BLOOD PROTOZOA

II BPT

Dr Ekta Chourasia
Microbiology
 Protozoa - Blood
protozoa
Plasmodiu Malaria
m

Babesi Babesiosi
a s

Leishmani Leishmaniasis (Visceral,

a Cutaneous)

Trypanaso African sleeping sickness, Chagas

ma disease

Toxoplasma Toxoplasmosis (congenital

gondii infections)
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 Genus Plasmodium
Consists of 4 species:
 P. vivax
 P. falciparum
 P. malariae
 P. ovale

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 Transmission & Life Cycle
Definitive host Female Anopheles mosquito

Intermediate host Man

Infective form Sporozoites

Portal of entry Skin

Mode of transmission Bite of an infected mosquito

Site of localization First in liver cells & then in

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RBCs Dr Ekta, Microbiology, GMCA
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 Morphological forms seen in Humans
In liver:
• Sporozoites
• Pre erythrocytic schizonts
•
Merozoites – infect RBCs

In RBCs :
1. Trophozoites – ring form
2. Schizonts
3. Merozoites – released by the rupture of schizonts
– infect other RBCs
4. Gametocytes – micro and macro gametocytes

12/03/08 Dr Ekta, Microbiology, GMCA

 Morphological forms seen in Mosquito
Further differentiation & development of
gametocytes take place in mosquito
• Macro gametes (female gametes) – each
macro gametocyte develops in to one macro
gamete in the mid gut of mosquito
• Micro gametes (male gametes) – one micro
gametocyte produces 6 to 8 micro gametes
by exflagellation.
• Zygote – Ookinete – Oocyst – rupture –
release of Sporozoites – predilection to
salivary glands.

12/03/08 Dr Ekta, Microbiology, GMCA

 Incubation period
P. vivax
P. ovale 10 to 14 days
P. falciparum

P. malariae 18 days to 6
weeks

12/03/08 Dr Ekta, Microbiology, GMCA

 Pathogenicity
Infection causes intermittent fever – Malaria
Each of the 4 species causes a characteristic
fever:
P. vivax Benign tertian/ vivax malaria
P. falciparum Malignant tertian/ falciparum
malaria, black water fever
P. malariae Quartan malaria
P. ovale Ovale malaria

12/03/08 Dr Ekta, Microbiology, GMCA

 Clinical Features
Series of febrile paroxysms – fever is caused
by the release of merozoites & toxins from
ruptured erythrocytic schizont which in turn
causes the release of cytokines.
Quartan malaria – every 72 hrs
Tertian malaria - every 48 hrs

* each paroxysm has 3 stages - cold stage

(rigors), hot stage (high temp., body & joint
pains, vomiting & diarrhoea) and perspiration
stage (fall in temp.)

12/03/08 Dr Ekta, Microbiology, GMCA

 Clinical Features
Anaemia – due to breakdown of RBCs,
particularly occurs in falciparum malaria

Splenomegaly – all forms

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 Falciparum malaria
Severe falciparum malaria is associated
with
1. Pernicious malaria /cerebral malaria
2. Blackwater fever
3. Anaemia
4. Hypoglycaemia
5. Complications in pregnancy

12/03/08 Dr Ekta, Microbiology, GMCA

 Malaria caused by P.vivax,
P.ovale & P.malariae
 Rarely life threatening
 Relapses/ recurrences are a feature

Recurrences in Malaria
 May result from – reinfection or
- due to persistence of infection
- Occurs due to a special form of
parasites called hypnozoites.

12/03/08 Dr Ekta, Microbiology, GMCA

 Laboratory diagnosis of Malaria

Specimen: peripheral blood smears

thick and thin blood smears

Leishman or Giemsa stain

Trophozoites (ring forms) or gametocytes are

seen within RBCs

Quantitative Buffy Coat (QBC) examination

P falciparum antigen detection (ELISA)

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 Making of Thin & Thick films

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 Rapid Diagnostic tests
HRP2 tests

detection of P.falciparum

Two types of test – ParaSight F
- ICT Malaria Pf

pLDH test e.g. OptiMAL test

 Detection of P.falciparum & P.vivax
 Produced by all human malarial parasites
 Differentiation of species is based on antigenic
differences between pLDH isoforms.

