tehnica LOPU

8/13/2019 tehnica LOPU

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OOCYTE RECOVERY FROM LIVE GOATS

Oocytes from live goats can be retrieve follo!ing s"#erov"lation or !it$o"t $ormonal

stim"lation "sing vario"s #roce"res% T$ese incl"e follicle as#iration or slicing of

ovariectomi&e ovary' follicle as#iration t$ro"g$ la#arotomy' LO() or T)GA%

La#arotomy or stanar s"rgery met$o of OR is invasive' fre*"ently ca"sing a$esion

of ovaries an ot$er viscera an ot$er s"rgery relate com#lications' even eat$ +Melican

an Gavin' ,--./% In t$eir e0#eriment' "sing s"#erov"late goats +Sama123 et al%' ,---/

retrieve 4.%- oocytes #er goat t$ro"g$ as#iration of follicles after ovariectomy an .%5

oocytes #er goat t$ro"g$ as#iration of e0teriori&e ovaries by la#arotomy% T$e LO()#roce"re is recogni&e as less tra"matic t$an t$e stanar la#arotomy +6oeman et al%'

,--57 Tibary et al%' ,--8/% Recently' T)GA tec$ni*"e $as been intro"ce in goat OR

!$ic$ is non9invasive in nat"re +Graft et al%' 4:::7 Melican an Gavin' ,--./%

Recoltarea ovocitelor e la animalele vii #rin te$nica la#aratomiei re#re o

meto; inva&iv;' #ro"ci alte com#lica=ii legate e interven=ia c$ir"rgical;' c$iar >i moartea animal"l"i

+Melican >i Gavin' ,--./% ?n e0#eriment"l lor' folosin ca#re s"#"se #rotocol"l"i e

#oliov"la=ie' Sama12 >i col%' ,---' a" re">it recoltarea a 4. ovocite@ca#r; #rin metoa

as#ira=iei folic"lilor "#; o#era=ia e ovariectomie >i .'5 ovocite@ca#r; "tili&i Gavin' ,--./%


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In T)GA tec$ni*"e' t$e goat is #lace in orsal rec"mbence an a 8 MB& transvaginal

trans"cer' attac$e to t$e "ltraso"n "nit' is #ositione vaginally for oocyte as#iration

+Graft et al%' 4:::7 Melican an Gavin' ,--./% T$is a##roac$ $as #rove to be a safe'

#ractical alternative to s"rgical or la#arosco#ic oocyte collection% Bo!ever' efficiency ofT)GA9erive oocytes for IV( is not !ell st"ie li1e LO() yet +Melican an Gavin'

,--./% T$e T)GA is commonly "se in large omestic livestoc1' #artic"larly cattle an

not m"c$ #ractice in small r"minants s"c$ as goats% T$is tec$ni*"e can be #roblematic

in t$e goat' "e to #remat"re l"teal regression' lo! fertili&ation rates an variability in t$e

s"#erov"latory res#onse to e0ogeno"s $ormones +Cogni23' 4:::/% In a st"y' n"mber of

follicles etecte an oocytes $arveste "sing T)GA +:%8 an %5' res#ectively/ !as less

t$an for goat obtaine by LO() +4D% an 4%' res#ectively/ +Graft et al%' 4:::/% T$e#ercentage of oocytes recovere from goats s"becte to t$e T)GA +./' $o!ever' !as

similar to t$ose s"becte to t$e LO() +:/% It $as been re#orte t$at re#eate T)GA

in #re#"bertal calf ca"se $istological lesions in t$e ovary !$ic$ may alter normal

ovarian f"nction an co"l #ossibly affect f"t"re fertility +Snel9Oliveira et al%' ,--,/%

