Richard Rupert T.

Vicencio, RMT
DMMC Institute of Health Sciences
▪ Protozoa are unicellular organisms and the lowest form of animal life.

▪ Two morphologic form of ameba life cycle:

1. Trophozoites - the form that feeds, multiplies, and possesses
pseudopods
2. Cysts - the nonfeeding stage

Excystation - the morphologic conversion from the CYST form into the
TROPHOZOITES form, occurs in the ileocecal area of the intestine

Encystation- conversion of TROPHOZOITES to CYST.

▪ Saline wet preparations
▪ Iodine wet preparations
▪ Permanent stains
▪ Separated into two categories:
1. Intestinal
2. Extraintestinal
1. Entamoeba histolytica
▪ Was named by Schaudinn in 1903
▪ Common associated diseases or condition names: Intestinal amebiasis,
amebic colitis, amebic dysentery, extraintestinal amebiasis

▪ Vesicular nucleus, a centrally (or near central) located small

karyosome
▪ varying numbers of chromatin granules adhering to the nuclear
membrane
 1. Entamoeba histolytica
Virulence factors :
1. Gal/Gal Nac lectin
2. Amebapores
3. Cysteine proteinases

*The trophozoites finally erode through the lamina propia and extend
laterally producing the characteristic FLASK SHAPED ULCER
*The most common sites of amebic ulcer are the cecum, ascending
colon, and sigmoid
*Anchovy sauce-like aspirate – appearance of the abscess that is filled
necrotic, proteinaceous debris
1. Entamoeba histolytica
▪ Trophozoites
▪ exhibits rapid, unidirectional, progressive movement with the
help of finger-like hyaline pseudopods.
▪ Nucleus contains a small central mass of chromatin known as a
karyosome (also known as karyosomal chromatin)
▪ Variants of karyosome include eccentric or fragmented
karyosomal material
▪ Peripheral chromatin – surround the karyosome
▪ Contains a fine granular cytoplasm, often referred to as ground
glass in appearance
1. Entamoeba histolytica
1. Entamoeba histolytica
1. Entamoeba histolytica
1. Entamoeba histolytica
▪ Red blood cells (RBCs) in the cytoplasm are considered diagnostic
because E. histolytica is the only intestinal ameba to exhibit this
characteristic
▪ Bacteria, yeast, and other debris may also reside in the cytoplasm,
but their presence, however, is not diagnostic.
1. Entamoeba histolytica
▪ CYST
▪ Young cysts characteristically contain unorganized chromatin material
that transforms into squared or round ended structures call
chromatoid bars
▪ Chromatoid bars - structures that contain condensed RNA material.
▪ glycogen mass - a cytoplasmic area without defined borders that is
believed to represent stored food, is also usually visible in young
cysts.
▪ One to FOUR nuclei are usually present
1. Entamoeba histolytica
1. Entamoeba histolytica
1. Entamoeba histolytica
1. Entamoeba histolytica
1. Entamoeba histolytica
1. Entamoeba histolytica
 1. Entamoeba histolytica
▪ INGESTION OF CYST is the MOT
▪ Other mode of transmission include venereal transmission
through fecal-oral contact and direct colonic inoculation
through contaminated enema equipment.
– gradual onset of abdominal pain and diarrhea with or without blood and
mucus in the stools.
- Amebic liver abscess (ALA) is the most common extra-intestinal
form of amebiasis.
1. Asymptomatic Carrier State
- Three factors, acting separately or in combination, are
responsible for the asymptomatic carrier state of a patient infected with
E. histolytica
a. the parasite is a low-virulence strain
b. the inoculation into the host is low
c. the patient’s immune system is intact
2. Symptomatic Intestinal Amebiasis
- AMEBIC COLITIS, defined as an intestinal infection caused by
the presence of amebas exhibiting symptoms
- AMEBIC DYSENTERY, condition characterized by blood and
mucus in the stool
3. Symptomatic Extraintestinal Amebiasis
- E.histolytica trophozoites that migrate into the bloodstream
are removed by and take up residence in the liver
▪ Microscopic Diagnosis
- Microscopic detection of the trophozoites and cysts in stool
specimens
- a minimum of 3 stools specimens collected in different days
should be examined.
- using DFS, with NSS, visualization of trophozoite motility will
be seen.
- E. histolytica trophozoites with ingested RBC is a diagnostic of
amebiasis.
- CHARCOT LEYDEN crystals can also be seen in the stool.
▪ Concentration methods such as Formalin Ether Concentration TEST
(FECT) and the Merthiolate Iodine Formalin Concentration Test
(MIFC) – are more sensitive than the DFS for detection of cyst.

