Protozoa.

Sarcomastigophora.
Sarcodina. Amoebida.
 Learning Objectives
AMOEBA
PATHOGENIC AMOEBAE
Intestinal species
Extra intestinal
Life cycle
Infective stage
Lab diagnosis (protozoan diagnostic stage)
NON-PATHOGENIC AMOEBAE
 Class – RHIZOPODA (Amoebae)
AMOEBA- Protozoa: Single-celled eukaryotic microorganisms belonging to
kingdom protista are classifed as Protozoa (Greek Protos: first; zoon: animal).
• The single protozoal cell performs all functions (reproduction,digestion,
respiration,excretion
• Exhibit movements by cilia, flagella, pseudopodia.
Classification: Genus – Entamoeba . Species – histolytica
• Intestinal species: 1.Entamoeba histolytica 2 .E.hartmanni 3. E.coli
4.E.polecki. 5.Endolimax nana. 6.Iodamoeba butschlii.
• Extra intestinal -1.Entamoeba gingivalis.
• 2. Acanthamoeba sp.
• 3.Naegleria fowleri.
 General Features
Phylum – protozoa
• Most of the protozoa are completely nonpathogenic but few may cause major
diseases such as malaria, leishmaniasis, and sleeping sickness.
• Protozoa like Cryptosporidium parvum and Toxoplasma gondii are being
recognized as opportunistic pathogens in patients afected with human
immunodefciency virus (HIV) and in those undergoing immunosuppressive
therapy.
• Protozoa exhibit wide range of size (1–150 µm), shape, and structure; yet all
possess essential common features.
• Basic structure- protoplasm enclosed in cell membrane. Protoplasm
differentiated into cytoplasm& Nucleus.
AMOEBA
E
Amoebae of the alimentary canal:
The most important of these is E.
histolytica, which causes intestinal
and extraintestinal amoebiasis.
Amoebae are also present in the
mouth.

Potentially pathogenic free-living amoebae: Several species of saprophytic

amoebae are found in soil and water.
Two of these, Naegleria and Acanthamoeba are of clinical interest because
they can cause eye infections and fatal meningoencephalitis.
Structure
• The typical protozoan cell is bounded by a trilaminar unit membrane, supported
by a sheet of contractile fibrils enabling the cell to move and change in shape.
Cytoplasm
It has 2 portions:
Ectoplasm: Outer homogeneous part that serves as the
organ for locomotion and for engulfment of food by
producing pseudopodia .
It also helps in respiration, discharging waste material, and
in providing a protective covering of cell.
Endoplasm: the inner granular portion of cytoplasm that
contains nucleus . The endoplasm shows number of structures
—the golgi bodies, endoplasmic reticulum, food vacuoles,
and contractile vacuoles.

Contractile vacuoles serve to regulate the osmotic pressure

E. histolytica causes intestinal and extra intestinal amoebiasis
Intestinal amoebiasis -PATHOGENESIS
Lumen dwelling amoeba do not cause any illness .They causes disease only
when they invade the intestinal tissues .
10 % -symptomatic
90% -asymptomatic
Entamoeba histolytica
• Entos = inside + amoibes = changing; histos = tissue + luein = to dissolve
• Was first described by Losch in 1875 after being isolated in Russia from a
patient with dysenteric tool.
• Only amoeba to have definite pathogenic potential is Entamoeba
histolytica, a major cause of bloody diarrhea (dysentery) and extra-
intestinal invasive disease in humans.
• 2nd most common parasitic cause of mortality (after malaria), accounting
for an estimated 55,000 deaths each year.
• Risk factors
– Young age, pregnancy, corticosterioid use, alcoholism, malignancy,
malnutrition etc.
 Entamoeba histolytica
Type of protozoan: Amoeba.
• Disease caused by protozoan: Amoebic dysentery. (Amoebiases)
It occurs in three stages:

Trophozoite, precyst cyst

 › E. histolytica
Diagnostic stage: Cyst or trophozoite in stool.
• E. histolytica associated with intestinal & extra
intestinal infection.
• E. histolytica inhabits large intestine.
• The other Species are important because the
may be confused with E. histolytica
• They are transmitted by Feco-oral route.
 Entamoeba histolytica
Trophozoite (Trophe = Food + Zoon = animal)
• Trophozoite is the vegetative or growing stage of the parasite . It is the
only form present in tissues.
• It is amoeboid in shape .
• 20-30µm in size (though it could range from 10- 60µm)
• Actively motile in fresh stool with short hyaline pesudopodia
 Trophozoite

• The trophozoites from acute dysenteric

stools often contain phagocytosed erythrocytes.
This feature is diagnostic as phagocytosed red
cells are not found in any other commensal
intestinal amoebae.
• The trophozoites divide by binary fission in every 8 hours.
• Trophozoites survive upto 5 hours at 37°C and are killed by drying, heat, and
chemical sterilization. Therefore, the infection is not transmitted by
trophozoites. Even if live trophozoites from freshy-passed stools are ingested,
they are rapidly destroyed in stomach and cannot initiate infection present in
tissues.
Precystic Stage
•Trophozoites undergo encystment in the intestinal lumen.
•Pre- cysts → mature cysts (10x20µm)
•Encystment does not occur in the tissues nor in feces outside the body.
•It then secretes a highly retractile cyst wall ( which makes it highly resistant to gastric
juice and unfavorable environmental conditions) around it and becomes cyst

Cystic Stage

› An early cyst has one nucleus → divides into

two or four nuclei (metacyst) on maturation.

Infective stage: Cyst.

Development in man (small
intestine)
Excystation: In small intestine, the
cyst wall gets lysed by trypsin and a
single tetranucleated trophozoite
(metacyst) is liberated which
eventually undergoes a divisions to
produce eight small metacystic
trophozoites
• Metacystic trophozoites are
carried by the peristalsis to
ileocecal region of large intestine
and multiply by binary fission, and
then colonize on the mucosal
surfaces and crypts of the large
intestine.
Development in man (large intestine)
• Encystation: After some days, when the
intestinal lesion starts healing and patient
improves, the trophozoites transform into
precysts then into quadrinucleated cysts which
are liberated in feces. Encystation occurs only in
the large gut. Cysts are never formed once the
trophozoites are excreted in stool Factors that
induce cyst formation include food deprivation,
overcrowding, desiccation, accumulation of
waste products, and cold temperatures
Mature quadrinucleated cysts released in feces
can survive in the environment and become the
infective form.
Immature cysts and trophozoites are some times
excreted, but get disintegrated in the
environment.
Life cycle of E. histolytica / E. dispar .

Mature cyst stained with 4%

Lugol solution (100×
magnification).
b) Mature cyst without
staining (100×).
c) Trophozoite observed with
differential interference
contrast (DIC) (100×).
d) Trophozoites of E.
histolytica species with
phagocyted erythrocytes
(DIC 40×).
• The metacystic trophozoites penetrate the columnar
epithelial cells of crypts of Liberkuhn
• Penetration is facilitated
• by motility of the trophozoites
• tissue lytic enzyme –histolysin
• amoebic lectin -mediate adherence
• Sometimes invasion remains superficial and heal
spontaneously.
• More often, the amoeba penetrates to submucosal
layer and multiplies rapidly causing lytic necrosis
and thus forming an abscess →ulcer (do not extend
deeper than submucosal layer)
Virulence factors
Trophozoites of E.histolytica interact with host through a series of sof
steps
• 1 Adhesion of target cell, phagocytosis and cytopathic effect
• 2 E.histolytica induces both Humoral and cell mediated immune
responses.
• 3 In many circumstances lumen dwelling Amoeba may be
asymptomatic
• 4 Causes disease only when invade the Intestine
 Pathogenesis & pathology

