Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
Dr. Llanera
January 20, 2014
AMOEBAE
OUTLINE
I. Subkingdom Protozoa
a. Class Lobosea
b. Structure
c. Life Cycle
d. Outbreaks
II. Entamoeba histolytica
a. Amoebiasis
b. Amoebic Colitis
III. Entamoeba hartmanii
IV. Entamoeba coli
V. Entamoeba polecki
VI. Endolimax nana
VII. Iodomoeba Butschii
VIII. Dientamoeba fragilis
IX. Entamoeba gingivalis
X. Naegleria fowleri
XI. Acanthamoeba sp.
SUBKINGDOM PROTOZOA
Phylum Sarcomastigophora
Subphylum Sarcodina
Class Lobosea
Subphylum Mastigophora
Class Zoomastigophora
Phylum Ciliophora
Class Kinetofragminophorea
Phylum Apicomplexa
Class Sporozoa
CLASS LOBOSEA
Intestinal Species
o Entamoeba histolytica
o Entamoeba hartmanni
o Entamoeba coli
o Entamoeba polecki
o Endolimax nana
o Iodamoeba butschlii
o Dientamoeba fragilis (now under Flagellate family)
o Entamoeba dispar
Other Species
o Entamoeba gingivalis
o Acanthamoeba sp.
o Naegleria fowleri
STRUCTURE
Pseudopodia - with pseudopods or finger like structures which extends
for movement
o Lobose
o Crawling motion (not swimming motion)
o E. histolytica active progressive fast movement
o Entamoeba coli sluggish non progressive movement
Nucleus is compact or vesicular with a dark field structure called a
karyosome (endosome or nucleolus)
o In histolytica, the karyosome is smaller and centrally located with
an even peripheral chromatin
o In coli, the karyosome is larger and peripherally located with an
irregular peripheral chromatin
o Only genus entamoeba has peripheral chromatin
o Granules inside the periphery of karyosome is fine in E. histolytica
and coarse in Entamoeba coli
Nucleoplasm
Nuclear membrane
Endoplasm
o With mitochondria
o Food synthesis
o Food vacuoles stored in chromatoidal bodies;
Entamoeba Sp
Entamoeba
histolytica
Entamoeba coli
Karyosome Size
Smaller
Larger
Karyosome Locations
Centrally located
Peripherally
located
Peripheral Chromatin
Even
Uneven
Granules
Fine
Coarse
Chromatoidal Bodies
Movement
Fast, active
Slow, sluggish
Trophozoite Content
RBC
Bacteria, debris
LIFE CYCLE
Person to person transfer (no cystic stage)
Encystation formation of cyst
o Protective - ciliates
o Reproductive flagellates & amoebae
Excystation when cyst becomes trophozites
No intermediate hosts, direct life cycles alternating trophozoites and
cyst
Mature cyst is the infective stage, for E. histolytica mature cyst will
have 4 nuclei and Entamoeba coli would have 8 nuclei
Page 1 of 8
Parasitology 2.3
Figure 1. Life Cycle Mature cyst are ingested. Trophozoites then emerge
from the cyst and invade the colonic wall and reproduce. Mature cyst are
then released in feces
Table 2. Reasons for Amoebic Encystation and Excystation
Encystation
Excystation
pH change (marked)
Dessication of medium
action of the host tissues
Overpopulation
OUTBREAKS
Single or multiple strain
Common in Mental institutions
Polymerase Chain Reaction (PCR) E. histolytica vs. E. dispar
Laredo strain (Laredo, Texas F. H. Connell in 1956)
ENTAMOEBA HISTOLYTICA
AMOEBIASIS: PATHOGENESIS & PATHOLOGY
Sites of colonization
o Intestinal lesion Colon (Most specific is in the cecum followed by
the rectosigmoid)
High requirement for iron
Primary sites of invasion in colon:
o Early flask shaped ulcer
o Late neutrophilic infiltrates
Secondary lesions other levels of the intestine / extraintestinal
VIRULENCE FACTORS
Susceptibility to agglutination by the lectin concanavalin A
Presence of an adhesion lectin that is inhibited by N-acetyl-Dgalactosamine
Ability to adhere to epithelial cells in vitro and to initiate cell-contactdependent cytolysis
Ability to phagocytize cells (E. histolytica demonstrate this thru
ingestion of RBCs erythrophagocytosis).
