THE AMEBAS

Entamoeba histolytica
PARAMETERS
TROPHOZOITE CYST
Size Range 8 to 65 um 8 to 22 um
Motility/Shape Progressive, finger-like pseudopodia Spherical to round
Number Of Nuclei One One to four
Karyosome Small and central Small and central
Peripheral Chromatin Fine and evenly distributed Fine and evenly distributed
Cytoplasm Finely granular Finely granular
Cytoplasmic Inclusions Chromatoid bars, rounded ends in young cysts
Ingested RBC
Diffuse glycogen mass in young cysts

LABORATORY DIAGNOSIS
- Wet preparation and permanent staining techniques
- Spx collected from a sigmoidoscopy procedure and hepatic abscess material
- Special medium TYI-S-33 supports E. histolytica in culture
- Antigen tests (ELISA, IHA, Gel diffusion precipitin, and Indirect immunofluorescence)
o Only helpful in cases of extraintestinal infections
 Entamoeba hartmanni
PARAMETERS
TROPHOZOITE CYST
Size Range 5 to 15 um 5 to 12 um
Motility/Shape Non-progressive, finger-like pseudopods Spherical
Number Of Nuclei One One to four
Karyosome Small and central Small and central
Peripheral Chromatin Fine and evenly distributed Fine and evenly distributed
Cytoplasm Finely granular Finely granular
Cytoplasmic Inclusions Chromatoid bars, rounded ends in young cysts
Ingested bacteria may be present
Diffuse glycogen mass in young cysts

LAB DIAGNOSIS
- Differentiated to E. histolytica only by measuring its size using an ocular micrometer
o Sizes of both parasites overlap
 Entamoeba coli
PARAMETERS
TROPHOZOITE CYST
Size Range 12 to 55 um 8 to 35 um
Motility/Shape Non-progressive, blunt pseudopods Round to spherical
Number Of Nuclei One One to eight
Karyosome Large, irregular shape, eccentric Large, irregular shape, eccentric
Peripheral Chromatin Unevenly distributed Unevenly distributed
Cytoplasm Coarse and granulated Coarse and granulated
Diffuse glycogen mass present in young cysts
May display nuclei to opposite ends of the cyst
Cytoplasmic Inclusions Vacuoles containing bacteria often visible
Thin chromatoid bars with pointed to splintered ends
in young cysts

LAB DIAGNOSIS
- Stool examination is the method of choice
- Presence of this parasite suggests ingestion of contaminated food or drink
- Parasite is not considered as being pathogenic
 Entamoeba polecki
PARAMETERS
TROPHOZOITE CYST
Size Range 8 to 25 um 10 to 20 um
Motility/Shape Sluggish, non-progressive (normal stools)
Spherical or oval
Progressive, unidirectional (diarrheal stools)
Number Of Nuclei One One
Karyosome Small and central Small and central
Peripheral Chromatin Fine and evenly distributed Fine and evenly distributed
Cytoplasm Granular and vacuolated Granular
Chromatoid bars, angular or pointed ends in young
Ingested bacteria cysts
Cytoplasmic Inclusions
Other food particles Glycogen mass in young cysts
Inclusion mass

LAB DIAGNOSIS
- Trophozoites and cysts may be diagnosed by examining stool samples
 Endolimax nana
PARAMETERS
TROPHOZOITE CYST
Size Range 5 to 12 um 4 to 12 um
Motility/Shape Sluggish, non-progressive, blunt pseudopods Spherical, ovoid, ellipsoid
Number Of Nuclei One One to four (four most common)
Karyosome Large, irregular, blotlike Large, blotlike, usually central
Peripheral Chromatin Absent Absent
Cytoplasm Granular and vacuolated Grnaular and vacuolated
Chromatin granules
Cytoplasmic Inclusions Bacteria Non-descript small mass
Diffuse glycogen mass in young cysts

