AMOEBA a. E.

coli - Wet mounts – NSS (I2 inhibits movement)

• Pseudopodial locomotion: • LC: (5 stages) - Preservative:
- Rootlike, finger-like or tongue like 1. Mature cyst Ingested – infective stage 1) MIF (Merthiolate iodine formaldehyde)
- Locomotor organelle (ameboid) 2. Stomach (gastric juices on cystic wall) 2) PVA (Polyvinyl alcohol)
- Procurement of food (pseudopodial encirclement) → Holozoic (ingest 3. Small intestine (Excystation) 3) Schaudinn’s fixative
organic matter) 4. Enclosed Metacyst (w/o wall, 8-nuclei) escapes Cyst W #2) & 3) – for permanent staining
Subkingdom: Protozoa 5. Metacyst – cytop. division - Permanent mounts – IH or Trichome stain
Phylum: Sarcomastigophora 6. Metacystic trophozoites 2. Solid or formed feces – CYST (carriers/chronic patient)
Subphylum: Sarcodina 7. Large intestine (cecum) – maturation to troph (feeding st) - Should include a portion of any fleck of mucus
Family: Endamoebidae 8. Mature troph multiply by binary fission adherent to feces or blood
• Strictly parasitic in GIT (alimentary canal) 9. Encystation start – unfavorable condition in cecum - Wet mounts – NSS or I2
• Small; binary fission 10. Undigested food extruded - If only few cysts → conc. → by ZnSO4 centrifugal
• Lack contractile vacuoles 11. Precyst – spherical flotation method → cyst on surface → troph killed
• Most undergo encystation 12. Precyst secrete tough wall → Encystation complete b. Saline-purged specimens (for TROPH)
Genus: 13. Uninucleate cyst - Provide mat for (+) diagnosis if routine fecal exam has
Achromatic 14. Nuclear division been rewarding
Karyosome Chromatin thread 15. Binucleate cyst (Young cyst) - Na2SO4 (Glauber salts) or phosphosoda preferred
(connect K-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 &
Entamoeba
Numerous 18. Passed out w/ feces 3rd bowel movement onto slide → coverslip → examine
Small, center of N Present b. E. histolytica c. Sigmoidoscopy material
granules line NM
• LC: (5 stages) - Scrapings from suspected sites of amoebic ulceration by
1. Mature cyst ingested gentle pressure from long handled curette or loop
Endolimax
2. Stomach - 1/3 of scrapings are from sigmoidorectal area
Thin layer, periph,
Large, blot-like Present 3. Excystation – duodenum - Look for typical lesions
inconspicuous
4. Enclosed metacyst (4 nuclei) escapes its cystic wall - NSS suspension immediately – for motile TROPH
5. Metacyst – cytoplasmic division → 4 metacystic troph - Punch biopsy – fix, section & stain 1st before examining
Iodamoeba Large (1/2 Nuc. 1 layer of
Radiating (Amoebulae) d. Culture – last resort
Diameter) Periendosomal
Rich in chroma granules, no
achromatic 6. Cecum (colonize & feed) → maturation to troph - Study metabolism, pathogenicity & production of
(endosome) periph. chroma
fibrils 7. Mature troph – binary fission antigens for serodiagnosis
8. Start of encystation – unfavorable envi - Inoculum – troph / cyst from feces or mat from c
• Parasitic amoeba (accdg to pathogenicity & habitat):
9. Undigested food extruded out - Medium: dibasic medium of Boeck & Drbohlav (egg slant
A. Nonpathogenic
10. Troph rounds up → Precyst base w/ isotonic overlay = Locke egg serum)
a. Mouth (gingivalis → easy to transfer)
11. Precyst secretes cystic wall - Diamond’s medium TYI-S-33 → reveal E. histo if
b. Intestinal (coli, nana, buetschlii, dispar, hartmanni)
12. Uninucleate cyst (has glycogen mass & chroma. bodies) microscopic exam has failed
B. Pathogenic
13. 2 nuclear divisions → binucleate → quadrinucleate (mature) B. Hepatic Amoebiasis
a. Intestinal (histolytica)
14. Mature cyst a. Presence of Intestinal amoebiasis
• 2 main stages:
- 2 significant sizes for strains: b. Clinical manifestations, inc. WBC, liver function tests
Trophozoite Cyst
a) Large race – ave. diameter: > 10µ, generally virulent (BSP, ALP)
- Chromatoidal bars – crystallized b) Small race - < 10µ (cyst: 5-9µ; troph: 12-15µ) c. Aspiration of abscess – punch/needle biopsy
- Bacteria & food particles,
