AMOEBA

PROTOZOA
 Form and Function
• Nucleus and cytoplasm
- Plasma membrane; like all cells, the bodies of protozoa are covered
by plasma membrane
- Pellicular microtubules: or fibrils may coarse beneath the plasma
membrane
ex: kinetoplasmid flagellates: microtubules underlie flexible
membrane
ex: trypanosome & trichomonas: adjoining membranes have, a
fibrous connection between them such as between the body and
undulating membrane.
- Protozoa like fungi, plants and animals are describes as eukaryotes;
 Form and Function
• Nucleus and cytoplasm (cont’d)
- eukaryotes: the genetic material – DNA- is carried on well-defined
chromosomes contained within a membrane-bound nucleus
- Mitochondria
ex: amoeba: branched tubular cristae
flagellates: a single, large body
ciliates: arranged as elongated sausage-shaped structures
- Golgi Apparatus (dictysome)
ex: flagellates: large and/or multiple parabasal bodies in
association with kinetosomes, the “basal bodies”
 Form and Function
• Protozoa consist of a single cell
- many species contain more than one nucleus during portions of
their life cycles
 Classification
Protozoa:
- was once a phylum name
- currently used colloquially as a common noun
- refers to a number of phyla
 Form and Function
• Locomotor Organelles
Protozoa move by 3 basic types;
1. Pseudopods (amoeba)
2. Flagella (flagellates)
3. Cilia (ciliates)
* Some amoebas possess both flagella and pseudopods –
transformation from flagellated to amoeboid cell occurs in response to
environmental conditions and is a recognized life-cycle event
 Encystment
Many protozoa can secrete a resistant covering and enter a resting stage
– cyst
Conditions favoring encystment involve some adverse environment
events such as food deficiency, desiccation, increased tonicity, decreased
oxygen concentration or pH or temperature change.
During encystment, a cyst wall is secreted.
During excystation, there is return to a favorable environment.
*in coccidians the cystic form is an oocyst which is formed after gemete
union and in which multiple fission occurs (sporogony) with cytokinesis
to produce sporozoites.
Form and Function
 AMOEBA
I. Entamoeba histolytica
II. Entamoeba hartmanni
III. Entamoeba dispar
IV. Entamoeba coli
V. Entamoeba gingivalis
VI. Entamoeba polecki
VII. Iodamoeba bütschlii
VIII.Endolimax nana
 OTHERS
• Naegleria fowleri
• Acanthamoeba spp (Acanthamoeba castallani)
 Classification
• Protozoa have been divided traditionally on the basis of their means
of locomotion, although this character is no longer believed to
represent genuine relationships:

Flagellates (e.g. Giardia lamblia)

Amoeboids (e.g. Entamoeba histolytica)
Sporozoans (e.g. Plasmodium)
- Apicomplexa
- Microsporidia
Ciliates (e.g. Balantidium coli)
 Feeding and Metabolism
Protozoa lacking chloroplasts are all heterotrophic – get energy from
complex carbohydrates and nitrogen from amino acids
Mouth parts:
amoeba – temporary
ciliates – permanent; cystostome
Excretion: of indigestible material
Ciliates - cytopyge
 Entamoeba histolytica
A. Disease produced
amoebiasis
amebic dysentery
hepatic amoebiasis

B. History
Lambl 1859 – first discovered the parasite
Lösch 1875 – proved its pathogenic nature
Schaudin 1903 – differentiated E. histolytica from E. coli
 Entamoeba histolytica
C. Geographical Distribution
world wide
more common in the tropics and subtropics than in the temperate
zone

