Parasitology (dra Madrid)

Coccidians
20 February 08
November 07
Coccidians Isospora belli (Life cycle)

▪At time of excretion, immature oocyst contains usually 1

sporoblast
Coccidian Parasites
▪Sporoblast divides into two, secrete a cyst wall sporocysts
▪Members of the class Sporozores (Phylum Apicomplexa) ▪Sporocyst divide to produce sporozoites
▪Obligate intracellular protozoans ▪Infection occurs in ingestion of sporocyst containing
oocyst(oocyst containing 2 sporocyst with each containing 4
▪Gliding motility sporozoites)
▪Reproduction ▪Sporocyst excyst in small intestinerelease sporozoites
o Sexual phase: sporogony, gametogony enter epithelial cells of distal duodenum and proximal ileum
o Asexual phase: schizogony, merogony o Invade new epithelial, continue asexual
(trophozoites, schizonts, merozoites) multiplication
▪Species included: ▪Trophozoites develop into schizonts which contain multiple
o Isospora merozoites
o Sarcocystis o Rupture of schizonts  release of merozoites 
o Cryptosporidium invade new epithelial cells and continue of
o Cyclospora asexual multiplication
o Toxoplasma
o Pneumocystis
▪After 1 wk, sexual stage begins – male and female
gametocytes
o Microgametocytes fertilize macrogametocytes

Isospora belli ▪Fertilization product: oocyst excreted in stool

▪Coccidian parasite
▪Infects epitheial cells of the small intestine
▪Infections usually in immunocompromised patients
o Oocyst mature in 48 hrs  oocyst initially has 1
sporoblast  divides into 2 sporoblast 
develop into sporocysts (2)
o 4 long and slender sporozoites develop within
each sporocyst

▪Geographic distribution ▪Oocyst of isospora contains 2 sporocysts, each with 4

o Worldwide, in tropical and subtropical areas
o High prevalence: Haiti and other tropical areas
with poor sanitary conditions
▪Transmission: food and water
▪Dse common in children and male homosexuals especially
those with AIDS
▪Definitive hosts: human and animals
Pathogenesis
▪Parasites develop and multiply in the epithelial cells of the
small intestine
▪Trophozoites develop into schizonts(contain merozoites);
rupture of schizonts release the merozoites  invade new
epithelial cells, and continue the cycle of asexual
multiplication
▪After 1 week sexual stage begins: devt of male and female
gametocytes fertilization  devt of oocysts excreted in
the stool
▪Immature oocyst contains usually 1 sporoblast  further
maturation  sporolast divides in 2 (oocyst now contains 2
sporoblasts); sporoblasts secrete a cyst wall and becomes
sporocysts  sporocysts divide 2x to produce 4 sporozoites
each
▪Infection occurs by ingestion of sporocyst-containing
oocysts; sporocysts excyst in small intestine, release their
sporozoites  invade epithelial cells and initiate schozogony
sporozoites
Isospora belli: Clinical Manifestations
▪Infections often asymptomatic even with excretion of
oocysts
▪Symptomatic infections:
o Diarrhea: intermittent for months
o Fever, malaise, anorexia, abdominal pain,
flatulence
o Malabsorption along with weight loss and rarely
death
o In immunocompromised hosts, severe diarrhea
o Eosinophilia
▪Intestinal biopsies show:
o Mucosal lesions of shortened villi
o Hypertrophied crypts
o Infiltration of lamina propria with pmn
leucocytes especially eosinophils
▪Dissemination to other organs: seen in AIDS patients
▪Prognosis is good but infection may last for months
Isospora belli: diagnosis
▪Detection of oocysts in feces
▪Oocysts in stools: ellipsoidal shape

