4/5/2020

PATHOGENIC PATHOGENIC AND FREE-LIVING

- Entamoeba histolytica - Naegleria fowleri
NON-PATHOGENIC - Acanthamoeba spp.
- Entamoeba dispar - Hartmanella spp.
- Entamoeba hartmanii - Balamuthia mandrillaris
- Entamoeba coli CILIATA
- Entamoeba gingivalis - Balantidium coli
- Entamoeba polecki
- Entamoeba moshkovskii OTHER INT. PROTOZOAN
- Endolimax nana - Dientamoeba fragilis
- Iodamoeba butschlii
- Blastocystis hominis

2 stage of development:
Cyst – Non-motile, Non-Feeding stage, Resistant to acid
Trophozoite – Motile, Feeding stage, Fragile
3 stages of reproduction:
Excystation – development of cyst into trophozoite
Encystation – development of trophozoite into cyst
Binary Fission – mode of reproduction of trophozoite

Entamoeba histolytica Entamoeba histolytica

(Trophozoite) (Cyst)
PARAMETERS DESCRIPTION PARAMETERS DESCRIPTION

Size range: 8-65 um Size range: 8-22 um

Shape: Spherical to round

Motility: progressive, one finger-like
pseudopodia No. of nuclei: One to four
No. of nuclei: One
Karyosome: Small and central
Karyosome: Small and central
Peripheral Fine and evenly
Chromatin: distributed
Peripheral Fine and evenly
Chromatin: distributed Cytoplasm: Finely granular
Cytoplasm: Finely granular Cytoplasmic Chromatoid bars, rounded
Inclusions: ends on young cysts
Cytoplasmic Ingested red blood cell Diffuse glycogen mass in
Inclusions: young cysts

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The only known pathogenic intestinal amoeba. The

range of symptoms varies and depends on two major
factors:
• 🙎 Asymptomatic Carrier state
• 🙎 Symptomatic Intestinal Amebiasis
• 🙎 Symptomatic Extraintestinal Amebiasis

1. Intestinal Amebiasis
a. Amebic colitis / Amebic dysentery : abdominal pain, diarrhea (w/ or w/o mucus & blood)
b. Characteristic ulcer: FLASK SHAPE ULCER
c. Ameboma – mass-like lesion with abdominal pain and dysentery (<1%)
2. Extraintestinal Amebiasis
a. Hepatic amebiasis:
Amebic Liver Abscess: ANCHOVY SAUCE-LIKE EFFUSION
b. Pulmonary amebiasis
c. Cerebral amebiasis
d. Amebic pericarditis
e. Cutaneous amebiasis
f. Genital amebiasis

1. Suppressor factor: neutralizes stomach pH 1. 🙎 Direct Fecal Smear

a. Unstained c. Liquid stool: shows trophozoite
2. Gal/Gal NAc lectin: cytoadherence b. Stained d. Formed stool: shows cyst
3. Amebapores: poreformers 2. 🙎 Liver aspiration biopsy
3. 🙎 Culture: Robinson’s and Inoki medium
4. Cysteine proteinase: Tissue invasiveness 4. 🙎 Serologic Test: ELISA, IHAT, CIE, AGD, IFAT
5. 🙎 Concentration test: FECT, MIFC, FECT, ZnSO4
6. 🙎 Radiographic non-invasive techniques: X-ray/MRI/CT scan
7. 🙎 Molecular techniques (PCR)

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• 🙎 A leading cause of parasitic deaths after only malaria.

• 🙎 Infection occurs is as many as 10% of the world’s population
Disease Drug of choice
• 🙎 Locations at:
Night soil Diloxanide furoate
Asymptomatic cyst passer
Metronidazole
 areas of poor sanitation
Tinidazole
 hospitals for mentally ill Amebic colitis
Metronidazole
 prisons Amebic Liver Abscess Tinidazole
 day care center

• Boiling or treating water with iodine crystal

• Water filtration or chemical treatment
• Proper washing of food products
• Avoiding the use of human feces as fertilizer
• Good personal hygiene
• Protection of food from flies and cockroaches
• Avoidance of unprotected sexual practice

Entamoeba coli
(Trophozoite)
PARAMETERS DESCRIPTION

Size range: 12-55 um

Motility: Non-progressive or Sluggish,

many blunt/rounded pseudopodia
No. of nuclei: One

Karyosome: Large, irregular shape

eccentric
Peripheral Unevenly distributed
Chromatin:
Cytoplasm: Coarse and granulated

Cytoplasmic Vacuoles containing

Inclusions: bacteria often visible

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Entamoeba coli
(Cyst)
PARAMETERS DESCRIPTION
Entamoeba coli
Size range: 8-35 um Geographic areas that have poor hygiene and sanitation
Epidemiology practice
Shape: round to spherical

