Nguyin Thi Kim Lan vd Dtg Tap chi KHOA HOC & CONG NGHE 164(04): 8 7 - 9 5
XAC DINH SEROTYPE, B ^ C LV'C VA TINH KHANG KHANG SINH CUA 3 LOAI VI KHUAN GAY VIEM PHOI 6 LON TAI TINH BAG NINH
Nguyen Th) Kim Lan , Nguyen Thi NgSn, Nguyin Van Quang, Phan Thj Hing Phuc, Le Minh, Ph^m Di|u Thuy, Tran Niiat Thing, Duong Thj Horig DuySn Trudng Dai hgc Nong Lam - DH Thai Nguyen T O M T A T
Ket qua xac dinh Serotype, dpc lye va tinh man cam khang sinh ciia ba loai vi khuan gay viSm ph6i d Ipn tai tinh B5c Ninh cho thiy: Cac chiing vi khuln Actinobacillus pleuropneumoniae {A.
pleuropneumoniae) phan lap dupe thu^c serotype 2 (63,63%) va serotype 5 (36,36%); cac chiing vi khuln Pasteurella multocida {P multocidd) phan \%p dupe thupc serotype A (50,00%) va serotype D (50,00%); cac chung vi khuln Streptococcus suis (S. stiis) phan lap dupe thu^c serotype 2 (41,18%), serotype 7(29,41%) v^ serotype 9 (29,41%). Cac chiing vi khuln phSn l^p dupe diu CO doc luc cao, gay chit 95 - 100% s l chuQt thi nghifm sau khi c5ng cudng d6c 1 2 - 3 6 gid. Cac chiing vi khuan A. pleuropneumoniae, P. multocida va S. suis phan lap dupe deu man cam manh vdi khang sinh Ceftiofiir, amoxicillin, Florfenicol.
Tii khoa: serotype, dgc luc, khdng khdng sinh, vi khudn, Ifm
D A T V A N
DE
Bac Ninh li mdt tinh cd nghe chan nudi Ipn phat triln. Theo thdng ke Cue thing ke chan nudi tinh den 01/10/2016, ting dan Ipn cua tinh la 418.278 con, trong dd 371.219 Ipn thjt, 46,292 lan nai va 767 Ipn dye giong. Ciing nhu nhieu dja phuang khic trong ca nude, chan nudi Ipn tap trung theo quy md Idn d tinh Bac Ninh da va dang gap rat nhieu kho khan, dac biet la dich benh, anh hudng Idn tdi nang suat chin nudi. Trong nhiing nam 2007 - 2013, hpi chirng rdi lo^n sinh sin va hd hap (PRRS) da bung phat manh tren dan Ion ciia tinh Bac Ninh, gay thiet h^i Idn ve kinh te.
Lpn mic benh chit chii ylu trong tinh trang viem phii - mang phii rit nang do cae vi khuan gay viem phdi ke phat khi sire mien dich cua lpn suy giam.
Mac du trong nam 2014 - 2015 dich PRRS khdng xay ra tren dja bin tinh BIC Ninh, song qua kit qua giam dinh sy luu hanh virus, chung tdi thay ti le lpn cd huyet thanh duang tinh vdi virus PRRS kha phd biln (21,63%), ddng nghTa vdi nguy co cao dich PRRS cd the biing phat trd lai.
Tel: 0912 660 317. Email- [email protected] vi
Thuc te tr€n cho thiy, neu dich PRRS xay ra, van de rit quan trpng la phai khong che dupe cac vi khuan gay viem phoi kl phat, tir dd se han che dupe ti le Ipn chet, giam thieu dupe tac hai ciia dich PRRS tren dan lpn.