YNA, joy, cams, shar – 2B1 1 of 7

 Parasitology – Coccidians by Dra Madrid
Isospora belli: treatment
▪Asymptomatic infections: bed rest and a bland diet
▪Trimethoprim-sulfamethoxazole: drug of choice
Sarcocystis
▪Worldwide: infects humans and animals
▪Disease: sarcosporidiosis and sarcocystosis
▪Complex life cycle – involves intermediate hosts
▪Definitive hosts: man and carnivores (dog)
▪Human definitive hosts for 2 species:
o Sarcocystis hominis
o Sarcocystis suihominis
▪Humans can also act as intermediate hosts
▪Take several forms
▪Simplest is called the zoite
o Banana shaped cell with a pointed end
equipped for entering host cells
▪Sporocysts are composed of 4 zoites
o Oval; can survive on ground and infect
intermediate hosts
▪Sporozoites are formed from sporocysts
▪Sarcocysts are formed from sporozoites
o Composed of large numbers of zoites
surrounded by a cyst wall
o Those that can be seen by the naked eye are
called macrocyst
o Microcysts are sarcocysts that remain about
the same width as the muscle fiber and can be
seen only under the microscope
Sarcocystis (Life cycle)
Definitive hosts are infected when they eat undercooked meat
w/c contains sarcocyst  sarcocyst wall digested  release of
zoites  enter intestinal cells and change into male and female
forms fertilization sporocysts passed out in the feces of the
host (definitive host)
Sporocysts ingested by intermediate hosts  sporozoites (w/c
are formed from sporocysts) penetrate intestinal epithelial
tissues hematogenous spreadbrain, muscles and other
tissues
** sporozoites enter muscle cells, and encyst and develop into
sarcocysts
** in other tissues, sporozoites become schizonts and then
encyst
Sarcocystis (Clinical Manifestations)
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▪In humans,
o Gastroenteritis, myalgia, weakness, mild
increase of creatine kinase
▪In the intermediate hosts, the brain, muscle, and kidney
tissues can be damaged
o Loss of appetite, fever, weight loss, and
anemia;
o Gait abnormalities, weakening of limbs, muscle
wasting, and head tilt
o Infected animals have been observed to move
in circles
▪Accidental ingestion of sporocysts results in direct infection
of humans without the intermediate host
o Occurs rarely and is called human
intramuscular sarcocystis
Sarcocystis (diagnosis)
▪In the definitive host, demonstration of sporocysts in the
fecal material using flotation method
▪In the intermediate host, detection of schizonts in the
skeletal muscle or in brain tissue during autopsy
▪Detection of antibodies in serum or CSF using Western blot,
IFA and ELISA
Sarcocystis (treatment and prognosis)
▪No effective treatment known
▪Corticosteroid – to decrease muscular inflammation
▪Trimethoprim-sulfamethoxazole – effective in treating
intestinal infections
▪In most animal infections due to this parasite do not cause
serious pathologic changes
Sarcocystis (Epidemiology)
▪Worldwide geographic distribution
▪Few human cases, most from SE Asia
▪Prevalence may be high due to numerous asymptomatic
infections
▪Other animals w/c may be infected: cattle, horses, dogs,
cats, rabbits, mice, and chicken
▪Infections in avian species have no known human threat
▪Sporocysts remain infective for many months in water at 4-
10C; infectivity prolonged during the cooler months of the
year
▪Infected carcasses left unburied on farmland can remain
infective for cats and dogs for weeks on cool weather
▪Freezing does not prevent spread
▪Macrocysts from sheep have to be heated for 20 mins to kill
them
Sarcocystis (prevention and control)
▪No known control methods
▪Uncooked dead animal should not be fed to other animals
▪Good sanitation and hygiene
 Parasitology – Coccidians by Dra Madrid
Cryptosporiosis
Cryptosporidium
▪Causal agent:
o Many species exist that infect humans and a
wide range of animals
o Although Cryptosporidium parvum and
Cryptosporidium hominis (formerly known as C.
parvum anthroponotic genotype or genotype 1)
are the most prevalent species causing disease
in humans, infections by C. felis, C.
meleagridis, C. canis, and C. muris have also
been reported
▪C. parvum – now considered a parasite of bovines w/c can
infect humans
▪C. hominis – infect only humans
▪Prevalence in the Philippines has been reported to be low at
2.6%
▪Coocidian protozoan
▪1907 – discovery of the organism
▪1976 – first case of human cryptosporidiosis
▪Infect several different hosts, can survive most
environments for long periods of time (“hard cysts”);
inhabits all climates and locales
▪Taxonomically classified as Sporozoa – oocyst releases 4
sporozoites
▪Monoxenous life cycle – completes entire cycle w/in a single