T$is tec$ni*"e co"l offer an alternative' com#are !it$ traitional breeing' for

#rogeny evelo#ment from female transgenic fo"ner airy goats in near f"t"re +Melican

an Gavin' ,--./%

?n te$nica T)GA' animal"l este a>e&at #e masa e o#era=ie n #o&i=ie orsal;' iar

c" a"tor"l "n"i trans"ctor transvaginal e 8 MB& ata>at la "n ecograf se reali&ea&;

as#irarea ovocitelor +Graft >i col%' 4:::7 Melican >i Gavin' ,--./% Aceast; aborare s9a

oveit a fi o alternativ; sig"r; >i #ractic; la metoele c$ir"rgicale sa" la#arosco#ice e

colectare a ovocitelor% C" toate acestea eficien=a metoei T)GA n" este la fel e bine

st"iat; #rec"m LO() +Melican >i Gavin' ,--./% Te$nica T)GA este frecvent folosit; nefectivele e animale mari' n s#ecial la bovine% Metoa este #roblematic; la

r"meg;toarele mici' n s#ecial la ca#re' in ca"&a regresiei l"teale #remat"re' ratelor

re"se e fertili&are >i variabilit;=ii r;s#"ns"l"i #oliov"lator +Cognie' 4:::/% ?ntr9"n

st"i" efect"at e Graft >i col%' 4:::' n"m;r"l folic"lilor etecta=i >i al ovocitelor


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recoltate folosin te$nica T)GA +:'8 >i res#ectiv '5/ este mai mic eceterii n"m;r"l"i e escenen=i +Melican >i Gavin' ,--./%

LA(AROSCO(IC OV)M (IC69)( OR LO() I GOATS

T$e LO() is one of t$e best tec$ni*"es for oocyte recovery from live goats% T$e LO()follo!e by IV( $as been #ro#ose as an efficient met$o for t$e #ro#agation of s$ee#

an goat of $ig$ genetic val"e +Jalassarre et al%' 4::' ,--,7 Tervit' 4::7 Cogni23 et

al%' ,--/% T$e LO() is avantageo"s t$an stanar la#arotomy9base met$o !$ic$

allo!s re#etition of t$e la#arosco#ic #roce"re more fre*"ently an more times "ring

t$e re#ro"ctive life of a val"able female +Jalassarre et al%' ,--D/% Anot$er avantage of

LO() is t$e #ossibility of #ro"cing embryos an #rogeny from animal categories t$ose

are not able to re#ro"ce by m"lti#le ov"lation9embryo transfer +MOET/ an artificialinsemination +AI/' incl"ing #re#"bertal +Jalassarre an 6arat&as' ,--/ an age goats

+Jalassarre et al%' ,--D/% T$e LO() tec$ni*"e obviates several ca"ses of t$e #oor

res"lts observe !it$ s"#erov"lation' s"c$ as #oor ov"lation rate' early regression of

cor#"s l"te"m +CL/ an #oor fertili&ation +Jalassarre an 6arat&as' ,--/% T$erefore'

collection of oocytes after e0#os"re of t$e ovary by la#arotomy $as been gra"ally

re#lace by LO() tec$ni*"e in small r"minants%

La#arosco#ic ov"m #ic19"# re#re "na intre cele mai b"ne te$nici e

rec"#erare a ovocitelor e la animalele vii% Aceast; te$nic; "rmat; e IV( a fost #ro#"s;

ca o meto; eficient; e nm"l=ire a ovinelor >i ca#rinelor e mare valoare genetic;

+Jalassarre >i col%' 4::' ,--,7 Tervit' 4::7 Cognie >i col%' ,--/% Te$nica

la#arosco#ic; este mai avantaoas; ec


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re#etarea #roce"rii e mai m"lte ori n tim#"l vie=ii re#to"ctive a "nei femele e

valoare +Jalassarre et al%' ,--D/% )n alt avanta al te$nicii LO() l constit"ie

#osibilitatea e a ob=ine embrioni >i escenen=i e la iferite categorii e animale care

n" s"nt ca#abile #entr" re#or"c=ie #rin te$nica MOET +m"lti#le ov"lation9embryotransfer/ >i ns;m


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$ormonal treatments' may be re#eate at intervals as s$ort as a !ee1' even ays

+Alberio et al%' ,--,7 Gibbons et al%' ,--D/' !it$o"t iminis$ing goo *"ality oocytes