De dat dupe myc dich trSn, trong nam 2015 - 2016, chiing toi da phan lap 3 loai vi khuan A.
pleuropneumoniae, P. multocida va S. suis, xac dinh serotype, dpc lye vi tinh man cam khang sinh cua chiing de xay dung phac do dieu trj viem phoi hieu qua cao cho lpn VAT LIEU, N O I D U N G VA PHUONG
P H A P NGHIEN CUUVat lieu
* Mdu b^nh pham
Mau phdi va cudng hong cua lpn: Sau khi ed ket qui xet nghiem huyet thanh duang tinh lay tat ca nhiing mau phoi vi cudng hong ciia nhiing lpn duang tinh va nhiing mau phdi va cuong hpng cua nhiing lpn am tinh nhung co bieu hien viem phdi de ed du 300 mau dung phan lap vi khudn A. pleuropneumoniae, P.
multocida vd S. suis.
* Dong vat thi nghiem
Chupt nhat trang khoi lupng 1 8 - 2 0 gam, khde manh.
87
Nguyen Thi Kim Lan vd Dtg I ap em KJ^Urt nv'-' «• v-uiiu
* Cdc logi hod chdt, moi tnecmg va nguyen vat lieu khdc
- Cac loai moi trudng diing de nudi cay, phan lap vi khuln do hang Oxoid (Anh) va Merck (Phap) sin xuit nhu mdi trudng nude thit, thach thudng, thach mau, th^ch MaeConkey, thach Chocolate,...
- Moi trudng phSn lap vi khuin va tang sinh nhu thach BHI cd bd sung 5% mau cim hoac mau bd; thach chocolate; thach TSA (Tryptone Soya Agar) co bo sung 1 - 3%
fresh Yeast Extract; TSB (Tryptone Soya Broth) cd bd sung 1- 3% fresh Yeast Extract va 5%. huyet thanh ngya; nude thit TYE (Tryptone Yeast Extract Broth); thach dmh dudng PPLO cd bo sung 0,1% glucose, 8- 10% YE tuoi va 5%i huyet thanh ngya;
- Mdi trudng xac dinh cac dac tinh sinh hda:
Oxidase, Catalase, Indol, Urerase, O.N.P.G.
Cac lo^i dudng nhu glucose, mannitol, trehalose, arabinose, lactose, raffinose, maltose,...
- Cac vat lieu hda chat khac nhu giay thd phan ung Oxidase, dung dich H202 3%i, nude mudi 6,5%, thudc thu Kovac's, giay tam khang sinh, huyet thanh ngya, thiamine,...
- Cac loai hoa chat, moi trudng dung cho cac phan ling ngung ket, huyet thanh hpc va phan ling PCR dupe chuan hda theo quy trinh nghien cuu cua Bp mon Vi triing, Bd mon Virus, Vien Thu y Qudc gia.
- Gilng vi khuan: Cac chung vi khuln A.
pleuropneumoniae, P. multocidd va S. suis chuan va cac huyet thanh chuan tuong ung do Nhat Ban cung cap.
- Cac loai thuoc dieu tri viem phdi do vi khuan nhu: Khang sinh the he mdi; thuoc bo, thudc trp sue, trp lye; thudc ha sot, khang viem,...
* May moc thiet hi
Budng cay vd triing, tii am, tu lanh, tii say, noi hip, may ly tam, may lie gian, may nhan gen PCR, ELISA va mot sd dung cu, thiet bi khac dung cho nghien ctru.
Npi dung nghien cihi
- Xac djnh Serotype eiia cac chung vi khuSn A. pleuropneumoniae phan lip.
- Xic dinh dpc lye ciia cac chung vi khuln P.
multocida phan lap.
- Xac dinh tinh man cam khang sinh ciia cdc chung vi khuan S. suis phSn lap.
Phuong phap nghien cuu
* Phuffng phdp xac dinh Serotype cua cac chung vi khudn phdn lap
Xac dinh Serotype cua vi khuan A.
pleuropneumoniae bang phan ling ket tiia khulch tan tren thach (AGID); P. multocida va S. suis dugc thuc hien bang cac phan ung ngung ket va phan iing PCR da dupe chuin hda cua Bd mdn Vi triing, Vien Thiiy Qulcgia[6].
