HOI NGHI KHOA HQC C O N G NGH? SINH HOC T f . ,?,
NGHIEN CLTU HE T H O N G CONG NGHE P H C I VA NHAN B A N V 6 TINH PHUC
VU NHU CAU CAl TI^N NANG S U A T D O N G VAT NUOl VA B A O T O N C A C L O A IDONG VAT HOANG DA
Bill Xuin Nguyen^*, Nguyin Thi l / d v \ Duvng Dinh Long^, Nguyen Khic TEch', Do Tiroc^, Phan Ngpc Minh^ BitI Linh Chi\ Nguyen Trung Th^nh^, Nguyen Vift Llnh^, Nguyin Van Hanh'', Dgng Nguyin Quang Thdnh , Nguyin Thj M l n \ Tr^n Tht Them', Nguyin Thj Nhung', Nguyen Thj Hi$p', Nguyin Thi Hong', Nguyin Vdn LSm . Wiin Cdng nghi sinh hgc. Vign Hin lim Khoa hgc vi Cong nghe Viet Nam
^Trung tim COng nghg sinh hoc Dong Nam A
^Vign quy hogch vi phit triin, Bg Ndng nghiip vi Phit triin ndng thin
T 6 M TAT
iJng dyng c6ng nrfi? sinh san Men dai, mmg dd c6 c8ng ngh? phoi va nhan ban v6 tinh (NBVT) la giai phap ky diu|t d^ic bigt c6 h i ^ qua de cai tirai nSng suit vat nu6i, dfing thcri la mpt cong cy ti6m nang trong vifc bao ton vi tai t?o ngu^n gen cSc lo&i d^ng v|I bj de dQa tuy^ chiing, Trong bao cdo nay, chung tdi trinh bay y tudng phit trign HB th6ng nhan gi&ng H^t nhSn Md-Cly phdi di/a tr€n Icit quii nghien clhi c ^ cong ngh^ gSy dOng due d^ng loat, gay rung trtmg nhieu, thy tinh dng nghidm, dong Ipnh phdi, x^c djnh gioi tinh phdi bing vi phlu diuat vi PCR, ciy phoi vi chSn doan co that s<^ da dugc thvtc hi$n thinh cdng tren cic giong bo Laisind, Holstein, Gir vi bd Ha-AtL Bio cio cung dl c|p d^n kk qui nghiSn ciiu quy trinh NBVT bing c§y nhan cimg loai vi Idiac loii da duoc ti^n hinb nhim mgc dich xay dung co sd cdng nghf cho vice bio thn dc loii dOng v$t qu;^ hi^m nhu bd Kouprey, Sao la, lgn mini ban dja. Cic md hlofa ciy nhin khic nhau da dugc ngbien cim, bao gdm ciy nhan tk bio sioh dudng cila bd Gh vio tnmg b6 Laisind da loai nhan, ciy nhan t£ bio sinh dudng cua Saola vao trung bd Laisind dS lo^ nhin vi cay nhan te bio sinh dudng ciia Iqm Bin vio trimg lgn Landraee da lo^ nhan. Ti If phoi nhan bin phit triin tdi giai d o ^ phdi nang tuong duong cho cac md hinh tren li 23,7%, 27,7% vi 12,5%. Tom lai, cic cdng nghf sinh san va nhan bin vd tinh da dugc nghi&i ciiu vi c h u ^ bj dl dip iing nhu ciu phit tnIn Hf thong nhin gidng H^t nhan Md-Ciy phoi doi vdi din bd thjt-siia va bio ton kjp thdi nguon gen da d^ng suih hgc df ng v§t
TuritAda: bd sira, cdng nghe phdi, h^ nhan giong hat nhan md, lgn mmi, Kouprey, nhan bin vd tinh, Saola.
M^DAU
Cong ngh# phdi vi t l bao goc dang trfr thdnh hu'crng nghien cO'U cd inh hu-dng s3u r$ng do! vd'i sy phat triin y hgc trong thi ky 21. Gidi Nobel sinh ly vi y hpc cic nim 2010, 2012 da du'g'c trao t^ng cho cac nhi khoa hpc di tien phong tnsng Finh vyc dilu trj vd sinh bing thy tinh 6ng nghidm, nghi§n cO'U nhin bin vd tinh (NBVT) v i tgo t l bio g6c da nang.