$arvest +Stangl et al%' 4:::7 Alberio et al%' ,--,/%

Bo!ever' re#eate LO() in "nstim"late goats also #rovie oocytes +9 #er oe #er

session/ +Cogni23 et al%' ,--5/% If t$e *"ality of t$ese oocytes for IV( is confirme' t$is

met$o co"l #rovie a !ay to #ro"ce offs#ring from genetically val"able females

!it$o"t "sing $ormones +Cogni23 et al%' ,--5/%

Rec"#erarea ovocitelor #rin intermei"l te$nicii LO() e la onatoare vii +ca#re

>i oi/ se reali&ea&; #rin #"nc=ia folic"l"l"i >i a#oi #rin as#irarea con=in"t"l"i folic"lar c"

"n ac asc"=it g$iat la#arosco#ic% (rim"l e0#eriment la ovine reali&at c" a"tor"l te$niciiLO()' efect"at n an"l 4:D e c;tre Snyer >i K"1elo!' a #ermis rec"#erarea a

ovocite in ,4 e folic"li% Treb"ie men=ionat c; #oten=ial"l te$nicii LO() s9a reali&at n

#aralel c" e&voltarea te$nologiilore IV( +Jalassarre >i col%' 4::' ,--,/% )lterior'

LO() evine o meto; eficient; e colectare a ovocitelor' ati

col' 4::7 $$ol&er 62 >i col%' 4::D/ ci la ca#rine +Jalassarre >i col%%' ,--,7

(ierson >i col%' ,--/%

?n veerea rec"#er;rii ovocitelor #rin intermei"l #roce"rii LO() treb"iene#lini=i "rm;torii #a>i

aneste&ia general; a animal"l"i7

conten=ia n ec"bit orsal a onatoarei7

as#irarea folic"lilor s"b control la#arosco#ic c" a"tor"l "n"i ac e 4.9,- G

montat la o #i#et; e #lastic' conectat; la "n t"b e colectare >i la o linie e vi%

Interven=ia "rea&; mai #"=in e 5- e min"te@animal >i e#ine e n"m;r"l e

folic"li as#ira=i +62 $$ol&er >i col%' 4::D/' #"ti col'

4:.7 Stangl >i col%' 4:::7 Alberio >i col%%' ,--,7 (ierson >i col%' ,--/%

?n ca&"l animalelor s"#"se "nor m"lti#le tratamente $ormonale re#etarea te$nicii

LO() se reali&ea&; la intervale mici e tim#' &ile +Alberio >i col' ,--,7 Gibbons >i col%'


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,--D/' f;r; afectarea calit;=ii ovocitelor recoltate +Stangl >i col%' 4:::7 Alberio >i col%'

,--,/% C" toate acestea' a#licarea LO() la oi nestim"late $ormonal a #ermis ob=inerea a

9 ovocite@sesi"ne +Cognie >i col%' ,--5/% ?n ca&"l n care calitatea acestor ovocite se

confirm;' aceast; meto; ar #"tea oferi o cale e a #ro"ce escenen=i e la onatoarevaloroase in #"nct e veere genetic' f;r; a "tili&a $ormoni +Cognie >i col%' ,--5/%


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In s"#erov"late goats' oocyte recovery rate range from 55 to .- !it$ an average

yiel t$at range bet!een 8% an 45% oocytes #er animal +62$$ol&er et al%' 4::D7

Alberio et al%' ,--,7 Jalassarre et al%' ,--,7 Jalassarre et al%' ,--57 Jalassarre an

6arat&as' ,--7 (ierson et al%' ,--7 Gibbons et al%' ,--D/% It $as been fo"n t$at LO()can be re#eate "# to 8 times in t$e goats at ifferent intervals an in ifferent seasons

!it$ little or no im#ortant c$ange in overall res#onse +(ierson et al%' ,--7 Ra$man et al%'

