NGUYEN VAN LY - Anh huong gdy rung trung ...
ANH HUONG GAY RUNG T R U M G 35 NGAV/LAN DEN SO PHOI /BO/NAM TREN DAN BO SLTA
Nguyen Van Ly', Luu Cung Khdnli\ Nguyen Thi Tlioa'\ Dd Vdn Hucnig', Tran Swn Hd\ Nguyen Thi Huirng'. Liru Nggc Anli^ vd Phan Le Sinr'
'Vu Khoa hoc eong nghe - Bo Nong nghiep & PTNT Phong Thi nghiem trgng diem Cong nghe te bao dong vat
Bo mon sinh san va thu tinh nhan tao Vien Chan nuoi - Thuy Phuong - Tii- Liem - Ha Noi
* Tac gia lien he: Nguyen Thi Thoa - Phong TN Trong diem te bao dong vat
Tel: (04)22.166.165/0977. 249.755; Fax: (04) 38.389.775; Email: methoa2002(gyahoo.com
ABSTRACT
Effect of superovulation interval 35 days/time on embryo number/eow/year of dairy cow Bo\ine embryo transfer has been studied and applied widely around the world. Superovulation and embryo recovery were the first important basic steps of embryo transfer technology. The objective of our study was to compare 35 day-reapeated to 2-3 months repeated superovulation and embryo recovery. Two high genetic \alue dairy cows were used as donors in this experiment. Cows received a standard 0\sync protocol plus a Prid for 12 days (starting on Day 0). From Day 10 to Day 13, cows were administered i.m, twice daily. Prid was removed and then donors received PGF2o on Day 12. On Day 14 and 15. donors showed oestrous were inseminated articificially twice. The number of copus lutea was counted and embryos were collected on Day 21, Donors received PGF2„ on the day of embryo collection. On Day 35 cows were given Prid, and subsequent steps were carried out repeately. In 8 experimental sessions, two cows reponsed well to the administration of hormone. We collected a total of 109 embryos and ova (including 88 transferable embryos) from 2 donors. The average number of embryo and ova/cow/ collection was 6.8. The average number of transferable embryo/cow/time was 5.5. Our results indicated that repeatability of 35 days- superovulation and embryo reco\ery increased the number of embryo collected per donor a year. Although the results of superovultion depend on many factors such as response of donors to the kinds of hormone using, ability of Al, flushing...,we can apply the 35-day repeated superovulation schedule to increase the embryo number per valuable donor per year in Vietnam conditions.
Key words: superovulation, repeated, bovine, embryo transfer, FSH, PGF2„
DAT VAN DE
Trudc day sau moi lin gay rung trirng nhieu (GRTN) phai de bd cho phdi nghi 2 - 3 thang rdi mdi tilp tue GRTN lap lai. Nhu vay, mot nam chi tien hanh thu phdi 3 - 4 lan va sd phdi binh quan/bd/nam chi dat 15 phdi. Hien nay cdng nghe cay truyen phdi va ky thuat gay rung trimg nhilu lap lai da cd nhilu cai tiln. Rut ngin thdi gian GRTN lap lai tir 2-3 thang xudng 35 ngay/ian/bd cd thi tang sd phdi thu dugc /bd/nam gdp phan nang cao hieu qua trong cdng nghe cay truyen phdi (CTP) bd.
Vi vay, chiing tdi tiln hanh dl tai ''Nghien cuu irng dung nit ngdn gdy rung triing nhiiu vd thu phdi lap lai tir 2-3 thdng xuong 35 ngay tren bo sira ".
VAT LIEU VA PHU^ONG PHAP NGHIEN CLfU Doi tu'ong nghien ciiu • '
Bd cai lai F2 hudrng sira, nang suat 4000-6000 lit/chu ky
Cac hormone sinh san: FSH, PGF2u, Progesterone (PRID, CIDR)
VIEN C H A N N U O I - Tap chi Khoa hgc Cdng nghe Chdn niid, - So 15 -Thang 12- 2008
Noi dung nghien ciiu i - Nghien cii'u gay rung trirng nhieu theo quy trinh 35 ngay/lan.
Danh gia phan irng ciia budng trimg bd qua cac lan gay rung trimg nhieu.
Danh gia ket qua thu hoach phdi va chat lugng phdi thu dugc.
Phuong phap nghien ciru Chgn bo cho phoi
Bd cho phdi la nhung con bd cai lai hudng sira khde manh. dugc chgn tir dan hat nhan.
nang suat sira binh quan 4000-60001/chu ky. cd chu ky sinh due binh thudng. sinh de binh thudng, khdng cd di lat o co quan sinh due, budng trirng mem, khdng mac cac benh truyen nhiem, cd do tudi tir 2-6 nam. Tien hanh kiem tra boat ddng ciia budng trirng bang phuong phap khani qua true trang.