Anh hirdng cda cic cdng ngh^ nay cung dang du^c phit huy hong cic ITnh vyc chan nuoi d$ng v§t v i bio tdn da d^ng sinh hgc. H$ th6ng chgn lgc nhin giong theo nguyen tac ket hgp cac hogt d5ng kilm tra tien d9 di truyen v i cdng nghO phdi hi$n d^i da duvc hiln khai it hiu het cic hung tdm giong gia sOc h i n thi giiri (Nicholas, 1996). Ngay sau sy ra ddi cua Dolly, d§ng v^t nhin bin diu tiin, cong ngh? NBVT d i du'pc il-ng dyng thinh c6ng d l bao ton cac giong bd v i dfl dang tren bfir tuy§t chung (Wells et al., 1999). Thanh cdng cua Waykayama et al. (2006) trong vi^c tao chuOt NBVT tir ca till duvc luu gio 16 nam trong tii Ignh cho Uily vigc bio quin ngu6n gen d l t i i tgo loii d i cd Uil thyc hi^n tgi tit ci cac cc sd' bao ton da dgng sinh hgc tren thi gi6i.
Trong bii niy, chOng tdi se binh biy tSng hpp cic kit qua nghien curu cdng ngh? phoi va NBVT duvc thyc hi^n tgi Vi$n Cdng ngh^ sinh hgc, Vign Han lam Khoa hgc v i Cdng ngh# Vidt Nam trong giai dogn tCr nim 1960 t6i nay v i khi nSng t6 chi>c li-ng dyng chiing vd'i hai myc dich: 1) Xiy dyng H§ thing nhan gilng Hgt nhan Md'-Cay phdi nhim dip tjng nhu ciu phat trien nginh dian nudi bd thjt-sQa Uieo huVng higu qua v i bin vi>ng; 2) Xiy dyng cac quy trinh cdng ngh$ Sinti sin v i NBVT c i n thilt cho vi$c bio ton kjp thd'i cic loii ddng vit dac hO-u cd nguy co' tuy^t chCing cao ir Vi$t Nam nhir bd Kouprey, Saola, lyn mini bin dja.
CdNG NGHg PHOI V A H ^ N H A N GidNG HAT N H A N tAC/
Hg Nhan gidng Hat nhin Md - Giy n,ing ht>ng nhilu v i cly phfii (ONBS-MOET) la ket qua ciia sy kit hpp cic ho^t ddng chpn Igc nhin giong vol cdng nghg phdi nham tao ra hidu qua cii tiin tiin dd di truyin cao nhit dli vd'i ddng v^l nudi.
V l phu-ong dign ky ttiuat, Hg Nhin giong Hgt nhin Uit bao gim cic khau: 1) Chpn Igc ca t h i dO tiiu chuin cho v i nhfln phdi; 2) Sin xuit phdi in vivo bang cic ky Uiugt giy njng tning nhilu v i thy tinh nhin tgo; 3) Sin xuit phdi in vitro bing cic ky thugt nudi thinh thyc Uijng v i thu$t thy tinh trong Ing nghidm; kit hpp thu tn>ng bang ngi soi v i Uiy tinh Ing nghidm; 4) Sin xuat phdi xac djnh giai tinh bing ky thuyit vi phlu thudt kit hyp PCR hoac thu tinh 6ng nghi0m v&i tinh tiling d i phin logi gidfi tinh; 5) Bao quin, ddng lanh tinh, phdi cCia cic c i Uil thudc nhom con gilng cao d p ; 6) Giy ddng dye ding logt v i ding pha giija ca thi cho v i nhgn phdi; 7) Cly phfii, dian doin cd chi>a sdm, theo doi sinh dS;
8) Dinh gii tien dp di tnjyIn, chpn Ipc c i the ddi sau.
V l phu'ang di|n quin ly va to chi>c tnln khai, Hg Nhin gilng Hgt nhin Md bao ^om cic hogt dfing thu$c khu vyc trung tam v i hogt ddng thu$c khu vyc ngogi bien. Hogt ddng trung tam cd nhigm vy bien khai cic khiu ky thuit sinh sin h i ^ dgi v i dinh gia tiin dfi di truyin nhim tgo nguon hgt nhan cic con gilng hgng cao cap va ngan hing tinh. phfii^ola ' chimg. Hogt dgng ngoai bidn cd nhiem vy phat tin v i luin chuyin hai chilu cic kit qui chgn Ipc di tnjyen v i ca thi dW sau gi&a khu vyc tmng tam v i khu vyc ngogi bien thfing qua cic ky thugt dan giin cd thi hiln khai d di$n rOng nhir thy tinh nhin tao, cly phfii dfing Ignh, cung cip con dye giong.