,--Da/% Li1e ot$er mammals' goat oocyte ac*"ires t$e ability to mat"re an accom#lis$

its evelo#mental com#etence "ring oogenesis an follic"logenesis% (revio"sly it $as

been s$o!n t$at t$e #ro#ortion of evelo#mentally com#etent oocytes increases !it$

follic"lar si&e +B"mblot et al%' ,--87 Ban et al%' ,--7 6$atir et al%' ,--D/% Conse*"ently'

t$e timing of ov"m #ic19"# affects oocyte evelo#mental com#etence% In $"mans+T$ornton et al%' 4::-7 Manso"r et al%' 4::/' mon1eys +g et al%' ,--,7 C$en et al%'

,--/ an #igs +Rat1y et al%' ,--5/ ov"m #ic19"# !as #erforme 5 $ after $"man

c$orionic gonarotro#$in +$CG/ inection% Maority of t$e oocytes collecte at t$is

interval are at t$e mat"re or meta#$ase II stage an are evelo#mentally an meiotically

com#etent% Bo!ever' in goats' oocytes collecte 5 $ after $CG inection are still at t$e

immat"re stages an are mat"re in vitro for ,D $ before reac$ing meiotic com#etence

+Jalassarre et al%' ,--57 Ra$man et al%' ,--Db' ,--./% 6ee#ing t$is in min o"r recentt$at elaying LO() #ost9FSB #l"s $CG treatments can im#rove oocyte retrieval rate an

oocyte *"ality +Ab"lla$ et al%' ,--./% T$erefore' LO() at - an D, $ #ost FSB #l"s

$CG treatment co"l be t$e #referre #rotocol to o#timi&e yiels of goo *"ality oocytes

for IVM an ICSI embryos in goats% As alreay mentione earlier' abattoir ovary so"rce

is very limite in Malaysia' t$erefore' LO() co"l be an alternative #roce"re for goat

OR to carry o"t IV(' es#ecially ICSI st"y% Timeline of significant fining of LO()

e0#eriments in goats is e#icte


8/22

?n ca#re s"#erov"late' rata e rec"#erare ovocit"l"i a variat e la 55 la .-' c" "n

ranament mei" care a variat ntre 8' >i 45% ovocitelor #e animal +62 $$ol&er et al'

4::D7 Alberio et al%' ,--,7 Jalassarre et al%%' ,--,7 Jalassarre et al%' ,--57 Jalassarre

>i 6arat&as' ,--7 (ierson et al%' ,--7 Gibbons et al%' ,--D/% Sa constatat c; LO() se

#oate re#eta e #i com#eten=a sa e e&voltare n tim#"l oogenesis >i follic"logenesis% Anterior' acesta a

fost emonstrat c; #ro#or=ia e evelo#mentally cre>te com#etent; ovocitelor c" o

imensi"ne a folic"lar +B"mblot et al%' ,--87 Ban et al%' ,--7 6$atir et al%' ,--D/% ?n

consecin=;' moment"l e ov"l"l"i #ic19"# afectea&; com#eten=a ovocit"l"i e e&voltare%

La om +et al T$ornton%' 4::-7 Manso"r et al%' 4::/' maim"=e +g et al%' ,--,7 C$en et

al%' ,--/ >i a #orcilor +Rat1y et al%' ,--5/ ov"l"l"i #ic19"# a fost efect"at 5 $ "#;

gonarotro#$in corionic; "man; +$CG/ #entr" inectare% Maoritate e ovocite colectate

la acest interval se afl; n fa&a e mat"ritate sa" metafa&; II >i s"nt evelo#mentally >i

meiotically com#etente% C" toate acestea' la ca#rine' ovocite colectate 5 $ "#;

inectarea $CG se afl; nc; n staii imat"re >i s"nt mat"rat in vitro #entr" ,D e ore

nainte e a a"nge com#eten=; meiotice +Jalassarre et al%' ,--57 Ra$man et al%' ,--Db'

,--./% (;strarea n veere acest l"cr" recenta noastr; c; ami D, $ FSB #ost' #l"s tratament c"