Tieu chuan bo due phoi tinh
Bd GRTN khi ddng due dugc thu tinh nhan tao bang tinh ddng lanh. Tinh dich phdi gidng phai lay tir nhirng due gidng tdt nhat, dac trung cho gidng ve tinh trang mong mudn nhu: san lugng sii'a, san lugng thit, tinh trang cao san d due gidng phai dugc di truyen cho the he sau.
Phuang phap GRTN
Chiing tdi tien hanh GRTN lap lai tren bd sir dung hormone FSH va PGF26.
Bd sau khi dugc kiem tra qua true trang thay cd the vang ciia chu ky trudc, dugc dat PRID vao xoang am dao. Ngay dat la ngay 0 cua quy trinh
Tiem FSH tir ngay thii' 10 ke tir khi dat PRID trong vdng 4 ngay theo lieu giam dan (3,5iiil;
2,5iiil; 2,0nil; l,5inl lan lugt cho 4 ngay) tiem bap FSH cho bd cho phdi vao budi sang va budi chieu vdi khoang each 12 gid.
Ngay thii' 12 tiem 5inl PGF20 (ddng thdi nit PRID vao budi sang).
Bd ddng due vao ngay thir 14 sau khi tiem. Dugc thu tinh nhan tao bang tinh ddng lanh da dugc giai ddng.
Den ngay thir 7 sau khi phdi gidng tien hanh thu phdi bang phuong phap khdng phau thuat.
Sau khi thu phdi, tiem prostaglandin - F26 cho bd thu phdi. Sau dd 14 ngay tien hanh dat PRID lap lai quy trinh nhu tren lan thir hai,.
Phuo'ng phap thu hoach vd phdn loai loai phoi
Phdi dugc thu hoach bang phuong phap khdng phau thuat vao ngay thir 7 sau khi ddng due va phdi gidng. ngay thir 7 img vdi giai doan phdi dau va phdi nang. Day la giai doan thich hgp cho bao quan va cay truyen. Nhirng phdi chat lugng tdt va phat tnen dung giai doan dugc sir dung trong cdng nghe cay phdi.
KET QUA VA THAO LUAN I
• I
Chimg tdi tien hanh GRTN tren 2 bd bang dung cu chira hormone sinh due Progesterone, FSH va PGF;,, trung binh khoang each lap lai la 35 ngay. Ket qua thu dugc nhu sau:
Lan 1 Lan 2 Lan 3 Lan 4 Lan 5 Lan 6 Lan 7 Lan8 Cdng Mean ± SE
10 12 13 9 10
8 10
9 81 10 ± 1,69
8 9 8 12
8 10
7 8 70 9 ± 1,65
NGUYEN VAN LY - Anh huang gdy rung trung ...
Ket qua kiem tra phiin ling ciia buong triing bo khi GRTN
Bd sau khi GRTN se dugc ihii tinh nhan tao bang tinh cgng ra. Den ngay thir 7 ke tir khi thu linh nhan tao thi lien hanh thu phdi bang phuong phap khdng phau thuat. Trudc khi gigi rira thu hoach phdi, chiing tdi tien hanh khani qua true trang de kiem tra phan img cua budng trirng bd cho phdi. Ket qua the hien d bang 1.
Bang 1. Ket qua theo ddi phan iing ciia budng trung bd khi GRTN
Lan GRTN Sd the vang kiem tra thay ^ Bd sd hieu 552 Bd sd hieu 789 Tdng sd
r8
21 21 21 18 18 17 17 151 19 ± 1,98 Qua Bang 1 cho thay, sau 8 lan GRTN sd the vang binh quan cua hai bd cho phdi la tuong ddi cao bd sd hieu 552 la 10 ± 1,69 dao ddng trong khoang 8 den 13, cdn bd sd hieu 789 la 9 ± 1,56. dao ddng khoang tir 7 den 12. Ket qua cho thay. phan irng ciia budng triing cd sir dao ddng giira cac lan thu phdi.
Ket qua nghien ciai ciia chiing tdi phii hgp vdi ket qua ciia mot sd tac gia khac da cdng bd:
Theo Sacher va cs (1987) nghien ciru GRTN bang PMSG va FSH lap lai lir 4- 23 lan vdi khoang each lap lai la 85 ngay, sd the vang binh quan bien ddng tir 8.7-10,2 cho mgl lan lay phdi. So vdi kit qua nghien ciru trudc day lai Viet Nam. ket qua trong nghien ciru cua chung tdi cao ban ket qua cua tac gia Do Kim Tuyen, (1995) thi nghiem GRTN vdi khoang each lap lai la 85 ngay tren bd lai Fl va F2 giira bd sii'a Ha Lan va bd Laisind cho ket qua sd the vang thu dugc trung binh la 6,3 ± 0,5.
