COCCIDIANS The complexity in the life cycles of coccidians is a challenge

in terms of taxonomy
▪ Largest group of apicomplexan protozoa falling under
class Conoidasida CRYPTOSPORIDIUM
▪ Subclass of microscopic, spore-forming, single-celled C. parvum and C. hominis are the only spp. that can be
obligate intracellular protozoan seen in humans
▪ C. hominis is the most common based on DNA
COCCIDIA analysis
Members of Phylum Apicomplexa are provided with a
cluster of secretory organelles made up of: o Recognized in mice in 1907
o Rhoptries o Reported in humans in 1976
o Micronemes o Immunocompetent child
o Polar rings o Immunosuppressed adult
▪ Microtubules o Recognized globally in 1980s and 1990s
▪ Conoid o AIDS patient
o Outbreak among veterinary students
The secretion allows the parasite to enter the host cell
(epithelial cell of intestinal wall)

Cryptosporidium is a spore producing parasite found in

the intestine of infected people and animals
→ Parasites that inhabit the intestines are most likely
transmitted through fecal-oral route

o Cryptosporidium spp. is the most common

cause of Cryptosporidiosis.

1. Cryptosporidium = Cryptosporidiosis o There are many species of Cryptosporidium that

2. Cyclospora = Cyclosporiasis infect animals, some of which also infect humans.
3. Cystoisospora = Cystoisoporiasis
4. Sarcocystis =
o The parasite is protected by an outer shell that
5. Toxoplasma = Toxoplasmosis
allows it to survive outside the body for long
periods of time and makes it very tolerant to
COCCIDIOSIS
chlorine disinfection.
▪ Collective term for the disease caused by
coccidia
▪ Major problem in animal farming and zoo
management
▪ Among humans, they are opportunistic in
immunocompromised and immunodeficient
▪ These parasites are self-limiting; they may be
present in humans but it doesn’t cause harm
unless they are immunodeficient

LIFE CYCLE
▪ All coccidian follows the same life cycle
▪ There is an alteration of sexual and asexual
multiplication
o There are several species of Cryptosporidium that
It is typically characterized by THREE SEQUENTIAL are currently recognized.
STAGES:
1. Sexual cycle or Sporogony producing oocysts o It was initially reported that the only species that
2. Asexual Cycle or Schizogony (merogony) infect mammals was C. parvum and was believed
producing merozoites (meronts) to be the species infecting humans.
3. Gametogony resulting in the development of
male (micro) and female (macro) gametocytes o However, molecular tools, especially DNA analysis,
(gamonts). described the existence of another species,
 Cryptosporidium hominis found mainly in Oocyst → sporozoites → trophozoites → merozoites (micro
humans. and macro mates) → zygote → thin or thick-walled oocyst

DEFINITIVE HOST: not specific but able to infect

mammals especially humans

RSERVOIR: Calves, sheep, fish, birds and turkeys

MOT: fecal-oral-route

o All stages of development are completed in the

gastrointestinal tract of the [definitive] host.
- They do not have an intermediate host
Under electron microscope
CRYPTOSPORIDIUM LIFE CYCLE STAGES A. Sporozoites (in the brush area of epithelial cells
1. Oocysts will develop)
2. Sporozoites B. Trophozoites
3. Trophozoite C. Merozoites (producing)
4. Type 1 meront D. E. F. G. Gametes (and eventually into)
5. Type 2 meront H. Zygote
6. Microgamont
7. Macrogamont

CRYPTOSPORIDIUM OOCYST
▪ 4-5 micrometer wide
▪ One oocyst contains 4 fusiform sporozoites
▪ Does not stain with iodine and is acid-fast (requires
special stain for acid fast: Kinyoun stain, safranin
stain, etc.)
▪ Very hard and resistant (60C)
▪ Infective for 2-6 months in the environment
▪ Are released with fecal matter during onset of the
symptoms
▪ They are shed 5 days after infection
1. Oocysts are ingested through fecal-oral route.
→ means that it only takes 5 days to complete its
2. Then it travels to the epithelium of GI tract (their shell life cycle-from ingestion until it passes through
makes them resistant to stomach acids). the feces-which implies for a rapid infection)
3. Upon entering the cells, they will release 4 sporozoites
which will develop into trophozoites and further into INCUBATION PERIOD: between 1-14 days
merozoites (type I and II meront). ▪ Sequential application of zone and chlorine
4. These meronts eventually become Micro and Macro eliminate the cyst.
gamete-Microgamot, Macrogamot (gametogony)
and will mate in order to produce a zygote which will
develop into either a thick- or thin-walled oocyst.

