KHOA HOC KY THUAT THU Y - TAP XVIII - SO 5 - 2011
DANH GIA CHAT LUpNG COA VACXIN DA GIA PHONG BA BENH NIUCATXON, GUMBORO, VIEM PHE QUAN
TRUYEN N H I I M 6 GA OUOC SAN XUAT THCT NGHIEM TAI XI NGHIEP THUOC THU Y TRUNG l/ONG
Nguyfin Hdng Minh^ Trln Thj Lien', Trtfdng Quangf Bill Thi Ha', Bui Thi Nga My' T6M TAT
Thi nghifim danh gia cha't Itfdng ctia 5 Id vacxin da gia phdng ba bfinli Niucatxdn, Gumboro va vifim phfi' quan bruyin nhilm (IB) d ga san xudt thii nghifim tai Xi nghifip thud'c thti y hrung tfdng , kfi't qua cho tha'y vacxm dat tifiu chudn vd triing, an toan va hifiu lire.
Bdng phtfdng phap cdng ctfdng ddc, xac djnli dtfdc ty 16 bao hd ciia ga dd'i vdi bfinh Niucatxdn sau khi dtfdc sii dung vacxm da gia la 86,66%.
7 ngay sau khi nlid vacxin da gia idn 2, hifiu gia kliang tlid Niucatxdn trong huyfi't thanh ctia ga bat ddu tang (3,25 ± 1,22 log2), dat cao nliS't luc ga 35 ngay tudi (6,35 ± 0,50 log2), sau do giam ddn d ^ 49 ngay tudi cdn 3,15 ± 0,75 log2.
Tiyjdioa: Ga, Vacxin, Bfinh Niucatxdn, Bfinli Gumboro, Vifim phe'quan truyin nhilm, San xua't thu nghifim, XNTTYTW
Quality evaluation of a tri-valent vaccine (against Newcastle disease, Gumboro and Infectious Bronchitis) produced in the
Central Enterprise for Veterinary Drugs
Nguyen Hong Minh , Tran Thi Lien , Truong Quang BiJi Thi H^, Bui Thj Nga My SUMMARY
Assays for evaluation of tiie quality of tri-valent vaccine for chickens against Newcastle Disease (ND), Gumboro and Infectious Bronchitis (IB) produced in the Central Enterprise for Veterinary Drugs showed that the vaccine met standard requirements in sterility, safety and efficacy.
The efficacy of the vaccine agamst ND was confirmed by an 86.66% of survival of the vaccinated chickens after the virulent challenge. The antibody titer against ND began to raise (3.25 ± 1.22 log2) at day 7 after the second vaccination, culminated at the chicken age of 35 days old (6.35 ± 0.50 Iog2), then decreased to 3.15 ± 0.75 Iog2 at 49 days old.
Key words. Chicken, Vaccine, Neweastie disease, Gumboro, Infectious bronchitis, PUot production 1. DAT VAN DE ra't Idn. Bori Ie bfinh Gumboro gay suy giam
Chan nudi ga tap trung, sd' ltfdng Idn, nfi'u miln dich, tao dilu kifin cho cac bfinh khac bfinh bruyin nhifim, dSc bifit la ba bfinh phat sinh. Bfinh Niucabtdn va vifim phfi' quan Niucatxdn, Gumboro va vifim phfi' quan truyin nhifim xay ra cl ga moi Itfa tudi, ty lfi bj b-uyin nhifim xay ra thi tdn tha't chdc chdn se bfinh ra't cao. Rifing vdi bfinh Niucatxdn, ngay
' Xi nghi¥p lhu6c lliu y TU' 'Truiing D?i hgc Nong nglii?p - H& Npi
ca khi can thifip bdng khang thfi ddc hifiu ciang khdng thd khd'ng chfi' dtfi?c ty lfi chfi't. VI thfi' bifin phap khdng thd thifi'u dtf^c la stf dung vacxin phdng ba bfinh trfin cho ga. Hifin tai da cd cae vacxin ddn gia va khdng nhilu vacxin da gia phdng ba bfinh trfin. Tuy nhifin vdi vacxin ddn gia thi bat l^i Idn nha't la trong chtfdng trinh phdng bfinh, ngtfdi chAn nudi phai nhilu Idn bdt ga cho ud'ng hoac tifim vacxin,
Dd gdp phdn khdc phuc tdn tai trfin, Xi nghifip thudc thii y trung tfdng (XNTTYTU) dang san xua't thii nghifim vacxin da gta phdng Cling llic ba bfinh Niucatxdn, Gumboro va vifim phfi' quan truyin nhilm.
