Oocyte Recovery Camel

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[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

seru

albumin,

by

aspiration,

follicular

dissection

and

slici

methods

used

for cattle

(Katska

and

Smorag

1 984)

arid buffa

(Das

er

al.

1996).

The morphology

of

oocy'tes

was observ

under

inverted

phase

contrast

micrcscope.

Only

good

siz

follicles

(>8mm

diameter) were

used

for

follicular

aspirati

and dissection.

RESULTS

AND

DISCUSSION

Follicles,

28

out

of 62

that

were

seen

on

the ovaries

we

having

either blood

clots or

haemorrhagic

margins

(Fig'

This

peculiar

feature

posed

a

great

difficulry

in collection

a

assessment

of the oocytes.

In the

absence

of

mating

ovulation

inciucing treatment,

the fate

of the maiure

follic

follows

one of the

two

possible

courses;

atresia

or cys

degeneration

(Tibary

and

Anouassi

i996).

The incidence

anovulatory

follicles

in

non-bred

dromedaries

may range

fro

35

to

50

%

(Skidmore

et

al.1995).

On the basis

ofpalpabie

a

Table

l.

E,fficiency

of three

methods of oocyte

recovery in

cam

Ovaries/ooc1'tes

Method

of collection

Total folli

presen

Aspiration

Slicing

Puncture

-r.*----J"

r

ovary

ov

No.

of ovaries

taken

8

Recovery ofoocytes

32

7643

10

14



st

I

OOCYTE

RECOVERY IN THE DROMEDARY

CAMEL

89

ebruary

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 ar

lower than those reported by Homeida

et

al.

(1988)

by recta

palpation

and

by Tibary

et

al.

(1996)

by

ultrasonography in

the

dromedary

camels,

moreover,

the

upper

and

lower limits

o

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 o

assessment of follicular diametrics

and the

fact

that the

ovarie

were

collected

from

dead camels

whose cause

of

death

wa

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

surfac

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 hav

any cumulus

mass attached

to it

(Fig.

2). The shrunken natur

of the cytoplasm

and the

dark

inner cellular

mass in

Fig.

3

(an

a

common

finding)

could

be because, although,

due care

ha

been taken

in

collecting

the

genitalia

from

dead camels

in

time

but

the

time

lost from death of

the camels

to

intimation fo

collection

was

beyond

control

and

probably

the oocyes

rver

dead

by that

time.

Del Campo et

al.

(1994b)

reported that

17

52oh

ofthe oocytes

are

degenerated

after

recovery from th

follicles.

By careful aspiration

4.0 oocytes

per

ovary

(3214

could

be

recovered

whereas by

follicular dis;ection

only 2.3

(i4l6)

oocytes could

be recovered

and by slicing

only 0.

ooc)'tes

could

be recovered

per

ovary

(Table

1).

3licing

metho

was

all

the more unsuitable

because

plenty

of blood

rva

extravasated into the

medium of collection

by the

haemonhagi

follicles.

Primarily follicular

aspiration can

be

considered a

the

most suitable

method for retrieval

of

camel oocytes

however, more sfudies

need to be done.

Methods of oocy

retrieval have been sparsely

reported

in

camelids

except

th

in vivo methods

like transvaginal ultrasounil

guided

method

and

hence comparison of the

present

study

was not

possible

Horvever, our results

are

at

variance

with

those

of

Del

Camp

et al.

(1994

a)

who reported an

average

recovery

of6.4

oocyte

per

llama

with

the

slicing method.

REFERENCES

Bravo

P W.

1994.

Reproductive endocrinology

of liamas an

alpacas.

Veterinary

Clinics

of

North

America. Food Anima

Practice

l0

265-79.

Das G K. Jain G C. Solanki

V S and Tripathi

V

N.

i996. Efficienc

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)

follicula

oocytes.

Theriogenology

4l: 18'7.

Del

Campo

M R. Del Campo C

H,

Donoso

M X,

Berland

l\{

an

Mapletoft

R

J.

1994b.1n

vitro

fertilizatron and development

llamas

(Llanta

glama)

oocytes

using epididymal

spermatozo

and oviduct

cell-culture.

Zlr eriogenology

4l:1219- 29

.

Homeida

AM.

Khalil

M.R

and Taha

A A M.1988.

Plasm



90

PUROHIT

ETII.

[Vol.

69, No.

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

follicu

wave

pattern

and

induction of ovulation

in the

mated and no

mated one-humped

camel

(.Came

lus dr ome

dar

ius).Journal

Reproduction and

Fertility

106:

I

85-92.

Tibary A

and

Anouassi

A. 1996. Ultrasonographic

changes of

t

reproductive tract

in

the

female camel

(Camelus

dromedariu

during

the

follicular

phase

and

pregnancy.Journal of

Cam

P ractice and

Re

se arch

3 : 7 l-90.

Tibary A and

Anouassi

A. 1997.

Theriogenology

in Camelida

pp.l75-79.

Abu

Dhabi

Printing

and

Publishing

Company.

Mi

Abu

Dhabi. United

Arab Emirates.

Oocyte Recovery Camel

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