ribonucleoproteins in cytop. ➢ Commensal, non-pathogenic, “E. hartmanni” - Troph recovered in 1/3 of cases
ingestion (feeding stage)
Only protein source 15. Passed out w/ feces - Content of abscess (choco colored, “anchovy sauce”)
- RBC
- Glycogen vacuole– carb source → mix of sloughed liver tissue & blood or degenerated
- Motility (living state) • Divided into 2:
- Both are lost in mature cyst liver cells, RBC, leukocytes (sometimes)
1. Non-invasive – E. dispar
- Irregular – cytop. extension - Nonpathogenic in man d. X-ray: exhibit damage extent
- Smooth & rounded walls
Most → undergo fixation - In experimental animals: produce intestinal lesions e. Seroimmunologic test:
- Multinuclear (old); young = 1
- Uninuclear - Difficult to distinguish from E. histo (done by culture & 1. Complement fixation
A. Entamoeba biochemical methods) 2. FAT of Goldman
• 3 grps accdg to no. of nuclei in mature cyst: 2. Invasive (?) 3. Indirect hemagglutination (IHA) – more sensitive
1. 1 – E. polecki (pigs, monkey, man) • Strains of E.histo – differing in pathogenicity – distinguished • Contamination thru:
2. 4 – E. histo from nonpathogenic by isozyme analysis 1. Polluted H2O supply – cyst viable in damp soil (8 days), cool
3. 8 – E. coli • Diagnosis: (12 days), H2O (9-30 days), H2O at -4°C (3 months)
• Gingivalis – no encysted form A. Intestinal Amoebiasis 2. Unclean handling of infected indivs (formites, hands, clothes)
• Natural parasites of GIT of vertebrates & invertebrate hosts except a. Stool exam by direct smears & stained mounts 3. Droppings of flies & other insects
E. moshkovskii (sewage H2O & plants) 1. Diarrheic/dysenteric/liquid feces - TROPH - Cysts unchanged in intestine of flies & cockroach
• Inhibit large intestine of vertebrate hosts except gingivalis (man’s - Non-fecal mat: blood & mucus (pick out) - Viable in their feces & vomitus for 48 hrs.
mouth) & bovis (cow mouth) - w/in 30 mins. after voiding (if not, troph disintegrates) - Filth flies (Musca domestica) & cockroaches – mechanical
• Exclusive lumen dwellers exc. E. histo (invade tissue) & E. - do at 3-4 day intervals, not daily vectors of cysts (sticky, bristly appendages carry cyst from
invadens (invade tissue – reptiles) fresh stool; their habit of vomiting & defecating when
 feeding → MoT) FREE-LIVING AMOEBA
4. Human excreta in veggie gardens • Order: Schizopyrenida
5. Carelessness in personal hygiene in children’s asylums, • Fam: Valkamphidae
mental hospitals, prisons & other congested areas • Genus:
• Transmission – sexually transmitted disease (oral – anal route) A. Naegleria
- Human carrier (cyst passers) – sources; show no symptoms a. N. fowleri = N. aerobia
• Pathogenicity - Cause of primary amoebic meningoencephalitis (PAM)
1. Intestinal amoebiasis – localized in colon (colonize & feed) - From lakes, swimming pools (dive - troph → nasal passages
- multiply in crypts → olfactory nerves → cribriform plate → cranium)
- attachment mediated by amoebal galactose or N-acetyl-d- - Uninucleate cyst
galactosamine adherence lectins - 2 forms:
- when ingesting starch granules (rice) 1. Flagellate
- utilize mucous secretions as food ✓ 2 long flagella at one end
- Metabolize anaerobically w/ enteric bacteria ✓ Elongated
- Once they invade tissue – cause lysis ✓ Form pseudopods
- Don’t depend on bacteria – obtain their nourishment thru 2. Ameboid
absorption of dissolved tissue juices ✓ Single blunt lobopodium
- Encystation – not in tissue or outside intestinal lumen → B. Acanthamoeba
specimen taken outside lumen will contain troph only a. Several species (i.e. culbertsoni)
- Affect other organs (liver, brain, lungs, spleen - Can’t tolerate hot H2O as A
- Cause chronic infection of skin or CNS in:
E. coli E. histo 1. Immunocompromised hosts
Nuclear divisions 3 2 2. Agents of keratits (corneal inflammation) w/ contact lenses
Infective stage Mature cyst Young cyst & meningoencephalitis
Metacystic troph 8 4
c. E. 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