• E . Histolytica can effect almost all tissues of human

body .
• Hydrolytic enzymes : proteolytic destruction of
tissues
• Amoebic lectin : mediates in the adherence of
amoebae to intestinal mucosa
• Cysteine proteniase which assists the organism in
digesting the extracellular matrix and invading
tissues. Amoebic cystine proteinase, inactivates
complement factor C3 is an important virulence
factor.
• Ionophore like protein : causes leakage of ions i.e..,
Na , K, Ca from target cell .
Intestinal Amoebiasis
› Causes disruption of the epithelium and the depletion of the mucus → followed by
formation of ulcers.
› Lesions commonly develop
– Caecum
– Rectum
– Ascending colon
– Sigmoid colon
Rarely transverse and descending colon.

Further invasion of submucosa and underlying blood

vessels facilitates migration of trophozoites to liver,
lungs and other organs from secondary lesions.
› Incubation period → 1-4 months Clinical Features
Asymptomatic disease
› 90% of the infections remain asymptomatic or mildly
symptomatic.
Symptomatic disease
› Diarrhea
– Gradual onset of lower abdominal pain and mild diarrhea
– Malaise, weight loss and diffuse lower abdominal pain
– Occasionally progressing to dysentery. › Amoebic dysentery
– Onset is gradual
– Colicky abdominal pain
– Fever
– Prostation
– Bloody diarrhea with mucus
and pus cells.
Resembles bacillary dysentery, a few clinical and laboratory features help to differentiate
between two diseases..
Amoebic appendicitis
– Acute right lower abdominal pain that mimics appendicitis.
Amoeboma
– May present as a palpable abdominal mass → confused with carcinoma.
Fulminant colitis or toxic megacolon
– Intense colicky pain
– Fever
– Profuse bloody diarrhea
– Mortalitiy rate is very high (40%) in patients with
corticosteroid therapy
– Note:- so important to exclude amoebiasis in patients
with inflammatory bowel disease.
Chronic non-dysenteric colitis
– Resembles inflammatory bowel disease with
potentially severe consequences if corticosteroids
are administered.
 Extra-intestinal Amoebiasis
› Commonest secondary lesion → liver abscess
→ postero-superior surface of the right lobe of
the liver.
› The pus of is made up of
– Liquified necrotic liver tissue and blood (thick
chocolate brown)
– Contains few leukocytes
– Bacteriologically sterile
› Trophozoite carried via the blood stream →
cause metastatic abscesses anywhere in the
body
Amoebic liver abscess (ALA)
Symptoms
– fever
– Abdominal pain localised to
the right upper quadrant
– Weight loss
– Pain may radiate to the right
shoulder and my be accompanied by a non-productive cough.
– On, examination→ tenderness of the liver, hepatomegaly,
jaundince less common.
Extra-intestinal Amoebiasis › Hepatic involvement  most common extra-intestinal complications of amoebiasis  reported in
2-10% of the individuals infected with E. histolytica. S. No. Clinical Features Amoebic liver abscess (ALA) Pyogenic abscess 1
Age 20-40 >50 2 Male:female ratio 10:1 1:1 3 Number of abscesses Solitary abscess in 80% of chronic ALA cases and about
50% of acute ALA Multiple abscesses in approx 50% of all cases 4 Diabetes mellitus Less common, approximately 2% of
patients More common, approx one fourth of the patients 5 Jaundice Uncommon Common 6 Pruritus Uncommon Common
Elevated bilirubin Uncommon Common Elevated liver enzymes Uncommon Common
› Lung
Pulmonary amoebiasis
– involved by extension of liver abscess through
diaphragm or
– by direct hematogenous spread from the intestine.
– Hepato-bronchial fistula → results with
expectoration of chocolate brown sputum → amoeba
can be demonstrated.
› Cutaneous amoebiasis
› Other organ involvement – Abscesses may develop
in (through hematogenous route)
› Brain (fatal)
› Spleen
› Kidney
› Genitourinary organ
› adrenals
 E. histolytica. Laboratory Diagnosis
• Stool examination:
Intestinal amebiasis has to be differentiated from bacillary dysentery.
• The stool should be collected into a wide mouth container and examined without
delay
• It should be inspected macroscopically as well as microscopically.
• Macroscopic appearance:
• The stool is foul-smelling, copious, semiliquid, brownish-black in color and
intermingled with blood and mucus.
• It does not adhere to the container.
• Microscopic appearance:
• Saline preparation: The cellular exudate is scanty and consists of only the nuclear
masses (pyknotic bodies) of a few pus cells, epithelial cells and macrophages.
 E. histolytica. Laboratory Diagnosis