▪ The following morphologic structures are noted:

1. size of the cyst
2. Number of nuclei
3. Location and appearance of the karyosome
4. Characteristic appearance of the chromatoid bodies
5. Presence of cytoplasmic structures such as glycogen vacuole
▪ Culture
▪ Stool culture using Robinson’s and Inoki medium
▪ Locke’s egg serum

▪ Serology
1. Indirect hemagglutination (IHAT)
2. Counter Immunoelectrophoresis (CIE)
3. Agar gel diffusion (AGD)
4. Indirect fluorescent antibody test (IFAT)
5. Enzyme-linked immunosorbent assay (ELISA)
▪ Metronidazole (Flagyl) is the drug of choice for treatment of invasive
amebiasis
▪ Diloxanide furoate – is the drug of choice for asymptomatic cyst
passer.
- Vaccination
- candidate vaccines molecules which have been most intensely
studied are the:
1. Serine rich E. histolytica protein (SREHP)
2. Adherence lectin (Gal/GalNAc lectin)
3. 29kDa cysteine rich amebic antigen
1. Which of the following structures is (are) typical in trophozoites of
E. histolytica?
A. Single nucleus with a small karyosome
B. Unevenly distributed peripheral chromatin
C. Chromatoid bars
D. Glycogen mass
1. Which of the following structures is (are) typical in trophozoites of
E. histolytica?
A. Single nucleus with a small karyosome
B. Unevenly distributed peripheral chromatin
C. Chromatoid bars
D. Glycogen mass
2. E. histolytica infection is traditionally diagnosed by finding which of
the following?
A. Adult and egg forms of the parasite in a suspected stool sample
B. Trophozoites and/or cysts in a suspected stool sample
C. Larvae in a suspected CSF sample
D. Adult form of the parasite in suspected tissue samples
2. E. histolytica infection is traditionally diagnosed by finding which of
the following?
A. Adult and egg forms of the parasite in a suspected stool sample
B. Trophozoites and/or cysts in a suspected stool sample
C. Larvae in a suspected CSF sample
D. Adult form of the parasite in suspected tissue samples
3. The infective stage of E. histolytica is which of the following?
A. Trophozoite
B. Cyst
3. The infective stage of E. histolytica is which of the following?
A. Trophozoite
B. Cyst
4. Which of the following factors is not responsible for the
asymptomatic carrier state of a patient infected with E. histolytica?
A. Low virulence strain
B. Low inoculation into host
C. Intact patient’s immune syste
D. Patient’s blood type
4. Which of the following factors is not responsible for the
asymptomatic carrier state of a patient infected with E. histolytica?
A. Low virulence strain
B. Low inoculation into host
C. Intact patient’s immune syste
D. Patient’s blood type
5. Which of the following prevention measures can control the spread
of E. histolytica?
A. Drinking tap water
B. Using human feces as fertilizer
C. Boiling water or treat with iodine crystals
D. Practicing unsafe sex
5. Which of the following prevention measures can control the spread
of E. histolytica?
A. Drinking tap water
B. Using human feces as fertilizer
C. Boiling water or treat with iodine crystals
D. Practicing unsafe sex
▪ “small race” E. histolytica because of the many similarities between
the two organisms.
▪ Similar to E. histolytica except that it is much smaller, and does not
ingest red blood cells.
▪ It is also more sluggish in movement.
▪ Immature cysts usually have chromatoidal bars, which may be short
with tapered ends, or thin and bar-like
▪ SIZE accounts for the major difference between the trophozoites and
cysts of Entamoeba hartmanni and Entamoeba histolytica
TROPHOZOITES
- Fingershaped pseudopods exhibiting nonprogressive motility are
standard.
- Trophozoites contain one nucleus that is not typically visible in
unstained preparations.
