Debre Berhan University College of medicine Dep MLS Medical parasitology 2012/2020

5. Apicomplexa (sporozoa)
5.1General future and classification
The Apicomplexa (also called Apicomplexia) are a large phylum of parasitic alveolates.
Most of them possess a unique form of organelle that comprises a type of plastid called
an apicoplast, and an apical complex structure. The organelle is an adaptation that the
apicomplexan applies in penetration of a host cell.
The name of the taxon Apicomplexa derives from two Latin words—apex (top)
and complexus (infolds)—and refers to a set of organelles in the sporozoite.
It includes

• 1. Intestinal coccidian
– Cryptosporidium
– Isospora
– Cyclospora
• 2. Blood and tissue coccidian
– Plasmodium
– Toxoplasma
– Babesia
– Sarcocystis

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5.2 Intestinal coccidian

Cryptosporidium, Isospora, Cyclospora
General characteristics
 Considered as opportunistic parasite in immunocompromized person.
 Complete entire life cycle in single host.
 Within the intestinal epithelial cells of the host.
 Characterized by a thick walled oocyst excreted in faeces
 Are transmitted by the fecal-oral route
5.2.1 Cryptosporidium

History

 Initial identification in 1907 several Cryptosporidium species have been

identified.in a wide variety of animals ranging from fish to humans.
 The first human cases of cryptosporidiosis were reported in 1976
 During the 1980’s Cryptosporidium was recognized as a major cause of diarrhea
in AIDS patients and often resulted in death.
 However, it is now recognized that Cryptosporidium is a common cause of
diarrhea in immunocompetent persons and has probably been a human
pathogen since the beginning of humanity.
 Two species infecting humans have been identified: C. parvum and C. hominis.

 C. hominis & C. parvum is species mostly causing disease in humans

 C. hominis infects only humans but C. parvum also infects many other mammals
 C. felis, C. meleagridis, C. canis, and C. muris infections have also been reported
Transmission
• Mainly through the ingestion and possibly inhalation of sporulated oocysts mainly
through contaminated water & Occasionally food sources, such as chicken salad
• Anthroponotic (human to human) transmission
• Autoinfection
• zoonotic transmission

Life cycle

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1. Following ingestion (and possibly inhalation) by a suitable host excystation

occurs. The sporozoites are released and parasitize epithelial cells of the
gastrointestinal tract undergo asexual multiplication (schizogony or merogony) and
then sexual multiplication (gametogony) producing microgamonts (male) and
macrogamonts (female).

2. Upon fertilization of the macrogamonts by the microgametes , oocysts develop that

sporulate in the infected host. Two different types of oocysts are produced, the thick-

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Debre Berhan University College of medicine Dep MLS Medical parasitology 2012/2020

walled, which is commonly excreted from the host, and the thin-walled oocyst, which is
primarily involved in autoinfection.

3. Oocysts are infective upon excretion, thus permitting direct and immediate fecal-oral
transmission.

Note: that oocysts of Cyclospora cayetanensis and Isospora another important

coccidian parasite, are unsporulated at the time of excretion and do not become
infective until sporulation is completed.

Clinical Features

 Vary from asymptomatic to severe, life-threatening illness;

 infection usually self-limiting diarrhea of 1-2 weeks duration charactrized by
copious watery diarrhea, vomiting, intense abdominal pain, anorexia and weakness
 among immunocompromised patient (AIDS)  develops severe chronic diarrhea
which may last for months  malabsorption
 Incubation period ~ 7 days ( 2 to 10 days).
 In immunocompetent persons, Symptoms are usually short lived (1 to 2 weeks)
 Persons with AIDS (CD4 counts <200/µl)
• Chronic and more severe lasting for months or even years
 Small intestine- most commonly affected.
 The lungs, and possibly conjunctiva may be affected.

5.2.2 Cyclospora
Specie: Cyclospora cayetanesis

 1986 cases of prolonged watery diarrhea among immunocompromised (AIDS)

patient has been reported worldwide .
 Species Name C.Cayetanensis was given in 1993 Initially called ‘cyano-bacteria like
body’ (CLB) or large cryptosporidium

Clinical Presentation

After an average incubation period of one week, symptomatic infections typically manifest
as watery diarrhea of varying severity. Other manifestations include complications of
dysentery, further abdominal symptoms, and sometimes non-specific systemic symptoms
(e.g. headache, low-grade fever). Untreated infections typically last for 10–12 weeks and

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Debre Berhan University College of medicine Dep MLS Medical parasitology 2012/2020

may follow a relapsing course. The duration of symptoms and associated weight loss are
greater in individuals with HIV or possibly other immunosuppressive conditions. Infections
can be asymptomatic in disease-endemic regions.

5.2.3 Isospora(Cyclospora)
Isospora belli
 Life cycle similar to Cryptosporodium except that oocysts mature in environment

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Clinical feature
• Disease: Isosporosis/Intestinal coccidiosis
• often asymptomatic (seldom reported)
• symptoms range from mild gastro-intestinal distress to severe dysentery
• often self-limiting, but can become chronic (wasting, anorexia) symptoms more
severe in AIDS patients

5.2.4 Sarcocystic species

Sarcocystis is a genus of parasites, the majority of species infecting mammals, and some
infecting reptiles and birds. There are 130 species. The definitive host usually does not
show any symptoms of infection, but the intermediate host does.

