ACTIVITY 1

Intestinal Protozoa
Objectives:
• To recognize the trophozoite and cyst stages of different
intestinal protozoans.
• To differentiate closely related species based on the
organism’s distinctive characteristics or diagnostic feature.
• To develop practical knowledge on preventive measures
against infections by learning about the life cycle of
intestinal protozoans.
Intestinal Protozoans in Humans
• Entamoeba histolytica
• Entamoeba coli
• Iodamoeba butschlii
• Endolimax nana
• Giardia lamblia
• Chilomastix mesnili
• Balantidium coli
• Cyclospora cayetanensis
• Cryptoporidium parvum
• Isospora belli
TROPHOZOITE CYST
• The motile, feeding stage • Should drying occur, many
• Stage where protozoa parasites have the ability
move by use of flagella, to ENCYST.
cilia, or pseudopodia
(roughly used as basis of Encystment
classification). • A process whereby trophozoite
condenses into a sphere.
• Pre-cyst secrete a resistant
cyst wall around themselves to
survive harsh conditions away
from a host.
• nuclear division begins after
encystment
 Entamoeba histolytica
Entamoeba dispar
Entamoeba coli
Entamoeba hartmanni
Entamoeba polecki
Entamoeba gingivalis (oral)
Endolimax nana
Iodamoeba bütschlii
Entamoeba histolytica and E.dispar
2 morphologically similar amoebas in man.
E. histolytica
• the pathogenic or invasive form
• invades the intestinal mucosa and produces dysentery
or amoebomas
• may give rise to extra-intestinal lesions via the blood,
mainly to the liver.

E.dispar
• The commensal or non-invasive luminal form where
the parasite causes no signs or symptoms of disease.
Entamoeba histolytica
AMEBIASIS (amebic dysentery, amebic hepatitis)
TROPHOZOITE - 20 to 30 µm in
diameter.
• cytoplasm consists of clear
ectoplasm, finely granular
endoplasm;
• food vacuoles often containing
rbc's are common
• characteristic structure is the single
nucleus with one endosome.
Entamoeba histolytica trophozoites
Entamoeba histolytica
CYST - 10-20 µm
Usual range, 12-15 µm.

• Usually spherical.

• Nuclei: 4 in mature cyst. Immature

cysts with 1 or 2 occasionally seen.

• Peripheral chromatin: fine,

uniform granules, evenly
distributed.

• Endosome: Small, discrete, usually

centrally located.

• Chromatoid bodies: Elongated bars

with bluntly rounded ends.
Entamoeba histolytica cysts

uninucleate cyst binucleate cyst

 Entamoeba histolytica life cycle
Infection occurs when infective
cysts are ingested in food or
water that has been
contaminated with human feces.
Excystment:
Metacystic ameba undergoes
multiple fission to form 8
trophozoites.
Trophozoites move into the large
intestine and invade the mucosa.
Encystment:
Trophozoite condenses into a
sphere.
Cysts remain viable in moist
environment for 1 month.
 Entamoeba histolytica pathology
1. COLONIZATION OF THE
LARGE INTESTINE

Flask-shaped lesions: due to

enzymatic degradation of
tissue.

Proteolytic enzymes

Ulcers may form sinuses and

extend into the submucosa
Non-pathogenic Amebae
Entamoeba coli
• Often co-exists with E. histolytica
but is a commensal.

TROPHOZOITE - 20 to 30 m in
diameter
• granular endoplasm is coarser than
E. histolytica
• Nucleus: Endosome is off-centered
or eccentric; with coarse peripheral
chromatin
• Lives in large intestine and feeds
on bacteria and any other cells
available to it; does not invade
tissue.
Entamoeba coli
CYST - encystment is similar to that
of E. histolytica
• Immature cysts are rare in fecal
smears, has 2 nuclei
• Mature cyst is large, 10 to 33 m,
has 8-16 nuclei
• Chromatoidal bodies, if present,
have splinter-like ends (disappear
in most cysts)
• Cyst is released in the feces into
the external environment
Entamoeba coli life cycle
Other Entamoeba species:
Commensals/non-pathogenic forms
E. dispar – identical with E.histolytica but non-invasive.