Dfi dam bao cha't ltfdng ciia san pha'm trtfdc khi dtfa ra thj trtfdng, chiing tdi tifi'n hanh kifim tra cae chi tifiu cha't Itfdng ciia vacxin da gia va nghifin ctfu dap ling miln djch chd'ng bfinh Niucatxdn ciia ga dtfdc sii dung vacxin da gia san xua't tai Xi nghifip thud'c thii y trung tfdng.
II. NOI DUNG, NGUYEN LIEU VA PHU'ONG PHAP NGHIEN CUtl 2.1. Ndi dung nghifin ciiru
- Danh gia cac chi tifiu vd triing, an toan, hifiu Itfc ciia vacxin da gia phdng ba bfinh Niucatxdn, Gumboro va vifim phfi' quan truyin nhilm d ga
- Xac dinh ty lfi bao hd ciia ga dd'i vdi bfinh Niucatxdn dtfdc sii dung vacxin da gia bang phtfdng phap edng ctfdng ddc
- Khao sat sir bifin ddng hifiu gia khang thd Niucatxdn trong huyfi't thanh ciia ga sau khi stf dung vacxin da gia.
2.2. Nguyfin lifiu nghifin cihi
+ Vacxin Niucatxdn nhtfdc ddc ddng khd chung Lasota dtfdc san xua't tai Xi nghifip tfiudc thii y trung tfdng.
+ Vacxin da gia rJurcic ddc ddng khd phdng ba bfinh Gumboro, Niucatxdn, vifim phfi'quan truyin nhilm ciia ga dtfdc san xua't thii nghifim tai XNTTYTU
+ Chiing virut ctfdng ddc Niucabcdn VN91 dtfdc ca'p bdi Trung tam kidm nghifim thudc thii y Trung tfdng I.
+ Ga thjt thtfdng pha'm 1 ngay tudi dutJc mua tir Trai ga triing gid'ng Ba Vi va nudi tai hai thtfc nghifim ciia XNTTYTLT dd sii dung phii h^p cho t ^ g thi nghifim. Toan bd khu vtfc chan nudi dtfdc kidm soat vl mat an toan sinh hpc, ngudn ntfdc udng va thiic an dua vao.
2.3. Phtfdng phap nghifin cii:u
- Kifi'm h-a vd trung: Tifi'n hanh theo tifiu chudn nganh TCN 161 — 92.
- Kifi'm tra an toan: Mdi Id kidm tra 20 ga (dat tifiu chuan thi nghifim), mdi con sii dung 10 lilu vacxin theo dtfdng nhd mdt, nhd miii.
Theo ddi ga trong 14 ngay
- Kidm tra hifiu Itfc: 20 ga 7 ngay hidi (dat tifiu chudn thf nghifim) dtfdc nhd mdt, nhd miii vacxin da gia 2 Idn liic 7 va 14 ngay tudi, mdi Idn 1 lilu vacxin, 10 ga cimg liia tudi va ngudn gdc, khdng sii dung vacxin lam ddi chtfng. 14 ngay sau khi sii dung vacxin idn 2, la'y mau ga d cac Id thi nghifim, chdt huyfit thanh va danh gia hifiu lire cae Id vacxin da gia thdng qua vific xac djnh khang tfid Niucatxcm, Gumboro va vifim phfi' quan truyin nhilm.