Tuy nhien, so vdi nghien ciru ciia mot sd tac gia khac, ket qua nghien ciru ciia chiing tdi lai thap ban. Tac gia Kim va cs (1987) GRTN bing FSH va PMSG cho bill so the vang khi GRTN bang FSH la 12,3 va sir dung PMSG thi kit qua la 11,9 .
Nhu vay. xet vl mat sd lugng. phan img budng trirng ciia 2 bd dugc GR'FN trong thi nghiem nay la kha tdt; sd lugng thi vang thu dugc kha cao va trong gidi ban dam bao chat lugng tdt.
v l mat thdi gian. trong vdng 9 thang, chiing tdi tiln hanh 8 lan GRTN tren 2 bd. vdi khoang each lap lai trung binh la 35 ngay. So vdi nghien ciru trudc day vdi khoang each lap lai la 85 ngay thi khoang each lap lai giira cac lan GRTN da giam kha nhieu.
Ket qua thu hoach phoi tir bo thi nghiem GRTN
Bd ddng due sau khi GRTN dugc thu tinh nhan tao. ngay thir 7 sau khi thu tinh. chiing tdi tien hanh thu hoach phdi. Sd lugng phdi va le bao trimg thu dugc tir 2 bd thi nghiem qua 8 lan GR TN dugc the hien trong Bang 2.
VIEN C H A N NUOI - Tap chi Khoa hgc Cdng nghe Chdn mid, - So 15 -Thdng 12- 2008
Qua Bang 2 cho thay. sau 8 lan GRTN trung binh sd phdi va sd te bao trirng thu dugc ciia bd sd hieu 552 cao hon so vdi bd sd hieu 789. Bd sd 552 tdng sd phdi va te bao trimg thu dugc qua cac lan GRTN la 57; Sd lugng nay dao ddng tir 5 den 8; trung binh mdi lan thu la 7,13 ± 1,13. 0 bd sd 789, tdng sd phdi va te bao trirng thu dugc la 52, dao ddng tir 5 den 10 va trung binh mdi lan thu la 6,50 ±1,69. Tuy vay. neu tinh rieng sd phdi thu hoach dugc thi ket qua cua 2 bd khdng cd su chenh lech dang ke. 0 bd sd 552 thu dugc trung binh sd phdi la 5,63 ±1,41 so vdi bd sd 789 la 5,38 ±1,69 phdi.
Bang 2. Ket qua thu hoach phdi qua cac lan gay rung trirng nhieu Lan GRTN
Lan 1 Lan 2 Lan 3 Lan 4 Lan 5 Lan 6 Lan 7 Lan 8 Tdng Mean±SE
Sd phdi 5 8 6 7 6 4 4 5 45 5,63 ± 1,41
Sd lugng phdi va te Bd sd hieu 552
Te bao tru'ng 2 0 2 1 2 1 3 1 12 1,50 ±0,93
Tdng : 7 8 8 8 8 5 7 6 57 7,13± 1
sd
,13
bao trirng th
Sd phdi 3 6 7 7 5 7 5 3 43 5,38 ±1,69
u dugc
Bd sd hieu ,789
Te bao trirng Tdng sd 2
1 0 3 1 0 0 2 9 1,13±1,
5 7 7 10
6 7
• • > ' - ^ - 5
5 52 13 6,50 ±1,69 Ket qua ciia chiing tdi phii hgp vdi ket qua ciia mot sd tac gia khac da cdng bd. Theo tac gia Lamberson va cs (1986) tien hanh GRTN lap lai 3 lan sd phdi binh quan ciia cac lan GRTN tuong irng la 5,2; 4,6; 5,2 phdi. Ket qua cua chung tdi cao hon kha nhieu so vdi ket qua nghien ciru cua bd mdn cay truyen phdi Vien Chan nudi nam, (1995). Cac tac gia da nghien ciru GRTN tren bd lai Fl va F2 giira bd sira Ha Lan va bd Laisind cho biet sd phdi thu dugc trung binh la 3,36±0,04 vdi khoang each lap lai la 85-90 ngay. , •. . iMi,
Bang 3. Ty le sd phdi thu dugc so vdi sd the vang kiem tra Lan
Lan 1 Lan 2 Lan 3 Lan 4 Lan 5 Lan 6 Lan 7 Lan 8 Tdng
Sd the vang
10 12 13 9 10
8 10
9 81
B d s d 5 5 2 Sd phdi
5 8 6 7 6 4 4 5 45
Ty le phdi/
the vang (%)
50 66,67 46.15 77,78 60 50 40 55,56 55,56
Sd the vang
8 9 8 12
8' ' 10
. \ . 1 . ' •
8
70
Bd sd 789 Sd phdi
.' :'.
