THE APICOMPLEXAN
Overview of classification of parasites
Kingdom- Animalia
Subkingdom Protozoa Subkingdom Metazoa
phylum sarcomastigophora phylum nematoda
phylum sporozoa phylum platyhelminth
phylum ciliophora
phylum microspora
Kingdom phylum class order family genus
Characteristics of apicomplexa
are microscopic, spore-forming intracellular Single-
celled organisms that multiply in human host
have complex life cycle, well-developed asexual &
sexual stages.
motility is absent in most cells except male gametes.
Apicomplexa....
Sporozoans produce special spore like cells called
sporozoites ff sexual reproduction w/c are important in
transmission of infections;
most form thick-walled zygotes called oocysts;
Reproduction
asexually by schizogony (merogony)
sexually by sporogony
Two groups
Blood and tissue coccidia:
Plasmodium species & Babestia spp.
Toxoplasma gondii
Intestinal coccidia:
Isospora belli
Cryptosporidium parvum
Cyclospora cayetanensis
most coccidia are particularly important as pathogens
in immunosuppressive conditions
Toxoplasmosis
Toxoplasmosis
Toxoplasmosis
Introduction
Toxoplasma gondii is an animal coccidian
parasite which causes toxoplasmosis
congenital toxoplasmosis the most serious
form of human infection.
T. gondii has a worldwide distribution.
Toxoplasma gondii is one of the most well studied
parasites because of its medical and veterinary
importance.
1908-Nicolle and Manceaux
It is used extensively as a model for cell biology of
apicomplexan organisms.
advantages of using T. gondii for cell biology
large enough to be easily seen under the
light microscope
it can be grown in virtually any warm-
blooded cell Line
only one species that infects all hosts
Toxoplasmosis…
Incidence
varies greatly by country and by age
1/3 of the global population
congenital toxoplasmosis:1-10 /10,000 live births
Immunocompromised individuals are at risk for
reactivation of latent infection, including potentially
fatal encephalitis.
Causes and Risk Factors
The house cat serves as definitive hosts
Routes of transmission
• accidental ingestion of oocysts passed in cat
• ingesting tissue cysts
raw or undercooked meat (lamb, pork and beef )
drinking unpasteurized milk
contaminated water, or unwashed fruits or
vegetables
• direct transmission of tachyzoites from mother to fetus
(congenital infection)
• blood transfusion /organ transplantation
Morphology
Tachyzoite-the
intracellular parasites,
3x6µm in size
Crescent or oval in
shaped, one end is
rounded and the other
end is pointed
Tachyzoites are the
actively proliferating
trophozoites, which are
observed during the acute
stage of infection
morphology…
Tissue cyst- filled with
bradyzoites
Bradyzoites crescent-shaped
cell and 7 x 1.5 μm measures
Bradyzoites in the chronic
stage of infection, develops
slowly and multiplies in the
tissues
Tissue cysts are found most
commonly in the brain and
in skeletal and cardiac
muscle
Cyst measures 10-100m
Morphology…
Oocysts:
The oocyst contains
two sporocysts, each
of which contain four
sporozoites.
10 µm in diameter
Life cycle
complex
It involves three distinct inter-linked cycles
1. feline cycle
associate with formation of oocyst
oocyst can infect DH and IH
liberate sporozoites on reaching the intestinal
of cats
sporozoites penetrate the intestinal mucosal
cells
Life cycle…
Sporozoite multiply asexually(endopolygeny)
to form endozoites
After some cycle of multiplication,
gametogony is initiated → zygote is formed
→ covert to oocyst
Life cycle…
2. Feline- non feline cycle
IH infected by oocysts through contaminated
food and water
Liberate sporozoites on reaching the intestinal
of IH and multiply asexually to form endozoites
All cells except RBC can be infected
the infected cell rupture to release endozoites
Endozoites continue to multiply in different
sites
Endozoite convert to cystozoites that are
3. Non feline non feline cycle
Cystozoites in the tissue cysts of non-feline
animals are infective to other animals or
birds
Only asexaul multiplication takes place
Cystozoites infect the intestinal cells of non-
feline and intiate endopolgeny
They infect deeper tissues to form tissue
cysts containing cystozoite
Cat ingests tissue cysts containing bradyzoites
Gametocytes develop in the intestine
Oocysts appear in the cat’s feces 3-5 days after
infection by cysts
Oocysts require oxygen and they sporulate in 2-
3 days
Epidemiology and transmission
Transmission to humans
Oral
Ingestion of under cooked Pork or Lamb
meat –tissue cyst.
