DO DAI M I I N DjCH VA THOl GIAN BAO QUAN C O A VACXIN V6 HOAT TU HUYET TRUNG GA DUNG MONTANIDE 50
LAM CHAT BO TRO M I I N DICH
• •
N g u y i n Manh Thing*, N g u y i n N g w Hai ^ Tran Dinh T i i '
TOM TAT
Dd dai miln dich va thdi gian bao quan cua vacxin tu huyd't trung nhii d i u dimg montanide 50 lam cha't bd trp miln dich da dvtdc danh gia, khao sat tren ga thi nghifm khi cdng cudng ddc bang vi khuan Pasteurella multocida chiing Pa va cac chiing vi khudn Pasteurella multocida phan lap tif thuc dia (chiing LD06, BTOl va DH). Kd't qua cho tha'y sau 6 thang tiem vacxin, khi cdng eudng dde b l n g cac chung vi khudn Pasteurella multocida ed ddc life cao nhu chiing Pa, BTOl, ty If bao hd cho ga thi nghifm dat tu: 70% - 80%, brong khi dd toan bd ga dd'i chiing ehdt hdt. Ndu cdng cUdng ddc b l n g chiing vi khudn Pasteurella multocida cd ddc life tha'p hdn nhu chiing LD06, DH, ty If bao hd cho ga thi nghifm dat tuf 80% -100%, tuy nhidn ga ddi chiing vin sdng tif 20% deh 40%. Sau 12 thang bao quan vacxin d nhift dd tif 2''C - 8''C, ty If bao hd eua vacxin cho ga thi nghifm dat 80% khi cdng eudng dde blng vi khudn Pasteurella multocida chung Pa. Nd'u cdng cudng dde b l n g eac chiing vi khudn Pasteurella multocida phan lap tif thifc dia thi vacxin sau 9 thang bao quan d nhift dd tif 2''C - 8''C cd kha nang bao hd cho ga thi nghifm tif 80% ddn 90%.
Tit khoi: Tu huye't trimg ga, Vacxin, Pasteurella multocida, Dd dai miln dich, Thdi gian bao quan, Phan lap, Dde life, Ty If bao hd.
Duration of immunity and storage of fowl cholera vaccine using montanide 50 such as adjuvant
Nguyen Manh Thing , Nguyen Ngoc Hai Tran Dinh Tif
SUMMAEY
The duration of immunity and storage of fowl cholera vaccine using montanide 50 such as adjuvant were tested in chickens by challenged Pasteurella multocida strain Pa and isolated strains from field (LD06, BTOl and DH). The results showed that 6 months after vaccination, chickens were challenged by high virulent Pasteurella multocida strains such as Pa and BTOl, the percentage of protection reachs to 70% - 80% in vaccinated chickens, all controlled chickens died. If chickens were challenged by less virulent Pasteurella multocida strains such as LD06 and DH, then the percentage of protection for vaccinated chickens reachs to 80% 100%, but controled chickens also survive from 20% to 40%. After 12 months store in 2''C - S'C, emulsion vaccine can to protect 80% chickens when challenged against a virulent Pasteurella multocida strain Pa. After 9 months store in 2''C - S^C, emulsion vaccine can to protect chickens from 80%
to 90% when challenged by Pasteurella multocida isolated from field strains such as LD06, BTOl v^DH.
Key words. Fowl cholera. Vaccine, Pasteurella multocida, Duration of immunity. Storage, Isolate, Virulent, Percentage of protection.
' C6ng ty thud'c thii y trung Udng (Navetco),,
^ Khoa chan nu6i thti y-Dai hoc n6ng U m Tp. HCM. 'Hfli thii y Vift Nam.
LDATVANDE
Bfnh tu huye't trung gia e l m do vi khuan Pasteurella multocida gay nen. Tai nude ta, bfnh xay ra tai h l u he't eac tinh thanh, gay nhilu thif t hai v l kinh td cho cae trang trai va hd chan nudi gia dinh (Nguyin Xuan Binh, 1994; LI v a n Tao va cs, 2001; Sa Dinh Chidn, 2001). Mdt trong cac bifn phap lam giam thift hai do bfnh gay ra la tidm vacxin phdng bfnh.
Dd nang cao hifu qua phdng bfnh thi hifu life cua vacxin phai dupe cai tiln. Thay the' chat bd trp phen chua, keo phen b l n g cha't bd trp nhii d i u la mdt hudng di diing de tao ra vacxin cd cha't lupng td't hdn (MeClimon va es, 1994; Islam va es, 2004; Akand va es, 2004).
