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Vol 6 No 1, p 1-13 BIOTECHNOLOGIES IN CAMELID REPRODUCTION: CURRENT STATUS AND FUTURE PROSPECTIVES
Vol 6 No 1, p 1-13
BIOTECHNOLOGIES IN CAMELID REPRODUCTION:
CURRENT STATUS AND FUTURE PROSPECTIVES
G.N.Purohit
Department of Veterinary Obstetrics and Gynaecology, College of Veterinary and Animal Science,
Rajasthan Agricultural University, Bikaner-334 001, INDIA
ABSTRACT
Artificial insemination (Al) and embryo transfer are the two biotechnologies that have been applied to reproduction
in camelidae. Although during recent years Al has been developed in dromedary and bactrian camels, as well as in South
American camelids, methods of semen collection and evaluation are yet to be standardised. Moreover, poor sperm motility,
inefficient techniques of semen cryopreservation, inadequate procedure for timing insemination and ovulation are a few
of the turning
and priorities
points which when resolved in future will make Al a common place in camelidae. Global semen transport
for trait propagation will then come up. Development of MOET is the outcome of research in new and old
world camelids during the last decade. Although embryo recovery rates have been improved from 1.55/donor to 6.5/
donor, superovulatory treatments are yet far from being perfect. High variability in embryo recovery rates are thought to
be due to failure of ovulation, or abnormal leutinisation, superovulatory treatment or poor fertility and breeding management.
Pregnancy rates with fresh embryo transfer are currently too low (8-50%) and cryopreservation
of embryos need to be
developed
fully. Induction of ovulation is the common reproduction management technique used in camelids. Other
reproductive biotechnologies are in the primary stage of development. ln vitro fertilisation of follicular oocytes has been
sparsely
reported with development of embryos upto B-16 cell stage using epididymal sperm. Limited availability of slaughter
house
ovaries is one constraint which can be resolved by collection of COC's by in vivo methods. Work on such a
technique is in progress and standard protocols are in the offing. New biotechniques that are likely to be developed in
camelid reproduction in the nearfuture are gender preselection, nucleus transfer and embryo cloning, genome analysis
and gene mapping, in vitro embryo production, gamete cryopreservation and gene transfer.
Key words: Biotechnology, camelid, embryo, insemination, oocyte, semen, superovulation
Reproductive biotechnologies that have
extent due to various limiting factors. Some two
been applied for camelids include artificial
decades ago when the camel racing turned into
insemination
(Al),
multiple ovulation
and embryo
a serious sport (Haydn Evans and Wernery,
transfer, in vitro fertilisation and controlled
1995) concentrated efforts to improve the speed
breeding.
In light of the various developments and
characteristics of the dromedary began. These
constraints in the above techniques it appears
and other factors led to the specif ic use of males
mandatory to analyse the current status of these
biotechnologies in camelidae. ln this review an
attempt has been made to analyse the current
and the need for artificial insemination. Gordon
(1997) expressed that research to develop an
Al system for use in the Arabian camels is
artificial insemination, multiple ovulation and
necessary to improve the productivity in terms
embryo transfer, in vitro fertilisation and controlled
of meat quality, milk, riding and other purposes.
breeding and biotechniques likely to be developed
in f uture like gender preselection, nucleus transfer
Serious efforts on improvement th,rough Al in
and embryo cloning, genome analysis and gene
dromedary camels of many of the developing
countries had been lacking on account of their
mapping and gene transfer in this species.
Artificial Insemination
Although the f irst calf f rom Al was born to
a bactrian camel in 1960's
(Elliot,
1961) and
sole use for draught purpose, however, interest
on breed conservation is now emanating. In the
bactrian camel the technique of breeding with
Al is a development of the past two decades
