Protozoology

Parasitology Dept.

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Protozoa
General

• Single celled parasitic animal

• Infects predominantly humans and other
primates, but mostly free living
• Varies in shape, size, motile organs, and
reproduction
• Protoplasm contributes the living function
o/t cell, consists of cytoplasm and
nucleoplasm
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Protozoa
General

• Due to motile organs, protozoa is classified

into:
1. Amoeba (pseudopodia)
2. Flagellates
3. Ciliates
4. Myonema
• Due to the way of reproduction, it is divided
into:
1. Sexual
2. Asexual
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Amoebae

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Amoebae
Classification

• Pathogenic amoebae: • Non pathogenic

1. Entamoeba histolytica amoebae:
2. Entamoeba gingivalis 1. Entamoeba coli
2. Entamoeba hartmani
• Pathogenic free-living 3. Endolimax nana
amoebae: 4. Iodamoeba butschlii
1. Naegleria gruberi 5. Dientamoeba fragilis
2. Achantamoeba
castellani
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Entamoeba histolytica
Terminology

• Superclass: Sarcodina
• Class : Rhizopodea
• Order : Amoebida
• Family : Entamoebidae

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Entamoeba histolytica
General

• World-wide distribution, especially in the

poor hygiene areas
• Potentially the most pathogenic intestinal
protozoa
• Causes amebiasis (or amoebiasis),
amoebic dysentery

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Entamoeba histolytica
Morphology

• E. histolytica has two stadiums

during the life cycle:
1. Trophozoite (involves histolytic and
minuta form)
2. Cyst
• Motile apparatus is pseudopodium
(ectoplasm)

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Entamoeba histolytica
Morphology

Trophozoite
• Found in the large intestine and cecum
• The feeding stage o/t parasite
• Involves histolytic (metacystic) and minuta
(precystic) form
Cyst
• Free living resistant stage
• Infective form o/t parasite
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 Entamoeba histolytica
Morphology

Histolytic trophozoite is 20-

40m:
one entamoeba nucleus
with central karyosome
(green arrow)
large pseudopodium (blue
arrow) as the motile organ
endoplasm contains
erythrocytes and granules

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Entamoeba histolytica
Morphology

Trophozoite with ingested RBC

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E. histolytica
Morphology

Trophozoite in
vary feature and
stain
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Entamoeba histolytica
Morphology

Precyst (minuta trophozoite)

Sized 10-20 m
Less motile and rounding
up
No erythrocyte is found in
the endoplasm, only
rough granule, glycogen
vacuole, and chromatoid
bars present (arrowed)

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Entamoeba histolytica
Morphology

Cyst
Sized 5-20 m
Four daughter nuclei
(arrowed)
Chromatoid bars are
occasionally present
Thin wall
Remain viable for about 10
days and up to 30 days in
water
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E. histolytica
Life Cycle

Relatively simple
and direct being
transmitted from
one host to the next
via a free-living
resistant stage -
the cyst 15
Amoebiasis Coli

• Caused by Entamoeba histolytica

• Humans are infected by ingesting cysts, most
often via food or water contaminated with human
fecal material
• Trophozoites live in the host large intestine
• Cysts survive outside the host in water and soils
and on foods
• E. histolytica is potentially the most pathogenic
protozoa in human intestine
• Clinical incubation period 1-14 wks
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Amoebiasis Coli
Epidemiology

• Widely distributed,mostly in poor hygiene areas

• High frequency in high population places
• Carrier persons play role as main source of infection
• Flies, cockroaches, and carrier food-handlers are
considered to be vectors
• Contaminated water is known as mediator of
infection
• It is also frequently diagnosed among homosexual
men
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Entamoeba histolytica
Epidemiology

• It is known that the cysts can pass through the

intestines of flies and cockroaches unharmed
and therefore, there is the possibility that
these insects may act as paratenic hosts
• The association of these insects with the
domestic environment, and in particular their
attraction to food, could mean they play a role
in dissemination of the infective stages of
Entamoeba
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Amoebiasis Coli
Nature of Disease