• For the demonstration of cysts or dead trophozoites, stained preparations may be required
for the study of the nuclear character.
• Iodine-stained preparation is commonly employed for this purpose. Nucleus is clearly visible
with central karyosome
• Cysts shows a smooth and hyaline appearance Nucleus is clearly seen and no more than 4
nuclei are present
• The trophozoite of E. histolytica stains yellow to light brown.
• Routinely not used
 Diagnosis of Amebiasisof Amebiasis
RBCs are in clumps and yellow or brown-
red in color.
Charcot-Leyden crystals are often present
→ diamond-shaped, clear and Charcot-
Leyden crystals are often present →
diamond-shaped, clear and refractile
crystals.
Actively motile trophozoites throwing
pseudopodia→ Presence of ingested RBCs
Charcot-Leyden crystals
clinches the identity of E. histolytica.
Nucleus → a faint outline may be
detected.
 Entamoeba histolytica
› Cytoplasm→ visible with a clear, thin ectoplasm on the outside and a
granular “ground glass” endoplasm inside.
› Iron hematoxylin stain → clear visualization of nucleus → single, spherical
with a compact karyosome at its centre.
› Inner layer of nuclear membrane → lined with evenly distributed fine
chromatin granules.
› Food vacuoles with ingested RBCs and bacteria can often be seen.
Diagnosis of Intestinal Amoebiasis
› Microscopy
Media used for stool culture
 Boeck and Drbohlav media
 NIH (National institurte of health) polygenic
media
 Craig’s medium
 Nelson’s medium
 Robinson’s medium
 Balamuth’s medium and
diamond’s medium
 E. histolytica. Laboratory Diagnosis
Mucosal Scrapings
 Mucosal scrapings can be obtained by sigmoidoscopy useful in atypical presentations and
may serve as adjunct to conventional examination for Ova and cyst
 Direct wet mount, a permanently stained smear and immuno stained smears are examined.
 E. histolytica. Laboratory Diagnosis
Serological tests
• Indirect fluorescent antibody (IFA),
• Indirect hemagglutination assay (IHA),
Serum with antibody titer of 1:256 or more by IHA and 1:200 by IFA are considered to be
significant.
• Enzyme linked immunosorbent assay (ELISA), Serine rich E. histolytica protein (SREHP) are
detected using monoclonal antibodies by ELISA.
• Test for antibodies in serum help in diagnosis of mainly extraintestinal infections
• Counter-current immunoelectrophoresis (CIEP) and
• Latex agglutination tests.
Amebic antigen detection: patients with active infections and disappears after clinical cure.
Antigen
Lipophosphoglycan (LPG) amebic lectin
 E. histolytica. Laboratory Diagnosis

Molecular diagnosis
• Recently, deoxyribonucleic acid (DNA) probes and radioimmunoassay
have been used to detect E. histolytica in stool.
• It is a rapid and specific method.
• Real-time polymerase chain reaction (RTPCR) is a sensitive test for
detection E. histolytica from pus of liver abscess.
 Diagnosis of Amoebic Liver Abscess (ALA)
The pus in liver asabscess appear as red Anchovy sauce like appearance
The material aspirated is likely to to contain Trophozoites and may be
detectedand by direct microscopic examination
› Microscopy
– Trophozoites and cysts
– Direct microscopy of aspirated pus and sputum (Pulmonary rupture)