3
. „ . ' • 6 . . .
7
. 1 ':"..
• , ' 5 ' "•'
7
^.•. '<>'ig f b i i .
3 43
Ty le phdi/
the vang
"• (%)
37,5
• 66.67 87,5 58,33
62.5 70
"? 71,43 •"
37,5 . 61,63 Tuy nhien, ket qua cua chiing tdi lai thap ban so vdi nghien cim ciia mot sd tac gia khac. Theo Kim \ a cs (1987) khi GRTN tren bd bang FSH va PMSG cho kit qua, sd phdi thu dugc trong
NGUYEN VAN LY - .4nh huong gdy rung trirng ..
moi lan la 8.2 va 7,3 phdi. Nam 1986, Sergeev NI va cs (1986) nghien cim GRTN lap lai 3 Idn bang PMSG va anti- PMSG cung thu dugc sd phdi tuong img la 6,7; 6,7; 5,8 o moi lin GRTN De danh gia hieu qua cua viec GRTN, chiing tdi da tinh t>' le tao phdi so vdi sd trimg rung.
Ket qua the hien o Bang 3.
Bang 3 cho thay ty le sd phdi tren sd trimg rung thu dugc ciia 2 bd thi nghiem dat t>' le kha cao bd sd 552 dat ty le 55.56% dao ddng trong cac lAn thu phdi tir 40 din 77,78%. cdn bd sd 789 dat ty le la 61.63% dao ddng trong khoang tir 37,5 den 87,5%.
Ket qua cua chung tdi cao ban kha nhieu so \'di ket qua cua Do Kim Tuyen (1995) GRTN tren bd lai Fl va F2 giua bd Ha Lan va bd Laisind t>' le thu phdi tren so trirng rung la 42,08%.
Nhu vay, ty le phdi va trimg thu dugc ciia 2 bd deu cd su bien ddng qua cac lan thu phdi \'a sd phdi thu dugc thap ban so vdi sd trung rung. Dieu nay cd the la do ky thuat thu tinh \'a qua trinh thu phdi chiu anh huong cua cac yeu td nhu: thao tac ciia ky thuat vien trong quo tronh gidi rua. soi tim phdi... Ket qua thu phdi trong nghien cim nay ciia chiing tdi khdng thap hem nhieu so \'di ket qua GRTN ciia nhieu tac gia tren the gidi. Vi vay, cd the ndi quy trinh chiing tdi da su dung kha dam bao ve mat ky thuat va phii hgp \ di dieu kien o Viet Nam.
Ket qua phan loai phoi thu duoc
De dat dugc ket qua tdt khi sir dung hay bao quan phdi, dat ty le cd chira cao cua bd cai nhan phdi sau khi cay, mgl viec lam rat can thiet la phai danh gia phan loai phdi trudc khi quyet dinh sir dung nhu the nao. Trong cdng nghe cay truyen phdi, phdi loai A va B cd the cay ngay cho bd nhan hoac dem ddng lanh de bao quan. Phdi loai C khdng ddng lanh dugc ma chi cd the cay ngay cho con nhan ddng pha, mac dii ket qua cd chua thap ban so vdi khi cay phdi loai A. B. Ket qua danh gia phan loai phdi de sir dimg hgp ly dugc trinh bay trong Bang 4
Bang 4. Ket qua phan loai phdi Lan GRTN
(n = 2
Lan 1 Lan 2 Lan 3 Lan 4 Lan 5 Lan 6 Lan 7 Lan 8 Tdng
bd)
Mean± SE
Tdng Sd
12 15 15 18 14 12 12 11 109 100%
Phdi dii 1 tieu chuan cho cay truyen
(A + B + C)
11
8 14 13 14 11 11 9 8 88 11±2,51
% 66,67 93,33 86,67 77,78 78,58 91,67 75,00 72,73 80,73 80,70
Phdi dii lieu chuan ddng
(A+
11
7 12 12 13 9 9 8 6 76 9,5±2,56
lanh B)
% 58,33 80,00 80,00 72,22 64,29 75,00 66,67 54,55 69,72 69,69
Phdi kem c chain pi
su n 4 1 2 4 3 1 3 3 21 2,63±1.