• Infection may be accompanied by:

1. No symptoms
2. Vague gastrointestinal distress
3. Dysentery (w/ blood and mucus)
• Complications include:
1. Ulcerative and abscess pain
2. Intestinal blockage (rare)
3. Trophozoites infect other organs (mostly liver) 
amoebic liver abscess
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Amoebiasis Coli
Pathology

• Intestinal invasion: ulcerous flask shaped

lesions, may lead to perforation
• Extra intestinal amoebiasis: various
organs, mostly liver (typical abscess)
• Colon ulceration can spread to other sites
by such ways:
1. Percontinuitatum (to secondary sites)
2. Hematogen (to distant organs)

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Amoebiasis Coli
Diagnosis

• Based on clinical symptoms

• Beware of carrier person!
• Stool examination: normal saline, lugol,
trichrom, and hematoxiline stains, or
concentration method
• Serologis test, especially for extra intestinal
amoebiasis patients
• The parasite must be distinguished from other
parasitic (but non-pathogenic) protozoa

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Amoebiasis Coli
Treatment

• Drug of choice: metronidazole

•800mg tid for 10 days
•A combined treatment with the antibiotic tetracycline
and di-iodohydroxyquinine has been successfully
used
• Emetin bismuth iodide
• Carbason
• Milibis
• Niridazole
• Chloroquin
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Amoebiasis Coli
Preventive Treatment

• Treat the patients and carriers

• Health education
• Proper meal preparation and storage
• The use of proper toilets, drinking water
facilities, and garbage cans

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Entamoeba gingivalis
General

• Firstly found in 1849, but became more

important since World War I
• Present in the plaque of patients with
destructive gum disease
• The trophozoite is also found in teeth and
sometimes tonsils
• No cysts are formed and transmission is
entirely by oral-oral contact
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Entamoeba gingivalis
Morphology

• Morphologically
similar to Entamoeba
histolytica
• About 3-10x larger
than a white blood
cell (PMN)
• It attacks and
destroys living red
and white blood
cells
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Entamoeba gingivalis
Morphology

PMN leokocytes (blue arrows) surrounding

the parasite
PMN shown in the endoplasm (red arrows)
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Entamoeba coli
General

• Non pathogenic amoeba, it is commensal

of the human large intestine
• Looks much like E. histolytica, thus it is
important that the two species be
differentiated
• The trophozoite is often found in the feces
of dysenteric patients

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Entamoeba coli
Morphology

Trophozoite
• Sized 15-30m
• Small pseudopodium
• No erythrocytes in the
endoplasm
• Single nucleus with eccentric
karyosome (green arrow) and
irregular chromatin
• Similar to E. histolytica
precyst
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Entamoeba coli
Morphology

Cyst
• Sized 12-20m
• Up to 8 nuclei,
regarding to the
development
• Thick wall

Life cycle is similar to

that of E. histolytica
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Non-Pathogenic Amoeba
Entamoeba hartmanni

• Very similar to
Entamoeba
histolytica, only
much smaller
• 6-10 m in size

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Non-Pathogenic Amoeba
Endolimax nana

• This is a very small

amoeba (6-15um) • Mature cysts contain 4
nuclei
• It has a large, eccentric
endosome and thin
nuclear envelope

Trophozoite
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Non-Pathogenic Amoeba
Iodamoeba butschlii

• Considered to be a commensal
protozoa
• It is found in pigs as well as humans
• The infection can be transmitted from
pigs to humans

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Non-Pathogenic Amoeba
Iodamoeba butschlii

Trophozoite
• Measures usually 12 to 15
µm (range 8 to 20 µm)
• Has one nucleus with a
large, usually central
karyosome
• Cytoplasm is coarsely
granular, vacuolated and
can contain bacteria,
yeasts or other materials