Chiing tdi da nghidn ciiu su dung dung dich montanide 50 dd chd tao thu nghifm vacxin tu huydt trimg nhii d i u . Vacxin ed hifu life td't ddi vdi ga, tuy nhidn trong thuc td' sU dung, ngoai ye'u td' hifu life thi dd dai miln dich va dd dai bao quan cua vacxin ciing ra't dupe quan tam. Trong thi nghifm nay, chiing tdi khao sat dd dai miln dich va dd dai bao quan ciia vacxin tu huyd't trimg dimg montanide 50 lam cha't bd trp miln dich.
II. NGUYEN VAT L I E U VA P H U d N G PHAP N G H I E N CUtJ
2.1. Nguydn vSt lifu
Vi khudn Pasteurella multocida chung Tg phan lap tif ga bi bfnh THT h Nam bd, thude serotyp A:l.
Vi khudn Pasteurella multocida chiing LD06, BTOl va DH dupe phan lap tif ga bi bfnh THT ci Tiln Giang va Long An, thude serotype A:l.
- Vi khudn Pasteurella multocida chung Pa cd ngudn gdic tif Vifn thii y, dupe dung dd kidm nghifm vacxin tu huyd't trimg gia elm.
Ga khoe manh dupe nudi tif 1 ngay tudi dfh khi lam thi nghifm.
- Vacxin tu huyd't trung vd hoat nhii d i u .
- Cae trang thid't bi eln thidt eho thi nghifm.
2.2. PhUdng phap nghien ciiu
2.2.1. Phuang phip xic dinh dd dai miin dich cua vacxin
Tiem vacxin vd boat nhii d i u cho 10 ga vdi lilu 1,0 ml/con, ga dd'i chiing dupe tidm nude mud'i sinh ly trdn vdi dung dich montanide 50 eung vdi lilu 1,0 ml/eon. Sau khi tidm vacxin 2 tiiang, 4 thang va 6 thang, ga thi nghifm va ga ddi chiing dupe cdng cUdng ddc blng vi khudn Pasteurella multocida chung Pa vdi lilu -300 CFU/eon. Vao thdi didm 6 thang sau khi tidm vacxin, ngoai vifc su dung vi khudn Pasteurella multocida chung Pa de cdng eudng ddc theo quy dinh ciia Cue thu y, chiing tdi edn su dung thdm eac chiing vi khudn Pasteurella multocida phan Ifp tut thifc dia (LD06, BTOl va DH) dd cdng cudng ddc ciing vdi lilu ~300 CFU/eon n h l m danh gia kha nSng bao hd eiia vacxin dd'i vdi cac chiing vi khuan Pasteurella multocida nay.
2.2.2. Phuc/ngphip xic dinh dd dai bio quin cua vacxin
Vacxin dupe bao quan d nhift dd tif 2''C - 8''C. Sau thdi gian bao quan 3 , 6 . 9 va 12 thang, vacxin dupe la'y ra dd kidm tra hifu lUe trdn ga blng each tidm eho 10 ga vdi lilu 1,0 ml/eon, ga dd'i chiing dupe tidm nude mud'i sinh ly trdn vdi dung dich montanide ciing vdi lilu 1,0 ml/con. Vao ngay thii 21 sau khi tidm vacxin, dimg vi khuan Pasteurella multocida chiing Pa cdng eudng ddc vdi lilu -300 CFU/eon. Vao thdi didm 9 thang bao quan vacxin, ngoai vifc su dung vi khuan Pasteurella multocida chiing Pa cdng eUdng dde theo quy dinh ciia Cue thii y, dd danh gia them kha nang bao hd ciia vacxin trdn ga dd'i vdi cae chiing vi khuan Pasteurella multocida phan lap tif thuc dia, chiing tdi sur dung thdm cac chiing vi idiudn LD06, BTOl va DH dd cdng cudng ddc ciing vdi lilu -300 CFU/eon.
I I I . KET QUA N G H I E N CtfU VA THAO LUAN
3.1. D6 dai m i l n dich cua vacxin
Chiing tdi khao sat dd dai miln dich ciia 2
Id vacxin chd' tao thii nghidm. Kd't qua khao miln dich khi cdng eUdng ddc blng vi khudn sat dd dai miln dich ciia vacxin tu huyd't Pasteurella multocida chiing Pa dupe trinh trimg ga dimg montanide 50 lam cha't bd' trp bay trong bang 1.