* Phuong phap xdc dinh so lupng vi khudn Mau canh khuan cSn xac djnh dupe lay v5 triing, sau do pha loang canh khuln vdi niroc muli sinh ly vd triing thinh cac dp pha loang khic nhau hi 10"' din 10"*. Dung 3 dp pha loang 10"*, 10"^ 10'* moi dp pha loang cay tren 3 dia thach TSA (1-3% YE), mli dia nho 0,1 ml, dan diu tren mit thach. Sau 18 gia nuoi cly d tu Im 37°C cd 5% CO2, tiln hanh dem so lupng khuan lac mpc d cac dia thach.
Sd lupng vi khuan trong 1ml canh khuSn dupe tinh theo cdng thuc dudi day vdi don vj tinh li CPU:
X = a x N x l O Trong do:
X: So lupng te bao vi khuan trung binh tronj 1ml canh khuan.
N: He sd pha loang.
a; Sd khuan lac trung binh cd trong 0,lffl canh khuan pha loang.
* Phuong phap xac dinh ddc lye cua ca chung vi khuan phan l?.p
Xac dinh dpc luc ciia cac chiing / pleuropneumoniae, P. multocida vi S. su.
phan lap duac tren d^ng vat thi nghi?i
Nguyin Thj Kim Lan vd Dtg
Tap chi KHOA HQC & C 6 N G N G H ?164(04): 87-95 (chudt nhat tring) theo phuang phap thudng
qui ciia Bd mdn Vi triing, Vien Thu y QuIc gia [6].
* Phuang phap xac dinh min cam khang sinh eua cic chdng vi khuln phan l|p Do dudng kinh vdng vd khuan va so sanh vdi bang chuan dl danh gia miic dp min cim khang sinh cua cac chung vi khuln phan lap.
Phuvng phap xir ly s l lieu
Cic sd lieu thu thap dupe xii ly theo phuong phap toan hpc thdng dyng va thing ke. sinh hpc cua Nguyin Vin Thien (2008) [7], tren phan mlm Excel 2007 va Minitab 14.0.
KET QUA NGHIEN CUtJ
Ket qua xac djnh Serotype cua mpt s6
chiing vi khuan A. pleuropneumoniae, P.multocida va S. suis phan lap dup'c
Da tien hanh xac dinh Serotype huyet thanh hpc cua 11 chung vi khuin A.
pleuropneumoniae, 16 chiing P. multocida va 17 chung 5. .sHw phin Tip duac bang phin ling kit tiia khuech tan tren thach (AGID) vdi khang huyet thanh chuin do Nhat Ban chi tao. Ket qua dupe trinh biy d bing 1.
Bang 1 cho thiy; Trong II chiing vi khuan A. pleuropneumoniae phan lap cd 7/11 chung thudc serotype 2, chiem ti le 63,64%.; 4/11 chung thupc serotype 5, chiem ti 1? 36,36%.
Trong 16 chung vi khuin P. multocida phan lap dupe cd 8 chung thudc serotype A, chiem 50% va 8 chung thupc serotype D, ehiem 50%. Trong 17 chung vi khuan S. suis phan l§p dupe, sd chiing thudc serotype 2 li 7/17 chung, chilm ti le cao nhit (41,18%); 10 chiing con lai thudc serotype 7 va 9 vdi ti Ip tuong duong (29,41%).
