CHAPTER 5

Sporozoa
Sporozoans producing spore-like cyst
Toxoplasma gondii, Cryptosporidium, Isospora belli, Cyclospora cayetanensis & Sarcocystitis
species.Have asexual (schizogony) & sexual (gametogony) reproduction.
Toxoplasma gondii
Causes zoonotic disease. Parasite of intestinal tract in cat family.
Epidemiology:
Distribution: Worldwide. Most cases in France & Scandinavian countries (N.Europe-Denmark,
Finland, Sweden etc). 30-40% of world population seropositive.
Hosts:
Definitive host: Domestic & wild cats. Oocysts formed.
Paratenic hosts: Not needed for the development of the parasite, but serves to maintain the life
cycle. (Humans, rodents, pigs, cattle)
Transport hosts: Cockroaches, earthworm, flies.
Reservoirs: Both definitive & Paratenic hosts.
Transmission:
Consumption of improperly cooked meat (pork, Beef), contaminated vegetables & water with cat
faeces containing oocysts.
Transplacental: Occurs in 1 per 2000 live births. Meat handlers, hunters, pregnant women,
children at high risk.
Morphological forms:
Trophozoite (Tachyzoite/Endozoite) & Tissue cyst: Asexual forms. (Schizogony). Found in
intermediate hosts (Humans , other mammals) & definitive hosts (Cats)
Oocyst.: Formed by sexual reproduction (Gametogony & sporogony). Seen in definitive hosts
(Cats) only.
Infective stages:
Trophozoites(Tachyzoites),Bradyzoites (Found inside tissue cysts),Sporozoites (Found in
oocysts)
Trophozoite: (Tachyzoite/Endozoite)
Cresentric or oval shaped. Found intracellularly in all tissues & organs (except non-nucleated
RBCs).One end rounded & other tapering. 4-8µm X 2-3µm.Ovoid nucleus near round
end.Rapidly proliferating trophozoite in acute infection is called tachyzoite.In Giemsa stain,
cytoplasm is azure blue & nucleus red.
Tissue cyst:
10-100µm. Contains over 3,000 spore-like bradyzoites or cystozoites.Bradyzoites similar to
trophozoite morphology except contains lot of amylopectin & smaller.Formed in chronic phase
& found in muscles, brain & other tissues.Bradyzoites initiate new infection in mammals
ingesting cyst-bearing tissue.
Oocyst:
Develop only in definitive host. Excreted in stools.Fresh oocyst non-infectious. Become infective
in soil or water after1-5days. (Sporogony) ,10-12µm, spherical or ovoid. Contains sporoblast &
resistant cyst wall.Sporoblast divide into 2 sporocysts within each oocyst. Each sporozocyst give
rise to 4 sporozoites. The mature oocyst with 8 sporozoites is infective to cat, birds, rodents,
humans etc.
Life cycle:
1. Enteric cycle: (In definitive host-cat)
2. Exoenteric cycle: (In mammals-rodents, man)
Enteric cycle: (Development in cat)
Ingestion of mature oocyst (containing sporozoites) or flesh (Rodents/mammals) containing
tissue cysts with bradyzoites.
.

Cyst wall disrupted by peptic/tryptic digestion, releasing sporozoites from oocysts & bradyzoites
from tissue cyst, which penetrate small intestinal epithelial cells.

Both sporozoites & bradyzoites develop to form tachyzoites (Asexual). Tachyzoites multiply
locally.

Some tachyzoites mature

to form male & female Some tachyzoites migrate to

extraintestinal sites (Muscles, brain etc) except RBCs,

via blood & lymphatics

gametocytes (sexual) Some tachyzoites are pahgocytosed. Surviving

tachyzoites becomes encysted & form tissue

cysts.

.Male & female gametocytes fertilize

in intestine to form oocyst & excreted in stool.

Development in man:

Follows ingestion of contaminated or undercooked food. Infection also occurs by Congenital

transmission, blood transfusion or organ transplant.Life cycle similar to cats except there is no
sexual stage (no formation of gametocytes or oocyst). Tissue cysts formed in brain, retina,
skeletal muscles, lungs, heart etc.
Exoenteric cycle