host
▪Sexual and asexual cycles
▪Six distinct developmental stages
Cyptosporidium (Life cycle)
▪6 distinct developmental stages:
o Excystation of the orally ingested oocyst in the
small bowel with the release of 4 sporozoites
o Invasion of intestinal epithelial cells; initiation
of asexual intracellular multiplication stage
o Differentiation of the microgametes (males)
and macrogametes (females)
o Fertilization initiating sexual replication
o Development of oocysts
o Formation of new, infectious sporozoite w/in
the oocyst, excreted in the stool
▪Sporulated oocysts, containing 4 sporozoites, are excreted
thru feces (and possibly respiratory secretions)
▪Following ingestion, excystation occurs sporozoites are
released and parasitize epithelial cells of GIT (or other
tissues) undergo asexual (schizogony or merogony) and
then sexual multiplication (gametogony)
▪Fertilization of macrogamonts (female) by microgametes
(male)  oocysts develop  sporulate in host, and excreted
o Because oocysts sporulate inside host,
autoinfection can occur
▪Oocysts are infective upon excretion, thus permitting direct
and immediate fecal-oral transmission
Cryptosporidiosis (Clinical Manifestations)
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Immunocompetent:
▪Acute self-limiting diarrheal illness
▪1-2 week duration
▪Frequent, watery diarrhea
▪N/V, abdominal cramps, low grade fever
Immunocompromised
▪Debilitating, cholera-like diarrhea (upto 20L/day)
▪Severe abdominal cramps
▪Malaise
▪Low grade fever
▪Weight loss
▪Anorexia
**Pa-add ng Cryptosporidosis epidemiology at yung persons
likely to be infected
Cryptosporidium: Transmission
▪Ingestion of fecally contaminated food and water
o Drinking water taken from surface water
sources such as lakes and river; swimming
pools and water park wave pools, untreated
water
o Cyst is resistant to various drinking water
filtration treatments such as chlorination
o Food can get contaminated; oocyst do not
survive cooking but can occur in foods not
heated or in foods not cooked after handling
▪Animal to person transmission
o From household pets is rare
o Definite correlation between calves shed
oocysts and pathogen present in 90% of all
dairy farms
▪Person to person
o Occurs at a high frequency in day-care centers;
clustered population
o Nosocomial settings
Cryptosporidium: Pathogenesis
▪Upon excystation, sporozoites released
o Adhere to the surface of the intestinal mucosa
o Sporozoite-specific lectin adherence factor:
agent of attachment to the intestinal surface
o Release of cytokines by mucosal cells 
activate phagocytes to release soluble factors
that increase intestinal secretion of water and
chloride; inhibit absorption
▪Epithelial cells damaged via:
o Direct result of parasite invasion
o Damage through T cell-mediated inflammation,
producing villus atrophy and crypt hyperplasia
 Parasitology – Coccidians by Dra Madrid
Cryptosporidium: diagnosis
▪Staining methods eg. Modified acid-fast stain to detect
oocysts in stool specimen
▪PCR
Cryptosporidium: treatment
▪No safe and effective therapy
▪Immunocompetent individuals – supportive care
▪Pharmacologic therapies:
o Immunocompromised patients, spiramycin and
dicalzuril sodium have produced partial
responses
o Paromomycin, a promising drug…decreases
severity of infection and improve intestinal
function
▪Immunologic therapies
o Feeding of bovine colostral immunoglobulin 
ameliorate symptoms; release of intestinal IgA
w/c clears infection
Cryptosporidium: prevention
▪Practice good hygiene
o Wash hands thoroughly
o Protect others: don’t go swimming if have
diarrhea
▪Avoid possibly contaminated water
o Do not swallow recreational water
o Do not drink untreated water (shallow wells,
lakes, rivers, ponds and streams)
o Do not use untreated ice or drinking water
during travel to areas with unsafe water supply
▪Avoid possibly contaminated foods
o Wash and/or peel raw vegetables and fruits
with clean water
o Avoid uncooked foods during travel to
countries with minimal water treatment and
sanitation systems
Cyclospora cayetanensis
▪Originally called a cyanobacterium-like body (CLB)
▪Life cycle presumed to be similar to other intestinal
coocidia
Cyclospora: parasite bology
▪The oocyst
o Formed after asexual and sexual cycles
o Passed in feces
o Are unsporulated when passed (after 5 or more
days, 2 sporocysts develop, each containing 2
sporozoites w/in oocyst)
▪Mature oocyst
o Sporulated
o Contains 2 sporozoites
o Considered infective stage
Page 4 of 7
o When ingested, sporozoites are released 
enter intestinal cells  undergo schizogony
and gametogony
** pa insert ng picture n life cycle, first slide sa page 10 ng trans
ng 2B
