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PARASITOLOGY LABORATORY 11 – Trypanosomes etc.

USTMED ’07 Sec C – AsM, AnnePonce; Photos provided by JV.N.

Gomori methenamine-
Female mosquito head silver nitrate stain.

Cysts in human lung


impression smear

Male mosquito head

PNEUMOCYSTIS CARINII Pneumocystis carinii

H&E stained section of human


CLASSIFICATION
– Protozoa
Alveoli appear to be filled with a
DISEASE
pink-staining, foamy material,
– Pneumocystosis, interstitial plasma cell pneumonia
Neither cysts nor organisms can be
GEOGRAPHIC DISTRIBUTION seen in this stain
– cosmopolitan

LOCATION IN HOST
– lung tissue

MORPHOLOGY
• Trophozoites: small, 1-5µ in diameter, with ovoid or
amoeboid appearance
• Cysts: 3.5-7.0µ in diameter, have an outer membrane of
variable thickness and usually contain 4 to 8 nuclei,
each of which measures 1-2 µ in diameter

LIFE CYCLE
- Although the life cycle has not been clearly elucidated,
Pneumocystis probably exists in the environment and in TRYPANOSOMA SPECIES
the lungs of rodents and humans without causing
apparent disease. However, in hosts that are
immunosuppressed for extended periods of time or in 145 & 146 Trypanosoma cruzi in human blood film. The causative
premature and malnourished infants, the organisms may agent is T. cruzi. It occurs characteristically in blood films as short
proliferate in lung tissue, leading in consolidation, “C” or “S” shaped trypomastigotes with a prominent kinetoplast.
dyspnea, and even death. It is otherwise monomorphic (x900)
DIAGNOSIS
- Usually established by demonstrating organisms in lung
tissue. Specimens obtained by lung biopsy, lung
aspirates, or bronchial brushings are more satisfactory
for examination than are sputum or bronchial washings
because more organisms are likely to be detected.
Although a variety of stains (Giemsa’s, methylene blue,
hematoxylin-eosin) have been used, the Grocott
modification of the Gomori methenamine-silver nitrate
stain is especially useful because organisms stain darkly
147 Typical vector biting.
and are readily identifiable against an unstained
Reduviid bugs (also known as
background.
“assassin” or “kissing” bugs),
particularly in the genera
- DIAGNOSTIC PROBLEMS. Obtaining a suitable specimen Triatoma, Rhodnius and
for examination is essential. Pneumocystis must be Panstrongylus, transmit T.
differentiated from yeasts, fungi and other organisms cruzi while feeding, not by
inoculation but by fecal
contamination.
Pneumocytsis carinni cyst Involved in:
1. Upper and Lower Respiratory Tract Infections 148 & 149 Life cycle in vector. Trypomastigotes picked up in the
2. Eye Infections blood meal transform to epimastigotes in the midgut of the bug.
They reproduce as epimastigotes, then pass to the hindgut where
they transform again to the infective stages (metacyclic
trypomastigotes). The figures show epimastigote stages in the
vector.
Characteristic cysts stained with
Gomori methenamine-silver
nitrate are seen at high
magnification
Trypanosoma gambiense
Figure 1. This is a thin blood film
containing the trypomastigote stage of TOXOPLASMA GONDII
T. gambiense. These trypomastigotes,
indistinguishable from those of T.
rhodesiense, have a small kinetoplast Toxoplasma gondii
near the somewhat blunted, posterior bradyzoite
end, and a conspicuous, undulating
membrane with a flagellum. At the
extreme right margin note the two
dividing forms; this is characteristic Cyst in H & E stained tissue section of
for the African trypanosomes, but is mouse brain
not seen in T. cruzi.

Trypanosoma cruzi trypomastigote cyst

Figure 2. “Broad: form of T. cruzi Toxoplasma gondii


trypomastigote in mouse thin blood Giemsa-stained trophozoites in tissue
film. Note the large kinetoplast culture
close to the short, pointed,
posterior tip of this organism.
Broad forms, such as this usually
assume an S or C shape in stained
preparations. The centrally located
nucleus is subspherical, and the
undulating membrane inconspicuous. T. cruzi does not undergo
division in the trypomastigote stage.

Trypanosoma cruzi Toxoplasma gondii

Figure 3. “Slender” form of T. cruzi Giemsa-stained trophozoites in


trypomastigote in mouse thin blood film. peritoneal fluid of mouse called
Slender forms are commonly found during Tachyzoites found in tissues and
acute infections. Note the elongate nucleus fluids during the acute stage of
and the large kinetoplast followed by a infection.
long, pointed, posterior tip. The undulating
membrane may be inconspicuous in stained
preparations.
Toxoplasma gondii
Giemsa stained cyst in an impression smear

Trypanosoma cruzi

Figure 4. Two typical C forms are visible


in this mouse thin blood film of
trypomastigotes of T. cruzi. Note the
very large kinetoplasts that seem to
bulge from the body at the posterior
end. In contrast, the kinetoplasts in the
African trypanosomes are small and CRYPTOSPORIDIUM
subterminal
Cryptosporidium oocyst

Trypanosoma cruzi
Figure 5. In this illustration of T.
cruzi epimastigotes in culture,
note the small, S-shaped
trypomastigote above and to the
left of center. It has a large
kinetoplast near the posterior
end and a flagellum that extends 15-26, 27 Cryptosporidium parvum. Oocysts, Feces, Acid-fast and
along its length, projecting fluorescence stain. In acid-fast stained preparations, the oocyts of
beyond the anterior end. An C. parvum appear round to oval, measuring approximately 4 to 6
undulating membrane is present um in diameter. Several black granules can be observed, usually in
between the flagellum and the body of the organism. This form – the periphery of the cyst. Under fluorescence, the wall of the
the infective stage for humans – is found in cultures and in feces cysts may appear smooth or wrinkled.
of infected triatomid bugs.
Cryptosporidium oocyst
Trypanosoma cvnasi

ISOSPORA BELLI

Trypanosoma sp. Isospora belli, immature oocyst


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