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 ParaSightF test

Optimal test

ICT Malaria Pf / Pv

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 Toxoplasma gondii
Definitive host Cat

Intermediate host Man, sheep, birds

Infective form Matured oocyst / tissue cyst /

tachyzoites

Mode of Ingestion / intrauterine / blood

transmission transfusion/ improper handling of cat
litter

Site of localization Any organs (RE system)

Eye & Brain

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12/03/08 Dr Ekta, Microbiology, GMCA
 Laboratory Diagnosis –T gondii

Specimens Impression smear of LN, bone marrow, spleen, brain

biopsy, blood, sputum, CSF
Microscopy Giemsa stain, tachyzoites or cysts

Serology ELISA / IFA – 16 fold rise in Ab titre: Acute infection

Sabin-Feldman dye test: inhibition by antibody of the
staining of tachyzoites by alkaline methylene blue

Prenatal diagnosis Fetal blood for IgM Ab / PCR

12/03/08 Dr Ekta, Microbiology, GMCA

 Haemoflagellates

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 Trypanosomes

T. brucei subspecies gambiense West African sleeping

(Chronic) sickness

T. brucei subspecies rhodesiense East African sleeping

(Acute) sickness

T. cruzi (acute and chronic) American trypanosomiasis

Chagas disease

12/03/08 Dr Ekta, Microbiology, GMCA

 T. brucei

Definitive host Man

Intermediate host Tsetse fly

Infective form Metacyclic Trypomastigote

Mode of transmission Bite of infected tsetse fly

Site of localization CNS

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 Clinical features Sleeping Sickness

Trypanosoma chancre at the site of bite

Winter bottom’s sign: prominent cervical

lymphadenopathy

Meningoencephalitis - Apathetic, confused,

comatose

12/03/08 Dr Ekta, Microbiology, GMCA

 T. cruzi
Definitive host Man

Intermediate host Reduviid bug

Infective form Metacyclic Trypomastigote

Mode of transmission Feces of bug rubbed into site of

bite / contamination of conjunctiva
or other exposed mucous
membranes with fingers

Site of localization Autonomous nervous system of

heart / GIT
Infect cardiac, smooth and skeletal muscle, reticuloendothelial cells and neuroglial cells
12/03/08 Dr Ekta, Microbiology, GMCA
 Clinical features

Chagoma chancre at the site of bite

Acute Chagas disease

Romana’s sign: unilateral edema of face with
conjunctivitis and swelling of upper & lower eyelids
 Fever, splenomegaly, anasarca, meningoencephalitis
Chronic Chagas disease
 Cardiomyopathy, AV block, CCF

Megaesophagus / Megacolon

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Laboratory Diagnosis – Trypanosomiasis
Specimens Blood, CSF, Aspirates (LN)

Microscopy Trypomastigotes in blood

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 Leishmania

Definitive host Man

Intermediate host Sand fly (Phlebotomus)

Infective form Promastigote
Mode of transmission Bite of infected sand fly

Site of localization Reticuloendothelial system

12/03/08 Dr Ekta, Microbiology, GMCA

 Leishmania
Three major species:
 L. donovani – kala azar/ visceral leishmaniasis

 L. major – cutaneous leishmaniasis

 L. braziliensis – mucocutaneous leishmaniasis

12/03/08 Dr Ekta, Microbiology, GMCA

 Clinical Types
Visceral leishmaniasis - fever, malaise, weight loss,
anaemia and swelling of the spleen, liver, and lymph
nodes

Cutaneous leishmaniasis - causes 1-200 simple skin

lesions which self-heal within a few months but which
leave unsightly scars

Mucocutaneous leishmaniasis - infection begins with skin

ulcers which spread, causing dreadful and massive
tissue destruction, especially of the nose and mouth

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 Laboratory Diagnosis
 Demonstration of parasite in clinical specimen:
 Microscopy
 Culture
 Animal inoculation

Demonstartion of antibodies using

 Specific leishmanial Ag – ELISA / IFA / Agglutination
 Non-specific Ag – CFT

 Non-specific serum tests

 Aldehyde test (Napier’s)
 Chopra’s Antimony test

Absence of hypersensitivity to leishmanial Ag

Contributory lab findings – anemia, leucopenia, neutropenia

12/03/08 Dr Ekta, Microbiology, GMCA

 1. Demonstration of parasite in Clinical
Specimen
Clinical specimens:
 Peripheral blood

Bone marrow aspirate

Spleen aspirate

Lymph node aspirate

Microscopy: Leishman, Giemsa or Wright’s

stain - Amastigotes within macropahges

Culture: NNN (Novy, MacNeal, Nicolle) medium for 7

days – promastigote form

Animal inoculation: Hamster - Animal kept at 23-26°C

12/03/08 Dr Ekta, Microbiology, GMCA

 2. Absence of Hypersensitivity to
Leishmanial Ag

Montenigro test – 0.1 ml of killed promastigote

Ag I.D. Result read after 72 hrs.

Positive in Dermal leishmaniasis & recovered cases
of Kala Azar.

 Negative in active cases of Kala Azar.

12/03/08 Dr Ekta, Microbiology, GMCA