Cyclospora: Clinical Manifestations
** hindi ulit mabasa…ito yung secnd slide sa page 10
Cyclospora: diagnosis
▪Direct microscopic examination of fecal specimens using
concentration techniques and acid-fast staining (Kinyoun’
stain)
▪Fluorescent microscopy
o Appear blue or green circles
Cyclospora: treatment
▪Self-limiting disease
▪Treatment not necessary if symptoms are mild
▪The only effective drug is cotrimoxazole
▪No alternative drug if not tolerated
▪Oocyst disappear from the stools a few days after
treatment
Cyclospora: epidemiology
▪Epidemics reported: Haiti, Peru and Nepal
▪Raspberries from Guatemala were incriminated in infections
in the US
▪Leafy vegetables were found to contain oocysts Peru and
Nepal
▪Lettuce and basil pesto-salad incriminated in other cases in
the US
▪Contaminated water: main source of infection
▪No animal reservoirs found; cyclosporidiosis is considered a
human disease
Cyclospora: prevention and control
▪Follow good sanitary practices
▪In most endemic areas: boiling water seems to be the best
method since chlorination is not effective
▪Wash fruits and vegetables with clean water
Pneumocystis carinii
▪Causal agent
o P carinii is now considered a fungus based on
nucleic acid and biochemical analysis
 Parasitology – Coccidians by Dra Madrid
o However, it has morphologic and biologic
characteristics that, while incompletely
understood, place it close to the protozoa
▪Diffuse type of pneumonia in immunocompromised hosts
▪New name: Pneumocystis jiroveci
▪Exist in ______________________________ (hindi mabasa: 4
slide, page 11)
▪Painsert na rin ng slide 5 (picture naman to) at slide 6 sa
page 11
P carinii: structure
▪Cyst: thick walled; spherical to ovoid;
o 4-6 um in diameter; contains up to 8
pleomorphic sporozoites
▪Sprozoites: intracystic structure
▪Trophozoite: thin walled extracystic cell; represents
excysted sporozoite
▪Organism does not enter the host cell but attaches to its
surface during replicative cycle; no evidence of toxin
production
P carinii: clinical features
▪Dyspnea
▪Nonproductive cough
▪Fever
▪Diffuse infiltrates on CXR
▪Extrapulmonary lesions (< 3%):
o LN, spleen liver, and bone marrow
▪Illness may last from 4-6 weeks
▪Pneumocystis pneumonia (PCP) occurs in
immunosuppressed individuals and in premature
malnourished infants
P carinii: Pathogenesis
▪Portal of entry, not established
▪Likely mode of transmission: inhalation
▪In most individuals, dormant and dispersed in the lung eith
no apparent host response
P carinii: diagnosis
▪Id of organism in pulmonary tissue or lower airway fluids
through lung biopsy, inducement of sputum, bronchoalveolar
lavage, needle aspiration of lung
P carinii: treatment
▪Trimethoprim-sulfamethoxazole: DOC
▪Alternative drugs:
o Pentamidine
o Atovaquone
o Primaquine + clindamycin
o Trimthoprim + dapsone
Page 5 of 7
Toxoplasmosis
th
Toxoplasma gondii
▪Worldwide distribution
▪Infects humans and many species of animals
▪Definitive host: cat
o Complete cycle occurs in this host
o Cats get infected by ingestion of infective
oocyst from soil or tissue of infected animals
(mice)
o In cat intestines, undergoes schizogony,
gametogony and sporogony
T gondii (parasite biology)
▪In the cat
o In intestinal epithelium of the cat, merozoites
multiply (schizogony), followed by
differentiation into microgametocytes and
macrogametocytes (gametogony)
o Fertilization of the macrogamete gives rise to
an oocyst
o Oocyst passed out with cat feces in
unsporulated stage
▪Complete sporulation: 3-4 days
o In mature oocyst: 2 sporocysts, each with
sporozoites are formed
▪Can be ingested with contaminated food
or water by another host
o In humans or other hosts:
▪Mature oocyst excysts (inside intestine of
new host)  release 4 sporozoites 
penetrate lamina propria
▪Parasites enter lymphatics different
organs, tissues and body fluids
o Following entry of sporozoite into a new cell, t
transforms into a tachyzoite
▪Tachyzoites: formed during initial and
acute phase of infection
• Fast multiplying tachyzoites
give rise to slow multiplying
bradyzoites that form cysts
o Only these 2 stages are present in humans and
other animal intermediate hosts
o Asexual multiplication is by a variation of
binary fission called endodyogeny
▪Formation of plasma membrane by the 2
new daughter parasites even before the
division of the nucleus
o Cells in w/c endodyogeny occur eventually
burst releasing trophozoites w/c invade other
cells
o Pa-add ng toxoplasma life cycle (1st 2 slides sa
page 14)
T gondii: intermediate hosts
▪Man, rodents, farm animals
 Parasitology – Coccidians by Dra Madrid Page 6 of 7

