f'

[niicn iaurncl af Aninc! S:ience:; 69 i?.t

: iiS-90. February l999
Sfficiene;r ci three methods of oocyte recovery in the
ci rcrneelarl' camel
iCamelus
dromedoriws)
C N i'ilROHITi. IvIAHESH DAIT: and S S SHARMA3
isiesihan- Agriculture Llniversitl;, Bikaner, Rajasthan 334 001
FLccrjved: 27 Februa-1' 1998; Accepted: 26 August 1998
.{BSTRACT
(-)vari,:s
col!tc..c<j
jr+rrr ,.riarl fertale camels of Bikaner and surrounding areas during the breeding season were usecl for
{roc:icc..il,iriion.Fr:-rnitite leftorei 1Ifollicleswerecolleciedcomparedtoi9folliclesfromrightovary'.Ontheovarian
surfae e ifl ot ;ire 62 ibliicles i., ere haling erther blood ciots or haemorrhagrc margins. Follicles were well prcjecting on the
c..,3ilan si,.t'ace *:i lilir c-,ciallmean diameter * as 9.63t3.50 mrn. 36i 5b recovered oocytes were not having any curnulus
mass:ltr-arit:ij rc it. Arnr.'rist ih: 3 irieihods used for oocyte collection, aspiration was most efficient (32 ooc-\tes y'ielded
framu\oyaiies) cr.;lr':il:edtcf,rliicle di:,section(i-loccytesfrom5ovaries)orslicing(l0oocytesfromT
ovaries).Slicing
was con-.rd*rerl i+ b,e non-suitable irr ccilectioa of camel ooc]'tes because of the presence of haemorrhagic folliclss.
Key words: Camel. Follicles, Oocyte
in.;irr', l'li'tilizatiln is sttil in the experiinental stage in the
camelid.ie. l,in:bivcs rver*
pr"Cu:ed iit v'itt'c irorn llama ooc;vtes,
hcr\y.ivei'. ilter are stili no reilort?d llve +ffspring from
e:nbr-yos proci,:ceC ii," vttt,s, T'!re foilieular acti'/it)' is a
contiiluous ptuctsss i.n il;,: aniieal r-lilring the breeding season
anei when nct n;aied. ii3 i6'ilicle:; become attretic. rvhile,
continuousi-y olh.,::' foliicies matuie and gii,rw even in ihe
presence cfair aetive ccrpus luteutii (llravo 1994. Tibary and
Aiouas-si i996
).
Bou etc;/. (199j) i:r China described srnall-
scale studies o;t invltra ienilizalicn in rhe Arabian camel on
ovaries trom :laugi-:iereC aniin;,is. v.heieas Dei Campo er
ai.(1994a) d*scr.ii,ed lhe same in iiamas. in','ivrt collection of
oocytes far inviiit ierliiizl:iion-*'as described b1'Brogliatti el
al.{1996}. Tiirary ar:'i ,1-rr{)i.ia:r;! ( i t97). An aftempt ri'as made
to coliect the camel o-.'alies from recenill clied animals and
asses the e fficiency- of iiifferent medrcds cf ooclte retrieval,
MATERIAI-S AND TVIETHODS
'
Ovarie-s were cr,llected from dead f-emale dromedaries
witiiin 6 to 121rr *f-death from Bikaner ruea and its sunoundings.
The cornplete gt'nitalia *'ere ccllected wasi:ed and brought to
the laborar.;ry in werm nor:nal sallne. fort;fred ri ith antibiotics.
The ovaries $ ere sepaiatecl rn the laboratcry and the number
of follicles present on the respeciive cvaries. their position
and diameters were tLoted down. The oo.\'tes were collected
Present address:
jr
Assrstant I-'ioiesir,'r.
r
Professor,
Departmenl crf Obstetril.r att,j G)'naecology. College of
Veterinarv and Animal Science.
using TCM-199 fortified with antibiotics and bovine serum
albumin, by aspiration, follicular dissection and slicing
methods used for cattle (Katska and Smorag 1 984) arid buffalo
(Das er al. 1996). The morphology of oocy'tes was observed
under inverted phase contrast micrcscope. Only good sized
follicles (>8mm diameter) were used for follicular aspiration
and dissection.
RESULTS AND DISCUSSION
Follicles, 28 out of 62 that were seen on the ovaries were
having either blood clots or haemorrhagic margins (Fig'1).
This peculiar feature posed a great difficulry in collection and
assessment of the oocytes. In the absence of mating or
ovulation inciucing treatment, the fate of the maiure follicle
follows one of the two possible courses; atresia or cystic
degeneration (Tibary and Anouassi i996). The incidence of
anovulatory follicles in non-bred dromedaries may range from
35 to 50 % (Skidmore et al.1995). On the basis ofpalpabie and
Table l. E,fficiency of three methods of oocyte recovery in camel
Ovaries/ooc1'tes Method of collection Total follicle
present
Aspiration Slicing Puncture
-r.*----J"r.t
I r6r^r
ovary ovary
No. of ovaries taken 8
Recovery ofoocytes 32
7643
1,9
10 14
I
st
I
OOCYTE RECOVERY IN THE DROMEDARY CAMEL 89 February 19991
Figs 1-3. 1 Camel ovaries with haemorrhagic follicles and
mii.rgins. 2. Photo-micrograph shorving camel oocy'tes *'ithout
cumulus mass. 3. Photo-micrograph shorving camel oocl'tes rl ith
cumulus mass.
ultrasono graphic characteri stics, Tibary and Anouassi (1 997
)
defined 5 categories of anolulatory follicles in camelids, which
included thick-walled strucfure with blood clot and fibrin
sftands within the cavity (haemorrhagic foilicle). This explains
that the follicles with blood clots or haemorrhagic margins
observed during this study were anovulatory regressing
follicles, however, it is not known for certain why some of the
follicles develop into these large anolulatory,&aemonhagic
follicles and others do not.