HOC C O N G N G H f SINH HOC TOAN QUOC 2013
Nghien ci>u iing dyng c i c cfing nghe sinh san tren c i c giong bd thit-sOa nudi d Vi|t Nam da duvc tiin hanh trong giai dogn h> 1930 tdi nay nhim chuin bj cdc quy hinh cong nghg d n ttiilt cho vide xay dyng He Nhin giong Hgt nhan Md, t>aog6m:
1- Quy frinh sir dyng thich hgp cac chit (Slu khiin sinh t) sinh sin (PMSG, LH, SimchroMate B, GnRH, Eshogen, PG) a l g i y ddng dye ding pha. g i y rung tning nhieu an toan v i higu qua d l i vin cdc giong bd ving, bd Laisind, bo Ha- An v i bd sOa Holstein.
2. Quy hinh si> dyng cdc ky thudt nudi thdnh thuc tiling, thy tinh Ing nghidm va nudi phdi in vitro d l s i n xuat phfii cd chit hfyng di fruyin duyc cai tiin.
3. Quy frinh vi phau thuit hiit t l tido ph^ v i si> dyfng cdc cip moi dgc higu cua nhilm sic t h i Y (INRA1, INRA2 ) d l xie dinh gidi tinh phdi bing ky ttiuat PCR (Bui et al., 1999).
4. Quy tiinh bao quin vd ddng Ignh phoi tiao gim cdc phirang phap bdo quan Ididng ddng lanh nham phyc vu nhu cau vdn chuyin vd di chuyin phdi trong thdi gian ngin, phirang ptiip bio quin d i i hgn phfii bing dfing ignh nhanh hogc ddng Ignh cyc nhanh (Nguyen etal.. 2000;2004).
5. Quy trinh td chiic d y phfii tgi khu vyc ngogi bidn bao gfim c i c hoat ddng d y ptidi d quy md chin nudi gia dinh nhd vi phin t i n tgi lang x i , quy md hyp t i c xa chin nuoi tir nhan vd quy md ti^ng trgi quoc doanh.
Cdc thfing so ccr b i n lien quan k i t qua nghien cdu c i c quy binh tren diryc trinh bay d Bang 1.
B^tg 1. Chi tieu vi ket qua sin xuit v i cly phdi bd trong dilu ki^n chin nuoi chira tieu chuin hda.
Chl tidu theo ddi K4t qui cd ttie dat Phin ling ryng tning nhilu 8,0 - 9,6 tning ryng/ con/ iln
Tl 1$ phdi cd thi d y 70%
Tl 1$ tning thinh thyc in viho 60% dli vdi tning bd laisind, 85 - 87% ddi vdi tning bd Holstein TT Id hiing thu tinh v i phan chta 68% ddi vdi hiing bd laisind, 78% ddi vdi tning bd Hotstem Tt 1$ phdi TTON phit hien den giai dogn phdi diu-phdi 52% ddi vdi phdi bd laisind. 58% ddi vdi phdi bd Holstein nang
Ti 1$ phdt TTON ptiit tnln d ^ giai do^n phdi nang 19% ddi vdi phdi bd laisind, 32 - 38% ddi vdi phdi bd Holstein Tl 1$ bd ddng dye ding pha frong vdng 6-10 g 72% d& vdi bd ving v i bd Laisind, trdn 87% ddi vdi bd Hi-ln v i bd
Holstein TI Id phdi an toin sau vi phlu ttiuit hdt t l bao > 95%
Miic dd phin logi gidi tlr^ phdi ttieo kit qui PCR chinh xie trdn 95%:
Thdi gian bio quin ngin hgn phdi Tii da 4 ngiy
Ti Id ptidi sing sau ddng Ignh trong ni ta Idng v i giai ti^n 90% di» vdi phfii NBVT, trdn 80% ddi vdi phfii TTON ddng
Ti 1$ thg thai vio ttiing ttiii 3 sau d y phfi 60% ddi vdi bd laisind, 60 - 75% d6i vdi bo H i ^ v i Holstein.