LAB DIAGNOSIS
- Stool examination
 Iodamoeba butschlii
PARAMETERS
TROPHOZOITE CYST
Size Range 8 to 22 um 5 to 22 um
Motility/Shape Ovoid, ellipsoid, triangular
Sluggish, usually progressive
Other shapes
Number Of Nuclei One One
Large, usually central
Large, eccentric, achromatic granules which may be
Karyosome Refractive achromatic granules may or may not be
present on one side
present
Peripheral Chromatin Absent Absent
Cytoplasm Coarsely granular and vacuolated Coarsely granular and vacuolated
Bacteria
Well-defined glycogen mass
Cytoplasmic Inclusions Yeast cells
Granules may be present
Other debris

LAB DIAGNOSIS
- Stool examination is the method of choice
- Iodine wet preps for the identification of I. butschlii cysts
o Glycogen mass picks up the iodine stain making it recognizable
o Glycogen mass remains unstained after trichrome staining
 Entamoeba gingivalis
PARAMETERS
TROPHOZOITE CYST
Size Range 8 to 20 um
Motility/Shape Active, varying pseudopod appearance
Number Of Nuclei One
Karyosome Centrally located
Peripheral Chromatin Fine and evenly distributed NO KNOWN CYST
Cytoplasm Finely granular
Leukocytes
Cytoplasmic Inclusions Epithelial cells
Bacteria

LAB DIAGNOSIS
- Diagnosis is best by examining mouth scrapings (gingival area)
- Material from tonsillar crypts and pulmonary abscess as well as sputum may also be examined
- Vaginal and cervical material may also be examined in the vagina and in the cervix
 Naegleria fowleri
PARAMETERS
TROPHOZOITE CYST
Size Range 8 to 22 um 9 to 12 um
Motility/Shape Sluglike, blunt pseudopods Round and have thick cell walls
Number Of Nuclei One One
Karyosome Large and usually centrally located Large, centrally located
Peripheral Chromatin Absent Absent
Cytoplasm Granular, usually vacuolated Granular, and often vacuolated

LAB DIAGNOSIS
- Microscopic examination of CSF is the method of choice of N. fowleri trophozoites
- Saline and iodine wet preps of CSF are recommended
- Tissue samples and nasal discharge may also be examined
- Trophozoites show a trailing effect when placed on agar plates that have been previously inoculated with gram-negative bacilli
 Iodamoeba butschlii
PARAMETERS
TROPHOZOITE CYST
Size Range 12 to 45um 8 to 25 um
Motility/Shape Sluggish, spinelike pseudopods Roundish with ragged edges
Number Of Nuclei One One
Karyosome Large Large and central
Peripheral Chromatin Absent Absent
Cytoplasm Granular and vacuolated Disorganized, granular, sometimes vacuolated
Other features - Double cell wall – smooth inner and jagged outer

LAB DIAGNOSIS
- CSF is the specimen of choice
- Brain tissue may also be examined
- Corneal scrapings are the specimen of choice for recovery of Acanthamoeba spp. Infections of the eye
o May be cultured on non-nutrient agar plates seeded with gram-negative bacteria (viable strain of E. coli)
o Bacteria serve as food for the parasites
o Produce set of marks (tracks) on the agar
- Calcofluor white may be used to stain Acanthamoeba cysts in corneal scrapings
- Indirect immunofluorescent antibody staining is the technique of choice for speciating Acanthamoeba
 INTESTINAL SPECIES EXTRAINTESTINAL SPECIES
Entamoeba histolytica
Entamoeba hartmanni
Entamoeba gingivalis
Entamoeba coli
Naegleria fowleri
Entamoeba polecki
Acanthamoeba species
Endolimax nana
Iodamoeba butschlii

Stool CSF Mouth Corneal

Scrapings Scrapings
E. histolytica X
E. coli X
Endolimax nana X
Entamoeba gingivalis X
Acanthamoeba spp. X X
Naegleria fowleri X