Bdng ddnh gid miic do mdn cdm cita vi khudn vai mot s6 logi khdng sinh (NCCLS - 2002)
TT 1 2 3 4 5 6 7 8 9 10 II
Loai khang sinh Ceftiofur Ampicillin Amoxicillin Neomycin Amikacin Gentamicin Lincomycin Colistin TeUacyclin Erythromycin Florfenicol
Ham lupng 30 ng 10 (ig 20 (ig 30|Jg 30 tig 10 pg 15 ng 10 (ig 30 (ig 15 ng 30 ng Bang 1, Kit qud xdc dinh Serotype
Du'&ng kinh vdng vo khu Manh
> 2 1
> 2 2
> 2 0
> 1 7
> 1 7
> 1 9
> 1 5
> 1 5
> 2 9
> 2 1
> 2 3
Trung binh
cua mQt so chiing vi khudn P multocida va S suis phdn A. pleuropneumoniae
Serntvnp n = 11 So chung (+) Serotype 2 7 Serotype 5 4 Serotype 7
Serotype 9 Serotype A Serotype D
chung TII4(%)
63,64 36,36
lap duoc P. multocida n = 16 chung So chung (•^)
%
8 TilfC/.)50,00 50,00
1 8 - 2 0 1 9 - 2 1 1 3 - 1 6 1 5 - 1 6 1 3 - 1 4 1 3 - 1 4 2 6 - 2 8 1 6 - 2 0
I n (mm) Khang thuoc
< 1 7
< 1 8
< 1 9
< 1 2
< 1 4
< 1 5
< 1 2
< 1 2
< 2 5
< 1 5
< 2 0 A. pleuropneumoniae.
n S. suis
= 17 chiing So chung (+} rilf(%)
7 5 5
41,18 29,41 29,41
-
Nguyen Thj Kim Lan vd Dtg p cm K.HUA nv»^ Oi. t^wii Ket q u i nghien cuu cua chung tdi tucmg ddng
vdi k i t qua nghien cuu cua mpt sd tac gia khac. Cu Him Phd va cs. (2005) [5] cho bilt, trong cac Serotype cua vi khuan A.
pleuropneumoniae p h i n lap dupe tir lpn mac benh viem dudng hd hap tai mpt so dia phuong m i i n B i c Viet N a m thi Serotype 2 luon e h i l m uu t h i . Kang-Hee O va cs. (1990) [9] phan l i p vi khuan tir 155 m i u phoi cua lpn thit, ket q u i cho thay, trong so chiing vi khuan P. multocida phan lap duoc c6 60,4%) sd chiing thupc Serotype A; 18,6%) sd chung thupc serotype D va 21,0%i so chung chua xac dinh d u g c serotype. Lun Z. R. va cs, (2007) [10] cho bilt: Serotype 2 cua cac chung vi k h u i n S. suis d u p e xac dinh l i Serotype thudng gap nhat gay b S n h c h o lpn va ngudi d hau bet cac n u d e tren the gidi.
K e t q u a kiem t r a dpc l u c ciia cac c h u n g vl khuan A. pleuropneumoniae phan lap dupe Xac dinh dpc luc cua 45 chung vi khuan A.
pleuropneumoniae phan lap t u phdi va cudng hpng cua lpn tai tinh Bac Ninh, mdi chung vi khuan A. pleuropneumoniae d u p e tiem vao phuc xoang cho 2 ehupt nhat trang, su dyng canh trung da nudi cay 24 gid d 37°C trong d i l u kien cd 5 - 10% C O j , l i l u tiem 0,2 ml/con; theo ddi chupt trong vdng 36 gid sau khi tiem. Ket qua kiem tra dpc lye dupe trinh bay d bang 2.
Ket qua d bang 2 cho thay: Trong 90 chupt dupe tiem canh khuan A. pleuropneumoniae vao phiic xoang co 87 chu6t chet trong vong
Bang 2. Kit qud kiim tra doc luc cua cdc
12 - 36 gid; chi cd 3 chudt khdng chet. T ^ c4 so chupt tiem canh khuan cua cac chung A.
pleuropneumoniae phan lap tir lpn d BIc Ninh d i u chet trong vdng 36 gid cdng cudng dSc Rieng cac chung phan lap tir lpn nuoi tai TP.
