2. Toxoplasma gondiiToxoplasma gondii Worldwide Zoonotic
parasite; Toxoplasma is an opportunistic pathogen. Infects animals,
cattle, birds, rodents, pigs, and sheep. and humans. Causes the
disease Toxoplasmosis. Toxoplasmosis is leading cause of abortion
in sheep and goats. Intracellular parasite. Final host (Felidae
family, cat) Intermediate host (mammals ) Toxoplasmosis 1. All
parasite stages are infectious. 2. Risking group: Pregnant women,
meat handlers (food preparation) or anyone who eats the raw
meat
3. Definitive (final) host. Domestic cats, who pick up the
organism from eating infected rodents. Asexual and sexual division
is intracellular. Oocysts in feces. Intermediate host.. Asexual
tissue cycle. Motile, disease producing phase = tachyzoites.
Non-motile slow phase in tissue cyst = bradyzoites. Humans
(Mammals) Cats (Mainly domestic and wild cats)
4. Toxoplasma gondiiToxoplasma gondii exists in three
formsexists in three forms All parasite stages are infectiousAll
parasite stages are infectious.. 1. Tachyzoites 2. Tissue cysts
(bradyzoit) 3- Oocysts
5. Tachyzoite stageTachyzoite stage Rapidly growing stage
observed in the early stage of infection. (Acute phase) habits in
the body fluid. Crescent-shaped. One end is more pointed than the
other subterminal placed nucleus. Asexual form. Multiplies by
endodyogeny. It can infect phagocytic and non-phagocytic,
cells.
6. BradyzoitesBradyzoites Are slow-growing stage inside the
tissue cysts. Bradyzoites mark the chronic phase of infection.
Bradyzoites are resistant to low pH and digestive enzymes during
stomach passage. Protective cyst wall is finally dissolved and
bradyzoites infect tissue and transform into tachyzoites.
Bradyzoites are released in the intestine and are highly infective
if ingested.
7. Oocysts in the feces of catOocysts in the feces of cat Cat
ingests tissue cysts containing bradyzoites. Gametocytes develop in
the small intestine. Sexual cycle produces the oocyst which is
excreted in the feces. Oocysts appear in the cats feces 3-5 days
after infection by cysts. Oocysts require oxygen and they sporulate
in 1- 5 days.
8. The Oocyst The oocyst is noninfectious before sporulation.
Unsporulated oocysts are subspherical to spherical. Sporulated
oocysts are subspherical to ellipsoidal. Each oocyst has two
ellipsoidal sporocysts. Each Sporocyst contains four sporozoites .
Shedding occurs 3-5 days after ingestion of tissue cysts Sporulated
oocyst remain infective for months .
9. Life cycle of T. gondii
10. Sources of infection : Contaminated water or food by
oocysts Ingestion of tachyzoites and bradyzoites (cysts) in flesh
of infected host. Undercooked meat. Mother to fetus. Organ
transplant (rare). Blood transfusion (rare). Toxoplasma
Transmission
13. Prophylaxis 1- Individuals at risk, particularly pregnant
women, children, and immunocompromised persons should avoid contact
with cat and its feces. 2- Proper cooking of meal. 3- Proper
washing of hands and washing of vegetables and fruits before
eating. 4-Blood or blood products from seropositive persons 5-
should not be given and screening for T. gondii antibody 6- should
be done in all blood banks.
14. control It is difficult to control toxoplasmosis because of
wide range of animal reservoirs. Currently, there is no effective
vaccnine available for humans. A genetically engineered vaccine is
under development for use in cats. Treatment Congenital infection
is treated with pyrimethamine and sulfadiazine. For primary
prophylaxis Trimethoprimsulfamethoxazole is the drug of choice
16. Morphology Oocysts of I. belli are elongatedovoid and
measure 25 m 15 m. Each oocyst is surrounded by a thin smooth 2
layered cyst wall Immature oocyst seen in the feces of patients
contain two sporoblasts. The oocysts mature outside the body.
17. On maturation, the sporoblast convert into sporocysts. Each
sporocyst contain 4 crescentshaped sporozoites The sporulated
oocyst containing 8 sporozoites is the infective stage of the
parasite. A B Oocysts of Isospora belli. A. Immature cyst; B.
Mature cyst
18. Life Cycle I. belli completes its life cycle in one host.
Man gets infection by ingestion of food and water contaminated with
sporulated oocyst. When a sporulated oocyst is swallowed, 8
sporozoites are released from the 2 sporocysts in the small
intestine and invade the intestinal epithelial cells. In the
epithelium, the sporozoites transform into trophozoites, which
multiply asexually (schizogony) to produce a number of
(merozoites). The merozoites invade adjacent epilhelial cells to
repeat asexual cycle.
19. Some of the trophozoites undergo sexual cycle (gameto gony)
in the cytoplasm of enterocytes and transform into macrogametocytes
and microgametocytes. After fertilization, a zygote is formed,
which secretes a cyst wall and develops into an immature oocyst.
These immature oocysts are excreted with feces and mature in the
soil. Incubation period: 14 days.
20. Life Cycle of Isospora belli
21. Clinical Features Infection is usually asymptomatic.
Clinical illness includes abdominal discomfort, mild fever,
diarrohea, and malabsorption. The diarrohea is usually watery and
does not contain blood or pus and is selflimiting. However,
protracted diarrohea, lasting for several years can be seen in
immunocompromised persons, particularly in the human immunodefi
ciency virus (HIV) infected.