Metronidazole Amphotericin
Paromomycin Furamide Iodoquinol Rifampin Miconazole Sulfamethazine
(Flagyl) B
E. histolytica X X X X
E. polecki X X
Acanthamoeba spp. X X X
Naegleria fowleri X X X

PATHOGENIC NON-PATHOGENIC

Entamoeba hartmanni
Entamoeba histolytica
Entamoeba coli
Entamoeba polecki
Endolimax nana
Naegleria fowleri
Iodamoeba butschlii
Acanthamoeba species
Entamoeba gingivalis
THE FLAGELLATES
Giardia intestinalis
PARAMETERS
TROPHOZOITE CYST
Size Range 8 to 20 um long 8 to 17 um long
5 to 16 um wide 6 to 10 um wide
Motility/Shape Pear-shaped, teardrop
Ovoid
Falling leaf
Appearance Bilaterally symmetrical -
Two (immature cyst)
Nuclei Two ovoid-shaped, each with a large karyosome Four (mature cyst)
Central karyosomes
4 pairs:
One pair (anterior)
Flagella -
One pair (posterior)
Two pairs (central, extending laterally)
Cytoplasm - Retracted from cell wall
Median bodies
Two median bodies
Two (immature)
Other structures Two axonemes
Four (fully mature)
Sucking disk
Interior flagellar sturctures
 Chilomastix mesnili
PARAMETERS
TROPHOZOITE CYST
Size Range 5 to 25 um long
5 to 10 um long
5 to 10 um wide
Motility/Shape Pear-shaped
Lemon-shaped, with clear hyaline knob
Stiff, rotary, directional
Nuclei One with small central or eccentric karyosome One with large central karyosome
Four:
Flagella Three extending (anterior end) -
One extending (posterior from cytostome region)
Prominent cytostome (1/3 to ½ body length) Well-defined cytostome located on one side of the
Other structures
Spiral groove nucleus

LAB DIAGNOSIS
- Freshly passed liquid stools can only reveal trophozoites
- Formed stool reveal only cysts
- Semiformed stools contain trophozoites and cysts
- Iodine wet preps often demonstrate organism’s features clearly
 Dientamoeba fragilis
PARAMETERS
TROPHOZOITE CYST
Size Range 5 to 10 um
Motility/Shape Irregularly round
Progressive, broad hyaline pseudopodia
Two NO KNOWN CYST
Nuclei Each consisting mass clumps
4 to 8 chromatin granules
Other structures Bacteria-filled vacuoles common

LAB DIAGNOSIS
- Stool examination for the presence of trophozoites is the method of choice
- Multiple samples may be necessary to rule out its presence
- May be difficult to find and much less identified because of its ability to blend well in the background
- Conventional and RT-PCR have been used to diagnose this organism
o RT-PCR is the most sensitive of all diagnostic methods
 Trichomonas hominis
PARAMETERS
TROPHOZOITE CYST
Size Range 7 to 20 um long
5 to 18 um wide
Motility/Shape Pear-shaped
Nervous, jerky
Nuclei One with small central karyosome
Three to five anterior
NO KNOWN CYST
Flagella One posterior extending from the posterior of
undulating membrane
Axostyle extends beyond the posterior end of the body
Full body length undulating membrane
Other structures
Conical cytostome cleft in anterior region ventrally
located opposite the undulating membrane

LAB DIAGNOSIS
- Stool examination is the method of choice
 Enteromonas hominis
PARAMETERS
TROPHOZOITE CYST
Size Range 3 to 10 um long 3 to 10 um long
3 to 7 um wide 4 to 7 um wide
Motility/Shape Oval; sometimes half-circle
Oval, elongated
Jerky
One to four
Binucleated and quadrinucleated nuclei located at
Nuclei One with central karyosome
opposite ends
Central karyosomes
Four:
Flagella Three directed anteriorly -
One directed posteriorly
Other structures None None