(Chen and Yuen 1984, Chen et al, 1985; Xu ef
reports date back to as early
as 1966
a/, 1985, 1987,1990
and Zhao et a|,1992, 1994,
(Fernandez-Bae,a 1 966; Fernandez-Baca and
Calderon, 1968) in new world camelids but Al in
1996). Al in camelids has been reviewed by
Chaudhary (1995) and Tibary and Anouassi
camelidae has not been developed to a greater
(1ee7).
SEND REPRINT REQUEST TO G.N.PUROHIT
i-l
Journal of Camel Practice and Research
June 1999 / 1
a
a
Methods of semen coilection and evaruation Anouassi (1992) summarised the work and Both electroejaculation (Musa
Methods of semen coilection and evaruation
Anouassi
(1992) summarised the work and
Both electroejaculation (Musa et al,
lggz
expressed that
for short term preservation of
and 1993) and artif icial
vagina (AV) (Khan and
dromedary and
bactrian semen ail extenders
Kohli, 1973; Singh, l g8g)
have been used for
containing lactose or
egg yolk are suitable
collecting semen in the camelids. collection
semen in camelidae presents various problems
of
although laiciphos
appeutlo
be
better (sieme ef
a/, 1990). The dilution recommended
is
1:1
and
due to
dribbling nature of the ejaculation and the
1:3 and semen should be diluted onlV after
animal's
be
n of semen using the
complete
liquefaction and cooled slowly to 4-5"c.
AV used
ry
e
5 t=,t-t'r?""J;""?, 'irLX:
Dilution in order
to obtain a known concentration
d ro m e d a
such as 50x1 06
per cent
per ml has been
Anouassi
et al, l ggb; Willmen
advocated by some authors
1992). skimmed mirk extenders
(Anouass i et al,
used for equine
semen were also found to be
satisfactory.
lf
semen is
to be used within a few minutes of
collection,
storage at 37"c or room
temperature
is recommended
(Anouassi et
al,
f SbZy.
For
longer preservation
in a liquid form (upto 4g
hours) semen shourd be
coored srowry to 4 io soc,
which can be accomplished
by placing the tube
containing extended semen In
a water
et al,
bath which
1996a; Sumar,
19gg) the inner liner of the
is
then kept in a refrigerator (Tibary and
AV having annular
constrictions. A few
Anouassi, 1
gg7).
modifications to this
technique has been recenily
The dilution
varies from 1:1 to
of semen for cryopreservation
reported (Lichtenwarner ef
ar,
1gg6a,b).
coilection
1:g
for
the llama (Graham et
of semen using electro-ejaculation have been
al, 1978) and from 1:2 to 1:3 in
the dromedary
described in the dromedary (Er-Manna ef
ar,1gg6;
and bactrian
camel (Chen et al,
lggO; Graham
Graham et al, 1g7B; Merkt et al,1gg0; Merilan ef
et al, 1978; Zhao et
al,
1994,1 gg6)
the extenders
used
for deep freezing of
semen have been
adapted
from other spectes
containing
S_7%
glycerol as
cryoprotectant (Che n et al, f ggO; Sun
et al, 1990; Musa
et al,
1992, 1gg3;
Sie me et al,
1990;
Zhao et al,
1991)
and the
semen
is
packaged
in 0.25, 0.5 and 4 ml plastic srraws
(Musa et
al,
1992;
Sieme et
al, 19gO;
Willmen ef
and calderon, 196g)
and Anouassi, 1gg7),
but not recommended (Tibary
al, 1993) pellets
(Graham
et
al,
197B) or
because of the poor quality
ampoules (Chen et al,
1990; Zhao et al,
1gg4,
of the ejaculate obtained
and the distress caused
1996)
or 1 .5 ml straws
(Chen et al, 1990; Zhao
to the animar. other ress
important
methods
of
et
al,
1994, 1996).Semen
has been cryo_
semen collection in camelids include rectal
preserved
using
instant
freezilg
emproying dry
ice and liquid nitrogen.
gesidls
many otf,"r,
techniques a
simple technique ol
treezing
semen
packaged in
0.25 or 0.5 ml straws nas been
recently described
(Tibary and Anouassi,
19g7)
wherein dilution is
compreted as soon as the
semen is riquefied, the
diruted semen is coored
semen coilection have been
described in detair by,
down to 5"c over one hour and
maintained at this
Tibary and Anouassi (1997).
temperature lor Z hours before
packaging in
straws. The straws are placed on a
,".[
Semen Preservation
Both short and
4 cm
above the liquid
nitrogen surface for 1o minutes
long
term
preservation
of
and then transferred
direcily into
riquid
nitrogen.
camelid semen have been described. Tibary and
A standard protocol
for cryopreservation
of
2lJunetggg
Journalof Camel practice and Research