Ti t$ smh t>§ con sau d y phdi 45 - 50% ddi vdi bd Laisind. > 55% ddi vdi bd Hi-An
Cic ttil nghidm d y phdi diln hinh da diryc thyc hign de tgo bo dye gilng Charolaise bang d y phdi ddng ignh v i o bo nhdn ttiupc giong Ha -An (Hinhl.b), bd dye gilng Gir bing d y phdi TTON khdng dfing Ignh v i vin chuyin xuydn lyc ffja vio bd nhdn ttiufic gilng Uisind (Hinh l.c). bd sira Hdstein bang d y phdi TTON v i o bd nhin gilng Laisind (Hinh l.d), v i bd sinh ddi do cdy phCa TTON v i chpn gidi tinh vao bo nhgn Laisind (Hinh 1 .e).
Kit qui nghien ciiu v i cdc ttidng s6 ky ttiudt ttiu diryc cho ttiay v i ^ Cmg dyng cic quy frinh cong ngh^ TTON. xac ^nh gidi tinh v i d y phfii d l sdn xuat con gilng d i l t lugng tot la kha thi v i cd thi friln khai hong dilu kign chin nudi hi§n nay.
C 6 N G NGHE PH6I V A N H A N B A N V 6 TiNH CAC LOAl DONG VAT HOANG DA VA QUt HI^M
Cong nghd NBVT duvc phat frien frdn ca sd kit hyp cde ky ttiudt sinh san ca ban (nudi ttidnh ttiyc tning, TTON, nudi phdi v i d y phfii) vdi ky ttiuat tgo phfii nhin bin vfi tinh.
Bin chit cue NBVT Id si> dyng cdc yeu to cd kha ndng tdc dgng qua ca ehl di fruyin ngogi vi d l ma hda lai chuong trinh hogt dfing eOa nhin te bao sinh duong ttieo hudng phat hiln cua t l bio phdi. Trong trirfing hap ting dyng cdng nghd NBVT vio myc dIeh bao ton v i t i i tgo edc loai ddng v^t bj de dpa hjygt chiing, qua hinh md hda Igi dirp'C thyc hidn ttdn ea sd si> dyng ky thuit d y nhan khdc loai
Bd xdm Kouprey [Bos sauveli) l i gilng bd rtrng duy nhit chira duvc nghidn cdu diy dd va dang bj de dpa hjygt chiing (Tbeo T l chlic bao t i n qudc t l lUCN. mpt nhdm nhd khoang 50 ca ttil cd ttie cdn ton tgi frong eac khu vyc giira Nam Vi#t Nam. Lio vd Campuchia).
Saola {Pseudoryx Nghetinhensis). Id loii ddng vgt ed vtj mdi duve phdt hi§n d Vidt Nam v i Lio v i dang bj xep hg^ng bj de dpa hjy#t chung mdc dd cao nhit. Theo ket qud khio s i t 2010, so luvng Saola giam nhanh tir han 3000 ca t h i vio liic mdi phdt hidn xudng cdn khoing 200 ed ttil (Nguyen et al., 2007).
Kit qua nghidn ciiu phdn logi dya trSn eic chi tidu hinh ttiii vd sinh hpc phan td (Thanh e( al.. 2005) cho Uily Kouprey v i Saola diu ttiufic hp Bovinae, dilu do cho phip ttiam dd kha ndng sd dyng tiling cua cie loii frdu, bd va de ciru d l san xuit phfii Saola v i bd Kouprey thdng qua ky thuit d y nhdn khic loai.
HOI NGHI KHOA HOC CONG NGH6 SINH HOC TOA
r ]
-mw^-
Hinh 1. a- Vi phlu thugt hiit te bao phdi TTON. b- Ket qua ciy phoi Charolaise ddng lanh vdo bd nhan Hi-An. c- Bd Gir sinh ra do cly ph« TTON, inh luc 24 thing tudi. d-Kdt qui cly phdi bd Holslein vao bd Laisind. e- Bd sinh ddi sau cay phdi xie i^nh gidi tinh. f- Phol bd NBVT. G- Phdi Saola NBVT. i- h- Hinh kinh hien vi quet cua phdi Saola sau xCr ly aitti-BrdU antibodies.
