MODULE Leishmaniasis
Microbiology
42
Notes
LEISHMANIASIS
42.1 INTRODUCTION
Sir William Leishman and Charles Donovan demonstrated the parasite in
patients from Calcutta in year 1903. The Genus was so named by Sir Ronald
Ross-Leishmania Donovani. It was first cultured by Rogers in 1904.
Geographic distribution-It is mainly seen in South and South –East Asia, China,
Sudan tropical Africa South America. In India the regions mainly affected are
– Bengal, Bihar, and Eastern Uttar Pradesh
OBJECTIVES
After reading this lesson, you will be able to:
z describe the morphology of Leishmaniasis
z explain the life cycle of Leishmaniasis
z discuss the pathogenecity of Leishmaniasis
z explain the laboratory diagnosis of Leishmaniasis
42.2 HABITAT
Amastigote forms are seen in human infections and are seen mainly in the cells
of reticuloendothelial system located in liver, spleen, bone marrow, and
peripheral blood
Promastigote forms are seen in the gut of sand fly Phlebotomus argentipes. It
is also seen when grown in laboratory on artificial culture media.
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Leishmaniasis MODULE
Microbiology
42.3 MORPHOLOGY
(a) Amastigote: These are non motile and round to oval 2-4 µm long.
Notes
Fig. 42.1
Nucleus is round to oval. The nucleus is red and the kinetoplast is bright
red on Leishman stain
A clear unstained space is present alongside the axoneme called the
vacuole. The parasite has the blepharoplast and axoneme. Axoneme, arises
from the blepheroplast and extends to the margin of the parasite. Amastigote
form also has a parabasal body.
(b) Promastigote: It is a spindle shaped structure measuring 15-20 um by
1-2 um. It has a flagellum arising from the axoneme and coming out of
the anterior end. There is a blepheroplast and a vacuole in the anterior end.
Nucleus is round to oval and central in location. The nucleus is red and
the kinetoplast is bright red on Leishman stain
Fig. 42.2
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MODULE Leishmaniasis
Microbiology
42.4 LIFE CYCLE
The parasite is transmitted amongst humans by the bite of infected Sand Fly-
Phlebotomus and Lutzomiya Transmission can also occur through infected
blood transfusion.
After infected blood meal is taken by the vector sand fly Phlebotomus argentipes,
the aflagellates develop into flagellate promastigote form in the gut of the insect
Notes in 8-20 days. The infective sand fly transmits the disease by biting man. Due
to partial/complete blockage of mouth parts Parasites is lodged at site of bite
when it ingests blood.
The promastigotes penetrate the host macrophage and are converted to
amastigote forms. They multiply and rupture the macrophages. There is
ingestion of amastigote by other macrophages.
Incubation period: Usually 3-6 months, but can extend up to 1-2 years
Fig. 42.3
376 MICROBIOLOGY
Leishmaniasis MODULE
Microbiology
INTEXT QUESTIONS 42.1
1. Amastigote forms are seen in ........................
2. Promastigote forms are seen in ........................
3. Unstained space present alongside axoneme called ........................ Notes
4. Promastigote is transmitted by ........................
5. Kala azar is caused by ........................
42.5 PATHOGENECITY
Leishmania donovani causes a visceral disease called Kala azar. It is a parasite
of the reticulo endothelial (RE) system and affects the organs containing the
reticuloendothelial system like the bone marrow, liver and spleen. There is
hepatosplenomegaly. The patient gets intermittant fever. There is associated
anaemia, cachexia, loss of weight. There is dry skin, brittle hair and
pigmentation of skin. There may also be diarrhoea, dysentery. Oedema is seen
due to hypoalbuminemia.
Leishmania tropica causes Oriental sore (cutaneous leishmaniasis) On the skin
which becomes dry there may appear solitary or multiple ulcerating papules.