+ Kifim tra khang thd Niucatxdn bdng phan tfng ngan b-d ngtfng kfi't hdng cdu (HI) (Nguyin Nhtf Thanh va cs, 2001).
+ Kifi'm tra khang thd Gumboro bdng phan ling ELISA: Su: dung bd kit ELISA virut Gumboro thtfdng mai cua hang IDEXX, My.
Danli gia kfi't qua phan ling: NhQhg mlu huyfi't thanh cd ty 16 S/P < 0,2 dtfdc coi la am tinh, ty lfi S/P > 0,2 dtfdc coi la dtfdng tinh
+ Kifim tra khang thd vifim phfi' quan b-uyIn nhilm (IB) bdng phan ling ELISA: Su"
dung bd kit ELISA virut IB ciia tap doan Synbiotics.
Danh gia kfi't qua phan ling: Nhiing mdu huyfi't thanh cd ty lfi S/P £ 0,15 dtfdc coi la am tinh, ty lfi S/P > 0,15 dtfdc coi la dtfdng tinh.
- Danh gia hifiu Itfc vacxin Niucatxdn bdng cdng Ctfdng ddc
45 ga 7 ngay tudi dtfdc cliia thanh 3 Id, mdi Id 15 con. Ld 1 - sii dung vacxin ddn gia. Id 2 - sii dung vacxin da gia. Id 3 - khdng su: dung
KHOA HOC KY THUAT THU Y - T A P XVIII - s 6 5 - 2011 vacxin (dd'i chiing). 14 ngay sau khi sii dtjng
vacxin Idn 2, ga cac Id thi nghifim dtfdc cdng ctfdng ddc bang chiing virut Niucatxdn VN91 vdi lilu lO^EIDso/ 0,25 ml theo dtfdng tifim bSp, Theo ddi cac bifiu hifin trifiu chiing lam sang ciia bfinh Niucatxdn d" cac Id ga thi nghifim lifin tuc 14 ngay.
- Cac kfi't qua thi nghifim dtfdc tinh toan bdng chtfdng trinh Microsoft Excel
III. KET QUA NGHIEN CUtJ VA THAO LUAN
3.1. Kfi't qua danh gia cac chi tifiu chS't ltfdng ciia cac Id vacxin da gia
Cac Id vacxLi san xua't thii nghifim dtfdc la'y mdu tlieo TCN 160 — 92 "Vacxin thii y — Quy trinh la'y mdu va sii dung mdu trong kifi'm nghifim".
3.1.1. Kit qua kiim tra v6 triing.
Kfi't qua kifim tra dtfdc trinh bay d bang 1.
Bangl. Kit qua kiim tra vd trung cac Id vacxin da gii
Ld vacxin
1 2 3 4 5
Th&i gian theo ddi
(ngdy) 7 7 7 7 7
Loai mdi trucmg kidm tra Thach mdu
(2 dng)
-
Thach ndm (2 dng)
~
Nude thit (2 dng)
NudK thit gan ydm khi (2 dng)
Kdt tudn
Bat Bgt est Dgt Dat Ghi ihn: •• dm linh (khdng co \i khudn mgc)
Tit bang 1 cho tha'y, cac 16 vacxin san xua't thii nghifim dtfdc kidm tra trfin 4 loai mdi trtfdng, mdi loai 2 d'ng. Sau 7 ngay theo doi diu cho kfi't qua am tinh, khdng cd vi khuSn moc. Can cii tiieo 10 TCN 161 — 92, cac Id
vacxin da gia diu dat kfi't qua kifim tra theo tifiu chuan nganh.
3.1,2. Kit qua kiim tra an toan Kfi't qua dtfdc trinh bay d bang 2.
