NCAMD;

DTM&H;DMM

PARASITOLOGY-amoeba

Introduction to Protozoology
Protozoans :
> single cell or unicellular organism
> 2 general stages;
a). Trophozoite or vegetative or feeding stage
b). Cystic stage ( non-motile)
A. Morphology:
vary in size ( B. coli ) ( Plasmodium)
Shape
= assymetrical ( C. mesnili)
= bilateral symmetry ( G.lamblia )
= cuboidal ( B. coli )

Trophozoite stage : >1 nucleus

( B. coli; G. lamblia; & D. fragilis)
no mouth ( cytostome) B. coli

>

B. Cytoplasm:
1. Ectoplasm = outer layer, less granular, more homogenous
2. Endoplasm = food vacuoles ( chromatoidal bars/ glycogen)
Function of cytoplasm:
1. Respiration
2. Protection
3. Ingestion
4. Locomotion ( Pseudopodia, flagella, cilia )

Nucleus = composed of chromatin materials ( reproduction)

Type of Nucleus
1. Vesicular type :a single mass or an aggregate of chromatin
Karyosome = middle of the nucleus or edge of the nucleus,
surrounded by chromatin particles ,
arranged
in a network connecting karyosome with the
nuclear membrane. ( Entamoeba sp)
2. Compact type: large amount of chromatin materials, less
nucleoplasm. ( B. coli)

Other structures:
1. Cytostome or cell mouth
2. Cell anus or cytophage
3. Excretory vacuole
4. Contractile vacuole
5. Nervous system (ciliates & flagellates )kinetoplast
6. Kinetoplast

Physiology of Parasite & Metabolism

Free-living species retain aerobic metabolic pathways
Parasitic species are facultative or obligate anaerobes
> intestinal protozoa ( E. histolytica), survives &
propagate in
low O tension.
> blood & tissue flagellates requires hematin or
growth factor
> Malarial parasites obtain metabolites from
hemoglobin from old
RBC of host & amino acids from plasma of host.

Life cycle
Reproduction = simple binary fission . (Asexual reproduction )
Flagellates : longitudinal axis
Ciliates

: transverse lines

Survival Mechanism of Protozoan;

1. Person to person transfer : ( protozoans with trophic stage
only)
2. Encystation or cyst formation
3. Excystation
4. Sexual phase

Laboratory Diagnosis
1. Stool examination:
>Trophozoites : soft, liquid, or loose consistency
Cyst

: formed stools

a). Saline wet preparations

b). Iodine wet preparation
c). Permanent stain ( I & H )
2. Serologicall testing

AMOEBA

Phylum Protozoa
Subphylum Sarcodina
Class Rhizopodia
Family Entamoebidae
*5 Important Genera :1. Endamoeba
2. Entamoeba
3. Endolimax
4. Iodamoeba
5. Dientamoeba
* Differentiated by the nuclear structures

Entamoeba histolytica
Disease :
> Amebiasis
> Amoebic dysentery
> Amoebic colitis
> Amoebic liver abscess
> amoebic brain abscess
> Cutaneous amebiasis
Geographic Distribution: Prevalent in tropics than in cooler
climates
Transmission : contaminated food & drinks with cyst

E. histolytica
Trophozoite :
Size range

Cyst:
8-65m

Motility = progressive, finger-like

pseudopodia
# of nuclei = one
Karyosome = small & central
Peripheral chromatin = fine &
evenly distributed
Cytoplasm = finely granular
Cytoplasmic inclusions = ingested
RBC

Size range
Shape
round

8- 22 m
spherical to

# of nuclei

14

Karyosome

small & central

Peripheral chromatin = fine &

evenly distributed
Cytoplasm

= finely granular

Cytoplasmic inclusions =
chromatoid bars/ rounded ends,
diffuse glycogen mass

Epidemiology ( E. histolytica )
> 10% worlds population.
> leading cause of parasitic deaths
> subtropical & tropical areas
>also exist in colder climates ( Alaska, Russia & Canada )
> areas of poor sanitation, mental health facility, prisons &
day care
centers
> areas where human waste are used as fertilizer
> homosexual communities
> oral-fecal transmission ( unprotected sex)
> improper treatment of water supplies

Symptomatology :
> Protean or varied depending on site & extent of tissue
damage
IP. Biological = 1-5 days
Clinical

