Parasitology

PARASITIC AMOEBAS (Laboratory)

Parasitology Department’s PPT | September 16, 2022

PARASITIC AMOEBAS 6. Stool Microscopic Examination - Wet Mount with or without contrasting
● Entamoeba Histolytica - Pathogenic/Invasive dye (lodine)
● Entamoeba Coli - Non-Pathogenic 7. Concentration Techniques
● Iodamoeba Buetschlii - Non-Pathogenic o Cyst only identified and Trophozoite are destroyed
● Endolimax Nana - Non-Pathogenic o Removes also other inclusions or background materials
a. Sedimentation
Pathogenic Non-Pathogenic Free-Living
b. Flotation - Cyst rises on the surface (lightweight)

Entamoeba histolytica Entamoeba coli Naegleria

Endolimax Nana Acanthamoeba
Iodamoeba buetschlii Balamuthia
Entamoeba fragilis

*Disease causing Protozoans to Human host

PATHOGENIC AMOEBA
ENTAMOEBA HISTOLYTICA
GENERALITIES
1. Invasive Parasite
2. Located in Fresh Water contaminated by human feces - CDC
3. Causes Amoebic Dysentery
4. Intestinal Disease: Non-Invasive (Asymptomatic - 90% of the cases) or
Invasive
5. Extraintestinal Disease: Liver Abscess (ALA), Lung, and Brain
6.Primary Cutaneous Amoebiasis (Amoebiasis Cutis) - Arises from STOOL EXAMINATION
contaminated wounds. For Entamoeba Histolytica & Other Amoebas
LIFE CYCLE • Trophozoite and Cyst passed in the feces.
• Trophozoite - Diarrheal Stool (Fresh Stool within 30 minutes upon
Collection)
• Cyst - Formed & Soft Stool (Can be examined 1 hour after
collection)
• Both Cyst and Trophozoite - Semi Formed Stool

Diagnostic for Stool Examination

3 stool specimen in different other days for O&P examinations
1. Saline Preparation - Wet Mount (Unstained)/DFS = Visualize Motility (15
sec)
2. lodine Preparation - Best to visualize the Nuclei and Glycogen =
Contrasting
DyeZ
3. Trichome Staining or Iron Hematoxylin Permanent Staining = Morphology

COMPARATIVE SIMILARITIES

METHODS OF DIAGNOSIS
*That is why PCR/Serology is needed
1. Molecular Based Diagnosis
o Preferred Test, High Level of Sensitivity & Specificity
MORPHOLOGY
o Targets DNA for Entamoeba histolvtica in the Stool
1. Identical to E. histolytica (Pathogenic)
2. PCR/DNA
a. E. dispar (Non-Pathogenic)
o Impractical in developing countries
b. E. moshkovskii (Non-Pathogenic)
o Used because of similarity by the morphology of amoebas (pathogenic
2. Other organisms under Entamoeba
vs. non-pathogenic) in the microscope.
3. Culture - Less frequently done but used for research and determination
of pathogenicity.
4. Stool Antigen Detection Tests - Can be used for Initial testing or if
traditional test is negative.

5. Serologic Test (Serum) - IHA (Indirect Hemagglutinin Assay), ElA, IFA, ID =

Useful on EXTRAINTESTINAL/AMOEBIC LIVER ABSCESS = 95% Cases

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 Parasitology
PARASITIC AMOEBAS (Laboratory)
Parasitology Department’s PPT | September 16, 2022