Bing 1. Hiiu lUc cua vacxin tu huyit triing nhU diu trin gi sau khi tiim 2,4vi6 thing Thai gian
2 thang
4 thang
6 thang
Liiu cong/con -300CFU,
chung Pa
~300CFU, chung Pa
~300CFU, chung Pa
Ld thi nghi$m Vacxin Id 1 Vacxin Id 2 Doi chiing Vacxin Id 1 Vacxin Id 2 Doi chiing Vacxin Id 1 Vacxin Id 2 Ddi chiing
Sogi 10 con 10 con 5 con 10 con 10 con Scon 10 con 10 con 5 con
So song/tong so 9/10 con 8/10 con 0/5 con 9/10 con 9/10 con 0/5 con 7/10 con 8/10 con 0/5 con
Tyl$baoh0(%) 90 80 0 90 90
• 0 70 80 0 Kd't qua thi nghifm cho tha'y, sau 2 thang
tiem vacxin nhii d i u , ty If bao hd eho ga d Id vacxin 1 la 90% va cua Id vacxin 2 la 80%.
Sau 4 thang tidm vacxin, ty If bao hd cho ga d Id vacxin 1 v i n dat 90%, eiia Id vacxin 2 la 90%, cao hdn so vdi ty If bao hd sau 2 thang tidm vacxin (80%), trong khi do toan bd ga dd'i chiing ehdt hdt. Sau 6 thang tidm vacxin, ty If bao hd cho ga d Id vacxin 1 chi dat 70%, ed giam xud'ng so vdi ty If bao hd sau 2 thang va 4 thang tidm vacxin, ty If bao hd eho ga d Id vacxin 2 dat 80%, tha'p hdn so vdi thdi didm sau 4 thang tidm vacxin (90%) nhifng v i n b l n g ty If bao hd sau 2 thang tiem vacxin.
Tai Vift Nam va tren thd gidi, mdt sd' nghidn ciiu v l dd dai m i l n dich ciia vacxin tu huyd't trung gia e l m dimg eac chat bd trp nhii d i u khac nhau ciing da dupe thdng bao. Nam 1994, Ld Lap va cdng su da chd tao vacxin tu huye't trung gia e l m nhii d i u (khdng ndi rd loai n h u d i u ) tif vi khudn Pasteurella multocida chung N41 phan lap tii cac tinh m i l n Trung. Ty If bao hd eho ga sau 4 thang tidm vacxin la 75%, sau 5 thang dat 60%. Ke't qua nghien ciiu eiia Glunder G va es (2004) cho tliS'y sau 6 thang tiem vacxin
tu huyd't trimg nhii d i u (dimg d i u khoang trdn vdi arlacel va tween lam chat bd trp m i l n dich) ty If bao hd eho ga thi nghifm dat 70%. Ty If bao hd ciia cae loai vacxin tu huyd't trung nhii d i u nay tha'p hdn ty If bao hd ciia vacxin vd boat nhii d i u (dung montanide 50 lam chat bd trp miln dich) do chiing tdi chd tao thu nghifm vao ciing thdi didm khao sat (bang 1).
Ngoai chung cdng cudng ddc la vi khuan Pasteurella multocida chiing Pa theo quy dinh ciia Cue thii y, chiing tdi edn sU dung thdm cac chiing vi khuan Pasteurella multocida phan lap tuf thuc dia (LD06, BTOl va DH) dd cdng eUdng ddc ciing vdi lilu -300 CFU/eon n h l m danh gia kha nang bao hd eua vacxin dd'i vdi cac chiing vi khudn Pasteurella multocida nay. Dde life cua cac chiing vi khudn Pasteurella multocida phdn lap tif thuc dia dd'i vdi chudt bach tha'p hdn ddc life ciia vi khuan Pasteurella multocida chiing Pa. LD50 eiia vi khuan Pasteurella multocida chung Pa dd'i vdi eudt bach la 21 CFU/eon, lilu LD50 cua vi khudn Pasteurella multocida ciia cac chiing LD06, BTOl va DH l l n lupt la 124 CFU/eon; 36 CFU/eon va 42 CFU/eon. Kd't qua dupe trinh bay trong bang 2.