Sarcocystis hominis and S. suihominis use humans as definitive hosts and are responsible for
intestinal sarcocystosis in the human host. Humans may also become dead-end hosts for
non-human Sarcocystis spp. after the accidental ingestion of oocysts.

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1. Both sporulated oocysts (containing two sporocysts) and individual sporocysts can
be passed in stool .
2. Sporocysts contain four sporozoites and a refractile residual body. Sporocysts
ingested by the intermediate host (cattle for S. hominis and pigs for S. suihominis)
rupture, releasing sporozoites. Sporozoites enter endothelial cells of blood vessels
and undergo schizogony, resulting in first-generation schizonts. Merozoites derived
from the first-generation invade small capillaries and blood vessels, becoming
second-generation schizonts. The second generation merozoites invade muscle cells
and develop into sarcocysts containing bradyzoites, which are the infective stage for
the definitive host .
3. Humans become infected when they eat undercooked meat containing these
sarcocysts. Bradyzoites are released from ruptured cysts in the small intestine and
invade the lamina propria of the intestinal epithelium .
4. There, they differentiate into macro- and microgametocytes. Fusion of male and
female gametes results in the formation of oocysts .
5. Oocysts sporulate in the intestinal epithelium and are shed from the host in feces.
6. Due to the fragile nature of the oocyst wall, individual sporocysts may also be
detected in feces.
Geographic Distribution

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Worldwide, but more common in areas where livestock is raised.

Clinical Presentation
In cases of intestinal sarcocystosis, when humans serve as the definitive hosts, infections
are often asymptomatic and clear spontaneously. Occasionally, mild fever, diarrhea, chills,
vomiting and respiratory problems may occur. When humans become infected with
sarcocysts of non-human species, the infections are not intestinal but rather result in
muscle cysts; symptoms such as myalgia, muscle weakness and transitory edema may
occur. In these cases, humans are dead-end intermediate hosts.
Laboratory Diagnosis
For intestinal sarcocystosis caused by S. hominis and S. suihominis, diagnosis is made by the
observation of oocysts or sporocysts in stool. They are easily overlooked as they are often
shed in small numbers. Also, the two species cannot be separated by oocyst or sporocyst
morphology. When humans serve as dead-end hosts for non-human Sarcocystis spp.,
diagnosis is made by the finding of sarcocysts in tissue specimens.

Laboratory Diagnosisof Intestinal coccidia

 Macroscopic
 Modified acid-fast staining technique
 Immunological assay

Modified Acid-Fast Staining Procedure

Oocysts of cryptosporidia, cyclospora, isospora, and sarcocystis are acid fast and will be
detecte.

A blue-green background, or contrasting counterstain, of fecal debris allows the oocysts to

stand out. The oocysts are variably stained: some will stain light pink to deep purple, while
others may be unstained. The oocysts (8 to 10 μm) may not be perfectly round; some may
appear collapsed or distorted on one side.

. Unlike the Ziehl-Neelsen Modified Acid-Fast Stain, this stain does not require the
heating of reagents for staining.

Specimen:

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Debre Berhan University College of medicine Dep MLS Medical parasitology 2012/2020

Concentrated sediment of fresh or formalin-preserved stool may be used. Other types of

clinical specimens such as duodenal fluid, bile, pulmonary samples (induced sputum,
bronchial wash, biopsies) may also be stained.

Reagents:
There are four steps to this procedure requiring the following solutions:
1. Absolute Methanol
2. Acid Alcohol: 10 ml Sulfuric Acid + 90 ml Absolute ethanol. Store at room temperature.
3. Kinyoun’s Carbol fuchsin: may be purchased commercially.
4. 3% Malachite green: dissolve 3 g of malachite green in 100 ml of distilled water. Store
at room temperature.

Procedure:

1. Prepare a smear with 1 to 2 drops of specimen on the slide and dry on a slide warmer
at 60°C until dry. Do not make the smears too thick!
2. Fix with absolute methanol for 30 seconds.
3. Stain with Kinyoun’s carbol fuchsin for one minute. Rinse briefly with distilled water
and drain.
4. Destain with acid alcohol for 2 minutes. Rinse with distilled water and drain.
5. Counterstain with Malachite green for 2 minutes. Rinse briefly with distilled water and
drain.
6. Dry on a slide warmer at 60°C for about 5 minutes. Mount with a coverslip using
desired mounting media.
7. Examine 200 to 300 fields using 40× or higher objectives. To confirm internal
morphology, use 100× oil immersion objective.

Quality Control:
A control slide of Cryptosporidium spp. from a 10% formalin preserved specimen should be
included with each staining run. Cryptosporidium spp. stains a pinkish-red color. The
background should stain uniformly green.
(Sourcehttps://www.cdc.gov/dpdx/diagnosticProcedures/stool/staining.html#Acid-Fast)

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Debre Berhan University College of medicine Dep MLS Medical parasitology 2012/2020

Prevention and control: Personal hygiene and avoid eating of raw unwashed food.
Read current treatment method.

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