No. of nuclei: One to eight

No specific drug is needed to treat the infection
Karyosome: Large, irregular shape Treatment
eccentric
Peripheral Unevenly distributed Proper disposal of human feces
Chromatin: Proper personal hygiene practices
Cytoplasm: Coarse and granulated Protection of food and drinks from flies and cockroaches
Prevention and control
Cytoplasmic Diffuse glycogen mass
Inclusions: present in young cyst
thin chromatoidal bars
w/ pointed to splintered
ends in young cyst

Entamoeba hartmanii
(Trophozoite)
PARAMETERS DESCRIPTION

Size range: 5-15 um

Motility: Non-progressive,
finger-like pseudopodia
No. of nuclei: One

Karyosome: small and central

Peripheral Fine and evenly

Chromatin: distributed
Cytoplasm: Finely granular

Cytoplasmic Ingested bacteria may

Inclusions: be present

Entamoeba hartmanii

PARAMETERS
(Cyst)
DESCRIPTION
Entamoeba hartmanii
Size range: 5-12 um
Infections with E. hartmanni are typically asymptomatic
Clinical symptoms
Shape: spherical

No. of nuclei: One to four Geographic areas that have poor hygiene and sanitation
Epidemiology practice
Karyosome: small and central

Peripheral Fine and evenly No specific drug is needed to treat the infection
Chromatin: distributed Treatment
Cytoplasm: Finely granular
Proper disposal of human feces
Cytoplasmic Chromatoidal bars, Proper personal hygiene practices
Inclusions: rounded ends in young Prevention and control Protection of food and drinks from flies and cockroaches
cysts
Diffuse glycogen mass
in young cyst

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Endolimax nana
(Trophozoite)
PARAMETERS DESCRIPTION

Size range: 5-12 um

Motility: sluggish, nonprogressive

blunt pseudopods
No. of nuclei: One

Karyosome: large, irregular, blotlike

Peripheral Absent
Chromatin:
Cytoplasm: granular and vacuolated

Cytoplasmic Bacteria
Inclusions:

Endolimax nana
(Cyst)
Endolimax nana
PARAMETERS DESCRIPTION
Infections are typically asymptomatic
Size range: 4-12 um
Clinical symptoms
Shape: spherical, ovoid, ellipsoid
E. nana is found primarily in warm, moist regions of the world.
No. of nuclei: One to four
Epidemiology Areas in which poor hygiene and substandard sanitary
Karyosome: large, blotlike, usually conditions exist.
central
No specific drug is needed to treat the infection
Peripheral Absent Treatment
Chromatin:
Cytoplasm: granular and vacuolated
Proper disposal of human feces
Proper personal hygiene practices
Cytoplasmic Chromatin granules Prevention and control Protection of food and drinks from flies and cockroaches
Inclusions: nondescript small mass
Diffuse glycogen mass
in young cyst

Iodomoeba butschlii
(Trophozoite)
PARAMETERS DESCRIPTION
Size range: 8-22 um
Motility: sluggish, usually
progressive
No. of nuclei: One
Karyosome: large, eccentric
achromatic
granules may or may
not be present
Peripheral Absent
Chromatin:
Cytoplasm: coarsely granular and
vacuolated
Cytoplasmic Bacteria
Inclusions: Yeast cell
Other debris

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Iodamoeba butschlii
(Cyst) Iodameoba butschlii
PARAMETERS DESCRIPTION
Infections are typically asymptomatic
Size range: 5-22 um Clinical symptoms
Shape: ovoid, ellipsoid, triangular
Found worldwide and has a higher prevalence in tropical
No. of nuclei: One Epidemiology regions
Karyosome: large, eccentric
achromatic granules No specific drug is needed to treat the infection
on one side may be present Treatment
Peripheral Absent Upgrading the personal hygiene and sanitation practice
Chromatin:
Cytoplasm: coarsely granular and Prevention and control
vacuolated
Cytoplasmic well-defined glycogen mass
Inclusions: Granules may be present

Entamoeba gingivalis
(Trophozoite)
Entamoeba gingivalis
PARAMETERS DESCRIPTION Mouth scrapings (gingival areas)
Diagnosis Vaginal and cervical materials
Size range: 8-20 um

Motility: active, varying pseudopod Found in all populations that have been studied for its presence
appearance Epidemiology
No. of nuclei: One
Occur in the mouth and in genital tract but asymptomatic
Karyosome: centrally located
Clinical symptoms Trophozoites are frequently recovered from patients suffering
from pyorrhea alveolaris
Peripheral fine and evenly
Chromatin: distributed No specific drug is needed to treat the infection
Treatment
Cytoplasm: Finely granular
Improved oral hygiene
Cytoplasmic Leukocytes Proper care of teeth and gums
Inclusions: Epithelial cells Prevention and control
Bacteria
Prompt removal of IUD’s in infected patients spontaneously
removes E. gingivalis from the genital tract