Bing2. Hiiu life cua vacxin tu huyit bring vd hoat nhu diu trin gasauS thing tiim vacxin Chung cdng
Pa
Le06
BTOl
BH
Liiu cong/con
~300CFU
~300CFU
~300CFU
~300CFU
Lo thi nghidm Vacxin Id 1 Vacxin Id 2 Doi chiing Vacxin Id 1 Vacxin Id 2 Ddi chiing Vacxin Id 1 Vacxin Id 2 Doi chiing Vacxin Id 1 Vacxin Id 2 Doi chiing
Sogi 10 con 10 con 5 con 10 con 10 con
Scon 10 con 10 con Scon 10 con 10 con S con
So song/
ting so 7/10 con 8/10 con 0/5 con 9/10 con 10/10 con
2/5 con 7/10 con 7/10 con 0/S con 8/10 con 9/10 con 1/5 con
Ty 1$ bao hO (%)
70 80 0 90 100
40 70 70 0 80 90 20 Vao thdi didm 6 thang sau khi tidm vacxin,
khi su dung cac chiing vi khuan Pasteurella multocida phan lap tif thuc dia (LD06, BTOl va DH) cdng cudng dd thi ty If bao hd cho ga cd khac nhau cl cac Id thi nghifm. Dd'i vdi cac chiing cd ddc luc tha'p n h u LD06 hoac DH thi ty If bao hd cho ga dat cao, tif 80% ddn 100%, hiy nhien id dd'i chiing cd tdi 20% hoSc 40%
ga khdng chd't. Trong khi dd, dd'i vdi cac chiing vi khuan Pasteurella multocida ed dde
life cao nhu Pa, BTOl thi ty If bao hd eho ga ed tha'p hdn, ehi dat tif 70% dd'n 80%, tuy nhien toan bd ga dd'i chiing chd't hdt sau khi cdng eUdng dde.
3.2. Khao sat dd dai bao quan ciia vacxin Hifu life miln dich ciia 2 Id vacxin tu huyd't trimg ga dimg montanide 50 lam cha't bd trd miln dich sau mdt thdi gian bao quan d 2''C - 8''C dupe trinli bay trong bang 3.
Bing 3. Hiiu lUc cua vacxin tu huyi't trung vd hoat nhd diu trin ga sau mdt thdi gian bio quin khi cdng bing vi khuin Pasteurella multocida chiing Pa
Thdi gian bio quin
3 thang
6 thang
9 thing
12 thing
Ld thi nghi$m Tiem vacxin Id 1 Tiem vacxin Id 2
Ddi chiing Tiem vacxin Id 1 Tiem vacxin Id 2
Ddi chiing Tiem vacxin Id 1 Tiem vacxin Id 2
Ddi chiing Tiem vacxin Id 1 Tidm vacxin Id 2
Ddi chiing
Sogi 10 con 10 con Scon 10 con 10 con Scon 10 con 10 con Scon 10 con 10 con Scon
Liiu cdng (chung Pa)
~300 CFU
- 300 CFU
~ 300 CFU
- 300 CFU
So song/
tong s6 9/10 con 9/10 con 0/5 con 8/10 con 9/10 con 0/5 con 8/10 con 7/10 con 0/5 con 8/10 con 8/10 con 0/5 con
Ty /# bio /Jd (%)
90 90 0 80 90 0 80 70 0 80 80 0 Sau 3 thang bao quan, ty If bao hd ciia 2 Id
vacxin tu huyd't triing vd boat nhii d i u trdn ga dat 90% khi cdng cUdng ddc blng vi khuan Pasteurella multocida chiing Pa, trong khi dd toan bd g^ d Id ddi chiing chd't hd't. Vao thdi didm 6 thang bao quan, ty If bao hid trdn ga
thi nghifm ciia Id vacxin 1 dat 80%, cd tha'p hdn ty If bao hd ciia Id vacxin 1 sau khi bao quan 3 thang, ty If bao hd eua Id vacxin 2 vin dat 90%. Sau 12 diang bao quan, ty If bao hd trdn ga thi nghifm eiia Id vacxin 1 giu nguydn la 80%, khdng thay ddi so vdi ty If bao hd sau
6 thang va 9 thang bao quan. Ty Id bao hd trdn ga thi nghifm ciia Id vacxin 2 sau 12 thang bao quan dat 80%, ed giam so vdi ty If 90% sau 6 fliang bao quan. Tuy nhidn vao thdi diem 9 thang sau khi bao quan, ty If bao hd tren ga thi nghifm eua Id vacxin 2 chi dat 70%, tha'p hdn so vdi thdi didm bao quan sau 12 thang (80%).
