PARASITOLOGY

PRELIMS/LECTURE SECOND SEMESTER

THE AMEBAS

INTESTINAL AMEBAE
● Entamoeba histolytica

COMMENSAL AMEBAE
● Entamoeba hartmanni
● Entamoeba hartmanni
● Entamoeba coli
● Entamoeba polecki
● Entamoeba gingivalis
● Entamoeba nana
● Entamoeba butschlii
CLINICAL MANIFESTATIONS
FREE-LIVING PATHOGENIC AMEBAE
● Naegleria fowleri
● Acanthamoeba spp.

CLINICAL INTESTINAL AMEBAE

Entamoeba histolitica

● Pseudopod-forming non flagellated

○ protozoan parasite
● MOST INVASIVE
● ONLY MEMBER to cause colitis and liver
○ abscess
● Capable of ERYTHROPHAGOCYTOSIS

CYST
● Shape: spherical
● Nucleus: 1-4
○ PC: fine, uniform
○ K: small, central – “bull’s eye”
appearance
● Cytoplasm: with rod-shaped (or cigar shaped)
chromatoidal bars

TROPHOZOITE
● Motility: progressive, unidirectional hyaline
finger-like/blade pseudopod
● Nucleus: single, eccentric
○ Peripheral chromatin (PC): fine uniform
○ Karyosome (K): small, central
● Cytoplasm: fine, granular, ground-glass
appearance, ingested RBCs

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 ● Distinct nuclear membrane line with chromatin
granules
DIAGNOSTIC TEST ● Small karyosome found near the center of the
nucleus

● Microscopic detection (min. of 3 samples

different days)
● Direct Fecal Smear (DFS)
● Formalin Ether Acetate Concentration Test
(FECT)
● Merthiolate Iodine Formalin Concentration Test
(MIFCT)
● Robinson’s and Inoki Stool Culture

TREATMENT & PROGNOSIS

● Metronidazole: for treatment of INVASIVE

AMOEBIASIS
● Diloxanide furoate: for ASYMPTOMATIC CYST
PASSERS
● Percutaneous drainage of liver abscess: patients TROPHOZOITES
who don’t respond to metronidazole ● only one nucleus

Endolimax nana

CYSTS
● Spherical, ovoid, or ellipsoid cysts
● 4 nuclei
● Large, blot-like karyosome (centrally located)
● Absence of PC
● Chromatoid bars are not present

TROPHOZOITES
● Vesicular nucleus
COMMENSAL AMEBAE ● Large, irregularly-shaped karyosome anchored
to the nucleus by achromatic fibrils
● Sluggish, non-progressive motility
Entamoeba hartmanni ● Blot-like in appearance
● Absence of peripheral chromatin is a key feature
CYST in identification of E. nana trophozoites
● Spherical nucleus ● Granular and vacuolated

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 DIAGNOSIS

● mouth scrapings (particularly from the

gingival area)

NOTE:
● lives around the gum line of the teeth in
the tartar and gingival pockets of
unhealthy mouths
Iodamoeba butschlii ● E. gingivalis trophozoites inhabits tonsillar
crypts bronchial mucus
CYSTS ● Nonpathogenic E. gingivalis trophozoites
● Ovoid, ellipsoid, triangular, or other shapes of are frequently recovered in patients
the cysts suffering from pyorrhea alveolaris
● ONLY ONE NUCLEUS ● E. gingivalis was the first ameba
● Large karyosome, eccentric recovered from a human specimen
● Chromatoid bars are absent
● Granulated and vacuolated cytoplasm

TROPHOZOITES
● Sluggish, progressive motility
● Large, chromatin-rich karyosome surrounded by
a layer of achromatic
● globules and anchored to the nuclear membrane
by achromatic fibrils
● peripheral chromatin is absent
● granular and vacuolated cytoplasm
● Infective stage: mature cyst
● Diagnostic stage: cyst and trophozoite in stool

NOTE: Entamoeba coli

● Iodamoeba was coined to describe an
ameba that stains well with iodine. TROPHOZOITE
● Does not undergo typical division ● Motility: nonprogressive,
● Resembles as a basket of flowers in shape sluggish blunt, granular
pseudopod
● Nucleus: 1-8
Entamoeba gingivalis ○ Peripheral
Chromatin
(PC): thicker, irregular
TROPHOZOITES ○ Karyosome (K): large, eccentric
● no cyst stage, and ● Cytoplasm: coarsely granular, vacuolated, dirty
does not inhabit cytoplasm, no RBC
the intestines
● exhibit active
motility
● finely granular cytoplasm
● only ameba that ingests white blood cells

