Amoeba

AMOEBA
AMOEBA
• From the Greek word amoibē meaning “change”
• Singular: Amoeba or less commonly – ameba
• Plural: Amoebae or amebae
• Is a type of cell or unicellular organism which has the
ability to alter its shape, primarily by extending and
retracting pseudopods
• Commonly found in water and soil, possessing no set cell
organs, structure or defining shape
Amoeba
Non-
pathogenic Pathogenic
Intestinal Mouth Intestinal Free-living
Entamoeba Entamoeba
coli dispar Entamoeba Entamoeba Acanthamoeba Naegleria
gingivalis spp. spp.
histolytica
Entamoeba Entamoeba
hartmanni polecki
Iodamoeba Entamoeba Entamoeba Endolimax

buetschlii moshkovskii chattoni nana
CLASSIFICATION
Kingdom: Protozoa
Subkingdom: Protozoa
Phylum: Sarcomastigophora
Subphylum: Sarcodina
Superclass: Rhizopoda
Class: Lobosea
Order: Amoebida
Family: Entamoebidae
Genus: Entamoeba
Endolimax
Iodamoeba
MAJOR STAGES IN LIFE CYCLE
1. Cyst
2. Trophozoite
BASIC STRUCTURES
BASIC STRUCTURES
Genus Entamoeba
• Spherical nucleus with a comparatively small karyosome
near or at the center of the nucleus
• Peripheral chromatin granules line the distinct nuclear
membrane
• Delicate achromatic threads (fibrils) connect the
karyosome to the nuclear membrane
Genus Endolimax
• Vesicular nucleus with a relatively large karyosome of
irregular shape
• Several achromatic threads (fibrils) connects the
karyosome to a delicate nuclear membrane
• Thin layer of peripheral chromatin which is
inconspicuous
Genus Iodamoeba
• Large, chromatin–rich karyosome surrounded by a single
layer of periendosomal granules (achromatic globules)
and anchored to the nuclear membrane by radiating
achromatic threads (fibrils)
PATHOGENIC
AMOEBAE
INTESTINAL AMOEBAE
Entamoeba histolytica
“Tissue Invading Amoeba”
Synonyms: Entamoeba dysenteriae, Amoeba dysenteriae,
Entamoeba tetragena, Endamoeba histolytica,
Endamoeba dysenteriae
• Pathogenic amoeba of man causing amoebiasis, amoebic
dysentery, amoebic hepatitis or liver abscess, amoebic
brain and lung abscess
Morphology, Biology and Life Cycle

 5 stages (Cyst, Metacyst, Metacystictrophozoite or
Amoebulae, Trophzoite, Precyst)
Trophozoite:
 Invasive, growing, feeding stage
 Size: 12-60 μm
 Exhibits remarkable locomotion (progressive,
directional movement) with pseudopodia as locomotory
organelles
 Cytoplasm is described as having a ground glass
appearance and is differentiated into a clear outer
ectoplasm and inner finely granular endoplasm in which
food vacuoles containing ingested red blood cells may be
observed
Trophozoite:
 Nucleus is spherical and contains
a small distinct dot like central karyosome surrounded by
an unstained halo and anchored by numerous delicate,
radiating achromatic fibrils to the inner surface of the
nuclear membrane (spoke of a wheel/bull’s eye
appearance)
 Lining the nuclear membrane is a rim of chromatin that
is regularly/uniformly arranged in small round granules
Cyst:
 Size: 10-20 μm
 Spherical, may be sub spherical or ovoidal
 Highly refractile cyst wall
 Mature: 4 nuclei with centrally located karyosome and
no cytoplasmic inclusions
 Young: 1-2 nuclei, glycogen mass with hazy margins and
long or short chromatoidal bodies which are rod-shaped
(cigar or sausage-shaped)
Cyst:
Habitat:
 Cecum
Mode of Transmission:
 Ingestion of contaminated food and drinks with feces
containing the infective stage
Contamination may be through:
1. Polluted water supply
2. Unclean handling of infected individuals
3. Droppings of flies and other insects
4. Use of human excreta in vegetable gardens
5. Gross carelessness in personal hygiene in children’s
asylums, mental hospitals, prisons and other places
Pathogenesis and Clinical Manifestations:
Proposed Mechanisms for Virulence Production:

1. Production of enzymes or other cytotoxic substances
2. Contact-dependent cell killing
3. Cytophagocytosis
Pathogenesis and Clinical Manifestations :
 Intestinal Amoebiasis:
E. histolytica trophozoites adhere to the colonic mucosa
through a galatose inhibitable adherence lectin (Gal lectin)
or N-acetyl-d-galactosamine adherence lectin
Then the amoebae kill mucosal cells by the activation of

their caspase-3, leading to heir apoptotic death
engulfment
• Clinically presents as gradual onset of abdominal pain
and diarrhea with or without blood and mucus in stools.
• Fever is not common and it occurs only in one third of
patients.
• Some patients may only have intermittent diarrhea
alternating with constipation.
• Children may develop fulminant colitis with severe
bloody diarrhea, fever, and abdominal pain.
Should be differentiated from bacillary dysentery of the
following etiology:
1. Shigella
2. Sallmonella
3. Campylobacter
4. Yersinia
5. Enteroinvasive Escherichia coli
6. Inflammatory bowel disease
Basis Amoebic Dysentery Bacillary Dysentery
Gross appearance Gelatinous mixture of blood Mucopurulent mass streaked
mucus and feces with blood
Amount Relatively copious Small
Odor Offensive (Fishy) Inoffensive
Color Bright red Dark red
Reaction Acidic Alkaline
Ghost cells None (RBCs are intact) 95% degenerated ghost cells
Macrophages Rarely seen Present
Red Blood Cells Clumped Never clumped, discrete
Charcot-Leyden cystals Present Absent
Bacteria Numerous Nil to none
Pus Cells Scanty Numerous
 Extra-Intestinal Amoebiasis:
• Amoebic liver abscess (ALA) or hepatic amoebiasis is the
most common form
The liver is tender and hepatomegaly is present (50%)
The most serious complications are rupture into the

pericardium, rupture into the pleura, and super infection
• Amoebic liver abscess (ALA) or hepatic amoebiasis
Increased WBC count and liver function test
Aspiration of abscess: punch or needle biopsy
 Characteristic Chocolate colored (anchovy sauce)
content of abscess – mixture of sloughed liver tissue
and blood or degenerated hepatocytes, RBC, leukocytes
• Secondary amoebic meningoencephalitis occurs in 1-2%
of cases, and should be considered in cases of amoebiasis
with abnormal mental status
• Renal involvement caused by extension of ALA or
retroperitoneal colonic perforation is rare
• Genital involvement is caused by fistulae from ALA and
colitis or primary infection through sexual transmission
Diagnosis:
 Standard method: Detection of trophozoites and/or cysts
in stool (NSS/I2)
 Ideally a minimum of three stool specimens (3 or more)
collected on different days at a 3 day or 4 day interval
 Diarrheic/dysenteric/watery stool – Trophozoite
Examined preferably within 30 minutes after passage
 Formed stool – Cyst
Any part of the feces but should include a portion or any
fleck of mucus/blood adherent to the feces
Diagnosis:
 If few cysts are few to be seen, perform concentration
techniques such as Formalin Ether/Ethyl Acetate
Concentration Technique (FECT) or floatation methods like
Zinc Sulfate Floatation Method
Detection of E. histolytica trophozoites with ingested
RBCs is diagnostic of amoebiasis
 Charcot-Leyden crystals can also be seen in stool
Diagnosis:
 Saline-purged specimens:
• Will provide material for (+) diagnosis when routine fecal
examination has been unrewarding
• Sodium sulfate (Glauber salt) or Phosphosoda is preferred
• Following purgation, earlier fecal evacuations are
discarded. Sedimented elements of mucus and tissue cells
from the 2nd or 3rd bowel movements are pipetted onto the
slide, coversliped and examined for trophozoites
Diagnosis:
 Sigmoidoscopy material:
• Maybe expressed from ulcers by means of gentle
pressure from long-handled curette or loop
• 1/3 of lesions are in the sigmoidorectal area
• Scrapings or aspirated from suspected amoebic ulceration
maybe obtained and examined microscopically for motile
trophozoites
Diagnosis:
 Culture:
• Used in the study of metabolism and pathogenicity, and in
the production of antigens for serogdiagnosis
• Culture medium: Dibasic medium of Boeck and Drbohlav
(Egg slant base with isotonic overlay + LES-Locke egg
serum), Diamond’s medium TYI-S-33, Robinson’s and Inoki
medium
 ELISA with PCR: gold standard
Amoebic liver abscess: detection of antibodies is still key
Treatment and Prognosis:
 Two objectives: cure invasive disease and eliminate
passage of cysts
 Metronidazole for invasive amoebiasis (Tinidazole and
Secnidazole are also effective)
 Diloxanide furoate for asymptomatic cyst
passers/carriers; also given after a course of metronidazole
for invasive amoebiasis
Epidemiology:
 True prevalence worldwide is 1-5% of world population
Prevention and Control:
 Improve environmental sanitation:
• Sanitary disposal of human feces
• Secure safe food supply and drinking water
• Proper hygiene
 Vaccines are soon to be available using candidate
molecules:
• Serine-rich E. histolytica protein (SREHP)
• Adherence lectin (Gal/GalNAc lectin)
• 29kDa cysteine-rich amoebic antigen
FREE-LIVING AMOEBAE
CLASSIFICATION
Kingdom: Protozoa
Class: Lobosea
Order: Schizopyrenida
Family: Valkamphidae
Genus: Naegleria
Naegleria spp.
• Free-living thermophilic organisms which thrive best in
hot springs and other warm aquatic environments
• Naegleria fowleri is the causative agent of Primary
Amoebic Encephalitis (PAM)