MOBILITY PATTERN BY STARCH GEL ELECTROPHORESIS
Performed on recent parasite isolates grown in the presence of
bacteria
Virulent strains of E. histolytica grown in axenic culture (w/o bacteria)
retain their zymodeme pattern (isoenzymes). Zymodeme is important
as one of the differentiating factor between E. histolytica and E. dispar.
E. dispar was recognized by Brumpt in 1925 as genetically distinct but
morphologically identical to E. histolytica. With regards to E.
hartmanni, it possesses all the features of E. histolytica but it is smaller
in size.
1913 WALKER AND SELLARDS
Human volunteers ingested cysts.
All became infected but only some developed acute dysentery.
As few as 10 cysts have been shown to produce infection.
Page 2 of 8
Parasitology 2.3
Lungs
SECONDARY LESIONS
Lower colonic segments (rectosigmoid) by regurgitation
Skin
Amebiasis Cutis
o Perianal extension of acute amoebic colitis
o Abdominal wall through rupture or open drainage of a colonic,
appendiceal, or hepatic lesion
o Penis
o Vulvar amoebiasis is less common.
Brain Hematogenous / Arises from or concomitant with liver or lungs
Spleen
Adrenals
Renal System Kidneys, Ureters, Urinary bladders, Urethras
Clitoris
Nasal polyp
Eye
Page 3 of 8
Parasitology 2.3
DETECTION
Specimen: feces, saline-purged or enema specimen, aspirate, surgical
biopsy, necropsy material
Artifacts and confusers WBCs or other parasites
Seroimmunologic diagnosis
o Craig 1928-1931 Complement fixing abilities
o Goldman 1962 1964 Fluorescent antibody
o Indirect hemagglutination Hepatic abscess
o Latex agglutination
o ELISA
o Agar or cellulose acetate diffusion
o Moan hemagglutination test
Trophozoites seen in unstained (but black and white); more seen with
Buffered Methylene blue
Iodine
Buffered
methylene
blue
(if
trophozoites
are seen)
Cysts
Cysts
(occasionally
trophozoites)
Trophozoites
Trophozoites
Page 4 of 8
Parasitology 2.3
ANTIAMOEBICS
Metronidazole kills trophozoites; for E. histolytica, Giardia,
Trichomonas; has metallic taste; do not use with alcohol (disulfiram-like
reactions)
Iodoquinol luminal amoebicide
Diloxanide Furoate luminal, if asymptomatic flatulence, nausea,
rash
Paromomycin Sulfate luminal
Emetine & Dehydroemetine toxic
TREATMENT
Metronidazole 750 mg tid x 5-10 days only used for confirmed cases
of E. histolytica and prophylaxis prior to abdominal surgery
Iodoquinol
Emetine HCl (6% soln) SQ/IM 1 mg/kg BW daily x 5 days (max daily
dose of 60 mg) relieves symptoms > eradicate infection
Dehydroemetine & Emetine with toxic effects on myocardium &
peripheral nerves
DO NOT give the following: Loperamide HCl, Diphenoxylate HCl,
Thephenamil HCl (may produce toxic megacolon in acute ulcerative
colitis).