= 4 days to a year

> GI symptoms, dysenteric stools with blood & mucus,

abdominal
cramps, nausea, vomiting
> Liver abscess, upper right abdominal pain, weight loss
> amebic brain abscess= usually fatal . Dx only in necropsy

Pathogenesis & Pathology

Colonization in colon depends on;
1. number of active trophozoites
2. hypermotility
3. suitable enteric bacteria
4. pathogenic index
> small race less than 10micra, nonpathogenic
> large race more than 10 micra , pathogenic

A. Intestinal lesions :
a). Primary lesions = cecum, appendix & ascending colon.
mucosa- muscularis submucosa
Bottle-neck lesion caused by lytic necrosis
b). Secondary lesions = other levels of the intestines &
extraintestinal organs & tissues.( amoebic granuloma )
B. Extra-intestinal lesions :
> hematogenous
> direct extension
> lymphatic route

Flask Shaped Ulcer

Diagnosis :
1. History & clinical manifestations
2. X-ray filling defect with barium meal;
3. Liver scan
4. Proctosigmoidoscopic exam
5. Stool exam
Tx:
1. Metronidazole (Flagyl)
2. Tinidazole ( Fasigyn)
3. Antibiotics ( Tetracycline)

Prevention & control

> Inspection of food service ( handlers, water supply,
& toilet
facilities
> Water treatment regimen
> Proper washing of food products
> avoid use of human feces as fertilizer
> good personal hygiene & sanitation practices
> Food protection from flies & cockroaches
> avoidance of unprotected sexual practices

Entamoeba coli
Trophozoite

Cyst

Size range

12 -55 m

8 -35m

Motility

Non progressive/ blunt

Number of nuclei

one

karyosome

Large, irreg. eccentric

Peripheral chromatin

Coarse, uneven distri

Cytoplasm

coarse

Cytoplasmic inclusions

Bacteria , other debris

Shape

1 -8

Thin chromatoid barssplintered ends, diffuse

glycogen mass
Round to spherical

Differentiation between E. histolytica and E. coli:

E. histolytica

E. coli

RBC in cytoplasm

present

absent

Chromatin body

Cigar shape

Splinter like

Type of pseudopod

Clear, finger like

Blunt

Movement

Directional ( active )

Sluggish

Nucleus ( cyst )

Infective stage

Cyst

Cyst

Location of karyosome

central

peripheral

Pathologic

Yes ( flask-shape ulcer)

No

Entamoeba coli

Endolimax nana
Trophozoite

Cyst

Size range
Motility

5-12
Sluggish, non
progressive, blunt

4-12

# of nuclei
Karyosome
Peripheral
Chromatin

one
Large, irreg, blotlike
absent

1-4
---

Cytoplasm
Cytoplasmic
inclusions

Granular and vacuolated -Bacteria

Chromatin
granules,
glycogen mass

Endolimax nana cyst

Smallest protozoan
Trophozoite: clean cytoplasm
Cyst: 4 nuclei
Clinical specimen: stool

Entamoeba polecki
most common intestinal ameba of humans in Papua,
New Guinea
Trophozoite
Central karyosome
No RBCs inside cytoplasm
Cyst
Single nucleus
Inclusion body
Chromatoidal bars (variable)

E. polecki
Trophozoites

Cyst

Size

8 to 25

10 to 20

Motility(normal stool)

Sluggish nonprogressive

(diarrheal stool) Progressive unidirectional

Number of nuclei

karyosome

Small and central

Small and central

Peripheral chromatin

Fine evenly distributed

Fine evenly distributed

cytoplasm

Granular and vaculated

Granular

Cytoplasmic inclusions

Ingested bacteria

Chromatoid
bars,angular or pointed
ends
Glycogen mass or
inclusion mass

Iodamoeba butschlii
Trophozoite

Cyst

Size range

8 22 m

5- 22m

Motility

Sluggish, progressive

----

# of nucleus

Karyosome

Large, usually cental

Large, eccentric
Refractive achromatic granules (+/-) Achromatic
granules maybe
present

Peripheral
chromatin

absent

absent

Cytoplasm

Coarsely granular & vacuolated

Bacteria, yeast cells other debris

well-defined
glycogen mass
,graules maybe
present

Iodamoeba butschlii

Prominent glycogen cyst

Basket nucleus

Trophozoite is very similar to E. nana

Summary

The typical characteristics that differentiates amoebae:

>size
>shape
>nuclear structure
>cytoplasm appearance
>cytoplasmic inclusions