Blood and

tissue protozoa
(con’t)

DR.MEHRU NISHA
mehrunisha@unikl.edu.my
 Leishmania
• Species Leishmania is widely distributed in
nature
• Example of Leishmania species:-
i) Leishmania donovani Old World Leishmaniasis
(bite by female sandflies of
ii) Leishmania major the genus Phlebotomus)

iii) Leishmania chagasi New World Leishmaniasis

(bite by sandflies of the
iv) Leishmania peruviana genera Lutzomyia)

Distribution mainly in Indian subcontinent,

Tropical South America, Ethiopia and
Kenya
 The Parasite
• Phylum Sarcomastigophora

• Order Kinetoplastida

• Family Trypanosomatidae

• Genus Leishmania
 Morphology
• Promasitogte • Amastigote
– Insect – Mammalian stage
– Motile – Non-motile
– Midgut – Intracellular

Amastigotes (*) of Leishmania

donovani in the cells of a spleen.  The
individual amastigotes measure
approximately 1 µm in diameter. 
 Leishmaniasis in Humans

Parasite Disease

L.donovani Visceral leishmaniasis

(kala-azar,dumdum, fever

L.tropica Cutenous leishmaniasis

(Delhi boil)

L.braziliensis Mucocutaneous leishmaniasis

(American leishmaniasis)
 Life Cycle

1. Leishmaniasis is transmitted by the bite of infected female phlebotomine

sandflies.
2. The sandflies inject the infective stage (i.e., promastigotes) from their proboscis
during blood meals .
3. Promastigotes that reach the puncture wound are phagocytized by
macrophages and other types of mononuclear phagocytic cells.
4. Progmastigotes transform in these cells into the tissue stage of the parasite
(i.e., amastigotes) , which multiply by simple division and proceed to infect
other mononuclear phagocytic cells .
5. Parasite, host, and other factors affect whether the infection becomes
symptomatic and whether cutaneous or visceral leishmaniasis results.
6. Sandflies become infected by ingesting infected cells during blood meals.
7. In sandflies, amastigotes transform into promastigotes, develop in the gut (in
the hindgut for leishmanial organisms in the Viannia subgenus; in the midgut
for organisms in the Leishmania subgenus), and migrate to the proboscis .
 Mammalian Hosts
• Rodents • Sloths
• Gerbils • Primates
• Hyraxes • Dogs
• Bats • Foxes
• Porcupines • Anteaters
• Opossums • Human
 Pathogenicity
1. Visceral leishmaniasis/ kala-azar
 occurs in India like in Bihar, West bengal,Africa
eastern districts of Uttar Pradesh,
 incubation period 10 days/3-6 months
 parasites spreads from site of inoculation to
multiply in reticuloendothelial cells
 especially in spleen, liver , lymph nodes and
borne marrow
 caused by Lutzomyia sandflies
 Disease manifestation
o Fever
o Malaise
o Headache
o Progressive enlargement of spleen
o Liver enlargement
o Lymph nodes
o Anemia
o Emaciation
• Profile view of a
teenage boy suffering
from visceral
leishmaniasis. The boy
exhibits splenomegaly,
distended abdomen
and severe muscle
wasting. 
• A 12-year-old boy
suffering from visceral
leishmaniasis. The
boy exhibits
splenomegaly and
severe muscle
wasting.
• Jaundiced hands of
a visceral
leishmaniasis
patient. 
• Enlarged spleen and
liver in an autopsy of
an infant dying of
visceral leishmaniasis.
Cutaneous leishmaniasis of the face. 
A cutaneous leishmaniasis lesion on
the arm.
 Laboratory Diagnostic
Non –Specific Laboratory test Parasitology Diagnostic

Blood count Peripheral blood film

-erythrocytes will be decreased

Haemoglobin estimation Needle biopsy/aspiration

-it reveals anaemia

Estimation of serum proteins Culture

-it reveals raised serum proteins -Hockmeyer’s medium

 Treatment
Good nursing
Diet
Antibiotics
Pentavalent antimony
Pentamidine

New drugs - New delivery

 Control
• Vector control
• Reservoir control
• Treatment of active cases
 Trypanosomes
 Tryoanosomes are haemoflagellates that live
in the blood and tissues of their human hosts
 Trypanosoma brucei brucei,
T.brucei.gambiense & T.brucei. rhodesiense
 Trypanosoma brucei gambiense
Laboratory Diagnosis 7
 Thick and thin blood smear 8
Treatment, Prevention and Control
 Aspirations from lymph
- Suramin (anti-parasite drugs)
nodes & concentrated spinal
- Melarsoprol
fluid
Clinical syndromes: 6 - Use of insecticides, wearing
 Serological test-ELISA,
- African sleeping protective clothing, bed
Immunofluorescence
sickness netting, insect repellent
- Fever
- Lymph node
enlargement
- Myalgia (muscle
pain) The infective stage of the
- Chronic disease organism is trypomastigote,
1
progresses to present in salivary glands
CNS system, of transmitting tsetse flies
involvement with
lethargy, tremors,
mental
The organism in this stage has 2
retardation and
a flagellum & an undulating
general
membrane running full length
deterioration
of the body

