick in the first place. Thus, getting sick is a
sign that the fish is in a stressed state (or had been
until recently).

Effect of stress on a fish's health

Short-term stress will cause an increase in heart

rate, blood pressure, and respiration. The fish can only
maintain these altered states for a short period of time and
then they will adapt or the stress will become chronic.
Stress is accompanied by the release of the hormone
cortisol, which is responsible for many of the negative
health effects associated with stress. In addition to having a
negative effect on growth, reproduction, and digestion,
chronic stress will also lower the ability of the immune
system to respond effectively and fully. This lowered
immune response is what allows parasites, bacteria, and
fungi to infect a stressed fish and cause disease and death.

Stress in fish caused by physical disturbances

encountered in aquaculture, such as handling and
transport, evokes a variety of responses that may be
adaptive or maladaptive. The overall effect of stress may be
considered as a change in biological condition beyond the
normal resting state that challenges homeostasis and, thus,
presents a threat to the fish's health. These stress induced
changes are grouped as primary; secondary, which includes
metabolic, haematological, hydromineral and structural;
and animal' responses. Many of these responses can be
used as quantitative indicators of stress. A major focus of
current research is on the response of the hypothalamic-
pituitary-interrenal axis and the resultant elevation of
circulating corticosteroids.

Stress, through the action of corticosteroids, may:

a) Reduce immunocompetence by influencing

lymphocyte numbers and antibody- production capacity;
and

b) Affect reproduction by altering levels and patterns

of reproductive hormones that influence maturation.
 Stress may also alter metabolic scope in fish and
affect growth, partly as a result of the catabolic or
gluconeogenic effect of corticosteroids. Although certain
stressors encountered during normal aquaculture
procedures may be unavoidable, a number of practical
approaches are suggested that would help to alleviate the
detrimental effects of stress in fish.

Types of Fish Diseases

Disease is rarely a simple association between a pathogen

and a host fish. Usually other circumstances must be
present for active disease to develop in a population. These
circumstances are generally grouped under the umbrella
term "Stress".
Figure . Disease rarely results from simple contact
between the fish and a potential pathogen.
Environmental problems, such as poor water quality, or
other stressors often contribute to the outbreak of
disease

There are two broad categories of diseases that occur in

fish.
1. Infectious diseases are contagious diseases caused
by parasites, bacteria, viruses, or fungi. These often
require some type of medication to help the fish
recover.
2. Non-infectious diseases are broadly categorized as
environmental, nutritional, or genetic. These problems
are often corrected by changing management
practices.

Infectious diseases.
Infectious diseases are broadly categorized as
parasitic, bacterial, viral, or fungal diseases.
Parasitic diseases
Parasitic diseases of fish are most frequently caused
by small microscopic organisms called protozoa which live
in the aquatic environment. There are a variety of
protozoans which infest the gills and skin of fish causing
irritation, weight loss, and eventually death.
Icthyophthiriasis and costiatis are examples of diseases
caused in fish by protozoans. Most protozoan infections are
relatively easy to control using standard fishery chemicals
such as copper sulfate, formalin, or potassium
permanganate.

Bacterial diseases.
Typically a fish infected with a bacterial disease will
have hemorrhagic spots or ulcers along the body wall and
around the eyes and mouth. They may also have an
enlarged, fluid-filled abdomen, and protruding eyes.
Bacterial diseases can also be external, resulting in erosion
of skin and ulceration. Columnaris is an example of an
external bacterial infection which may be caused by rough
handling. Some of the common bacterial diseases that
occur in fish are dropsy, bacterial gill disease, vibriosis and
tail and fin rot. Bacterial infections require treatment with
medicated feeds containing antibiotics which are approved
for use in fish by the Food and Drug Administration.

Viral diseases

Viral diseases are impossible to distinguish from

bacterial diseases without special laboratory tests. The
most important viral infection which affects fish production
is Channel Catfish Virus Disease, caused by a herpes virus.
Examples of diseases caused by virus in fish include
Infectious Pancreatic Necrosis (IPN), Viral Hemorrhagic
septicemia and Infectious Hematopoietic Necrosis (IHN).
Consultation with an aquaculture or fish health specialist is
recommended if a bacterial or viral disease is suspected for
killing of fish.

Fungal diseases
Fungal diseases are the fourth type of infectious
disease. Fungal spores are common in the aquatic
environment, but do not usually cause disease in healthy
fish. When fish are infected with an external parasite,
bacterial infection, or injured by handling, the fungi can
colonize damaged tissue on the exterior of the fish. These
areas appear to have a cottony growth or may appear as
brown matted areas when the fish are removed from the
water. Saprolegniasis, Branchiomycosis and
Ichthyosporidiosis are commonly occurring diseases caused
 by fungus in fish.Formalin or potassium permanganate are
effective against most fungal infections. Since fungi are
usually a secondary problem it is important to diagnose the
original problem and correct it as well.

Pathogen Transmission

Pathogens can be transmitted several ways in a

recirculating system:

 in the water
 fish to fish
 by vectors and fomites
 in the food
Introduction of water used to ship fish can be a key source
of pathogens. Shipping water often contains high numbers
of bacteria and may also contain parasites or other
pathogens. These organisms are easily transferred from
tank to tank in the recirculating water or by aerosolization
(in mist or spray) of water from one tank or system to
another.

Within a single tank pathogens can be spread directly from

fish to fish. Higher stocking densities and increased fish-to-
fish contact (as seen in aggressive species) can increase the
rate of spread of pathogens.

Vectors

Vectors are organisms that can transmit disease-causing

organisms from one animal to another. For example, the
crustacean parasite Argulus (“fish louse”) causes damage by
itself, but it is also believed to transmit bacteria and viruses
between fish. Leeches are another vector that can transmit
blood-borne parasites and bacteria between fish.
Additionally, people can act as vectors by transmitting
water and pathogens from one tank to another via their
hands or arms.

Fomites

Fomites are inanimate objects that can transmit

diseases. Examples of fomites in aquaculture systems
include equipment, such as nets and siphon hoses that are
not properly disinfected before being used in other tanks or
vats.

Food

Food can also be a source of disease. Frozen and live

foods can transmit bacteria, parasites, viruses, and fungi.
In addition, feeds that have been improperly stored can
contain pathogenic bacteria or mycotoxins, dangerous
chemicals produced by the growth of certain types of fungi
in the feed.

Non-infectious diseases
Non-infectious diseases can be broadly categorized
as environmental, nutritional, or genetic.
Environmental diseases are the most important in
commercial aquaculture. Environmental diseases include
low dissolved oxygen, high ammonia, high nitrite or natural
or man-made toxins in the aquatic environment. Proper
techniques of managing water quality enable producers to
prevent most environmental diseases.
Nutritional diseases can be very difficult to diagnose. A
classic example of a nutritional disease of catfish is "broken
back disease," caused by vitamin C deficiency. The lack of
dietary vitamin C contributes to improper bone
development, resulting in deformation of the spinal column.
Another important nutritional disease of catfish is "no blood
disease" which may be related to a folic acid deficiency.
Affected fish become anemic and may die. The condition
seems to disappear when the deficient feed is discarded and
a new feed provided.
Genetic abnormalities include conformational oddities such
as lack of a tail or presence of an extra tail. Most of these
are of minimal significance; however, it is important to
bring in unrelated fish for use as broodstock every few
years to minimize inbreeding.

Preventative Medicine Principles

Preventing disease by following the essential

principles of fish health management is the most cost
effective management scheme for aquaculture systems.
The principles include understanding and attending to:
1) Species biology, including all known and important
genetic considerations;
2) Nutrition;
3) Water quality;
4) Quarantine;
5) Sanitation; and
6) Disinfection.

Species biology
A basic knowledge of the anatomy, physiology,
behavior, genetics, social interactions and environmental
needs of cultured species is critical to proper management.
Some fish, such as cichlids, may be very aggressive toward
each other, and they may even eat smaller members of their
species. Some species may require a culture system with a
large surface area rather than a large volume (e.g., four-
eyed fish, Anableps). Other species may require specific
substrates or habitats. For example, shell-dwelling cichlids
fare best with shells or pipes in the tank.
The genetics of many species are not well known, and their
effects may be subtle. However, some dramatic diseases
have a known genetic cause and in these cases, genetic
selection is important when choosing potential broodstock.
Some of these diseases include: “saddleback” (abnormal
dorsal fin or death) in tilapia, a condition controlled by a
single gene; poor survivability among black angelfish
offspring; early death of offspring from crosses of koi with
certain scale patterns (the result of two different genes
interacting); and the potential for inbreeding depression.
Inbreeding depression refers to a reduction in things such
as survivability, desirable traits (such as size and color), or
overall health resulting from many generations of brother to
sister or parent to offspring crosses. Similarly, proper
broodstock selection is important to avoid breeding out of a
population genes that result in desirable properties (e.g.,
larger size, long fins, color, and scale type).
Nutrition
Nutritional considerations include size, palatability
(“attractiveness by taste”) and quantity of pellets, granules
or live foods; frequency of feeding, lighting (some species
may prefer to eat during the day, at night, or at “dusk”),
optimal temperature (species have a preferred temperature
for feeding, or may have reduced requirements when
temperatures are lower); as well as the bioavailability,
quality and quantity of micronutrients (vitamins, minerals)
and macronutrients (protein, fat, carbohydrate) in the diet.
Fish raised in outdoor ponds frequently benefit from live
foods present, but these nutritional sources are not
available in most recirculating systems). Consequently,
nutritionally-complete diets are essential for fish reared in
intensive recirculating systems. Unfortunately, the details
of what constitutes a nutritionally-complete diet are not
known for all species. Gross deformities in several species
of fish have been reported from production facilities using
recirculating systems. In particular, the cyprinids may grow
slowly and become particularly susceptible to
musculoskeletal and opercular abnormalities. Cyprinids
lack a true acid-producing stomach, so they cannot digest
certain forms of phosphorus, including tribasic calcium
phosphate and hydroxyapatite, derived from bone and
scales, which may contribute to abnormalities in these fish
due to the fishs inability to break down and use
phosphorus from these compounds. More soluble forms of
phosphorus, such as monobasic phosphates of potassium,
sodium, or calcium, are recommended.
Water quality
First and foremost, water quality must be compatible
with the requirements of the fish being held, especially with
regard to ammonia, nitrite, nitrate (in marine systems), pH,
temperature, dissolved oxygen, hardness, alkalinity, and
salinity. Although initial water quality in a system will be
determined by water source and water treatment regimes,
the long-term water quality in a aquaculture system
depends on numerous factors. The most important
considerations are the source of water in the system, fish
load, feeding rates, and biofilter capacity.
Water from different sources may have different
potential problems that must be addressed. Municipal
water may contain chlorines or chloramines; well water may
contain hydrogen sulfide, supersaturated gases (such as
nitrogen gas, resulting in gas bubble disease), high carbon
dioxide levels, and low oxygen levels, or high dissolved iron
levels, (all conditions that can be lethal to fish if not
corrected); surface water sources may be high in bacteria or
toxic chemicals resulting from runoff.
The most common water quality problems in aquaculture
systems are toxic levels of ammonia or nitrite caused by
imbalances between the capacity of the biofilter and the fish
load and feeding rates. This problem often occurs during
 the start up of a system, although it may occur at any time.
The bacteria in the biofilter can require three to eight weeks
to cycle (i.e., become established) at 25–27°C (77–81°F) and
even more time may be required at cooler temperatures.
Aquaculturists begin this cycling process prior to the
addition of fish by one of the following methods:
 adding ammonia directly into the system;
 adding a species of fish (such as black tetras,
Gymnocorymbus ternetzi) that is more tolerant of the
initial high ammonia and nitrite levels before adding the
final species to be held; or
 seeding the system/biofilter with bacteria from a
“healthy” established system or with bacteria from a
reputable commercial source.
In established filters, toxic levels of ammonia and nitrite
may result from overfeeding, crowding, or inefficient
removal of solids (such as feces and uneaten food),
resulting in breakdown of large quantities of proteins into
ammonia.
However, in addition to problems caused by source
water issues, ammonia, and nitrite (described above),
problems can also result from changes in water quality
parameters that were previously acceptable. Parameters
that can change over time in a system include dissolved
oxygen (DO, decreases), alkalinity (decreases), carbon
dioxide (can increase), and pH (decreases).
Low DO can occur during operation of a system as the
result of many different causes. Some of these include: high
stocking densities, inadequate water flow, inadequate
aeration, high organic loads in the system that lead to large
numbers of bacteria in addition to those in the biofilter,
high feeding rates or the use of certain chemicals such as
formalin.
 Some management options to prevent this drop in alkalinity
and pH and the potential rise in ammonia and nitrite
include:
 routine (once a week to once a month, depending upon
stocking density) measurement of alkalinity and pH, in
addition to other parameters mentioned previously;
 partial water changes on a routine basis (amount
dependent upon drops in alkalinity and pH over time) as
long as the source water has an adequate amount of
bicarbonate/carbonate (100 mg/L or more); and
 Adding sodium bicarbonate (baking soda) buffer as
needed.
Other important water quality parameters that may
require monitoring include hardness, salinity, organics, and
conductivity. These parameters may increase over time in a
system that does not undergo routine water changes, but is
only “topped off.” In such a situation these parameters may
reach levels that are not desirable for maintenance or
reproduction of some fish species.
Heavy metals, such as copper, zinc, and lead, are also toxic
to fish and may be present in the water source. If they are,
methods to remove them should be considered and system
levels should be checked regularly. Additionally, heavy
metals (such as copper or zinc) may be part of the hardware
of the system (though this is highly undesirable) and may
begin to leach into the water as the pH becomes more
acidic.

Quarantine
Quarantine is voluntary or compulsory isolation,
typically to prevent the spread of something considered
dangerous, often but not always disease. Quarantine is a
standard procedure in public aquaria because pathogens
can be introduced with new specimens. These potential
disease-causing organisms can result in major disease
problems in an established system. Ideally, quarantine
protocols should be in place and followed when fish are
transferred between facilities, such as when new
broodstock are brought into a facility. Adding new fish to an
established system without proper quarantine can spread
disease from the newly introduced fish to the established
stock or from the established stock (which may be healthy,
but may be a “carrier” of pathogens) to the newly
introduced fish.

Harvesting, shipping, transport, change of diet, and

other procedures result in immune system depression. As a
result, new fish may be more prone to shedding disease-
causing organisms (a lowered immune system results in
greater chance for infection and shedding) and infecting
other fish or they may be more susceptible to disease from
other fish. Quarantine allows new fish to become
acclimated to feeds, water quality, and husbandry
protocols, without the added stressors of other fish and
pathogens. At the same time, quarantine allows their
immune systems time to return to normal so that they can
combat diseases they may have developed during these
changes.
Following quarantine, especially with very valuable
fish, one suggested protocol is to place one or more fish
from the established stock with one or more fish from the
new group for a week or two to determine if there is any
lingering disease.
Quarantine protocols should contain the following elements
to minimize the potential spread of infectious organisms
and to allow the immune systems of new fish to stabilize.
All-in, all-out stocking.
Bringing fish in as a group from only one population
and keeping these animals together, or all-in, all-out
stocking, greatly reduces the potential for exposure to
pathogens that are not currently found at some level within
that population (i.e., potential cross-contamination from a
different group). Ideally, all-in, all-out quarantine involves
an entire system, room or facility. No new fish should be
added to a population that is currently under quarantine.
In some ways, isolation or separation resembles and
serves the same purpose as all-in, all-out stocking
preventing spread of potential pathogens from one group to
another. However, this procedure emphasizes separation or
isolation for different populations that must inhabit the
same facility where all-in, all-out stocking is not feasible. All
fish in quarantine at a specific time should be isolated or
separated from other populations at the facility, preferably
in a separate area with dedicated equipment.
Total time spent in quarantine or isolation may vary
according to the species of fish; results of sampling for
pathogens; commonly found pathogens, their life cycle, and
required treatment protocols; potential risks; costs; and
space and time limitations. For some retail facilities, this
may mean a separate tank system or individual tanks for
incoming shipments (especially for species that are typically
problematic). Dedicated equipment (nets, siphon hoses,
etc.) should be available for each “quarantine” system, or at
the very least, an effective disinfectant should be used for
equipment shared between systems.
Observation and diet adjustment.
Any acclimation protocol will benefit from
observation and diet adjustment. Observations of normal
and abnormal behaviors can provide information on the
status of the fish. Diet adjustment means gradual increase
over time in the percentage of the final diet fed until fish are
readily taking 100% of the final diet. A high quality diet
should be fed at all times during the transition, taking into
account species requirements.
 As fish are being acclimated to a new diet daily
observations should be made to assess feeding, observe
normal behavior, and identify potential disease problems.
Making sure that fish readily eat a manufactured feed
makes it easier to treat them with antibiotics because the
antibiotics can be added to the feed if necessary. It also
provides a stable, balanced base diet to which additional
elements can be added as necessary.
Sampling and treatment.
When feasible, fish that are under quarantine, or
that have been recently brought into a facility, should be
sampled or screened to determine the presence of
pathogens (i.e., parasites, bacteria, fungi and viruses) prior
to their introduction into a system and before any
treatment is started. Determining what disease-causing
organisms are present allows aquaculturists to be more
careful and scientific in their use of treatments and
preventative chemicals or antibiotics.
By sampling fish early on for the presence of pathogens or
accompanying diseases, scientifically-based treatments can
be applied before significant losses occur. At the very least,
fish brought into a multi-tank or multi-vat system with
other fish should be sampled for external parasites.
If necessary, sampling can be performed on live specimens.
External samples usually include taking a skin scrape, fin
clip and gill clip and examining these tissues under a
microscope. If fish cannot be sacrificed, examination of
fresh feces may provide information on parasite loads in the
gastrointestinal tract.
However, internal examination, which requires the sacrifice
of several specimens, is recommended if circumstances
allow. Full necropsies (sacrificing fish and carefully
examining external and internal tissues with and without
the use of a microscope) including microbiology should be
performed to rule out the presence of bacterial, viral and
fungal diseases, as well as to identify internal parasites.
Prior to this approach, however, consultation with a fish
health specialist is strongly recommended.
Reduction or elimination of infectious pathogens.
Introduction of known infectious pathogens should
be avoided or at the very least minimized. Preventative
treatments (“prophylactic” treatments), such as freshwater
dips for marine fish or salt water dips for freshwater fish,
can be helpful in removing initial loads of external
parasites. Use of external parasite treatments, such as
formalin or copper may be necessary if sampling
demonstrates the presence of parasites susceptible to those
treatments. A good clinical history of the fish and
information on their origin and possible diseases should be
obtained in order to help the aquaculturist and the fish
health specialist target their search for possible causes of
infectious and non-infectious diseases.
Wholesale and retail operations, especially those
working with large numbers of valuable fish, or koi
hobbyists with long-term, permanent populations, may add
one other step. They should consider combining a few
members of the “old” or “resident” population with a few
members of the “new” population in another system for a
week or two at the end of the quarantine period. This
approach will help determine if any subclinical infections
are present (subclinical means that a very low level of
disease is present but not causing obvious symptoms). If
this is the case, these diseased fish present in the resident
population may cause disease in the new fish or the
opposite may occur.

Sanitation
Good sanitation practices will help reduce the load of
potential pathogens. Such practices include proper system
maintenance by removing excess suspended particulates,
uneaten food, and dead or dying fish; reducing organics
and removing wastes.
Disinfection

Ongoing maintenance should include a disinfection

regime. Elimination of pathogens from nets, siphons and
tanks through the use of chemicals (such as chlorine,
benzalkonium chloride or iodine-containing compounds),
ultraviolet radiation, or ozone will help reduce the potential
spread of disease. Foot baths placed at key points leading
into and away from quarantine areas or areas with sick fish
will also help. Nets should be kept off the floor to avoid
contact with pathogens that may be harbored there.

System Disinfection or Sterilization

As described previously, water may spread

pathogens and also be a potential reservoir for them. Water
from a tank containing sick fish often carries numerous
disease-causing microorganisms. When this same water
enters another tank of fish, those fish are then exposed to
the microorganisms and they will have an increased risk of
developing disease. Disinfection helps to greatly reduce the
spread of some pathogens. Two techniques commonly used
to disinfect water in aquaculture systems are ultraviolet
sterilization and ozonation.

Ultraviolet Sterilization

Ultraviolet (UV) sterilizers typically consist of UV-

producing lamps encased in a glass or quartz sleeve. Water
is passed over the lamps. The lamps emit ultraviolet light (a
wavelength of approximately 254 nm is considered optimal)
that penetrates cells and damages genetic material (DNA
and RNA) and proteins.
 For each type of microorganism, a specific “zap
dose,” measured in microwatt seconds per square
centimeter, is required to selectively sterilize the system
(i.e., kill the unwanted organism). The zap dose is
determined by the intensity or wattage of the lamp, contact
time or flow rate of the water, water clarity, and size and
biological characteristics of the target organism. In general,
larger organisms require a larger zap dose; however, the
specific structure of certain viruses (which are generally
much smaller than bacteria) makes some of them more
difficult to “kill” than other larger organisms. In general, the
zap dose required is lowest for gram-negative bacteria, and
it increases progressively for gram-positive bacteria,
viruses, spore-forming bacteria, and protozoans.

Ozonation

Ozone-disinfection systems introduce ozone, O3, a

highly reactive molecule, into a contact chamber (isolated
from the main system holding the fish). Ozone generators
are more complex than UV-sterilizing units, and they
require the presence of a high energy field through which
dry filtered air or pure oxygen flows. The ozone oxidizes
(i.e., reacts with and breaks down) dissolved and suspended
molecules, as well as molecules within and on pathogens in
the water. In freshwater systems, ozone rapidly breaks
down or dissipates once it makes contact with the water;
therefore, water from a contact chamber can be
reintroduced into the system quickly, if the system is
designed properly. However, as ozone is so highly reactive,
all ozone must be eliminated from the water prior to its
reintroduction. This elimination can be accomplished in
several ways including off-gassing or removal by carbon
filtration. Ozone does not appreciably oxidize ammonia (i.e.,
convert ammonia into nitrite). In aquaculture systems, this
 reaction is commonly accomplished by nitrifying bacteria in
the biofilter. Ozone does oxidize nitrite to nitrate, so it
augments the efforts of nitrifying bacteria in the biofilter. If
the ozone is turned off in a system adapted to its presence,
the nitrifying bacteria in the biofilter may not be present in
high enough numbers to prevent nitrite levels from
temporarily spiking in the system.

In addition to sterilizing water, ozone helps other

parts of the system. Ozone promotes water clarity by
rapidly breaking down dissolved and particulate organics
that discolor or cloud the water. Ozone improves
biofiltration efficiency by decreasing the organic load in the
biofilter. This organic load is a food source for bacteria
known as heterotrophs. Heterotrophs include many
disease-causing bacteria that often compete with nitrifying
bacteria for space and oxygen in the biofilter.

