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Amoeba PDF
Amoeba PDF
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
Entamoeba Entamoeba
coli dispar Entamoeba Entamoeba Acanthamoeba Naegleria
gingivalis spp. spp.
histolytica
Entamoeba Entamoeba
hartmanni polecki
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
Intestinal Amoebiasis:
Entamoeba histolytica
Pathogenesis and Clinical Manifestations :
Intestinal Amoebiasis:
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
Entamoeba histolytica
Pathogenesis and Clinical Manifestations :
Entamoeba histolytica
Pathogenesis and Clinical Manifestations :
Extra-Intestinal Amoebiasis:
• Amoebic liver abscess (ALA) or hepatic amoebiasis is the
most common form
The liver is tender and hepatomegaly is present (50%)
Extra-Intestinal Amoebiasis:
• 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
Entamoeba histolytica
Pathogenesis and Clinical Manifestations :
Extra-Intestinal Amoebiasis:
• 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
Entamoeba histolytica
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
Entamoeba histolytica
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
Entamoeba histolytica
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
Entamoeba histolytica
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
Entamoeba histolytica
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
Entamoeba histolytica
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
Entamoeba histolytica
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
Subkingdom: Protozoa
Phylum: Sarcomastigophora
Subphylum: Sarcodina
Superclass: Rhizopoda
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)
Mode of Transmission:
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
Subkingdom: Protozoa
Phylum: Sarcomastigophora
Subphylum: Sarcodina
Superclass: Rhizopoda
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)
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
Geographic Distribution:
• Ordinarily less common than E. coli or E. nana
Geographic Distribution:
• Ordinarily less common than E. coli or E. nana