B i c Nmh v i huySn Gia Binh cd ddc luc cao, gilt c h i t 100%) sd chupt y o n g vong 24 gia.
Cac chung A. pleuropneumoniae thu thap tai huyen Q u i Vo, Tien D u va Yen Phong co doe lyc t h i p ban, lam chet 90 - 93,75%i sd chuOt.
K i t qua nghien c u u cua chiing t6i tuong dong vdi vdi k i t qua nghien cuu Le Van Dosmg (2013) [2] da k i i m tra dpc lye ciia 12 chung vi khuan A. pleuropneumoniae thu thap d lpn nuoi tai tinh S l c Giang, t h i y 9 chung c6 dpc luc manh, g i l t chet 100% sd chupt thi nghian trong k h o i n g thdi gian tu 12 - 36 gid; 3 chung gilt c h i t 50%> sd chupt thi nghiem trong k h o i n g thdi gian tir 18 - 48 gid.
N h u vay, vi khuan A. pleuropneumoniae phan lap CO dpc luc cao Vdi dpc luc cao nhu vay thi cac chung vi khuan nay la nguyen nhan quan trpng g i y viem phoi cho dan lpn ciia tinh Bac Ninh.
Xdc dinh doc Ittc cua cdc chung vi khuan P.
multocida phan lap
TiSn hanh xac dinh dpc lyc cua 67 chung P.
multocida tren chupt nhat trang. Moi chung tiSm cho 2 chudt vao phuc xoang, vdi lieu 0,2 ml canh tning nudi c i y vi khuan d 37°C/24 gid; sd chupt thi nghiem d u a c theo doi hong vdng 36 gid. Ket q u i d u p e trinh bay d bang 3.
chung vi khudn A. ephdn lap Dia phirang
TP. Bae Ninh QueVo Thuan Thanh Luong Tai Tien Du Gia Bmh Yen Phong Tir Son
Tinh chung s l c h u n g kiem tra (n)
4 8 6 7 6 5 5 4 45
So chuot
tiem (con) 8 16 12 14 12 10 10 8 90
Lieu ti@m phuc
\oang (ml/con)
0,2 0,2 0,2 0,2 0,2 0,2 0,2 0,2 0,2
< 1 2 h 0 0 0 0 0 0 0 0 0
sd chu$t ch^t 1 2 - I8h
4 6 6 8 2 8 2 0 36
: sau tiSm (con)
> 1 8 - 2 4 h 4 2 4 4 8 2 4 6 34
> 24 - 36h 0 7 2 2 1 0 3 2 17
Til? chuot
chet
(%) .
100 93,75
100 100 91,67 100 90,00 100 96,67_
Nguyen Thj Kim Lan va Dtg Tap chi KHOA HOC & CONG NGHE 164(04): 87-95
Bang 3. Kit qud kiim tra dpc luc ciia cdc chung vi khudn P. multocida phdn lapDja phuorng
, , So chupt Lieu tiem _ chuns; ,.. , ,
, .1 ° tiem phuc xoang tra°(") '°°°' ' " ' " " ° '
So chuot chit sau tiSm (con) : 12h 12 - 18h
Til?
chupt lS-24h >24-36h '>•'*
(%)
TP. Bac Ninh QueVo Thugn Thanh Luang Tai TienDu Gia Binh Yen Phong Tit San
Tinh chung 5 7 5 10 12 14 9 5 67
10 14 10 20 24 28 18 10 137
0,2 0,2 0,2 0,2 0,2 0,2 0,2 0,2 0,2
0 0 0 0 0 0 0 0
0
49 53
2 3 6 3 6 4 0 4 28
90,00 92,86 100 90,00 100 100 100 100 97,01
Qua bang 3 cho thay: Cae chung P. multocida
phan lap tir Ipti d huyen Thuan Thanh, Tien Du, Gia Binh, Yen Phong va Tii Son cd doe lyc cao, gay chet 100% sd chupt thi nghiem trong vdng 1 2 - 3 6 gid sau khi cdng cudng dpc. Cac chiing thu thip td TP. Bic Ninh, huyen Que Vd va huypn Luang Tai giy chet 90 - 92,86%. s6 chudt trong vdng 36 gid. Ket qui cua chung toi tuong ddng vdi nghien ciiu cua Do QuIc Tuan va Nguyen Quang Tuyen (2007) [8]. Khi kiim tra dpc lyc cua cic chiing vi khuan P. multocida phan lap dupe d lpn tai tinh Thai Nguyen, Tuyen Quang va BIC Kan, thay cac chdng P. multocida deu co dpc lyc msinh, gay chit chupt thi nghiem tir 50 - 100% trong thdi gian tir 20 - 48 gid.