$CG ar #"tea fi #rotocol"l e #referat #entr" a o#timi&a #ro"ctivitatea e ovocite e

calitate #entr" IVM >i embrioni ICSI la ca#re% A>a c"m ea am men=ionat mai evreme's"rs; e abator ovar este foarte limitat; n Malae&ia' #rin "rmare' LO() ar #"tea fi o

#roce"r; alternativ; #entr" sa" e ca#r; s; efect"e&e IV(' mai ales ICSI st"i"%

Cronologia e a g;si semnificative e e0#erimente LO() la ca#rine este escris in

tabel"l %%%%


9/22

JASIC LO() TECBIP)E

A##ro#riate tec$ni*"e is a m"st for efficiency in LO() an finis$ing t$e #roce"re in a

minim"m time !it$ minim"m stress an iscomfort to t$e animal% (ro#er #ositioning an

restrain of t$e goat is very im#ortant for ac$ieving t$e o#tim"m access an e0amination

of t$e re#ro"ctive tract +Eey' 4:.5/% First of all' t$e goat s$o"l be anaest$eti&e !it$

#ro#er anest$etic agent an t$e anest$esia s$o"l be maintaine "ntil finis$ing t$e

s"rgery% T$is can be ac$ieve !it$ intram"sc"lar aministration of Qyla&ine

$yroc$lorie +Ili"m Qyla&ine9,-7 Troy Laboratories (ty Lt%' A"stralia/ at a ose rate of

-%,, mg 1g94 boy !eig$t follo!e by intram"sc"lar aministration of 6etamine

$yroc$lorie +6etamil7 Troy Laboratories (ty Lt%' A"stralia/ at a ose rate of 44 mg

1g94 b%!t% +Rosnina et al%' 4::,/% T$e anaest$eti&e goat s$o"l be restraine on a

stanar la#arosco#y table !it$ tilting a##arat"s at - egree angle !$ic$ !as first

evelo#e by B"let an Foote +4:./ !it$ $ea9o!n in a orsal rec"mbent to allo! t$e

g"t an Oment"m ma"s to move cranially%

LO() m"st be #erforme in a clean' "st9free area% Follo!ing t$e in"ction of

anest$esia an restraining of t$e goat' it s$o"l be s$ave ase#tically "sing a ra&or an

isinfecte by a##lying et$anol +:-/ over t$e s$ave #art of t$e abomen% Ins"fflations

of carbon io0ie' nitro"s o0ie an filtere air s$o"l be "tili&e from t$e a"tomatic gas

ins"fflating "nit an t$en connecte to t$e abomen of t$e goat to create a

#ne"mo#eritoni"m for vis"ali&ation of viscera incl"ing "ter"s an ovaries an to

maintain a constant intra9abominal #ress"re an air s#ace% T$e #ne"mo#eritoni"m


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s$o"l be create caref"lly to avoi over inflation !$ic$ can ca"se iscomfort of t$e goat

+C$eminea" an Cogni23' 4::4/% From e0#erience' it is seen t$at a##ro0imately to L

of carbon io0ie gas is re*"ire for an average a"lt goat +5- 1g b%!t%/%

T$e critical #art in la#arosco#y is t$e site of t$e trocar9can"la insertion% T$e trocar9can"la

insertion site s$o"l be a##ro0imately , cm cranial to t$e mammary glan an cm

a!ay from t$e miline in eac$ sie of t$e abomen in an average si&e goat% Once

#ro#erly inserte into t$e inflate abominal cavity' t$e trocar s$o"l be remove from

t$e can"la s$eat$ an re#lace by t$e telesco#e% Vis"ali&ation of t$e "rinary blaer

inicates t$at t$e o#erator is #roceeing !ell%

La#arosco#y is starte by t$e a##ointe s"rgeon an oocytes are retrieve by follicle

fl"s$ing an as#iration met$o% After recovery of re*"ire oocytes' t$e LO() is

terminate by ca"tio"sly removing la#arosco#e an t$e #air of force#s from t$e

res#ective can"la s$eat$% T$e ins"fflating gas is evac"ate from t$e abominal cavity by

man"ally #ressing t$e abominal !all t$ro"g$ t$e can"la $ole% After t$e la#arosco#ics"rgery' t$e incise s1in is s"t"re an antise#tics are s#rea over t$e s"t"re% LO()

