protozoa

Protozoa

Single-celled eukaryotic microorganisms belonging to

kingdom protista are classiffed as Protozoa (Greek
Protos: first; zoon: animal).
Characteristics of Protozoa:
 Protozoan parasites consist of a single "cell-like
unit" which is morphologically and functionally
complete and can do all functions of life
They do not have cell wall; but they have outer
membrane , flexible layer, pellicle called plasmalemma
Plasmalemma:- It is a tri-layer membrane, consist of
protein with lipoprotein layer; plasmalemma has a
selective permeability property
 They are made up of a mass of protoplasm
differentiated into cytoplasm and nucleus
 The cytoplasm consists of an outer layer of hyaline
ectoplasm and an inner granular endoplasm. The
ectoplasm functions in protection, locomotion, and
ingestion of food, excretion, and respiration
 In the Endoplasm, there are different vacuoles
responsible for storage of food, digestion and
excretion of waste products. in addition to other
organelles such as Nucleus, Golgi bodies, and
mitochondria.

 The nucleus also functions in reproduction and

maintaining life. There are two type of nucleus:
1. Compact nucleus , or
2. Vesicular nucleus

 Some protozoa, have more than one nucleus such

as ciliates ,which have two different size nuclei ,and in
some flagellates there are two equal size nuclei.
 Protozoa reproduce by asexual means, (Binary
fission, Multiple fission, Budding) although in some
groups sexual modes.
Some grops used fission following conjugation for
reproduction like Balantidium coli.
 Protozoa have the ability during their entire life
cycle or part of it to move by locomotors organelles or
by a gliding mechanism.

Protozoa have heterotrophic mode of nutrition,

whereby the free-living forms ingest particulates,
such as bacteria, yeast and algae, while the parasitic
forms derive nutrients from the body fluids of their
hosts.
Classification of Protozoa:
•The classification of protozoa is mainly based on their
means of locomotion.
•They are subdivided into the following four classes :-
1. Sarcodina:
•Motility is due to the out flowing of ectoplasm,
producing protoplasmic projections called pseudopodia
(false feet). Examples: Free-living form like Negleria
fowleria and parasitic form like Entamoeba histolytica.

2. Mastigophora:
•Locomotion is effected by one or more whip-like, thin
structures called flagella. Examples: Free- living forms like
Euglena and parasitic forms like Trichomonas vaginalis,
Trypanosoma gambiense, Giardia lamblia.
3. Ciliophora:
•Locomotion is carried out by means of short hair-like
projections called cilia, Examples: Free-living forms
like Paramecium, and parasitic form like Balantidium
coli.

4. Sporozoa:
•Unlike the above three classless of protozoa,
members of the class sporozoa do not have
locomotor organelles in their mature stage; however,
immature forms exhibit some type of movement.
• All the members of this group are parasites.
Examples: Plasmodium, the malarial parasites of
animals and human beings.
Types of Protozoa:
Based on the mode of nutrition, protozoa are
of the following two types:
1. Free-living protozoa: They ingest
particulates, such as bacteria, yeast and algae.
2. Parasitic protozoa: They derive nutrients
from the body fluids of their hosts.

Based on the habitat , protozoa are of the

following three types:
1.Protozoa of alimentary canal .
2.Protozoa of body tissues and blood.
3.Protozoa of urogenital tract.
Sarcodina: Amoeba
 Amoebae are structurally simple protozoans which
have no fixed shape.
 The word “amoeba” is derived from the Greek
word “amoibe” meaning “change”.
 They change their shape due to presence of an
organ of locomotion called as “ pseudopodium”
 Amoebas primitive unicellular microorganisms
with a relatively simple life cycle which can be divided
into two stages:
• Trophozoite – actively motile feeding stage.

• Cyst – silent, resistant, infective stage.

 Reproduction occurs through binary fission,
and budding.
Cyst is formed in unfavorable conditions and is
usually the infective form for vertebrate host
(e.g. Entamoeba histolytica).