a. The thin-walled oocyst can reenter the epithelial

cells creating an autoinfection.

b. The thick-walled oocyst on the other hand, can

pass through the feces.
▪ Unlike the thick-walled oocyst, the thin-walled
oocyst cannot survive the external
environment.
5. This cycle continues.
 CELL MEDIATED IMMUNE RESPONSE
▪ CDA + T cells
o Early infection
▪ CD8 + T cells
o Elimination
▪ CD154 and CD40
o Stimulate nitric oxide
o IFN-Y,IL-12
o T cell response
o Apoptosis
▪ Other cytokines
▪ TNF-a, IL-1B, IL-2, IL-4, IL-10, 1L-15, etc.
▪ Patient with AIDS
o Decreased CD4 + count
These are cells that help fight the parasite
CRYPTOSPORIDIUM HOMINIS HUMORAL IMMUNE RESPONSE
▪ Small cocci (2um-6um) These are antibodies produced
▪ Opportunistic parasite o IgM
▪ Zoonotic (cows, birds, reptiles, o IgG
fishes and humans) o IgA
▪ Causes short term infection in o X-linked immunodeficiency
humans o Mutations in CD14 gene
▪ Affects the host o Defected IgM cannot mount immune
gastrointestinal and response
respiratory epithelial cells DIAGNOSIS
Specimen Source:
Route of infection: fecal oral contamination
o Multiple stool specimens (3 specimens in order to
▪ Causes watery diarrhea
tell if negative)

PATHOGENESIS Diagnostic techniques

▪ Sporozoites adhere to the intestinal mucosa o Wet mount
▪ cells release cytokines o Modified Acid-Fast Stain
▪ increased intestinal secretion of sodium and chloride, o Direct Fluorescent Antibody (DFA) assay
water absorption is inhibited (malabsorption of water/
loss of water) Detection methods
▪ Epithelial cells damage by: ▪ Safranin stain
o parasite invasion and multiplication ▪ Trichrome stain
o T cells mediated villus atrophy (nasisira because of ▪ Enzyme immunoassay (EIA)
the parasites) ▪ Polymerase chain reaction (PCR)
▪ May produce up to 10-20 L watery stools per day ▪ Rapid immunochromatographic cartridge assays

Post Mortem Lesions

SYMPTOMS
o Gross lesions (not common)
▪ Appear 2-10 days after infection and last to 30 ▪ Hyperemia of intestinal mucosa. The mucosal
days folds are markedly thickened, and there are
▪ Diarrhea numerous pinpoint foci of hyperemia.
▪ Stomach cramps o Microscopic lesions
▪ Dehydration ▪ Mild to severe villous atrophy
▪ Nausea ▪ Spherical organisms in the brush border
▪ Vomiting
▪ Fever ❑ Histopathology
▪ Weight loss - Epithelial cell
▪ Sometimes no symptoms (persons that are not
immunocompromised)

INNATE IMMUNE RESPONSE

▪ White blood cells phagocytize parasites

o Segmented neutrophils
o Macrophages
o Lymphocytes
o Eosinophils
Under bright field microscopy
Needs further analysis since they look just like RBC and
air bubbles
Should do differential interference contrast or phase
contrast

Positive for C. parvum

Use dark field microscope TREATMENT

o Nitazoxande
o Paromomycin
o Azithroycin
o Individuals with AIDS
o Anti-retroviral therapy
PREVENTION
▪ Boiling and microfiltration of drinking water
▪ Microfiltration removes oocyst from the water
supply
▪ Low levels of chlorine does not kill cysts
o C. parvum 240,000 times resistant to
chlorination than Gardia
o Chlorine dioxide – ineffective for oocyst

WATERBORNE PREVENTION
o Do not swallow recreational water
o Lakes, rivers, streams, untested wells
 o Do not drink untreated water
o Travelers and hikers
o Boil water for 15 mins or use filter rated
for “cyst removal’
o Don’t rely on chemical treatments
o Do not swim with GI infection
FOODBORNE PREVENTION
o Wash vegetables with detergent soap
o Proper human/animal waste disposal
o No bare hand contact or ready-to-eat foods
o No food workers with GI illness
o Until 2 weeks after end of diarrhea
o Handwashing

