Amoeba

Francis Anthony V. Soriano, RMT

PAR 313 LEC
College of Medical Technology
Mt. Carmel College

1
 Classification of Protozoan Parasites
• Phylum Sarcomastigophora

Subphylum Sarcodina Acathamoeba Entamoeba dispar

Endolimax nana Entamoeba gingivalis
Entamoeba coli Entamoeba histolytica
Iodamoeba butschlii Naegleria fowleri

Subphylum Mastigophora Chilomastix mesnili Dientamoeba fragilis

Giardia lamblia Trichomonas vaginalis
Trichomonas hominis Trichomonas tenax
Trichomonas vaginalis

Leishmania braziliensis Leishmania donovani

Leishmania tropica Trypanosoma cruzi
Trypanosoma brucei complex

• Phylum Ciliophora Balantidium coli

2
 Classification of Protozoan Parasites (cont.)

• Phylum Apicomplexa Babesia spp

Cryptosporidium hominis
Cyclospora cayetanesis
Isospora belli
Plasmodium spp.
Toxoplasma gondii

• Phylum Microspora Enterocytozon bineusi

Encephalitozoon spp.
Vittaforma cornea
Pleistophora spp.
Brachiola vesicularum
Microsporidium spp.

3
Phylum Sarcomastigophora
Subphylum Sarcodina

GENERAL RULES FOR AMOEBA

• All amoeba are commensal except for Entamoeba histolytica

• With pseudopodium : finger-like structures for movement
• Undergoes ENCYSTATION except for E. gingivalis and
Dientamoeba fragilis.
• Inhabits the large intestine except for E. gingivalis (gums)
• Amoebiasis – presence of amoeba in any part of the body
(exclusively applied to E. histolytica)
• Multiplies through Binary fission

4
 Entamoeba histolytica
Morphologic Forms:
1. Trophozoite – divides through “binary fission”
2. Precyst
3. Cyst – w/ hyaline cystic wall
4. Metacyst

Infective stage : cyst (except for E. gingivalis)

MOT: ingestion of contaminated food and/or water with E. histolytica
cyst
TROPHOZOITE CYST
Vegetative & motile stage Non-motile, non-feeding stage
(feeding stage)
Found in watery, soft or Found in soft to formed stool
semiformed stool
Fragile Resistant to acidic pH
5
Life cycle of E. histolytica

6
Morphologic comparison between E. histolytica and E. coli
A. Trophozoite
Point of E. histolytica E. coli
differentiation
Movement Unidirectional, progressive Sluggish, non-progressive
and non-directional
Shape of Fingerlike Blunted
pseudopodia
Manner or release of One at a time/ explosive Several at a time
pseudopodia
Nucleus uninucleated (central uninucleated (eccentric
karyosome) karyosome)
Inclusions RBC Bacteria, yeast, debris

Cytoplasm Clean looking Dirty looking

size bigger smaller

7
Morphologic comparison between E. histolytica and E. coli

B. Cyst

Point of E. histolytica E. coli

differentiation
No. of nuclei Quadrinucleated More than 4

Chromatoidal bar Sausage, rod, cigar-shaped Broomstick, splinter-like

Nuclear membrane thin thick

8
 TROPHOZOITE
Entamoeba histolytica Entamoeba coli

9
 CYST
Entamoeba histolytica Entamoeba coli

www.cdc.gov

www.studyblue.com 10
 Pathogenesis
Symptoms

• Gradual onset of abdominal pain

• Diarrhea (with or without blood and mucus)
• In children, bloody diarrhea, fever and abdominal pain
• Abscess formation → Amoebic liver abscess

Pathology

Amoebic dysentery vs. Bacilliary Dysentery

Presence of Charcot-Leyden crystals found microscopically in the

stool in cases of Amoebic dysentery.

11
Pathology
• Can cause ulceration “flask-shaped ulcer” in the intestines
(cecum, ascending colon and sigmoid)

Pathogenic determinants / Virulence factors:

1. Lectin : cytoadherence
2. Amoebapores : forms pores
3. Cysteine proteinase : cytopathic for host tissues

www.sayhellotoourlittlefriends.wordpress.com
12
Extraintestinal amoebiasis

• Through the portal vein (liver), trophozoite reach other parts of the
body (liver, brain, lungs, kidneys)

Assymptomatic carriers: cysts becomes unnoticed

Mode of transmission
Ingestion of contaminated food and/or water

Diagnostic Stage: identification of the cyst or trophozoite

Sample for ID: stool (examined within 30 minutes from collection)