#roce"re $as been s$o!n in

TEBICA KE JA LO()

te$nic; aecvat; este o necesitate #entr" eficien=a n LO() >i terminn c" #roce"ra

ntr9"n tim# minim e stres >i isconfort minim #entr" a animal"l"i% #o&i=ionare corect;

>i restr


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e0aminarea tract"l"i e re#ro"cere +Eey' 4:.5/% Mai nti =inerea e ca#r;' ar treb"i s; fie ras ase#tic' folosin "n a#arat e ras >i

e&infectate #rin a#licarea etanol +:-/ fa=; e ras #arte a abomen"l"i% Ins"fflations e

io0i e carbon' o0i e a&ot >i aer filtrat ar treb"i s; fie "tili&ate e ga&e a"tomate

ins"fflating "nitate si a#oi conectate la abomen e ca#r; #entr" a crea "n

#ne"mo#eritoni"m #entr" vi&"ali&area viscerele' incl"siv "ter"l"i >i ovarelor >i s;men=in; o #resi"ne constant; intra9abominale >i s#a=i"l"i aerian% (ne"mo#eritoni"m ar

treb"i s; fie creat c" aten=ie #entr" a evita as"#ra infla=iei' care #oate #rovoca isconfort

e ca#r; +C$eminea" >i Cogni2 3' 4::4/% Kin e0#erien=;' se vee c; a#ro0imativ 9 L

e ga& e io0i e carbon este necesar; #entr" o ca#r; a"lt mei" +5- 1g b%!t%/%

(arte critice n la#arosco#ie este site9"l e inserare trocar9can"le% Site9"l insertie trocar9can"le ar treb"i s; fie e a#ro0imativ , cm cranian; a glanei mamare >i cm istan=; e

linia meian; n fiecare #arte a abomen"l"i ntr9o ca#r; e imensi"ni meii% Oat;

intro"s; corect n cavitatea abominal; "mflate' trocar ar treb"i s; fie scos in teac;


12/22

can"le >i nloc"it e telesco#"l% Vi&"ali&area ve&icii "rinare inic; fa#t"l c; o#erator"l se

esf;>oar; bine%

La#arosco#ia este #ornita e catre c$ir"rg n"mi=i >i ovocitelor s"nt #rel"ate #rin nro>irea

fe=ei folic"lare >i metoa e as#iratie% K"#; rec"#erarea e ovocite necesare' LO() este

re&iliat #rin eliminarea #reca"=ie la#arosco# >i o #erec$e e force#s in teaca can"le

res#ective% Ke ga&e ins"fflating este evac"at in cavitatea abominal; #rin a#;sarea

man"al #eretele abominal #rin ga"ra can"le% K"#a o#eratie la#arosco#ica' #ielea

inci&ate se s"t"rea&; >i antise#tice s"nt r;s#


13/22

Fig% 4 Oocyte recovery from oestr"s sync$roni&e an

s"#erov"late goat ovaries t$ro"g$ LO() #roce"re% +a/ S"rgeons are con"cting LO()

#roce"re' +b/ follicle #"nct"re an oocyte as#iration an +c/ collection of follic"lar

contents incl"ing oocyte in t$e collection t"bes


14/22


15/22

KETERMIATIO OF P)ALITY AK GRAKIG OF RETRIEVEK OOCYTES

In any in vitro oocyte mani#"lation #rotocols incl"ing ICSI re*"ire t$e assessment of an

oocyteNs #otential to "nergo fertili&ation an f"rt$er evelo#ment #rior to s"becting it

to t$e #roce"re of interest% T$is generally involves eval"ation of t$e mor#$ology of t$e

c"m"l"s investment' t$e oo#lasm an t$e COC as a !$ole% Vis"al assessment of

mor#$ological feat"res remains t$e most im#ortant criterion for selection of COCs before

mat"ration to select evelo#mentally com#etent goat oocytes "ring oocyte retrieval%