D. Habitat
trophozoites live in the mucous and submucous layers of the large
intestines of man
 Entamoeba histolytica
E. Morphology
Several successive stages occur in the life cycle of E. histolytica
1. trophozoite
2. precyst
3. cyst
4. metacyst
5. metacystic trophozoite
 Entamoeba histolytica
E. Morphology
1. Trophozoite
a. size
vary in size: 10-60μm in diameter
average: 20-30μm
b. shape: not fixed – constantly changing
 Entamoeba histolytica
E. Morphology
1. Trophozoite
c. motion
- move by pseudopodia
- progressive and directional active and purposeful
- pseudopodia are cytoplasmic protrusions that may be
formed at any surface of the organism
 Entamoeba histolytica
E. Morphology
1. Trophozoite
d. cytoplasm
- divisible into 2 portions
ectoplasm – wide, clear and translucent, hylaline
– 1/3 of the entire animal
endoplasm – granular
– RBC, leukocytes
 Entamoeba histolytica
E. Morphology
1. Trophozoite
e. nucleus
shape: spherical
size: varies from 4-6μm
location: eccentric position
appears as a finely granular ring
 Entamoeba histolytica
E. Morphology
1. Trophozoite
e. nucleus – structures
i) karyosome – (endosome)
- small, dot-like, central in position and surrounded
by a clear halo
- consists of several granules
- a lining network of fine fibril radiates towards the
periphery of the nucleus – spoke-like radial arrangement
 Entamoeba histolytica
E. Morphology
1. Trophozoite
e. nucleus – structures
ii) nuclear membrane – delicate, lined with a single layer of
uniformly distributed fine chromatin granules
 Entamoeba histolytica
E. Morphology
2. Pre-cystic stage
- smaller than trophozoite; larger than cyst
- varying size: 10–20μm
- colorless
- round or slightly ovoid
- pseudopodial action is sluggish – no progressive movement
- lacks cyst wall
- nuclear morphology is confusing
Entamoeba hitolytica
 Entamoeba histolytica
E. Morphology
B. Cyst
a. size: varies from 10-20μm in diameter
b. shape: round or slightly asymmetrical hyaline bodies;
surrounded by highly refractile cyst wall about 0.5μm thick
 Entamoeba histolytica
E. Morphology
B. Cyst
c. Cytoplasm
i) Chromatoidal bodies
- refractile, sausage-shaped bars with rounded ends
- composed of crystalline ribonucleic acid
- tend to disappear as the cyst matures
 Entamoeba histolytica
E. Morphology
B. Cyst
c. Cytoplasm
ii) glycogen mass
- vacuole
- stains brown with iodine
- believed to represent stored food
- as the cyst matures the glycogen mass gradually disappears
 Entamoeba histolytica
F. Methods of Reproduction
1. Excystation
- process of transformation of cysts to trophozoite
- occurs only when the cysts enter into the alimentary canal of
man
- a quadrinucleate cyst gives rise to 8 amoebulae – each one
capable of developing into a trophozoite
 Entamoeba histolytica
F. Methods of Reproduction
2. Encystation
- not a reproductive process but a means of protection of species
from extinction
- transformation of trophozoites to cysts
- occurs inside the lumen of the intestines of an affected
individual
- lifespan inside lumen – 2 days
- the mature cyst – a quadrinucleate body
– during encystment the nucleus has undergone multiplication
and gives rise to 4 daughter nuclei
 Entamoeba histolytica
F. Methods of Reproduction
2. Encystation
note: excystation and encystation may occur in one and the
same host
- after formation of cysts a transference to another new
host is required
 Entamoeba histolytica
G. Life Cycle
- E. histolytica passess its life cycle only in one host – man
- 2 main phases of development
trophozoite
cyst
pre-cystic form – transitory stage
- mature quadrinucleate cysts – infective stage
 Entamoeba histolytica
F. Methods of Reproduction
3. Multiplication
- occurs only in the trophozoite phase
- trophic forms are exclusively parasitic
- trophozoites grow at the expense of living tissues and multiply
in large numbers
- occurs by binary fission – first the nucleus which divides by a
modified types of mitosis then the cytoplasmic body
 Entamoeba histolytica
G. Life Cycle
- excystation in the cecum or lower ileum (neutral or slighlty
alkaline medium)
- cytoplasmic body retracts and loosens itself from the cyst wall
- vigorous ameboid movement cause a rent to appear on the cyst
wall through which a small mass of cytoplasm comes out,
followed then ultimately by the whole body
 Entamoeba histolytica
G. Life Cycle
Cysts are swallowed

Enter into the alimentary tract

Pass unaltered through the stomach

Cyst wall is digested by action of trypsin in the intestine

 Entamoeba histolytica
G. Life cycle
Excystation in the cecum, liberating a single amoeba with 4 nuclei

A tetranucleate forms 8 amoebulae (metacystic trophozoite)

