Differentiating E. histolytica Trophozoites from the Commensal Amoeba Trophs

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The page below is a sample from the LabCE course Intestinal Protozoan Parasites. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Differentiating E. histolytica Trophozoites from the Commensal Amoeba Trophs

As far as routine microscopic methods, the permanent stained smear is the most important technique for recovery of trophozoites and cysts. The chart below shows some differentiating characteristics between E. histolytica and the non-pathogenic amoebae trophozoites previously mentioned. As you can see, trophozoites are often difficult to distinguish from one another, so it is always best to base diagnoses on the cyst form, if possible.
Nuclear characteristics given are the peripheral chromatin (around the membrane of the nucleus) and the karyosome, which is the central chromatin that appears as a spot or blotch near the center
Images are of trichrome-stained organisms (which is not a differential stain - blue, green, and pink colors vary among structures), and sizes are given, but images are not relative to one another.
Table 4. Differentiating E. histolytica Trophozoites from the Commensal Amoeba Trophs.
OrganismImage NucleusCytoplasm
Entamoeba histolytica
12-60 μm (usual is 15-20 μm)

(7)
Single
Chromatin: finely granular and evenly distributed around periphery of nucleus.
Karyosome: small and central
Finely granular "ground glass" appearance
May contain bacteria or RBC's
E. coli
15-50 μm (usual is 20-25 μm)

(8)
Single
Chromatin: clumped and uneven around periphery
Karyosome: large, may be diffuse, may be eccentric or central
Granular
Usually vacuolated
Can contain bacteria, yeast and other debris
E. hartmanni
5-12 μm (usual 8-10 μm)

(9)
Single
Chromatin: even, may appear as solid ring
Karyosome: small and compact; central or eccentric
Finely granular
Endolimax nana
6-12 μm

(10)
Single
Chromatin: no peripheral chromatin usually seen
Karyosome: large and blot-like, may take up a large portion of the nucleus
Granular, may be vacuolated
Iodamoeba butschlii
8-20 μm (usual 12-15 μm)

(11)
Single
Chromatin: usually no peripheral chromatin
Karyosome: large (occasionally surrounded by refractile granules)
Granular; may be vacuolated
7. Walkowski, Stefan. "Entamoeba histolytica trophozoite." Wikimedia Commons, 4 Oct 2012, https://commons.wikimedia.org/w/index.php?curid=24557823
8. Yasser. "Entamoeba coli trophozoite." Wikimedia Commons, 13 Nov 2008, https://commons.wikimedia.org/wiki/File:Entamoeba_Coli_trophozoite.jpg
9. Osman, Iqbal. "Entamoeba hartmani - Amoebic infections in humans." Wikimedia Commons, 20 Sep 2012, https://commons.wikimedia.org/w/index.php?curid=33628181
10. Centers for Disease Control and Prevention. "Morphological Identification of the Commensal Protozoa of the Human Intestinal Tract Webinar - Endolimax nana." Wikimedia Commons, 30 May 2018, https://commons.wikimedia.org/wiki/File:Morphological_Identification_of_the_Commensal_Protozoa_of_the_Human_Intestinal_Tract.webm
11. Centers for Disease Control and Prevention. "Intestinal (Non-Pathogenic) Amebae - Figure A: Trophozoite of I. buetschlii stained with trichrome." CDC.gov, 29 Oct 2019, https://www.cdc.gov/dpdx/intestinalamebae/index.html