Ozone is more dangerous than UV sterilization. Small

amounts in the water can kill fish and residual amounts in
the air can be toxic to humans. In seawater, removal or
dissipation of ozone is typically slower than in freshwater,
and by-products of ozonation can increase the risk of
disease in fish. For example, chemical by-products of
ozonation have been suggested as one potential cause of
head and lateral line erosion, although, research to
demonstrate this association has not be completed. Also,
some species are much more sensitive to residual ozone
levels than others. There are different ways to monitor
ozone levels in the water holding the fish. Therefore,
antibiotics should only be used in a bath under the
following conditions:

 the fish will not consume a medicated feed

 in consultation with a fish veterinarian or fish health
specialist
 proper culture and sensitivity tests have been run to
determine which antibiotic should be used
 the treated fish are “taken off line”, so the water is
contained and will not contact the biological filter
 the treated water is disposed of according to local
regulations

Use of Antibiotics and Antibacterials

Antibiotics are drugs of natural or synthetic origin

that have the capacity to kill or to inhibit the growth of
micro-organisms. Antibiotics that are sufficiently non-toxic
to the host are used as chemotherapeutic agents in the
treatment of infectious diseases of humans, animals and
plants. In aquaculture, antibiotics have been used mainly
for therapeutic purposes and as prophylactic agents.
Diagnosis of a bacterial disease should be verified by a fish
health specialist, and appropriate bacterial tests should be
run to determine which antibiotic will be effective. Ideally,
in ornamental fish, antibiotic should be given in a
medicated food, although in some cases injections may be
warranted for valuable individuals.

In the ornamental fish industry and in public aquaria,

antibiotic bath treatments are used, under strict control,
when fish are not taking food or have external infections.
Some of the antibiotics used in aquaculture are

Terramycin -- Terramycin is an antibiotic used to treat

systemic (internal) bacterial infections. It is approved by the
U. S. Food and Drug Administration (FDA) for the treatment
of sensitive bacteria of the genera Aeromonas,
Pseudomonas, and Hemophilus in salmonids and catfish. It
is used as a feed additive at a rate of 2.5 grams of drug
(active ingredient)/100 pounds of fish weight/day for 10
days. A 21-day withdrawal period is required before the fish
may be slaughtered and used for human consumption.

Sulfamerazine - Sulfamerazine is an antibiotic

compound for the treatment of furunculosis in salmonid
fishes. It is used as a feed additive at 10 grams of drug
(active ingredient)/100 pounds of fish weight/day for 14
days. A 21-day withdrawal period is required before the fish
may be slaughtered and used for human consumption.
(Note: Old fish health literature implies that Sulfamerazine
is an approved compound for use on food fish, this is no
longer true! Because many individuals were substituting a
generic "sulfa drug" for sulfamerazine, the manufacturer
decided to no longer manufacture Sulfamerazine, the
product for which FDA granted a label. Since Sulfamerazine
is no longer available, the use of any generic sulfa drug will
be known to be in violation of the law.)

Antibiotic used in aquaculture in India.

Antibiotic Purpose Mode of application

Oxytetracycline Myxobolus spp. Supplemented in the

feed

sulfadiazine+trimethoprim Ulcerative and Water dispersible

systemic type powder
(Aeromonas
hydrophila)

Chlor-tetracycline Not defined Supplemented in

feed

Romet-30 -- Romet-30 is a combination of two

antibacterial drugs that has FDA approval for the
treatment of furunculosis in salmonids and enteric
septicemia in channel catfish. It is used as a feed additive
in both cases at a rate of 50 milligrams drug (active
ingredient)/kilogram of fish weight/day for 5 days. A 42-
day withdrawal period is required for salmonids and a 3-
day withdrawal period is required for channel catfish before
the fish may be slaughtered and used for human
consumption.

In aquaculture, responsible conduct in the prudent use of

antibiotics should be to reduce their use to therapeutic
purposes alone; prophylactic use must be replaced by good
husbandry practices, including adequate hygiene
conditions and vaccination programmes. Among producers,
the priority should be education programmes that
emphasize proper drug use, e.g. which drugs are permitted
and how to comply with HACCP protocols. It should also be
noted that, for fish products, the use of dip solutions or ice
with antibiotics for the purpose of extending product shelf
life is forbidden.

Use of Other Chemicals

Research has shown that there are some differences in the

inhibitory effects of formalin, malachite green (illegal for use
in food fish), methylene blue (methylene blue is not
recommended by fish health specialists and is illegal for use
with food fish), copper sulfate, and potassium
permanganate on the biofilter bacteria.

Copper Sulfate -- Copper sulfate (CuSO4) is used to treat

a variety of external parasites of fish. It is also an effective
algicide and piscicide, it can kill aquatic plants and fish and
is approved for use as an algicide. The relationship between
toxicity of copper sulfate and alkalinity of the water is very
important. In water with an alkalinity less than 50
milligrams per liter (mg/L), copper sulfate can be toxic to
fish and it should not be used unless a bioassay has been
run in the water first with a limited number of the fish to be
treated

Formalin-F -- Formalin-F (formalin) is approved for use in

the treatment for a number of external parasites. It is
commonly used as an indefinite pond treatment at 15
milligrams per liter (mg/L). Formalin will remove 1 mg/L
dissolved oxygen for every 5 mg/L of formalin used as a
treatment. Therefore, if dissolved oxygen concentration in a
pond is low, aeration must be provided or a different
treatment should be used. Formalin must not be stored at
temperatures below 40 F because it will form very toxic
paraformaldehyde at low temperatures.

Potassium Permanganate -- Potassium permanganate

(KMnO4) is approved for use in aquaculture as an oxidizer
and detoxifier and has been used effectively against a
number of external disease organisms of fish. The normal
treatment is 2-8 milligrams per liter (mg/L), depending
upon the amount of organic matter in the pond to be
treated. Ideally, one would like to maintain a "wine red"
color in the water for a 12 hour period to ensure an effective
treatment. A preliminary bioassay can be performed with a
small volume of culture water to determine the appropriate
dose for the system. By contrast, potassium
permanganate experiments have been mixed. In one study,
a 4-mg/L dosage resulted in no inhibition of the biofilter,
whereas in another study, a 1-mg/L dosage resulted in an
86% inhibition.

Formalin used in one study, at 25 mg/L had no effect,

whereas another study showed reduction of biofilter
bacterial activity by 27% when used at 15 mg/L. As a rule
of thumb, most aquaculturists do not consider use of
formalin at 15–25 mg/L to have a major impact on the
biofilter. However, when testing for ammonia levels,
formalin will react with Nesslers Reagent (a component of
most ammonia test kits) and can give a falsely elevated
ammonia reading. In systems treated with formalin, the
salicylate reagent test for ammonia is recommended (Hach
Company 2002) because it does not react with aldehydes
(e.g., formaldehyde found in formalin).

Malachite green (again, illegal for use in food fish) has

been shown to have no effect on the biofilter at 0.1 mg/L,
combined with or without formalin at 25 mg/L. Copper
sulfate at 1 and 5 mg/L likewise had no effect on
biofiltration.

The actual impact on an individual system will most likely

depend upon many factors, such as chemical
concentration, length of time in treatment, organic load,
pH, temperature, alkalinity, filtration, oxygen levels, and
stocking density; and, although this will most likely be true
for most chemicals, this may better explain the differences
in effect by potassium permanganate.

Salt (sodium chloride)

Another chemical commonly used in aquaculture systems

is salt (sodium chloride). Salt can be useful in reducing
certain parasite infections in a system. Salt also helps
reduce osmolarity stresses by increasing the salt
concentration in the water relative to the normal
concentration in the fish's body (freshwater fish have a
higher body salt content than freshwater and so must use
energy to keep the body's salt concentration – the
osmolarity – in balance). Most tropical fish can tolerate a
salt concentration of 1–3 g/L, and this level is not harmful
to the biological filter.

Avoidance of Disease
Fish are aquatic animals, they live in the water.
Water provides the oxygen they breathe, the food they eat,
and the means to dispose of their wastes (e.g., carbon
dioxide, urine, feces). The quality of the water determines
how well the fish will grow and, indeed, if they will even
survive. Maintenance of suitable water quality greatly
reduces the likelihood of a disease problem. Critical water
quality parameters include temperature (particularly
sudden and dramatic shifts), dissolved oxygen, pH,
alkalinity, hardness, nitrogenous wastes (unionized
ammonia, NH3; nitrites, N02 -), and toxic substances (e.g.,
heavy metals, pesticides, carbon dioxide). Many of these
parameters are interrelated; for instance, as pH and
temperature increase the proportion of Total Ammonia
Nitrogen (TAN) in the toxic unionized form (NH3) increases.
Water quality should be monitored frequently and
corrective measures initiated if conditions become stressful
(e.g., dissolved oxygen below 3 ppm for warm water fishes
or 5 ppm for cold water fishes, unionized ammonia above
approximately 0.02 ppm, temperature above 210 C for cold
water fishes). Use of a high quality feed provides fish with
the nutrients that they need to remain healthy and to grow
rapidly. Fish fed a nutritionally complete diet are better able
to cope with stress and to resist disease. Culturists should
remember that even high quality feeds will deteriorate if
improperly stored or kept too long. Feed should be
purchased from a reputable supplier, stored in a cool and
dry place, and used within 90 days of manufacture. Light
(excessive or rapid changes in intensity), noise and other
disturbances can stress fish and should be minimized.
Routine maintenance, stocking, and harvest require that
fish be handled. When fish are removed and processed (e.g.,
weighed, transported) they compensate physiologically. To
reduce the trauma of handling make sure all necessary
materials (e.g., nets, hauling tanks, scales) and adequate
personnel are immediately available. Use of salt (O. 1-0.5%
by weight in the water), aeration or oxygenation, and
anesthetics can reduce the stress associated with handling.
Handle the fish gently and for as short a time as possible.
Responding to Disease Problems
Regardless of how careful one is, if culture fish is cultured
long enough one will inevitably encounter a disease
problem. When a disease problem develops, a quick and
effective response is essential. There is no better
preparation than to know fish. Under routine aquaculture
conditions, healthy fish display “normal” behavior. Fish
feed vigorously when food is presented or shortly thereafter.
In ponds and cages, fish are usually invisible, except when
feeding. Therefore, it is important for the aquaculturists to
note the feeding behavior of the fish being cultured even
when automatic feeders are used. A reduced feeding activity
should serve notice to the aquaculturists that immediate
further investigation for the cause is warranted. In
raceways fish normally swim leisurely, either en masse or
singly, depending on the species. Distribution in raceways
varies for species, but is usually constant (e.g., some
species prefer covered areas and others prefer uncovered
areas, some concentrate toward the water inflow and others
are more randomly distributed). As a culturist, one should
become familiar with the normal behavior of fish. If their
behavior changes (e.g., they stop feeding, swim near the
water surface, dart or scratch on objects), something has
occurred and you need to find out what. The first response
to a disease is abnormal behavior; to recognize what is
abnormal, one must first be familiar with what is normal.
 Routine monitoring of water quality in a production
system is imperative. When abnormal behavior is observed,
culturists should check their water quality (e.g., dissolved
oxygen, nitrogenous wastes, temperature). A sudden
change in weather (e.g., a cold front moves through) can
cause a change in behavior. If water quality is a problem
(e.g., low dissolved oxygen, high unionized ammonia) then
corrective measures should be initiated. If abnormal
behavior persists for several days or mortalities are
observed, culturists should seek professional assistance.
Water quality data should be provided to the diagnostic
laboratory as well as information concerning the fish in
question.

Once a diagnosis has been made, the diagnostic laboratory

will identify the disease as well as recommend an
appropriate and approved treatment or action. In certain
cases a change in management is necessary. In other cases
it is necessary to add an antibiotic to the feed (for internal
bacterial infections) or add a chemical to the water (usually
for external parasite infestations). It is extremely important
that the aquaculturist follow closely the recommendations
of the diagnostic laboratory and take appropriate
precautions prior to the application of any disease
treatment.

TREATMENT OF DISEASED FISH

Over the years four cardinal rules of fish disease

treatments have evolved:

a. Know your fish

b. Know your water

 c. Know your chemical

d. Know your disease

The culturist must know his/her fish. What is their

normal behavior, what conditions are likely to stress them,
and to what diseases are they most susceptible. Some
chemicals are safe and legal to use on certain species and
ages of fish, but they may not be appropriate or approved
for your fish.

The quality of water influences the condition of fish.

Each fish species has a preferred temperature. Some fish
are more tolerant than others of reduced oxygen, high
turbidity, and elevated levels of ammonia. Water chemistry
in some systems remains relatively uniform (e.g., single-
pass system, properly functioning recirculated system). In
other systems, such as ponds, water chemistry can vary
widely on a seasonal basis or even during a 24 hour period.
Dissolved oxygen and temperature may change
dramatically each day, but alkalinity and hardness vary
little in ponds. In a properly functioning aquaculture
system, dissolved oxygen and temperature remain relatively
constant throughout the day and growing season, but
alkalinity and hardness can change in a matter of days. In
an improperly functioning or overstocked system, dramatic
and rapid changes in dissolved oxygen, ammonia or nitrite
can result in high mortality rates of cultured fish.

One of the most important aspects of knowing the

chemical has become knowing which chemicals are
approved for use by aquaculturists. Each chemical can be
used to treat effectively and legally a few to several diseases.
No one chemical is appropriate for all diseases or
situations. For instance, an antibiotic can be very effective
in the treatment of a bacterial infection, but is useless if the
disease is caused by a protozoan parasite. All chemicals
have precautions and considerations associated with their
use. If an aquaculturist has no experience with a particular
chemical a small group of fish should be treated first, as a
test before the entire lot is treated, to avoid potentially
heavy losses due to toxicity associated with overtreatment.
Extreme caution should be practiced when applying any
chemical treatment. Water quality influences the toxicity of
certain chemicals and is adversely affected by some
chemicals. The culturist should be knowledgeable of the
water quality in the culture facility. Of particular interest
are dissolved oxygen, alkalinity, and the amount of organic
material in the water. Since copper sulfate is an algicide,
consideration must be given to dissolved oxygen in a pond
to be treated. If a pond already has low dissolved oxygen,
an alternate treatment should be used. Copper sulfate will
only aggravate low dissolved oxygen problems by killing the
primary source of oxygen (the algae) and by adding a large
biological oxygen demand in the form of dead and
decomposing algae.

CONCLUSIONS AND RECOMMENDATIONS

Septic conditions account for the majority of bacterial

diseases of fishes. For this reason, in fishes, the period from
infection with pathogens to their death is short and,
therefore, it is very difficult to treat fish diseases excepting
the case in which it is possible to diagnose a fish disease
early and treat it at its early stage. Pathogens of fish
diseases usually live in the water at a fish farm but their
pathogenicity is not potent, therefore, they do not cause
infection in healthy fish. But when fish become unhealthy
and more susceptible to pathogens, they are infected and
great damage is caused. As mentioned above, the majority
of pathogenic bacteria in fish diseases are facultative
pathogens.

To protect fish from various bacterial diseases, it is first

necessary to improve the technology for fish culture and to
culture healthy fish, which are highly resistant to diseases.
Secondly, early treatment methods for fish diseases, as well
as diagnostic procedures for early detection of fish diseases,
should be established. Thirdly, it is necessary to carry out
studies on artificial immunization with vaccine and prevent
diseases of cultured fishes.

INHIBITION OF DEVELOPMENT OF FISH DISEASES BY

IMPROVEMENT OF MEANS OF FISH CULTURE
TECHNIQUES

The most important objectives under this should be as

follows:

To make studies on optimum nutrition contained in feed for

cultured fishes and on the optimum amount of feed to be
given to cultured fishes.

To make studies on environmental conditions suitable for

cultured fishes and establish a feeding procedure which
does cause any trauma in cultured fishes.

To treat cultured fishes in a manner which does not cause

any trauma in fishes, since indigenous pathogens invade
the body of fish via such trauma.

To make studies on a method to culture fishes which are

healthy and highly resistant to pathogenic bacteria so as to
inhibit development of diseases.
ESTABLISHMENT OF EARLY DIAGNOSTIC
PROCEDURES

Studies are needed to speed up isolation and identification

of pathogenic bacteria. For this purpose, facilities where
various pathogenic bacteria are preserved and diagnostic
anti-sera are produced should be reinforced. In addition, a
serological diagnostic procedure and a diagnostic procedure
using the fluorescent antibody method should be
established. For this purpose, it is necessary to provide
complete facilities where these studies are made.

INHIBITION OF DEVELOPMENT OF FISH DISEASES BY

IMMUNIZATION VACCINES
Studies on mass immunization of cultured fishes for the
prevention of fish diseases should be facilitated. Mass
immunization of fishes may be performed with three types
of vaccines, i.e., oral vaccine, immersion vaccine (bath
immunization) and spray vaccine. On the basis of these
vaccines, studies to develop new immunization vaccines
suitable for various fish diseases should be made. Since the
effectiveness of immunization vaccines depends on the
kinds of fishes and the types of fish diseases, an
immunization vaccine should be studied for each kind of
fish and for each type of each disease.

USE OF PROBIOTICS
The research of probiotics for aquatic animals is increasing
with the demand for environment friendly aquaculture. The
use of probiotics or beneficial bacteria, which control
pathogens through a variety of mechanisms, is increasingly
viewed as an alternative to antibiotic treatment. The term
probiotics is generally used to denote bacteria that promote
the health of other organisms. An expert with the Joint
Food and Agriculture Organization of the United
Nations/World Health Organization (FAO/WHO), stated
that probiotics are live microorganisms, which when
consumed in adequate amounts, confer a health benefit for
the host. A number of preparations of bacterial mixture are
nowadays commercially available and have been introduced
to fish farming as feed additives, or are incorporated in live
fish feed in order to increase the performance. Typically, the
lactic acid bacteria (LAB) have been widely used and
researched for human and terrestrial animal purposes, and
LAB are also known to be present in the intestine of healthy
fish. Other commonly studied probiotics include the spore
forming Bacillus spp. and yeasts. Multiple ways exist in
which probiotics could be beneficial and these could act
either singly or in combination for a single probiotic. These
include: inhibition of a pathogen via production of
antagonistic compounds, competition for attachment sites,
competition for nutrients, alteration of enzymatic activity of
pathogens, immunostimulatory functions, and nutritional
benefits such as improving feed digestibility and feed
utilization. Some of the examples of commercial probiotics
used in aquaculture are GROVIMIN TM, GROWLIFE TM,
BIOLIFETM and PROLIQTM. .

Tejpal Dahiya, Ravi Kant Verma and V. P. Saini

 Importance Of Soil And Water In Fish
Health Management

It is well known that productivity of a fish pond depends

largely on the abundance of fish food organisms and also
on occurrence-of congenial environmental condition in the
pond. The first step in the food chain of a fish pond is
constituted by primary food organisms e.g. phytoplankton,
which derive their nutrients from the pond environment
and with the help of solar radiation undergo photo
synthetic activities. Occurrence of these nutrients in pond
water and maintenance of its relevant chemical condition
depend largely on the nature and properties of the bottom
soil wherein a series of chemical and biochemical reactions
continuously take place resulting in release of different
nutrients in overlying water and also their absorption in the
soil mass. Considering this importance of bottom soils in
maintaining productivity of fish ponds described such soils
as the "Chemical laboratory of the fish pond".

Characteristics of pond soil and water

The single major phenomenon which differentiates a

fish pond soil from an upland one is the presence of a layer
of standing water above the bottom soil. As a result of this,
oxygen supply to the bottom soil becomes restricted and a
reduced or anaerobic condition develops below a thin
oxidized layer in the bottom soil surface. Characteristics of
these two layers are entirely different. The oxidized or
aerobic layer in the soil-water inter-phase is characterized
by presence of oxidized forms of various nutrient elements
viz., nitrate (NO3, sulphate (SO4), ferric iron (Fe***) etc and
also aerobic group of micro-organ isms. The lower reduced
or anaerobic layer, on the other hand, exhibits reduced
forms of the elements viz. ammonium (NH4') sulphide (S~),
ferrous iron (Fe***) etc. and also anaerobic group of
microorganisms. The magnitude and dynamics of these
oxidation-reduction reactions play an important role in
governing most of the chemical, biochemical and microbial
behaviours in the pond soil and water and also in
maintaining congenial environmental conditions. Important
behaviours of some of these parameters influencing
productivity of fish culture systems have been discussed
here.

pH and relevant properties

The major changes associated with electro-chemical

properties of fish pond soil and water are reflected in pH
and also the electrical conductivity EQ values. The pH of
acidic soils usually tends to increase and that of an alkaline
soil tends to decline with increase in the period and extent
of submergence. In addition, pH values of water exhibit
marked diurnal variation especially in case of productive
ponds owing to release of oxygen and consumption of
carbon dioxide during photosynthetic activity of primary
food organisms in daytime and steady increase in
concentration of carbon dioxide during night hours. Ne to
slightly alkaline pH ranges for both soil and water are
considered to be congenial aquatic production owing to
greater availability of most of the nutrient elements and due
to increased biological activities under this pH range.

Under acidic conditions of fish ponds, liming is

widely practised to raise the pH of pond and water. On the
other hand, under extremely alkaline situation, gypsum
may be required to reduce the pH to a congenial level.
However, application of lime or gypsum above lower the
required amount may be either ineffective or even harmful
to productivity ponds. In addition to studies on pH of pond
soil and water therefore, methodologies determining lime
requirement (GR) appear to be important.

Soluble salts and associated parameters

Submergence tends to increase the solubility of

many elements in the soil phase and thus helps to increase
electrical conductivity (EC) of pond soil and water. Under
freshwater conditions these changes do not usually reach
very high values so as to influence the productivity through
effects of total soluble salts. However, this behaviour may
encourage more solubility of some particular ions in some
cases and thus may alter ionic equilibrium of the system.

Like EC salinity also refers to amount of soluble salts

in water and is of special significance in case of brackish
water aquaculture where highly saline tidal water is used
for culture of fishes and prawns and variation in salt
content influences the productivity of the culture

Primary nutrient elements

Among different nutrient elements, nitrogen (N),

phosphorus (P) and potassium (K) are considered to be
primary nutrients owing to their major roles in plant
nutrition (Dev, 1995). Of them, P is generally considered to
be the most critical single factor in the maintenance of pond
fertility owing to its utmost importance to primary fish food
organism and also occurrence of very low amount of this
element in available form. This problem of availability of P
in fish pond soil and water is due to fixation of phosphate
ions into insoluble compounds of calcium (Ca) under
alkaline environments and iron (Fe) and aluminium (Al)
under acidic conditions resulting in release of very low
amount of P in available form under either situation.

Nitrogen (N), the second primary nutrient element in

fish pond nutrition, becomes available to primary fish food
organisms in two inorganic forms viz. ammonium (NH 4*)
and

nitrate (NO3). N in the bottom soil remains mostly in organic

form and, there from, is gradually mineralized to inorganic
(NH4) and (NO3) forms in which N becomes available to
primary producers in fish pond soil and water.

