CLINICAL PARASITOLOGY Laboratory

CLASS LOBOSEA-AMOEBA
NEIL ALLEN ELQUINTO, RMT
COLLEGE OF MEDICAL TECHNOLOGY
CENTER FOR MEDICAL & ALLIED HEALTH SCIENCES
NEW ERA UNIVERSITY
 INTRODUCTION
• Protozoa is a unicellular parasites.
• These are most commonly found in the lower gastrointestinal tract.
• Most of species can form non-feeding, non-motile cysts,this stage of life
cycle could be infective stage in humans.
• This is also considered as the transfer stage of most species. Transmission
of amoeba is generally by ingestion of cysts in fecally contaminated food
and water.
• Nuclei of each species are quite different for the different genus of
amoeba. For this reason, identification of stained nuclear structure aids
in diagnosis.
• In cystic stage, the number of nucleus varies as well, which is also
crucial for correct identification. Other diagnostic features present in
the cystic stages includes size, shape of cyst and other inclusion bodies.
 INTRODUCTION

• The trophic stage (vegetative stage), diagnostic features includes size,

cytoplasmic inclusions and type of motility exhibited. The trophozoite is the
motile and feeding stage of amoebas.
• Trophozoites can be destroyed by iodine
• A specialized structure known as pseudopodia enables the organism to move.
• This structure arises from the cytoplasm and is characteristic structure for
amoebas.
• Entamoeba histolytica is the medically important species in this group and is
the cause of amoebic dysentery in humans. This organism can occur in other
primates, dogs, cats, and rats.
• All other organisms (amoeba) are considered as non-pathogenic.
 INTRODUCTION

• If the organism is genus Entamoeba (true amoeba) – peripheral chromatin

(visible nuclear membrane in both trophozoite and cyst) and chromatoidal in
cyst only.
• If the organism is genus Endolimax or Iodamoeba (other amoeba) – neither
peripheral chromatin or non chromatoidal body are present.
 Pathogenic Amoeba: Entamoeba histolytica
Non-Pathogenic Amoeba: Entamoeba dispar,
Entamoeba moshkovskii, Entamoeba hartmanni
Features of Entamoeba histolytica
Features of Entamoeba histolytica