PATHOGENESIS AND CLINICAL MANIFESTATION

Pathology: Cyclosporiasis
o Initial symptoms include malaise and low-grade
fever
o Chronic and intermittent watery diarrhea
o fatigue, anorexia, weight loss, nausea, vomiting,
abdominal pain, flatulence, bloating, and dyspnea
o D-xylose malabsorption
o Infections are usually self-limiting and immunity
may result with repeated infections

DIAGNOSIS
o Direct microscopic examination of fecal smears
under high magnification (400x)
CYCLOSPORA CAYETANENSIS - Looks like oil-droplet, Well defined
o Cyclospora cayetanensis was originally called a membranes
cyanobacterium-like body (CLB) but upon careful - Use phase contrast microscope to
study, it was found to be a coccidian parasite. differentiate with fat globules or air bubbles
-

LIFE CYCLE
o ingestion of sporulated oocyst, which contains two
sporocysts with two sporozoites each.
o sporozoites invade the epithelial cells of the small
intestines
o Multiple fissions of these sporozoites take place o Various concentration techniques and acid-fast
inside the cells to produce meronts, which contain 8 staining (kinyoun’s stain) are also useful.
to 12 merozoites during the first generation, and
only four merozoites in the second generation.
o Some of the merozoites develop into male (micro)
and female (macro) gametes.
o The microgametes fertilize the macrogametes to
produce oocysts, which are passed out with feces
when the host cells are sloughed off from the
intestinal wall.
o The oocysts undergo complete sporulation within
7 to 12 days in a warm environment
o Oocyts are autofluorescent under fluorescent CYSTOISOSPORA BELLI
microscopy they appear as blue or green circles o This is the causative agent of a medical condition
depending on the filter (365-450 DM) affecting the small bowel called Cystoisoporiasis.
- This technique is useful for screening of o The other known species Isospora hominis is now
Cayatenensis taxonomically grouped under genus Sarcocystis.

o UV microscopy is also used

o The sporulated oocyst contains two sporocysts each

containing four sporozoites (infective stage).
o Safranin staining and microwave heating are also o When ingested via contaminated water or food, the
helpful sporozoites excyst in the small intestine releasing
sporozoites, which penetrate the epithelial cells, thus
starting the asexual stage or the schizogonic phase
of the life cycle
o Oocysts are thin walled, transparent, and ovoid in
shape.
o They appear as translucent, oval structures measuring
20 to 33 pm by 10 to 19 µm

o A polymerase chain reaction (PCR) technique has

been developed to differentiate Cyclospora from
closely related Eimeria species.

LIFE CYCLE
o The sporozoites develop in the epithelial cell to form a
schizont, which ruptures the host epithelial cell
liberating merozoites into the lumen.
o These merozoites will then infect new epithelial cells
and the process of asexual reproduction in the intestine
continues.
o This process may continue for weeks or months. Some
of the merozoites undergo gametogony to produce
macrogametes and microgametes (sexual stages),
which fuse to form a zygote that eventually matures to
form an unsporulated oocyst.
o Sporulation usually occurs within 48 hours after o Oocyst can be seen in a fecal smear stained by a
passage with the stool. modified Ziehl-Neelsen method, where they stain
granular red color against a green background

o Phenol-auramine, as well as iodine staining of the

specimen can help visualize the organism.

Phenol-auramine - is a differential stain, that can be seen

PATHOGENESIS AND CLINICAL MANIFESTATION under dark field microscopy
o Self-limiting gastroenteritis (immunocompetent)
o Severe diarrhea and malabsorption (severe infections)
o Low grade fever, anorexia, vomiting, general body
malaise, anorexia, weight loss, and flatulence
o Stools usually contain undigested food, mucus, and
Charcot-Leyden crystals
o Immunocompromised
o May cause severe diarrheal illness
o Mucosal bowel biopsy may show flattened o Acid-fast stain, such as Kinyoun’s stain or an
mucosa and damaged villi auramine rhodamine stain, is also useful
o Infiltration of the lamina propria with
lymphocytes, plasma cells, and eosinophils has
been reported