ts-

-

f
f
camelid semen is yet to be developed although f urther attention semen has been as
camelid semen is yet to
be developed
although
f urther attention
semen has been
as rack of adequate methods for
reported
to be stored for 6
semen collection,
poor post
(Tibary and Anouassi,
ejaculation sperm
1997) to B years (Graham
motirity, rack of standard
et al, 1978)
without loss of
techniques
roi
rreezing
motilify.
No
ieport
of
semen and
long distance
difficulty in transcervical
transport of f rozen semen and
pipette
passage in
south
resurtant birth of carves
American camef
ids.
They
are avairabre. The
thawing
stressed the need
temperature for small
for further research
straws is 37"C for 30
to:
l.Standardise semen
seconds (Tibary and Arouassi
cof lection techniques;
, 1gg7) whereas
2.Define procedures for
ampoules and large straws are thawed
semen
handling and
at 40_55"C
evaluation; 3.Standardise semen
cryopreser_
rn a water bath, for 30 seconds to 1 minute.
vation
techniques; 4.Standardise insemination
procedures for
Insemination
optimal
pregnancy rates. Besides
the
above, work on matJ
Oestrus females are
behavioural signs of oestrus
difficult to detect and
ano ?emale gamete
physiology
is required. Whife
considering
Al in
should be combined
camels for
with
impro.ving
assessment of ovarran
their genetic qualit-y,
activity by rectal
it
'proi"ction
is
palpation and/or
Anouassi, 1gg7).
worth noting that serection
lor
ultrasonography
mirr
(Tibary and
could pose probtems due to
The semen
deposition site
the fact tnat camefs
are very
recommended is the
late maturing and the
uterus, just
cranial to the
males used would
become
very
old
internal
before their
cervical os using
darght"r,.
an insemination gun or
performance
pipette by
are available and as such
l;;;
re_ctal or vaginal
i"r,
technique
(Tibary
and
storage of
Anouassi,
semen by cryopreservation
should
1gg7). studies on minimar number of
be
a viabre sorution
spermato zoa
and therefore stress be attached
per insemination are
.required
to it (Gordon, 1gg7).
lacking arthough
for
the
bactrian
camer 400 mirion
spermatozoa are recommended
(Chen et al,
Multiple Ovulation and
1 9BS, 1 990; Zhao et al, 1 991
Embryo
1 gg4,
Transfer
,
1 996) while
for dromedary a dose of SO million
Use of multiple ovulatibn
,p"rr"t
and
embryo
ozoa
transfer
is suggested
has been attempted since the
(Anouass t et ar, 1g92). various
197O
(Novoa and
problems are i
cY
however, the
following
artific
he
importan
embryo tran
most
of
work of
insemination
in
Weip
rY
(1995) concluded that
th
when the ,fi
transfer was born alt
camel Af is to ensure
ovulates. The induced
tha
al
appeared from lsraef (
nature of ovulation
usually
great deaf of work with li
poslng asynchrony with
insemination, seems to
emanated from
have been
resolved by
inseminating
work at UAE, due to interest in
at
fixed
intervars after
ovuration induced by the use of
the racing sport. The
a recent one, the maxi
work on MOET is however
hormones
like hCG,
GnRH etc (Che n et
al,1985;
chaudhary,
1gg5;.Tibary
and Anouass i,'
1gg7)
increased dose of semen for
1990 in the dromedary
Coope r et al, 1ggo, 1 gg
insemination
(as
1992; McKinno n et
al,
1g
semen in camel has been known
to
possess -et a
and llama (Bourke et
GnRH
like ovulation inducing factor, Chen
al,
McKenzie,
1985) and
1gg3; McEnvoy
mating with a tealer after
et al, 1gg2; Gatica et
Al (Tibary
al, 1994; Del
and Anouassi,
Campo et al,
1995;
Coirea
1gg7). By the concomitant
et
al,
use of
1994, 1997).
ultrasonographic
Embryo transfer has atso L""n
evaluation, insemination
is
reported in the
recommended when the
alpaca
(Sumar and
follicle is 12 to 1B mm
Garcia, 1986)
in diameter in one
with variable resurts.
seven rive
study (Tibary
births'1ir,ree
and
Anouassi,
alpacas and four llam
1997)
s) have resulted from
and 15 to 30 mm in another (Musa et al,
embryo transfer studies
1 ee2).
so far. Techniqr", of
embryo transfer used
Tibary and Anouassi
in
the dromedary have been
(1997)
summarised
appf ied to the bact rian
camel 1f iOary anO
the problems with Al in camelidae which need
Anouassi, lgg7).
Journal of Camel practice and Research
June lggg/3
ll"
a
porcine FSH (Super-Ov) in a result of using over others on the Superovulation dole schedule
porcine FSH (Super-Ov) in a
result of using
over others on the
Superovulation
dole schedule
decreasing
in the camelid
superovulation
Attempts'of
in
dromedaries' The super-
embryo recovery
yielded
poor and
have
generally
inOuced on day B-11 o{ the.,hCG
species
*u,
ovulation
an ill-defined
retults
because of
was dole thrice
inconclusive
mating
treatment at oestiur
Both FSH and eCG
"no
on
day
1g-14 following 5000
oestrus cycle lGorOon, 1997)'
ar l|hourly intlivaf s
have been ur"Jtotsuper
rvulation' Due to a short
at the first mating'