Stool examination: E. histolytica cyst can be detected in stool in

less than 15% cases of amebic hepatitis.
 Diagnosis of Extraintestinal Amebiasis
Liver biopsy
• Trophozoite of E. histolytica may be demonstrated in liver biopsy specimen, in case
of hepatic amebiasis or amebic hepatitis.
• Serological test, are of immense value in the diagnosis of hepatitis amebiasis
• Culture
• – NIH (National institurteu of health) media, Boeck and Drbohlav egg serum medium
containing Locke’s solution, Balamuth’s medium and diamond’s medium
• › Ag detection:- ELISA and immunochromatographic tests
• › DNA detection: PCR and real-time PCR.
• › Other diagnostic parameters:
• – Leukocytosis,
• –↑ Alkaline phosphatase
• – Radiology →ultrasonography
 Diagnosis of Extraintestinal Amebiasis
Serological tests:
• Indirect hemagglutination (IHA), highly sensitive, they often give false-positive
results.
• Latex agglutination (LA)
• Gel diffusion precipitation (GDP), less sensitive, but more specific.
• Cellulose acetate membrane precipitation (CAP) test
• Counter-current immunoelectrophoresis (CIE)
• Enzyme linked immunosorbent assay (ELISA)
 ELISAs are both sensitive and specific, become negative within 6 months of
successful treatment.
Serological tests remain positive for several years ever successful treatment
 A group of parasites with the same isoenzymes.

Zymodeme/isoenzyme analysis Zymodeme

Lectin binding Zymodeme analysis, genome

specific DNA analysis and staining with Monoclonal
antibodies have been successfully used as markers
to identify invasive strains of E.histolytica

Types of Zymodemes
Based on Electrophoretic mobility
E.histolytica strains are classified into
22 Zymodemes
However only 9 are invasive
 Emerging methods in Diagnosis

These are considered the most useful tests for detecting E. histolytica They test directly
for the parasite itself by exposing some stool to a strip of paper coated with antibodies.
The parasites will stick to the antibodies on the paper.
The test distinguishes E. histolytica from other parasites.
 Classification of Antiamoebics
• I. Tissue Amoebicide
• A. Intestinal as well as extra intestinal amoebicide
• i) Nitroimidazole derivatigves: Metronidazole, Ornidazole, Secnidazole(single dosage) ,
Tinidazole
• ii) Alkaloid: Emetine and Dehydroemetine
• B. Extra intestinal/hepatic amoebicide
• Chloroquine -in amoebic liver abscess is 1 g for 2 days followed by 5 g daily for 3 weeks.
• II. Luminal Amoebicide
• a. Amide Derivatives: Diloxanide Furoate
• b. 8-Hyroxyquinolines: Iodoquinol, clioquinol
c. Antibiotic : Tetracycline, Pramomycin
act in the intestinal lumen but not in tissues.
 Entamoeba histolyticaTreatment

Amoebic abscess is treated similarly to dysentery, with antibiotics. Sometimes surgical

drainage may be performed, but this is usually to rule out other usually ( (bacterial)
causes of abscess. It is also performed if an abscess is about to, or has ruptured

1- Metronidazole (flagyl) (3 times 3 mg/kg) every day (5-10) days .

• It kills the trophozoites and less effective against the cyst
• Most effective against the invasive amebae
• Less effective against the luminal amebae SO it is usually administered with a
luminal amebicide, such as iodoquinol or paromomycin
Alternatives to metronidazole include tinidazole, ornidazole, and nitazoaxanide
• 2- emetin hydrochloride. Diodoquine.
• 3- Dehydrometin - dihydrochloride. After that Titracyclen Cycle (5 days).
• 4- Dilodohydroxy quine (650 mg 3times, 20 days). Diloxanide furoate can also be
used in children >2 yr of age.
 Vaccines
Vaccines are being developed and tested for the treatment of Amebiasis.
The vaccine is a modified of the proteins expressed on the surface of E.
histolytica.
A study in rodentsна грызунах found that the vaccine prevented the
formation of liver abscesses, but much more research is needed to
determine if these vaccines are useful and safe in humans