o Does not have a well-defined wall

▪The cyst
T gondii: parasite biology o Usually contain bradyzoites
o Seen during chronic infections
▪Infective stages: o Well-formed wall
o Oocyst o Can be found in muscles and CNS
o Tachyzoites
o Bradyzoite
T gondii: clinical manifestations
▪The oocyst
o Ovoidal, thin walled
o Mature cyst in intestine of the new host  ▪Asymptomatic – majority of cases
excystsrelease 4 sporozoites, penetrates ▪Acute toxoplasmosis – fever, lymphadenopathy (infectious
intestinal mucosa; can also gain entry to the mononucleosis-like picture)
lymphatics and spread to other organs
▪Reactivation toxoplasmosis – in immunocompromised
▪The Tachyzoite patients (AIDS, cancer, drug immunosuppression, organ
o The transformed stage when sporozoite enters transplant recipients); presents with specific organ
a new cell involvement
o Found during initial and acute stage of o Encephalitis: most common manifestation
infection o Myocarditis, focal pneumonia
Rapidly multiplying

▪Choreoretinitis
 Present in humans and other animal o Occurs later in life in those who acquired
intermediate hosts
toxoplasma congenitally
 Can be transmitted to other humans o Retinal focal lesions: dec visual acuity, dec
by blood transfusion, transplacental
vision, retinal lesions on retinoscopy
and organ transplantation
▪The bradyzoite ▪Congential toxoplasmosis
o Mother to fetus
o Formed after tachyzoite
o Encysted form o Increased transmission rate in third trimester
o Found during chronic latent (asymptomatic) but increased severity of fetal disease in the 1st
infections trimester
o Persists for years in human tissue (brain, o Presents as hydrocephaly
retina, muscles, etc) o Hepatomegaly, cerebral calcifications, mental
o Can also be found in other animal intermediate retardation
hosts o Stillbirth and abortion may result when
o Can be transmitted by organ transplantation mothers acquire infection during the 1st
and eating meat of infected animals trimester of pregnancy

T gondii: diagnosis

T gondii: Multiplication ▪Observation of parasites in patient specimens s.c.

▪Sexual phase – occurs in cat
▪Asexual replication
o Occurs in both definitive and intermediate
hosts
o Endodyogeny – variation of binary fission
Formation of the plasma membrane

by the 2 new daughter parasites
before nuclear division
• Cells burst trophozoites
bronchoalveolar lavage for immunocompromised or lymph
node biopsy
▪Isolation of parasites from blood or other body fluids, by
intraperitoneal inoculate into mice or tissue culture
▪Detection of parasite gamete material by PCR, esp in
detecting congenital infection
▪Serologic test – routine method of diagnosis; detect
presence of antibodies against T gondii
o Sabin-Feldman methylene blue dye test
o IgM indirect fluorescent antibody technique
o IgM enzyme immunoassay

T gondii: parasite forms

T gondii: treatment
▪The trophozoite
o 4-8 um in length; 2-3 um in width ▪Regimen: pyrimethamine+sulfadiazine x 1month
o Crescent-shaped
o Pointed anterior end; rounded posterior end
▪Other drugs: clindamycin, azithromycin, clarithromycin,
dapsone, and atovaquone
o Rhoptries and micronemes found on the
anterior end; for cell penetration ▪Corticosteroids – prevent hypersensitivity reactions
o Nucleus: spherical on the posterior end ▪Trimethoprim-sulfamethoxazole – prophylaxis for
▪The pseudocyst immunocomromised
o Usually contains proliferating tachyzoites
o Can be taken in tissue sections of patients with
acute infections T gondii: Epidemiology
 Parasitology – Coccidians by Dra Madrid Page 7 of 7

▪Endemic worldwide both in humans and animals

▪Dse usually not manifested except in immunodeficiency or
immunosuppression
▪Infection more common in warm climates and at lower
altitudes than in cold climates and mountainous regions
▪Only 2% of human population is seropositive for T gondii

T gondii: prevention and control

▪Food should be protected from being contaminated with cat

feces
▪Cook eggs and meat well
▪Avoid unpasteurized milk.

END OF TRANS