Cdc md hinh tgo phdi khac nhau d i duve tiin hanh vdi quy trinh NBVT bao gom cdc birdc: 1) Thu miu, nhdn nufii vd Idp ngdn hdng t l bdo cua dfing vit d n nhan ban; 2) Thu buong tning ti> Id m l , nudi ttidnh ttiyc, chgn Igc kilu tiling thich hyp cho cac thao t i e logi nhdn vd dung hap ; 3) Giai dfing vd chuin bj t l bio d l d y nhdn; 4) Thao tic d y nhin, dung hyp nhan t l bdo vd hiing: 5) Chpn lyc to hap tning-tl bdo d i dung hop, kfch hyp giai dpgn hogt dfing cCia nhin;
6) Chpn Ipc phdi. nudi phdi NBVT.
Thi nghigm chgn lgc kilu hiing da diryc thyc hidn vdi cic mfi hinh d y nhan t l bio friu vio tning bd v i nguvc Igi (tl bio bd-frdng trau). Kit qui thi nghigm eho thiy friing bd co uu t h i han so vdi triing ti^u ve phirang didn ^ a o tdc cay nhdn vd kha nang dung hcrp (ti Id triing nguydn vgn sau loai nhan v i fi l§ friing dtpyc dung hyp l i 74% vd 60% dli vdi friing bd, 51% vd 32% doi vfin frdng trdu).
Md hinh d y nhin te bio sinh dudng cua tid Gir vio t l bio tning tid Laisind da logi nhin dd duyc su dung vdi myc dfdi chuan bj quy frinh cho vide tgo phdi bd Kouprey nhin ban frong tuang lai. Trong frtrdng hyp Saola. t l bao sinh dudng, bao gim e i t l bdo &iu duyc vdo liic Saola dang sing sau khi bj mic bay cda ttiy sdn v i ttiu v i o thdi diem 10 gid sau khi Saola chit, d i duoc sd dyng d l d y nhan
Kit qua thi nghigm cho thiy cd the su dyng tning bd Laisind d l tao phdi bd Gir (Hinh l.f) vd phfii Saola (Hinh Ig) nhin bin cd tiin dfi phit friln binh thufing, dgt fi Id 25-27% phfii nang vio ngdy tuoi ttid 7 (Bing 2). Kit q u i nghiin cdu phin ung ciia phdi vdi anfi-BrdU antibodies bang kinh vi qudt (Hinh 1g. Ih) cho ttily khfing c6 sy khac bigt v l hogt ddng phidn m i gen gida hai nhdm phdi Saola/bfi v i Bd/bd (Bui etal., 2002).
n v i i N u n i M M U A M O C C O N G N G H f S I N H H O C T O A N Q U O C 2 0 1 3
B i n g 2. Anh T l b i o cho
Bd Saola Lyn B i n
htfdmg cda kieu t d h ^ p t l b i o CHO-tn>ng len k i t qua d y nhan Tning loai n h i n
Bd Bd L y n l a t C N
V Id dung hyp (%) 60,67 53,21 72.70
TTI$ p h i n chia (%) 79.12 78.92 81.20
Ti Id phdi d i u (%) 27.5 39,15 50,00
TI le phdi nang (%) 23.07 27.71 12.50 T h d n g h i d m d y phfii b d N B V T vdo b d n h d n Laisind d q u y m f i n h d rfio k i t q u i 4 2 % bo c d c h u a v i o ttiang I h u 3 sau cdy phdi, 5 % b d d u y h i h a n g ttidi p h i t friln binh ttiudng cua ttiai d i n ttiing ttid 7.
Trong t r u d n g h y p l y n mini bdn dja, n g h i i n c d u N B V T vcri m d hinh d y nhdn t l b i o sinh d u d n g cOa l y n B d n vao tiling tidng l y n Landraee da loai n h i n cho k i t q u i h e n 7 0 % frdng d u y c d u n g h y p sau d y n h i n , 5 0 % frdng p h i n chia d i n giai d o g n 4 - 8 te b i o , 1 2 , 5 % phfii phdt t r i i n d i n giai d o g n phfii nang (ttiap h a n so v d i ti Ig 1 6 % phdi nang trong t n r d n g hyp tgo phfii bang T T O N d l y n B i n v d 2 7 % Ipn Landraee) ( l / d c etal., 2008).