Inspite of forming the third primary nutrient element

influencing pond productivity, little work have so far been
done to understand the chemistry of potassium (K) in fish
ponds. High clay and organic matter content of productive
fish pond soils, associated with alkaline pH values usually
tend to maintain moderate to good amount of K in pond soil
and water. K becomes available to fish food organisms in
water soluble form in water and in water soluble as well as
exchangeable form in soil.

Presence of base elements

In addition to N, P and K, three other elements viz.

calcium (Ca), magnesium (Mg) and sulphur (S) constitute
the forum of major nutrient elements to be released
primarily by the soils and are called as secondary nutrients.
Apart from providing nutrition to fish food organisms, the
first two elements along with sodium (Na) and also K serve
as important components of the pond soil and contribute to
maintenance of alkaline pH in the pond environment.

Total concentrations of all divalent cations is total

hardness of water and is expressed in terms of ppm of
calcium carbonate (CaCO3) equivalent. Since Ca and Mg are
the two major divalent cations in most of the fish ponds;
total hardness also provides an assessment of occurrence of
these two base elements in water.

Major anions in fish ponds

Chloride (Cl), sulphate (SO4=), bicarbonate (HCO3)

and carbonate (CO3=) constitute the major anions in fish
pond ecosystem, in addition, hydroxyl (OH-) ion occasionally
registers its presence in appreciable amount, especially
under highly alkaline conditions. CO3- and HCO3- ions
provide CO2 in water which is required for photosynthesis.
These two ions and also OR form the major sources of
alkalinity and can be estimated through determination of
this parameter. In the soil phase occurrence of CO 3 and
HCO3- in the form of calcite, dolomite, etc can be assessed
from the amount of alkaline earth carbonate which is also
expressed as free calcium carbonate. In addition
watersoluble forms of these anions and also Cl - and SO4=
extract. Apart from being an important nutrient element, Cl -
ion remains associated with majority of soluble salts in soil
and water and hence estimation of this ion helps to assess
the total quantum of soluble salts to a fairly accurate
degree. Sulphate is a major source of sulphur (S), an
important constituent of protein However, under highly
reduced condition SO may be converted to sulphide (S=)
form and affect productivity of the ecosystem.

Organic compounds in pond soil and water

A wide range of organic substances occur in pond

water. Such materials include compound s like sugars,
starches, amino acids, peptides, proteins, humic acids etc
concentrations and relative distribution of which depend an
sever-al factors. These dissolved organic matters release
considerable amount of nutrient: elements to soluble form
and thus aid to fertility status of ponds. High
concentrations of organic matter, on the other hand, may
consume large amount of oxygen from the pond water
during its course of mineralization and thus create oxygen
tension in the aquatic environment. If this demand for
oxygen becomes very high, anaerobic condition may develop
in the poor especially in the bottom, resulting in
development of unhygienic In view of these effects of
organic compounds on pond environment, the methods of
detan of dissolved organic matter content of pond water
appears to be important- In addition. determination of
chemical oxygen demand (COD) of water, which indicates
tatally amount of oxygen required for oxidation of all the
organic matter in a water sample provides a measurement
of organic matter in water- I the rate of oxygen consumption
by and bacteria in the water sample within a specific time
and condition is expressed Marien ica oxygen demand
(BOD) which also generates an idea of organic load in band
is, a compound which often occurs in considerable amount
in organically rich vown nad requires to be estimated under
various environmental impact assessment programmes.
Although not confined to organically polluted waters
always, concentrations of free chlorine (Cl,) sometimes need
to be tested as a component of water pollution.

Fish pond sods, in general, exhibit comparatively

higher amount of organic matter (OM) than corresponding
upland soils. Organic matter plays an important role in
determining productivity of fish ponds. Apart from
influencing various physics properties of the bottom soils
alongwith releasing different nutrient elements to available
form in pond environment, it also controls the oxidation-
reduction reactions in the ecosystem. However, all the
components of organic matter in soil are not equally active
and, thereby, useful for productivity of soils.

WATER PARAMETERS

Physical Temperature

pH

Colour

Transparency

Chemical Conductivity

TDS

Salinity

Dissolved Oxygen (0)

 Carbon Dioxide (CO)

Hydrogen Sulphate (H,S)

Alkalinity / Acidity

Hardness

Chloride, Sulphate

Ammonia

Ammonium

Nitrite

Nitrate

Phosphorus

Heavy Metals

BOD

Soil and water chemistry plays a vital role in

freshwater aquaculture, such as selection of site, Pond
manuring, Management of water quality and Feeding
Management etc.

Soil parameters:

Physical parameters Soil Texture

pH

ORP and
Conductivity

Inorganic salts Soluble carbonates,

Bicarbonates
 Chloride and Sulphate

Exchangeable Ca, Mg, Na,

and K

Organic matter Organic carbon %

Nutrients Available phosphorus

Available nitrogen and

Available potassium

pH of the soil Soil can be classified on the basis of pH

Soil pH Type of Soil

4.0-5.0 Highly acidic

5.0-6.5 Moderately acidic

6.5-7.5 Near neutral

7.5-8.5 Mildly alkaline

8.5-9.5 Highly alkaline

Optimum range for aquaculture – 6.5 to 7.5

Adverse condition less than 4.0 and more than

10.0

Indication:
1. If pH goes out of optimum range, then, phosphorus
deposits in the form of insoluble aluminum phosphate and
calcium phosphate. As such, it is not available as a
nutrient

2. Less availability of NH,-N.

3. Effects of nitrogenous fertilizers also depend on soil

pH.

4. Effect on primary productivity.

5. Microbiological activities are also effected.

6. Decomposition of organic maters.

7. Acidic and alkaline soils are more prone to fungal

and bacterial load, respectively.

8. Extreme values of pH adversely effect metabolism of

culturable species.

Soil Texture:

Soil texture refers to the relative proportion of

primary particles of sand, silt and clay and other skeletal
materials in the soil body. Silt clay ratio is an important
criterion.

Optimum Range

Soil Properties Range

Clay(%) 30%±5%

Silt (%) 30%±5%

Sand (%) 40%±5%

Physical properties determined

1. Water retention

2. Absorption & release of nutrients

3. Organic matter

4. Availability of minerals

Doses of Lime to maintain optimum range of pH

Soil pH Lime (kg/ha) CaCO3

4.0-4.5 1000

4.5-5.5 700

5.5-6.5 500

6.5-7.5 200

8.5-9.5 Nil

Merits of Lime application:

1. It corrects the pH of soil and establishes a strong pH

buffer system, this system comprises CO,-HCO 3, -
CaCO3, preventing wide fluctuations in pH.

2. The presence of calcium in lime, speeds up the

decomposition of organic matters, releasing carbon
dioxide from the bottom sediments.

3. Lime, by its toxic and caustic action, kills bacteria as

well as parasites in their various life history stages.

OXIDATION-REDUCTION POTENTIAL (REDOX

POTENTIAL)
 It is a vital parameter in controlling the biological
treatment of sediments. Redox potential should be high for
better aerobic treatment (+ 200 mV to + 6000 mV) and low
for better anaerobic treatment (-100 mV to -200 mV).
However, much reduced values are not desirable.

CONDUCTIVITY

Conductivity of the soil is directly related to the total

excliangeable dissolved solids & salinity. Conductivity of
soil in Punjab has been observed to range between (dSm -1)
0.05 to 0.88.

INORGANIC SALTS

Inorganic soluble salts like Carbonates,

bicarbonates, chloride and sulphate reach out to the water
and play an effective role in determination of water
alkalinity, hardness, pH and salinity.

On the other hand, exchangeable Calcium and

Magnesium determine hardness of while Potassium
inclicatesthe nutrient availability.

ORGANIC MATTER (Organic carbon)

Extent of organic carbon contained in soil depends

on soil texture. Observed na. organic carbon range (Punjab)
0.00 1 % to 0.915%, Optimum range 0.5 % to 1.5%.

Organic compounds in soil are more complex and

varied than the mineral constituents degree of bacterial
activity depends both on the carbon content and the
carbon/nitrc (C/N) ratio. Their activity becomes less when
the ratio is below 10 and improves where 20 or above.

NUTRIENTS - Optimum range for prawn culture

Available phosphorus 3-6mg/100g soil

Available nitrogen 25-50 mg/1OOg soil

Available potassium 1-2%

The standard combination of N:P:K as 18:8:4 is

commonly used to fertilize the pond, dose of which is
decided on the high, medium and low criteria of the soil.

Available P2O5 Available N

(mg/100g soil) (mg/100g soil)

High 6-12 50-75

Medium 3-6 25-50

Low 3 25

Water parameters

TEMPERATURE

Water temperature depends on the atmospheric

temperature and depth of water. If dept is less than a
meter, temperature shoots up water temperature can be
maintained b increasing the volume of water and by regular
exchange/addition of fresh bore well water Optimum
temperature range for prawn ponds is 26 to 31ºC. It plays
an important role r biological activity of the pond.

pH

pH of water is an important factor regulating the

growth and survival of prawns. Many of the chemical and
biological activities in ponds are related to pH value. A pH
value of 7.0 - 8.5 is optimum for the growth of prawns. pH
above 11.0 and below 5.0 retards growth. Acidic pH can be
controlled by limiting and alkaline pH can be controlled by
applying gypsum.

COLOUR & TRANSPARENCY

Colour and visibility of pond water gives a visual

assessment regarding nutrient status (natural productivity)
of pond ecosystem. The colour of pond water maybe due to
the presence of humic acid, fulic acid, metallic ions as iron
and magnese suspended matter, phytoplankton and
weeds.. Light brown to light green is good for growth of
prawns. Visibility depends on the light intensity and light
penetration in the water colour. A properly maintained
pond will have visibility upto 20 to 35 cm: If the value
decreases, it indicates increase in nutrient but if the value
increases, it indicates decrease in nutrient.

CONDUCTIVITY, TDS, SALINITY

Conductivity is the measure of capacity of water to

conduct electric current. It depends on total dissolved
solids. Salinity of water can be measured by direct
multiplication of electric conductivity with multiplying
factor. Range of salinity should be below 7.0 ppt for
freshwater aquaculture.

DISSOLVED OXYGEN

Dissolved oxygen is one of the most important

parameters in water quality assessment and reflects the
physical and biological process prevailing in water. It
depends on temperature, pH and total biomass of pond.
Diffusion of oxygen in pond takes place from atmospheric
air and photosynthesis. During day time, plants in the
water supply planty of oxygen into the water by
photosynthesis and during night time; they utilize oxygen
from water for respiration. So, there is diurnal fluctuation
in the level of dissolved oxygen in the water. The minimum
level is observed during early morning and maximum level
is observed during afternoon hours.

Optimum dissolved oxygen level required in ponds is

5.0 to 10.0 ppm. Dissolved oxygen below 3.0 ppm is lethal
for prawns. Oxygen depletion can be controlled by aeration
and regular water exchange.

CARBON DIOXIDE (CO2)

Dissolved CO, of the water sample should not be

more than 3 ppm. In natural water, pH also changes
diurnally and seasonally due to variation of photo synthetic
activity, which increases the pH due to consumption of CO,
in the process. Most of the alkalinity in natural water is
formed due to dissolution of CO in the water.

HYDROGEN SULPHIDE (H2S)

Hydrogen Sulphide has some toxic effect on the

organism present in the water. It is h corrosive in nature
and promotes growth of certain bacteria. It is found in
higher concentration when orgains matter is high and redox
potential is low reducing condition microorganisms use
sulphate as electron acceptor to decompose organic matter
convert pollution. US Environmental Protection Agency has
put a limit of 0.05mg/l.

ALKALINITY/ACIDITY

Alkalinity/Acidity of the water is its capacity to

neutralize a strong acid/base and characterized by
presence of ion. In natural waw, it is due to free hydroxi
ions a hydrolysis of solid formed by weak acids and strong
bases.

Total alkalinity = HCO3- + 2CO3-2 + OH- H+

Alkalinity is also produced by action of water on

limestone or chalk.

CaCO3 + H2O + CV Ca (HCO3)2

(insoluble) (Soluble)

Total alkalinity represented the productivity of the

pond. Optimum range for prawn culture depends on types
of alkalinity.

30 to 50 ppm asC03-2

50 to 250 ppm as HC03-

HARDNESS

Hardness is the property of water, which prevents

the lather formation with the soap an increasing the boiling
point of water. Principle cations imparting hardness are
Calcium an Magnesium. However, other cations such as
strontium, iron and magnese also contribute to the
hardness.

The anions responsible for hardness are mainly

bicarbonates. carbonates, sulphatrz chloride, nitrate and
silicate etc. Hardness is of two types : Temporary and
Permanent Temporary hardness is due to bicarbonate salts
and easily removable. However, permanenhardness of the
water is not easily removable. It is due to carbonate,
sulphate, chloride nitrate and silicate salts.
 Optimum range for hardness of pond water is 50 to
150 ppm. If the range is below 30ppm, then plankton does
not grow.

CHLORIDES AND SULPHATES

Chlorides and sulphates play an important role to

increase hardness of the water and level of salinity. In ease
of saline water, it is an important parameter for sites
selection in prawn culture.

Ammonia (NH3-N)/Ammonium (NH4-N)

Ammonia of mineral origin is rare in natural water.

The most important source of ammonia is the
ammonification of organic matter. Occurrence of ammonia
in water can be accepted as the chemical evidence of
organic pollution. Ammonia in higher concentration is
harmful to fish/prawn and other biota. Toxicity of ammonia
increase with pH because at higher pH, most of the
ammonia remains in the gaseous form. The decrease in pH
decreases its toxicity due to conversion of ammonia into
ammonium ion, which is much less toxic than the gaseous
form. Permissible limit of ammonia nitrogen (NH,-N) is less
than 0.1 ppm. However, incaseof NH4 - N, less than
1.5ppm.

Nitrate (NO,'- N) & Nitrite (NO,'- N)

Nitrate ions have no mineral source in natural

waters. Nitrite represents an intermediate form during de-
nitrification and nitrification reaction in nitrogen. It is very
unstable and gets converted into. either ammonia or nitrate
depending upon the conditions prevailing in the water. The
permissible limit of nitrite nitrogen is less than 0.1 ppm.
 Nitrate represents the highest oxidized form of
nitrogen. The most important source of nitrate is biological
oxidation of organic nitrogenous substances produced
indigenously in water. Atmospheric nitrogen fixed into
nitrates by the nitrogen fixing organisms. The high amounts
of nitrate denote the aerobic conditions and high stability of
the wastes. The permissible limit of nitrate nitrogen is less
than 1.0 ppm.

PHOSPHORUS

Phosphorus in the natural freshwater is present

mostly in inorganic form such as H 2PO4- HPO4-2 and P04-3.
The phosphorus content of natural freshwater is low and
biological growth is limited due to its insolubility in water.
The optimum range of phosphorus is 0.3 to 0.5 ppm.

HEAVY METALS

Heavy metals are those having a density of more

than 5 times higher than that of water. They are usually in
trace amounts in natural waters but many of them are toxic
even at very low concentration. Some of these heavy metals
get biomagnified in the waters and ge-. accumulated in
higher trophic levels e.g. fish, crabs, plankton, molluscs
and other aquatic organisms. Sites having heavy metal
contamination should be avoided for aquaculture.

BIOLOGICAL OXYGEN DEMAND (BOD)

BOD is the amount of oxygen utilized by

microorganisms in stabilising the organic matter On an
average basis, the demand of oxygen is proportional to the
amount of organic waste to be degraded aerobically. Higher
range of BOD value indicates the increase in growth of
bacteria in the system. Hence, chances of 'disease
occurrence increase, as well. The permissible of BOD is less
than 30 ppm.

Tejpal Dahiya, Ravi Kant Verma and V.P. Saini

An Introduction To Bacterial Fish Diseases

Aquaculture has been established as an industry in India

in the last decade. Indian major carps that are Catla catla,
Labeo rohita and Cirrhinus mrigala are cultured on large
scale particularly in Andhra Pradesh, West Bengal,
Maharashtra, Tamil Nadu, Andman-Nicobar Island, Punjab
and Haryana. In 1994-95 bacterial fish disease alone
caused lose of more than 3500 crores of rupees but today it
is estimated around 6000 crores of rupees.

To sustain the Aquaculture industry appropriate

health management involving development of diagnostic
methods and treatments are necessary because about 7.5%
of total annual income of the country is comes from
aquaculture. Today rapid and sensitive diagnostic
biochemical and DNA based methods are used. The
following bacterial fish diseases, their causative organisms,
symptoms and treatments are discussed below:

1. Dropsy
2. Tuberculosis
3. Bacterial gill disease
 4. Bacterial kidney disease
5. Pop eye disease
6. Edwardsiella septicemia / Edwardsiellosis
7. Bacterial hemorrhagic septicemia
8. Vibriosis
9. Columnaris
10.Hemophilosis
11.Tail & fin rot
12.Furunculosis
1. Dropsy
Causative Organisms: - Aeromonas hydrophila and
Myxosporidian sp. which is a parasite.
Symptoms: - (a) Diseases fish are characterized by
abnormal accumulation of fluid inside the body
cavities. The fluid is of straw colored which gives a
swelling appearance to the fish.
(b) Due to abnormal accumulation of fluid the
scales protrude out ward direction.
(c) Superficial ulceration all around the body in
epidermis is seen.
(d) Hemorrhages on body surface.
(e) Exophthalmia:- It is bulging of eyes outside.
(f) Yellowing of gills and lot of mucus is found
between the gills.
Susceptible fish species: -
1. Catla 2. Rohu 3. Mrigala and 4. Common
Carp
Reporting time: - It is reported during Feb-
March & October-November at intermediate
temperature.
Treatments: - Terramycin is very effective when
given along with feed for at least two weeks.
2. Pop Eye Disease
This disease is primarily reported in Ornamental
fishes but also in cultured fishes all over the country
 Causative bacterial organism: - Flavobacterium sp.

Symptoms: -
(a) Emaciation: The body of the fish is of pale
color due to loss of scales all around the
body.
(b) Multifocal white nodules are found in
internal organs especially in visceral
organ, retina, Choroid and brain.
(c) Granulomas which are the clusters of
bacterial colonies are found in all visceral
organs and retina, choroids and brain of
the diseased fish. In this disease fish
suffers from opacity and ultimately due to
blindness it is died.
Treatments: - It is treated by bath in KMnO 4 at the
concentration of 2mg /liter of water for the period
of one month. Various types of broad spectrum
antibiotics which are effective against gram
negative bacteria are used to cure these diseases all
over the country.
3. Bacterial Gill Disease
It is found all over the country. Fry are the most
susceptible to this disease but all the ages may be
affected from BGD.
Causative Organisms: -
Flexibacter columnaris
F. psychrophilus
Cytophaga psychrophilus
Flavobacterium sp.
Symptoms: -
1. Necrosis of gill lamellae: The gill lamellae of the gills
are narcotizes first at the distal end which further
travels towards the proximal end of the gill lamellae.
 2. Suffocation and surfacing: - Due to necrosis of gill
lamellae the fish is deprived of O 2 due to which it
suffocates and comes ate the surface of water to
inspire more O2 i.e. Surfacing.
3. Hyperplasia of gills.
The predisposing factors for this disease are
overcrowding, accumulation of metabolites waste
products particularly NH3, organic matter in water
and increase in temperature of the pond.
Treatments:-
It is treated by dip bath method. Bathing in KMnO 4 for
one week continuously at the concentration of
2mg/litre. It is also treated by bath using antibiotics
drugs like chloramine for a short duration at the
concentration of 10ml/litre.
4. Tuberculosis

It is also called as Mycobacteriosis. All marine, brackish,

freshwater and aquarium fish species are affected from this
disease.

Causative organisms:-

Mycobacterium narium.

M. chelonei.

M. fortituitum.

Symptoms:-

1. Anoxia:-It is a problem of water when water

quality is bad and very less oxygen is dissolved in water due
to which fish deprives of oxygen and it starts surfacing

2. Emaciation: it is loss of scales from the body

surface due to which fish look pale in color.
 3. Eyes are bulging towards the outside i.e.
exophthalmia

4. Vertebral deformities.

5. Loss of normal colorations of the body which is

changes to pale color.

6. When fish is dissected and histopathologically

studied under microscope lots of granulomas are seen
which contains numerous bacteria in them. These
granulomas are seen in all internal organs of the fish.

7. Source of initial infection of this disease is

ingestion of bacterial contaminated feed or debris.

8. Once aquarium is infected it is very difficult to

eradicate this disease except depopulation of the aquarium.

Treatments:-

1. It is treated by using broad spectrum

antibiotic drugs which are very effective against all types of
gram negative bacteria.

2. Bath in tetracycline for a long time also cure

this disease. It can be also cured by using tetracycline in
feed.

3. Teramycin Sulphate at the rate of 10mg /100

gm of body weight along with feed for 4-7 days is given to
cure or control this disease.

5. Bacterial Kidney Disease

It is reported in all fish species but it is a serious disease of

salmonids and brook trout.
Causative bacterial organism: It is caused by
Renibacterium salmonarium.

Symptoms:

1. Eyes of the fish are bulged towards the outside

i.e. Exophthalmia.

2. Skin coloration changes to dark i.e. skin

darkening. Vesicles and ulcers develop on the skin.

3. Congested blood vessels are seen at the base of

the skin which gives red appearance to the fish it is also
called as hemorrhages.

4. Abscissions, cavitations and contraction of

muscles are reported.

5. Swelling of kidney is reported.

6. Spleen have numerous white nodules i.e.

granulomas which are found in parenchyma. These
granulomas have gram positive bacteria init. These bacteria
are also reported from their internal organs like liver,
kidney, heart and other internal organs of the fish.

Treatments: - It is treated by using antibiotics which are

effective against gram positive bacteria. Bath in maracyn at
the rate of one tablet for 37.8 liters of water.

Transmission: It is transmitted by direct contact with

contaminated fish. Bacteria enter through epidermis and
then it becomes a systemic disease.

6. Columnaris
It is also called as “Cotton wool disease”. It is found in both
cold and warm water fishes like salmonids, catfish and
Indian major carps. It is reported in warm summer months.

Causative bacterial organism:

Flexibacter columnaris

Cytophaga columnaris

Symptoms:

1. Lesions occur at head, back and gill regions.

As disease progresses lesions spread.

2. Lesions on gills first appear on distal end and

then moves towards proximal end.

3. Whole gill suffers from necrosis.

4. It is mainly reported at higher temperature 25-

35°C when overcrowding, injuries and poor water quality.

Treatments:

It is treated by bath treatment and giving

antibiotics along with the feed. Bath CuS04 for 20 minutes
at the concentration of 33mg/litre. Bath in KMnO4 at the
rate of 2mg/litre for prolonged period . Incorporation of
oxytetracycline in feed at the rate of 0.1gm/kg of fish feed
or at 3% of body weight along with bath treatment.

