AMOEBA  Inhibit large intestine of vertebrate hosts except gingivalis (man’s 1.

Diarrheic/dysenteric/liquid feces - TROPH

 Pseudopodial locomotion: mouth) & bovis (cow mouth) - Non-fecal mat: blood & mucus (pick out)
- Rootlike, finger-like or tongue like  Exclusive lumen dwellers exc. E. histo (invade tissue) & E. - w/in 30 mins. after voiding (if not, troph disintegrates)
- Locomotor organelle (ameboid) invadens (invade tissue – reptiles) - do at 3-4 day intervals, not daily
- Procurement of food (pseudopodial encirclement)  Holozoic a. E. coli - Wet mounts – NSS (I2 inhibits movement)
(ingest organic matter)  LC: (5 stages) - Preservative:
Subkingdom: Protozoa 1. Mature cyst Ingested – infective stage 1) MIF (Merthiolate iodine formaldehyde)
Phylum: Sarcomastigophora 2. Stomach (gastric juices on cystic wall) 2) PVA (Polyvinyl alcohol)
Subphylum: Sarcodina 3. Small intestine (Excystation) 3) Schaudinn’s fixative
Family: Endamoebidae 4. Enclosed Metacyst (w/o wall, 8-nuclei) escapes Cyst W #2) & 3) – for permanent staining
 Strictly parasitic in GIT (alimentary canal) 5. Metacyst – cytop. division - Permanent mounts – IH or Trichome stain
 Small; binary fission 6. Metacystic trophozoites 2. Solid or formed feces – CYST (carriers/chronic patient)
 Lack contractile vacuoles 7. Large intestine (cecum) – maturation to troph (feeding st) - Should include a portion of any fleck of mucus
 Most undergo encystation 8. Mature troph multiply by binary fission adherent to feces or blood
Genus: 9. Encystation start – unfavorable condition in cecum - Wet mounts – NSS or I2
10. Undigested food extruded - If only few cysts  conc.  by ZnSO4 centrifugal
Achromatic
11. Precyst – spherical flotation method  cyst on surface  troph killed
Karyosome Chromatin thread
12. Precyst secrete tough wall  Encystation complete b. Saline-purged specimens (for TROPH)
(connect K-NM)
13. Uninucleate cyst - Provide mat for (+) diagnosis if routine fecal exam has
Entamoeba
14. Nuclear division been rewarding
Numerous
Small, center of N Present 15. Binucleate cyst (Young cyst) - Na2SO4 (Glauber salts) or phosphosoda preferred
granules line NM
16. Quadrinucleate cyst - After purgation  discard earlier fecal evacuations 
17. Octanucleate cyst (Mature cyst) pipette sedimented el. of mucus & tissue cells from 2 nd &
Endolimax
18. Passed out w/ feces 3rd bowel movement onto slide  coverslip  examine
Thin layer, periph,
Large, blot-like Present b. E. histolytica c. Sigmoidoscopy material
inconspicuous
 LC: (5 stages) - Scrapings from suspected sites of amoebic ulceration by
1. Mature cyst ingested gentle pressure from long handled curette or loop
Iodamoeba Large (1/2 Nuc. 1 layer of
Radiating 2. Stomach - 1/3 of scrapings are from sigmoidorectal area
Diameter) Periendosomal
achromatic 3. Excystation – duodenum - Look for typical lesions
Rich in chroma granules, no
fibrils 4. Enclosed metacyst (4 nuclei) escapes its cystic wall - NSS suspension immediately – for motile TROPH
(endosome) periph. chroma
5. Metacyst – cytoplasmic division  4 metacystic troph - Punch biopsy – fix, section & stain 1st before examining
 Parasitic amoeba (accdg to pathogenicity & habitat): d. Culture – last resort
(Amoebulae)
A. Nonpathogenic - Study metabolism, pathogenicity & production of
6. Cecum (colonize & feed)  maturation to troph
a. Mouth (gingivalis  easy to transfer) antigens for serodiagnosis
7. Mature troph – binary fission
b. Intestinal (coli, nana, buetschlii, dispar, hartmanni) - Inoculum – troph / cyst from feces or mat from c
8. Start of encystation – unfavorable envi
B. Pathogenic - Medium: dibasic medium of Boeck & Drbohlav (egg slant
9. Undigested food extruded out
a. Intestinal (histolytica) base w/ isotonic overlay = Locke egg serum)
10. Troph rounds up  Precyst
 2 main stages: - Diamond’s medium TYI-S-33  reveal E. histo if
11. Precyst secretes cystic wall
Trophozoite Cyst 12. Uninucleate cyst (has glycogen mass & chroma. bodies) microscopic exam has failed
- Chromatoidal bars – 13. 2 nuclear divisions  binucleate  quadrinucleate (mature) B. Hepatic Amoebiasis
- Bacteria & food
crystallized ribonucleoproteins 14. Mature cyst a. Presence of Intestinal amoebiasis
particles, ingestion
in cytop. protein source - 2 significant sizes for strains: b. Clinical manifestations, inc. WBC, liver function tests
(feeding stage)
Only - Glycogen vacuole– a) Large race – ave. diameter: > 10µ, generally virulent (BSP, ALP)