 Two vegetative forms: An amoeba(trophozoite form),
and a flagellate (swimming form)
 A dormant cyst form is produced when conditions are
not favorable
Naegleria spp.
Vegetative forms:
1. Amoeboid
 Size: 10-35 μm (Rounded: 10-15 μm)
 Nucleus is large and contains a large, dense central
karyosome and lacks peripheral chromatin
 Cytoplasm is granular and contains food vacuoles
 Usually has a single blunt lobopodium and conspicuous
contractile vacuoles
Naegleria spp.
2. Flagellate:
 Elongated shape
karyosome and lacks peripheral chromatin
 Cytoplasm is granular and contains food vacuoles and
conspicuous contractile vacuoles
 Bears 2 long flagella at one end
Naegleria spp.
Cyst:
Spherical shape
 Uninucleated
karyosome
 Cytoplasm is granular
 Penetration of trophozoite to the nasal mucosa
Naegleria spp.
Diagnosis:
 Suspected in persons with a compatible history of
exposure and a rapidly progressive meningoencephalitis
 In the past, definitive diagnosis of PAM was based on
demonstration of characteristic trophozoites in the brain
and cerebrospinal fluid. Aspirates from suspected
infections, when introduced into bacteria-seeded agar
culture medium, will exhibit active trophozoites within
24 hours
 More sensitive and specific for N. fowleri:
PCR and immunostaining
Naegleria spp.
Treatment:
 Amphotericin B in combination with clotrimazole
(synergistic)
 Azithromycin and voriconazole (newer agents
Epidemiology:
 Only one case of PAM has been reported locally
 N. philippinensis which is morphologically
indistinguishable but biochemically distinct from other
known species
Naegleria spp.
Prevention:
 Chlorination of water at 1 ppm or higher
CLASSIFICATION
Kingdom: Protozoa
Class: Lobosea
Order: Amoebida
Family: Hartmannellidae
Genus: Acanthamoeba
Acanthamoeba spp.
• Ubiquitous, free-living amoeba
• Etiologic agent of Acanthamoeba Keratitis (AK) and
Granulomatous Amoebic Encephalitis (GAE)