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Non-Pathogenic Amoeba
Iodamoeba butschlii

Cyst
• Measures usually 10 to 12
µm (range 5 to 20 µm)
• Shape varies from ovoidal
to rounded
• Has only one nucleus with a
large, usually eccentric
karyosome
• Characteristically contain a
large glycogen vacuole
(iodophil body) arrowed
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Non-Pathogenic Amoeba
Dientamoeba fragilis

• Live in the large intestine of humans

• No cyst stage has been reported
• There is evidence that this organism is,
in fact, transmitted among humans in
the eggs of human pinworms
(Enterobius vermicularis)

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Non-Pathogenic Amoeba
Dientamoeba fragilis

• Key features used to

differentiate this species
is the presence of two
nuclei (di-entamoeba)
• The smallest parasite of
human gastrointestinal
tract, measures only 3-10
microns

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Non-Pathogenic Amoeba
Dientamoeba fragilis

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Ciliatae

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Ciliates
Classification

• Mostly free-living
• Only one acts as pathogenic parasite
in man: Balantidium coli
• Motile organ is ciliae

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Balantidium coli
General

• Parasitizes in man, pig, rodent, and many

other animals
• Can be transmitted readily among those
species
• Found in host large intestine and colon
• The biggest protozoa in human intestine
• Contains trophozoite and cyst stage

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Balantidium coli
Morphology

Trophozoite
• Oval or bean-shaped
• Measures 60-150
microns in length
• Covered by ciliae
• Contains cytostome,
bean-shaped
macronucleus,
micronucleus, and
contractile vacuoles

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Balantidium coli
Morphology

Cyst
• Rounded with thick wall
• Measures 40-70 m
• Ciliae still exist inside
the wall
• Contains macronucleus,
micronucleus, and
vacuoles

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B. coli
Life Cycle

• Direct life cycle

• Balantidiasis is
often occurs in
swine, and rare
in human
• Swine is
considered as a
potential
reservoir host
for human
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Balantidiasis Coli

• Caused by Balantidium coli, a ciliate

protozoa
• The parasite live in large intestine
• Occurs in humans, swine, rodents, and in
many other mammals, and can be
transmitted readily among these species

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Balantidiasis Coli
Nature of Disease

• It can give various features, from asymptomatic,

mild to severe, according to the virulence o/t
parasite
• Mild illness include gastrointestinal discomfort or
mild diarrhea
• Trophozoites attack intestinal epithelial tissue,
causing flask-shaped ulcers (similar to amebic
ulcers) and secondary bacterial infection
• Ectopic infection can also occur

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Balantidiasis Coli
Diagnosis

• Clinical features
• Stool examination
• Biopsy occasionally needed in ulcer
cases or ectopic infections

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Balantidiasis Coli
Treatment

• Drug of choice: metronidazole

• Preventive treatment is far more
important to prevent the disease

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Flagellatae

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Flagellates
Classification

• Intestinal flagellates • Reproductive tract

 Pathogenic flagellates
flagellates:
Trichomonas vaginalis
Giardia lamblia
 Non-pathogenic
flagelates: • Blood flagellates
1. Chilomastix mesnili
2. Trichomonas hominis 1. Trypanosoma spp.
3. Retortamonas 2. Leishmania spp.
intestinalis
4. Enteromonas hominis

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Flagellates
Classification

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Giardia lamblia
Terminology

• Superclass: Mastigophora
• Class : Zoomastigophorea
• Family : Hexamitidae

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Giardia lamblia
General

• In Europe, it is sometimes referred to as

Lamblia intestinalis
• Consists of two stadiums (trophozoite and
cyst) during the life cycle
• Trophozoites live in the small intestine of the
host
• A flagellate – intestinal flagellate – which
moves with the aid of flagellae