A total
21
lU of
hCG
,of
oestrus cycle and induced
of the 7 donors
ill-defined
luteal phase,
were recovered out
embryos
been
nature of
ovulation, the lr teal phase
l9u"
The numbet
flushed non-surgically'
?l ,:1t"lt
(Co1r91 et
induced using progestagen
rmplants
was maximum
In
embryos
yielding betwe
1992a
and 1995) '
al, 1 994; BourkJ
et
"iZ-12
to equal
divided doses of
"l'
this group compared
qan
be obtained without
arOerovulation
400 mg of Fortoiropin or
o'
oih"r
treatment but the best
5!1nO ot FS[-P-' Of the
in FSH-P and Follotropin
progesterone
fs eacn
treated
when treated females have
results are obtained
"nitui.
of FSH-P group yielded 2
r
group) only
iemafe
(Tibary and Anouassi'
\
no follicular
temates in both the groups did
"ttu"t""t
embryos, rest att
of
inducing
a luteal phase
in
methods
1997). Other
Using.220
not yield
ry^l:]-P
with
a t
ale
followed by non-
"nv
"tnorvo'
a
period
of 5
include mating
hourly
intervais over
decreasing
12
daV 7 and initiation
ot the uteri
ggZ) found a mean ovulation
surgical flushin!
1n
days Correa et at(f
on day
9
followed by usual
of
f
'J+3'1 in
of superovutatlon
by laproscopy)
rate (observed
f'CC and non-surgical
tr"utt"nt with
higher
compared
mating,
the
response
was
(lsmail
et
al'
Itamas and
embryo r,".ou" ry 6-7 day?,later
for 3 days
or 500 lU of
eCG daily
to 500 lU of
with
this method were
1993). Embryo
156 *;;isi
tor 4days (rvulation
rate of
'Ltou"ty
eCG +
5'33 corpora luteal were
total of 21
1-.6i,' although a
mean
i.oto'7, respeciively)' A
1.5+0.5 and
g to 47"/"
reco rded.
were collected correspondin
embryos
by 750 lU
CL observed. Ovulation was induced
Superovulation with FSH
^^
-:+^
nn
of hCG on day 5'
units
of ^{
ing 20 to 30
Porctne
or -,a
over 6 daYs (two.injections
tocols similar to those used
SuPerovulation with eCG
eCGhasbeenSuccessfuIlyusedfor
duv
g 2 daYs befo.re and
1500-
in'
]
in
dosage
f rom
,up"tovulation
induction of
course of Progestagen
6000lUinthedromedary(Anouassia,ndAli,
1990, 1992;
Skidmore
al'
1994;
McKinnon
et
Lt
1990;
Cooper
in good superovulation
ggZ;Yagil and Creveld' 1990) 500-
"t,1990;
Musa et al,
f
rea et al' 1997) 750
tne tlama ind (Cor
;,',i: li:3'i:?";,Ili'{
2000 lU in
lU in the alpaca (Bourke ef
a/' 1995; Del Campo
due
to
poor
probably
embryos,
respectively)
generally given-as a single
et ai,1995). eCG is
bvine FSH in decreasing
*"n-Jg"ment'
of completion
breeding
before or on the day
dose one day
of
1-3 mg b'i'd' for 3-5 days
or f ixed doses-
J"yt progesterone
treatment' The
of a 5 to r S
treatment
a 10-15 day progesterong.
variable with different
following
follicular response was
yielded equally
also been reporfed to haue
withi15o0 ltJ of ecG
have
('4
+ 3'2
doses ot eCc
(McKinnon et al'
good
Po.nse
with 2000
lU' Apouassi and Ali'
and 5 .7 + S.i
ssi'
1997)' Small doses
1994;
may be due to vaiiable ovulation
1990),
which
a single
dose) followed
of
ovi
The follicular population
U of
eCG have resulted
and
individual u"iiution.
by an
studied so far in camelids and
has been poorty
)
an
'Yos recovered Per treated
due to the follicular population
in
hence ,"utont
however' the
female (Skidmore et al' 1992)
with limited
cannot oe
ascribed to. Vyas (1 998)
iit"fy
to
be due to
the effect of eCG
eCG in lndian
effects ur"
data reported
poor
response
of
than that of FSH'
Porcine FSH (1-3 mg bid) in
over 3' 5 or 7 days after a 10-
fwo
dromedaries treated with 3000
dromedari"s.
on O"y 7 of 100 mg lM daily injections
decreasing
doses
lU of eCG
have also shown
15 day
proggsterone treatment
in a mean ovulatory
of progesteron"
resulted
(McKinnon et al'
to sup"rouriJte
a dromedary
5'5,
however' no embryo could be
response of
reported a beneficial
998) have
1994). Vyas (1
Journal of Camel Practice and Research
4/June 1g99
,r*.
a2
1992a', 1994 and 1995a,b)' Superovulation bY immunisation i1-+ the cuff with 3-5 ml of DPBS.
1992a', 1994 and 1995a,b)'
Superovulation bY immunisation
i1-+
the cuff with 3-5 ml of DPBS. The routine f lushing
supernatant.
Embryo recovery Per
donor
The initial low
recovery of 1'55 embryos/
to maximise ovulation
mating
or insemination
(Ano*tti
and Ali
1990; Cooper et al' 1992;
et al,1 994; Skidmore et al,1 996; Vyas'
McKinnon
1 eeB).
in future.
Embryo
No
sP
ues used for bovine or
lY used for camelids'
other
o does not enter the
Since th
6 or 6.5 after ovulation hence
uterus until day
attempts
to
collect embryos from the uterus
this stage will result in a low recovery rate
before
Ali'
(Adam et al, 1gg2;
Anouassi and
1990;
Bourke et al, 1992a;
McKinnon et a1,1994; Weipz
ovulatorytreatment,poorfertiIityorhinderance
and Chapman, 1985).
Flushing may be done in
the sitting position
or by placing the animal in
inembryodevelopmentandtransport.ovu|ation
June lggg / 5
Journalof Camel Practice and Research