K£T LUAN
C i c q u y trinh cfing nghg s i n h s d n d n ttiilt cho vide frien khai H § n h d n g i l n g Hgt nhdn M d - C l y phdi d e d i tgo n i n g s u i t d i n b d ttijt-sua^da d u y c nghidn c d u d i n g bO, h i k h i u san x u i t phfii b i n g cdc k y t h u i t in vitro tgi khu v y c tiung t i m d i n k h i u t l c h l i c d y p h d i d k h u v y c ngogi bien, d i n g ttidi bao h d m cdc hogt dfing d c d c guy mfi q u i c gia va q u o c t l . K i t qud nghiSn c u u c h o p h d p k i t l u | n , v l p h u a n g dign ky ttiudt w d c t l c h u c Hg nhdn g i i n g Hgt nhdn M d - C l y phfii frong giai dogn hi$n nay l i phCi h y p vd k h i t h i .
Quy trinh N B V T va c i c cfing nghd sinh s i n lidn q u a n da d u y c nghidn c d u frdn cdc mfi h i n h c l y n h i n ttiieh h y p v d san s i n g vi§c t r i i n khai d n g d y n g d l b i o t i n c d c lodi dpng vdt c6 nguy c a tuydt chung cao d Vi#t nam n h u Kouprey, Saola, lyn mini.
TAI LI$U THAM KHAO
Bill Linh Chi. L i van Ty. NguySn ttij l/dc, Nguyin Huu Oiic, Bill X u i n Nguydn, LS Oinh Luwng (2000). Xdc djnh gidi tinh phdi d bd b4ng k9 thuit PCR. Tgp chl CX truyin hgc vd Ong diing 1:6-11.
Bui LC, Vignon X, Campion E. Laloy E, Lavergne Y. Ty LV, Nguyen BX, RenanJ JP (2002). Use of interepecies nucJear fransfer to shidy the early embryonic developnent and nuclear activities of the endangered species Pseudoryx nghehnhensis (saola). Thenogenology 57,427.
Nicholas FW (1996). Genetic improvement through reproductive technology. Anim Reprod Sci 42 205-214.
Nguyen BX, Sotomaru Y, Tani T, Kato Y. Tsunoda Y (2000). Efficient cryopresenration of bovine blastocysts denved from nuclear transfer with somatic cells using partial dehydration and vitnfication. Theriogendogy 53:1439-1448
Nguyen BX, Uoc NT, Ty NV, Monson RL, Leibfned-Rutledge ML, Due NH. Bui LC, Huu OX, Hanh NV. Thanh NT, Linh NV. RuUedge JJ (2004). PRXluctlon of fropical diary calves by embryo transfer using local Laisind (60s indicus) recipients and intercontinental fresh IVF shipped embryos. Reprod Pert Dev, 16 (1,2): 249-250.
Nguyen Bui Xuan. Do Tuoc. Nguyen Thi Uoc, Trung Thanh Nguyen. Bui Linh Chi, Hoang Ngoc Khanh, Nguyen Van Keo, Comizzoll P, Renanj JP (2007). Saola rescue plan with contributk>n of cryobanking and ART. Genetics and Applications, special issue on Biodiva- VAST Intemational Conference on « Reproduchve biotechnologies for consenralion of bkxliverslty and sustainable develiqiment », Hanoi, Dec. 17-18,22-25
Nguyen TT, Menefrey F, Genini S, Nguyen VL. Vogeli P, Nguyen BX, Stranzinger G (2005). Application of bovine microsatellite markers for genetic studies on Saola (Pseudoryx nghetinhensis) compared to six different species of the family Bovidae. J Anim Breed Genet 122(3):195-8.
Uoc NT, Ty LV, Due NH, Long DD. Nguyen BX (1995) Gdy dgng dye ddng pha va chuan bj con nh§n phdi d bd LaiSind Ky yeu Vien Cdng ngh$ Sinh hgc. NXB. Khoa hgc v i Ky thu$t. 179-185
Uoc Nguyen Thi, Le Van Ty, Nguyen Huu Due, Bui Linh Chi, Nguyen Tmng Thanh, Nguyen Viet Linh, Nguyen Van Hanh, Quan Xuan Huu, Nguyen Thuy Anh, Hoang Nghia Son, Duong Dinh Long, Bui Xuan Nguyen (2003). San xuat be sOa bang thy hnh dng nghiem v i c l y phfii xdc dinh gidi tinh. T i / li$u Hdi nghj Cdng ngh$ Sinh hgc todn qudc. NXB Khoa hoc va Ky thu$t, 717-719.