7. Edwardsiellosis

It is also called as Edwardsiella Septicemia. It primarily

reported in channel catfish but also observed in goldfish,
golden shrines, large mouth bass and the bull head. It
affects eels seriously.
Causative organism: Edwardseilla tarda

Symptoms:

1. Skin coloration is changed by having patches of

red color.

2. Hemorrhages in skin and muscles of the body.

3. Red colored patches develops into lesions which

filled by gas and forms cavities which are called as
malodorous.

4. In this disease fish looses control over the

posterior half of their body and continue to eat.

Treatments: Lodophor has been found to be very effective

but water quality improvement in the hatchery is the most
essential component for keeping this disease away.

8. Bacterial hemorrhagic septicemia

It is found all over the country. It occurs at high

temperature when overcrowding.

Causative organisms:

Aeromonas hydrophila

Pseudomonas fluorescens

Symptoms:

1. Hemorrhages in skin, fins, oral cavity and

muscles. Epidermis is superficial ulcerated.

2. Exophthalmia
 3. In spleen, liver, kidney and heart multi focal areas
of necrosis with numerous rod shaped bacteria are
reported.

Treatments:

It is treated with oxytetracycline at the rate of 50-

75mg /kg of body weight for 10-20 days.

Cefax 2 litre for one hectare with 40kg CaCO3 in two

equal doses with two days difference can cure this disease.

Transmission: It is transmitted via contaminated water or

diseased fish.

9. Furuculosis

It is also called as “ulcerative disease of goldfish”. It

occurs mainly in salmonids but it is also reported in cold
and warm water fishes. It is reported during summer and
early rain season.

Causative organism:

It is caused by Aeromonas salmonicida.

Symptoms:

Subcutaneous swelling which causes ulcerative

dermatitis, hemorrhages in muscles are seen. Large deep
ulcers are seen in the skin of the fishes. Bacteria multiply
in the blood vessels and capillaries and destruct the wall of
blood capillaries.

Treatments:

It is treated with using terramycin at the rate of

0.07g/kg of body weight along with feed. It is also treated
with sulphamerazineat the rate of 0.25g/kg of fish body
weight daily for 10-15 days consecutively.

10. Vibriosis

Vibriosis is also called as “red boil disease”. It occurs

predominantly in marine water and brackish water but it
occasionally reported in freshwater also.

Causative organisms:

Vibrio alginolyticus

V.anguillarum

V. salmoncida

Symptoms:

It is characterized by hemorrhages in skin of the tail, fins

and muscles. Ulcers in skin are reported. Anatomically,
necrosis of liver, kidney and gut mucosa is seen,
hemorrhages in entire body cavity are reported.

Treatments:

1. It is treated with suphamerazine at the rate of

0.25g/kg of fish body weight per day and after two days
interval repetition of the same dose.

2. Furazolidone at the concentration of

50-70g/kgof fish body weight for a period of 10 days.

11. Hemophilosis
 It affects freshwater teleosts.
Causative organisms:

-Aeromonas hydrophila

-Haemophilus piscium

Symptoms:

1. It is characterized by open sores on the body

surface.

2. First white spots appears on body surface

which enlarge to form circular shallow dark red or grey
abscess in fins , edges of jaws and roof of the mouth .

3. Internal organs like spleen gets necrotized and

reduced.

4. Oedema of dermis is also reported.

Treatments:-

It is treated by using oxytetracycline at the rate

of 27-75 mg /kg of fish body weight per day for 35-45 days.

12. Tail and fin rot

It is the erosion and disintegrations of fins tails. It is

reported all over the country in cultured fishes. It mainly
occurs before monsoon period.

Causative organisms:

Aeromonas hydrophila

Pseudomonas fluorescens

Symptoms:
 1. It is mainly characterized by erosion and
disintegration of fins and tails of the fishes. Initially
formation of white area on the margins of fins is noticed
which further extends at the base of the fins. Outer margins
become frayed and disintegration of soft tissue between fin
rays starts and continues till total fin is lost.

2. Epithelial hyperplasia is reported.

Treatments:

It is treated by using different types of antibiotics

and chemicals at different doses

1. It is treated by Terramycin at the rate of 10-

15 mg / kg body weight of the fish which
mixed with feed.
2. Bath in chloroamphinicol solution of
concentration 60mg/litre at temperature 23-
27°C for 6 days (Mathew, 1961).
3. Dip bath treatment in chemical mixture of
59ml formalin, 59ml of malachite green and
113 liters of water.
4. Bath in CuSO4 at the concentration of
0.5g/litre for 1-2 minutes.

Ravi Kant Verma, Tejpal Dahiya and V. P. Saini

 Diseases Of Catfish And Their Control

As fish production has become more intensive, the

potential for serious disease outbreaks to occur has also
increased. Diseases of cultured catfish have presented
management problems. Since a couple of years it has been
seen and estimated that about 50% losses in our catfish
culture area due to infectious diseases. It is essential that a
catfish farmer recognizes clinical signs of both infectious
and non-infectious diseases in order to prevent and control
the same.

Three factors must occur simultaneously or

concurrently for an infectious disease outbreak to develop
(1) The presence of a pathogenic organism (2) Susceptible
fish and (3) A pre-disposing (stressful) condition.
Experienced fish farmers take precautions to prevent the
simultaneous occurrence of these three factors through a
programme of good management. In general these
precautions constitute the preventive medicine or health
maintenance programme of a fish hatchery or farm. Fish
that are fed well, under crowded and in a good environment
are less likely to develop a disease.

Bacteria are among the most important

pathogens of catfish which may cause extensive losses of
the cultured catfish populations. Mortality due to bacterial
pathogens are often associated with environmental stresses,
because, the causative agents are usually saprophytic,
facultative or opportunistic and occur naturally in most
waters. Pond water with high levels of material is good
many potential bacterial pathogens.

BACTERIAL DISEASES

A variety of bacteria produce infectious

diseases in fish and the diseases occur frequently in
nursery rearing and grow-out ponds caus serious concern
to aquaculturists. A large number of pathogenic bacteria
like Aeromonas hydrophila. Flextbacter columnaris.
Edwardstella tardc Pseudornonas fluoresceris. Streptococcus
and staphylococcus sps etc - are responsible for causing
variety of disease eg., columnarls disease, tai and fin rot,
bacterial speticaemia, tropical ulcerative disease, eye
disease, etc. The control of infectious diseases of fish
remains the single most important factor in the success of
aquaculture an realization of this fact is increasing day by
day among the farmers.

AERMONIASIS

The motile aeromonads have attracted much

attention enteropathogens and a number of strains have
been shown to produce enterotoxic substances. The name,
motile aeromonad disease, has been accepted for disease
previously referred as haemorrhagic septicaermahydrophilu
and A.. punctata have been described as primary secondary
pathogens of fishes throughout the world. The disease oco
as an acute, subacute, chronic or latent disease of fishes.

External sings of the disease include erythema

(redness)at the of the base of the fins in and around the
mouth, within the opercula and around the anus. The
oroanisms are also capable of production severe skin
lesions to ulceration of various

Internal signs are characterized by severe

congestion and petechial hemorrhages in the peritoneum
and most of the viscera organs. Intestinal mucous Is
usually severely congested and contain bloody i-nucus arid
fluid. Slicing through the muscle may show pinpoint
hemorrhages. The fins arc usually frayed with hemorrhage
at the base.

The body cavity may be filled with bloody fluid

and kidney may be soft arid swollen. Liver if often pale with
minute hemorrhages. Lower intestine arid vent are
frequently hemorrhagic and swollen and mucosa filled with
bloody or yellowish mucus.

Diagnosis of the disease is usually made on

the basis of necropsy findings arid through isolation and
identification of bacteria. Aerorrionads are gram negative
coccobacilli which can be demonstrated in squash
preparations from various body organs. There are no
specific antiscra which can be used for rapid identification
due to the enormous strain variations. However, definitive
diagnosis can be accomplished by biochemical activity of
the organism.

Mortality as high as 80% may occur in

physically stressed, nutritionally deficient, young fish.
Older fish may show more resistance pathogens can be
isolated from Morbidity may however reach 100% and the
intestinal tract arid kidney of all fishes.

Fish can be protected from aeromonas

infection by reducing physical stress, nutritional
deficiencies and injuries leading, to epizootics. Fishes or
fish eggs with aeromonads should not be transported from
infected geographical areas to non-infected areas.

TREATMENT AND CONTROL

Oxytactracycline-50-70 body weight in feed for 10 days.

Sulfamerazine-200 mg/kg body weight for 7 days in the

feed.

Eternal disinfectants may be used once a week in the water

to reduce the population of bacteria eg. Methylene blue@ 4
mg/ 1.

ULCERATIVE DISEASE

A less virulent strain of A. hydrophila is responsible

for the disease which is characterized by distrinct
ulcerations of the opaercula and head region. In severe
cases, ulcerations are seen deep in the musculature and
cranial bone. Moralities maybe encountered in these cases.

CONTROL

Drug sensitivity test should be done before advocating

suitable medicine for controlling the disease.

Topical application of potassium permanganate extremely

effective.

COLUMNARIS DISEASE
 This disease is caused by a bacteria known as
Flexibacter coluinnaris which afl-CcLs fish of all ages. In
per acute cases the animals my die without showing any
gross pathological lesion, which is usually due to a
virulent strain of pathogen. In virulent strain the lesiol are
characterized by white spots on the body of fish, often on
the head, lips or ions arc mostly covered with flakes
disease is often known as "cotton wool disease". Gradually
the lesiolis raised hemorrha,,ic zone appear on the
inflamed skin. Localized lesions may also appear in the
gills. Often due to presence of large number of spread to
(whole body, and grayisli white prollf-crating epithelium
clangle over the body. After 3-4 days small circular areas
surrounded by raised hemorrhagic zone appear on the
inflamed skin. Localized lesions may also appear in the
gills. Often due to pathogens the colour of the lesion may
turn orange or yellow. Potassium permanganate
bath at a concentration of 1:50, 000 for 15 minutes or
direct application of potassium permanganate over the
body lesions for 2-3 minutes are very effective methods for
controlling columnarls disease.

CONTROL

Copper sulphate at a concentration of 1:2,000 for 15

minutes followed by a bath with running water is also
recommended.

Chloramphenical or chlortetracycline bath at 10-20 ppm

are found to be effective.

HAEMORRHAGIC SEPTICAEMIA

The etiology of hemorrhagic septicaemia is still

under dispute. The disease is clinically etiology of disease
is clinically indistinguishable from acromonas infection,
although Pseudonionas fluorescens -has often been isolated
from diseased fish showing septicaemic lesions

The disease is manifested two forms i.e. either

moralities. In acute cases, severe congestion and
hemorrhagic are noticed in the visceral organs, wlicrcas in
chronic cases tile disease is characterized by fibrinous
peritonitis. Petechial hemorrhages are seen in the Internal
well of the air bladder.

Oral administration of oxytetracycline or

intraperitoneal Inject- of kanamycin have been reported to
give good results.

VIBRIOSIS

This is one of the most significant disease of

cultured fish usually caused by Vibn'o anguillurum.

Other species which infect both freshwater as

well as saline water fish are V. parahaemolyticus and V.
alginolyticus. Diseased fish show haemorrhages in the
mouth regi opercula and ventral surface of the body.
Hyperaemic intestine, swollen spleen and necrotic kidney
are other gross pathological findings. Periorbital and
abdominal dropsy may also develop. Congested and pulpy
kidney are other gross pathological findings.

CONTROL

Prevention of the disease can be achieved by

immunization. However, the disease can be controlled by
feeding a diet containing 0.02% furazolidine for two weeks.
Sulphamerizine @, 80-120 body, height is recommended as
a suitable control measure.

BACTERIAL GILL DISEASE

 Bacterial disease is to a combination of
unfavourab) condition with infeculon of- gills by
myxobacterla of' the genus cyLophaga. Besides this, a large
numb(.-r of-rarn negative bacteria have been reported in gill
lesions.

The disease is characterized by proliferation of

gill epithelium and in extreme cases fusion and necrosis of
the gill filaments. The affected fish show sluggish movement
and signs of asphyxiation and surfacing. Poor
environmental condition and over crowing are the main
triggering factors.

CONTROL

Dimethyl benzyl ammonium, chloride at 2 ppm for I hr

(bath treatment) or Ethyl mercuric phosphate at the same
dose have been used successfully to control the disease.

EDWARDSIELLOSIS

The causative organism is Edwardslella tarda, which

produces extreme emaciation, anaemia, loss of skin, pceling
off and dropping of skin and gas filled abscesses with foul
smelling, in fish. The disease was, termed as
"emphysematous. putrefactive discase" aptly describing the
gross appearance of infected fish. Edwardsiella has been
ii-riplicated in jastroenteritis and meningitis.

Factor which-can prevent the disease. lodine

preparation in diluted f'orm can cure the disease if applied
for 3-4 days.

FIN ROT AND TAIL ROT

Mostly the fry and fingerlings are affected with this

disease, although older ones are not spared. Several
pathogens such as Aeromonas liquifaclens, A. formicans and
Pseudomonas fluorescens have been attributed to the
typical fin rot and tail rot of fish which is characterized by
progressive erosion and disintegration of fins and tail. The
first sign is whitening of the outer margin of the fin which
progresses towards the base. The continuous degenerative
process terminates at the complete loss of the fin and/or
tail.

The best method of prevention of the disease is to

maintain favourable environmental condition and proper
sanitation. Over crowding is an important factor which has
got a major role to play in these conditions.

CONTROL

Dip treatment in a I i I mixture of formalin and mala~hite

green

Dip treatment in 1:2000 copper sulphate for I to 2 minutes,

are very effective, Besides this

Calclurn hypochloritc (lip at 2 ppm for 2 minutes and

treatment of water with tetracvdine @100 mg/gallon for- 5
days help in curing the disease completely.

EPIZOOTIC ULCERTATIVE SYNDROME (EUS)

This disease is widely prevalent among catfishes In

Our Country- Being one of the primary victim of EUS, all
species of catfislics exhibit typical ulcerative Icsions
throughout the body. The posterior third of the body
including the tall, brim of he mouth and lateral
musculature in C. butrahcus and H. fossilis show severe
haemorrhagic: and necrotic lesions which are often
secondarily Infected with bacteria and /or fungi. The
diagnosis is based on the clinical signs like lethargy,
listlessness, ariorexia arid gross ulcertaive lesion. Although
there is difference of opinion about the primary causative
agent of EUS, it is emphatically proven at Cl-FA that A.
hydrophilu plays the salient role outbreaks of the epizootic.

Although the disease can be moderately controlled by

application of lime but best wav to control EUS in catfishes
is by application of CIFAX ( A medicine developed by CIFA).
A single dose @ 500 ml/ha-m water area is enough to
control the disease within 3-7 days. Application of a
mixture of turmeric powder and lime has also been
advocated in the control of the disease.

FUNGAL DISEASES

Fungi are important disease causing organisms of

catfish and their eggs. Generally fungi are secondary
invaders to other disease & mechanical injury. Spores of
waterborne fungi attach to the injured which are known are
Lissues of' the fish and establish a color mycelium. Once
the colonies are established they proliferate to invade the
healthy and eventually causes death office animal.

There, are several species of fungi cg. Saprolegnia,

Achlya Brachiomyces, Mucor, Penicilium try infection "'I
catfishes. Among these saprolegnia is the most commonly
encountered genera in catfish diseases. The rate of
mortality among infected fish is high, therefore the best
control of saprolegniasis is by prevention or health
maintenance – fungi -Sanitation and hygienic culture
practices are essential for control of fungal diseases in
catfish. 1'-2 min dip in 2-3% salt solution may be useful. A
mixture of malachite green (0.05 ppm.) and formaldehyde
(20 ppm) proves beneficial in most of the fungal diseases.

PROTOZOAN DISEASES
 Protozoan diseases are among the most significant of
all parasitic diseases in catfish. General management
condition, stocking density, acre and size of the f-ish may
influence the susceptibility.

CONSTIASIS

This is caused by a small, rounded kidney-shaped

flagellate known as Ichthyobodo which attach to the gills
and external surface of the fish causing some grayish white
erosions. Ichth obodo is a tiny protozoar. about a size of a
red blood cell and can be seen easily under the their
microscope. Formalin (15-25ppm). Potassium
permanganate (2-3 ppm) or cooper be used as a prolonged
treatment copper sultphate (0.5-1 ppm) may be used as
prolonged treatment of lchtyhbodo.

ICHTHYOPHTHIRISIS

'Ich' caused by I. multifilis is one of the most

important parasite which can cause lot of damage in the
catfish culture system. 'Ich, is climate proto/mill parasite
characterized by relatively large size and C shaped nucleus
in adult parasite. The parasite burrows into the skill and
Causes white pin-head size spots which can be seen with
naked Cyc. Ich epizootics are temperature related and are
most prevalent at 2528 (1C. The disease is difficult to
control. But experienced fish farmers have controlled it
either by use of Kmn04, formalin or Copper sulfate by
frequent treatment (every alternate clay).

OTHER PROBLEMS UNRELATED CRACK-HEAD

DISEASE

TO INFECTIOUS AGENTS
 Besides infectious agents, there are sOrne other
riaortatity problems in catfishes especially in C.
batrachuswh ich has often baffled the catfish cultists in our
country as well as in some African and South-East Asian
countries. Large scale moralities have been observed in C.
batrachus with big hemorrhagic cracks at the base of the
skull resembling dentition of hack-saw. No causative
organism could be isolated from these affected fish which
baffled the scientists. However, is "butted to Vitamin C
deficiency in the diet by some workers from Nigeria, who
could control the malady by supplementing adequate
amount of Vitamin C in the diet.

Tejpal Dahiya, Ravi Kant Verma and R.C. Sihag

 An Introduction To Fresh Water Prawn
Diseases

It is believed that fresh water prawn (scampi),

Macrobrachium rosenbergii is less susceptible to diseases
than penaeid shrimp (Panaeus monodon). It might be as a
result of lower stocking densities used in culture practices
and less transfer of brood stock. However, a variety of
diseases have been reported in larval, juvenile and adult
scampi, which include fouling protozoans such as
Zoothamnium, Vorticella and Epistylis; fungal pathogens
such as Lagenidium, Sirolpidium and Fusarium; bacteria
such as Vibrio, Aeromonas, Leucothrix etc., and also non-
pathogenic diseases such as stress and neurotic symptoms,
moult failure and so on. This may be due to shorter life
cycle and faster metabolism of these animals, which results
in weaker resistance to pathogens. Prawns are highly
sensitive to pathogens during molting period when they
cannot feed, have a week body and poor mobility. The
incidence of disease is higher in low quality water,
particularly if oxygen levels are low. Prawns need good
quality water and enough plant life. The following diseases
of fresh water prawn were reported by various workers:
1. Larval Midcycle Disease

It is widely occurs in Thailand, Philippines, Malaysia and

India. The larval stages IV to XI are usually found to be
affected from this disease. The symptoms appeared about
10 days post-hatching and high larval mortality was
observed. The weak larvae stop feeding and settle at the
bottom of the rearing tank and exhibit spiral swimming
behaviour along with vertical axis turning. The larvae
become bluish grey in colour and the hepatopancreatic
epithelium get atrophied

Possible causes

This disease might be due to nutritional deficiency,

since lecithin deficiency is often said to occur along with
this disease. Bacteria, Enterobacter aerogens has been
reported to be associated with this disease.

Preventive measures

Infected stock should be immediately destroyed and

tanks should be disinfected and dried. The farmers should
adopt standard hatchery techniques that may prevent
vertical or horizontal transmission of this disease.

2. Bacterial diseases
a) Bacterial necrosis

This is a common disease observed in larvae, post-

larvae or adults in fresh water prawns. It is variously
termed as 'black spot', 'brown spot', 'burnt spot', 'Shell
Disease' or Chitinolytic bacterial disease. This is caused by
the invasion of chitinolytic bacteria, which break down the
chitin of the exoskeleton, leading to erosion and
melanization (dark brown to black pigmentation) at the site
of infection. Several chitinolytic bacteria (Gram- ve rods)
such as Vibrio spp., Pseudomonas spp., Aeromonas spp.,
Spirillium spp. etc are involved. This disease reduces the
value of the harvested prawns, apart from causing
mortalities.

This disease can be controlled in captive and cultured

populations by reducing over crowding, proper husbandry
and system hygiene.

b) Filamentous bacterial disease

Filamentous bacteria such as Leucothrix muco,

Thriothrix spp., Flexibacter spp. etc. sometimes cause
mortalities in fresh water prawns by discolouration of gills
and associated secondary infections. The larvae or the post
larvae become moribund, with reduced mortality, poor
feeding and growth. Increasing husbandry and hygiene
standards will improve the situation.

c) Luminescent bacterial disease

Luminescent bacteria especially Vibrio harvey often

infect the prawn larvae reared in hatchery conditions, when
they appear in seawater. They spread very fast and poor
hatchery conditions increase their virulence. The infected
larvae become fluorescent in dark light, with reduced
feeding and in severe cases mass mortalities occur.

Treatment

This disease can be cured by using specific

antibiotics and disinfecting the pond water

3. Fungus Disease
 Larval mycosis due to Fusarium spp., Lagendium
spp., Sirolpidium spp. cause severe mortalities to prawn
larvae often as a secondary infection. F. solani, F.
moniliformes, L. callinectes etc. are commonly found.
Fungus infects the dead or damaged tissue caused by
wounds or other infections resulting in locomotory
difficulties due to mycelial growth. In serious infections of
Lagendium extensive non-septate highly branched mycelia
invade through out the body replacing all the tissues.
Specialized hyphae protrude through the cuticle.

4. Viral Disease
Scampi is believed to be free of viral diseases a case of
occurrence of white spot virus has been reported from
Taiwan in M. rosenbergii. Signs closely resembling white
spot syndrome have been observed on its exoskeleton as
reported by Peng et al. (1998).

5. Protozoan parasites
Fouling protozoans such as Zoothamnium, Vorticella etc. are
of common occurrence in larval, post larval or adult
scampi. They attach to the body and appendages and
disrupt mobility and feeding. Mortality occurs only in severe
cases of susceptibility. Formalin treatment will be helpful to
save the affected.

6. Other Diseases
a) White muscle disease

Opacity or 'white muscle' is a condition to the prawn

arising out of any stress such as over crowding, extreme
temperature or pH, low dissolved oxygen etc. The body
appears opaque white proceeding forward from the tail
region. The condition is often reversible once the stressed
condition is corrected.
b) Blackening of gills
 Blackening of gills of scampi may sometimes be found
as a manifestation of several other disease syndromes,
precipitation of dissolved chemicals, turbidity, vitamin C
deficiency etc. General discolouration of gills may occur due
to melanization of tissue and necrosis, which may be visible
through the side of carapace.

Vijayanti Jakhar and Sihag, R.C.