lii;J:"",Xiffi';'i"x;,;X': ,xltii:t tH;'H "l l'";^""'iff:U'H'[: iH:T'!:':""1:ii:"1i::;:

rile,.::TT,"J3:il:3iiJ:r*:lHli![i[iil*:.fiiltt"t*'***l';li#:T[:!:J,

j*:?"3HlFi,tl*;rrn:tTJ"tlililt',;"Ff?p[;!::flilry6ii:ri,';.ti:r"J;]

of such

trearments :::i';;:l;"'"i"jbo-'s

turn

ifil'

lM-:l'lj:sesterone

"{::,li*li''l**,m;:*,W*',H,tiiii

rhe

reported.pr"sn,llSv

rates lrom

:lri

and Anouassi, 1ee7) rnsprle

:;;;;r;;

."t"

roricres,mav not ovurate and

"+?"Jlii""'u"Jtl!'"1""""'""'u trearment used i:t^l"Y'r;r,

"un u.o "it!"t '1"

"l!ill-i"il;"lJ-'1!":_"!!'! *llU,:*m*:t[iH

*U:tt#:i:

":1,?

L:::T;lT";i,';Jii"lnr",ifjmtn:nl;

l",8iff u;;;;;;;,"1"" "," kn:y1to

be arrected '

i:r{*r{Tl1dilillilrdfi

:in"ii'r:L'l",.#n::],ii,",'=m*:"f,],li:*f

recipilnt, however. rh-e,sde

i

lf ilYfl ilil ?3,J',11?l*"",", ,n" ,

';:l#"ll:: ?;ffi

);;;;;ti;. with increased

H::r

,l;'J":";:l:i rlltit*:rfTi,'J:i'":?l';l:

natural

malrnl

***t"""*ml

U#*q#i$t1ffift:i';ffi

or transfer,

,r1*r,,,,,,,,","i";ixi.x?i"n"?'j;ili"l,I::;it"+3i,:jti,:i{*+*+**;l+i:r.,?ix

a Progesterone

1 997).

The time

this stage.

:",T["J:?:::::t,

'^o'd',:i""7:i!" l"ln

ntnii*'";:*eipz and chapman' i;,,ttt'tt3l "'0"1"""i"*

used ror, {re.ezine

bovine

rePorted to'b:J'f: li;?:

,,'""mimr#',;;fl**ir:tr"tt'11;ij#:"',',ffi

*

b,:1,d.i;l*q:t

**'=*'i,rydl'ffi

##*i*nirutrrq,,t",l'":{ru hii"*'

:ll1r*n*:irH:tr1[1il:'?:;!,iii

1"0"".""'"t'""' nutririon'

studied

etc remains poorlv

em

rransrer or

Y"l,'-:'?i:l

Tinson, 1 992;

Bp,h surs

T;,

M(

;:*::r"Ut;en

iiift

:i[tJ;

ifflrf*inff

gTr:Tt"Ji:l::il:

rhe th-awins,ani

done as per procedures us'u rr

:rgfi:iigi'1'l;"""1,"iffi'J&il3ry;i'ill

,^"li*Uru';h;y],",1;::lfiHfjJ'.%:i

itJ;:,:"$ :iiln+Hi:[g11"jj"+1{* j["J,'"':.'tfilj

oi

",nOryo f o

r

described in freezins camerro e

reported but the n;-;;;i;al

Journal of Camel Practice and Researcr

6/June tggg

low epidural anaesthesia, a vaginal probe (5 or which does not survive the relatively large
low epidural
anaesthesia, a vaginal probe (5 or
which does not survive the
relatively large size
with a aspiration
needle and pump is
7.5 mhz)
(Tibary and Anouassi' 1997)'
process
vaginal wall to aspirate the
Ireezing
introduced through
major problems
encountered with
The
A recovery rate of 40-50"/o was
of embryo transfer
follicular contents'
efficient development
these workers whereas a recovery
in
camelidae are superovulation'
technology
reporteO by
rate of 64%
*"'
in the llama (Brogliatti
ovuIationandbreedingmanagementand
'"ported
et
of oocytes
from abattoir
al,1996). Retrieval
of
embryos'
More
research
cryopreservation
u
bvarie
follicular dynamics
should
be
directed towards
et al (1993) in
et a/ (1992 and
drome
of follicular
nypothalamopituitary
kers using
slicing
f"g'lation
1994)
f ixed time Al'
cycles,
ovuiation
induction'
wave "nO
SA fraction V'
25 PM
metho
in vivo' effect of
embryos
*orpnophysiology of
gentamicin
sodium
pyruvate and 50 pg/ml
on:amelid embryos and
cold temperatures
an average
recovery^of 6'4
to simplify and
sulphate) reported
studies on
follicular population
per
Ilama' Purohit
oo.Vt".
tackle the existing
improve the proceduies and
TCM
et a/ (1999) with
199 (fortif ied with
limited data using
problems.
BSA) described an average
antibiotics and
and 0'7. (1017)
recovery ot +.0 (g214), 2'33(.1416)
ln vitro fertilisation
follicular aspiration'
oocytes
per ovary by
has been attempted with
tn vitrofertilisation
camelids' Embryos have been
limited success in
f rom llama follicular oocytes
proOuced
in
vitro
oi dromedary ovaries.f rom
was considered unsuitable
of blood into the medium of
t al' 1994)
ePididYmal
using
per
cent of the oocytes were
but no live
birt
d' In the
ing to the
dissection and slicing
dead animals. Slicing
due to extravasation
collection. Sixty four
no cumulus mass attached to it'
denuded anO frLO
dromedary
IVF a
8-16 cell
stage in
ed (Bou et
success rates are very low and
OocYte Maturation
a/, 1993). Current
Not much
work has been done on the
been obtained from ejaculated
no embryo
has
maturation of
follicular oocytes' Maturation of
Most of the techniques used are adopted
semen.
by incubation
COC's is generally accomplished
f rom work in other sPecles'
i
with high
SX CO,
tension
at 38.5"C ,nO"
culture medium supplemented
humidity in tissue
Methods of oocYte retrieval
serum, pyruvate and antibiotics'
with hormones,
Two metfrods of oocyte
reported in camelids' Th e in
retrieval have been
vivo methods using
cent llama oocytes
unde^r such
Sixty
two per
ll
after 36 hours
incubation reached metaphase
(Tibary
and
laproscopy
.1n:i":i'^,t,nnt)
coc's matured
of culture whereas for dromedary,
ovum
pick
up
transvaginal
ultrasound guided
similar conditions the maturation
under nearty
and the in vitro
(OPU)
(Brogliatti
et al, 1996)
47% (Bou et
a:,1993).
lt appears that
rate was
complexes
methods
using
collection of oocyte
maturation is required for
(Tibary and
iong.r
time for'oocyte
after
overiect6my or post mortem
to other species such as the
camelids co.put"d
house material (Del
Anouassi,
t OSZl,'slaughter
24 hours)'
cow (36 hours vs
;'.
and recoy"t.y f rom
Cr.po
et al, lsigz ano
t994)
(Purohit et
al'
1999)'
dead
f emale camels
the in vivo methods surgical recovery
Amongst
SPerm PreParation
tnitial attempts
of Bou et al(1993) and Del
from the ovaries through a
of
mature follicles
99i,
1994) have been marginally
Campo
et ai(1
described for dromedaries (Tibary