- peripheral chromatin is usually present as evenly distributed granules
and often has a BEADED appearance
- karyosome may be centrally or eccentrically located
- finely granular cytoplasm of E. hartmanni may contain BACTERIA
- cytoplasm of E. hartmanni does not contain ingested RED BLOOD
CELLS.
▪ The TROPHOZOITE can be differentiated from E. histolytica by the
following features:
1. More vacuolated or granular endoplasm with bacteria and debris.
2. Narrower, less differentiated ectoplasm
3. Broader and blunter pseudopodia
4. More sluggish undirected movements
5. Thicker, irregular peripheral chromatin with a large eccentric
karyosome in the nucleus
▪ An E.coli cyst may be differentiated from E. histolytica by
1. Larger size
2. Greater number of nuclei
3. More granular cytoplasm
4. Splinter-like chromatoidal bodies
6. The trophozoites of E. coli:
A. Have eight nuclei
B. Have a typical nucleus consists of a large, often irregularly shaped
karyosome that is eccentrically located
C. Are characterized by a karyosome that is surrounded by evenly
distributed peripheral chromatin
D. Contain red blood cell inclusions
6. The trophozoites of E. coli:
A. Have eight nuclei
B. Have a typical nucleus consists of a large, often irregularly shaped
karyosome that is eccentrically located
C. Are characterized by a karyosome that is surrounded by evenly
distributed peripheral chromatin
D. Contain red blood cell inclusions
7. Which of the following is not true about E. coli?
A. The parasite is found worldwide.
B. It is considered to be a pathogen.
C. The infection is transmitted through the ingestion of the infected
cyst through contaminated food or drink.
D. The infection can be prevented by adequate disposal of human
feces and good personal hygiene practices.
7. Which of the following is not true about E. coli?
A. The parasite is found worldwide.
B. It is considered to be a pathogen.
C. The infection is transmitted through the ingestion of the infected
cyst through contaminated food or drink.
D. The infection can be prevented by adequate disposal of human
feces and good personal hygiene practices.
▪ Is a parasite of pigs and monkeys.
▪ Rarely, it may infect humans.
▪ It can be distinguished from E. histolytica in that its cyst is
consistently uninucleate.
▪ In stained fecal smears, the nuclear membrane and karyosome are
very prominent.
8. Which of the following statements is not true about the cysts of E.
polecki?
A. Cysts vary in shape from spherical to oval.
B. Cysts contain at least four nuclei.
C. The typical cyst nucleus resembles that of E. histolytica.
D. A diffuse glycogen mass may be present in the cytoplasm of cysts.
8. Which of the following statements is not true about the cysts of E.
polecki?
A. Cysts vary in shape from spherical to oval.
B. Cysts contain at least four nuclei.
C. The typical cyst nucleus resembles that of E. histolytica.
D. A diffuse glycogen mass may be present in the cytoplasm of cysts.
9. Infection with E. polecki is mainly transmitted to humans via which
of the following?
A. Ingestion of E. polecki trophozoite
B. Ingestion of E. polecki cysts
C. Humans do not get infected with E. polecki.
D. Touching an injected pig or monkey
9. Infection with E. polecki is mainly transmitted to humans via which
of the following?
A. Ingestion of E. polecki trophozoite
B. Ingestion of E. polecki cysts
C. Humans do not get infected with E. polecki.
D. Touching an injected pig or monkey
▪ Is characterized by its small size of 6 to 15 um, sluggish movement,
and the characteristic Endolimax nucleus, which has a large irregular
karyosome often described as blotlike in appearance.