ENTAMOEBA HARTMANII
small race of E. histolytica
TROPHOZOITES STAGE
Size: 5-12
Motility: Usually nonprogressive
Nucleus
o Number: Not visible in unstained preparations
o Peripheral Chromatin: Fine granules; Evenly distributed;
Uniform in size
o Karyosomal Chromatin: Small; Discrete; Eccentrically located
Cytoplasm
o Appearance: Finely granular
o Inclusions: Bacteria
CYSTIC STAGE
Size: 5-10
Shape: Spherical
Nucleus
o Number: 4 in mature cyst; 1-2 in immature cyst
o Peripheral Chromatin: Fine uniform granules; Evenly
distributed
o Karyosomal Chromatin: Small; Discrete; Centrally located
Cytoplasm
o Chromatid Bodies: Elongated bars with bluntly rounded ends
o Glycogen: Diffuse; Stains reddish-brown with iodine
ENTAMOEBA COLI
TROPHOZOITES STAGE
Size: 15-50
Motility: Sluggish; Non-progressive with blunt pseudopods
Nucleus
o Number: Often visible in unstained preparations
o Peripheral Chromatin: Coarse granules; Irregular in size and
distribution
o Karyosomal Chromatin: Large; Discrete; Eccentrically located
Cytoplasm
o Appearance: Coarse often vacuolated
o Inclusions: Bacteria, Yeasts, etc
ENTAMOEBA POLECKI
Usually seen in hogs and monkeys; rarely diagnosed in man
CYSTIC STAGE
Size: 10-35
Shape: Spherical; Occasionally oval, triangular or another shape
Nucleus
o Number: 8 in mature cyst but there are supernucleate cysts
with 16; 2 in immature cysts
o Peripheral Chromatin: Coarse; Irregularly shaped granules;
Irregularly distributed
o Karyosomal Chromatin: Large; Discrete; Usually eccentrically
located; Occasionally centrally located
Cytoplasm
o Chromatid Bodies: Less in number than E. histolytica;
Splinter-like with pointed ends
o Glycogen: Diffuse; May be a well-defined mass in immature
cysts; Stains reddish-brown with iodine
TROPHOZOITES STAGE
Size: 10-25
Motility: Sluggish; May be progressive in diarrheic stool
Nucleus
o Number: Slightly visible in unstained preparations; May be
distorted by pressure from vacuoles in cytoplasm
o Peripheral Chromatin: Fine granules; Evenly distributed;
Occasionally irregularly arranged; in plaques or crescents
o Karyosomal Chromatin: Small; Discrete; Eccentrically located;
Occasionally large, diffuse or irregular
Cytoplasm
Page 5 of 8
Parasitology 2.3
o
o
CYSTIC STAGE
Size: 9-18
Shape: Spherical or Oval
Nucleus
o Number: 1 to 2
o Peripheral Chromatin: Fine granules evenly distributed
o Karyosomal Chromatin: Small; Eccentrically located
Cytoplasm
o Chromatid Bodies: Many small bodies with angular or pointed
ends; May be oval or rodlike
o Glycogen: Small diffuse masses; Stains reddish-brown with
iodine; A dark area called Incusion Mass is often present;
Inclusion Mass doesnt stain with iodine
ENDOLIMAX NANA
Size: 8-20
Motility: Sluggish; Nonprogressive
Nucleus
o Number: Not usually visible in unstained preparations
o Peripheral Chromatin: None
o Karyosomal Chromatin: Large; Centrally located; Surrounded
by refractive achromatic granules
Cytoplasm
o Appearance: Coarse, granular, Vacuolated
o Inclusions: Bacteria; Yeasts; Etc.