Bang 2. Kit qua kiim tra an toin cac 16 vacxin da gia
LdTN 1 2 3 4 5
Sd gi TN (con)
20 20 20 20 20
iJdu vacxin sCrdung (tidu)
10 10 10 10 10
Dudng dtfa vacxin
Nho mat, nho mui
Thdi gian theo ddi
14 ngdy Kdtqud S&n'a 20/20 20(20 20/20 20/20 20/20
Chdt 0/20 0/20 0/20 0/20 0/20
Ke!
tudn Dat Dat Dat Dat Dat
lit kfi't qua d bang 2 cho tha'y: Mac dti ga dtfdcsii dung vacxin ga'p 10 idn binh thtfdng, nhtfng sau 14 ngay theo doi, ta't ed sd' ga diu s ^ g khoe, phat tridn binh thtfdng.
Nhtf vay, cae Id vacxin da gia san xua't thii nghifim dat tifiu chudn an toan.
3.1.3. Kitquakiim trahiiultic Cae Id vacxin da gia dtfdc kifim tra hifiu Itfc bdng each xac djnli hifiu gia khang the trong huyfi't thanh eiia ga sau khi stf dung vacxin.
Kfi't qua dtfdc trinh bay d bang 3.
Bang 3. Kd't qua kid'm tra liiSu luc cac Id vacxin da gia
Ld TN
1 2 3 4 5
Khing thd Gumbon>
Ty Id S/P tmng binh MD
(n'20) 0,46±0,17 0,58±0.28 0,53±0,23 0.48±0,19 0,51±0,22
BC (n'lO) 0,16±0,15 0,19±0.09 0,11±0,03 0.15±0.06 0,12±0,08
Khdng thd IB Ty id S/P tmng binh MD
(n'20) 0,23±0,07 0,21±0.09 0,23±0,09 0.34±0,12 0,31±0,11
BC (n'10) 0,11±0,07 0,12±0,03 0,10±0,09 0.04±0,03 0,14±0,09
Khdng thd Niucafxon (log2) Hidu gid khing thd tmng binh
MD (n=20) 3,86±1,47 3,46t1,22 3,38t1.15 3,58*1.46 4.24±1,08
BC (n=10) 2,17±1,14 1.77i0,63 1,50±1,01 1.83±1.23 0.97±0,89 Ghi (Iui: MD - Micii dich
DC- Ddi chimg
Tir kfit qua bang 3 cho tha'y, khang thd Gumboro cd ty lfi S/P trung binh cua Id ga miln dich diu Idn hdn 0,2 (bifi'n ddng tir 0,46
— 0,58) va Id ga dd'i chiing nhd hdn 0,2 (bifi'n ddng tir 0,11 — 0,19). Theo htfdng ddn cua hang IDEXX, My san xua't bd kit ELISA virut Gumboro thi vacxin da gia dat chi tffiu hifiu Itfc chd'ng bfinh Gumboro.
Khang thfi viem phfi'quan truyin nhilm, ty lfi S/P trung binh cua ga d Id miln djch diu Idn hdn 0,15 (bifin ddng zit 0,21 — 0,34), ciia ga d Id dd'i chiing diu nhd hem 0,15 (bifi'n ddng tli 0,04 — 0,14). Theo htfdng ddn ciia hang Synbiotics san xua't bd kit ELISA thi-cac
Id vacxin da gia dat chi tifiu hifiu Itfc chAig bfinh vifim phfi'quan truyin nhilm.
Hifiu gia khang thd Niucatxdn trung binh cua ga d Id miln djch diu Idn hdn 3 log2 (bifi'n ddng tir 3,38 — 4,24 log2) va ciia ga d Id ddi chtfng diu nhd hdn 3 log2 (bifin ddng til 0,97
~ 2,17 log2). Tham chifi'u Iheo tifiu chu^
ASEAN va DIE khang dinh cac Id vacxin da gia diu dat chi tifiu hifiu Itfc chd'ng bfinh Niucatxdn.
3.2. Ket qua cdng ctfdng ddc ga sau khi sii dung vacxin Niucatxdn ddn gia va vacxin da g "
Tifi'n hanh bd' tri thi nghifim nhtf phdn phuong phap da md ta. Kfit qua dtfdc trinh bay d bang 4.