 Prominent Catfish Diseases

Catfish is a common name for about 2200 species of fishes

out of which around 1200 species are from South America.
The name was taken from the feelers or barbells that extend
from each side of the upper jaw of the fish (in some species
from the lower jaw also) resembling the whiskers of a cat.
These are usually freshwater fish without scales and having
long whiskers (barbells) around its mouth that are sensitive
to touch, taste and smell in nocturnal scavengers who live
near the bottom in shallow waters. The dorsal and pectoral
fins are often edged with sharp spines which may be
poisonous in some cases and used in defense causing
severe wounds. Members of several catfish families in South
America are covered with bony plates. A European species-
the sheatfish or Wels is the largest catfish reaching up to a
weight of 290 kg and a length of almost 4 m (about 13 ft).

Scientific classification:

Catfishes belong to the order Siluriformes of which two

families (Ariidae including sea catfishes Arius sp. and
Plotosidae including eel tail catfishes) are primarily marine
rest all other families inhabit freshwater. The freshwater
catfish families Siluriidae (sheatfish) consists of Silurus
glanis and Wallago and Clariidae includes walking catfishes
(Clarias batrachus –Indian catfish and Clarias garipienus –
African catfish). Bullhead (Ictalurus punctatus - the channel
catfish and Ictalurus furcatus is known as blue catfish)
belong to the family Ictaluridae.

Clarias batrachus –Indian catfish, Heteropneustes fossilis,

Clarias fuscus (Chinese catfish) are important cultivable
species of freshwater air breathing teleost catfishes. From
all cultivable catfishes Clarias macrocephalus is considered
to be more valuable due to having more tender meat but C.
batrachus grows faster and more easily available at the fry
stage as well as its market is higher than the other. In India
C. batrachus is grown on large scale in almost every state.
In the wild, clariids are mostly carnivorous feeds mainly on
small invertebrates like worms, aquatic insects (Anisops
and Corixa), crustaceans-small prawns (decapods) and
trash fishes (Ambassis renga, A. nama, Amblypharyngodon
mola, Esomus denricus and Punctius sp.) and also eat dead
vertebrates and other fishes i.e. scavengers. But in culture
in these are fed mainly on trash marine fish, supplemented
with rice bran, boiled broken rice or manufactured pellets.

Catfishes have high prices due to fewer bones and

also used for sport. They currently account for about one-
half of the aquaculture production in the world and
production is increasing every year due to the use of new
technologies like genetic improvement, recirculating
systems and control of diseases.

Factors causing disease

Aquaculture has transformed into an industry due to

increased food demand because of increasing human
population at a very high rate. To gain maximum economic
benefits intensive aquaculture practices are carried out
which alter hydrobiological parameters and lead to
reduction in stress which may be physiological or physical
and is primary contributing factor for fish disease and
mortality in aquaculture (Walters and Plumb1980). Many
potential fish pathogens are always present in the water,
soil mud and air cause fish diseases. Fish reared under
commercial aquaculture conditions and production units
are weakened by the following stress conditions:-

1. Increased fish density

2. Poor water quality (i.e. low dissolved oxygen,
undesirable temperature or pH, increased levels of
carbon dioxide, ammonia, nitrite, hydrogen sulfide,
organic matter in the water);
3. Injury during handling (i.e. capture, sorting and
shipping);
4. Inadequate nutrition and
5. Poor sanitation.

These conditions lead to stress on the fishes resulting in

decreased resistance of the fish a leading to the spread of
disease and parasitic infestation.

Signs of disease

1. Irregular swimming and loss of balance

2. Surfacing and gulping of air
3. Stop feeding
4. Fish become passive and does not show movements
Pathogenesis and causes of infection
Stress and injury initially trigger an alarm reaction of fight
or flight response which results the following series of
changes within the fish body:-

1. Stress stimulates endocrine glands to release

hormones which metabolize liver glycogen and
increase blood sugar concentration. This increased
blood glucose/sugar produces a burst of energy
which prepares the animal for emergency situation.
2. Hormones released from the endocrine glands
mainly adrenal gland also suppress the
inflammatory response (a defense used by fish
against invading disease organisms).
3. Disruption of osmoregulation (water balance) due
to changes in the metabolism of minerals. With the
results freshwater fish absorb excessive amounts of
water from the environment (over-hydrate) and
saltwater fish lose water to the environment
(dehydrate) lead to increased energy requirements
for osmoregulation.
4. Increase in respiration
5. Increase in blood pressure and
6. Releasing of Reserve red blood cells into the blood
stream.

Fish adapt themselves against stress for a short period and

may look and act normal. However, energy reserves
depletion and hormone imbalance suppress their immune
system and increasing their susceptibility to infectious
diseases.

Classification of disease

Diseases in catfishes are classified as Infectious and Non-

infectious.
 a. Infectious diseases may be endo or ecto - parasitic
(bacterial, fungal, viral, algal and protozoan, aquatic
insects- lernea, argulus, trematodes-). Infectious
diseases are spreads vary rapidly and becomes
endemic in a very short period of infection.
b. Non-infectious diseases are mainly genetic and
nutritional deficiency disease. Nutritional deficiency
disease occur only when the fish feed given to the
fishes is lacking any component like protein i.e.
essential amino acids- Histidine, Isoleucine, Leucine,
Lysine, Methionine, Phenylalanine, Valine,Threonine,
Tryptophan, vitamins, lipid and minerals etc.
Deficiency diseases can be cured by giving required
amounts of nutrients in the feed. Genetic diseases
occur due to the mutation or by the action of
teratogens.

In this chapter only few prominent diseases of catfishes are

discussed especially their causative agents/ pathogens,
symptoms, transmission, diagnosis and treatments.

Vibriosis

Vibriosis is also called “red boil disease”. It is a systemic disease

occurring mainly in marine fishes but occasionally in brackish
and freshwater. It is a major cause of fish mortality and has
been reported to inflict serious losses to the fisheries industry.
Although vibriosis is distributed worldwide, it caused severe loss
among cultured yellowtail in Korea. The incidence of vibriosis in
the fish farms is a matter of great concern for inland saline water
fisheries in India.

Pathogens
Serious epizootics caused by V. anguillarum, V. ordalii, or V.
salmonicida occur in pink salmon (Oncorhynchus gorbuscha),
chum salmon (O. keta), Atlantic salmon (Salmo salar), Japanese
eel (Anguilla japonica), yellowtail (Seriola quinqueradiata) and ayu
(Plecoglossus altivelis). V. anguillarum caused septicemia in
channel catfish (Ictalurus punctatus) and vibriosis in marine
fishes cultured in Japan.

V. anguillarum and V. alginolyticus are regarded as the dominant

species causing vibriosis, several other Vibrio species are also
pathogenic and have been found in finfish, shellfish and marine
sediments. V. vulnificus is usually encountered as a highly
virulent but opportunistic human pathogen, though infects eels
and causes development of red patches on the trunk or tail.
Although the seven Vibrio species reported as fish pathogens
may infect many marine and estuarine fishes, cultured fishes
are most susceptible due to stress (Dahiya, 2005).

Symptoms:-

Vibriosis is characterized by

1. Skin hemorrhages
2. Ulcers or septicemia
3. Anemia and arrhythmia
4. Red necrotic or boil like lesions in the musculature, on
the fin bases and mouth
5. Patches on the body surface
6. Hemorrhages in the gills and the viscera and
7. Inflamed intestinal tract
It is characterized by superficial ulcerations on all over the body
surface. As the infection progresses, the ulcers deepen up to
skeleton; leaving hemorrhages on the skin. The wounds are
characterized by swollen reddish boundary with grayish pink
coloration at the centre

I. Early stage: This is characterized by white spots and

superficial ulcers (fig. 1).

II. Mid stage: This is characterized by hemorrhages in

fins and skin (fig. 2).

III. Late stage: In this stage, the ulcers deepen up to

skeleton. The wounds are characterized by swollen
reddish boundary (fig. 3).
These symptoms are typical in vibriosis disease caused by
bacteria of the genus Vibrio and are similar to several earlier
reports on wide variety of fishes from other parts of the globe.
Incidence of A. hydrophila is common in nutrient enriched and
polluted fresh as well as brackish waters. This bacterium has
been reported to be pathogenic to human beings and causes
septicemia, wound and ocular, respiratory, bone and intra-
abdominal infections. Its entry in the saline water fish ponds
may be through cattle dung which is a common fertilizer for fish
ponds (Lakshmanaperumalsamy et al., 2005).

Diagnosis

1. Serological identification tests are mainly used for V.

anguillarum, V. ordalli, and V. salmonicida. and fish
pathogenic vibrios also differentiated biochemically.

Treatments

1. CEFAX at the rate of one litre per ha and the same

dose should be repeated after 4 days of first
treatment.
2. POVIDINE @ 1L/Hectare-meter and repeat the dose
after 4-6 days of first treatment.
3. HERBODINE @ 1L/Hectare-meter.
 4. It can be treated with sulphamerazine @ 0.25 g/kg of
fish body weight per day and after two days the same
dose is repeated.
5. Furazolidone at the concentration of 50-70 g /kg of
fish body weight for 10 days.
Prevention

1. Add water or maintain water level at 3 feet.

2. Immunization of fishes by using dead or alive
microorganisms or different kinds of treated antigens
from microorganisms. For administration of vaccines
different methods (injection, immersion, orally by
feed and spray) may be used depending upon fish
age, fish value either for marketing or may be used
as brooder fish as well as technical knowledge of
farmer or fisheries officer (Boon and Huisman, 1996).
3. Development of bacterial resistance and
immunosuppression by giving good quality and
quantity of feed as it is health determining factor like
vitamin C and dietary lipids.
4. Giving immunomodulator to the fish for potentiating
the immune system like glucan.
5. A general treatment can also be practiced to avoid
disease outbreak during all period of fish farming is
application of 15-20 kg chuna/lime (CaO), 1-2 kg
NaCl plus 100-200 gms of KMnO 4 for every 15th day
or two time in a month.
EPIZOOTIC ULCERATIVE SYNDROME

Introduction

Epizootic ulcerative syndrome (EUS) is caused by the

infection of commonly called fungi – the water mould from
class- Oomycete, Order-Saprolegniales, genera
Aphanomyces. EUS is also known as red spot disease
(RSD), mycotic granulomatosis (MG) and ulcerative mycosis
(UM). Recently, scientists proposed that EUS should be re-
named as epizootic granulomatous aphanomycosis or EGA.
At present, RSD, MG, UM and EGA are synonyms for EUS.

EUS was first reported in farmed freshwater ayu

(Plecoglossus altivelis) in Oita Prefecture, Kyushu Island,
Japan in 1971, later reported in estuarine fish grey mullet
(Mugil cephalus) in eastern Australia in 1972 from then
outbreaks were reportd from Papua New Guinea into
South-East and South and West Asia, reached to Pakistan.
EUS causes disease and mortality in farmed and wild fishes
over 50 species of fish have been confirmed by histological
diagnosis to be naturally affected by EUS out these
catfishes are walking catfish (Clarius batrachus and C.
gariepinus) and I. punctatus and wells (Siluridae) (OIE
2006).

Causative agents

The oomycete that causes EUS is known as Aphanomyces

invadans. Parasites and rhabdoviruses have also been
associated with particular outbreaks and secondary gram-
negative bacteria (A. hydrophila, A. punctata,
Flavobacterium sp., Psuedomonas sp., E. tarda, and Vibrio
parahaemolyticus) are isolated from diseased fish and
invariably infect EUS lesions (OIE 2006, Das 1997).
Dhanraj et al., 2008 isolated and identified 17 isolates from
the liver, gills, intestine and muscle of diseased C. striatus,
among these 17 isolates, dominant bacterial and fungal
species are Aeromonas hydrophila, Flavobacterium sp.
Aspergillus flavus, and Aphanomyces invadans. The
following ware the most frequently isolated bacterial and
mycotic flora with a prevalence of greater than 10%:
Aeromonas hydrophila, Aeromonas sp., Flavobacterium sp.,
Vibrio vulnificus, Staphylococcus sp., Yersinia enterocolotica,
Shigella sp., A. salmonicida, Aspergillus flavus, Aspergillus
sp. and Aphanomyces invadans. From 25 microbial colonies
in gill samples, 11 colonies of Aeromonas hydrophila were
identified predominantly.

Etiology and clinical characters

It has a complex infectious etiology and clinically

characterized by

1. Presence of invasive Aphanomyces infection and

necrotising ulcerative lesions leading to a
granulomatous condition.
2. Outbreaks occur mostly after heavy rainfall or during
low temperatures period which favour sporulation of
Aphanomyces invadans and low temperatures also
delay inflammatory response of fish against oomycete
infection.
3. Normally outbreaks occur in wild/weed fish first and
then spread into cultured fishes in to the ponds
(Saha and Pal 2002).
The symptoms may be same and specific in each disease
but sometimes few symptoms may be or seem to be
common as described in every disease.

Transmission

1. The zoospores of Aphanomyces transmit horizontally

from one fish to another through water supply. These
(zoospores) have whip like motile flagella and capable
 of attaching to the damaged skin of fish and
germinating into hyphae.
2. If zoospores cannot/unable to find susceptible fish
species or encounter adverse conditions then they
form secondary zoospores, later on secondary
zoospores encyst in the water or pond environment
and wait for favorable environmental conditions for
the activation of the spores.
3. It is still unclear that how the Aphanomyces
pathogen or its spore survives after the outbreaks
usually occur about the same time every year in
endemic areas.

Symptoms of Clinically affected fish:

The early signs of the disease include the following:-

1. loss of appetite

2. Darkening of skin.

3. Infected fish may float below the surface of the water

4. Hyperactivity with a very jerky pattern of movement.

Fish usually develop red spots or small to large

ulcerative lesions on the body. Red spots may be
observed on the body surface, head, operculum or
caudal peduncle (Ayyappan 2006). Large red or grey
shallow ulcers with brown necrosis are observed in later
stages. Large superficial lesions occur on the flank or
dorsum (Fig).
The lesions are more extensive and can lead to complete
erosion of the posterior part of the body, or to necrosis of
both soft and hard tissues that are exposed in the living
animal. fig

Diagnosis

Diagnosis is based on clinical signs and histopathology

 1. Early lesions of EUS are erythematous dermatitis
with no oomycete involvement
2. isolation of Aphanomyces invadans from internal
tissues as well as from skin and presence of mycotic
granulomas in histological section or
3. Non-septate hyphae of Aphanomyces invadans (12-
25 µm in diameter) can be observed microscopically
in muscle squash preparations of the infected area
around the lesion. Lesion scrapes normally show
secondary fungal, bacterial and/or parasitic
infections.
4. Strong inflammatory response and granulomas
around the penetrating hyphae.
5. using molecular diagnostic methods for identification
of Aphanomyces invadans in a very small time with
accuracy (Miles et al., 2003)

Control and prevention

Probably control of EUS in natural waters is impossible

when outbreaks occur in small, closed water-bodies.
Selection of disease free and resistant species for culture
purposes currently seems to be the most effective means of
farm-level control. Eradicate or exclude the fungus through:

1. Use of prophylactically-treated, pathogen free

hatchery reared fry
2. Use of well-water
3. salt bath treatments
4. Drying and disinfection of contaminated nets and
equipment.
 5. Removal of infected and wild fish,

Treatments

1. CIFAX at the rate of 1 litre per hectare-meter is

applied and marked improvement of the ulcerative
condition is seen within 7 days of application and
ulcers heal with in 10-14 days.
2. Liming of ponds at the rate of 250 kg per hectare-
meter prior to stocking as well as during outbreaks
as it improves water quality effective in reducing
mortalities and controlling the disease.
3. Neem along with branches and leaves is found to be
very effective. Neem paste can be made by the farmer
himself with composition of Neem leaves 12gms,
turmeric 3gms, glycerin 5ml and distilled water 5 ml
(Islam, 2003).
4. Aloe paste (aloe gel 10g, glycerin 5ml and distilled
water 5 ml) is also found very effective against
saprolegniasis (Hamiffa et al, 2006).

Saprolegniosis

It is also called as “winter kill syndrome” and is the single

largest cause of economic losses in aquaculture and fungal
infections are second only to bacterial diseases in economic
importance. Fungal infections are generally restricted to
chronic, steady losses. Hatai and Hoshiai (1994) indicate that
in Miyagi Prefecture, Japan, there is an annual mortality rate
of 50% in coho salmon (Oncorhynchus kisutch Walbaum) due
to Saprolegnia parasitica Coker. Fifty percent per year losses
have also been reported in elver (Anguilla anguilla) culture in
Japan and in the southeastern United States, major financial
loses occur in channel catfish farming due to a condition
called "winter kill." Some catfish farmers have reported losses
of up to 50%, an economic loss of $40 million (Bruno and
Wood, 1994).

It is present every where in salt and fresh water, cold, cool

and also warm environments. The spores undergo perenation
and very difficult to eradicate from the infected fish pond.
Both Saprolegnia and Aphanomyces belong to family
Oomycetes and white and tuft looking with cellulose in their
cell wall instead of fungal chitin. Both prefer to grow and
reproduced on dead fishes.

Causative organisms

Many fungi cause diseases that can infect and kill catfish
eggs, fry, fingerlings and adults and most fungal infections
are caused by water molds of the family Saprolegniaceae.
Within the Saprolegniaceae family, Saprolegnia sp.
(Saprolegnia parasitica, S. declina, S. turfosa, S. paradoxa,
S. australis and S. monoica), Achyla sp. and Branchiomyces
sp. are the genera that cause most disease in catfish. These
are fungal opportunistic facultative parasite either ecrophs
or saprotrophs, although some fungal species within this
family are primary pathogens (e.g., Saprolegnia parasitica),
most (e.g., S. declina and S. saprolytica) cause disease only
when there is preexisting illness, mechanical injury, or
environmental stress. Moreover, Saprolegnia may be
secondary invader to bacterial infection or parasitic agents.
However, the importance of saprolegnia as a primary
pathogen is still debatable where some outbreaks with
mass mortalities may occur in the absence of other
pathogens (Ramaiah, 2006, Duboon et al, 2006;
Bangyeekhun et al. 2001; Doriga and Martinez. 1998;
Goodwin 2002 and Li 1996).

Saprolegnia sp. grows well on egg chemo-tactically i.e. it

grow on egg shell/debris after hatching, when grow well it
moves toward new living egg. So, during breeding extremely
low survival of the resultant eggs and very low yield of
hatchling due to protracted spawning activity of the fish
damaged the developing eggs in the limited breeding space.
It is reported to be harmful in two ways:

a. The exudates released during the mating acts

pollute the ambient water to an extent that it
starts stinking and
b. The released eggs get fatally injured by the
movement of spawners. It may be mentioned
here that the eggs of C. batrachus are highly
adhesive by nature and if they are dislodged
from their fixed spot, their perivitelline
membrane gets ruptured.

The asexual spore of Saprolegnia release motile, primary

zoospores. Primary zoospores are active only for a few
minutes before they encyst, germinate, and release a
secondary zoospore. Secondary zoospores are more motile
for a longer period of time than primary zoospores and are
considered the main dispersion phase of Saprolegnia. The
repeated cycles of encystment and release, called
"polyplanetism", allow secondary zoospores to make several
attempts to locate a suitable substrate. Secondary
zoospores are considered the infectious spore of
Saprolegnia.

Following encystment, secondary zoospores release hairs

for attachment. It has been suggested that these hairs are
also used for buoyancy, to decrease the sedimentation rate,
and for fungal-host recognition response. The most
pathogenic species of Saprolegnia, S. parasitica, have long,
hooked hairs.

Different species of Saprolegnia are able to germinate under

different environmental conditions and nutrient levels.
Some Saprolegnia isolates are able to grow in water alone
and on waste products from hatcheries. S. parasitica can
sprout and grow in dilute nutrient mediums such as fish
mucous. Willoughby (1985) developed a ‘rapid preliminary
screening’ system which uses a low nutrient agar to confirm
the existence of S. parasitica by the presence of cysts with
long hairs called "boathooks".

Epidemiology

The disease is reported in early September and late April.

Fish greater than 850-1000 grams in weight under crowded
production ponds are more vulnerable to saprolegniasis. It
causes major losses because of affecting harvestable size
fish (>1 pound) and with very high mortalities among
freshwater catfish facing one or more type of environmental
stresses such as overcrowding, rough handling, transport,
low dissolved oxygen, temperature fluctuations, osmotic
shock and water pollution.
In this disease primary risk factor is inability of fish to
adapt according to rapidly fluctuating water temperatures
during the winter months which impair/imbalances the
fish’s immune system. It causes a loss of mucus from the
skin and temporarily suppresses mucus production by
goblet cells in the dermal layers of the skin. As we know
mucus provides a physical barrier which prevents fungal
spores from contacting and infecting the skin of the fish
and also contains antimicrobial components (including
immunoglobulin or antibodies, lysozyme, complement, C-
reactive protein and proteolytic enzymes) that can destroy
invading zoospores. Skin becomes unprotected without
mucus and fungal spores begin developing masses of fungal
hyphae extending into the muscle tissue. If fungal spores
are not present in sufficient numbers to establish infection
then fish can adapt according to a change in temperature
and regain normal function of goblet cells and mucus
production within 6 days and also can regain their immune
cell function within one month. However, once the infection
is established, fish do not appear to regain normal immune
function and makes the infection more severe. Any
condition that causes a loss of mucus or compromises the
skin or immune system will likely predispose fish to fungal
infections. Physical injuries caused by seining, handling or
crowding, or lesions caused by infectious pathogens create
sites where fungal infection can occur.

The infection may be associated with a significant decrease

in hematological parameters (RBCs, Hb, PCV-Packed Cell
Volume) as well as significant increase in biochemical
parameters (AST, ALT, urea, creatinine, sodium, potassium,
cortisol, insulin and glucose), but iron content decreases
significantly (Zaki et al., 2008).
Symptoms

1. Saprolegniosis in catfishes usually starts as a cotton

wool like white to dark gray or brownish growth over
the head region or dorsal fin and then spread all over
the body and at the tail.
2. The pathological examination revealed a massive
fungal growth resembling a tuft of cotton wool
threads was seen in eyes, gills, fins and in localized
areas of the skin.
3. Microscopically, the fungal hyphae and spores
appeared on eyes, gills, skin and underlying muscles
(muscle rot) with marked degenerative, necrotic and
inflammatory reactions.
4. Brownish patches of cottony fungal growth on the
skin, including the gills
5. Dry depigmented skin and endophthalmia (sunken
eyes).

At first lesions are small, circular, depigmented areas with

hemorrhagic margins. In advanced cases lesions can
become ulcerative; penetrating through the skin and into
the muscle tissue and the fish can be almost completely
covered with thick fungal growth.

Death is thought to be related to the inability of fish to

regulate the salt balance in the blood. It is caused by
sudden decreases in temperature and a significant number
of pathogenic Saprolegnia sp. zoospores in the water (≥ 5
spores/ml).