T$erefore' oocyte classification or graing an selection become an im#ortant an

essential #art of any IV( #rogram% Li1e ot$er omestic or r"minant s#ecies' goat oocytes

are also classifie before being c"lt"re in t$e IVM mei"m% Bo!ever' classification or

graing of oocytes varies e#ening on t$e ty#e of e0#eriments an interest of t$e

researc$ers% $ile some researc$ers grae oocytes accoring to t$e C"m"l"s Cell +CC/


16/22

layers or com#actness of t$e CCs7 ot$ers grae t$em on t$e basis of mor#$ology of t$e

oocytes or oo#lasm' $omogeneity of t$e oo#lasm' iameter of t$e ovarian follicles from

!$ere oocytes !ere originate or iameter of t$e oo#lasm' or combination of CC layers

or com#actness of CCs #l"s mor#$ology of t$e oo#lasm% In cattle' goo res"lts !ereac$ieve from an oocyte $aving com#act' com#lete an m"ltilayere CCs' a finely

gran"late an $omogeno"s oo#lasm an an oocyte iameter meas"ring more t$an 4,-

?m +Ganolfi et al%' 4::D/% In a recent st"y in o"r laboratory' it !as fo"n t$at oocyte

*"ality im#rove !it$ mean #ercentages of goo *"ality oocytes +Grae A/ from ,:%D to

5D% !$en LO() !as #erforme at 5 an - $ #ost FSB #l"s $CG treatments7

elaying LO() to D, $ #ost FSB #l"s $CG i not im#rove t$e mean #ercentage of goo

*"ality oocytes% Oocyte *"ality an evelo#mental com#etence can also be relate tofollicle si&e an@or t$e com#osition of follic"lar fl"i or FF +Maison et al%' 4::,7 (avlo1

et al%' 4::,7 Cro&et et al%' 4::87 Ba&eleger et al%' 4::87 Arlotto et al%' 4::/% Bo!ever' t$e

etermination of t$ese #arameters is labore intensive an $arly com#atible !it$ t$e

ro"tine #ro"ction of a $ig$ n"mber of embryos% In fact' t$e assessment of follicle

iameter re*"ires t$eir issection from t$e ovary #rior to t$e isolation of COC an t$e

analysis of FF is generally base on raioimm"noassay #roce"res +Ganolfi et al%'

4::D/% Some im#ortant oocyte graing9relate st"ie in goat $ave been s"mmari&e inTable ,% Alt$o"g$' after retrieval oocytes are grae' $o!ever' t$ey are not al!ays

s"becte to IVM' IVF@ICSI or IVC accoring to graes% In t$eir st"y' Uim23ne&9

Maceo et al% +,--/ grae t$e #re#"bertal goat oocytes accoring to iameter after

IVM an st"ie t$e evelo#mental com#etence t$ro"g$ ICSI an IVC% T$e relations$i#

bet!een follicle an oocyte si&e +iameter/ #reicts t$e f"rt$er evelo#mental

com#etence of an oocyte% Oocytes retrieve from , mm si&e follicles are smaller in

si&e an less com#etent t$an t$ose of , to mm follicles +Cro&et et al%' 4::8' ,---/% In

most of t$e in vitro st"ies' alt$o"g$ c"m"l"s9free oocytes +CFOs/ are grae b"t

iscare an only COCs !it$ fe! layers of CCs are "se for IVM% Till to ate' t$ere is

no re#ort of IVM' ICSI an IVC in goat accoring to oocyte graes% Kifferent graes of

LO()9erive goat oocytes $ave been s$o!n in Fig% ,a9g%


17/22

Alt$o"g$' "ncommon in animals' !it$ fe! e0ce#tions' some researc$ers !or1ing !it$