Nuclear Size1/6 Cyst size

*The E. Coli has larger nucleus ¼ cyst size
ENTAMOEBA HISTOLYTICA (TROPHOZOITE)
• Invasive type or Non-Invasive type
INVASIVE FORM: TROPHOZOITE
1. 10-60 um
2. There is presence of NUCLEUS and centrally located KARYOSOME = Evenly
ENTAMOEBA HISTOLYTICA (CYST)
placed Peripheral Chromatin
INFECTIVE FORM: CYST
3. Presence of Ingested RBC - DIAGNOSTIC/PATHOGNOMONIC
1. 10-20 um size
seen for Invasive Trophozoites (Which is NOT SEEN IN OTHER NON-PATHOGENIC
2. Pre-cyst Form - Do not have refractile cyst wall.
AMOEBA)
3. Spherical to slightly oval in shaped
4. Pseudopods = with sharp demarcation with endoplasm and ectoplasm for
4. Cell wall is thin and refractile and the cytoplasm is finely granular
stained specimens
• Other stages (Uni/Bi/Quad)
5. Nuclei - 1 to 4 (Depending on the stage) (Size 1/6 diameter of the cyst)
6. Cytoplasm = With a mass of Glycogen Vacuoles and with Cigar
Shaped Chromatoidal Bodies (Blunt Rounded Ends) = Critical for
differentiation of other cyst

Uninucleate =1 Binucleate = 2 Quadrinucleate = 4

MOTILITY

Progressive, Undirectional, with Finger-like hyaline pseudopods

NON-PATHOGENIC AMOEBA
• Entamoeba Coli
• Iodamoeba Buetschlii
• Endolimax Nana
GENERALITIES
Intestinal Amoeba (Non-Pathogenic) - 5 on this category that can colonize the
GIT (Non-Invasive) but do not consider as pathogenic:
a. Entamoeba coli
b. Entamoeba polecki
c. Entamoeba hartmanni

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 Parasitology
PARASITIC AMOEBAS (Laboratory)
Parasitology Department’s PPT | September 16, 2022

d. Endolimax nana ENTAMOEBA COLI (TROPHOZOITE)

e. Jodamoeba buetschlii MOTILITY
LIFE CYCLE

Sluggish and non-progressive with blunt pseudopods

ENDOLIMAX NANA
• Smallest Amoeba to infect humans
MORPHOLOGY
ENDOLIMAX NANA (CYST)
1. 5-14 um in diameter
2. Spherical to Oval in shape
3. Nuclei and Chromatoidal Bodies - are not discernable in unstained
specimen
4. But for stained - 1-4 small refractile granules representing nuclei
5. Cytoplasm = Clean looking with Ground Glass appearing

ENTAMOEBA COLI
MORPHOLOGY
ENTAMOEBA COLI (CYST)
1. 10-35 um
2. Spherical and oval shape
3. NUCLEI - 1 to 8 sometimes 16 (Size ¼ diameter of the cyst)
4. Presence of Karyosome within the nuclei
5. Mass of Glycogen - On the cytoplasm
6. Chromatoidal Bodies - On the cytoplasm "WHISKBROOM" or Jagged or
Splintered/Pointed ends

ENDOLIMAX NANA (TROPHOZOITE)

1. 6-12um
2. Has atypical nuclei that contain a triangular chromatin mass or band of
chromatin within the nucleus
3. May have single nucleus - Blot like Karyosome
4. Cytoplasm - Granular and highly vacuolated

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 Parasitology
PARASITIC AMOEBAS (Laboratory)
Parasitology Department’s PPT | September 16, 2022

IODAMOEBA BUETSCHLII Glycogen is REDDISH BROWN in Iodine stain

MORPHOLOGY IODAMOEBA BUETSCHLII (TROPHOZOITE)
IODAMOEBA BUETSCHLII (CYST) 1. 8-20 um
1. 6-15 um 2. Motility: Sluggish and Non-Progressive
2. Spherical to Oval in shape 3. Single nucleus with no peripheral chromatin
3. Nucleus – Single, Karyosome is eccentric 4. Large Karyosomal Chromatin
4. Cytoplasm - Has mass of GLYCOGEN occupies 2/3 of the organism =
Diagnostic
5. Very difficult to distinguish with E. Nana

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Parasitology
PARASITIC AMOEBAS (Laboratory)
Parasitology Department’s PPT | September 16, 2022

Reference:
Parasitology Department’s PPT
• CDC
• Medical Parasitology in the Philippines
• Parasitology Laboratory Exercises
• Henry’s Clinical Diagnosis and Management

USE AT YOUR OWN RISK!

ctto

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