22. Indirect evidence High fecal fat content. Presence of fatty
acid crystals in stool. Presence of CharcotLeyden crystals in
stool. Direct evidence It may be diffi cult to demonstrate the
transparent oocyst in saline preparation of stool. Stool
concentration techniques may be required when direct wet mount of
stools are negative. The staining technique used are Modified
ZiehlNeelsen stain or Kinyoun acid fast staining of stool smear. In
these methods, pink colored acid fast large oocyst (>25 m) can
be demonstrated. The stool smear can also be stained by auramine
rhodamine and Giemsa stains. Laboratory Diagnosis 1-Stool
Examination
23. 2- Duodenal Aspirates After repeatedly negative stood
examinations, duodenal aspirate examination or enterotest can be
performed to demonstrate oocyst. 3- Intestinal Biopsy Upper
gastrointestinal endoscopy may provide biopsy specimens for
demonstration of oocysts. 4-Others Eosinophilia, which is generally
not seen with other enteric protozoan infections, is detectable in
case of isosporoloasis
24. Treatment No treatment is indicated in self limiting
infection in immunocompetent persons. Immunodefecient patients with
diarrhea and excreting oocysts in the feces should be treated with
cotrimoxazole (trimethoprimsulfamethoxazole) in a dose of 2 tablet,
4 times a day for 10 days followed by 2 tablets 2 times a day for 3
weeks. For patients intolerant to sulfonamides, pyrimethamine 5075
mg/day is given.Relapses can occur in persons with AIDS and
necessitate maitainance therapy with cotrimoxazole 1 tablet thrice
a week.
26. Morphology The infective form of the parasite is oocyst.
The oocyst is spherical or oval and measures about 5 m in diameter.
Oocysts does not stain with iodine and is acid fast. The wall of
the oocysts is thick, but in 20% cases, wall may be thin. These
thin walled oocysts are responsible for autoinfection. Both thin
walled and thick walled oocyst contain 4 crescent shaped. Oocyst
can remain viable in the environment for long periods, as it is
very hard and resistant to most disinfectants and temperature upto
60C. It can survive chlrorinated water, but sequential application
of ozone and chlorine has been found effective in eliminating the
cysts.
27. The parasite complete its life cycle, sexual and asexual
phases in a single host (monoxenous) Suitable host: Man.
Reservoirs: Man, cattle, cat, and dog. Mode of transmission: Man
acquires infection by: Ingestion of food and water contaminated
with feces containing oocysts Autoinfection. Infective form:
Sporulated oocysts. The oocyst contains 4 sporozoites, which are
released in the intestine.
28. Life Cycle
29. Pathogenicity and Clinical Features Clinical manifestations
of c. parvum infection vary depending upon the immune status of the
host. Infection in healthy immunocompetent persons may be
asymptomatic or cause a selflimiting febrile illness, with watery
diarrhea in conjunction with abodminal pain, nausea, and weight
loss. It can also cause childhood and travellers diarrhea, as well
as waterborne outbreaks.
30. In immunocompromised hosts, especially those with AIDS and
CD4+ T cell counts below 100/uL, diarrhea can be chronic,
persistent, and remarkably profuse, causing significant fl uid and
electrolyte depletion, weight loss, emaciation, and abdominal pain.
Stool volume may range from 1 to 25 L/day. Billary tract
involvement can manifest as right upper quadrant pain, sclerosing
cholangitis, or cholecystitis.
31. Laboratory Diagnosis Stool Examination Diagnosis is made by
demonstration of the oocysts in feces. A direct wet mount reveals
colorless, spherical oocyst of 45 m, containing large and small
granule. The oocysts are diffi cult to visualize in unstained wet
preparations. Modifi ed acid fast staining is the method of choice
and by this method oocysts appear as red acidfast spheres, gainst a
blue background . Yeast closely resembles oocysts of c. parvum in
shape and size but can be differentiated by using acidfast stain,
as they are not acidfast and appear blue in color. The staining can
also be used for demonstration of oocysts in other specimens like
sputum, bronchial washing, etc.
32. If oocysts load is less and cannot be demonstrated even
after examination of 3 wet mounts of stool specimen, concentration
techniques like Sheather's sugar floatation technique and zinc
sulfate floatation technique can be applied. Fluroscent staining
with auraminephenol or acridine orange has also been reported to be
a useful technique. Definitive identification can be made by
indirect immunofl uroscence microscopy using specific
antibody.
33. Histopathological Examination: Cryptosporidia can also be
identified by light and electron microscopy at the apical surface
of intestinal epithelium from biopsy specimen of the small bowel
(jejunum being the preferred site). Serodiagnosis: Antibody
persists for at least an year and can be demonstrated by ELISA or
immunofl uroscence. An ELISA for detection of cryptosporidium
antigens in stools using monoclonal antibody has also been
developed and is highly sensitive and specifi c. Molecular
Diagnosis For seroepidemiological study, western blot technique is
employed by using a 17KDA and 27KDA sporozoite antigen. PCR
technique has also been applied to detect viable cysts
34. Supportive therapy with electrolytes and fluids and early
antiretroviral therapy in AIDS patients.