:hal lugng.
lal trien khdng dur
,19 ig(D)
% 33,33
6,67 13,33 22,22 21,43 8,33 25,00 27,27 19,27 19,29 Qua bang 5 la thSy. sau 8 lan GRTN thu dugc 88 phdi dii lieu chuan diing cay truyen trung binh la 1 1,88 ± 2.51 vdi ty le 80,37%, 76 phdi dii lieu chuan ddng lanh trung binh la 9,50
± 2,56 ty le 69.72% va 21 phdi kem chat lugng. cham phat trien khdng su dung binh quim la 2,63±1,19 vdi ty le 19.27%. Nhu vay, phdi dii tieu chuan diing cho cay truycMi va ddng lanh chiliii ly le kha cao. Ty le sd phdi dii tieu chuan cay va ddng lanh trung binh ciia chiing tdi tuong img vdi ket qua ciia tac gia Nguyen Thi Udc va cs (1996) ty le phdi du tieu chuan
VIEN C H A N N U O I - Tap chi Khoa hgc Cdng nghe Chdn nudi - So 15 -Thangl2- 2008
cay phdi va phdi dii tieu chuan ddng lanh lan lugt la 80,25% va 73,50% vdi kboang each lap la 2,5 thang. Ty le phan tram sd phdi dii tieu chuan ddng lanh ciia chiing tdi cao ban ty le phan tram sd phdi ddng lanh ciia tac gia Hoang Kim Giao va cs (2004) 69,72% so vdi 62,82%. Ty le phdi dii tieu chuan cay ciia chimg tdi thap hon so vdi cac tac gia tren 80,37% so vdi 95,29%. Tuy vay, sd phdi chat lugng tdt, loai A va B ciia chiing tdi cao ban 69.72%o so vdi 65,93%. Nhu vay, vdi 8 lan GRTN tren 2 bd thi nghiem chiing tdi thu dugc 109 phdi va te bao trirng trung binh mdi lan thu la 13,63 ± 0.03 trong dd sd phdi su dung dugc chiem ty le kha cao 80,73% so vdi tdng sd phdi va te bao trirng thu dugc.
KET LUAN
Ket qua gay rung trii'ng nhieu cdn phu thudc vao cac yeu td nhu: sir phan img vdi hormone cua bd cho phdi, dinh duo'ng, ky thuat phdi gidng. qua trinh ddi rira vv... Budc dau ket luan trong dieu kien Viet Nam cd the rut ngan khoang each GRTN tir 2-3 thang xudng 35 ngay, tang sd phdi thu dugc/bd/nam so vdi trudc day.
Dieu nay cd y nghTa vd ciing quan trgng trong chan nudi. Vi nit ngan khoang each giua cac lan khai thac ma van thu dugc ket qua phdi nhieu va tdt ddng nghia voi viec giam chi phi chan nudi. tang hieu qua kinh te, khdng nhiing the. nd cdn cho phep khai thac tan dung nhieu ban tiem nang di truyen ciia nhiing bd cai tdt de dua vao san xuat.
TAI LIEU THAM KHAO
Do Kim Tuyen, (1995). Nghien ciru gay sieu bai noan a bo bang sir dung FSH va prostaglandin F2a.
Luan an PTS KHNN- Ha Noi, 1995.
Hoang Kim Giao, Nguyen Thanh Duong, Do Kim Tuyen, (1996). Ket qua nghien cuu cay truyen phoi bo tai Vien Chan nuoi (1990-1996)
Hoang Kim Giao, Luu Cong Khanh, Nguyen Van Ly. Nguyen Thi Thoa. Phan Le So'n. Dang VQ Hoa, Chu Thj Yen (2004). Gay rung triing nhieu o bo cho phoi.Thong tin Khoa hoc Ky thuat Chan nuoi - So 6 nam 2004.
Kim. H.N, Rovie, R,W. (1987). The use of PMSG antibodies with PMSG for superovulation beef cattle.
Theriogenology Volume 27.
Lamberson W.R. and . Lambeth,V.A (1986). Repeatability of respone to superovulation in Brangus cows.
Theriogenology Volume 26, Issue 5, November 1986.
Nguyen Thi Uoc, (1996). Nghien ciiu gay rung trirng nhieu va gay dong due dong pha trong cay truyen phoi trau bo, Luan an PTS KHNN- Ha Noi.
Sacher, B., H, Niemann, Schilling, E. (1987). Effects of repeated superovulation and embryos recovery and fertility. Animal Breeding- Abstract 55.
Scgeeve, N,l. (1986), The improvement of hormone induction superovulation in donor cows for embryo transfer,Animal Breeding- Abstract 35.
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