▪ Cyst are 6 to 12 um in diameter and are quadrinucleate when mature
▪ The absence of PERIPHERAL CHROMATIN is a key feature that aids in
the identification of E. nana trophozoites.
10. The appearance of an E. nana karyosome is usually which of the
following?
A. Granular
B. Large and round
C. Small and round
D. Blotlike
10. The appearance of an E. nana karyosome is usually which of the
following?
A. Granular
B. Large and round
C. Small and round
D. Blotlike
11. Which of the following statements is true regarding E. nana?
A. E. nana is found primarily in cold regions of the world.
B. E. nana is prevalent in areas in which poor hygiene and substandard
sanitary conditions exist.
C. E. nana is considered as a human pathogen, so treatment is
mandatory.
D. Humans can obtain protection from E. nana via vaccination.
11. Which of the following statements is true regarding E. nana?
A. E. nana is found primarily in cold regions of the world.
B. E. nana is prevalent in areas in which poor hygiene and substandard
sanitary conditions exist.
C. E. nana is considered as a human pathogen, so treatment is
mandatory.
D. Humans can obtain protection from E. nana via vaccination.
▪ Discovered in 1849, E. gingivalis was the FIRST AMEBA recovered from
a human specimen.
▪ Inhabitant of the MOUTH.
▪ Moves quickly and has a numerous blunt of pseudopodia.
▪ Food vacuoles hat contain cellular debris and bacteria are numerous
▪ Lives on the gum and teeth surface, in the gum pockets, and
sometimes in the tonsillar crypts.
▪ NO CYST stage
▪ Transmission is most probably direct through kissing, droplet spray, or
by sharing utensils.
▪ It is particularly important to diagnose E. gingivalis and E. histolytica
correctly because both organisms may be found in the sputum and in
pulmonary abscesses.
▪ E. gingivalis may also be found in the mouths of individuals who
practice good oral hygiene.
▪ E. gingivalis trophozoites have also been recovered in vaginal and
cervical specimens from women who are using intrauterine devices
(IUDs).
12. Which of the following is a unique characteristic of E. gingivalis?
A. The trophozoites exhibit active motility via their pseudopods.
B. There is no known cyst form of this parasite.
C. The trophozoite has a single nucleus with characteristics that
resemble those of E. histolytica.
D. E. gingivalis is the only ameba that may ingest white blood cells.
12. Which of the following is a unique characteristic of E. gingivalis?
A. The trophozoites exhibit active motility via their pseudopods.
B. There is no known cyst form of this parasite.
C. The trophozoite has a single nucleus with characteristics that
resemble those of E. histolytica.
D. E. gingivalis is the only ameba that may ingest white blood cells.
13. E. gingivalis:
A. Is an intestinal ameba
B. Is a pathogen and must be treated with metronidazole
C. Can be found in the mouth and in the genital tract
D. Has a typical amebic life cycle (i.e., trophozoites and cysts)
13. E. gingivalis:
A. Is an intestinal ameba
B. Is a pathogen and must be treated with metronidazole
C. Can be found in the mouth and in the genital tract
D. Has a typical amebic life cycle (i.e., trophozoites and cysts)
▪ The trophozoite is usually 9 to 14 um long
▪ It is identified by its characteristic large vesicular nucleus with a large
endosome surrounded by achromatic granules.
▪ There are no peripheral chromatin granules on the nuclear membrane
▪ The cyst is uninucleated and has a large glycogen body which stains
deeply with iodine
▪ Previous known as the Laredo strain.
▪ First detected in sewage.
▪ Non-pathogenic species that is morphologically indistinguishable from
E. histolytica and E. dispar, but differs from them biochemically and