Clinical disease:
A. In Immunocompetent:
IP: 1-3wks.
Usually causes mild or asymptomatic infection.
Acute infection resembles infectious mononucleosis. Lymphadenopathy is common. Rarely
meningitis, myocarditis, encephalitis, pneumonia are seen.
Congenital infection in 1st trimester leads to still birth, microcephaly, hydrocephaly. Late
infections can cause retinitis & mental defects.
Occular toxoplasmosis mostly congenital origin. Latent infections may reactivate in adolescents
causing retinochondritis & impairment of vision.
A. In Immunodeficient:
Reactivation in HIV leads to spastic paralysis, blindness, myocarditis, meningitis or
encephalitis.
Lab diagnosis:
Demonstration of tachyzoites & cysts, antigen detection, Serology.
Microscopy:
Geimsa stain of CSF, body fluid will show crescent shaped, round on one side other side pointed,
4-8µm long trophozoites (tachyzoites) with azure blue cytoplasm & red nucleus. Section from
tissue cysts also shows numerous bradyzoites with similar shape but smaller.
Culture:
Can be grown intraperitoneally in white mice by inoculating infected tissue or fluids.
Antigen detection: By PCR. From CSF, Blood & tissues.
Serology:
Detection of IgM & IgG by ELISA, IIFA or latex agglutination tests.
Positive IgM indicates primary infection. Rising IgG antibody titer indicates active infection.
In pregnancy, positive IgM, rising IgG titers indicates ongoing infection. Pregnancy to be
terminated. A high and stable IgG titer indicates infection acquired recently. < 1% risk to foetus.
In congenital infection positive IgM & rising IgG indicative.
Sabin-Feldman dye test:
First test for toxoplasmosis developed in 1948. Not in use now.T.gondii trophozoites cultivated
in mice peritoneum is used. Equal volumes of patient’s serum & suspension of trophozoites plus
a normal human serum are mixed Several tubes with different dilutions of patient’s serum done.
The tubes are incubated at 370c, for 1 hr. One drop of alcoholic solution of methylene blue
(pH11.0) is added to each tube & reincubated.One drop from each tube is examined under
microscope.
Results:
If patient’s serum contains abs. trophozoites are inactivated & killed by complement mediated
lysis from normal serum. Trophozoites which are not destroyed are stained by methylene blue.
The dilution at which 50% of trophozoites are stained is the titer. Significant titer is 4. Test is
positive by 1-2 wks after infection.False positive results seen in sarcocystis, T.vaginalis,
Trypanosoma lewsii infections.
Treatment: Pyrimethamine, sulphadiazine, spiramycin.

Cryptosporidium parvum
Causes diarrhoeal disease. Common in immunocompromised.
Species: C.parvum (Host: House mouse), C.felis (cat) etc.
Epidemiology:
Distribution: Cosmopolitan & commonly found in domestic animals & man.
Transmission: Faeco-oral route. By ingestion of oocysts in contaminated water. Resistant
chlorination. Zoonotic disease. Verternarians, animal handlers at risk.
Life cycle:
Oocysts infective stage. Oocyst contains 4 sporozoites which is released & penetrates intestinal
epithelium.

Sporozoites form trophozoites & schizonts. Schizonts rupture releasing merozoites.

Some merozoites form micro &macro gametocyte, fertilize to form oocyst.

Thick walled oocyst excreted in faeces (80%). Thin walled oocyst (20%) excysts internally &
cause autoinfection.
Clinical disease:
Asymtomatic or self-limiting watery diarrhoea. In immunocompromised (AIDS) causes severe to
fatal disease.
Lab diagnosis:
Specimen: Stool, biopsy (from intestine, gall bladder).
Microscopy:
Wet mount: (Direct or formal ether concentration): Oocyst or sausage shaped sporozoites may be
observed.
Staining: Acid fast in nature. Modified Ziehl-Neelsen’s stain (0.5% H 2SO4) is done. Oocysts
appear Small, oval, pink-red stained, 5µm in diameter.
Indirect immunofluorescent staining can be done.
Newer tests: PCR from stool.
Serology: Abs. detection by ELISA.
Treatment:
Spiramycin, clindamycin.

Isospora
Species: Isospora belli, I.natalensis, I.hominis.
Epidemiology:
Worldwide distribution: More in central & S.America, Africa & S.E.Asia.
Transmitted faeco-orally. More incidence in HIV.
Life cycle:
Similar to Cryptosporidium. Mature oocyst contains 8 sporozoites. No auto-infection
Clinical disease:
Usually asymtomatic. Mild self limiting diarrhoea. [ I.hominis may can lesions in muscles].
Extraintestinal spread common in AIDS.
Lab diagnosis:
Stool examination by wet mount & Modified Ziehl-Neelsen Staining. Acid fast in nature.
Duodenal aspirate, intestinal biopsy & muscle biopsy also done.
Oocyst is elongated, ovoid, measures 25-23µm X 12-15µm. Contains 2 spherical sporoblasts.
Cyclospora cayetanensis:
Worldwide distribution. Seen in reptiles, birds & animals. Infection occurs by consumption of
contaminated water with sporulated oocysts. Causes watery diarrhoea, fever, abdominal pain.
Responsible for Traveller’s diarrhoea. In immunocompetant self-limiting, relapses common.
Oocysts are 8-10µm. Oocysts mature in environment after excreted from faeces. They are acid
fast.
Sarcocystis species:
Causes mild form of diarrhoea. Severe for in AIDS.

Microspora
Microspora are acid fast, forms spore and mainly causes opportunistic infections in AIDS.
Medically important genera are Enterocytozoon, Encephalitozoon etc.
Spore is the infective stage. Infection occurs by consumption of contaminated food & water.
Spores are spherical & 6µm. Most of them cause diarrhoea in AIDS. Some also cause
keratoconjunctivitis, sinusitis, bronchitis, pneumonia etc.Organism can be detected by modified
ZN stain from stool, urine , CSF. ELISA & PCR also used.