 Trophozoite stage with characteristic prominent “thorn-
like” appendages (acanthopodia)
 Highly resilient cystic stage
Acanthamoeba spp.
Trophozoite:
 Single large nucleus with a centrally-located, densely
staining nucleolus, large endosome
 Finely granulated cytoplasm and a large contractile
vacuole
 Small spiny filaments for locomotion (acanthopodia)
 Entry of the amebae to the eye, the nasal passages to
the lower respiratory tract, or ulcerated or broken skin
Acanthamoeba spp.
Acanthamoeba spp.
Diagnosis:
Acanthamoeba keratitis
 Epithelial biopsy or corneal scrapings for recoverable
amoeba
 Culture and molecular analysis through PCR (species-
specific: A. castellani, A. culbertsoni, A. hutchetti,
A. polyphaga, and A. Rhysoides)
Acanthamoeba spp.
Diagnosis:
Granulomatous Amoebic Encephalitis
 Usually made post-mortem
 Recovery of amoeba from cerebrospinal fluid is
exceedingly rare, and imaging results are generally
nonspecific
 Specific diagnosis depends on demonstrating
the trophozoites or cysts in tissues using
histopathologic stains and microscopy
Acanthamoeba spp.
Treatment:
Acanthamoeba keratitis
 Historically, only surgical excision of the infected cornea
with subsequent corneal transplantation was curative
 While, early recognition of AK coupled with aggressive
combination anti-amoebic agents can preclude the need
for extensive surgery. D’Aversa and his colleagues have
achieved acceptable results with clotrimazole combined
with pentamidine, isethionate, and neosporin
 Other agents: polymyxin B,ketoconazole, miconazole,
and itraconazole and etc
Acanthamoeba spp.
Treatment:
Granulomatous Amoebic Encephalitis
 Combinations of amphotericin B, pentamidine
isethionate, sulfadiazine, flucytosine, fluconazole or
itraconazole
 Amphotericin, clotrimoxazole, and rifampin
Epidemiology:
 1st case of AK (1990s): a patient from the PGH, and
samples obtained was shown to cause GAE in mice.
Acanthamoeba spp.
Prevention and Control:
 Robust immune system
 Meticulous contact lens hygiene
 Rinsing contact lenses in tap water should be avoided
Morphological Differences
Naegleria / Acanthamoeba Entamoeba histolytica
Nucleolus large and distinct Nucleolus small and indistinct
Contractile vacuoles present Contractile Vacuoles absent
Single nucleus in cystic stage 4 nuclei in mature cyst
No glycogen and chromatoid bodies in Glycogen and chromatoid bodies present
cystic stage in cystic stage
Mitochondria present Mitochondria absent
Cyst wall may have pores or ostioles Cyst wall has no pores
Morphological Differences
Naegleria Acanthamoeba
Trophozoite with broad pseudopods Trophozoite with filamentous pseudopods
Actively motile Sluggishly motile
Forms flagellate stage Does not form flagellate stage
Single-walled cysts Double-walled cysts
Does not encyst in tissue May encyst in tissues
NON-PATHOGENIC /
COMMENSAL
AMOEBAE
Entamoeba dispar
• Morphologically similar to E. histolytica, but their DNA
and ribosomal RNA are different
• Relatively difficult to distinguish from E. histolytica but
can be done through culture, biochemical methods and
polymerase chain reaction (PCR)
• Although considered as non-pathogenic, it is evidently
capable of producing intestinal lesions in experimental
animals
Entamoeba moshkovskii
• First detected in sewage
• Previously known as the “Laredo strain”
• Morphologically indistinguishable from E. histolytica and
E. dispar, but differs from them biochemically and
genetically
• Osmotolerant, able to grow at room temperature (25-
30°C optimum), able to survive temperatures from 0-41°C
• Limited pathogenicity in experimental animals, but is
non-pathogenic to humans
• All human isolates have been found to belong to one
group – “Ribodeme 2”
Entamoeba hartmanni
• Appearance is relatively similar to E. histolytica apart
from its smaller size
• Trophozoite measures 3-12 μm in diameter (E.
Histolytica: 12-60 μm) and does not ingest red blood cells
• Mature cyst measures 4-10 μm and are quadrinucleated,
and have rod-shaped chromatoid material with rounded
or squared ends
Entamoeba polecki
• Found in the intestines of monkey and pigs
• Rarely infect humans, though a high prevalence (19%)
was reported in Papua New Guinea
Trophozoite:
 Motility is sluggish and the karyosome is small and
centrally located
Cyst:
 Consistently uninucleated, and chromatoidal bars are
frequently angular and pointed
Entamoeba chattoni
• Found in apes and monkeys
• Detected in eight human infections
• Morphologically identical to E. polecki
• Identification was done via isoenzyme analysis
Entamoeba gingivalis
“Atrial Amoeba”
Synonyms: Amoeba gingivalis, Amoeba buccalis,
Entamoeba buccalis
Geographic Distribution:
• Cosmopolitan
• 1st parasitic amoeba of man to be described