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Giardia lamblia
Morphology

Trophozoite
• Average about 15 µm in
length
• Has a distinct "tear-drop"
shape
• Bi-nucleated, two suckers
in the concave anterior
part, two parabasal bodies,
two axostyle, and 4 pairs of
flagellas

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Giardia lamblia
Morphology

Cyst
• Average 8-13 µm in size
• Ovoid shape
• Immature cyst contains 2
nuclei, and the mature 4
nuclei, and remnants of
the axostyle
• Thick and transparent wall

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Giardia lamblia
Morphology

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Giardia lamblia
Life Cycle

• Cyst are taken in

orally, usually via
contaminated
drinking water
• The trophozoites
may divide by
binary fission
• Trophozoites can
encyst and pass
out with the feces

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Giardiasis Intestinalis

• Caused by Giardia lamblia, a flagellate

• Regarded as the most common flagellate
in the human digestive tract and is highly
contagious
• Found throughout the world
• Traveler’s diarrhea
• Trophozoites live in the small intestine of
the host
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Giardiasis Intestinalis
Nature of Disease

• G. lamblia does not invade the tissues of

the small intestine
• Patients harboring this protozoan can be
asymptomatic carriers
• Or exhibit all or some of the following
symptoms: diarrhea, dehydration,
abdominal pain and weight loss

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Giardiasis Intestinalis
Nature of Disease

• Type of diarrhea:
1. no blood loss
2. fatty consistency as a result of fat malabsorption
3. occurs in heavy infections where attached
trophozoites can cover much of the intestinal
epithelial surface
• The symptoms may be associated w/ the strain
of variable virulence and host immune response

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Giardiasis Intestinalis
Diagnosis

• Stool examination: visualizing both

troph. or cyst in stained/unstained
preparation
• Concentrated method: floatation or
sedimentation
• ELISA

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Giardiasis Intestinalis
Treatment

• Drug of choice: metronidazole

• Quinacrin hydrochloride
• Furazolidone
• Albendazole

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Giardiasis Intestinalis
Preventive Treatment

Similar to those w/ other intestinal protozoa:

• The most important is to avoid using
contaminated drinking water or food
• Treat the patients and carrier persons
properly
• Health education

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Non-Pathogenic Flegellate
Chilomastix mesnili

• Cosmopolitan, but most frequently in warm

climates
• Considered non-pathogenic although the
trophozoite has been associated with
diarrheic stool
• Live in colon and large intestine o/t host
• Direct orofecal route of infection, man is
infected by ingesting cyst
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Non-Pathogenic Flegellate
Chilomastix mesnili

Trophozoite
• The largest flagellate found in man
• Pear shaped, measures 6-20m in length
• Contains one large nucleus, cytostome, axostyle,
flagellum, and undulating membrane

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Non-Pathogenic Flegellate
Chilomastix mesnili

Cyst
• Lemon shaped, measures
6-9m
• Prominent side knob
• Contains single nucleus,
cytostome, and undulating
membrane
• Very resistant, survives in
months in the water

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Non-Pathogenic Flegellate
Trichomonas hominis

• Cosmopolitan
• Considered to be non-pathogenic, although
it has been associated with diarrheic stools
• Found in a wide host range including human
and non-human primates, cats, dogs and
various rodents
• Live in colon o/t host
• Forms no cystic stage
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Non-Pathogenic Flegellate
Trichomonas hominis

• Pyriform shaped, measures 5-15m in

length
• Contains 5 anterior flagella, axostyle, and
undulating membrane which extends the
entire length of the body and projects like a
posterior free flagellum
• Single nucleated

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Non-Pathogenic Flegellate
Retortamonas intestinalis

• Cosmopolitan
distributed, but rarely
encountered
• Live in colon o/t host
• Has trophozoite and
cystic stage
• Pear-shaped
trophozoite, 2 flagellae

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Non-Pathogenic Flegellate
Enteromonas hominis