- RBC
carb source b) Small race - < 10µ (cyst: 5-9µ; troph: 12-15µ) c. Aspiration of abscess – punch/needle biopsy
- Motility (living
- Both are lost in  Commensal, non-pathogenic, “E. hartmanni” - Troph recovered in 1/3 of cases
state)
mature cyst 15. Passed out w/ feces - Content of abscess (choco colored, “anchovy sauce”)
- Irregular – - Smooth & rounded  Divided into 2:  mix of sloughed liver tissue & blood or degenerated
cytop. extension  walls 1. Non-invasive – E. dispar liver cells, RBC, leukocytes (sometimes)
Most
undergo fixation - Multinuclear (old); - Nonpathogenic in man d. X-ray: exhibit damage extent
- Uninuclear young = 1 - In experimental animals: produce intestinal lesions e. Seroimmunologic test:
A. Entamoeba - Difficult to distinguish from E. histo (done by culture & 1. Complement fixation
 3 grps accdg to no. of nuclei in mature cyst: biochemical methods) 2. FAT of Goldman
1. 1 – E. polecki (pigs, monkey, man) 2. Invasive (?) 3. Indirect hemagglutination (IHA) – more sensitive
2. 4 – E. histo  Strains of E.histo – differing in pathogenicity – distinguished  Contamination thru:
3. 8 – E. coli from nonpathogenic by isozyme analysis 1. Polluted H2O supply – cyst viable in damp soil (8 days), cool
 Gingivalis – no encysted form  Diagnosis: (12 days), H2O (9-30 days), H2O at -4°C (3 months)
 Natural parasites of GIT of vertebrates & invertebrate hosts except A. Intestinal Amoebiasis 2. Unclean handling of infected indivs (formites, hands, clothes)
E. moshkovskii (sewage H2O & plants) a. Stool exam by direct smears & stained mounts 3. Droppings of flies & other insects
- Cysts unchanged in intestine of flies & cockroach
 - Viable in their feces & vomitus for 48 hrs.
- Filth flies (Musca domestica) & cockroaches – mechanical FREE-LIVING AMOEBA
vectors of cysts (sticky, bristly appendages carry cyst from  Order: Schizopyrenida
fresh stool; their habit of vomiting & defecating when  Fam: Valkamphidae
feeding  MoT)  Genus:
4. Human excreta in veggie gardens A. Naegleria
5. Carelessness in personal hygiene in children’s asylums, a. N. fowleri = N. aerobia
mental hospitals, prisons & other congested areas - Cause of primary amoebic meningoencephalitis (PAM)
 Transmission – sexually transmitted disease (oral – anal route) - From lakes, swimming pools (dive - troph  nasal passages
- Human carrier (cyst passers) – sources; show no symptoms  olfactory nerves  cribriform plate  cranium)
 Pathogenicity - Uninucleate cyst
1. Intestinal amoebiasis – localized in colon (colonize & feed) - 2 forms:
- multiply in crypts 1. Flagellate
- attachment mediated by amoebal galactose or N-acetyl-d-  2 long flagella at one end
galactosamine adherence lectins  Elongated
- when ingesting starch granules (rice)  Form pseudopods
- utilize mucous secretions as food 2. Ameboid
- Metabolize anaerobically w/ enteric bacteria  Single blunt lobopodium
- Once they invade tissue – cause lysis B. Acanthamoeba
- Don’t depend on bacteria – obtain their nourishment thru a. Several species (i.e. culbertsoni)
absorption of dissolved tissue juices - Can’t tolerate hot H2O as A
- Encystation – not in tissue or outside intestinal lumen  - Cause chronic infection of skin or CNS in:
specimen taken outside lumen will contain troph only 1. Immunocompromised hosts
- Affect other organs (liver, brain, lungs, spleen 2. Agents of keratits (corneal inflammation) w/ contact lenses
& meningoencephalitis
E. coli E. histo
c. Nuclear divisions 3 2 E.
Infective stage Mature cyst Young cyst
Metacystic troph 8 4
hartmanni – LC, morpho & appearance identical to E. histo
except size (like E. nana)
Troph: - don’t ingest RBC
- motility less vigorous than histo
- Nuc: like coli in char. of its chromatin & karyosome
Cyst: - glycogen mass
- chromatoidal bodies (short w/ tapered ends; rice-grain
shaped or thin, bar-like)
d. E. dispar
B. Endolimax
a. E. nana - Same stage & LC as E.coli
C. Iodamoeba
a. I. buetschlii
Amoebic Dysentery Bacillary Dysentery
Gross Appearance Gelatinous mixture of Mucopurulent mass
blood, mucus & feces streaked w/ blood
Amt Copious Small
Odor Offensive (fishy) Inoffensive
Color Dark red Bright red
Reaction Acidic Alkaline
Microscopic
Ghost cells
None 95% degenerated
(WBC remnants)
Macrophages Rare Present
Never clumped,
RBC Clumped
discrete
Charcot-Leyden
crystals (in stools Present Absent
w/ parasitic infxn)
Bacteria Numerous Nil to none
Pus cells Scanty numerous