E. hartmanni - “small-race” E.histolytica; 5-10 m

Usual range, 6-8 m; four nuclei and chromatoidal bars with
smooth, rounded ends
E. gingivalis - harmless commensal, very similar
morphologically to E.histolytica, that lives in many mouths. It
forms no cysts. Opportunistically, it will ingest sloughed host
epithelial cells, bacteria, leukocytes and erythrocytes

E. polecki - infects pigs and monkeys; occasionally in humans

Endolimax nana
TROPHOZOITE - small in size; 6 - 15
µm (usually under 10 m).
• Nucleus: has large blot-like
karyosome, and the lack of
peripheral chromatin.
• Moves slowly; feeds on bacteria and
food debris.
CYST - forms as feces dehydrates
• Small in size (5-10 mm.
Usual range, 6-8 mm.); Spherical,
ovoidal, or ellipsoidal.
• Contains four nuclei that are much
smaller than the nuclei of the
trophozoites and do not have
peripheral chromatin.
Endolimax nana life cycle
Iodamoeba butschlii
• World wide distribution although
not as common as E. coli or E.
nana.
TROPHOZOITE - 9 to 20 m in
diameter.
• actively motile.
• nucleus with a large karyosome is
evident.
• Chromatin bodies form striations
around the karyosome.
• The cytoplasm appears granular
containing vacuoles with ingested
bacteria and debris.
Iodamoeba butschlii
CYST
• 5-20 m. Usual range, 10-12
m
• one nucleus in mature cysts
usually eccentrically placed.
• Chromatoid bodies are not
present.
• Glycogen is present as a
compact well defined mass
staining dark brown with
iodine.
Iodamoeba bütschlii life cycle
 Harmless:
Accurate identification of these
parasites are important:
Endolimax nana indicators of fecal-oral
contamination, and
could be confused with the
pathogenic species;
• leading to superfluous
Iodamoeba butschlii
treatment for a non-pathogenic
parasite
or
• omission of appropriate
therapy for a pathogen.
Entamoeba coli
 Endosome
or
Karyosome

Endolimax nana
Entamoeba histolytica

Endosome
or
Karyosome

Iodamoeba butchlii

Entamoeba coli
 Giardia lamblia
Dientamoeba fragilis
Chilomastix mesnili
Enteromonas hominis
Retortamonas intestinalis
Trichomonas hominis
Giardia lamblia / G. intestinalis
GIARDIASIS
• the most common identified cause of
water-borne disease associated with
▫ breakdown of water purification systems
▫ drinking from contaminated streams
▫ travel to endemic areas (Russia, India, Rocky
Mountains, etc.)
▫ day care centers.
Giardia lamblia / G. intestinalis
TROPHOZOITES
• Size: 10-20 m.
Usual range, 12-15 m.
• Half pear-shaped organism
• 8 flagella and 2 axostyles
arranged in a bilateral
symmetry
• There are two anteriorly
located large suction discs
• Cytoplasm contains two nuclei
and two parabasal bodies
• Endosome: central
• Motility: “Falling leaf.”
Morphology of Giardia lamblia trophozoite

Light microscope photos of trophozoites

Giardia lamblia / G. intestinalis
CYST
• Size: 8-19 m.
Usual range. 11-12 m.
• Shape: Oval or ellipsoidal.
• Nuclei: Usually 4; Usually located
at one end.
• Endosome: centrally located
• Deep staining fibers or fibrils may
be seen lying laterally or obliquely
across fibrils in lower part of cyst.
• Thin cyst wall
Giardia lamblia cyst
Giardia lamblia life cycle
 Pathology of Giardia lamblia infections

1.Abdominal pain accompanied by nausea,

flatulence, and weight loss occurs in most cases.
•Diarrhea caused by the production of a watery
mucus in response to the trophozoites (foul-
smelling bulky, explosive, often watery).
2. Numerous trophozoites cover and shorten
microvilli of intestinal cells causing malabsorption of
nutrients.
Chilomastix mesnili
- nonpathogenic/ commensal flagellate
TROPHOZOITES - 6-24 m.
Usual range, 10-15 m.
• Pear shaped.
• 1 nucleus; not visible in
unstained mounts.
• Flagella: 3 anterior and 1 in
cytosome.
• Motility: Stiff, rotary.
• Prominent cytostome
extending 1/3-1/2 length of
body. Spiral groove across
ventral surface.
Chilomastix mesnili
CYST - 6-10 m.
Usual range, 8-9 m
• Lemon shaped with anterior
hyaline knob.
• 1 nucleus; Not visible in
unstained preparations.
• Cytostome with supporting
fibrils. Usually visible in
stained preparations.
• common diagnostic stage in
feces.
 Chilomastix mesnili life cycle
1. Trophozoites live in the human large
intestine and multipy by fission

2. Feces dehydration in the large intestine

stimulates encystment.
Cyst is released in the feces.