Treatment and prevention

1. Dip treatment in malachite green (1:10000) for 3

seconds or 5-10 minutes in 3 % NaCl solution or
CuSO4 at the concentration of 5:10000.
2. Pick out the diseased fishes to wash the affected
portion with disinfectant solution of iodine or
K2Cr2O7 (1:10 or 1% solution) afterward a dip
treatment of K2Cr2O7 (1:25000) for 7 days.
3. Maintaining sufficient oxygen concentrations (4 to
5 ppm) may also be important in avoiding
saprolegniasis.
4. Reducing the standing crop of harvestable fish also
will decrease the potential loss from saprolegniasis.
Over densities should not exceed 10000 to 15000
fishes per hectare.
5. Promptly harvesting and selling fish at the very
first sign of saprolegniasis can help to avoid large
losses.
6. Optimizing water quality and reducing stress
especially in the late summer and fall can decrease
the effect of this disease.
7. Pond depth should be high because deeper pond
has more capacity to resist changes in
temperature. Therefore, maintaining ponds at their
maximum depth can reduce temperature
fluctuation and help fish acclimate to changes.
8. 37% Formalin solution is effective in treating
Saprolegnia
9. Hydrogen peroxide is a promising chemical for the
treatment of Saprolegnia with minimal impact to
 the environmental. However, it is important to
consider the species, life stage and water
temperature when treating Saprolegnia with
hydrogen peroxide.
10.Sodium chloride at high concentrations, i.e., sea
water at 29 gm/liter and salt water at 15 gm/liter,
is lethal to Saprolegnia and effective for controlling
S. parasitica, so, it is recommended that a dip bath
of 5 minutes in the solution of NaCl at the rate of
5% .
11.Bath in malachite green @ 5mg/L for one hour.

Tail and fin rot

It is the erosion and disintegrations of fins tails. It is

reported all over the country in cultured fishes. It mainly
occurs before monsoon period. It is more common in ponds
receiving farmyard manure or domestic sewage and cattle
washing

Causative organisms:

It is caused by bacteria Aeromonas hydrophila and

Pseudomonas fluorescens.

Symptoms:

1. At the beginning or early stages more or less distinct

white lines are seen on the marginal surface of the
fins. These lines are actually gill cartilages which
becomes naked after the skin or tissues digestion by
 the pathogens. These lines move toward the base of
the gills and the gills become torn and totally
destroyed and become comb like.
2. It is mainly characterized by erosion and
disintegration of fins and tails of the fishes. Initially
formation of white area on the margins of fins is
noticed which further extends at the base of the fins.
Outer margins become frayed and disintegration of
soft tissue between fin rays starts and continues till
total fin is lost.
3. Epithelial hyperplasia is reported.
4. Excess mucus production.
Transmission: It is transmitted from one pond to other by
transfer of diseased fishes as well as by contaminated
water.

Treatments:

It is treated by using different types of antibiotics and

chemicals at different doses

5. It is treated by Terramycin at the rate of 10-

15 mg / kg body weight of the fish which
mixed with feed.
6. Bath in chloroamphinicol solution of
concentration 60mg/litre at temperature 23-
27°C for 6 days (Mathew, 1961).
7. Dip bath treatment in chemical mixture of
59ml formalin, 59ml of malachite green and
113 liters of water.
8. Dip bath for 1-2 minutes in CuSO 4 solution
of 500 ppm (5mg/litre).
9. In case of serious infection mainly in brooder
fishes the affected parts of fins are removed
surgically and wound is disinfected by
washing with 1 % solution of Silver nitrate
 followed by keeping fish into a solution of
K2Cr2O7 (1:25000) for recovering of wound
for 7days.
Bacterial hemorrhagic septicemia

It is reported to be found all over the country mainly from

coastal areas. It occurs at high temperature and low
dissolved oxygen. Outbreaks are also reported when over
stocking is practiced i.e. overcrowding; fishes during
overcrowding churn or get friction from each other which
causes skin abrasions (the process of wearing away by
friction). It is reported in magur (C. batracus) of 150-200
gms body weight in Haryana.

Causative organisms:

It is caused by bacteria Aeromonas hydrophila and

Pseudomonas fluorescens.

Symptoms:

1. Hemorrhages in skin, fins, oral cavity and muscles

(fig.).
2. The hemorrhage after 3-4 days changes into
ulcer/boil hence its name. Ulcer present on the
epidermis i.e. superficial ulceration.
3. Oozing of blood from the ulcer is also reported during
successive stages of disease.
4. Exophthalmia i.e. outward bulging of eyes. spleen,
liver, kidney and heart multi focal areas of necrosis
 with numerous rod shaped bacteria are reported.

Transmission: It is transmitted via contaminated water or

diseased fish.

Treatments:

1. It is treated with oxytetracycline at the rate of 50-

75mg /kg of body weight for 10-20 days.
2. Cefax 2 litre for one hectare with 40kg CaCO3 in two
equal doses with two days difference can cure this
disease.
3. Desi formula to treat this disease is application of 1
quintal CaO, 10 kg haldi and 1kg KMnO4 per
hectare-meter.
Prevention:

1. Avoid netting in winter

2. Maintain water depth at east 3 to 3.5 feet.

Tejpal Dahiya, Ravi Kant Verma and R.C. Sihag

 Bacterial Diseases Of Fresh Water Fishes

For the maximization of fish yield in a water body, fish

health management is one of the important fish
management strategies. Like other organisms, fishes do
have their pathogens which cause severe health problems,
disorders, impairments and diseases in them. The fish
farmers are the ultimate and most affected lot. Before
undertaking any disease management strategy, there
should be a realistic knowledge about the disease and its
causative organisms.
There are essentially two types of pathogenic bacteria which
are causing fish diseases.

* Primary or obligate pathogens: These are

pathogens that are not part of the normal aquatic flora and
are capable of causing disease in healthy individuals e.g.
Aeromonas salmonicida.

* Opportunistic pathogens: These are normally

free-living, either in the water or in the fish, but are capable
of adopting a pathogenic role under certain circumstances.
Many of these are saprophytes, normally living on dead
organic matter such as plant and animal remains or faeces
e.g. Aeromonas hydrophila, Pseudomonas and Vibrio. The
infections occur in three major ways: a) Damage to the
integument (skin) resulting from an abrasion, parasite
activity or chemical damage (for example from ammonia,
nitrite or high / low pH); b)The fish’s normal defenses are
suppressed as a result of environmental stress or disease.
This enables opportunistic bacteria to take advantage. Fish
can become stressed for a variety of reasons, the most
common being poor water quality, parasites, overcrowding
and thermal stress; c) A substantial increase in the
numbers of opportunistic bacteria due to other infected fish
and /or high levels of decomposing organic matter. This is
typically the situation when the disease starts to spread
and several fish are affected.

Outbreaks of bacterial fish diseases like dropsy,

hemorrhagic septicemia, edwardsiellosis/edwardsiella
septicemia, bacterial kidney disease, bacterial gill disease,
pop eye, vibriosis, epizootic ulcerative syndrome (EUS),
ulcerative and fin and tail rot were reported by different
workers from different parts of the country in freshwater
fishes like catla (Catla catla) , rohu (Labeo rohita), mrigal
(Cirrhinus mrigala), common carp (Cyprinus carpio) and
silver carp (Hypopthalmichthys molitrix) (Kumar et al., 1986;
Karunasagar et al., 1989; Shripal, 2001). These diseases
are reported in farmed as well as in wild fishes. Bacterial
fish diseases play a vital role in determining aquaculture
production. As these are infectious diseases, so prevention
and control of these diseases in aquaculture need early
diagnosis of pathogens. Identification of bacterial fish
pathogens is the primary step in the health management
programme. Lot of research work has been carried out on
pathological studies on the culturable carps.

Bacteria are common infectious agents of fish

diseases and can cause heavy mortality in both cultured
and wild fish (Shotts and Bullock, 1975; Hambal, 1985;
Manasvata, 1985). These are either obligate or facultative
bacterial pathogens. Facultative bacterial pathogens can be
a potential threat when fish are under environmental and
physiological stress (Wedemeyer, 1970). Six gram negative
rods (Aeromonas, Proteus, Citrobacter, Pseudomonas,
Flavobacterium and Chromobacterium) and three gram
positive cocci (Micrococcus, Streptococcus and
Staphyllococcus) genera of bacteria which are potentially
pathogenic were identified from Aristichthys nobilis and
Ctenopharyngos idella fingerlings (Shamsudin, 1986).

Bacterial swabs were taken aseptically from the lesions,

kidney and liver of the diseased fishes and cultured on the
selective media. Aeromonas Agar Base (Oroid)
supplemented with antibiotic C-F-C (Centrimide- Fucidin-
Cephaloridine) SR 103E and SS Agar. These first two
selective media were found to be suitable for isolation and
determination of aeromonads and pseudomonads,
respectively (Chowdhury and Inglis, 1994a, 1994b). Pure
cultures of Aeromonas spp., Pseudomonas spp. and
Edwardsiella spp. were obtained from selective agar culture
plate. These isolates were maintained in Tryptone Soya
Agar ( TSA) slant at 4ºC. Selective bacterial pathogens were
primarily identified up to genus level. In order to identify
bacterial isolates, API – 20E test kit procedure was followed
in addition to the conventional methods described in Cowan
and Steel’s Manual for the identification of medical bacteria
after Barrow and Feltham (1993). However, species of
Aeromonas were confirmed after Carnahan et al. (1991).

The aeromonads were short, gram negative, motile

bacilli with a single flagellum that fermented glucose with
or without production of gas. Snieszko (1957) later divided
the genus into three species: A. hydrophila, A. punctata and
A. liquifaciens. Schubert (1967) confirmed that there
was enough biochemical similarity to establish the genus
Aeromonas, but invalidated species – species distinction.
Later, Popff and Vernon (1976) demonstrated that the
motile aeromonads could be classified into two distinct
species: A. hydrophila (comprised of organisms previously
described as A. punctata and A. liquifaciens), and a new
species A. sorbia. Biochemically, A. hydrophila hydrolyzes
esculin and ferments both salicin and arabinose, whereas
A. sorbia does not utilize these compounds (Lallier et al.,
1981). Motile aeromonads may be pleomorphic but
generally produce circular and smooth raised colonies on
agar. Hsu et al. (1985) noted that all the isolates of motile
aeromonads they studied produced acid from fructose,
galactose, maltose, mannitol, trehalose, dextrin and
glycogen; 99.4% of the strains produced acid from glucose,
98.8% from mannose and 98.2% from glycerol. Acid
production from other carbohydrates (arabinose, salicin,
cellobiose, sucrose and lactose) varied.
 Shotts et al. (1985) also found that all A. hydrophila
complex strains hydrolyzed albumin, casein and fibrinogen;
most strains also digested gelatin (99.9%), hemoglobin
(94.3%) and elastin (73.2%) but none of the strains
hydrolyzed collagen. Further phenotypic differentiation of
the six (A. hydrophila, A. veronii, A. caciae, A. scubertii, A.
janddaei and A. trota.) most commonly isolated motile
aeromonads obtained from clinically affected fishes (Joseph
and Carnahan, 1994). They described that all bacterial
isolates would be short gram negative, oxidase – positive
bacilli that ferment glucose and are resistant to the
vibriostatic agent (0129). However, Austin et al. (1989) have
shown that A. hydrophila, A. sobria and A. caviae comprise
the most predominant clinical isolates that are typically
associated with fish.

EUS is a dreaded fish disease of the Asia-Pecific

region. It broke out in India, for the first time, in 1988 and
since then India has witnessed the gradual spread of the
disease in fresh water and also in brackish water fishes in
all types of water bodies in different states of the country.
About 30 species of fresh water and brackish water fishes
in India have been found infected with this disease (Das,
1997). The freshwater fishes affected with EUS in India are
Catla catla, Labeo rohita, Cirrhinus mrigala, Puntius
javanicus, Ctenopharyndon idella and Hypophthalmichthys
motitrix. The brackishwater fishes affected with EUS are
Liza persia, L. cephalus, L. subviridis and Etroplus sp.
likewise, some wild varieties of fish species of fresh water
are Channa striatis, C. punctatus, C. gachaua, Clarius
batrachus, Heteropneustres fossilis and Mystus vittatus.

A number of bacteria viz. A. punctatus,

Flavobacterium sp., Pseudomonas sp. Edwardsiella tarda,
Vibrio parahaemolyticus and Streptococcus sp. have been
isolated from the affected specimens (Kumar, 1989).Besides
bacteria, viruses, fungi and parasites were also reported to
be associated with epizootic ulcerative syndrome (Kumar et
al., 1991; Dey, 1989, Chowdhury et al., 2003). Studies
revealed complete loss of epidermis in the ulcerative area of
the skin where hypodermis showed characteristic
granulomatous changes (Dey, 1989). Gill showed
hyperplasia of the secondary lamellar epithelium, fusion of
lamella and clubbing of gill tips. In some cases oedema and
hypotrophin changes were also observed (Dey, 1989).

Ahmed and Hogue (1999) performed a

histopathological study on five species of fresh water fishes
(Cirrhinus mrigala, Catla catla, Puntius sophore,
Glossogobius guiris and Labeo rohita) affected by EUS in
Bangladesh. Mycotic (fungal) involvement was observed by
them in EUS affected fish species. Clinically and
histologically C. mrigala was observed to be the most
severely affected species by EUS, followed by C. catla, P.
sophore and Glossogobius guiris. L. rohita was found to be
the least affected species.

Histopathology of EUS affected fishes has been

studied by various workers in different species of fishes in
India and abroad (Roberts, 1997; Dey, 1989; Das, 1992;
Kumar et al., 1991; Mukherjee, 1991; Nayak, 1993; Das et
al., 1990). The changes were characterized by cellular
infiltration in the hypodermis and epidermis, oedema,
dilation and necrosis of primary and secondary lamellae of
the gill tubular epithelial cells, degeneration of kidney
tubules, depletion of lymphoid cells of spleen and
congestion of cerebral tissue. Most of the hepatocytes
showed cloudy swellings. In case of Chinese carp (grass
carp and common carp), the affected fish showed ulcer and
deep red hemorrhagic lesions were found in the region of
caudal peduncles.

Ecological studies on Aeromonas sp. and

Pseudomonas sp. in mrigal in cultured ponds were
conducted by Iqbal et al. (1999). EUS suffered fish samples
were found to have high percentage of bacterial species.

Rahman et al. (2004) investigated distribution

pattern of a collection of 106 Aeromonas strains isolated
from both healthy and EUS affected fish, septicemia disease
affected frog and turtle in Bangladesh, Japan, Malaysia,
Philippines. The study was conducted on physico-
biochemical characterization of strains and subsequent
confirmation by analysis of the 16S rRNA sequences of
some randomly chosen representative strains from all
identified phenotype. Special emphasis was given to confirm
a group of strains, which belonged to a newly identified
species, Aeromonas sp. T8 group was particularly found in
different species of EUS-affected fish in Philippines and
Thailand. A. hydrophila subsp. hydrophila and A.
hydrophila subsp. ranae were recovered from EUS affected
fish and septicemic disease of frog and turtle. A. hydrophila
subsp. hydrophila was distributed in Bangladesh and
Thailand while A. hydrophila subsp. ranae was found only
in Thailand. A. veronii biotype sobria and A. veronii biotype
veronii was found to be dispensed mostly in EUS affected
fish in different countries. A. jandaei was obtained from
EUS-positive fish in Bangladesh and Malaysia but A. media
from healthy fish in Bangladesh.

Filamentous, non gliding bacterium strains of

Flavobacterium sp. were isolated from sheat fish (Silurus
glanis), silver carp (Hypohpthalmichys molitrix) and rain
bow trout (Salmo gairdneri) affected by cold water gill
disease in Hungarian fish culture (Farkas, 1985). There are
only two reports on filamentous, non gliding or swarming
fish pathogens. The first observation was made in Japan by
Kimura et al. (1978) in connection with gill disease of
salmonid fish. Wakabayashi et al. (1980) compared the
strains originating from Japan with strains isolated from
Oregon, USA. The salmonid fish seemed weak with slow
movements and signs of oxygen deficiency. The gill did not
loose its colour nor was any necrosis observed.

Dropsy is an abnormal accumulation of fluid in the

body of an animal which may either affect the whole body
or be localized in some organs or tissues (Gopalakrishan,
1961). After the first report of dropsy by Schaperclaus
(1930), lot of investigations has been carried out in India
(Gopalkrishan 1961; Kumar et al., 1986, Sahoo et al.,
1998). Gopalkrishan (1961) found only Aeromonas sp. as
causative agent whereas Kumar et al. (1986 a,b) found
mixed infection of Aeromonas hydrophila and Myxosporidian
sp. in dropsy of carps. Dropsy of carp also (called carp
rubella) is also considered to show viral etiology
(Schaperclaus, 1965; Fijan et al., 1971). The various forms
(latent, ascitic, sub acute and chronic) of the disease infact
complicate the diagnosis of disease (Gaines and Rogers,
1975).

Kluyner and Van Niel (1936) renamed many

organisms that were associated with hemorrhagic
septicemias in fish within the genera Bacillus,
Pseudomonas, Proteus and Aerobactor into the new genus
Aeromonas. These aeromonads were short, gram negative,
motile bacilli with a single flagellum that fermented glucose
with or without the production of gas.

Motile aeromonads can also cause disease in warm

blooded vertebrates. A. hydrophila may cause endocarditis,
septioarthritis, diarrhea, corneal ulcers, skin and wound
infections, meningitis and fulminating septicemias (Von
Gravenitz and Mensch 1968; Davis et al.,1978). Clinical
isolates have been obtained from retail foods (fish, seafood,
raw milk, poultry and red meat) and all isolates had
biotypes identical to those of enterotoxin – positive strains
(Palumbo et al., 1989). The ability of these bacteria to grow
competitively at 5ºC may be indicative of their potential as a
public health hazard. However, most A. hydrophila were
reduced to non- detectable levels on catfish filets cooked to
70ºC (Huang et al., 1993).

Jhingran (1991) observed fin and tail rot disease

which affected both adults and young cultivable fishes. The
infection during its early stage appear as white line on the
margin of fin, spreading an imparting frayed appearance on
the appendages which later on eventually putrefies and
disintegrates. This disease is contagious and may cause a
considerable damage. Fin and tail rot disease was reported
in large number of fresh water fishes in India (Kumar et al.,
1986; Karunasagar et al., 1989).

Walters and Plumb (1980) and Kumar et al. (1986)

have recorded enlargement of kidney tubules and
hemorrhages and necrosis due to A. hydrophila infection.
Ventura and Grizzle (1988), Kumar et al.(1991), Das (1991)
and Nayak (1993) while studying histopathological changes
had described degeneration and vaculation of the tuber
cells and necrosis of kidney affected with A. hydrophila.

Das and Das (1997) have discussed that pH above

9.0 may have sublethal affects on many fish species of
culturable varieties. The pH value at 11.0 lethal to all
species of fish and prawn.

Das and Das (1993) observed that the total hardness

concentration in majority of culturable water area should
be in optimum level of ranges between 140 ppm to 400 ppm
and above this range creates health problems and
infectious in culturable species of fishes. They also
observed that high external concentration of CO 2 interferes
oxygen uptake and causes respiratory problem and at last
leads to gill necrosis.

Boyd (1974) revealed that fluctuations in the

alkalinity level may be directly harmful to fish populations
and leads to infection in ponds if the level of alkalinity is
high but if the level of alkalinity is greater than 300 mg/l,
the pond will become unproductive because of limitation of
CO2 availability at such a high concentration.

The fore going paragraphs reveal that large number

of fish disease

causing organisms are found in different water bodies in

India and other parts of the globe. Only symptomatic
studies have been made in India to identify these disease
and their probable causative organisms. However, now
there are several biochemical tests which can confirm the
presence of specific causative organisms (Kreig and Holt,
1984).

A software programme for identification of bacterial

isolates has been developed (Bryant, 1995, 2004; Carson et
al., 2001; Ottaviani et al., 2003 and Willcox et al., 1973). By
this programme probabilistic identification matrix of
bacteria (PIBWin) was calculated and near probability of
occurrence of causative organism is determined by
calculating its ID score. This latest methodology has been
reviewed and used in the present investigations.

Tejpal Dahiya, Ravi Kant Verma and V.P. Saini

 Probiotics In Aquaculture
Aquaculture is an important sub-sector of agriculture. It
has great potential and plays a significant role in the
national economy by contributing towards food security of
the country by reducing pressure on demand for mutton,
beef and poultry and by earning foreign exchange.
Aquaculture has been established as an industry in India in
the last decade. About 7.5% of total annual income of the
country comes from aquaculture.

Disease outbreaks are being increasingly recognized

as a significant constraint on aquaculture production and
trade affecting the economic development of the sector in
many countries. So far, conventional approaches such as
the use of disinfectants and antimicrobial drugs, have had
limited success in the prevention or cure of aquatic disease
(Subasinghe, 1997). Combined with the problem of
antibiotic contamination of aquaculture facilities and
livestock, the indiscriminate worldwide use of antibiotics in
aquaculture has led to the development of drug-resistant
bacteria which are becoming increasingly difficult to control
and eradicate. Furthermore, there is a growing concern
about the use and, particularly, the abuse of antimicrobial
drugs not only in human medicine and agriculture but also
in aquaculture. The massive use of antimicrobials for
disease control and growth promotion in animals increases
the selective pressure exerted on the microbial world and
encourages the natural emergence of bacterial resistance
(World Health Organization antimicrobial resistance fact
sheet 194, http://www.who.int/inf-fs/en/fact194.html).
Not only can resistant bacteria proliferate after an antibiotic
has killed off the other bacteria, but also they can transfer
their resistance genes to other bacteria that have never
been exposed to the antibiotic.

As a result of resistant bacterial strains becoming

more prevalent and difficult to treat, alternative methods of
controlling the microbial environment are being
investigated. One of the methods gaining recognition for
controlling pathogens within the aquaculture industry is
the use of beneficial or probiotic bacteria (Ringø and
Gatesoupe, 1998; Verschuere et al., 2000; Irianto and
Austin, 2002).

The term, probiotic, simply means “for life”,

originating from the Greek words “pro” and “bios”
(Gismondo et al., 1999). The original definition (Lilly and
Stillwell, 1965) of probiotics “substances produced by one
protozoan that stimulated the growth of another” was
expanded on from an agricultural perspective and defined
as “a live microbial feed supplement which beneficially
affects the host animal by improving its intestinal microbial
balance” (Fuller, 1989). Gatesoupe (1999) redefined
probiotics for aquaculture as “microbial cells that are
administered in such a way as to enter the gastrointestinal
tract and to be kept alive, with the aim of improving
health”. The definition provided by Verschuere et al. (2000)
includes, 1) the ability of a probiotic to modify the “host-
associated or ambient microbial community” and, 2) to
improve the quality of its surroundings, both of which can
be considered as biocontrol. Apart from the requirement of
the probiotic to be a live culture, this definition is a lengthy
way of describing a probiotic as defined by Irianto and
Austin (2002) thus “a probiotic is an entire or
components(s) of a microorganism that is beneficial to the
health of the host”.