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CYST
● Shape: spherical, oval, triangular
FREE-LIVING PATHOGENIC AMEBAE
● Nucleus: 1-8
○ PC: coarse, granular,
Irregular Naegleria fowleri
○ K: large, eccentric
● Cytoplasm: granular with
pointed ends (splinter-like, ● Causative agent of Primary Amebic
broomstick, jagged ends) Meningoencephalitis (PAM)
Chromatoidal Bodies ● Only ameba with three known morphologic
gorms–ameboid trophozoites, flagellate, and
cysts
DIAGNOSIS
Habitat:
● Thermophilic organisms in hot springs and other
● FECT and Zinc sulfate floatation warm aquatic environments
● Iodine stain
● Swab TROPHOZOITES
● Direct fecal smear ● Vegetative stage
● Granular appearance and a single nucleus
★ NSS (Normal Saline Sol’n) ● Blunt, lobose pseudopodia
- 0.85% ● Directional motility

Entamoeba polecki CLINICAL MANIFESTATION

TROPHOZOITES Primary Amebic Meningoencephalitis (PAM)

● Sluggish, nonprogressive motility
● In diarrheal stools, progressive and TROPHOZOITES
unidirectional ● CSF findings: elevated WBC count with
● Small, central karyosome neutrophilic predominance, high protein, and low
● Fine and evenly distributed PC glucose
● Granular and vacuolated cytoplasm often ● Rare disease that leads to inflammation of the
contains yeast, bacteria and other food particles brain and destruction of the brain tissue
● Sluglike motility of N. fowleri is accomplished by
blunt pseudopodia
● Contains large karyosome, central
● PC is absent
● Granular and often vacuolated cytoplasm

FLAGELLATE FORMS
● Pear-shaped flagellate
● Flagella – assist parasites in locomotion
● Jerky movements or spinning

CYSTS
● Round and have thick cells wall
● Only ONE NUCLEUS
● Large karyosome, central
● Lack peripheral chromatin

CYST DIAGNOSIS
● One nucleus
● Spherical to oval ● Definitive diagnosis of PAM:
● Small central demonstration of characteristic
karyosome trophozoites in the brain and CSF
● Fine and evenly ● Samples of tissue and nasal discharge
distributed PC
● Bacteria-seeded agar culture medium
● Parasites of pigs
exhibit active trophozoites within 24 hours
and monkeys
● PCR and ELISA

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 ★ Kernig’s sign – diagnostic sign for meningitis,
TREATMENT
where the patient is unable to fully straighten his
or her leg when the hip is flexed at 90 degrees
● Amphotericin B in combination with clotrimazole because of hamstring stiffness
● Azithromycin and voriconazole
Acanthamoeba spp.
TROPHOZOITE FORM

It is the etiologic agent of

● Acanthamoeba keratitis (AK)
● Granulomatous Amebic Encephalitis (GAE)

Isolated from: bottled mineral water, soil

swimming pools, deep well water, contact lens
cleaning solution

TROPHOZOITE
FLAGELLATE FORM ● Nucleus: single and large
● Nucleolus: centrally-located, densely staining
● Sluggish and little evidence of progressive
motility
● Cytoplasm: finely granulated w/ large contractile
vacuole
● Acanthapodia: thom-like pseudopodia (for
locomotion): sluggish, polydirectional movement

CYST FORM

CYST
● Round shaped
● Double walled
○ Outer: slightly wrinkled
○ Inner: polyhedral
● Disorganized, granular and sometimes
vacuolated cytoplasm surrounds the Nucleu

NOTE:
● Ameboid trophozoites of N. fowleri are the only
form known to exist in humans
● The cyst form is known to exist only in the
external environment

CLINICAL SYMPTOMS

● PAM occurs when the ameboid trophozoites of

N. fowleri invade the brain, causing rapid tissue
destruction
● Fever, headache, sore throat, nausea, and
vomiting

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 CLINICAL MANIFESTATION

Acanthamoeba Keratitis
● Commonly seen in
○ patients who use
○ CONTACT LENSES

● Cornea shows characteristic annular infiltration

and congested conjunctiva
● Corneal scraping: specimen of choice
● Calcofluor white may be used to stain
Acanthamoeba cysts

Granulomatous Amebic Encephalitis

● Destruction of brain tissue and meningeal
irritation
● Route of invasion: hematogenous
● Incubation period: 10 days
● SPECIMEN OF CHOICE: CSF

DIAGNOSIS

● Acanthamoeba keratitis: epithelial biopsy or

corneal scrapings
● GAE: post mortem
● Trophozoites or cysts in tissues (rare: CSF)
● Non-nutrient agar with Page’s saline overlaid
with Escherichia coli

TREATMENT

Acanthamoeba Keratitis
● Surgical excision of the infected cornea with
subsequent corneal transplantation
● Early recognition of AK coupled with aggressive
combination anti-amebic agents
● Deep Lamellar Keratectomy – Procedure of
choice

Acanthamoeba Mode of transmission

● Aspiration or nasal inhalation of the organisms
● Lower respiratory tract or through ulcers in the
mucosa or skin
● Migrate via hematogenous spread – transported
through bloodstream and invade the central
nervous system (CNS)
● Another route of infection is – eye
○ Contact lens wearers who use
homemade, non sterile saline solutions
○ Experienced trauma to the cornea
● Acanthamoeba castellanii – responsible for most
CNS and eye infections in humans

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