genetically.
▪ OSMOTOLERANT, able to grow at room temperature (25 to 30 degree
C optimum)
▪ Able to survive at temperatures ranging from 0 to 41 degree C.
14. Iodamoeba bütschlii cysts typically:
A. Contain four nuclei
B. Have a small karyosome in a central position
C. Lack chromatoid bars
D. Lack a glycogen mass
14. Iodamoeba bütschlii cysts typically:
A. Contain four nuclei
B. Have a small karyosome in a central position
C. Lack chromatoid bars
D. Lack a glycogen mass
15. Transmission of I. bütschlii occurs when:
A. The infective cysts are ingested in contaminated food or drink.
B. The infective trophozoites are ingested in contaminated food or
drink.
C. Examining stool specimens from infected individuals
D. An infected mosquito vector bites an unsuspecting human.
15. Transmission of I. bütschlii occurs when:
A. The infective cysts are ingested in contaminated food or drink.
B. The infective trophozoites are ingested in contaminated food or
drink.
C. Examining stool specimens from infected individuals
D. An infected mosquito vector bites an unsuspecting human.
8. Naegleria fowleri
- is the only ameba with three known morphologic forms.
1. ameboid trophozoites
2. flagellate
3. cysts
- is a free-living amebo-flagellate, organism can exist as an
(swimming form)
▪ *Naegleri gruberi is the most commonly studied NON-PATHOGENIC
species that has been utilized in morphologic studies on
differentiation and induction of flagellation as well as studies on
cytoplasmic origin of the flagellar apparatus.
▪ N.philippinensis – isolates from a thermally-polluted stream, an
artificially-heated swimming pool, and from a brain aspirate of a
young patient in the Philippines
2. Flagellated form
- pear-shaped, two whiplike structures that assist select
parasites in locomotion known as flagella extend from the broad end of
the organism.
- motility is jerky or spinning.
- The flagellate trophozoites typically have granular cytoplasm
that often contain vacuoles.
3.Cysts
- have thick cell wall, has only one (1) nucleus, consisting of a
large, centrally located karyosome lacking peripheral chromatin.
- cytoplasm is granular and often contain vacuoles.
▪ The AMEBOID TROPHOZOITES of N. fowleri are the only form known
to exist in humans.
▪ The FLAGELLATE TROPHOZOITES do not divide but rather lose their
flagella and convert back into the ameboid form, in which
reproduction resumes. .
▪ The CYST form is known to exist only in the external environment.
MOT – Swimming in the contaminated water.
▪ AMEBOID TROPHOZOITES enter the human body through the nasal
mucosa and often migrate to the brain, causing rapid tissue
destruction
▪ primarily found in warm bodies of water, including lakes, streams,
ponds, and swimming pools.
▪ In addition to water sources, there have been cases of contaminated
dust containing cyst
▪ some evidence to suggest that sniffing contaminated water may
transmit this ameba.
▪ Asymptomatic - Patients who contract N. fowleri resulting in
colonization of the nasal passages are usually asymptomatic.
▪ Primary Amebic Encephalitis (PAM) - occurs when the ameboid
trophozoites of N. fowleri invade the brain, causing rapid tissue
destruction.
▪ Kernig’s sign - a diagnostic sign for meningitis, where the patient is
unable to fully straighten his or her leg when the hip is flexed at 90
degrees because of hamstring stiffness.
▪ Presence of the trophozoites in the BRAIN and CEREBROSPINAL
FLUID.
▪ Aspirates when introduced into bacteria-seeded agar culture medium,
exhibit active trophozoites within 24 hours of culture.
▪ Naegleria trophozoites can be identified by the presence of blunt,
lobose pseudopodia and directional motility.