Nhu vay, cac chung vi khuan P. multocida mi chiing tdi phan lap tir lpn d Bac Ninh deu cd dpc luc cao, vdi dpc luc niy chiing hoan toin cd kha nang giy viem phdi cho lpn khi gap dilu kien thuin Ipi, sue dl khing cua lpn giam do thdi tilt thay dli, cham sde nudi dudng kem hoac bi mac PRRS.
Xdc dinh dpc ly:c cua cdc chung vi khuan S.
suis phan lap
Kiim tra dpc lyc cua 88 chung vi khuan S.
suis tren chudt nhit tring, moi chupt dupe tiem 0,2 ml canh trung nudi cay d 37°C/24 gid vao phiic xoang. Chupt thi nghi?m duoc
theo ddi trong vdng 36 gid; khi chudt chet duac md khim, kiSm tra benh tich va phan lap lai vi khuan tir mau tim. Ket qui dupe trinh bay d bang 4.
Qua bang 4 cho thay: Trong 176 chudt dupe tiem canh khuln cd 168 chudt chit trong vong 36 gid sau cdng cudng dpc, chiem ti le 95,45%>. Cac chiing vi khuan 5*. suis phin lap tir lpn tai huyen Que Vd, Luong Tii, Gia Binh va Tir San ed dpc luc cao, giy chet 100%) so chudt thi nghiem trong vdng 12 - 36 gid. Cac chung phan lap tir Ipn d TP. Bac Ninh, huyen Thuan Thinh, TiSn Du va Yen Phong CO dpc lyc thap hon, gay chet 87,59 - 90%) sl chudt trong vdng 36 gid sau cdng cudng dpc.
Nhu vay, cac chung S. suis phan l|.p dupe tir lpn tai tinh Bic Ninh deu la nhiing chung cd dpc luc khi cao, gay chet chupt thi nghiem trong thdi gian ngan.
Ket qua theo doi khang sinh do ciia cac
chung vi khuan A. pleuropneumoniaeChung tdi da tien hanh kiem tra mire dp man cam vdi m0t sd khang sinh eua cac chung A.
pleuropneumoniae phin lip dupe, tir do lya chpn loai khang sinh man cam manh de xay dyng phac do dieu tri bpnh viem phoi cho Ipn hieu qua. Kit qua dupe trinh bay d bing 5.