Young amoebulae being actively motile, invade the tissues and lodge in
submucous tissue of large intestines
 Entamoeba histolytica
G. Life cycle
- Ameba establishes site of infection in the cecal region or may be
swept to the rectosigmoid
- few organisms – reduced chance of establishing foothold in the
intestinal epithelium
- massive and frequent doses acquired in endemic areas are of
epidemiologic importance
 Entamoeba histolytica
H. Pathogenesis
- lesions primarily – intestinal
secondarily – extraintestinal
- intestinal lesions confined to large intestines
- most frequent sites – cecum and rectosigmoid
- less frequent site – ascending colon, rectum, sigmoid or appendix
 Entamoeba histolytica
H. Pathogenesis
- the attachment is mediated by an amebal galactose –
inhibitable adherence lectin
- trophozoites have been shown to activate complement via the
classical and alternative pathways
- the pathogenic activities of E. histolytica depend upon:
1. the resistance of the host
2. the virulence and invasiveness of the amebic strain
3. the conditions in the intestinal tract
 Entamoeba histolytica
H. Pathogenesis
- complications of intestinal amebiasis
1. appendicitis
2. intestinal perforation – commonly in the cecum
3. strictures – confined to the cecum or sigmoid
4. amebic granuloma (ameboma) – may be confused with
neoplastic growth
 Entamoeba histolytica
H. Pathogenesis
1. resistance of the host – resistance depends on innate immunity,
state of nutrition and freedom from infectious and debilitating
disease
2. Virulence – varies with strain
3. Intestinal tract conditions – invasion is facilitated by
carbohydrate diet, physical or chemical injury of the mucosa, stasis
and bacterial flora
 Entamoeba histolytica
H. Pathogenesis
- as the process invades the submucosa
- extends laterally along the axis of the intestines
- tissue destruction is followed by regenerative proliferation of
connective tissue – fibrous thickening
 Entamoeba histolytica
H. Pathogenesis
- during growth trophozoites secrete proteolytic enzyme – tissue
destruction and necrosis
- the early lesion – tiny area of necrosis in the superficial mucosa or
a small modular elevation with minute opening leading to a flask-
shaped cavity
- amoebas are found at the base or periphery of the necrosis
 Entamoeba histolytica
H. Pathogenesis
- the liver abscess is not made of pus
- mixture of sloughed liver tissue and blood
- chocolate-brown and thick consistency
- “anchovy-sauce” like
- can go to spleen, lungs, brain – hematogenous
- subdiaphragmatic abscess, lung abscess
- direct extension
 Entamoeba histolytica
H. Pathogenesis
- Systemic amebiasis – the liver is chiefly invaded – through blood
stream
- dissemination; at times by direct extension
- in the liver, the trophic forms may grow and multiply but
encystation does not occur
- accident on the part of the parasite
- dead end
 Entamoeba histolytica
I. Laboratory Diagnosis
- aim: to demonstrate the presence of E. histolytica in the
material obtained from any particular lesion
 Entamoeba histolytica
I. Laboratory Diagnosis
1. Intestinal amebiasis
a. Asymptomatic patients
1.) Stool examination
b) Microscopic examination
i) natural stool (formed) – cysts
ii) concentration method – cysts
iii) purged stool – motile trophozoite and cysts
iv) sigmoidoscopy – trophozoite
 Entamoeba histolytica
I. Laboratory Diagnosis
1. Intestinal amoebiasis – cases of acute amoebic dysentery
a. Symptomatic patients
1) Stool examination
a) Gross description
- offensive smelling
- dark-brown, semi-fluid
- acidic pH
- admixed with blood, mucous and much fecal matter
 Entamoeba histolytica
I. Laboratory Diagnosis
1. Intestinal amebiasis
a. Symptomatic patients
1) Stool examination
b) Microscopic
- scanty cellular material
- few pus cell
- macrophages
- epithelial cells
- red blood cells are clumped
- Charcot-Leyden crystals
- demonstration of E. histolytica trophozoite
 Entamoeba histolytica
J. Prevention
1. For personal prophylaxis
- boil drinking water
- protect food and drink from contamination by flies, cockroaches
and rats
- avoid eating raw vegetables
- use ice cubes made from boiled water
- personal cleanliness
 Entamoeba histolytica
J. Prevention
2. For community prophylaxis
- effective sanitary disposal of feces
- protection of water supply from fecal pollution
- avoidance of the use of human excrement as fertilizer
- detection and isolation of carriers
 Entamoeba hartmanni
A. Definition
- generally accepted as the name for the ameba formerly
designated as “small race” E. histolytica
- morphologically similar to E. histolytica
- difference only in size
 Entamoeba hartmanni
B. Morphology
Trophozoite: 3-12μm in diameter
Cyst: 4-10μm

* nuclear structures shows same variations as seen in E. histolytica

* trophozoites ingest bacteria but not red blood cells
 Entamoeba hartmanni
C. Pathogenicity
- considered nonpathogenic
- do not treat this infection
 Entamoeba dispar
A. Definition
- capable of causing intestinal lesions in experimental animals
- does not seem able to actually invade tissues
- 9x more prevalent than E. histolytica
 Entamoeba dispar
A. Definition
- Emily Brumot 1925 proposed hypothesis
Species complex
- E. histolytica – invasice species
- E. dispar – morphologically identical non invasive species