13
Laboratory Diagnosis:

1. Direct Fecal Smear

- saline solution : trophozoite motility
- saline + methylene blue : Entamoeba spp. stain blue
- saline + iodine : nucleus of E. histolytica can be
observed
2. Concentration Techniques
- Formalin Ether Concentration Technique (FECT)
- Merthiolate Iodine Formalin Concentration (MIFC)

3.Culture : Robinson’s and Inoki medium

(more sensitive than stool microscopy)

4. Serologic Testing: ELISA, IHAT (can detect past infection), CIE, AGD,
IFAT
5. Molecular Testing : PCR
14
Treatment (Drug of Choice)

Metronidazole : drug of choice for invasive amoebiasis

Diloxanide furoate: drug of choice for asymptomatic cyst passers

Prevention and Control

• Proper hygiene
• Provision for sanitary disposal of human feces
• Improve access to clean and safe drinking water
• Good food preparation practices
• Avoid using “night soil”
• Health education and promotion

15
Non-Pathogenic species

1. Entamoeba coli
• Harmless inhabitant of the colon

2. Entamoeba dispar
• Morphologically similar to E. histolytica, but with different DNA and RNA

3. Entamoeba hartmanni
• Similar to E. histolytica except much smaller and no RBC incusions
• “small-race E. histolytica”

4. Entamoeba polecki
• Parasite of the pigs and monkeys (rarely infect humans)

5. Entamoeba gingivalis
• Can be found in the mouth (gum and teeth surfaces)
• Abundant in cases of oral diseases
• No cyst stage
• Transmission through kissing, droplet spray, sharing utensils
16
Non-Pathogenic species

6. Entamoeba moshkovskii
• morphologically indistinguishable from those of the disease-causing species E.
histolytica and the nonpathogenic E. dispar. Although sporadic cases of human
infection with E. moshkovskii have been reported, the organism is considered
primarily a free-living amoeba.

7. Endolimax nana
• “Smallest amoeba”
• “Cross eyed cyst” – 4 eccentric nuclei

8. Iodamoeba butschlii
• “iodine-cyst” because of its affinity to iodine
• Large glycogen vacuole/ body which stains deeply with iodine
• uninucleated

17
 Endolimax nana cyst

Iodamoeba butschlii cyst 18

FREE LIVING PATHOGENIC AMOEBA

Acanthamoeba

Pathogenesis:
• causative agent of Granulomatous Amoebic Encephalitis (GAE)
- destructive encephalopathy and associated meningeal irritation
• Amoebic keratitis (contact lens users)

MOT:
• use of contaminated swimming pools, deep well, contact lens solutions, etc

Laboratory Diagnosis
• Histopathologic techniques (demonstration of cyst and trophozoites in stool)
→ biopsy
• PCR

Specimen: discharges, exudates, tissue secretions

19
FREE LIVING PATHOGENIC AMOEBA
Naegleria

Pathogenesis:
• Fatal Primary amoebic encephalitis(Naegleria fowleri)

MOT:
• Oral and intranasal routes while swimming in contaminated pools, rivers and
lakes

Diagnosis:
• PCR
• ELISA

Specimen: CSF

Prevention:
• Frequent cleaning
• Chlorination
• Salination
20
Phylum Ciliophora
CILIATES

Balantidium coli
- Causative agent of “balantidiasis or balantidial dysentery”, similar to
amoebic dysentery
- Largest protozoan parasite
- Only parasitic ciliate
- Primarily associated with pigs

Morphology:
- Has trophozoite and cyst stage

Parts:
cytostome : entry of food
cytophage : excretes waste www.southampton.ac.uk
2 disimilar nucleus (macro- and micronucleus)
2 contractile vacuoles

* encystation doesn’t result in increase in nuclei

21
MOT: ingestion of food/water contaminated with B. coli cyst
Pathogenic determinant:
- Hyaluronidase: causes the ulceration
- Ulceration is described as flask-shaped ulcer but with wider
neck

Specimen: stool

Diagnostic stage: cyst and trophozoite

Infective stage: cyst

Treatment: Metronidazole

Prevention: same with other amoeba

22
 REFERENCES
Belizario, V. and De Leon, W. (2004). Philippine Textbook
of Medical Parasitology. Second Edition. University of the
Philippines Manila. Ermita, Manila.

Manahan, E. (2007). Lecture Notes for Parasitology .

University of Santo Tomas. Espana, Manila.

http://www.cdc.gov/parasites/amebiasis/

23