91
Nguyen Thj Kim Lan vd Dtg 1 ap cm RJiUA HV>.^ oi. i^L/iivj iNvjnij lOinutj: HI-%
Bang 4. Ket qud kiim tra dgc l\fc cita cdc chiing vi khuan S. suis phdn Igp
Dja phirong
TP B5c Ninh Que Vo Thu|n Thanh Luang Tai TiSn Du Gia Binh Yen Phong Tir Son
Tinh chung
Si
chiingkiem tra (n) 9 10 12 11 16 10 10 10 88
si
chuQt tiem (con) 18 20 24 22 32 20 20 20 176
Lieu tiem phiic xoang (ml/con)
0,2 0,2 0,2 0,2 0,2 0,2 0,2 0,2 0,2
So chu$t ch
O ^ " " l 8 h 0 8 0 4 0 12 0 4 0 12 0 6 0 8 0 10 0 64
et sau tiem (c
> 1 8 - 24h 7 8 5 10 6 6 8 8 58
on)
> 2 4 - 36h 2 8 5 8 10 8 3 2 46
Till ehu$t chet
(%)
94,44 100 91,67 100 87,50 100 95,00 100 95,45 Bing 5. Kit qud theo doi khdng sinh do ciia cdc chiing A. pleuropneuinomae phdn lap
Loai khdng sinh , So chung
'T^^ (n)
(^g) ^ '
Muc do n
Trung binh Khdng thuoc
CeftiofiirAmoxicilin Florfenicol Ampicillin Tetracyclin Colistin Gentamycin Neomycin Erythromycin Lincomycin
30|ig 20 ng 30 ug 10 ng 30 (Ig lOllg 10 ng 30 ng 15 ng 15 ng
45 45 45 45 45 45 45 45 45 45
34 32 31 25 19 5 3 0 0 0
75,56 71,11 68,89 55,56 42,22 11,11 6,67 0 0 0
6 7 5 9 14 21 22 10 8 3
13,33 15,56 11,11 20,00 31,11 46,67 48,89 22,23 17,78 6,67
5 6 9 11 12 19 20 35 37 42
11,11 13,33 20,00 24,44 26,67 42,22 44,44 77,77 82,22 93,33 •
Qua bing 5 cho thay: cac chung A.
pleuropneumoniae phan lap dupe man cam manh nhat vdi Ceftiofiir, chiem ti le 75,56%.;
tiep theo la amoxicillin, Florfenicol va Ampicillin vdi ti le man cim manh lan lupt li 71,11%; 68,89% va 53,33%. Mdt s6 khang sinh dang dupe sd dyng phd bien cd ti le khang thudc kha cao nhu lincomycin bi khang tdi 93,33%i; tiep den la erythromycin va neomycin vdi ti le chung khang thudc tuong ung la 82,22%> va 77,78%). Co nhieu nguyen nhan dan den hicn tupng khang khang sinh cua VI khuan A. pleuropneumoniae ngay cing gia tang nhu viec diing khang sinh dieu tri keo dai, khang sinh thudng xuyen dupe bd sung vao thiic in chan nuoi, do hien tupng di truyen tinh khang thulc bdi cae gen nim
trong plasmid ciia vi khuan A, pleuropneumoniae.
Ket qua nghien ciiu ve khi nang man cam khang sinh eua cac chdng vi khuan A.
pleuropneumoniae phan llip d lpn cua tinh Bac Ninh phii hop vdi ket qua nghien cuu ciia Nguyen Thj Thu Hing (2010) [4].
Ket qua theo doi khang sinh do cua cac chiing vi khuan P. multocida
Zarfel G. va cs. (2014) [12] cho bilt, trong nhiing nam gin day, vi khuan khang da thuoc da dupe phat hien ngiy cang nhilu. Vi v|y, viec xac dinh kha nang min cam ciia cac chung vi khuan P. multocida vdi mpt so khang sinh la rat can thiet. Kit qua dupe trinh bay d bang 6.
92
Nguyin Thi Kim Lan va Dtg Tap chi KHOA HQC & CONG NGH$ 164(04): 8 7 - 9 5 Bang 6. Kit qud theo doi khdng sinh do cua cdc chung vi khudn P. multocida phdn lap
j j a m ^ _ Muc dp miin cam Lo?i khang sinh lupng
(^tg)
So chiing
(n) Trung binh Khdng thuoc
Cettiofiir Florfenicol Amoxieilin Ampicillin TeUacyclin Gentamycin Colistin Neomycin Erythromycin Lincomycin
30 ng 30 ng 20 ng 10 ng 30 ng 10 ng 10 ng
30 ng
15 pg 15 ng67 67 67 67 67 67 67 67 67 67
58 56 49 41 27 13 9 5 0 0
86,57 83,58 73,13 61,19 40,3 19,4 13,43 7,46
0 0
7 5 11 12 20 26 28 15 19 12
10,45 7,46 16,42 17.91 29,85 38,81 41,79 22,39 28,36 17,91
2 6 7 14 20 28 30 47 48 55
2,98 8,96 10,45 20,90 29,85 41,79 44,78 70,15 71,64 82,09
Bang 6 cho thiy: Cac chiing vi khuan P.