https://www.slideshare.net/suprakashdas1/e-histolytica-
249981247
 Preventing Amoebiasis
Drink only bottled or boiled (for 1 minute) water, or carbonated
(bubbly) drinks in cans or bottles.
Fountain drinks and any drinks with ice cubes not safe. Water can
be made safe by filtering it through an "absolute 1 micron or less
" filter and dissolving iodine tablets in the filtered water.
Avoid fresh fruit or vegetables that were peeled by someone
else.
Avoid milk, cheese, or dairy products that may not been
pasteurized
NON-PATHOGENIC AMOEBAE
• Entamoeba dispar
• Entamoeba hartmanni
• Entamoeba coli
• Entamoeba gingivalis
• Endolimax nana
• Iodamoeba beutschlii
ENTAMOEBA DISPAR
• Non-invasive,nonpathogenic
• Earlier it was considered as a nonpathogenic strain of E.histolytica
• E.histolytica &E.dispar are morphologically identical
• Cysts of E.histolytica & E.dispar cannot be differentiated microscopically
ENTAMOEBA HARTMANNI
• Earlier considered as small race of E.histolytica
• Morphologically similar to E.histolytica
• But trophozoites &cysts are smaller and have a diameter of 4-12µm
&5-10µm respectively
• Trophozoites never contain ingested red blood cells
• LIFE CYCLE-Similar to E.histolytica
• Δ is by measurement of size of the trophozoites and cysts & absence
of red blood cells in trophozoites
 ENTAMOEBA COLI
• World wide parasite
• Nonpathogenic
• Habitat-Lumen of large intestine of man
• It exists in 3 stages-Trophozoite,Precyst and Cyst
• Life cycle-Similar to E.histolytica
TROPHOZOITES
• Sluggish movement
• Cytoplasm is not differentiated into ectoplasm and
endoplasm
• They never contain red blood cells
• But bacteria and cellular debris is present
• Karyosome is eccentric
• Nuclear membrane is thick and is lined by coarse
chromatin granules

CYST
Spherical,15-20µm
Nuclei-1to8
Chromidial bars are filamentous
PRECYST
Resembles in shape with that of E.histolytica
 ENTAMOEBA GINGIVALIS

• First parasitic amoebae to be recognised

• Commensal in the gingival tissue around
the teeth
• Described by Gros in 1849 in the soft
tartar between the teeth
• Also found in the diseased tonsils and in
the vaginal & cervical smears from
women using intrauterine devices
 ENTAMOEBA GINGIVALIS
• Only the trophozoite stage has been found ( no cyst stage)
• Encystation probably does not occur
• Infective stage TROPHOZOITE: 10-25µm in diameter;
• Diagnostic stage TROPHOZOITE
• Reproduction : asexually , binary fossion
• Actively motile by multiple pseudopodia
• Cytoplasm: Differentiated into clear ectoplasm and granular endoplasm
• Food vacuoles consists of digested leukocytes and epithelial cells
• Red blood cells are seen very rarely
• Nuleus: spherical; central karyosome
• Chromatin granules are closely packed
• TRANSMISSION: By close contact like kissing and from contaminated drinking utensils
ENDOLIMAX NANA
• World wide,nonpathogenic,small amoeba
• Habitat:Lumen of large intestine of humans,
primates and pigs
• It has 3 stages:Trophozoite,precyst and cyst
• TROPHOZOITES are small in size and cytoplasm is
demarcated into ectoplasm and endoplasm
• Motility: Sluggish
• Cytoplasmic inclusions:Bacteria, small vegetable cells and crystals
• No red blood cells
• NUCLEUS: Large irregular karyosome, arranged
eccentrically with Achromatic strands
• Chromidial bars and glycogen vacuole are absent
 IODAMOEBA BUETSCHLII
Nonpathogenic luminal parasite of large intestine
• Reservoir host : pigs
• TROPHOZOITES: Active in freshly
evacuated unformed stools &
sluggish in older stools
• Ectoplasm is not well differentiated
from endoplasm
• NULEUS relatively large; karyosome
surrounded by refractile globules
• Cytoplasmic inclusions: Bacteria and
yeast cells
• CYST:Uninucleate, Chromidial bars
absent.
 NAEGLERIA FOWLERI
Brain-eating amoeba Brain-eating amoeba