Nguyin Thi LTdc, Nguyen Vidt Linh, Nguyin V i n Hgnh, Q u i n X u i n H&u, D i n g Nguyen Quang Thinh, Nguyin Thj M i n . T r i n Thj Thom, Nguyin Tmng Thdnti. Biii Unh Chi, Duong Dinh Long, NguySn K h i c Tich. Phan Ngpc Minh.vi Bui X u i n Nguydn (2008). Sdn xuit phdi Ion mini n ^ dia b i n g t l hyp cdng nghd ™i9 nghifm va n h i n b i n vd tinh, CNSH-so chuydn san, 6 (4A), 625-635.
Wakayama S, Ohta H, Hikichl T, Mizutani E, Iwaki T, Kanagawa O, Wakayama T (2008). Production of healthy cloned mice from txidies frozen at -20 degrees C for 16 years..Proc Neil Acad Sci USA 105(45): 17318-22
Wells RJ, Misica PM, Tenrit HR (1999) Production ofcloned calves following nuclear transfer with cultured adult mural granulosa cells.
S/df/7eprDd 60.996-1005.
HOI NGHI KHOA HOC C 6 N G N G H E SINH HQC TOAK
DEVELOPING EMBRYO BIOTECHNOLOGY AND SOMATIC CLONING FOR IMPROVING ANIMAL PRODUCTIVITY AND PROTECTION OF ENDANGERED WILD SPECIES
Bui Xuan Nguyen''*, Nguyen Thj Uoc\ Dipong Dinh Long', Nguyen Khac Tich', Do Tuoc', Phan Ngoc Minh', Bui Linh Chl\ Nguyen Trung Thanh\ Nguyen Viet Linh\ Nguyen Van Hanh\ Dang Nguyen Quang Thanh', Nguyen Thi Men^, Tran Thi Thom\ Nguyen Thi Nhung\ Nguyen Thi Hiep\ Nguyen Thi Hong', Nguyen Van Lam^
'institute of Biotechnology, Vietnam Academy of Science and Technology
^Soueast Asia Biotechnology Center
^Forest Inventory and Planning Institute, MARD
SUMMARY
Application of reproductive biotechnology, including embryo technology and somatic cell nuclear transfer has been considered in^rtant approa^ to iiiq>rove animal productivity and protect endangered species from extinchwi. We present in this paper the results of invcstigaiion on the oestrus synchronization, in vivo embryo production by supnovulation and artificial insemJnatioD, embryo production by in vitro maturation and fertilization, embryo sexing by biopsy and PCR, emt>Tyo transfer and early pregnancy diagnosis which were earned out for dairy and beef cattle under tropical conditions of Vietnam. The possibility to establish a System of Open Nuclear Breeding using embryo technology was discussed basing on the suceessfiil production of Charolaise, Gir and Holstemcalvesbyin vitro fertilization and embryo transfer into local recipients of Laisind and Ha-An breeds. We also present herein the main results of research on developing technique of somatic cell nuclear transfer to meet the need of conservation and protection of inportant eadangered manunahan species of Vietnam and region such as bovine Kouprey (Bos sauveli), Saola (Pseudoryx nghetinhensis) and native minipigs. Different models of nuclear transfer including Saola somatic cells into Laisind enucleated occyte, Gir cattle somatic cells into Laisind enucleated occyte and Ban imnipig somatic cells into Landraee enucleated occyte were investigated. The rates ofcloned emt>ryo developed to blastocyst stage were 23.7%, 27.7% and 12.5% for the case of Gir, Saola and Ban minipig, respectively. In conclusion, the basic technique platform had been estabhshed for the application of advanced reproductive biotechnology for improving dairy and beef animal production and biodiversity conservation.
K^ words: Dairy catUe, embryo, Kouprey, minipig, ONBS-MOET, Saola, somatic cloning.
'Author for crrespondance: [email protected]