3. Cyst survives in water in the external

environment.

4. When cyst is ingested, it excysts in the

small intestine and moves to the large
intestine.

Transmission involves ingestion of cysts

primarily from food and water
contaminated with feces.
Balantidium coli
the only parasitic ciliate in man.
TROPHOZOITE - 50-70 m or
more. Usual range, 40-50 m.
• Ovoid with tapering anterior
end.
• Nuclei : 1 large, kidney shaped
macronucleus and 1 small
micronucleus immediately
adjacent to macronucleus.
• Body surface covered by spiral,
longitudinal rows of cilia.
• Contractile vacuoles are present.
• Motility: Rotary, boring.
Oral ciliature

Somatic ciliature

Balantidium coli
Trophozoite
Balantidium coli
CYST - 45-65 m. Usual range,
50-55 m.
• Spherical or oval.
• 1 large macronucleus visible in
unstained preparations as hyaline
mass.
• Macronucleus and contractile
vacuole are visible in young
cysts. In older cysts, internal
structure appears granular.
 The Coccidia (Apicomplexa)
Coccidians are members of the Phylum Apicomplexa that
infect the intestinal mucosa (not rbc’s) of the vertebrate
host.

Cryptosporidium spp.
Cyclospora cayetanensis
Isospora belli
Sarcocystis hominis
Life Cycle of Cryptosporidium
Cryptosporidium spp.
• Oocysts are the infective and diagnostic stage – size is 3-6 µm
Usual range, 4-5 µm in diameter .
• Sporulated oocysts, containing 4 sporozoites, are excreted by the infected
host through feces and possibly other routes such as respiratory
secretions
• The sporozoites are released and parasitize epithelial cells of the
intestines. Here, parasites undergo asexual multiplication (schizogony or
merogony).
• Sexual multiplication (gametogony) follows, producing microgamonts
(male) and macrogamonts (female).
• Fertilization results into oocysts which sporulate in the intestine.
• Spherical or oval oocyst are shed by the host and is the infective stage.
Cryptosporidium parvum

Oocysts on the
surface of small
intestinal cells
 Oocysts of Cryptosporidium

Each oocyst contains 4 sporozoites that attach to surface of small intestinal cells

unstained acid red stain

Oocysts from cattle feces – oocysts are 4-5 µm in diameter
 Isospora belli

LIFE CYCLE is similar to Cryptosporidium:

• Trophozoites, merozoites, & gametocytes

occur in the cytoplasm of small intestinal cells
(not on surface like Cryptosporidium)

• Oocysts (20 -30 µm long) are released

in feces (diagnostic)

• Oocysts are infective to new human

host via human fecal contamination.
 Organism Transmission Symptoms Diagnosis Treatment

Dysentery with blood Stool: cysts with 1-4 GI: Iodoquinol or

Entameba
Oro-fecal and necrotic tissue. nuclei and/or trophs. Metronidazole
histolytica
Chronic: abscesses Trophs in aspirate. Abscess: Metronidazole

Fowl-smelling, bulky Stool: typical old man

Iodoquinol or
Giardia lamblia Oro-fecal diarrhea; blood or giardia troph and/or
Metronidazole.
necrotic tissue rare. cyst.

Dysentery with blood

Oro-fecal; Stool: ciliated trophs Iodoquinol or
Balantidium coli and necrotic tissue but
zoonotic and/or cysts. Metronidazole.
no abscesses.

Cryptosporidium Paromycin
Oro-fecal Diarrhea Ooocysts in stool
parvum (investigational)
Isospora belli Oro-fecal Giardiasis-like Ooocysts in stool Sulpha drugs

Trichomonas Vaginitis; occasional Flagellate in vaginal (or Mebendazole; vingar

Sexual
vaginalis urethritis/prostatitis. urethral) smear. douche; steroids
Fecal-oral transmission of the pathogens

• the most common mode of GI infections

• water, food and hands become contaminated with
fecal material which then come in contact with the
mouth.
Fecal-Oral Transmission Factors
• poor personal hygiene
▫ children (eg, day-care centers)
▫ institutions (eg, prisons, mental hospitals, orphanages)
▫ food handlers
• developing countries poor sanitation
▫ lack of indoor plumbing
▫ endemic
▫ travelers' diarrhea
• water-borne epidemics
▫ water treatment failures
• male homosexuality
▫ oral-anal contact