What is the suitable media (easily prepared media) needed for cultivation and proliferation? what is the suitable method for cultivation this parasite?
#Entamoeba_Histolytica
Please see the following review paper for information and references:
http://cmr.asm.org/content/15/3/329.full.pdf+html
hi see this attach it is helpful
histolytica was first introduced by Boeck and Drbohlav in 1925
in a diphasic egg slant medium, and a modification of this
medium (Locke-egg) is still used today (35). Different
monophasic media that were developed for E. histolytica are
the egg yolk infusion medium of Balamuth (12), Jones’s medium
(96), and TYSGM-9 (41). Of the different media developed
for the xenic cultivation of E. histolytica, only three media,
diphasic Locke-egg, Robinson’s medium (150), and the
monophasic TYSGM-9 (41), are in common use (for details,
see reference 35).
Axenic cultivation involves the cultivation of parasites in the
absence of any other metabolizing cells (35). The axenic cultivation
of E. histolytica was first achieved by Diamond in 1961
(39). The monophasic medium TP-S-1 was developed and used
widely for culture of E. histolytica in different research laboratories
(35, 40). Currently TYI-S-33 (45) and YI-S (44) are the
most widely used media for axenic cultivation of E. histolytica
(35).
Culture of E. histolytica can be performed from fecal specimens,
rectal biopsy specimens, or liver abscess aspirates. As
the liver abscess aspirates of ALA patients are usually sterile
(98% cases) (19), addition of a bacterium or a trypanosomatid
is necessary before inoculation of amebae into xenic culture
(35, 53, 204).
The success rate for culture of E. histolytica is between 50
and 70% in reference laboratories (35). As culture of E. histolytica
from clinical samples such as feces or liver abscesses
has a significant false-negative rate and is technically difficult,
it is not undertaken in a routine clinical laboratory.
Entamoeba dispar can be grown in xenic culture; however,
most isolates grow poorly in monoxenic culture, and the
growth of only a few strains has been reported to be viable in
TABLE 1. Characteristics of trophozoites and cysts of common intestinal Entamoeba speciesa
Characteristics E. histolytica/E. dispar/
E. moshkovskii E. hartmanni E. coli E. polecki
Size, nuclei, and motility
Trophozoites 15–20 m, 1 nucleus (difficult
to see in unstained
preparations), actively
motile with finger shaped
pseudopodia
8–10 m, 1 nucleus (usually
not seen in unstained
prepn), usually
unprogressive
20–25 m, 1 nucleus (often
visible in unstained prepn);
sluggish, short, and blunt
15–20 m, 1 nucleus
(occasionally seen on wet
prepn), sluggish
Cysts 10–15 m, mature cyst with 4
nuclei, immature cyst has 1
or 2 nuclei (nuclear
characters difficult to see
on wet prepn)
6–8 m, mature cyst with 4
or 2 nuclei, two nucleated
cysts very common
15–25 m, mature cyst has 8
nuclei, occasionally 16 or
more nuclei
10–15 m, mature cyst with
1 nucleus, rarely 2 or 4
nuclei
Other features
Trophozoites
Chromatin (stained) Chromatin peripheral, may
have beaded appearance
Nucleus may stain more darkly
than E. histolytica, chromatin
may appear as solid ring
rather than beaded
(trichrome)
Chromatin clumped and
unevenly arranged, appears
as solid ring with no beads
Chromatin finely granular,
chromatin may also be
clumped at one or both
edges of membrane
Karyosome
(stained)
Karyosome small, compact,
centrally located but may
be eccentric
Karyosome usually small and
compact; centrally located or
eccentric
Karyosome large, not compact,
may or may not be eccentric,
may be diffuse or darkly
stained
Karyosome small and
usually centrally located
Cytoplasm (stained) Cytoplasm is fine, granular,
may contain bacteria;
presence of RBCs
diagnostic for E. histolytica,
although some E. dispar
strains may very
occasionally contain RBCs
Cytoplasm finely granular, may
contain bacteria, no RBCs
Cytoplasm granular with
differentiation into
cytoplasm and endoplasm,
vacuolated; bacteria, yeast,
and other debris may be
present
Cytoplasm is finely granular,
may contain ingested
bacteria
Cysts
Chromatin (stained) Chromatin peripheral with
fine uniform granules,
evenly distributed
Chromatin granules evenly
distributed (nuclear
characteristics may be
difficult to see)
Chromatin coarsely granular,
may be clumped and
unevenly arranged
Chromatin finely granular
Karyosome is small, compact,
but occasionally eccentric
Karyosome large, eccentric,
occasionally centrally located
Cytoplasm (stained) May be present;
chromatoidal bodies
usually elongate with blunt,
rounded, smooth edges;
may be round or oval;
chromatin may be diffuse
or absent in mature cyst;
clumped chromatin mass
may be present in early
cysts
Usually present; chromatoidal
bodies usually elongate with
blunt, rounded, smooth
edges; may be round or
oval; chromatin may or may
not be present
May be present (less frequent
than in E. histolytica),
splinter shaped with rough
pointed ends, may be diffuse
or absent in mature cysts,
clumped mass occasionally
seen in mature cysts
Abundant chromatoidal
bodies with angular
pointed ends, thread-like
chromatoidal bodies may
also be present, half of
the cysts contain spherical
or ovoidal inclusion mass
a Data are from references 57, 73, and 188.
VOL. 20, 2007 LABORATORY DIAGNOSTIC TECHNIQUES FOR ENTAMOEBA SPECIES 515
axenic culture, suggesting that E. dispar may be less able than
E. histolytica to obtain nutrients in a particle-free medium (29,
103). The use of different media for the culture of E. dispar has
been investigated, and these studies indicate that YI-S may not
be a suitable medium for the culture of E. dispar (35, 103).
For E. moshkovskii strains, culture media employed include
TTY-SB-monophasic with the trypanosomatid, TP-S-1-GM
monophasic for the axenic culture of amebae (40), and the
TP-S-1-GM monophasic medium (42). Other media containing
bovine serum used for culture of E. moshkovskii include
axenic medium TYI-S-33 with 10% bovine serum at 24°C (45)
or xenic medium TYSGM-9 with 5% bovine serum at either
24°C or 37°C (41).
Culture of E. histolytica in a clinical diagnostic laboratory is
not feasible as a routine procedure and is less sensitive than
microscopy as a detection method (35). Parasite cultures are
difficult, expensive, and labor-intensive to maintain in the diagnostic
laboratory (35). Overgrowth of bacteria, fungi, or
other protozoans during culture is the main problem encountered,
and therefore culture is not recommended as a routine
diagnostic procedure for the detection of Entamoeba species
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