$"man IVF or ICSI classify oocytes !it$ ysmor#$ic oo#lasm or ot$er forms of

ysmor#$ism to st"y t$eir evelo#mental com#etence an ca#acity to #ro"ce offs#ring

"sing ICSI tec$ni*"es% A s"rvey of available literat"re co"l not generate any ata

regaring classification or graing of ysmor#$ic oocytes% T$is may be "e to t$e fact

t$at as goo *"ality goat oocytes are reaily available from abattoir so"rce' t$erefore'

researc$ers are not m"c$ intereste to st"y t$e evelo#mental com#etence of

ysmor#$ic goat oocytes after IVF or ICSI% It co"l be note $ere t$at "nli1e ot$er

co"ntries' abattoir9erive goat oocytes are not reaily available in Malaysia "e to a

$"ge s$ortage of oes for sla"g$ter an t$e *"ality of abattoir9erive oocytes are also

lo!er "e to sla"g$ter of lo! *"ality oes% T$erefore' t$ere is a goo sco#e to st"y t$e

evelo#mental com#etence of ysmor#$ic goat oocytes erive from abattoir so"rce%

6ee#ing t$is in min' !e #ro"ce t$e first ICSI erive goat #reim#lantion embryos

from ysmor#$ic goat oocytes +Ra$man et al%' ,--Dc/% E0ce#t a recent one in goat' a

n"mber of re#orts escribing classification of ysmor#$ic oocytes $ave been #"blis$e in

$"man an a very fe! in bovine% Some of t$e im#ortant st"ies regaring classification

of oocytes $aving oo#lasmic ysmor#$ism mainly base on $"man an cattle oocytes are

#resente in Table 5% Kifferent ty#es of ysmo#r$ic goat oocytes $ave #ict"re in Fig%

5a9%

Table , Oocyte graing e0#eriments in goat


18/22


19/22

Fig% , Goat oocyte graing accoring to c"m"l"s cell

+CC/ investment an mor#$ology of t$e oocyte% +a/ Grae A' +b/ Grae J' +c/ Grae C'

+/ Grae K an +e9g/ Grae E% Grae A to K are selecte for IVM an Grae E are

iscare% Scale bar re#resents 8- ?m


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Table 5 Graing of ysmor#$ic oocytes in $"man' cattle

an goat


21/22

Fig% 5 Goat oocyte graing accoring to mor#$ology of

t$e oo#lasm% +a/ ormal oocyte' +b/ #ale oocyte' +c/ oocyte !it$ big fat glob"les an +/

ar1 oocyte% (L W (ale oo#lasm' FG W Fat glob"le' K W Kar1er &one% Scale bar

re#resents 8- ?m

COCL)SIOS

Li1e cattle' goat IV( incl"ing IVF an ICSI !ill become first emerging ART in t$e

livestoc1 in"stry in near f"t"re% T$e #resent state of goat IV( s$o!s a great eal of

#romise% T$ro"g$ im#rove n"trition an veterinary assistance' ramatic c$anges $ave

been bro"g$t in t$e management of goat breeing "ring t$e last t!o ecaes% Bo!ever'

es#ite t$ese im#rovements' AI is t$e only ART !iely a##lie in selection #rograms till


22/22

no!% T$e ot$er im#ortant tec$ni*"es relate to goat IV( for e0am#le s"#erov"lation an

estr"s sync$roni&ation' !$ic$ are essential re*"irements for LO()' still $ave a large

margin for im#rovement% As bot$ *"antity an *"ality of recovere oocytes are cr"cial

for IV( o"tcome in goats' t$erefore' more researc$ s$o"l be foc"se on LO()tec$ni*"e for OR from live goats !$ic$ !ill in t"rn im#rove t$e efficiency of t$ese in

vitro tec$nologies% Alt$o"g$ abattoir9erive ovaries are a c$ea#est so"rce of oocyte'

$o!ever' in Malaysian sit"ation LO() is t$e most reliable an efficient met$o of OR%

Jeca"se LO() is #erforme in t$e live onor' t$erefore' OR rate an *"ality of oocytes

are m"c$ $ig$er t$an abattoir ovaries as same goat can be "se for several times%

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