Exposure to oocysts
Ingestion of contaminated food and water
Direct contact with cat feces.
Others
Transplacental (Vertical transmission)
Blood Product Transfusion
Organ Transplantation
The life cycle of Toxoplasma gondii begins
when a cat ingests toxoplasma-infected tissue
from an intermediate host(rodent)
Tissue cysts within the muscle fibers or brain
are digested in the cat’s digestive tract.
The parasite multiplies in the intestine of the cat
(sexual development) is eventually shed in cat
feces
Epidemiology:
Human infection:
Ingesting raw meat
Sporulated oocysts from contaminated soil
Birds & rodents are important in picking oocysts from soil or
scavenging bradizoites from infected animals
WW distribution
Hot, dry climates-lower incidence
Temperate, humid climates-high incidence
Pathology
Toxoplasma gondii invades numerous organs,
infecting a broad spectrum of cell types.
Tachyzoites→macrophages→blood→organs(invasion
,asexual multiplication) → cell death.
The resulting necrosis attracts inflammatory host
cells(lymphocytes and monocytes) → pathology.
As host resistance develops, tissue cysts may form in
many organs(brain and muscle)-quiescent cysts
evade the adaptive host immune
tissue cysts periodically rupture, the released
bradyzoites are killed by the host immune system.
If immunocompromised:bradyzoites develop
into tachyzoites- active toxoplasmosis
3 categories
a. Postnatally acquired toxoplasmosis
b. Immunocompromised related
toxoplasmosis
c. Congenital toxoplasmosis
Postnatally acquired toxoplasmosis
> 80-90% of primary infections- asymptomatic
Mostly recognized finding
cervical lymphadenopathy accompanied by low-grade
fever
mononucleosis-like syndrome(Epstein-Barr virus)
fever
swelling of the lymph nodes
hepatosplenomegally
myalgia
Atypical lymphocytosis in the blood
Enlarged cervical lymph nodes (arrows) in a 19-year-old boy
with acquired toxoplasmosis
Sever disease
retinitis, which may result in strabismus(cross-eye) or
blindness.
A characteristic residual pigmented scar is left on the
retina after resolution of infection.