LAB DIAGNOSIS
- Stool examinations is the method of choice
- Difficult to identify because of its small size
 Retortomonas intestinalis
PARAMETERS
TROPHOZOITE CYST
Size Range 3 to 7 um long 3 to 9 um long
5 to 6 um wide Up to 5 um wide
Motility/Shape Ovoid
Lemon-shaped or pear-shaped
Jerky
One with small central karyosome One (anterior-central region) with central karyosome
Nuclei Ring of chromatin granules may be in nuclear May be surrounded by a delicate ring of chomatin
membrane granules
Flagella Two (anterior) -
Cystostome extending halfway down body length Two fused fibrils (bird’s beak) in the anterior nuclear
Other structures Well-defined fibril border opposite the nucleus in the region
anterior end Only visible in stained preparations

LAB DIAGNOSIS
- Stained stool sample is the best sample to examine
- Difficult to identify because of its small size
- Small number of diagnostic features may sometimes not stain well enough to be recognized
 Trichomonas tenax
PARAMETERS
TROPHOZOITE CYST
Size Range 5 to 14 um long
Motility/Shape Oval, pear-shaped
One, ovoid nucleus
Nuclei Consists of vesicular region filled with chromatin
granules
Five (all originating anteriorly)
Flagella 4 extending anteriorly NO KNOWN CYST
1 extending posteriorly
Undulating membrane extending 2/3 of body length
Thick axostyle curves around nucleus extending beyond
Other structures body length
Small anterior cytostome opposite the undulating
membrane

LAB DIAGNOSIS
- Mouth scrapings (specimen of choice)
- Microscopic examination of tonsillar crypts and pyorrheal pockets often yields typical trophozoites
- Tartar between teeth and gingival margin of the gums are the primary areas of the mouth may potentially harbor this organism
 Trichomonas vaginalis
PARAMETERS
TROPHOZOITE CYST
Size Range Up to 30 um long
Motility/Shape Ovoid, round or pear-shaped
Rapid, jerky
Nuclei One, ovoid, nondescript
All originating anteriorly:
NO KNOWN CYST
Flagella 3 to 5 extending anteriorly
1 extending posteriorly
Undulating membrane extending half of body length
Other structures Prominent axostyle that often curves around nucleus
Granules may be seen along axostyle

LAB DIAGNOSIS
- May be recovered in spun urine, vaginal discharges, urethral discharges, and prostatic secretions
- Saline wet preps are preferred in many cases
- Phase contrast microscopy, Pap smears, fluorescent stains, monoclonal antibody assays, enzyme immunoassays and cultures
- Affirm VPIII – a DNA-based assay
- InPouch TV culture system – can be used with vaginal swabs (women), urethral swabs (men), urine and semen sediments
o Requires incubation time and takes up to 3 days before a result is determined
 Stool Mouth Vaginal/Urethral
Scrapings Swabs
Giardia intestinalis X
Chilomastix mesnili X
Dientamoeba fragilis X
Enteromonas hominis X
Retortamonas intestinalis X
Trichomonas hominis
Trichomonas tenax X
Trichomonas vaginalis X

PATHOGENIC NON-PATHOGENIC
Chilomastix mesnili
Giardia intestinalis Trichomonas hominis
Dientamoeba fragilis Enteromonas hominis
Trichomonas vaginalis Retortamonas intestinalis
Trichomonas tenax

INTESTINAL SPECIES EXTRAINTESTINAL SPECIES

Giardia intestinalis
Dientamoeba fragilis
Trichomonas vaginalis
Chilomastix mesnili
Trichomonas tenax
Trichomonas hominis
Enteromonas hominis
Retortamonas intestinalis

Flagyl Tinidazole Nitazoxanide Iodoquinol Tetracycline

Giardia intestinalis X X X
Dientamoeba fragilis X X
Trichomonas vaginalis X
RELATIVE SIZES OF HELMINTHS