Bang 4. KS't qua cdng cucfng dSc gi cic 16 thi nghiSm Cic chi tieu theo ddi
Tu6i gd nho vacxin Idn ddu (ngdy) Dufdng d\sa vacxin Tuoi gd nho vacxin Idn 2 (ngdy) Tu6i gd c6ng cu-dyng d j c (ngdy) So gd cdng cu'&ng ddc (con) Lieu cdng cudng ddc Olfdng cdng cu-dng ddc So gd sau Vtv cdng cudng ddc C6 tri$u
Chung Idm sdng vd chdt (con) So gd sdng sau khi cdng Cudng ddc (con) Ty Id bao hd (%)
Ldi 1 Nhd mdt, mui
14 28 15
I.S2 7 Nho mdt, mOi
14 28 16
1.6 3
•
28 15 10'EID5o'0.25ml Ti&m bdp
1
15 93,33
TiSm bdp 2
15 86.67
Tiem bdp 15
0 0
20
KHOA HOC KY THUAT THU Y - TAP XVIII - s 6 5 - 2011 Qua kfi't qua d bang 4 cho tha'y. Id 1 (sii
dung vacxin ddn gia) cd 1 con. Id 2 (sii dung vacxin da gia) cd 2 con cd trifiu chting ISm sang va chfi't. Ty lfi bao hd ciia ga Id 1 la 93,33% va Id 2 la 86,67%. Trong khi do Id 3 (khdng sii dung vacxin) toan bd ga cd trifiu chiing lam sang va chfi't trong thdi gian theo ddi. Tham chifi'u tifiu chudn nganh TCN 181
— 93 "Quy trinh kifim nghifim vacxin Niucatxdn chiing Lasota", ga dtfdc sii dung vacxin dcfti gia va da gia phdng bfinh Niucabtdn san xua't tai XNTTYTU dat ehi tifiu hifiu Itfc.
3.3. K^t qua xac djnh hifiu gia khang thfi' Niucatxdn trong huy^t thanh cua ga sau khi sii dung vacxin da gia
150 ga dtfdc chia thanh 2 Id. Ld I -100 con (Id miln dich), nhd vao mdt, mui mdi ga 1 lilu vacxin da gia liic 7 va 14 ngay tudi. Ld II — 50 con, khdng sii dung vacxin (Id ddi chiing).
Ga cac Id thi nghifim dtfdc la'y mau, chdt huyfi't thanh vao liic 7, 14, 21, 28, 35, 42 va 49 ngay tudi. Mdi Idn la'y nglu nhifin. Id I - 10 mdu, Id II - 5 mdu. Cac mdu huyfi't thanh ciia ga dtfdc xac djnh hifiu gia khang thd Niucatxdn bdng phan ting HI . Kfi't qua dtf^c trinh bay d bang 5 va dd thi 1.
Bang 5. Kit qua xic dinh hiiu gia khing QiiNiucatxdn trong huyit thanh cua gi bing phan ting HI
Ldthi nghi$m
1 11
Dung luvng m§u
10 5
Hi$u glA khdng thi trung binh (tog2) 7 ngdy
2,72 ±1,15 2,85 ± 0.67
14 ngdy 3,25 ±1,22 2.17 ±1,01
21 ngay 5.13 ±0,81 1,8 ±1.41
28 ngay 5,23 ±1,30 1,55 ± 0,89
35 ngay 6,35 ±0,50
0
42 ngay 5,08 ±1,46
0
49 ngdy 3,15 ±0,75 0
Tli sd' lifiu trong bang 5 cho tha'y: Tai thdi didm 7 ngay tudi, hifiu gia khang thd thu ddng Niucatxdn trung binh ciia ga la 2,72 ± 1,15 log2 (Id I) va 2,85 ± 0,67 Iog2 (Id II), diu nhd hdn 31og2. Do vSy, sur dung vacxin Niucatxdn vao thdi didm nay la phii hdp, khdng anh htfdng d m dap ling miln dich chii ddng.