DIAGNOSIS
o The oocysts of c. belli may be detected in the feces
by:
❑ direct microscopy or formalin ether/ethyl o String capsule (enterotest) and duodenal aspirate
acetate concentration examinations may be of value
❑ zinc sulfate and sugar flotation.
When should the test be performed:
▪ When a physician suspects a parasitic infection,
but no parasites were found in stool sample.
▪ As its sensitivity is comparable to duodenal
aspirate, it eliminates the need of duodenal
intubation.
 TOXOPLASMA GONDII o These oocysts will travel into the epithelial cells
Unique from other kinds of coccidia because it has an of intestinal mucosa where it will complete its
intermediate host cycle (asexual or schizogony; sporogony’ and
o Toxoplasma is an intracellular parasite, which gametogony)
infects different kinds of nucleated cells including
macrophages

DEFINITIVE HOST: members of Felidae (domestic cats

and their relatives)

LIFE CYCLE: It follows a typical coccidian life cycle

T. GONDII TACHYZOITES
→ consisting of schizogony, gametogony, and
sporogony in the intestinal epithelium. o Rapidly growing stage observed in the early stage
→ The extraintestinal stages are the asexual stages: of infection (acute phase) habits in the body fluid
tachyzoites and bradyzoites. o Crescent shaped (4-8pm in length and 2-3pm in
width)
Since there is an extraintestinal stage, it can be transmitted o Organelles, such as rhoptries and micronemes,
not only via fecal-oral route. which are associated with cell penetration, are found
in a short conoid on the anterior end.
o A spherical nucleus is found in the posterior end
o ASEXUAL FORM
o Multiplies by binary fission (endodyogeny)
▪ Endodyogeny - can replicate or multiply
even in the absence of the nucleus [kahit
hindi pa buo ang nucleus]
o It can infect phagocytic and nonphagocytic cells

T. GONDII OOCYST
a. UNSPORULATED OOCYST
o Non infectious
o Subspherical to spherical
o Takes 1-5 days to sporulate
o Requires oxygen to sporulate
T. GONDII BRADYZOITES
o Slow growing stage inside tissue cyst
o Marks the chronic stage of infection
o Resistant to low pH and digestive enzymes during
stomach passage
o Protective cyst wall is finally dissolved and
bradyzoites infect the tissue and transform into
tachyzoites
b. SPORULATED OOCYST o Bradyzoites are released in the intestine and are
o 10u diameter highly infective if ingested
o Subspherical to ellipsoidal
o Each has ellipsoidal sporocyst
o Each sporocyst contains 4 sporozoites
o Infective (remain for months)

Bradyzoites are attached to the muscle tissue. When this

tissue is ingested (raw meat) it will survive the stomach acid
and therefore can continue its life cycle in the intestine. This
becomes now the infective stage.
Bradyzoites will transform into tachyzoites which is the fast- immune system rather than as a response to an
growing stage. acute infection.
o Among the immunocompromised patients, the most
The reason why it is called chronic stage is because it can common manifestation is encephalitis,
stay in the muscle tissue for a long period of time without myocarditis and focal pneumonia have also been
manifesting any signs and symptoms reported

LIFE CYCLE PATHOGENESIS

❑ Acquired taxoplasmosis (mild lymphatic
inflammation)

❑ Congenital toxoplasmosis
o Intracerebral calcification
The extraintestinal stage: bradyzoites and tachyzoites o Chorioretinitis
o Hydrocephaly
Maglalabas ng feces yung cat (Felidae) containing the o Microcephaly
oocyst. This fecal contaminated material can be ingested by o Convulsions
different animals including humans. Pag naingest nila o Mental retardation
pwede magtransform into tachyzoites tas pwede pumunta o Cardiomegaly
sa muscle tissue. Yung Felidae cat can ingest this tissue
containing the bradyzoite and then the cycle goes on.

Pag naingest yung bradyzoites sa intestine ng definitive

host, maliliberate siya and magiging tachyzoites and then
into trophozoites, merozoite, gametocyte and then the
process continue.