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Entamoeba polecki
(Trophozoite)
PARAMETERS DESCRIPTION

Size range: 8-25 um

Motility: Normal stool sluggish non-progressive

Diarrheal Stool Progressive unidirectional

No. of nuclei: One

Karyosome: Small and central

Peripheral fine and evenly

Chromatin: distributed

Cytoplasm: Granular and vacuolated

Cytoplasmic Ingested bacteria

other food particles

Entamoeba polecki
(Cyst)
PARAMETERS DESCRIPTION Entamoeba polecki
Size range: 10-20 um This amoeba is found only in select areas of the world , with
Shape: Spherical or oval highest prevalence occurring in Papua new guinea.
Epidemiology
Considered to be a parasite of pigs and monkeys
No. of nuclei: One

Karyosome: small and central No specific drug is needed to treat the infection
Treatment

Peripheral fine and evenly Protection of food and drinks from flies and cockroaches
Chromatin: distributed Good sanitation and personal hygiene
Prevention and control
Cytoplasm: Granular

Cytoplasmic Chromatoidal bars, angular

Inclusions: pointed ends in young cyst
Glycogen mass in young
cysts , inclusion mass

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Naegleria fowleri
Naegleria fowleri
(Cyst)
(Trophozoite)
PARAMETERS DESCRIPTION
PARAMETERS DESCRIPTION
Size range: 5-22 um
Size range: 8-22 um
Shape: ovoid, ellipsoid, triangular
Motility: Sluglike, blunt No. of nuclei: One
pseudopods
Karyosome: large, eccentric
No. of nuclei: One
achromatic granules
on one side may be
Karyosome: Large and usually
present
centrally located
Peripheral Absent
Peripheral Absent Chromatin:
Chromatin: Cytoplasm: coarsely granular and
vacuolated
Cytoplasm: Granular, usually Cytoplasmic well-defined glycogen mass
vacuolated Inclusions: Granules may be present

• 🙎 Found in warm bodies of water, including lakes, streams,

ponds, and swimming pools.
🙎 Can survive in up to 0.5µg/ml of hyperchlorinated
water
• 🙎 Higher in the summer months of the year
Able to survive in elevated temperatures up to 46˚C
• Can be acquired in contaminated dust

• Infects children to young healthy adults • 🙎 Microscopic examination: Trophozoite

• CSF (wet mount)
• Asymptomatic • Brain (biopsy)
• Kernig’s sign • Tissue (biopsy)
• 🙎 Primary amebic meningoencephalitis (PAM) • nasal discharge
• 🙎Saline wet preparation: CSF
• CSF findings: pleocytosis (high percentage of PMN cells)
• Culture: Balamuth medium, Nonnutrient agar overlayed with
• hypoglycorrhacia Escherichia coli
• elevated protein levels • 🙎 Molecular Techniques: ELISA, PCR

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• Amphotericin B - DOC • 🙎 Posting of off-limit sign around known sources of

• Amphotericin B with rifampin /miconazole contamination
• 🙎 Educating the medical community and public
• 🙎 Swimming pools and hot tubs be adequately chlorinated
• 🙎 Cracks found in the walls of the pools, hot tubs, and baths
should be repaired immediately

Acanthamoeba
(Trophozoite)

PARAMETERS DESCRIPTION

Size range: 12-45 um

Motility: Sluggish, spinelike

pseudopods

No. of nuclei: One

Karyosome: Large

Peripheral Absent
Chromatin:
Cytoplasm: Granular and
vacuolated

Acanthamoeba
(Cyst)
PARAMETERS DESCRIPTION

Size range: 8-25 um

Shape: roundish with ragged edges

No. of nuclei: One

Karyosome: Large and central

Peripheral Absent
Chromatin:
Cytoplasm: Disorganized, granular,
sometimes vacuolated
Cytoplasmic Double cell wall,
Inclusions: smooth polygonal inner cell wall
and outer wrinkeld/jagged cell wall

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• 🙎 Microscopic Examination: • Reported from many countries worldwide both CNS and eye
• CSF (wet mount) infection
• Brain tissue (biopsy) • CNS infection appears in patient who are immunocompromised.
• Corneal scrapings (wet mount)
• Isolated from air, aquaria, bottled mineral water, soil swimming
• 🙎 Culture: Non nutrient agar with E. coli, Proteose-peptone, pools, deep well water, contact lens solutions.
yeast extract, glucose and cysteine (PYGC) containing • Unlike Naegleria fowleri, common Acanthamoeba infection is seen
antibiotics in immunocompromised hosts only.
• 🙎 Molecular techniques: PCR

• 🙎 Granulomatous amebic encephalitis (GAE) • Sulfamethazine

• First documented by Stamm • Itraconazole
• Incubation period: 10 days • Ketoconazole
• 🙎 Acanthamoeba keratitis • Miconazole
• Propamidine isethianate (DOC)
• Rifampin

• Infects both immunocompetent and immunocompromised host

• MOT: Inhalation of cyst from soil, or direct inoculation through skin lesion/cuts
• Skin infection presents painless nodule, then parasite spread hematogenously
and reaches BBB where it penetrates through receptor and proteolytic enzymes.
It can now reproduce through binary fission and cause CNS abrasion.