The overall mean diameter of the follicles on the ovarian
surface was 9.63*3.50 (range 2-16 mm) ttre mean diameters are
lower than those reported by Homeida et al. (1988) by rectal
palpation and by Tibary et al. (1996) by ultrasonography in
the dromedary camels, moreover, the upper and lower limits of
the follicular diameters found during this sfudy were very low
compared to reports of Homeida et a/.( 1988) andTibary et al.
(1996), this could be due to difference in the method of
assessment of follicular diametrics and the fact that the ovaries
were collected from dead camels whose cause of death was
not investigated during this study. Skidm ore et al. ( 1 996) have,
however, reported the follicular diameter of 5-9 mm in dromedary
camels. The follicles were well projecting on the ovarian surface
except 3 thick follicles which were noted to be deepiy embed-
ded, and, not taken into account during this study. The recov-
ered oocy'tes (6a%; 35156) were denuded and did not have
any cumulus mass attached to it (Fig. 2). The shrunken nature
of the cytoplasm and the dark inner cellular mass in Fig. 3 (and
a common finding) could be because, although, due care had
been taken in collecting the genitalia from dead camels in time.
but the time lost from death of the camels to intimation for
collection was beyond control and probably the oocyes rvere
dead by that time. Del Campo et al. (1994b) reported that 17-
52oh ofthe oocytes are degenerated after recovery from the
follicles. By careful aspiration 4.0 oocytes per ovary (3214)
could be recovered whereas by follicular dis;ection only 2.33
(i4l6) oocytes could be recovered and by slicing only 0.7
ooc)'tes could be recovered per ovary (Table 1). 3licing method
was all the more unsuitable because plenty of blood rvas
extravasated into the medium of collection by the haemonhagic
follicles. Primarily follicular aspiration can be considered as
the most suitable method for retrieval of camel oocytes,
however, more sfudies need to be done. Methods of oocye
retrieval have been sparsely reported in camelids except the
in vivo methods like transvaginal ultrasounil guided methods
and hence comparison of the present study was not possible.
Horvever, our results are at variance with those of Del Campo
et al. (1994 a) who reported an average recovery of6.4 oocytes
per llama with the slicing method.
REFERENCES
Bravo P W. 1994. Reproductive endocrinology of liamas and
alpacas. Veterinary Clinics of North America. Food Animal
Practice l0 265-79.
Das G K. Jain G C. Solanki V S and Tripathi V N. i996. Efficiencl'
of various collection method for oocyte retrieval in buffalo.
TheriogenologY
,16(8):
1403-l 1.
Del Campo M R, Del Campo C H, Donso M X and Berland M.
1994a. In t;itro fertilization of llama (Llama glama) follicular
oocytes. Theriogenology 4l: 18'7.
Del Campo M R. Del Campo C H, Donoso M X, Berland l\{ and
Mapletoft R J. 1994b.1n vitro fertilizatron and development of
llamas
(Llanta glama) oocytes using epididymal spermatozoa
and oviduct cell-culture. Zlr eriogenology 4l:1219- 29 .
Homeida AM. Khalil M.R and Taha A A M.1988. Plasma
90
PUROHIT ETII.
[Vol.
69, No. 2
concentrations of progesterone. estrogens, testosterone and
LH like activity during the estrus c1'cle of the camel
(Camelus dromedarius). Journal of Reproduction and Fertilit.v
83:593-98.
Katska L and Smorag Z. 1984. The possibilities of recovering
cattle ooc), tes. LI oz l iw o s c i L z
"v
s kiw ani a O oc.r t ov B
"v"
d I e c.," c h
M e d1,cy na LI/e tery naryi na 40(8) : 499-5 02.
Skidmore J A, Billah M and Allen W R. 1995, The ovarian follicular
wave pattern in the mated and non-mated dromedary camel
(Canrelus dromedarius). Journal of Reproduction and Fertility
(Suppl) 49:545-48.
Skidmore J A, Billah M and Allen W R. 1996. The ovarian follicular
wave pattern and induction of ovulation in the mated and non-
mated one-humped camel
(.Came lus dr ome dar ius).Journal of
Reproduction and Fertility 106: I 85-92.
Tibary A and Anouassi A. 1996. Ultrasonographic changes of the
reproductive tract in the female camel (Camelus dromedarius)
during the follicular phase and pregnancy.Journal of Camel
P ractice and Re se arch 3 : 7 l-90.
Tibary A and Anouassi A. 1997. Theriogenology in Camelidae.
pp.l75-79. Abu Dhabi Printing and Publishing Company. Mina
Abu Dhabi. United Arab Emirates.