immunocompromised adults
Brain lesions
fever, headache, confusion & seizures
Systemic manifestations
Myocarditis & pneumonitis
Immunocompromised toxoplasmosis
Cerebral manifestations
Altered mental status
Focal Neurological deficit
motor weakness
speech disturbances
cranial nerve palsy
movement disorders
visual defects
sensory ,cerebelar dysfunction
Congenital toxoplasmosis
Congenital toxoplasmosis
mother infected before conception-no congenital
toxoplasmosis
new infections with detectable maternal parasitemia
~50% transmission rate of congenital toxoplasmosis
transmission rate and severity of disease in the fetus are
inversely proportional
mothers who develop acute toxoplasmosis in the first
trimester have a much lower fetal transmission rate
fetuses exposed early are at much higher risk for severe
symptoms or death and spontaneous abortion
Gestational age at Risk of congenital Development of
maternal infection(95% CI) clinical signs in
seroconversion weeks % infected
offsprings(95% CI)%
13 6(3-9) (34-85)
26 40(33-47) 25(18-33)
36 72(60-81) 9(4-17)
Table 1. Risk of Toxoplasma gondii congenital infection
(transmission) and development of clinical signs in offspring
before age 3 years, according to gestational age at maternal
seroconversion
Two types:
• Asymptomatic (inapparent) congenital toxo
60% of infected
may suffer from Long Term Sequelae
~90 percent of infected babies appear normal
at birth
80 to 90 percent will develop sight-threatening
eye infections months to years after birth
~10 percent will develop hearing loss and/or
learning disabilities
Symptomatic Congenital toxoplasmosis
40% of infected
About one in 10 infected babies has a severe
Toxoplasma infection that is evident at birth
severe eye infections, an enlarged liver and
spleen, jaundice
Some die within a few days of birth
some suffer from mental retardation, severely
impaired eyesight and seizures
Severe damage to fetus = stillbirth or abortion
Hydrocephalus with bulging forehead (left) and
microophothalmia on the left eye
Hydrocephalus
• rare symptom- increases in the volume of cerebrospinal fluid
Figure 1.Hydrocephalic and spastic baby
(CSF) within the ventricles of the brain as a result of host
reaction to the parasite
Lab. Diagnosis
Morphologic Dx
tachyzoites in
circulating WBC
bone marrow
lung
spleen
Histopathologic examination
Culture or animal inoculation (rare)
Serologic (mainly)
Molecular Dx-
Detection of T. gondii parasites in c.s.f.
• The c.s.f. usually contains neutrophils and
mononuclear cells.
Method
1. Centrifuge the c.s.f. in a conical tube at medium
to high speed(about 1 000 g for 5 minutes).
o Transfer the supernatant fluid to another tube.
This can be used for protein testing (raised in
Toxoplasma encephalitis).
2. Tap the bottom of the tube to mix the sediment.
Spread a drop on a slide and allow the smear to air-
dry. Fix with methanol for 1–2 minutes.
3. Stain the smear using Giemsa technique
4. When dry examine the preparation microscopically
40X objective to detect the parasites & 100X
objective to identify the tachyzoites.
Serological diagnosis of toxoplasmosis
A. Sabin-Feldman dye test: the most reliable test for
the diagnosis of acute toxoplasmosis in pregnancy,
and to test neonates
• is highly sensitive and specific test
• It is performed in Reference Laboratories
B. The Eiken Toxoreagent latex agglutination test- is a
simpler test
• a quantitative test in microtitration plates
the latex particles are coated with inactivated T. gondii
soluble antigen.
A positive control is included in each test kit.
o Interpretation of test results
Interpretation is difficult because of the high
prevalence of antibodies in most populations
due to past and subclinical infections
detection of Toxoplasma specific IgM is
indicative of recent infection
In diagnosing congenital toxoplasmosis
• + of IgM (which does not cross the placenta)
in the infant’s circulation is diagnostic
• Specific IgG in the infant’s circulation may be
of maternal origin or due to infection.
Testing of the infant’s blood at 2 monthly
intervals will show whether the IgG antibody
level is decreasing
At 6–10 months, the infant’s circulation should
not contain maternal Ig G
persistence of Ig G beyond this time is
indicative of infection in the infant.
Serology……
unreliable in immunodeficient (AIDS) pts
normally IgM and IgG rise simultaneously
IgG - persists for years
IgM - undetectable after “cure”
elevated IgM titer is diagnostic of recent infection
A negative IgG or IgM test excludes Diagnosis
both should be + if acutely infected
If IgG screening is + then do IgM test
a + IgM test confirms acute toxoplasmosis
or current Toxoplasma infection (measure
IgM antibodies, have low specificity and the
reported results are frequently
misinterpreted.
In addition, IgM antibodies can persist for
months to more than one year.
Submit IgG positive sera to reference
aboratory for IgM testing for diagnosis of
acute Toxo
Treatment
pyrimethamine + sulfadiazine
Spiramycin-Pregnant
Prevention:
wear gloves when handling
soil
hand washing after outdoor
activities
cook all meats thoroughly
keep cats