CYSTIC STAGE
Size: 5-10
Shape: Spherical, Ovoid or Ellipsoidal
Nucleus
o Number: 4 in mature cysts; Less than 4 in immature cysts
(rarely seen)
o Peripheral Chromatin: None
o Karyosomal Chromatin: Large; Blot-like; Centrally located
Cytoplasm
o Chromatid Bodies: Granules or small oval masses
o Glycogen: Diffuse; Concentrated mass may be seen in young
cysts; Stains reddish-brown with iodine
CYSTIC STAGE
Size: 5-20
Shape: Ovoid, Ellipsoidal, Triangular or of another shape
Nucleus
o Number: 1 in mature cyst
o Peripheral Chromatin: None
o Karyosomal Chromatin: Large; Eccentrically located; Refractile
achromatic granules on one side; Indistinct in iodine
preparations
Cytoplasm
o Chromatid Bodies: Granular
o Glycogen: Compact well-defined mass; Stains dark brown
with iodine
Trophozoite only
Non-invasive
Page 6 of 8
Parasitology 2.3
Size: 6-12
Motility: Single pseudopodia are multiple leaflike hyaline structures;
motion is active and progressive
Nucleus
o Number: Usually 1 but may be 2
o Rosette-shaped nuclei (Belizario)
o Peripheral Chromatin: None
o Karyosomal Chromatin: Fragmented into 4-8 segments
Cytoplasm
o Appearance: Vacuolated
o Inclusions: Bacteria; Yeast; Starch granules
A. culbertsoni
A. polyphaga
A. castellanii
A. stronyxis
ACANTHAMOEBA
Trophozoite only
First amoeba to be described
Present only in the mouth
Size: 10-20
Motility: Pseudopodia are usually blunt; moderately active and
progressive motility
Nucleus
o Number: One spheroid nucleus
o Peripheral Chromatin: Fine; Evenly distributed
o Karyosomal Chromatin: Coarse
Cytoplasm
o Appearance: Vacuolated
o Inclusions: Food, debris, bacteria
ACANTHAMOEBA CULBERTSONI
Amoebic meningoencephalitis, uveitis and ulceration of cornea
Active trophic forms
o No flagellate form
Resistant cysts resistant to chlorine and can withstand drying
Slow movement of acanthopodia
PATHOGENESIS, PATHOLOGY & SYMPTOMATOLOGY
Purulent leptomeningitis, brain edema, foci of necrosis
Olfactory nerves and lobes not affected
Cerebral hemispheres may be edematous and soft with hemorrhages &
abscesses. (Belizario)
Most affected areas of the brain: posterior fossa, diencephalon,
thalamus, brainstem
On the affected areas, the leptomeninges are opaque with purulent
exudates & vascular congestion. (Belizario)
NAEGLERIA FOWLERI
Free-living amoebo-flagellate
Motile trophozoites
o Amoeboid
o Flagellate (w/ 2 flagella) shed flagella then resume amoeboid
motility and reproduction
Non motile resistant cysts
Flagellate stage enters nasal cavity; where it reverts to amoeboid form
before invading olfactory tissues and the brain
Cysts instilled intranasally are not infective in experimental animals
Page 7 of 8
Parasitology 2.3
MORPHOLOGY
Naegleria
Acanthamoeba
Broad pseudopods
Filamentous pseudopods
(acanthopodia)
Active
Sluggish
Trophozoites
Motility
Flagellate stage
Double walled
None
May have
osteioles
No
Cysts
in
pores
or
APPENDIX
DIAGNOSIS
Amoebae in CSF, scrapings from lesions in cases of corneal or
cutaneous infections; cultures of material from those sources; stained
vaginal smears; purulent discharge from infected ear
TREATMENT
Amphotericin B and sulfadiazine
NAEGLERIA VS ACANTHAMOEBA
Naegleria
Acanthamoeba
Pathogenic : 1 species
Olfactory neuroepithelium
Faster course
Pathogenic: 4 species
Broken or ulcerated skin or eye;
lungs or genitourinary tract
Slow tissue invasion
Granuloma formation
Gradual onset; prolonged chronic
course
Chronically
ill
/
immunosuppressed
Throphozoites (top) and Cysts (bottom): From left to right: (A) E. histolytica,
(B) E. hartmanii, (C) E. coli, (D) E. polecki, (E) Endolimax nana, (F) Iodamoeba
butschii, (G) Dientamoeba fragilis
REFERENCES
Dr. Llaneras lecture & ppt
Philippine Textbook of Medical Parasitology (Belizario)
Page 8 of 8