Trophozoites
• It measures 12–60 μm (average 15–20 μm) in diameter
• Cytoplasm of trophozoite is divided into a clear
ectoplasm and a granular endoplasm
• Granular endoplasm looks as ground glass appearance
and 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 fingerlike
projections called as pseudopodia(organ of
locomotion); which exhibits active, unidirectional
rapid progressive and purposeful movement
 Features of Entamoeba histolytica
Trophozoites
• 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
• The space between the karyosome and the nuclear
membrane is traversed by spoke like radial
arrangement of achromatic fibrils (cart wheel
appearance)
• Amoebic trophozoites are anaerobic parasites. They
lack mitochondria, endoplasmic reticulum and Golgi
apparatus.
Features of Entamoeba histolytica
Cyst
• 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
quadrinucleated cyst
• Cytoplasm of uninucleated 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.
 Features of Entamoeba dispar
• E. dispar is morphologically indistinguishable(both
cyst and trophozoite) from E. histolytica, so it may
be considered as a subspecies of E.histolytica.
• It can be distinguished from E. histolytica by:
• Zymodeme study (hexokinase isoenzyme pattern)
• Molecular methods, PCR amplifying small subunit rRNA
gene)
• Detection of lectin antigen in stool
• RBC inside trophozoites—present only in E. histolytica.
• It was described by Brumpt in 1993
• It is nonpathogenic, usually colonizes in the large
intestine (10 times more than E. histolytica) but
doesn’t invade intestinal mucosa
• It grows well in polyxenic media, however, poorly
grows on axenic media
• E. dispar doesn’t induce antibody production.
Features of Entamoeba moshkovskii
• E. moshkovskii is also morphologically indistinguishable from E. histolytica
and E.dispar (may be the third subspecies of E. histolytica).
• Th is species was fi rst described from Moscow sewage by Tshalaia in 1941
and was thereafter reported to occur in many different countries including
India
• It can be distinguished from E. histolytica by isoenzyme analysis,
molecular methods and detection of lectin antigen
• Though it is a nonpathogen harboring inintestine but recent studies from
Bangladesh and India have reported E. moshkovskii as a sole potential
pathogen in patients presenting with gastrointestinal symptoms and/or
dysentery, highlighting the need for further study to investigate the
pathogenic potential of this organism.
Features of Entamoeba hartmanni
• 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.
Features of Entamoeba hartmanni
• 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.
 Non-Pathogenic Amoeba: Entamoeba coli;
Entamoeba gingivalis; Entamoeba poleckii,
Endolimax nana, Iodamoeba butschlii
Features of Entamoeba coli
• E. coli is a nonpathogenic amoeba that
colonizes the large intestine.
• The life cycle is similar to E. histolytica
• It has also three forms—trophozoites,
precyst and cyst
• It is frequently found in the stool
samples of healthy individuals and
should be differentiated from that of E.
histolytica
Features of Entamoeba coli
Features of Entamoeba gingivalis
• It is the first parasitic amoeba of Trophozoite is similar to that of E.
humans to be described; recovered histolytica trophozoite except:
from the soft tartar between the • Smaller in size (10–15 μm)
teeth.
• Larger food vacuoles containing WBCs
• It is unusual in two respects: (only Entamoeba species that contains
1. It inhabits in the mouth rather WBCs)
than in the large intestine
• Nucleus similar to that of E. histolytica
2. Only trophozoite stage exists;
no cystic stage
Features of Entamoeba poleckii
• It is a nonpathogenic amoeba • Cyst is of 5–11 μm size and has
usually found in the intestine of pigs one nucleus with features
and monkeys. similar to that of trophozoite.
• However, human infection is rare, • It has many chromatoid bodies
mainly restricted to Papua New with threadlike ends (like E.
Guinea where it is the most coli) and cytoplasm has a large
common intestinal amoeba in nonglycogen inclusion mass
humans
• The trophozoites measure 10–12 μm
size, motility nonprogressive and
sluggish (like E. coli) and contains
one nucleus having central
karyosome and fine peripheral
chromatin (like E. histolytica)
Features of Endolimax nana
• It is a small (nana means small) nonpathogenic
amoeba.
• It is worldwide in distribution, frequently
resides in the large intestine of humans and
other animals
• Trophozoite measures 8–10 μm in size and
shows sluggish motility
• Cyst is 6–8 μm in size and contains one to four
nuclei. Cytoplasm doesn’t have chromatoid
body or glycogen vacuole
• Nucleus (both trophozoite and cyst)—
Karyosome is eccentric and irregular;
fromwhich several achromatic strands extend
tothe nuclear membrane. There is no
peripheral chromatin on nuclear membrane
Features of Iodamoeba butschlii
• It is also worldwide in distribution though • Cyst measures 10–12 μm in size, round to
less common than E. coli and E. nana oval and mostly is uninucleated
• Trophozoite is 12–15 μm in size. The • Nucleus has central karyosome surrounded by
refractile chromatin granules (bull’s eye
ectoplasm and endoplasm are not appearance or basket nucleus). On
differentiated. permanent smear, the nucleus may appear to
• Cytoplasm is more vacuolated. Nucleus is have a halo surrounding the karyosome
similar to that of the cyst • Cytoplasm of the cyst contains large iodine
stained glycogen mass or iodophilicbody
(hence named as Iodamoeba) and no
chromatoid body
 GUIDE QUESTIONS: Please cite your
references using the APA format
• Describe and illustrate the life cycle of Entamoeba histolytica
• Discuss the pathogenesis and laboratory diagnosis of Entamoeba histolytica ? And what
serological properties are present in Entamoeba histolytica compared to the rest of
commensals entamoeba
• List the free-living amoeba. Write in detail about their life cycle, pathogenesis and
laboratory diagnosis ?
Differentiate between:
(a) Features of Entamoeba dispar and Entamoeba histolytica
(b) Amoebic dysentery and bacillary dysentery
(c) Features of Entamoeba coli and Entamoeba histolytica
(d) Features of Naegleria and Acanthamoeba
(e) Features of Acanthamoeba and Balamuthia