Healing occurs with scarification. Post kala azar dermal leishmaniasis
Leishmania brazilensis- Espundia (muco cutaneous leishmaniasis)
42.6 LABORATORY DIAGNOSIS
Demonstration of L-D bodies: The aetiological diagnosis is established by
demonstrating the parasite in the patient specimen. The amastigote form of the
parasite is seen in human infections. Parasite demonstration is done on the
following specimen after staining the smears with Romanowsky stains.like
Giemsa, Lieshman stain.
z splenic aspirate
z bone marrow smears
z thick blood film
z enriched blood leucocyte fraction
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MODULE Leishmaniasis
Microbiology
Notes
Fig. 42.4
Detection of specific antibodies: Antibodies can be detected in immunocompetant
individuals in 99% of cases. ELISA, CIEP, DOT ELISA based kits are available
for the same. In immunocompromised individuals like HIV patients with AIDS
the 40-60% cases may be seronegative.
Culture on monophasic/biphasic media: The promastigote form of the
parasite is seen on artificial culture media. The patient specimen can be cultured
on the following media
z N N N media (Novy,MacNeal Nicolle)
z Brain heart infusion agar medium
z Schneider’s medium
Fig. 42.5
378 MICROBIOLOGY
Leishmaniasis MODULE
Detection of antigen: Antigen detection kits based on ELISA, IFA are available Microbiology
and can be used for establishing the diagnosis.
Non-specific tests
z progressive neutropenia
z relative lymphoctosis,monocytosis
z reversal of A:G ratio
Notes
z Napiers Aldehyde test
z Chopras Antimony test
Newer tests: PCR based detection can be done in laboratories having molecular
biology facility.
Isoenzyme typing is one of the typing method which may be done in research
laboratories.
INTEXT QUESTIONS 42.2
1. Kala azar (a) Leishmania Donovani
2. Oriental sore (b) Leishmania brazilensis
3. Espundia (c) Leishmania Donovani
WHAT HAVE YOU LEARNT
z Amastigote forms are seen in human infections and are seen mainly in the
cells of reticuloendothelial system located in liver, spleen, bone marrow,
and peripheral blood
z Promastigote forms are seen in the gut of sand fly Phlebotomus argentipes.
It is also seen when grown in laboratory on artificial culture media.
z The parasite is transmitted amongst humans by the bite of infected Sand
Fly-Phlebotomus and Lutzomiya Transmission can also occur through
infected blood transfusion.
z Incubation period is usually 3-6 months, but can extend up to 1-2 years
z Leishmania donovani causes a visceral disease called Kala azar, Leishmania
tropica causes Oriental sore and Leishmania brazilensis causes Espundia
z The aetiological diagnosis is established by demonstrating the parasite in
the patient specimen
z Parasite demonstration is done after staining the smears with Romanowsky
stains.like Giemsa, Lieshman stain
MICROBIOLOGY 379
MODULE Leishmaniasis
Microbiology z Antibodies can be detected in immunocompetant individuals, ELISA, CIEP,
DOT ELISA based kits are available for the same.
z The promastigote form of the parasite is seen on artificial culture media.
The patient specimen can be cultured on the following media N N N media
(Novy,MacNeal Nicolle), Brain heart infusion agar medium, Schneider’s
medium,
z Antigen detection kits based on ELISA, IFA are available and can be used
Notes
for establishing the diagnosis.
z PCR based detection can be done in laboratories having molecular biology
facility and Isoenzyme typing is one of the typing method which may be
done in research laboratories.
TERMINAL QUESTIONS
1. Describe the morphology of the amastigote and promastigote form of
Leishmania donovani
2. Discuss the laboratory diagnosis of a case of leishmaniasis.
ANSWERS TO INTEXT QUESTIONS
42.1
1. Reticuloendothelial cells
2. Sand fly
3. Vacuole
4. Sand fly
5. Leishmania Donovani
42.2
1. (c)
2. (a)
3. (b)
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