 Only the trophozoite stage is known
Trophozoite:
 Size: 10-20 μm
 Exhibits a definite zone of demarcation between the
clear ectoplasm and the more granular endoplasm
 Endoplasm has numerous food vacuoles that contain
cellular debris such as partly digested leukocytes,
epithelial cells and bacteria
 Abundant in cases of oral disease
Habitat:
 Lives on the surface of gums and teeth, in gum pockets,
and in tonsillar crypts
 Direct, through kissing, droplet spray, or by sharing
utensils
Diagnosis:
 Demonstration of the trophozoite in material or
scrapings removed from the gingival margin of the gums,
from between teeth or dentures; from the soft tartar of
teeth
Prevention:
 Proper care of teeth and gums
Entamoeba coli
Synonyms:
Amoeba coli, Endamoeba hominis, Loschia coli,
Councilmania lafleuri
• Cosmopolitan
Morphology, Biology and Life Cycle:

 5 stages (Cyst, Metacyst, Metacystictrophozoite,
Trophozoite, Precyst)
Entamoeba coli
Trophozoite:
 Size: 15-50 μm
 Densely granular endoplasm with
numerous food vacuoles, bacteria, and debris having a
“honeycomb” or “dirty-looking” appearance
 Spherical nucleus, with relatively thick nuclear
membrane lined with coarse chromatin granules and a
eccentrically located karyosome
 Movement is typically sluggish and undirected with
board, blunt and short pseudopodia
Entamoeba coli
Cyst:
 Size: 10-35 μm
 Spherical in shape with definite cystic wall
 Mature cyst has 8 nuclei, rarely 16 or 32 with
eccentrically located karyosome and no cytoplasmic
inclusions
 Immature (young) cyst have 1 to 4 nuclei, a glycogen
vacuole, and chromatoidal bars are seen as spicules or
irregular masses of hematoxylin material; broomlike or
rods with splintered ends
Entamoeba coli
Entamoeba coli
Life Cycle:
MATURE CYST -> ingested -> stomach -> small intestine ->
EXCYSTATION -> 8-nucleated mass escapes -> METACYST ->
cytoplasmic division -> 8 metacystictrophozoite -> large
intestine -> feed and grow -> MATURE TROPHOZOITE ->
binary fission -> unfavorable condition -> ENCYSTATION
starts -> undigested food extruded out -> assumes a
spherical shape -> PRECYST -> secrete a tough cystic wall ->
encystation is accomplished -> UNINUCLEATE CYST ->
BINUCLEATE CYST -> QUADRINUCLEATE CYST ->
OCTANUCLEATE CYST -> passed out with feces
Entamoeba coli
Habitat:
 Lumen of the large intestine
 Ingestion of food, drinks or objects contaminated with
the mature cyst
Diagnosis:
 Demonstration of cyst and/or trophozoite in stool
examination
Prevention:
 Personal hygiene and sanitary disposal of human excreta
Entamoeba coli
Endolimax nana
“Dwarf Intestinal Slug”
Synonyms:
Entamoeba nana, Endolimax intestinalis
• Occurs with the same frequency as Entamoeba coli

