Free Living Amoeba

Maricelle D. Manlutac, RMT, MPH, MLS (ASCPi)

Faculty
Free Living Amoeba

 True pathogen and opportunistic

 Acquired while diving and swimming in warm water or improper

chlorinated pool

 Inhabits CNS, cornea, and skin

 Naegleria fowleri; Acanthamoeba castellani; Balamuthia mandrillaris

Acanthamoeba spp.
Naegleria fowleri
 3 Stages

a) an amebic/amoeboid trophozoite (the only stage

that exist in human)

b) a biflagellate/ ameboflagellate form

c) cyst
Amoeboid Trophozoite
 Rounded / elongated

 The infective stage

 Single nucleus

 Divide by binary fission.

 Can be transformed into flagellate

and cyst form

 Found in CSF and tissue

Flagellate form
 Elongated / pear shaped

 Anterior nucleus

 Two free flagella

 Found in water at 27-37 ̊C

 Non feeding & non dividing

 Never present in tissue

 Cyst form

 Rounded with single nucleus

 Thick double cyst wall

 Found in soil

 Never in tissue
Naegleria fowleri
 True pathogen

 Known as “Brain-eating amoeba”

Pathology: Primary Amoebic Meningoencephalitis

Dx based on trophozoites in the brain and CSF

**Naegleria philippinensis
Acanthamoeba spp.

 Associated with contact lens cleaning solutions

 Trophozoite with “Thorn-like” appendages

Pathology:

a. Keratitis – Dx by epithelial biopsy or corneal scrapings

 A. castellani, A. culbertsoni, A. hutchetti, A. polyphaga, A.
rhysoides

b. Granulomatous encephalitis (GAE)

Naegleria fowleri
Acanthamoeba spp.
Prevention
 Public education

 Proper chlorination of pools

Babesia spp.

 Hemosporidian parasites

 Definitive host is animal (Deer)

 Causative agent of Babesiosis / Piroplasmosis

 Man infected by:

1. Bite of IH
a) Hard Tick- Ixodes scapularis
b) Soft Tick – Ornithodoros erraticus

2. Blood Transfusion

 More pathogenic - B. bovis; B. equi; Less pathogenic- B. bigemia; B. caballi

 Babesia bigemia- Red water fever in cattle

Lab Diagnosis

 Giemsa Stained smear

 Presence of tetrad formation or “Maltese cross” form

 Serologic test
 IFA

 Intraperitoneal inoculation of hamster

 PCR
 Gold standard
Babesia VS Plasmodia
1. Absence of pigments in the schizogony phase
 [+] Plasmodium
 [-] Babesia

2. Lack of growing trophozoite and gametocyte

 [+] Plasmodium
 [-] Babesia

3. Ability to grow in hamster blood

 [-] Plasmodium
 [+] Babesia

4. Number of invading trophozoite w/in a single RBC

 [ ≤ 3 ] Plasmodium
 [ ≤ 12] Babesia

5. Number of chromatin dots

 [ ≤ 2] Plasmodium
 [ ≤ 3] Babesia
Phylum: Apicomplexa
Class : Conoidasida
Subcl. : Coccidia
Order : Eucoccidiorida
Suborder: Eimeriorina (Tissue & Intestinal Coccidia)
Family : Eimeriidae Cryptosporiidae Sarcocystidae

 Genus : Isospora Cyclospora Cryptosporidium Sarcocystis Toxoplasma

 Species: belli cayetanensis parvum hominis suihominis gondii

hominis
Coccidians

 Obligate intracellular protozoan

 Secretory organelles ( rhoptries, micronemes, and polar rings)

Coccidian life cycle:

1. Sexual cycle (SPOROGONY) Oocyst

2. Asexual cycle (SCHIZOGONY) Merozoites

3. Gametogony Gametocytes
Cryptosporidium spp.

 Infect human and most mammals (AIDS/

immunocompromised)

 Associated with watery Frothy diarrhea with oocyst

 IS and DS is oocyst containing sporozoites measuring 4-6µ in

diameter

Types of oocyst:

a. Thin walled- affect other RBC resulting in autoinfection and

chronicity

b. Thick walled- fecal oral route contamination

** Infections by C. felis, C. meleagridis, C. canis, and C. muris

have also been reported
Lab Dx
 DFS – examination of
oocyst
 Sheather’s sugar
flotation
 FECT
 Modified acid fast
 Oocyst – red-pink
doughnut shaped in blue
background

 Sputum sample – Figure 1. Modified Acid-fast Staining showing

pulmonary involvement Cryptosporidium Oocysts
Cyclospora cayetanensis

 Food- and waterborne coccidia

 Previously classified as cyanobacterium-like body (CLB)

Lab Dx:

a. DFS

b. Modified Acid fast staining

c. Fluorescent microscopy – blue-green circles

d. PCR
Isospora belli

Pathology:

 Villous atrophy commonly associated with malabsorption Syndrome

 Associated with Cystoisosporiasis

Lab Dx

a. DFS

b. Modified Acid fast stain

c. String capsule
Toxoplasma gondii

 Intracellualr parasite

 Definitive host is the house cat and certain other Felidae

 IS includes:

a) Tachyzoites- in initial and acute stages; mother-fetus transfusion

b) Bradyzoites – form cyst; meat infected animals

**both can be transferred by organ transplant

a) Oocyst
Toxoplasma gondii

MOT:

a. Fecal oral ( Cat feces)

b. Eating infected meat (Bradyzoites)

c. Organ transplant (Tachyzoites and Bradyzoites)

d. Transplacental during acute infection (Tachyzoites)

Diagnosis of Toxoplasma
infection:

 Isolation of parasites from blood or other body fluids (GIEMSA)

 Sabin-Feldman methylene blue dye test (Live organisms)

 Serologic testing is the routine method of diagnosis

 Detection of parasite genetic material by PCR, especially in detecting

congenital infections in utero.
Hydrocephaly and Microcephaly
Sarcocystis spp.
MOT: consumption of raw meat with sarcocyst

Pathology: Sarcocystosis

 Invasive form- vasculitis and myositis

 Intestinal form

Lab Dx:

a. DFS- sporocyst

b. Muscle biopsy- definitive dx

c. PAS

d. PCR