 Motility is accomplished by extension of

pseudopodia (“false foot”)
 A moebae are classified as intestinal amoebae
and free living amoebae. .
 The most important is the Entamoeba which
found in human involves three groups with
similar shape, and they are:
A- Histolytica group
B- Coli group
C- Gingivalis group
 There are many conditions exist in the
intestine affect the distribution of parasitic
amoebae, these are:
1- Degree of pH
2- Abundant of oxygen
3- Co2
4- Concentration of bile juice
Pathogenic Intestinal Amoeba
Entamoeba histolytica
 E. histolytica is worldwide in distribution but more
common in tropical and subtropical countries.
 E. histolytica has three sub species—E. histolytica
subspecies histolytica , dispar and moshkovskii ,
Cysts and trophozoites of all the three subspecies are
morphologically indistinguishable
 E. histolytica is the pathogenic species causing
amoebic dysentery and a wide range of other invasive
diseases, including amoebic liver abscess, where as
other two are considered as nonpathogen that
colonize the large intestine.
Morphology
E. histolytica has three stages:-
(1) tropho zoite,
(2) precyst
(3) cyst (immature and mature).
Trophozoite(troph.)
 It is the invasive form as well as the feeding and replicating
form .Some times the troph. found in the feces of patients with
active , acute disease.
 It measures 12–60 μm (average 15–20 μm) in diameter.
Cytoplasm of troph. is divided into a clear ectoplasm and a
granular endoplasm.
 Granular endoplasm contains red blood cells (RBCs), white
blood cells (WBCs) and food vacuoles containing tissue debris
and bacteria. RBCs are found only in the stage of invasion
Pseudopodia: Ectoplasm has long finger like projections called
as pseudopodia (organ of locomotion).
 Nucleus is single, spherical, 4–6 μm size, contains
central dot like compact karyosome surrounded by
a clear halo.
Nuclear membrane is thin and delicate and is lined
by a layer of fine chromatin granules. The number
of chromosomes varies between 30 and 50

 Amoebic trophozoites are anaerobic parasites. They lack

mitochondria, endoplasmic reticulum and Golgi apparatus.

Precyst
 It is the intermediate stage between trophozoite and
cyst.
 It is smaller to trophozoite but larger to cyst (10–20 μm)
 It is oval with a blunt pseudopodia. Food vacuoles and
RBCs disappear. Nuclear structures are same as that of
trophozoite
Cyst
It is the infective form as well as the diagnostic form of the parasite found
in the feces of carriers as well as patients with active disease.
It measures 10–20 μm (average 12–15 μm) in diameter.
Nuclear structures are same as in trophozoites.
First, the cyst is uninucleated; later the nucleus divides to form binucleated
and finally becomes quadri-nucleated cyst
Cytoplasm of uni-nucleated cyst contains 1–4 numbers refractile bars with
rounded ends called as chromatoid bodies (aggregation of ribosome) and a
large glycogen mass (stains brown with iodine)
Both chromatoid body and glycogen mass gradually disappear, and they
are not found in mature quadrinucleated cyst
Cysts are present only in the gut lumen; they never invade the intestinal
wall.
Minute” form of Entamoeba histolytica:
They are the commensal phase of E. histolytica, living in the
lumen of gut. They are usually smaller in size (trophozoite 12–14
μm and cyst < 10 μm) and often mistaken as E. hartmanni.
Trophozoite of Entamoeba histolytica

Hematoxylin stain Trichrome stain

saline mount
Iodine mount
shows
shows
mature cyst
immature cyst
(with four
(three nuclei)
nuclei)
 Life Cycle
Host: E. histolytica completes its life cycle in single
host, i.e. man.
Infective form: Mature quadri-nucleated cyst is the
infective stage .
It can resist chlorination, gastric acidity and
desiccation and can survive in a moist environment
for several weeks.

Note: Trophozoites and immature cysts can be passed

in stool of amoebic patients, but they can’t serve as
infective form as they are disintegrated in the
environment or by gastric juice when ingested.
Mode of transmission :
1. Feco-oral route (most common): By ingestion
of contaminated food or water with mature
quadri-nucleated cysts.