- Dispar = different
 Entamoeba dispar
B. Morphology
- because morphologically indistinguishable
- cannot rely on microscopy
- E. histolytica trophozoite ingests RBCs
- regardless of symptoms – presence of E. histolytica-like forms along
with a positive serologic response indicates the presence of true E.
histolytica
- a negative serologic test with E. histolytica like ameba in stools
indicates E. dispar
 Entamoeba dispar
C. Diagnosis
- development of new diagnostic test is essential
- detect antigen in serum by monoclonal antibodies
- detect parasite DNA by nucleotide probes or by PCR amplification
 Entamoeba coli
A. Geographical distribution
- world wide

B. Habitat
- lives in a free state in the lumen of the large bowel
- commensal
 Entamoeba coli
B. Morphology
1. Trophozoite
- size:20-40μm
- sluggishly motile
- cytoplasm not clearly defined
- endoplasm – packed with food vacuoles bacteria and other
substances not red blood cells
- nucleus: large eccentric karyosome surrounded by a broader halo
and coarse chromatin granules lining the thick nuclear membrane
 Entamoeba coli
C. Morphology
2. Cyst
- begins with single nucleus – repeated nuclear division –
octonucleate cyst
- size: 15-20μm in diameter
- in the binucleate stage – large glycogen mass
- chroomatoid bodies – slender filaments or pointed threads
- in mature cysts – no glycogen mass nor chromatoid bodies
 Entamoeba gingivalis
A. Definition
- bears a close morphological resemblance to E. histolytica
found in pyorrheal pockets between teeth and gums and in the
tonsillar crypts
- not really implicated in production of periodontal disease
- most conspicuous under disease conditions – more suitable
environment
 Entamoeba gingivalis
B. Morphology
Trophozoite
- cytoplasm may contain bacteria and occasional red blood cells
- most frequently filled with portions of ingested leukocytes
- only E. gingivalis ingests leukocytes

Cysts – no cysts are formed

 Entamoeba polecki
A. Definition
- first reported as an intestinal parasite of pigs and monkeys
- has been occasionally found in humans
- high incidence in some parts of Papua New Guinea
- most likely route of transmission: pigs-to-humans
 Entamoeba polecki
B. Morphology
Cyst
- single nucleus
- very rarely binucleate or quadrinucleate
- chromatoidal material is abundant
- angular ends and somewhat pointed
- glycogen may be present
- some cysts contain an “inclusion mass”
 Iodamoeba bütschlii
A. Definition
- lives as a harmless commensal in the colon of man
B. Morphology
Trophozoite
- size: 4-20μm in diameter (9-14 μm range)
- sluggishly progressive
- bacteria seen scattered in the cytoplasm
RBCs are not ingested
- nucleus usually not visible
- permanent stain:
nucleus – delicate nuclear membrane
karyosome – large, central position, irregularly rounded, surrounded by a layer of
small chromatin granules
 Iodamoeba bütschlii
B. Morphology
Cyst
- the chromatin granular usually form a crescentic aggregate
between the karyosome and the chromatin granules
- nuclei likened to a basket of flowers:
the karyosome forming the basket;
the linin fibrils the stem;
the granules the blossoms
 Naegleria fowleri
Primary Amoebic Meningo-encephalitis
- “primary” to differentiate it from that caused by E. histolytica
- acute, fulminant, rapidly fatal illness usually affecting children and
young adults
- usually in hot springs, lakes or swimming pools
- from the nasal mucosa – penetrate the cribriform plate and
multiply in the gray matter
 Naegleria fowleri
Primary Amoebic Meningo-encephalitis
- cysts are not formed in the host
- has flagellated stage – bears 2 long flagella
- the ameboid stage usually has a single blunt pseudopodium and
moves rapidly
- transformation of the ameboid form to flagellated form is quick
- nucleus is vesicular with large endosome and peripheral granules
- free living stage: food vacuoles contain bacteria
parasitic stage: food vacuoles filled with host cell debris
 Endolimax nana
Morphology
- size range of trophozoite and cyst similar to Entamoeba hartmanni

Trophozoite
- blunt pseudopodia; sluggish and random
- cytoplasm contains food vacuoles with ingested bacteria
- nucleus: large karyosome, central or eccentric chromatin may
be seen massed against the nuclear membrane without
formation of a distinct karyosome
 Acanthamoeba spp.
A. Definition
- facultative parasite of humans
- biology similar to that of Naegleria but no flagella produced
- cannot tolerate water as hot as Naegleria can
 Dientamoeba fragilis
• Morphology
Traditionally considered a member of the ameba family; but moves
by pseudopodia instead of flagella
- only trophic stage exist
- Size: 12 μm in diameter
- ectoplasm is somewhat differentiated from endoplasm
- a single, broad pseudopodium is usually present
- food vacuoles contain, bacteria, yeast, starch granules, and cellular debris
- About 60% contain 2 nuclei -arrested telophase
- The endosome is eccentric, sometime fragmented or peripheral in the
nucleus