 5 stages (Cyst, Metacyst, Metacystictrophozoite,
Trophozoite, Precyst)
 Same stages and life cycle as Entamoeba coli except
that mature cysts have only 4 nuclei
Endolimax nana
Trophozoite:
 Size: 5-12 μm
 Delicately granular vacuolated cytoplasm with a narrow
rim of clear ectoplasm
 Projects short, blunt, hyaline pseudopodia
 Food vacuoles are seen to contain bacteria, vegetable
cells, and crystals
 Minute spherical or sub spherical nucleus with
conspicuous irregular karyosome which may be centrally
or eccentrically located
Marginal chromatin is a thin layer
Endolimax nana
Cyst:
 Size: 5-14 μm
 Oval or round
 Immature cyst contains 1 or 2 nuclei, has an ill defined
glycogen mass, and one to few small slightly curved
chromatoidal bodies
 Mature cyst has 4 nuclei, rarely 8
Habitat:
Endolimax nana
Mode of transmission:
 Ingestion of viable mature cysts in polluted water, food,
drinks or from contaminated objects
Diagnosis:
 Stool examination:
Typical ovoidal cysts of E. nana are easily diagnosed
Round cysts and living trophozoites may be difficult to
differentiate from small strains of E. Histolytica
Prevention:
 Personal cleanliness and community sanitation
Endolimax nana
Endolimax nana
Iodamoeba buetschlii
“Iodine Cyst of Wenyon”
Synonyms: Entamoeba wiliamsi, Entamoeba buetschlii,
Endolimax williamsi
• Ordinarily less common than E. coli or E. nana

that mature cysts have typically 1 nuclei (rarely 2)
“Iodine Cyst of Wenyon”
Synonyms: Entamoeba wiliamsi, Entamoeba buetschlii,
Endolimax williamsi
• Ordinarily less common than E. coli or E. nana

that mature cysts have typically 1 nuclei (rarely 2)
Trophozoite:
 Size: 4-20 μm (average: 9-14 μm)
 Short, blunt, hyaline pseudopodia
 Fairly active with progressive movement in freshly
evacuated stool and sluggish in older stool
 Ectoplasm is clear, usually not well differentiated from
the denser endoplasm that contains coarse and fine
granules. Has bacteria, yeast cells, and debris in food
vacuoles
Trophozoite:
 Nucleus is large and vesicular, with a large central and
densely chromatic karyosome, surrounded by achromatic
granules
 Karyosome may appear to have a thin, unstained “halo”
 Nuclear membrane is delicate, devoid of chromatin
granules
Cyst:
 Size: 6-16 μm (about 9-10 μm in diameter)
 Irregularly pyriform or ovoidal
 No increase in nuclei within the cyst, rarely the ripened
cyst may have 2 nuclei
 Most conspicuous feature is a large glycogen vacuole
that is densely packed with glycogen and is distinctly
outlined as an ovoid, polygonal, or broadly reniform mass
that stains golden brown with Iodine (I2)
Habitat:
 Trophozoite feeds on enteric bacteria
 Ingestion of mature cyst
Diagnosis:
 Stool examination
Prevention:
 Better personal hygiene and community sanitation
END
“Remember why you started.
Don’t give up, find a way.”
References: Prepared by:
Medical Parasitology in the Philippines – Belizario, De Leon Justin Zindell S. Fuentes, RMT, ASCP(i)CM
Clinical Parasitology – Faust, Russel, Jung
Parasitology – Noble & Noble
Parasitology, Protozoology & Helminthology – Chatterjee
Textbook of Parasitology – Belding

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AMOEBA

Intestinal Mouth Intestinal Free-living

Iodamoeba Entamoeba Entamoeba Endolimax

Morphology, Biology and Life Cycle

Proposed Mechanisms for Virulence Production:

Then the amoebae kill mucosal cells by the activation of

The most serious complications are rupture into the

Morphology, Biology and Life Cycle

Morphology, Biology and Life Cycle

Morphology, Biology and Life Cycle

Morphology, Biology and Life Cycle:

Morphology, Biology and Life Cycle:

Morphology, Biology and Life Cycle:

Morphology, Biology and Life Cycle:

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