2.Sexual contact: Rare, (especially in developed

countries among homo sexual males).

3.Vector: Very rarely, flies and cockroaches may

mechanically transmit the cysts from feces,
and contaminate food and water.
Development in man (small intestine)
1.Excystation: In small intestine, the
cyst wall gets lysed by trypsin and a
single tetra-nucleated (four-
nucleated) trophozoite (metacyst) is
liberated which eventually
undergoes a series of nuclear and
cytoplasmic divisions to produce
eight small metacystic trophozoites
(amoebulae).
Development in man (large intestine)
Metacystic trophozoites (amoebulae) are
carried to large intestine and multiply by
binary fission, and then colonize on the
mucosal surfaces and crypts of the large
intestine especially the cecal and
sigmoido-rectal regions, where they feed
on bacteria and cellular debris and
undergo repeated the cycle of binary
fission.
The parasite lives in the colon lumen causing no harm
(minute forms), or under certain conditions, trophozoite
invades the intestinal wall, and some times the
trophozoite ( magna forms), transports by the portal
blood to the liver and rarely to lungs, brain, & the skin
where they multiply, depending on different factors :-
host susceptibility,
age, sex,
nutritional status,
 host immunity,
intestinal motility,
Cyst are never found in the tissues of intestine, they are
fully lumen forms and are unable of invasion.
Encystation:
After few days, the trophozoites transform into
precysts then into quadri-nucleated cysts which are
liberated in feces
Encystation occurs only in the large gut.
 Factors that induce cyst formation include food
reduction , over-crowding i.e. aggregation of
trophozoites in the mucin layer, & accumulation of
waste products.
Mature quadri-nucleated cysts released in feces
can survive in the enviornment and become the
infective form.
Immature cysts & trophozoites are some times
excreted, but get disintegrated in the environment.
Pathogenesis :-
•Trophozoite of E. histolytica is the major
invasive form
•It possesses many virulence factors that
play role in the pathogenesis of intestinal
as well as extra-intestinal amoebiasis.
1-Amoebic lectin antigen
2-Amoebapore
3-Cysteine proteases
4-Hydrolytic enzymes
5-Neuraminidase and metallo-collagenase
Pathogenesis of Intestinal amoebiasis
Trophozoites invade the colonic mucosa
producing characteristic ulcerative lesions and
profuse bloody diarrhea (amoebic dysentery).
Males and females are affected equally with a
ratio of 1:1.
Amoebic ulcer :
•The classical ulcer is flask-shaped (broad base
with a narrow neck).
•It may be superficial or deep ulcer
• Size ranging from pin head to some inches .
Complications of intestinal amoebiasis (cross section of intestinal
wall)
 Bloody diarrhea
(amoebic dysentery).

fever

abdominal pain
nausea, anorexia
 Pathogenesis of Extraintestinal amoebiasis
Following 1–3 months of intestinal amoebiasis, about 5–10%
of patients develop extraintestinal amoebiasis.
Liver is the most common site (because of the carriage of
trophozoites through the portal vein) followed by lungs, brain,
genitourinary tract and spleen.

Amoebic liver abscess

•The most common group affected: Adult males (male and
female ratio is 9:1).
•The most common affected site is the posterior superior surface
of the right lobe of liver.
•Abscess is usually single or rarely multiple.
•Amoebic trophozoites occlude the hepatic venules; which leads
to necrosis of the hepatocytes.
•Inflammatory response surrounding the hepatocytes leads to
the formation of abscesses .
Anchovy sauce pus: Liver abscess pus is thick
chocolate brown in color. The fluid is acidic and
pH 5.2–6.7 and contain of necrotic hepatocytes

Cross section of liver

showing
Anchovy sauce pus amoebic liver abscess
(right side)
A . Laboratory diagnosis of Entamoeba histolytica; B. Laboratory diagnosis of
amoebic liver abscess.
Treatment
1. Metronidazole or tinidazole is the drug of choice
for intestinal amoebiasis and amoebic liver
abscess.
2. Other measures include fluid and electrolyte
replacement and symptomatic treatment.