The concept of probiotic activity has its origins in the

knowledge that active modulation of the gastrointestinal
tract (GIT) could confer antagonism against pathogens, help
development of the immune system, provide nutritional
benefits and assist the intestinal mucosal barrier (Vaughan
et al., 2002). Today probiotics are quite commonplace in
health promoting “functional foods” for humans, as well as
therapeutic, prophylactic and growth supplements in
animal production and human health (Senok et al., 2005).
The research of probiotics for aquatic animals is increasing
with the demand for environment friendly aquaculture. The
gastrointestinal microbiota of fish and shellfish are
peculiarly dependent on the external environment, due to
the water flow passing through the digestive tract. Most
bacterial cells are transient in the gut, with continuous
intrusion of microbes coming from water and food. Most
attempts to propose probiotics have been undertaken by
isolating and selecting strains from aquatic environment.
These microbes are Vibrionaceae, pseudomonads, lactic
acid bacteria, Bacillus spp. and yeasts.

Typically, the lactic acid bacteria (LAB) have been

widely used and researched for human and terrestrial
animal purposes, and LAB are also known to be present in
the intestine of healthy fish (Hagi et al., 2004). Interest in
LAB stems from the fact that they are natural residents of
the human GIT with the ability to tolerate the acidic and
bile environment of the intestinal tract. LAB also function to
convert lactose into lactic acid, thereby reducing the pH in
the GIT and naturally preventing the colonization by many
bacteria (Mombelli and Gismondo, 2000). The most widely
researched and used LAB are the lactobacilli and
bifidobacteria (Senok et al., 2005). Other commonly studied
probiotics include the spore forming Bacillus spp. and
yeasts. Bacillus spp. have been shown to possess adhesion
abilities, produce bacteriocins (antimicrobial peptides) and
provide immunostimulation (Duc et al., 2004; Barbosa et
al., 2005). The strains appear to be effective probiotics and
commercial products containing such strains have been
demonstrated to improve shrimp production to a level
similar to that when antimicrobials are used. Bacillus spp.
hold added interest in probiotics as they can be kept in the
spore form and therefore stored indefinitely on the shelf
(Hong et al., 2005).

Non-bacterial sources of probiotics

The yeast, Saccharomyces cerevisiae, also has been

commonly studied whereby immunostimulatory activity was
demonstrated and production of inhibitory substances shown
( Dahan et al., 2003).

Yeasts have been identified which exhibit probiotic

characteristics. Yeasts are not affected by antibiotics. This
is advantageous in probiotic preparations used for
preventing disturbances in the normal microflora in the
presence of antibacterial metabolites. Strains of S.
cerevisiae and D. hansenii isolated from salmonids have
been shown to attach and grow in fish intestinal mucus.
A list of probiotic bacteria used commercially is given
below.

Lactobacillus species

L. acidophilus

L. casei

L. fermentum

L. gasseri

L. johnsonii

L. lactis

L. paracasei

L. plantarum

L. reuteri

L. rhamnosus

L. salivarius

Bifidobacterium species

B. bifidum

B. breve

B. lactis

B. longum

Streptococcus species

S. thermophilus

Probiotic criteria
 In general, the selection criteria for a bacterium to be used
as a ‘probiotic' should be:

 non-pathogenic;
 withstand incorporation into a delivery vehicle at high
cell counts, and
remain viable throughout the shelf-life of the product;
 withstand transit through the gastrointestinal tract, that
is, show acid and bile tolerance;
 be able to adhere to cells of the intestinal epithelium
and/or colonize the lumen of the tract;
 show antagonistic activity towards enteric pathogens
and/or
provide demonstrated health benefits

Properties of probiotics

It has been widely published that a probiotic must

possess certain properties (Verschuere et al., 2000). These
properties were proposed in order to aid in correct
establishment of new, effective and safe products. The
properties include:

1. The probiotic should not be harmful to the host it is

desired for,
2. It should be accepted by the host, e.g. through
ingestion and potential colonization and replication within
the host,
3. It should reach the location where the effect is
required to take place,
4. It should actually work in vivo as opposed to in vitro
findings,
5. It should preferably not contain virulence resistance
genes or antibiotic resistance genes.
Possible modes of action
Production of Antagonistic Compounds

Antagonistic compounds are described as chemical

substances produced by bacteria which are toxic or
inhibitory towards other micro organisms. These
substances may be produced as either primary or
secondary metabolites and therefore have different modes of
inhibitory action. Microbial populations may release
chemical substances that have a bactericidal or
bacteriostatic effect on other microbial populations, which
can alter interpopulation relationships by influencing the
outcome of competition for chemicals or available energy
(Fredrickson and Stephanopoulos, 1981). The presence of
bacteria producing inhibitory substances in the intestine of
the host, on its surface, or in its culture medium is thought
to constitute a barrier against the proliferation of
(opportunistic) pathogens.

In general, the antibacterial effect of bacteria is due

to the following factors, either singly or in combination:
production of antibiotics (Williams and Vickers, 1986),
bacteriocins (Bruno and Montville, 1993), siderophores,
lysozymes, proteases, and/or hydrogen peroxide and the
alteration of pH values by the production of organic acids
(Sugita et al, 1997). Lactic acid bacteria are known to
produce compounds such as bacteriocins that inhibit the
growth of other microorganisms (Vandenbergh, 1993). There
are many reports of the inhibitory activity of lactic acid
bacteria mediated by bacteriocins, mostly but not
exclusively against gram-positive bacteria. Nair et al. 1985
showed that a large proportion of marine bacteria produced
bacteriolytic enzymes against V. parahaemolyticus. Other
inhibitory compounds produced by bacteria include organic
acids, hydrogen peroxide, carbon dioxide and siderophores .
Bacteriocins are proteinacious agents produced by bacteria
to inhibit or kill other bacteria. The genes encoding the
proteins involved in processing the bacteriocin are often
carried out by plasmids or transposons suggesting that the
ability to produce bacteriocins can be transferred between
bacteria. Siderophores are iron-complexing chemicals
secreted by bacteria and fungi. Bacteria which produce
siderophores have highly specific transport proteins in their
3+
outer membrane which allow them to utilise Fe which is
otherwise insoluble in the surrounding medium.
Siderophore-producing bacteria can survive in nutrient-
poor environments

Competition for Chemicals or Available Energy

Competition for chemicals or available energy may

determine how different microbial populations coexist in the
same ecosystem (Fredrickson and Stephanopoulos, 1981).
The microbial ecosystem in aquaculture environments is
generally dominated by heterotrophs competing for organic
substrates as both carbon and energy sources. Specific
knowledge of the factors governing the composition of the
microbiota in aquaculture systems is required to
manipulate it. Verschuere et al., 1999 selected several
strains with a positive effect on the survival and growth of
Artemia juveniles. The in vitro antagonism tests and filtrate
experiments showed that no extracellular inhibitory
compounds were involved in the protective action of these
strains against V. proteolytics CW8T2 but that living cells
were required to protect Artemia against the pathogen. It
was suggested that the selected bacteria exerted their
protective action by competing with the pathogen for
chemicals and available energy.

Competition for iron

Virtually all microorganisms require iron for growth

(Reid et al, 1993). Siderophores are low-molecular-weight
(<1,500), ferric ion-specific chelating agents which can
dissolve precipitated iron and make it available for microbial
growth. The ecological significance of siderophores resides
in their capacity to scavenge an essential nutrient from the
environment and deprive competitors of it. Successful
bacterial pathogens are able to compete successfully for
iron in the highly iron-stressed environment of the tissues
and body fluids of the host. The ecological significance of
siderophores in soils as important tools for iron acquisition
by microorganisms and plants and their involvement in
suppression of plant root pathogens have been established.
Harmless bacteria which can produce siderophores could
be used as probiotics to compete with pathogens whose
pathogenicity is known to be due to siderophore production
and competition for iron or to outcompete all kind of
organisms requiring ferric iron from solution. The possible
effectiveness of siderophore-producing probiotics can be
illustrated by the study of Gatesoupe (1997), in which the
addition of the bacterial siderophore deferoxamine to live
food (rotifers) increased the resistance of turbot larvae
challenged with the pathogenic Vibrio strain P. The addition
of a siderophore-producing Vibrio strain E protected the
turbot larvae slightly more.

Competition for Adhesion Sites

One possible mechanism for preventing colonization

by pathogens is competition for adhesion sites on gut or
other tissue surfaces. It is known that the ability to adhere
to enteric mucus and wall surfaces is necessary for bacteria
to become established in fish intestines (Vesterdahl et al,
1991). Since bacterial adhesion to tissue surface is
important during the initial stages of pathogenic infection (,
competition for adhesion receptors with pathogens might be
the first probiotic effect. Adhesion can be nonspecific, based
on physicochemical factors, or specific, involving adhesin
molecules on the surface of adherent bacteria and receptor
molecules on epithelial cells. Inhibition of the adhesion of
pathogens to human or other mammal cells has already
been demonstrated in vitro by several authors . Adhesion
capacity and growth on or in intestinal or external mucus
has been demonstrated in vitro for fish pathogens like
V. anguillarum and A. hydrophila (Krovacek et al, 1987)
and for candidate probiotics such as Carnobacterium strain
K1 and several isolates inhibitory to V. anguillarum . In
one of these studies, the aim was to measure the in vitro
capacity of the strains to adhere to and grow in turbot
intestinal mucus in order to investigate their potential to
colonize farmed turbot as a means of protecting the host
from infection by V. anguillarum (Olsson et al, 1982). The
intestinal isolates generally adhered much better to a film of
turbot intestinal mucus, skin mucus, and bovine serum
albumin than did V. anguillarum, indicating that they could
compete effectively with the pathogen for adhesion sites on
the mucosal intestinal surface.

Enhancement of the Immune Response

Immunostimulants are chemical compounds that

activate the immune systems of animals and render them
more resistant to infections by viruses, bacteria, fungi, and
parasites (Raa, 1996). Fish larvae, shrimps, and other
invertebrates have immune systems that are less well
developed than adult fish and are dependent primarily on
nonspecific immune responses for their resistance to
infection. Observations obtained in experiments with warm-
blooded animals indicate that probiotic (lactic acid) bacteria
administered orally may induce increased resistance to
enteric infections. It has also been suggested that ingestion
of bacteria and subsequent endocytosis in cod and herring
larvae are involved in stimulation of the developing immune
system.
Improvement of Water Quality

In several studies, water quality has been recorded

during the addition of the probiotics, especially Bacillus
spp. The rationale is that gram-positive Bacillus spp. are
generally more efficient in converting organic matter back to
CO2 than are gram-negative bacteria, which would convert a
greater percentage of organic carbon to bacterial biomass or
slime (Stanier, 1963). It is reasoned that by maintaining
higher levels of these gram-positive bacteria in the
production pond, farmers can minimize the buildup of
dissolved and particulate organic carbon during the culture
cycle while promoting more stable phytoplankton blooms
through the increased production of CO2.

Interaction with Phytoplankton

Recent reports demonstrate that many bacterial

strains may have a significant algicidal effect on many
species of microalgae, particularly of red tide plankton
(Fukami et al, 1997). Of 41 bacterial strains tested,
23 inhibited the growth of the unicellular alga Pavlova
lutheri to various degrees (Munro et al., 1995). Bacteria
antagonistic toward algae would be undesirable in larval
rearing where unicellular algae are added (e.g., the green-
water technique) but would be advantageous when
undesired algal species develop in the culture pond. Positive
effects of bacteria on cultured microalgae have also been
observed (Fukami et al, 1997). It is conceivable that bacteria
can indirectly influence the health or the zootechnical
performance of the cultured aquatic animals through their
effect on the microalgae used as food or in the green-water
technique. When probiotic bacteria are selected to be used
in a culture environment comprising algae, their possible
interaction with these unicellular algae must be taken into
consideration when the mode of action is being investigated.
Mode of Application of the Probiotics

The probiotics can be added to the host or its ambient

environment in several ways:

1) Addition to the artificial diet, 2) Addition to the

culture water, & 3) Bathing and addition via live food
(Verschuere et al., 2000; Gomez-Gil et al., 2000).

The concept of inoculating in the culture system and

adding to live food is advisable when we use small volumes.
The inclusion in the artificial diet is more adapted when
using greater volumes; however, this requires of sufficient
number of probiotics that last when they reach the
gastrointestinal tract and present capacity to adhere to the
epithelial mucosa.
The development of a probiotic selection protocol
Figure - Proposed research protocol for the selection of
intestinal probiotics in marine larviculture (Verschuere
et al., 2000).

Acquisition of Background Information

Before research and development activities are

begun, bottlenecks in culture practices or in economic
development of the aquaculture farm or industry should be
identified, taking into account possible restricting or
directing legislative decisions. A critical review of the
available scientific literature and a profound knowledge of
the rearing practices used in aquaculture farms should
determine whether a probiotic approach would be feasible
and is worth further examination. The abiotic and biotic
environment inhabited by the cultured organisms should be
well characterized, with particular attention being paid to
the microbiota, the relationships between the microbiota
(pathogens and others) and the host, and the relationships
between the microbiota and the carrying capacity of the
culture environment.

Acquisition of Putative Probiotics

The acquisition of a good pool of candidate probiotics

is of major importance in this process. It is vital in this
phase that the choice of strains is made as a function of the
possible role of the probiotics to be developed, although
there is no unequivocal indication that putative probiotics
isolated from the host or from their ambient environment
perform better than isolates completely alien to the cultured
species or originating from a very different habitat. It is
assumed that strains showing a dominant colonization of,
e.g., the intestinal mucus of fish are good candidates to
competitively exclude pathogens from the adhesion sites of
the gut wall. Similarly, the presence of a dominant bacterial
strain in high densities in culture water indicates its ability
to grow successfully under the prevailing conditions, and
one can expect that this strain will compete efficiently for
nutrients with possibly deleterious strains. Identification of
the isolates at this stage is not essential.

Screening and Preselection of Putative Probiotics

Generally at the end of the previous phase, one ends

up with a pool of isolates that must be screened and
preselected to obtain a restricted number of isolates for
further examination. Several approaches followed in
literature are discussed below. The application of in vitro
tests to screen the acquired bacterial strains presupposes a
well-known mode of action to select the appropriate in vitro
test. Since the evidence about the possible modes of action
of probiotics is still equivocal, preference should be given to
in vivo tests in the search for probiotics. The use of the
target organism in the screening procedure provides a
stronger basis.

In vitro antagonism tests

A common way to screen the candidate probiotics is

to perform in vitro antagonism tests, in which pathogens
are exposed to the candidate probiotics or their extracellular
products in liquid (Gram et al, 1999) or solid (Austin et al,
1992) medium. Depending on the exact arrangement of the
tests, candidate probiotics can be selected based on the
production of inhibitory compounds (Gram et al, 1999) or
siderophores, or on the competition for nutrients (Dopazo et
al, 1998). The preselection of candidate probionts based on
these in vitro antagonism tests has often led to the finding
of effective probiotics.

Colonization and adhesion

It is stated above that a candidate probiotic should

either be supplied on a regular basis or be able to colonize
and persist in the host or in its ambient environment. The
ability of a strain to colonize the gut or an external surface
of the host and adhere to the mucus layer may be a good
criterion for preselection among the putative probiotics.
This involves the viability of the potential probiotic within
the host and/or within its culture environment, adherence
to host surfaces, and the ability to prevent the
establishment of potentially pathogenic bacteria.

Evaluation of Pathogenicity of Selected Strains

Before a culture can be used as a probiotic, it is

necessary to confirm that no pathogenic effects can occur
in the host. Therefore, the target species should be
challenged with the candidate probiotic, under normal or
stress conditions. This can be done by injection challenges,
by bathing the host in a suspension of the candidate
probiotic, or by adding the probiotic to the culture.
Garriques and Arevalo (1995) examined the pathogenicity of
three bacterial isolates toward Litopenaeus vannamei by
adding these to the nauplii cultures. Mortality was recorded
for up to 4 days, and the animals were monitored for
phototactic response. This phase in the search for
probiotics can be combined with the previous one when
small-scale in vivo tests are performed.

In Vivo Evaluation of Potential Probiotic Effects on the

Host
 The effect of candidate probiotics should be tested in
vivo as well. When the probiotic effect is supposed to be
nutritional, the candidate probiotics could be added to the
culture of the aquatic species and their effect on growth
and/or survival parameters could be assessed. However,
when biological control of the microbiota is desired,
representative in vivo challenge tests seem to be the
appropriate tool to evaluate the potential effect of the
candidate probiotic on the host.

Experimental infections in vivo antagonism tests

The next important step in the in vivo challenge test

involves experimental infection with a representative
pathogen. Pathogens or opportunistic pathogens can be
administered via the diet (live food or artificial), through
immersion, or via the culture water, similarly to the
probiotics. In experiments performed by Gildberg et al.
(1995), Atlantic salmon fry was challenged with cohabitant
fishes which were previously infected with Aeromonas
salmonicida through intraperitoneal injection. The mortality
of the challenged fishes due to acute furunculosis was
monitored over time.

Mass Production, Economic Evaluation, and Evaluation

of Compliance with legislation

The approach outlined above should result in a set of

strains with a well-established probiotic effect on the target
organism without affecting other possibly involved trophic
levels. Comparative pilot experiments under hatchery or
grow out conditions in the farms should be performed to
estimate the economic consequences of the probiotic
application. An important factor in the economic evaluation
is the mass production of the probiont. Also, effective
legislation, if any, should be taken into account before
commercial application is begun. Finally, a cost-benefit
analysis will determine whether the probiotics could be
applied in practice or not.

Conclusion

The application of probiotics in aquaculture shows

promise, but needs considerable efforts of research. The
first question, unanswered in many cases, is the fate of the
probiotic in rearing medium and in gastrointestinal tract.
Immunological and molecular probes will be useful tools to
trace the probiotic cell. It is essential to investigate the best
way of introduction and the optimal dose, and technical
solutions are required, especially to keep the probiotic alive
in dry pellets. The spores of Bacillus spp. are especially easy
to introduce in dry food, and this is an additional advantage
of these promising candidate probiotics (Rengpipat et al.,
1998; Sugita et al., 1998). Lactic acid bacteria are also good
candidates, and further studies are necessary to evaluate
the interest of yeasts as probiotics. The bacteria normally
dominant in healthy animals may be sources of probiotics,
but there are many potential pathogens among
Vibrionaceae and pseudomonads. It may be wise to carry
out long-term surveys to make sure that the bacteria keep
innocuous, without risk of apparition of potentially
detrimental mutants. The influence of probiotics on
gastrointestinal microbiota remains poorly described, but
further investigation may be expected with the propagation
of molecular approaches to analyse bacterial communities
(Raskin et al., 1997; Hugenholtz et al., 1998).

Tejpal Dahiya, Ravi Kant Verma and R.C. Sihag

 Fish Immunology
Fish hatcheries and farms are becoming increasingly
important in supplementing food production. However in
most of the facilities the animals are kept at relatively high
densities causing problems of stress and diseases but it has
been seen that disease and stress problems have grown
proportionally with the intensive culture of aquaculture
species and main reasons are :

1. Increased stocking densities.

2. Broodstock acts as infected carriers.

3. General management practices are followed without

giving importance to hygiene.

4. Nutrition is poor; there is presence of antinutritional

factors in feed.

5. Substandard water quality

All the factors cause great susceptibility to disease

and stress via weakening of resistance under intensive
culture conditions. Fish, under normal condition are more
susceptible to disease. Teleosts are of considerable
economic importance thus an understanding of its immune
system i.e. their ability to resist infection is of great interest
to fisheries.

The state of body in which there is impairment or

interference with its normal functioning due to one or the
other reason is termed as disease and any substance which
causes a disease by its excess or deficiency or absence is
called a disease agent which may be biological agent,
chemical, nutritive, physical or mechanical agent. Biological
agents are also called as pathogens which include bacteria,
viruses, fungi, protozoans etc. To defend against these
pathogens the organism needs immune system which is
responsible for defense mechanism. Before discussing
about defense mechanism and immune system let us
discuss about antigens and antibodies.

Any foreign substance which when enters inside the

body inside the body and is capable of stimulating an
immune response is called an antigen while the protective
chemicals produced by the body in response to antigens are
called antibodies. So the antigens are also called as
antibody generating chemicals or immunogens and
antibodies are called immunoglobulins (Igs).

In general, vetebrates have five types of antibodies on

the basis of their location, physical properties and function
i.e. IgG, IgA, IgM, IgD, IgE. Electron microscopic studies
have revealed that IgG is a Y shaped tetrapeptide protein
formed of 2 light chains and two heavy chains which are
identical.

Light chains :

It is formed of about 214 amino acids. It is of 2 types

in all class of immunoglobulins i.e. Kappa (K) and Lambda
(l) chains which differ from each other in their primary
amino acid sequence. A given antibody has either both
kappa and both lambda type light chains. Light chain is
formed of two parts : Variable region and constant region.
Variable region has 1-108 amino acids from N terminal end
and varies from one antibody to other and the constant
region has 109-214 amino acid. Each chain has two intra
chain disulphide (S-S) loops, one in variable region between
cystine residues at 23-88 position and other in constant
part between cystine residues at 134 and 194 positions.

Heavy chains:
These are 2 in number. Each is formed of 440 amino
acids. It is formed of two parts variable region formed of 1-
118 amino acids from N terminal and constant region is
formed of 119-440 aa and is 3 times in length to that of
chain. Constant region each heavy chain has three
interaction S-S loops which divide the part into 3 regions.

CH1 - 144 - 200

CH2 - 261 - 322 Receptor to bind complement

CH3 - 367 - 425 Receptor to bind macrophage

22.96 position S-S loop in variable region.

The part of hyper variable region on antibody which

binds the antigen is called paratope while corresponding
part of antigen which is in contact with paratope is called
epitope.

Hypervariable spots situated between different amino

acids are :

Light chain Heavy

chains

24-34 31-35
 50-56 50-65

89-97 81-85

95-102
Antibody molecule has image region rich in proline
residue. There is flexibility around hinge region. It has 2
antigen binding fragments (F ab) involving variable region of
both light and heavy chains.
There are 4 subclasses of human IgG i.e. IgG 1, IgG2,
IgG3, IgG4 and 2 subclasses of human IgA i.e. IgA1, and
IGA2.

In fishes only IgM has been identified.

Ig(M) molecule is assembled without the uniform

disulfide cross-linking of all monomeric subunits. Random
disulfide cross-linking generates a pool of IgM molecules
with a variety of structurally different tetramers termed
redox form. These tetramers possess varying degree of
disulphide polymerization, where some are completely
polymerized while others possess only 2 or 3 disulphide
bond. Each antibody producing all assembles all of these
different forms. Thus all different redox forms can share
identified sites.

Antigen-antibody interactions :

Each antibody is specific for a particular antigen due

to unique structural organization of amino acids in variable
portions of both light and heavy chains. Antibodies have a
different stearic shape for each antigen specificity so when
an antigen comes in contact with an antibody, the
prosthetic groups of antigen fit as a mirror image with those
of antibody.
 Antibodies act in 2 different ways.