MORPHOLOGY
• 2 stages: Motile trophozoites and non-motile cysts
TROPHOZOITE:
• 2 forms→Amoeboid & Flagellate
• AMOEBOID: Amoebostomes- Distinctive phagocytic structures
• No peripheral chromatin. Reproduction is by simple binary
fission
• FLAGELLATE: Non-dividing, non-feeding form
• CYST: Uninucleate; spherical; mucoid-plugged pores or ostioles
in cyst wall
• Amoeboid form is the invasive stage
• INFECTION: Nasal contamination
during swimming or inhalation of dust
containing infective forms
• Flagellate and cyst forms could also
enter the nose
• Flagellate forms revert to amoeboid
forms and the amoeboid forms escape
from the cysts in the nose
 ACANTHAMOEBA Sp
A microscopic, free-living amoeba that
can cause rare, but severe infections of
the eye, skin, and central nervous system.
Several species of Acanthamoeba,
including A. culbertsoni, A. polyphaga, A.
castellanii, A. astronyxis, A. hatchetti, A.
rhysodes, A. divionensis, A. lugdunensis,
and A. lenticulata are implicated in
human disease.
The important species is A.culbertsoni

LIFE CYCLE STAGES Free-living trophozoites and

cysts occur in both the soil and freshwater.
ACANTHAMOEBA
Acanthamoeba spp. have been found in:
• soil ; fresh, brackish, and sea water
• heating, ventilating, and air conditioning systems
• mammalian cell cultures
• Sewage
• Vegetables
• swimming pools, medicinal pools
• human nostrils and throats ,
• human and animal brain,
• skin, and lung tissues.
•contact lens equipment; dental treatment units
• • dialysis machines
 MORPHOLOGY
• Acanthamoeba exists as active
trophozoites and resistant cysts
• No flagellate form
• TROPHOZOITES:24-56µm;l arger than
that of Naegleria;
• Irregular in appearance due to
acanthopodia- tapering like
pseudopodia;Motility-slow
• CYST:Double walled and so are quite
resistant
• Double wall-Outer wrinkled ectocyst
and inner endocyst
The trophozoites replicate by mitosis.
 Mode of infection
A. Granulomatous Amoebic B. Acanthamoeba Keratitis
Encephalitis •Through corneal trauma
•Nose to Lower respiratory tract •Exposure to contaminated
to Blood to Brain water
•Ulcerated skin and mucosa to •Wearing contaminated contact
Blood to Brain lenses
Symptoms • Symptoms
• Mental status changes body • Eye pain.
• Loss of coordination • Eye redness.
• Double vision • Blurred vision.
• Fever • Sensitivity to light.
• Sensitivity to light • Sensation of something
• Muscular weakness or in the eye.
• Other neurologic partial paralysis problems • Excessive tearing.
affecting one side of the
.
 When Acanthamoeba spp. enters the eye it can cause severe keratitis in
otherwise healthy individuals, particularly contact lens users .
 When it enters the respiratory system or through the skin, it can invade the
central nervous system by hematogenous dissemination causing
granulomatous amebic encephalitis (GAE) or disseminated disease, or skin
lesions in individuals with compromised immune systems
 ACANTHAMOEBA

• Pathogenicity and Clinical Features:

• Granulomatous Amebic Encephalitis (GAE) and disseminated infection
primarily affect people with compromised immune systems.
• Commonly seen in immunocompromised patients, including those with
neoplasia, systemic lupus erythematosus, human immunodeficiency virus
and tuberculosis
• It releases proteases→cytolysis of corneal epithelium
Incubation period:
• Unknown but estimated at weeks to months. The route of infection is
aerosol or direct inoculation with hematogenous spread to the CNS.
• Diagnosis:
GAE:-Demonstration of Trophozoites in CSF or trophozoites and
cysts in brain tissue
Keratitis:Trophozoites and cysts in corneal scrapings(wet mount
preparation)
Acanthamoeba Keratitis
Prevention and Control
• Store reusable lenses in the proper
storage case.
• 1. Storage cases should be rubbed and
rinsed with sterile contact lens solution
(never use tap water), emptied, and left
open to dry after each use.
• 2. Replace storage cases at least once
every three months.
Wash hands with soap and water and dry
before handling contact lenses.
ACANTHAMOEBA Treatment:
Granulomatous Amebic Encephalitis (GAE) Total excision of the mass and
treatment with pentamidine, usually in combination with one or more of the
following:
• Ketoconazole Acanthamoeba keratitis
• Hydroxystilbamidine - Therapy should include the cationic
• Paromomycin antiseptic agents, of which
• 5-fluorocytosine polymyxin chlorhexidine or polyhexamethylene
• Sulfadiazine biguanide (PHMB) is the most
• Trimethoprim-sulfamethoxazole effective. Dibromopropamidine+
• Azithromycin Propamidine isethionate ointment or
drops
- Ocular lesions –
- Enucleation of ulcer and corneal
transplan
 Balamuthia Mandrillaris
B. mandrillaris, a leptomixid free-living amoeba, is a newly
identified species reported to cause GAE.

It exists in amoeboid trophozoite stage. The flagellate stage is absent.

It is relatively large (12–60 µm), irregular in shape, and actively motile by broad pseudopodia.
Cyst of B. mandrillaris are usually spherical (6–20 µm),surrounded by a three-layered cyst wall—
outer irregular ectocyst, a middle mesocyst and an inner endocyst round wall. Under light
microscopy, it appears to have two walls—an outer irregular wall and an inner smooth wall.
 Infection is trasmitted through respiratory tract skin lesions; or eyes.
It causes granulomatous amoebic encephalitis in both healthy and immunocompromised hosts
particularly in children and elderly
 Life cycle is similar to that of Acanthamoeba spp.
 Key points of Amoebae
• E. histolytica is found in human colon and is mainly asymptomatic.
• Cyst contains glycogen mass and 1–4 chromatid bars.
• Pathogenic strains are identified by genetic markers and zymodeme analysis.
• Stools: In amoebic dysentery, stool are copious foul smelling, brownish black often with blood-streaked
mucus.
• Amoebic ulcers: Typical ulcers are discrete, flask-shaped, with ragged undermined margin, found in caecum
andsigmoido-rectal region
• Amoebic granuloma or amoeboma may develop from chronic ulcers.
• Extraintestinal complications: Amoebic hepatitis and liver abscess are the most common
• Abscesses in other organs such as lung, brain, spleen, and genitourinary tract may result from
hematogenous spreaor by direct spread from hepatic lesion.
• Diagnosis: By demonstration of trophozoites and cyst in stool and also by serological tests and imaging
techniques in hepatic amoebiasis.
Treatment: By Metronidazole or tinidazole along with parmomycin, diloxanide furoate, or chloroquine.
E. hartmanni, E. coli, E. gingivalis, E. nana, and Iodamoeba are commensals and non-pathogenic amoebae.
Naegleria and Acanthamoeba are pathogenic free-living amoeba.
N. fowleri occurs in 3 forms – cyst, trophozoite, and flagellate. It causes PAM.
Acanthamoeba species cause amoebic keratitis and also GAE in immuono-compromised subjects.