• Cosmopolitan distributed, but rarely

encountered
• Live in colon o/t host
• Has trophozoite and cystic stage
• Oval trophozoite, 3 anterior and 1 posterior
flagella

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Trichomonas vaginalis
General

• Parasitizes in genitourinary tract in both

males and females
• It may represent from 10-50% of women
presenting with vaginal symptoms
• World-wide distribution
• Sexually transmitted disease

70
Trichomonas vaginalis
Morphology

• It requires no cystic stage

• Trophozoite is pyriform-
shaped and single
nucleated
• Has 4 anterior flagella,
cytostome, axostyle,
undulating membrane, and
1 posterior flagellum

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 T. vaginalis
Life Cycle

• Transexually
infection
• Occasionally
occurs via
indirect
transmission,
i.e. through
contaminated
fabrices
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Trypanosoma spp.
Hemoflagellates

• Family Trypanosomidae
• 3 species known to be pathogenic in
human are:
1. Trypanosoma gambiense
2. Trypanosoma rhodesiense
3. Trypanosoma cruzi
• Causes human sleeping sickness

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Trypanosoma gambiense
General

• Causes Mid African trypanosomiasis

• Vector: Tse Tse fly (Glossina palpalis)
• Final host: humans
• Reservoir host: pig, goat, and cattle
• The parasite lives in blood plasma, lymph nodes,
spleen and brain
• Trypanosoma can also be transmitted through
blood transfusions, organ transplantation,
transplacentally, and in laboratory accidents
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Trypanosoma gambiense
Morphology

• Fusiform shaped
• Flagellated

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Trypanosoma gambiense
Morphology

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Trypanosoma gambiense
Life Cycle

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Trypanosoma rhodesiense
General

• Causes East African trypanosomiasis

• Vector: Tse Tse fly (Glossina morsitans)
• Final host: humans
• Reservoir host: antelopes (wild animal)
• Morphologically similar to the
proceeding species

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 Trypanosoma cruzi
General
• Causes South American trypanosomiasis or
Chagas disease
• Vector: Triatomine bug
• Final host: humans
• Reservoir host: pets, rodents, monkeys,
armadillos
• Living in two forms in human body:
1. trypanosoma form (trypomastigote) found in
peripheral blood vessels
2. leishmania form (amastygote) found in muscles,
brain, reticuloendothelial system, and lymph
nodes 79
Trypanosoma cruzi
Life Cycle

80
Trypanosoma cruzi
Life Cycle

Trypomastigote injected Trypomastigotes

to the next human penetrate human
various cells

Epimastigote Transforms into

amastigote within the
cells

Turn to be
Trypomastigote ingested trypomastigote in the
by triatomine bug blood stream
81
Leishmania spp.
General

• Class Zoomastigophorea
• Order Kinetoplastida
• Family Trypanosomidae
• 3 species known to be pathogenic in
humans are:
1. Leishmania donovani
2. Leishmania tropica
3. Leishmania braziliensis
82
Leishmania donovani
General

• Causes leishmaniasis visceral, Kala-azar

disease, black water fever
• Final host: humans
• Habitat is human’s reticuloendothelial
cells
• Vector: Phlebotomus fly (sandfly)
• Reservoir host: canine
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Leishmania donovani
Morphology

• During the life cycle, Leishmania has two

stages:
1. Leishmania stage (amastigote) found in
humans’ and canine’s RE cells
2. Leptomonas (promastigote) stage found in
vector’s intestine
• Amstigote is rounded or ovoid, measured
2-4 microns, contains kinetoplast,
blepharoplast, and rizoplast
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Leishmania donovani
Morphology

Promastigotes
85
Leishmania donovani
Life Cycle

86
Leishmania donovani
Life Cycle

Promastigote injected Promastigotes

to the next human phagositized by
macrophages

Transform into Transforms into

promastigotes amastigote within the
macrophages

Amastigotes ingested by Amastigotes multiply in

sandfly various cells
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