1. By direct attack against antigens

2. By activation of complement system

Because of prevalent nature of the antibodies and

the multiple antigenic sites on most invading agents
antibodies incultivate the invading agents in one of the
following ways invading agents in one of the following ways.
1. Agglutination

2. Precipitation

3. Neutralization

4. Lysis
Innate or Inborn or natural immunity :

Ability of human body to resist the occurrence of

disease by general processes rather than by specific
immune system is called innate immunity, it is not
acquired from previous attack of disease. It forms non-
specific defence mechanism of human body. It is similar for
most type of the infections. Thus non-specific mechanism
includes lines of defence.

First line of defense: It includes skin, lysozyme,

complement system, interferon.

1. Skin or integumental innate defenses :

a) Mucus: When the fish is attacked by the bacteria the

mucus is continually being produced and sloughed from
the integumental surface thus physically trapping and
preventing the bacteria from attaching to the epithelium
and having an opportunity to invade the fish’s tissue.
Mucus contains many substances which have been
identified from mucus secretions of number of fish species.

Anti-bacterial peptides - e.g. pleurocidin :

The gene encoding anti-bacterial peptide pleurocidin

has been done from the winter flounder and shown to be
predominantly expressed in skin and intestinal tissue and
gene is first expressed as early as 13 days post hatch.

Thus these peptide may provide an important line of

defence before development of specific immune response in
larval fish.

Synthetic pleurocidin has recently been shown to

protect cholo salman from infection by V. anguillarum.

Proteases : Trypsin-like proteases and cathepsin proteases

have been found in skin mucus of a number of fish species.

Lectins : These are a group of proteins with different

specificities for binding carbohydrates. They have been
found in salmon eggs.

These lectins are Ca2+ dependent and can

agglutinate a number of fish bacterial pathogens.

It is a possibility :

Mucus Ò Lectins Ò Carbohydrate binding and attachment

to surface - blocking of pathogen invasion

Lysozyme : This enzyme can attack the peptidoglycan layer

of bacterial cell walls causing them to lyse. Lysozyme has
been found in fish mucus, serum and ova. Peritoneal
macrophages and blood neutrophils contain lysozyme and
latter are thought to be the source of serum lysozyme. Fish
lysozyme occurs in 2 forms and one of these appear to be
much more bactericidal than lysozyme of higher
vertebrates.
Complement system :

If bacteria are successful in crossing the

integumental defense then there are number of plasma
proteins which may prevent further infiltration. Most
important serum defense factor is the complement system
because of its activating effect or effector mechanism on
cellular defenses.

Binding of antibody to antigen often triggers the

complement system through so called classical pathways
and it includes.

C1 to C9 proteins are includes :

C1 exists in blood serum it contains 6 molecules of

C1q and 2 molecules of C1r and C1s types of proteins.

1. Constant region of 1gM contain binding site for C1q.

It combines with C1q activates C1s and C1r.

2. Activated C1 is cleaved into C4 and C2. There is

complex formation called C4a which is known as C3
convertase. C4a causes activation of mast cells and
basophils.
3. C3 is cut into C3b and C3a by C3 convertase C3
blinds to glycoproteins scattered all over.
Macrophages and neutrophils have receptor for C 3b
and bind the C3b coated cell. This effect qualifies
C3b as an opsonin to enhance phagocytosis. The
process by which particulate antigens are rendered
more susceptible to phagocytosis is called
opsonization.
 Some of C3b binds directly to C42 producing a
trimolecular complex C423b which act of C5, it is called C5
convertase.

Cleavage of C5 by C5 convertase initiates the

cleavage of set of complement proteins that make up
membrane attack complex.

C5 cleaves into C5a and C5b.

C5a is released into surroundings and act as chemotoactic

attractant for neutrophils.

C5b serves anchor for assembly of single molecule of each

C6, C7 and C8.

The resulting complex C5b678 guides polymerization

of 18 molecules of C9 into a tube inserted into lipid bilayer
of plasma membrane. This tube forms a channel allowing
passage of ions and small molecules. Water enters the cell
by osmosis and the cell lyses.

The second line of defense comprises of actions of

leukocytes :

Basophils are found in some marine species.

Granulocytes are phagocytic, involved in combating disease
and many increase in number when fish is infected by
bacteria. Lymphocytes include phagocytic macrophages,
plasma cells.

Leucocytes are formed in kidneys of many teleosts

but in elasmobranchs there is a special organ (organ of
leydig) that assumes this function.
 Major lymphoid organs in teleost fish are thymus,
kidney and spleen. Thymus is composed of developing
lymphocytes.

Kidney is main antibody producing organ. It contains

generalized haemopoietic tissues rich in lymphocytes. It is
also a filtration organ containing many macrophages which
phagocytase antigen.

Spleen contains fewer haemopaietic and lymphoid

cells than kidney, being composed mainly of blood held in
sinuses. It contains specialized capillary walls termed
ellipsoids which one composed of reticulin fibres and
macrophages. Macrophages are highly phagocytic. While
reticulin fibre network is specialized for trapping complexes
of Ag-Ab.

Second line of defense is also called macrophage

system and is non-specific internal defense.

Especially neutrophils of blood have ability to come

out of blood capillaries by amoeboid movement called
Diapedesis and engulf and digest many of bacteria which
cause infection. Process of engulfing bacteria by cells of
macrophage systems is called phagocytosis.

Mechanism of phagocytosis :

1. Neutrophil first attaches itself to the particle to be

phagocytized.

2. It out pseudopodia around the particle to form a cup.

3. Pseudopodia fuse and ingest the particle in a food

vacuole called phagosome. In this way, a neutrophil
can phagocytize 5 to 20 bacteria.

4. Phagosome and primary lysosomes like proteolytic

enzymes and large amount of lipase fuse to form
 secondary lysosome or digestive granule. These
enzymes digest bacteria.

5. Diffusible material diffuses in cytoplasm while

undigested material remains in the phagosome
which is now called residual body. It is expelled out
of cell by exocytosis.

Macrophages :

These are large amoeboid phagocytic cells which are

found in most of connective tissue, spleen, reticulo
endothelium, liver. They engulf particular material
including microbes, tissue debris, dead cells etc., act like
scavenger. Along with cells of lymphocytes they form
lymphoreticular or reticuloendothelial system. Macrophage
may fuse to form a large multinucleate giant cell to capture
a large sized foreign body.

Macrophage differ from leucocytes. Macrophages

have much greater ability to engulf inorganic particulate
materials and are also responsible for phagocytosis but
they can produce antibodies.

Inflammatory response :

Leucocytes and macrophages which form second line

of defence always operate through inflammatory response.
According to this response when micro-organisms like
bacteria, viruses etc. enter the body tissue through some
injury, these produce some toxic substances which kill
tissue cells. Damaged cells produce histamine which causes
inflammation characterized by dilation of capillaries and
small blood vessel surrounding the injury increasing the
blood flow injured tissues. The infected area becomes red,
warm and swollen increase in permeability of the capillary
wall. Plasma leaks into the tissue space diluting the toxins
secreted by bacteria. The phagocytes i.e. neutrophils and
macrophages show chemotactic response and are attracted
by the chemicals released from inflammed area like
histamine, reaction products of complement system and
blood clotting system and lymphokines of sensitized T cells.

Immune system shows either of two responses cell

mediated and humerol immune response.

Cell mediated immune response is the result of T cell

whereas B cells contribute to the humoral immune
response.

B cell and T cells are Z kinds of lymphocytes.

Mode of action of B lymphocytes :

There are thousands of antigen specific B cells in the

body. Though B cells have capacity to produce specific
antibody even during development and differentiation, but
these produce antibody only when stimulated by antigen.
Once B lymphocytes are stimulated by an antigen by
contact, they synthesize RNA, divide rapidly and
differentiate into rough endoplasmic reticulum rich
histologically distinctive plasma cells. A group of plasma
cells is known as clone which produce antibodies at a very
high rate. These antibodies circulate in the lymphs to fight
the antigen so forming humoral immune system. B
lymphocytes are short lived and they are replaced by new
cells after every few days. Some of the lymphoblasts do not
form plasma cells but form moderate number of new B-
lymphocyte called memory cells. Memory cells circulate
throughout the body, remain in lymphoid tissues but
remain inactive. These cause much more potent antibody
response when exposed to same response called secondary
response. This explains the fact that vaccination is usually
accomplished by injecting antigens in multiple doses with
some period in between the injections.
Mode of action of T-cells :

T cells are also antigen specific as each T-cell

recognizes a specific antigen and different types of T-cells
are stimulated by different types of antigens.

Immunologically competent T lymphocytes divides

rapidly to form clone of T-cells called T-lymphoblasts. These
activated T cells are released into lymph, then pass into
blood and distributed throughout the body though
morphologically similar but they are divided into 3 classes
on the basis of their function.

Helper-T cells : Most numerous. These recognize and bind

to class II MHC molecule present on the surface of antigen
and produce a variety of soluble factors called cytokinin
which include lymphokines, inter leukins, monokines,
interferons which act in a different ways on immune
system.

Killer or cytotoxic T cells :

Killer T cells represent other major type of

lymphocytes which migrate to the site of infection and
release lymphokines (which attract macrophages by
chemotasis), lymphotoxins (which kill foreign cells by
secreting cytokyines which damage cell membrane of
microbes) and interferons which in activate viruses. So
phagocytic activity is highly enhanced at the site of
infection.

Suppressor T-cells :

These suppress the whole immune system to protect

the own body cells from its attack. These are capable of
suppressing the functions of both cytotoxic and helper T-
cells as these regulate activities of other cells, checking
them from causing excessive immune reactions. Helper T-
cells activate the suppressor T cells, while suppressor T
cells act as a negative feed back controller of helper T cells.
So both helper T cells and suppressor T cells are called as
regulatory T cells.

After discussing about all the components of fish

immune system we can conclude that it is not much
different from immune system of other vertebrates. It has
only one kind of immunoglobulin i.e. IgM unlike vertebrates
which have kinds of antibodies and one of its lysozyme is
more bactericidal in comparison to mammalian lysozymes.

Pawan Kumar, Mukta Sharma and Ravi Kant Verma

 Status of Fish Vaccines

Microbial diseases are a major problem for aquaculture

all over the world. From both health and economic
perspectives, it is far more beneficial to prevent a
disease from happening then it is to treat the disease
after it strikes. Until recently only treatment for many
viral diseases in the aquaculture industry has been the
destruction of injected animals. But now disease
management strategies based on vaccines are being
increasingly explored (Karunasagar et al., 1999).

What is vaccine?

A vaccine is a small amount of weakened or killed

disease producing microbes that are introduced into the
body of organism to prevent the occurrence of the
disease they produce.

The process of administering vaccines is known as

vaccination.

Characteristics of an ideal vaccine.

1 The ideal vaccine to be used for active immunization

should be cheap, stable and adaptable to mass vaccination.

2. It should give prolonged strong immunity without

adverse side effects and ideally should stimulate an
immune response distinguishable from that due to natural
infection so that vaccination does not interfv3ere with
diagnosis.
3. It should be free from adverse side effects.

How to vaccines work?

Through vaccine immunogenic but non-virulent antigen

is introduced in the body of organisms. These antigens
are recognized by specific lymphocytes. Lymphocytes are
of 2 types T cell and B cell . T lymphocytes are produced
from Thymus and B lymphocytes from bone marrow in
mammals and probably kidney in teleost fish. On
exposure to antigen, B cells differentiate into plasma
cells which produce antibody specific to the simulating
antigen or B cells they differentiate into memory cells
which become active in case of subsequent natural
infection or booster vaccination.

T cells – are of 3 types i.e. helper cells, memory and

suppressor cells. On first exposure to antigen helper T cells
cooperate with B cells and produce memory T cells. Memory
T cells cooperate with B cells on further infection.
Suppressor T cells.

The commonly used vaccine in aquaculture are –

1. Attenuated vaccine – In these vaccines living organism
is used but they

are treated in such a way that they loose their disease

producing ability.

Attenuation can be done by –

Adapting organisms to unusual environmental conditions

so that they loosed the ability to replicate uncontrollably in
their usual hose. (By growing in abnormal culture
conditions, i.e. growing on cell/issue which are not usually
infected by organism)/ Adopting the virus to growth at
slightly lower temp. than normal).

Advantages- In general vaccines containing living organisms

are effective and give prolonged, strong immunity. In fish
they can be administered by water borne route, which is a
cheap and effective method. Also few inoculating doses are
required and adjuvant need not be employed.
Disadvantages -They are expensive and difficult to
produce. There exists the possibility that the attenuated
vaccine might cause disease due to residual virulence (if not
fully attenuated) or due to reversion to a more virulent

These vaccines are under development for.

Aeromonas .hydrophila, Renibacterium
salmoninarum

Viral hemorrhagic septicemia

2. Killed vaccines –

To avoid the disadvantages to live vaccines, it is

common to sue organisms that have been killed or
inactivated. In such cases the dead organism should be
as antigenically similar to the living organisms as
possible. Commonly used inactivating agents includes –
formaldehyde

ethylene oxide, Ethylenimine

Advantages
They contain no residual virulence so cannot cause
disease.

Disadvantage
They are less effective than live vaccines. They cannot be
administrated effectively by water borne routes (Leong et
al).

Y.ruckeri, bacterin was the first commercially produce9d

fish vaccine. It was a formalin killed whole cell product.

Most of the commercially available vaccines (killed vaccines)

are against:
- Edwardsiella ictaluri
- Infectious pancreatic necrosis virus
- Viral hamarrhagic septicaemia
- Infectious haematopoietic necrosis virus
Conventional vaccines has many disadvantages and there is
a need for improvement. Several new approaches are being
studied in attempt to make vaccines more effective,
cheaper, and safer. These are –

Purified antigen vaccine

These vaccines are based on purified antigens
isolated from the concerned pathogen.

Advantages –
The risk of pathogenicity is avoided as they do not
contain the organism.

Disadvantage:

 The cost of production is high. (due to steps

involved in purification and vaccine preparation).
 Many of the isolated antigens are poorly
immunogenic.

Recombinant vaccines –:
A recombinant vaccine contain either a protein or a gene
encoding a protein of a pathogen origin. The vaccines
 are produced using recombinant DNA technology
(RDT).These are of mainly two types

1. Recombinant protein vaccines. They are also called

subunit vaccines. These are of further two types.

Preparation –
First step is to identify a protein that is both
immunogenic and critical

for the pathogen.

Gene encoding this protein is then identified and

isolated.

The gene is integrated into a suitable expression vector

and introduced into a suitable host where it expresses
the protein in large quantities.

The protein is then isolated and purified from the host

cells and used in the preparation of vaccines.
Whole protein molecules

In these vaccines complete immunogenic protein is

produced in genetically engineered microbes.

Polypeptide vaccine
The immunogenic property is usually confined to a
small portion of the protein molecule. Therefore, whole
protein molecule is not necessary for immunogenicity.

e.g. Cholera enterotoxin produced by vibrio cholerae

consists 3 polypeptide viz. A12, A2, B polypeptide. The
A1, A2 polypeptide are toxic but B is non-toxic and
imunogenic. The gene encoding B polypeptide is cloned
and recombinant are produced.
Advantages
They are safe as they do not involve whole organism.

Disadvantages-
Cost of production is high

They have to be stored at low temperature otherwise

protein can get denatured

DNA VACCINE
In these vaccines genes encoding the relevant
immunogenic protein is isolated, cloned and then integrated
into a suitable expression vectors. This preparation is
introduced into the individual to be immunized.

ADVANTAGES
1- Purification and preparation of DNA for vaccine is easier
cheaper

2. They are safer and more specific because of high purity

METHOD OF ADMINSTERING VACCINE

1. INJECTION / INOCULATION
This is the most effective way of vaccinating fish. In this
method fish are kept in a large holding tank from where
they are transferred to anesthetic tank. The common
anesthetics in general use are MS222 and benzocaine.
Anesthesia is necessary not only to facilitate handling
but also to avoid excessive stress. They are then passed
to injection table where vaccine is injected
intraperitoneal.

ADVANTAGES –
 It is the most potent immunization route.
 It allows use of adjuvants.
 It is a very cost effect method for large fish.
DISADVANTAGES

 It is useful only in intensive aquaculture

 It is labour intensive method.
 It is a highly stressful method.
 It involves use of anesthesia.
 Fish weigh must be >15 gm for the success of this
method.
2. ORAL VACCINATION
In this method vaccine is added directly in to the feed.

ADVANTAGES
 It is the only effective method for extensive
aquaculture.
 It is a non stressful method.
 It allows mass vaccination of fish of any
size
 No extra labour cost is involved in this
method.

DISADVANTAGES
 This method has poor potency
 Large amount of vaccine is required in this
process.
 It is suitable only for fish fed on artificial
diet

IMMERSION VACINATION
In this technique outer surface of the fish is directly
exposed to the vaccine. Immersion vaccination method
are of mainly three types differing in the dilution of
vaccine used and exposure time required for
vaccination as shown in the following table.
 Route Dilution Exposure time

Immersion dip 1:3 /1:10/1:100 5-30 s

Immersion bath 1 : 500 / 1: 5000 1hr or longer

(least stressful)

Spray 1 : 3 2-5 s
/1:10/1:100

ADVANTAGES
 It allows mass vaccination of small fish (< 5 g)
 It is the most cost effective for fish < 10 gm
 It is comparatively less stressful method.
DISADVANTAGES

- Only for intensive aquaculture

Potency not a fish as injection route

The ability of fish to develop immunity to a disease by

vaccination is dependent on two Kinds of factors –
1- Intrinsic factors or fish related factors:

(1) Size of the fish

Most studies indicate that the size of the fish at the
time of immunization is critical for the development of
optimum response. In general, large and thus more
mature the fish at the time of immunization, the greater
is the level and duration of protective immunity,
e.g. Protection against ERM and vibriosis increased with
size when fish are vaccinated duration of protection
(solenoid)
no immunity 0.5 g fish
 120 days 1 g fish
180 days 2 g fish
1 year 4 g fish
( Johnson et al.,1982)

Extrinsic Factors
Temperature – Temperature which are low with in
the normal physiological range of a particular species of
fish can depress the immune response (e.g. less than 6 0C in
rainbow 120C for carp) e.g. In carp which were
acclimated to 12, 16, 18, 20 or 240C for 2 weeks before
vaccination, the primary antibody response was delayed by
9 days at 25C and 49 days in 120C. (Rijkers etal.,1980)

In case of vaccination at temperature below 180C, there was

no induction of immune memory.

Note - low temperature depress the immune response of t

dependent antigen. It is advisable to fish farmers to
artificially raise the temperature.

Diet : In order to have efficient immune response to

vaccination, fish should be healthy and in good condition.
Therefore, it is necessary to provide balanced diet for
efficient immune response. Very few studies have been
made regarding the effect of nutrient to immune response

e.g. –Effect of vitamin C and vitamin. E.

The antibody response to E. ictaturi antigens was antigens

was lower in channel cut fish given vit C deficient diets
than in those given normal dose (30-300 mg Vit.C/kg diet)
Mega dose amount (3000 mg Vitamin c/kg diet)
significantly enhanced antibody responses still further ( Li
etal.,1985)
Rainbow trout fed Vitamin E deficient diet for 12-17
week has reduced antibody response compared with control
fish fed at a diet containing 40 Iu Vit. E/100 g diet.
(Blazer et al., 1984)

Pollutant:

Various pollutants are known to suppress the immune

response of fish or increase disease susceptibility.

(a) The pollutant may inhibit the uptake of antigen from

bath immunization by affecting gill permeability.(eg.
Phenol).
(b) Pollutant induce persistent stress in fish causing
suppression of the immune response.
(c) They directly interfere with cells or their products
involved in defense (heavy metals).
Pollutant Concentratio Fish Decreased response
n
Immune To
(ppm)

Cd Sub lethal Carp, brown CMI suppressed

trout viruses
Cr

Cu 0.01 –0.03 Coho salmon Vibrio

Cu 0.03 – 0.06 Eel, carp Vibriosis

Sn 0.3 Zebra fish Bacteria

Phenol 12.5 Carp Aeromonas

Phenol 10 Rainbow trout Yersinia suckeri

 O’Neill, J.G., 1981; Rijkers et al., 1980; Zeeman et al., 1981

Antibiotics
Antibiotics are known to affect the immune response
in mammals but the effects are variable depending on
the antibiotic-Little is unknown about this subject in
fish e.g. concern has been expressed about immune
suppressive effect of oxytetracyline OTC in carp and
rainbow

Status of vaccine development against various

diseases –

1. Vaccination against vibriosis.

Vibriosis is a bacterial disease of salt water and

migratory fish.

Vaccine developed –

Several commercial vaccines are available; most contain

V. anguillarum serotype 1 and 2. Polyvalent vaccines
with other antigen such as V.salmonicida ,
A.salmonicida. are also available.(Toranzo,1997)

2. Vaccination against Enteric RM (ERM)

Potential host – Rainbow trout, salmonid sp.

Vaccine developed –
Oral vaccine is available which is prepared from phenol
killed bacteria.(Stevenson,1997)
3. Furunculosis Vaccine –

Bacterial disease of salmonid fish caused by

Aeromonoas salmonicida.
Vaccine developed –
First attempt to vaccinate fish was against
furunculosis by using chloroform killed Aeromonas
salmonid cells throat trout.

Commercial vaccines are available they are effective will

oil based adjuvants. Recombinant vaccines are under
development phase (Duff, 1942; Ellis, 1997).
4. Vaccination against Edwardsiella tarda-

first reported as causative agent of disease in Japanese eel

and later in variety of cultured fresh water and sea water
fish such as gold fish, tilapia, Rainbow trout etc.

Vaccines developed –
Two types of vaccines are used-bacterins and bacterial
extracts.No commercial vaccines are available.(Song et
al.,1981)

5. Vaccination against Bacterial kidney disease (BKD)

Disease is caused by Renibacterium salmonarum. It causes
serious mortality in salmonid fish.

Effective vaccine is yet to be reported. (Kaattari,1997)

6. Vaccination against Edwardsiella ictaluri-

It causes enteric septicemia of channel catfish (ESC)

Vaccine developed –
Orally delivered killed vaccine is licensed. (Thune
etal.,1997)

7.Vaccination against Infectious Pancreatic Necrosis

(IPN)

It is a viral disease in salmonid fish.

Vaccine developed –
Inactivated viral vaccines give inconsistent results.
Recombinant vaccines have also been reported.
Injectable vaccine based on viral protein VP2 has
been licensed. (Christie et al., 1997)

8. Vaccination against Viral hemorrhagic septicemia

It is caused by rhabdovirus in grayling, white fish etc.

Vaccine developed –
Infection of killed viral vaccine gives protection.
Attenuated and recombinant vaccines are under
development phase (Lorenzon et al., 1997).

Manisha Gupta and Pawan Kumar

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