2 ways of infection: through ingestion of tissues or fecal

contaminated material LABORATORY DIAGNOSIS

TRANSMISSION 1. Microscopy
o Contaminated water or food by oocysts - Stains: giemsa, PAS, GMS
o Ingestion of tachyzoites and bradyzoites in the 2. Serodiagnosis
flesh of infected host. - Antibody detection
o Undercooked meat - Antigen detection
o Mother of fetus 3. Molecular diagnosis
o Organ transplant (rare) - PCR
o Blood transfusion (rare) 4. Imaging
PATHOGENESIS AND CLINICAL MANIFESTATIONS - MRI & CT scan
- USG – for congenital toxoplasmosis
o Toxoplasmosis is commonly asymptomatic as long 5. Others: Animal inoculation, Skin test of
as the immune system of the patient is functioning Frenkel
well.
o Cysts can be found in the brain, skeletal and heart
muscles, and retina.
o Clinical manifestations become apparent when the
immune system is suppressed as in old age, drug-
induced immunosuppression after organ
transplantation, or in the case of AIDS.
o More often, symptoms appear when there is relapse
of chronic infections as a result of a suppressed
When we do diagnosis hindi pwedeng isa lang, we need to
use different test
 SARCOCYSTIS SPP. to 19 um by 8 to 10 um, and contain four sporozoites
and a discrete refractile residual body.
o Sarcocystis is a genus of intracellular protozoa
o Sporocysts are capable of surviving on the
reported to infect humans and animals worldwide
ground and infecting intermediate hosts
o Infection with this parasite is known as
Sarcosporidiosis or Sarcocystosis.

Sarcocystis Sarcocystis
hominis suihominis
Intermediate Cattle Swine
host
Definitive Human Human
host
Sporocyst Bigger Smaller
Effect Intestinal sarcocystosis
Wet mount
HISTORY LIFE CYCLE
Same as T. Gondii, but the definitive Host are the humans
o This parasite was first reported in 1843 by Miescher
and the intermediate host are the cattle and swine
as white threadlike cysts in striated muscles of a
house mouse.
o It was simply referred to as Miescher's tubules until
1899, when the name Sarcocystis miescheriana was
proposed to identify the said parasite.
o There are about 130 recognized species under
Sarcocystis including S. hominis and S. suihominis
(humans definitive hosts)

SARCOCYTIC FORMS - ZOITES

▪ It is a banana shaped cell, with a pointed anterior
end, also known as the apical complex, which
possesses micronemes, micropores, and rhoptries,
and believed to be associated with host cell
penetration and creation of an intracellular
environment suitable for parasite growth and
development

PATHOLOGY AND CLINICAL MANIFESTATION

▪ So ito na yung sporozoites which will penetrate the
▪ Rare invasive form
intestinal mucosa
(lymph nodes, muscles,
SARCOCYSTIS FORMS- OOCYST
and the larynx)
▪ Sporulated oocysts and individual sporocysts can be
passed out in the feces of an infected definitive host. Pathology: Vasculitis and
The sporulated oocyst undergoes sporogony creating Myositis
two sporocysts.
▪ Once sporogony is complete, the oocyst itself ▪ Sarcocystosis has also
undergoes lysis, releasing the sporocysts into the been associated with
environment. Sporocysts of most species measure 15 acute fever, myalgias,
bronchospasm, pruritic
 rashes, lymphadenopathy, subcutaneous nodules with few as 50 sporocysts from fecal samples that had
concurrent eosinophilia, elevated erythrocyte been stored in potassium dichromate (K2 Cr2 07)
sedimentation rate, and elevated creatine kinase levels. for as long as 6 years.

Causes increase ESR

(inflammation)

▪ Intestinal form - nausea,

abdominal pain, and diarrhea

DIAGNOSIS
1. Stool examination
▪ Fecal flotation (formalin ether/ethyl acetate
and other sedimentation methods)

Wet mount:

Under Differential interference contrast

2. Biopsy of infected muscle

o Sarcocysts of:
➢ S. hominis are microscopic in muscles of
cattle, whereas those of
➢ S. suihominis are macroscopic in muscles of
swine.
o Sarcocysts are identifiable with hematoxylin and
eosin stain.
o Confirmatory staining with the periodic acid-
Schiff (PAS) can be performed as the walls stain
positively

3. Polymerase Chain Reaction

o polymerase chain reaction (PCR) amplification of
the 18S rRNA was demonstrated to be useful in
distinguishing S. hominis, S. fusiformis, and S.
cruzi sarcocysts and oocysts.
o The technique makes possible amplification and
identification of species’ specific gene
sequences based on DNA extracted from as few
as seven excreted sporocysts (the equivalent of
3% oocysts) from freshly prepared material, or as