laparoscopy is
sperm'
the former
successful' Using epididymal
the recovery rates
1997). HOwever,
*ill
and Anouassi,
oocytes
sperm
workers insemlnated
low as the follicles
.
are
described to be very
in medium containing
caffeine and
capacitated
bleed easily'
Laproscopic
become f r,ugit" and
hours' the oocytes
BSA for
2
hours. After 6-7
follicutar as[iration
can be accomplished in the
but not yet reported' Th.e best
to TCM-1 99 containing
FCS and
were transferred
llama ano alpada
43% with
pyruvate' A fertilisation rate of
sodium
described is
in vivo oocyte
collection technique
to the B-16 cell stage in vitro
guided follicular
embryos
developing
transvaginal
ultrasound
latter workers washed sperm
was
reported. The
liiOary and Anouassi' 1997)' Under
aspiration
June 1 ege l7
Journalof Camel Practice and Research
(Anouassi and Ali, 199 Skidmore et al, 1996 1996). SimilarlY in the I doses of
(Anouassi and Ali, 199
Skidmore
et al, 1996
1996). SimilarlY in the I
doses of GnRH (0.5-1
oocytes were penetrated by sperms'
ln vitro culture of embrYos
ln the llama embryos have been co-cultured
with llama oviductal epithelial cells' Del Campo
effective in inducing ovulation 26-30 hours later.
eProductive biotechno-
embrYo
logies
like artificial i
to
or being
trinsfe
I are likelY
for
ion and
routinely
used
Better
dissemination of
a/, 1 995).
at the race track has enjoyed a
performance
in development of such reproductive
pivotal role
Controlled breeding
in the dromedary but
Hormonal manipulation
of reproduction for
biotechnologies, especially
receive global attention for
the effort should now
early puberty,
out
of
season breeding and
identification,
preservation and promotion of other
lnduction of
follicular activity has been reported
of high
in camelids with limited success. Photoperiodic
traits like
milk production, production
capacity' In
control of reproduction
has been reported
quality fleece and f ibre and draught
potential of
success (Agarwal and
d"u"loping
countries the dairy
recently with limited
by the use of
1998) . The hormonal treatments are
dromedaries should be improved
Khanna,
based on injections
of eCG (Elias, 1990; Arthur
Al f ro m f rozen semen of males of the elite herd
to make them more important, where they are
and Al-Rahim,
1 982; Dahir et al, 1990; Elias ef
their primary role in transport'
losing
al, 1985)
FSH (Agarwal et al, 1993) or GnRH
two
Besides
the above
imPortant
(Homeidaefal,1991)andrepresentsmallerdata
and usually not supported
for
use
(Tibary
and
Anouassi,1997).Theuseofprostaglandinsfor
of reproductive
cycles
is of
little
use
management
in camelids
and instead induction of ovulation is
the common reproduction management technique
genome analysis and genel mapping and gene
used in camelids. For ovulation induction GnRH,
lM or lV (Anouassi et al, 1994;
transfer.
0.5 mg to 1 mg,
1985; Elias, 1990) Buserelin, 15 to
Bono et at,
Gender Preselection
20 ptg (Coope r et al, 1990 ,
1992; McKinnon and
1993; Skidmore et al,
It
may
be
advantageous
at different
Tinson , 1gg2;
Musa et al,
separation of X
situation and can be achreved by
1gg2,.1996)
and hCG 2500-4000 lU, lM (Anouassi
and Y chromosome bearing spermatozoa from
and Ali,
1990; Anouassi et al, 1994; Chen and
fertilising
ovum or the sexing
Che n et al,1 985; Coo per et al, 1990;
semen
and use for
Yuen , 1gB4;
of
embryos by recombinant DNA (rDNA)
lsmail et al, 1993; Skidmore et al, 1992,
1992:
them techniques proven in
have been successf ully used in dromedary
technology. Among
1996)
other species
include flow cytometry for sperm
and
baciiian camels. The best response is
separation and embryo sexing using PCR' Work
obtained when the follicles have the required size
vitro production of
(10-22 mm) and the uterus has maximum tone
on
in
embryo sexing and
Journal of Camel Practice and Researcl
8/June tggg