5
T.B gambiense is
4
limited to tropical West Tsetse flies become infective 4 3
The trypomastigotes enter the
& Central Africa-river to 6 weeks after feeding on
wound created by fly bite & find the
banks blood from a diseased patient
way into blood, lymph, and finally
invading CNS system
 Trypanosoma brucei (sleeping sickness)
• Protozoan hemoflagellates belonging to the complex
Trypanosoma brucei.
• Two subspecies that are morphologically indistinguishable
cause distinct disease patterns in humans: T. b. gambiense
causes West African sleeping sickness and T. b. rhodesiense
causes East African sleeping sickness.
 Trypanosoma cruzi (chagas disease)

• Also known as kissing bugs because they frequently bite

people around the mouth area

• This organism in the feces of the bug enter the wound and
penetration happens when patients rubs or scratches the
irritated site

• The trypomastigotes migrate to other tissues (e.g cardiac

muscle, liver, brain)—and become the smaller, oval,
intracellular amastigote form

• https
://www.isglobal.org/en_GB/-/enfermedad-de-chagas-ciclo-del-parasito-en
-humanos
 Epidemiology T.Cruzi occurs in North,
Central and South America

Treatment, Prevention and

Control Clinical Syndromes

- Nifurtimox (but little less effective - Chagas disease

- Chagas disease can be
against amastigotes
asymptomatic, acute, chronic
- Alternate - One of the earliest sign is the
agents include
allopurinol and benzimidazole development of swollen
eyes/nose area (chagoma)
- Bug control, eradication of nests, - Acute infection causes fever,
use of dichlorodiphenyl- chills, malaise, fatigue
- Death can occur after long
trichloroethane
term of acute attack due to
Laboratory diagnosis parasite proliferation at heart,
-T.Cruzi can be demonstrated in thick liver, spleen, brain and lymph
and thin blood films nodes
Biopsy of lymph nodes, liver, spleen or
bone marrow
-Polymerase chain reaction (PCR)
 Naegleria fowleri
• Naegleria fowleri trophozoites are found in cerebrospinal fluid
(CSF) and tissue, while flagellated forms are occasionally
found in CSF.
• If the environment is not conducive to continued feeding and
growth (like cold temperatures, food becomes scarce) the
ameba or flagellate will form a cyst.
• The cyst form is spherical and about 7-15 µm in diameter. It
has a smooth, single-layered wall with a single nucleus.
• Cysts are environmentally resistant in order to increase the
chances of survival until better environmental conditions
occur
Naegleria fowleri has 3 stages in its life cycle:

(1), cyst (2) trophozoite, and (3) flagellate. The

only infective stage of the ameba is the
trophozoite. Trophozoites are 10-35 µm long
with a granular appearance and a single
nucleus.

(4) The trophozoites replicate by binary division

during which the nuclear membrane
remains intact (a process called promitosis)

(5) Trophozoites infect humans or animals by

penetrating the nasal tissue and migrating
to the brain

(6) via the olfactory nerves causing primary

amebic meningoencephalitis (PAM).

Trophozoites can turn into a temporary, non-

feeding, flagellated stage (10-16 µm in length)
when stimulated by adverse environmental
changes such as a reduced food source. They
revert back to the trophozoite stage when
favorable conditions return.
 ns

pl
ic Primary Amebic
at
io

m
Co
Meningoencephalitis (PAM)
o 1-14 days incubation period
o symptoms usually within a few days after swimming in
warm still waters
o infection believed to be introduced through nasal cavity and
olfactory bulbs
o symptoms include headache, lethargy, disorientation, coma
o rapid clinical course, death in 4-5 days after onset of
symptoms
o trophozoites can be detected in spinal fluid, but diagnosis is
usually at autopsy
o known survivors treated with Amphotericin B
Primary Amebic Meningoencephalitis
(PAM)
 DIAGNOSIS
DIRECT VISUALIZATION
 by microscopic examination of fresh, unfrozen, unrefrigerated cerebrospinal
fluid (CSF). Samples cannot be frozen or refrigerated because cold temperatures
kill the ameba.  can also be stained with a variety of stains, such as Giemsa-
Wright or a modified trichrome stain, for identification

POLYMERASE CHAIN REACTION (PCR)

Specific molecular tools can amplify DNA from the amebae in CSF or tissue to
specifically identify if the amebae are present.

CULTURE
 The amebae can be grown in culture to increase the likelihood of detecting the
ameba by direct visualization or PCR.

IMMUNOHISTOCHEMICAL STAINING
 Treatment
• Antiamoebic drugs and antibacterial
antibodies are ineffective.
• Amphotericin B has been used for treatment
and clinically effective.
 PREVENTION
• To refrain from water-related activities in warm freshwater
These actions could include:
– You cannot get infected from contaminated drinking water.
You can only be infected when contaminated water goes up
into your nose.
– Hold your nose shut, use nose clips, or keep your head above
water when taking part in water-related activities
– Avoid water-related activities in warm freshwater during
periods of high water temperature.
– Avoid digging in, or stirring up, the sediment while taking part
in water-related activities in shallow, warm freshwater areas.
THANK YOU