• Cyst: Round, 10-30 um, single nucleus, multiwalled (3 layers)

• Trophozoite: Broad pseudopods, 30-60 um, large single nucleus with multiple
nucleoli

• Diagnosis: CSF, Skin aspirate. Biopsy, Immunologic testing, and PCR

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Balantidium coli
(Trophozoite)

PARAMETERS DESCRIPTION

Size range: 28-152 um in length

22-123 um wide
Motility: Rotary, Boring

No. of nuclei: Two

Kidney shaped (macronucleus)
small spherical (micronucleus)
Other features One or two visible contractile
vacuoles
Cytoplasm may contain food
vacuoles or bacteria
small cytostome present
Layer of cilia around organism

Balantidium coli
(Cyst)

PARAMETERS DESCRIPTION

Size range: 43-66 um

Number and app. Two

Of nucleus Kidney shaped (macronucleus)
small spherical (micronucleus)

Other features One or two visible contractile

vacuoles in young cyst
Double cyst wall
Row of cilia visible in between
cyst wall layers of young cyst

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Balantidium coli
• Disease: Balantidiasis, balantidial dysentery
• Balantidium coli invades the intestinal epithelium through release of
the enzyme hyaluronidase and creates a characteristic ulcer with a
🙎rounded base and wide neck
• 🙎 Acute diarrhea with mucus and blood, cramps
• Complications include intestinal perforation and acute appendicitis
• 🙎 Microscopic identification of trophozoites and cysts in the
feces using direct examination
• Repeated stools for increase sensitivity
• Concentration techniques
• 🙎 Biopsy from sigmoidoscopy: trophozoite
4/5/2020
• Infective stage: cyst, viable for several weeks
• Human infection results from ingestion of food or water
contaminated with pigs’ fecal material containing Balantidium
coli cysts
• Incubation period: 4-5 days
• Ingested cysts excysts in the small intestine
• Trophozoites inhabit the lumen, mucosa and submucosa of the
large intestine, primarily the cecal region
• usually non-pathogenic commensal in the large intestine
• Many infected individuals are asymptomatic
• Majority of the symptomatic patients complain of diarrhea and
dysentery
• Abdominal discomfort or pain associated with nausea and
vomiting
• Treatment: metronidazole, tetracycline or iodoquinol
• Prevention: proper sanitation, safe water supply and protection
of food from contamination
• Cyst is resistant to environmental conditions and may survive for
long period, they are easily inactivated by heat and 1%
sodium hypochlorite
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1. Vacuolated forms 4. Multiple fission form

• Most predominant
• Large central vacuole (reproductive organelle) and very thick capsule • Arise from vacuolated forms
2. Ameba-like forms • Produce many vacuolated forms
• Active extensions and pseudopodia 5. Cystic form
• Nuclear chromatin: peripheral clumping
• Intermediate stage between vacuolar and pre-cystic • Very prominent and thick osmophilic electron dense wall
3. Granular forms • Infective stage
• Observed from old cultures • Thick walled cyst: responsible for external transmission
• Granular contents develop into daughter cells of the ameba-form when it
ruptures • Thin walled cyst: reinfection within a hosts intestinal tract

Blastocystis hominis
(Vacuolated form)

PARAMETERS DESCRIPTION
• 🙎 Stool is the specimen of choice for the recovery of
Size range: 5-32 um blastocystis
Vacuole: Centrally located
• Concentration technique: increase sensitivity
Fluid-filled structure
consumes almost 90% of organism
• 🙎 Hematoxylin or Trichrome: to differentiate various stages
• 🙎 Culture: Boeck and Drbohlav’s media, Nelson and Jones
Cytoplasm: Appears as a ring around
periphery of organism media

Nuclei: Two or four located in cytoplasm

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• B. hominis infections indicated that they occurred as epidemic • mild to moderate diarrhea
in subtropical countries. • vomiting
• nausea
• fever
• abdominal pain and cramping
• bloating
• flatulence

• Iodoquinol • Proper treatment of fecal material

• Metronidazole (DOC) • Proper hand washing
• Subsequent proper handling of food and water
• Trimethroprim-sulfamethoxazole

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