more and more studies \ ogY of male and female

*r

camelid embryos is in progress at various polymorphism can be effectively determined by laboratories and
camelid embryos is in
progress
at various
polymorphism
can be effectively determined
by
laboratories and it is likely that sexed embrvos
this
technique.
However, the primer used can
would be available in near f uture.
greatly
affect the
polymorphism
generated
(sherief and Arhadrami, 1996). Further studies
Nucleus transfer and
embryo ctoning
are needed using a wide
range
of
RApD
markers
A clone is considered
to
be
a large
to establish the choice of primers most suitable
population of identical molecules or cells that
for a particular camer popuration. porymorphism
arise f rom a common ancestor
(Granner,
1996).
has also been studied
in the ilamas and arpacas,
The basic procedure involves the utilisation
of
using
llama a genomic
DNA
library
(Lang et al,
micromanipulation and cell fusion to transfer
1997). Chromosomal
homology
between
blastomere
of
multicellular
embryos
into
dromedary and humans for ten
segments
have
enucleated oocytes. The nucleus of blastomere
reprogrammed in such a way that new embryo
been recenily established (Abdo et al,19g7).
rs
develoos.
Gene transfer
Although
cloned embryos have been
It is the basis of
producing transgenic
produced in rabbits,
mice,
sheep, goat,
cattle
and
animals. The transgenics
carry
rDNA molecule
pigs, there are still considerable limitations.
in their genome that are introduced
by
intentional
Nucleus transfer using somatic celrs became
the
human intervention (wall, 1996). The transferred
focus of scientific and public attention in 1gg7
gene consists of a functional part
element, the promoter. Over the
and a regulatory
variety of approaches and animal
last ten years a
species have
been used to
produce transgenic
animals.
Transgenic animals are
being
developed
for
a
variety of purposes including, models for human
it to be a more efficient and faster
way
of
making
diseases or for use in
human drugs, value added
transgenic
foetuses for cell therapies, adult
agricultural livestock with
improved
food product
animals for
protein
production and
organs for
or disease resistance varieties, human
xenotransplantation.
organ
For
camelidae
the
donors and bioreactors
to produce materiars for
multiplication of embryos by nucleus transfer
industrial and biomedical
applications
(Ziomek,
appears to be the primary locus of attention.
1998). Such advanced techniques are
likely
to
be incorporated in f uture research on cameridae.
Genome analysis and
gene mapping
It can be concluded that before more and
Molecular genetics
has tremendously
more
biotechniques are (for camelidae)
developed
increased the
understanding
of the structure
it is important
function and organisation of genes. Genome
use of existing
to standardise and maximise the
reproductive biotechnologies
namely Al and embryo transfer.
(1e64) ,Stroies on
Abder-Rouf M and
1,,'::';:ii,io
=
reproduction in camels (Cametus
dromedairus).
Mating
technique and coilection of semen. Journar of Veterinari
cost are fundamentar to the DNA
diagnosis in
Science, UAE '1 :1 13-1 1 9.
animal breeding. The
application
of
DNA
Abdo G, Rettenberger
diagnosis is in erimination of genetic disorders.
G,
Stranzinger G (1997).
Zoo FSH
analysis with seven
human chromosome specific
For camelids DNA anarysis techniques have been
libraries detects conserved
regions between human ano
camel (Camelus dromedarius).
Journal of Animal
Breeding and Genetics 1 1a (5): 369_375.
Adams CL, Bourke
DA, Kyle CE, young p and
McEnvoy
TG
(1992). Ovulation
and embryo recovery in the Llama.
Proceedings of rst
Internationar camet conference,
Dubai, UAE 125-127.
Adams GP, Sumar J and Ginther oJ (1991). Form and function
Journal of Camel practice and Research
June lggg/9
andFSH in llamas' Eurog to PMSG Ovarian responses of the corpus ruteum in ilamas (Lrama
andFSH in llamas' Eurog
to PMSG
Ovarian responses
of the corpus ruteum in ilamas (Lrama gtama).Veterinary
i""u'i
omlrican Camelids:75-B 1'
Symposi um ""
Record 125:127-1138'
P and Adam CL
CE' McEnugV T9'-'hung
Bourke DA, Kyle
a-nd embryo transfer
993)' Effect of low dose
n"tipil"i
tynchionisation
Aoarwal SP, Rai AK and Khanna
\?^f
(1995a).
ovarian activity
during non-
rio genology 41 :1 7 1'
on
South o*ti'""t'iamerios' The
of FSH administration
in
*t"rt'
rnoian
Journal of Animal
breeding
"u'o'n''i'i["
Science 63:387-390'
Bourke DA, Adam CL' Kyle CE' Younq P and McEnvoy TG
to eCG in llamas
(1995b)'
Superovuiatory
'"'poni"t
season breeding in
4 4:255 -260'
SP and Khanna ND (1998)' Off
dnol
ogy
t ti o g
(Lt am a
Aqarwal
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n' "it\"in
camel by
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BL (1993)'
Bravo PW, Stabenfeldt GH' Fowler ME and Lasley
Animal
associated with
Ovarian
.patterns
Embryo transfer in camel
";;';;"crine
and alpacs' Journal
A and Ali A (1990)'
in
llamas
Anouassi
reproductivt
of the workshop
Proce"iingt
"u""t*"iities
(Camelus oi'meirius)'
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Adnani M and
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l-1:d
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Palasz AT and A!a.ms GP (1996)'
Ultrasound
of
Proce.edings
to
achieve f"gn"nty'
Brogliatti
GM,
and oocyte
i'
UAE
175-177 '
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to.tti't-" aspiration
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tr,tnl*gin"t
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Zhao XX and Huang
Chen
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improve
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Successful
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291-
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prrgn"nty
Record 127 '
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VeterinarY
a n d
: ":
;
c h e n s H H, B u r t D w
I
3 X :
? 1o^s:,"^1'
?,il
ffi
?"""ltt"?
3
i
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maplng'
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in PoultrY
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S;;
DA,
r6t"u'
Proceedinss Svmpostum
Bourke
embryo
transfer in
Pt;;;;ii;;'
Animal
pregnancy tollowing
1o,1,-t.Ytgl:al
Record cited
Veterinary
Series 1 :168-178'
Editor)'
tn"e
llamas tr-ettei'io
ay
Anouassi' 1997'
A
(1994)
bY TibarY and
Combarnous Y and Anouassi
as
immunosorben horm
""
and
enzYme linked
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ns
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"-t^I1"ifl.,;ffi'#:FJ"iJ
aids to ttlivi"g-piiuitutv
el'
came'
of the
ll:'f:l ;:'":'$i^T:"iii:1
i'n'i"in'"iionur
reproductiJJ't""n"g"ment
183-185'
Journal
oi
nlii
Environments 26: 1 5-20'
UAE
Conference' Dubai'
,'
Billah
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Cooper MJ,
e and
Bourke DA,
tl^. llama
(Llama
An
and ;i;;
wn'treeo)'
an aio to?iir,"orlJo
n,""d^ing
!n
AA
n the
as
and
Record 1 30 :424-428'
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sYnch'onit"
g I am a)'Veteri nary
ngs of
embrYo
camels for
dromedarY
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Journalof Camel Practice and Research
10/.lunetggg
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191 .
Corre
t t"'Ji
ational
2:7BB-
790.
I
corre
;:":i3i'"J"i:
I
nation' Animal
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Del-CamPo MR, Donoso MX, Del-C
C, Parrish JJ and MaPletoft RJ
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