Ld 1, ga sau khi sii dung vacxin Idn 1, hifiu gia khang thd Niucatxdn bdt ddu tSng nhe (3,25 ± 1,22 log2) d 14 ngay tudi, nhtfng 7 ngay sau khi tai chiing vacxin Idn 2, hifiu gia khang thd tang cao 5,13 ± 0,81 log2 va dat dmh cao d 35 ngay tudi (21 ngay sau khi sii dung vacxin
Idn 2) 6,35 ± 0,50 log2. Sau dd hifiu gia khang thd giam ddn, dfi'n 49 ngay tudi chi cdn 3,15 + 0,75 log2. Trong khi dd. Id thi nghifim II — ga khdng dtfdc sii dung vacxin, hifiu gia khang thfi thu ddng giam ddn va khdng phat hifin dtfdc khang thd d 35 ngay tudi.
Nhtf vay, ga dtfdc sii dung 2 Idn vacxin da gia llic 7 va 14 ngay tud'i dtfdc bao hd dd'i vdi bfinh Niucatxdn trfin 7 hidn tudi. Bifi'n ddng hifiu gia khang thd Niucatxdn trong huyfi't thanh ciia ga cac Id thi nghifim dtfdc thd hifin ti-fin dd thi 1.
7
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Ol
5 5
C
i 3
•IB
^ 1 0
Hl$u gid khdng thi Newcastle trung bInh
y^
/ \
^ \
• • < - . , _ ^
~~~*~~~-X
\ _
7 14 21 28 35 42 49 Ngayluil
- ^ L S M D - ^ L d D C
Dithil. Biin d6ng hiiu gia khing thS Niucatjxfn trong huyit thanh dia ^ sau dt^ sii dung vacxin
I V . K E T L U A N
Cac Id vacxin d a gia p h d n g b a bfinh Niucatxdn, G u m b o r o v a vifim phfi' q u a n t r u y i n nhifim d g a s a n xua't t a i X N T n i T U d a t tifiu c h u a n v d t r u n g , a n t o a n v a hifiu Itfc.
Bdng phtfdng p h a p c d n g ctfdng d d c c h i i n g v i r u t N i u c a t x d n V N 9 1 v d i l i l u lO^EIDa,/ 0,25 m l t h e o d t f d n g tifim b d p , ty lfi b a o h d g a dd'i v d i bfinh N i u c a t x d n s a u k h i dtfdc sii d u n g vacxin d a gia Id 86,66%.
7 n g a y s a u k h i n h d vacxin d a g i a Idn 2, hifiu gia k h a n g t h d N i u c a t x d n t r o n g huyfi't I h a n h ciia g a b d t d d u t i n g ( t r u n g b i n h 3,25 log2), d a t cao nha't luc g a 35 n g a y tudi ( t r u n g H n h 6,35 log2) v a g i a m d d n , dfin 49 n g a y tudi v d n c d n trfin n g t f d n g b a o h d q u y djnh ( t r u n g b i n h 3,15 log2).
T A I L I E U T H A M K H A O 1. Cue thu y, 1994. Quy trinh ky thuSt kidm n ^ f i m
vacxin diing trong thu y. NXB NOng nghifip Ha NOi.
2. Phan Van Luc, N g u y i n Thiiy Hdng, Lai Kim Lan, ViJ Thi Chinh (1996), "Mdi tUdng quan giia ham ltfdng khang thd Itfu hanh va siic bao h6 chdng virut ctfdng d6c Niucatxdn", Tuydn tip c6ng trinh nghifin cuu KHKT gia cdm 1986 - 1996, NXBNN Ha NOi, tr. 216 - 219.
3. N g u y i n Nhtf Thanh va cOng stf, 2001. Vi sinh vit t h u y . NXBNN Ha N0i
1. ASEAN Cooperation in Food, Agriculture and Forestry. Asean standards (or animal vaccines.
Second Edition, Livestock publication series No.2A.
5 OIE, 2008. Manual of diagnostic tests and vaccines for terrestrial animal (mammals, birds and bees). Sixth Edition.
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