Prevention
1. A voidance of the ingestion of food and water
contaminated with human feces.
2. Treatment of asymptomatic persons who pass
E. histolytica cysts in the stool may help to
reduce opportunities for disease transmission.
 Entamoeba hartmanni

cyst

Trophozoites

Non-Pathogenic
Amoeba
Trophozoites
cyst

cyst
Trophozoites
 Entamoeba coli
•E. coli was first described by Lewis (1870) and its
presence in healthy persons was reported by Grassi
(1878).
•It is worldwide in distribution
•E. coli is a nonpathogenic commensal amoeba that
colonizes the large intestine.
•It has also three forms—trophozoites, precyst and
cyst
Trophozoites :-
•larger than E. histolytica about 20–50 μm with
sluggish motility and contains ingested bacteria but no
red cells.
•The nucleus is clearly visible in unstained films
and has a large eccentric karyosome and thick
nuclear membrane lined with coarse granules of
chromatin
Cysts :-
•large, 10–30 μm in size, with a prominent glycogen
mass in the early stage.
•The chromatoid bodies are splinter like and irregular.
•The mature cyst has 8 nuclei
•The life cycle is the same as in E.histolytica except that
it remains a luminal commensal without tissue
invasion and is nonpathogenic.
 Eccentric karyosome
Irregular chromatins granules
Trophozoites

Cysts
Entamoeba coli

Cysts
Entamoeba hartmanni

E. hartmanni occurs wherever E. histolytica is found.

It is now considered to be a separate species of
nonpathogenic commensal intestinal amoeba.

It is much smaller than E. histolytica, the

trophozoite measuring 4–12 μm and cyst 5–10 μm in
size
•Trophozoites do not ingest red cells and their motility
is less vigorous.
• The cyst resembles that of Endolimax nana.
It is also known as small race variant of E.
histolytica, i.e. morphologically it is similar to E.
histolytica but of smaller size (trophozoite is 8–10
μm and cyst is 6–8 μm.
It is nonpathogenic and colonizes the large
intestine
 Its life cycle is similar to E. histolytica
Entamoeba gingivalis

It is the first parasitic amoeba of humans to be

described; recovered from the soft tartar between the
teeth.
It is unusual in two respects:
1. It inhabits in the mouth rather than in the large
intestine
2. Only trophozoite stage exists; no cystic stage
Trophozoite is similar to that of E. histolytica
trophozoite except
Smaller in size (10–15 μm)
The cytoplasm contains food vacuoles with ingested
bacteria, leuocytes, and epithelial cells.
Nucleus similar to that of E.
histolytica, nucleus is round with
central karyosome lined by
coarse chromatin granules.
The amoeba lives in gingival tissues and is
abundant in unhygienic mouths.
 It is a commensal and is not considered to cause
any disease
 It is transmitted by direct oral contact.
 E. gingivalis rarely found in bronchial washings
and vaginal and cervical smears, where it can be
mistaken for E. histolytica.
Endolimax nana

This common commensal amoeba is widely

distributed.
It lives in the human intestine.
The trophozoite is small (nana: small), less than 10
μm in size with a sluggish motility
The nucleus has obvious karyosome which is
eccentric and irregular and connected to nuclear
membrane by one or none coarse strands.
The cyst is small, oval, and tetra - nucleate with
glycogen mass and chromidial bars, which are not
clear or absent
It is non-pathogenic.
Iodamoeba buetschlii

This is widely distributed, and less common

than E. coli and E. nana.
The trophozoite is small, 6–12 μm, with
clear nucleus
The prominent karyosome is half the size of
the nucleus, having ox’s eye appearance.

The cyst is oval, uni-nucleate , and has a

prominent iodine staining glycogen mass
(iodophilic body). Hence, the name
‘Iodamoeba’.
It is non-pathogenic
Cyst
 Entomoeba coli trophozoite

trophozoite

Cyst
Cyst
Troph. Cyst

Entamoeba gingivalis

Troph.
No cyst