▪ Polymerase Chain Reaction (PCR) and Enzyme Linked

Immunosorbent Assay (ELISA) are now used as specific N.fowleri
tests.
▪ Culture
- Balamuth medium
- Nelson medium
▪ Polytene antibiotic amphotericin B – drug of choice for treating PAM
16. The known morphologic forms of Naegleria fowleri are:
A. Ameboid trophozoites, flagellate forms, and cysts
B. Ameboid trophozoites, immature cysts, and mature cysts
C. Eggs, larvae, and adults
D. Flagellate forms, cysts, and larvae
16. The known morphologic forms of Naegleria fowleri are:
A. Ameboid trophozoites, flagellate forms, and cysts
B. Ameboid trophozoites, immature cysts, and mature cysts
C. Eggs, larvae, and adults
D. Flagellate forms, cysts, and larvae
17. The specimen of choice for the recovery of N. fowleri is which of
the following?

a. Sputum
b. Stool
c. Cerebrospinal fluid
d. Urine
17. The specimen of choice for the recovery of N. fowleri is which of
the following?

a. Sputum
b. Stool
c. Cerebrospinal fluid
d. Urine
18. Humans most often contract N. fowleri by which of the following?
A. Swimming in contaminated water
B. Kissing an infected person
C. Practicing unprotected sex
D. Ingestion of contaminated food
18. Humans most often contract N. fowleri by which of the following?
A. Swimming in contaminated water
B. Kissing an infected person
C. Practicing unprotected sex
D. Ingestion of contaminated food
19. The ameboid trophozoites of N. fowleri enter the human body
through all of the following routes except:
A. Entry through the nasal mucosa
B. Inhalation of contaminated dust
C. Sniffing contaminated water
D. Ingesting contaminated food and drink
19. The ameboid trophozoites of N. fowleri enter the human body
through all of the following routes except:
A. Entry through the nasal mucosa
B. Inhalation of contaminated dust
C. Sniffing contaminated water
D. Ingesting contaminated food and drink
20. Practical measures for the control and prevention of N. fowleri
include which of the following?
A. Banning swimming at all times during the summer months
B. Avoidance of chlorinating swimming pools and hot tubs
C. Providing education and awareness in the medical community
D. Avoidance of consuming contaminated food or water prior to
swimming
20. Practical measures for the control and prevention of N. fowleri
include which of the following?
A. Banning swimming at all times during the summer months
B. Avoidance of chlorinating swimming pools and hot tubs
C. Providing education and awareness in the medical community
D. Avoidance of consuming contaminated food or water prior to
swimming
▪ Causative agent of GRANULOMATOUS AMEBIC ENCEPHALITIS (GAE)
and Acanthamoeba keratitis
▪ Is a small free-living ameba characterized by an active trophozoite
stage and a DORMANT cyst stage.
▪ The sluggish motile trophozoites feed on gram-negative bacteria,
blue-green algae or yeasts.
▪ Has been isolated from air, aquaria, bottled mineral water, soil,
swimming pools, deep well water, CONTACT LENS cleaning solutions.
▪ The trophozoite exhibits small, spiny filaments for locomotion known
as acanthapodia.
MOT - aspiration or nasal inhalation of the organisms.
▪ Trophozoites and cysts enter via the lower respiratory tract or through
ulcers in the mucosa or skin.
▪ 2nd route of infection consists of direct invasion of the parasite in the
EYE.
▪ Associated with contaminated saline
▪ the specimen of choice for diagnosing Acanthamoeba spp.
trophozoites and cysts is CSF.
▪ Brain tissue may also be examined.\
▪ CORNEAL SCRAPINGS are the specimen of choice for recovery of
Acanthamoeba infections of the eye.
▪ Calcofluor white may be used to stain Acanthamoeba cysts
▪ Indirect immunofluorescent antibody staining is the technique of
choice for speciating Acanthamoeba.
21. The term acanthopodia refers to:
A. Spinelike pseudopods
B. Hairy projections
C. Double-layer cell wall
D. Large karyosome and no obvious peripheral chromatin
21. The term acanthopodia refers to:
A. Spinelike pseudopods
B. Hairy projections
C. Double-layer cell wall
D. Large karyosome and no obvious peripheral chromatin
21. The specimen of choice for diagnosing Acanthamoeba species
trophozoites and cysts is which of the following?
A. Urine
B. Sputum
C. Cerebrospinal fluid
D. Stool
21. The specimen of choice for diagnosing Acanthamoeba species
trophozoites and cysts is which of the following?
A. Urine
B. Sputum
C. Cerebrospinal fluid
D. Stool
22. Humans may acquire Acanthamoeba species by which of the
following?
A. Aspiration or nasal inhalation of the organisms
B. Direct invasion of the parasites in the eye
C. Swimming or bathing in contaminated water
D. A or B
22. Humans may acquire Acanthamoeba species by which of the
following?
A. Aspiration or nasal inhalation of the organisms
B. Direct invasion of the parasites in the eye
C. Swimming or bathing in contaminated water
D. A or B
23. Infections with Acanthamoeba species are encountered in which of
the following anatomical parts?
A. Eye
B. Large intestines
C. Lungs
D. Liver
23. Infections with Acanthamoeba species are encountered in which of
the following anatomical parts?
A. Eye
B. Large intestines
C. Lungs
D. Liver
24. To prevent infection with Acanthamoeba species, contact lens
wearers should avoid which of the following?
A. Strenuous exercise
B. Foods with high carbohydrate content
C. Wearing clothing made of cotton
D. Using homemade nonsterile saline solutions
24. To prevent infection with Acanthamoeba species, contact lens
wearers should avoid which of the following?
A. Strenuous exercise
B. Foods with high carbohydrate content
C. Wearing clothing made of cotton
D. Using homemade nonsterile saline solutions