Vao thdi diem sau 9 thang bao quan vacxin.
chiing tdi ed su dung thdm cac chung vi khuan Pasteurella multocida phan lap tif thuc dia (LD06, BTOl va DH) dd cdng eudng ddc nham kidm tra lai kha nang bao hd eiia vacxin dd'i vdi cae chiing vi khuan nay trdn ga va do mdt sd khd khan ciing n h u d i i u kifn kinh phi, chiing tdi chi sU dung Id vacxin 2 dd kidm ha.
Kd't qua dupe trinh bay trong bang 4.
Bing 4: Hiiu luc ciia vacxin tu huyi't triing vd hoat nhu diu trin ga sau khi bio quin 9 thing Chung cong
Pa LD06
BTOl DH
Lieu cong/con
~300CFU
~300CFU
~300CFU
~300CFU
Lo thi nghidm Tiem vacxin 16 2
Doi chiimg Tiem vacxin 16 2
D6i chumg Tiem vacxin 16 2
Doi chung Tiem vacxin 16 2
Doi chimg
S6ga 10 con 5 con 10 con
5 con 10 con
5 con 10 con
5 con
So song/ tong so 7/10 con
0/5 con 9/10 con
1/5 con 8/10 con
0/5 con 9/10 con 1/5 con
Ty I? bao ho (%) 70
0 90 20 80 0 90 20 Khi su dung cae chiing vi khudn Pasteurella
multocida pMn lap tif thvfe dia (LD06, BTOl va DH) de cdng cudng ddc sau khi tidm vacxin da dupe bao quan 9 thang thi ty If bao hd eho ga khdng gi£5ig nhau d eac Id thi nghifm. Ty If nay dao ddng tif 70% d ^ 90% tuy thudc vao chung cdng cudng dde. Khi dung cac chiing vi khuan Pasteurella multocida cd ddc life cao nhu chiing Pa va BTOl dd cdng thi ty If bao hd cho ga lln lupt la 70% va 80% nhung toan bd ga dd'i chiing chd't he't. Trong khi dd, ne'u sU dung eac chiing vi khuan Pasteurella multocida cd dde life thap hdn la LD06 va dd cdng cudng dde thi ty If bao hd cho ga thi nghifm ldn tdi 90%, tuy nhidn v i n edn 20%
ga d Id dd'i chiing khdng chd't.
Theo quy dinh cua Cue thii y, vacxin tu huydt trimg gia d m dupe coi la dat tieu e h u ^ v l hifu luc ndu ty If bao hd cl gia d m thi nghifm dat tif 50% hci len. Nhu vay, kit qua khao sat cua diiing tdi eho tha'y khi bao quan 12 thang d nhift dd tit 2°C d6i ffC, vacxin tu huydt trimg vd hoat nhii diu (dung montanide 50 lam cha't bd trp miln dich) do chiing tdi chd'tao thuf nghifm v i n edn kha nang bao hd cho ga, dat tieu chuan theo nhu quy dinh ciia Cue thii y.
IV. KET LUAN
Vacxin tu huye't triing vd boat nhu d i u (dimg montanide 50 lam cha't bd' trp miln dich) do chiing tdi chd tao thu nghifm cd dd dai miln dich dd'i vdi ga thi nghifm la 6 thang va dd dai bao quan d nhift dd hf 2''C - 8''C keo dai tdi 12 thang.
TAI LIEU THAM KHAO
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2. Sa Dinh Chi&\, 2001. Nghi§n ciiu Wnh tu huyfi't trung ol San La, mdt s6' d3c tinh cua Pasteurella multocida phSn ISp dudc, bifn phap phong tri. Lu|n an ti€h sT ndng nghifp. Ha N6i.
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4. Quy trinh ky thuat ki^m nghifm vacxin dung trong thii y, 1994. Nha xua't ban Ndng nghifp - Bd Ndng nghifp va CNTP.
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8. Islam M.A., Samad M.A., Rahman M.B., 2004.
Evaluation of alum precipitated formalin killed cholera vaccines with their immunologic responses in ducks. Inter. Jour, of Sci. Infer., 3(2). Pages: 140 -143.
9. MeClimon L.B., Click B., Dick J.W., 1994. Effect of three commercially available adjuvants on the production of antibodies to Pasteurella mutocida in broilers. Avian Dis., 38 (2). Pages: 3 5 4 - 3 6 1 .