multocida phan lap dupe min cam manh vdi Ceftiofiu (86,57%), Florfenicol (83,58%), amoxicillin (73,13%.) vi AmpieiUin (61,19%i). Rat nhieu chung da khang lai mdt sd khang sinh nhu lincomycin (82,09%.), erythromycin (71,64%) va neomycin (70,15%). Truong Quang Hii vi cs. (2012) [3] da xac dinh khi nang man cim vdi khing sinh cua cic chung vi khuin P. multocida phan lap dupe d Ipn mac benh viem phdi va cho biet, vi khuan P. multocida man cam manh vdi Ceftiofiir (95,0%.), Florfenicol (90,0%), Amoxicillin (80,0%.), Ofloxacin (75,0%), vi Streptomycin (70,0%); khang lai
Bang 7. Kit qud theo doi khdng smh do cua i
vdi mpt sd loai khang sinh nhu Neomycin (70,0%), Penicillin G (65,0%) vi Tetracyclin (60,0%).
Nhu vay, cac chung vi khuin P. multocida phan lap dupe d lpn.tai tinh Bac Ninh man eim manh vdi Ceftiofiir-,va Florfenicol; da khing lai mpt sd kh^ng sinh nhu Lincomycin, Erythromycin va, Neomyci|i. Ket qui nay tuong ddi phii hpp vdi ket qua nghien cihi cua tic gii tren.
Ket qua theo doi khang sinh do ciia cac chung vi khuan Streptococcus suis Ket qui dupe trinh bay d bang 7.
dc chung vi khudn S. suis phdn Igp dupe , . . . . Ham
Loai khang , • . . lupng smh , ^
Muc dp man c§m So chung
(n) Trung binh Khang thuoc
Ceftiofiir Amoxicillin Florfenicol Ampicillin Tetracyclin Gentamycin Lincomycin Neomycin Erythromycin Colistin
30 ng 20 Mg 30 ng 10 pg 30 pg 10 pg 15 pg 30 pg 15 ng 10 pg
72 70 67 62 41 17 8 0 0 0
81,82 79,55 76,14 70,45 46,59 19,32 9,09
0 0 0
9 7 13 9 11 33 36 28 23 5
10,23 7,95 14,77 10,23 12,50 37,50 40,91 31,81 26,14 5,68
7 11 8 17 36 38 44 60 65 83
7,95 12,50 9,09 19,32 40,91 41,18 50,00 68,19 73,86 94,32
rsguyen ini is.im i-aii vu i:/ig lojj i-m !.•,-».w Qua bang 7 cho t h i y : cac chung S. suis p h i n lap t u lpn d tinh B i c Ninh m i n c i m manh vdi Ceftiofiir (81,82%), Amoxicillin (79,55%), Florfenicol (75,00%) v i Ampicillin (70,45%);
cac chiing nay da Idiang lai mot sd khang sinh nhu Colistin (94,32%), Erythromycin (73,86%) va Neomycin (68,18%). K i t qua cua chung toi cung tuong ddng vdi ket q u i nghien cuu cua Tieu Quang An v i N g u y i n Huu N a m (2011) [1]: trong 11 chiing vi k h u l n S. suis phan lap dupe d lpn d u a n g tinh vdi PRRSV cd 63,64% sd chung m i n c i m vdi mpt s l kliang sinh nhu Amoxicillin, Ampicillin, Florfenicol va da khang manh vdi mdt sd khang sinh nhu Enrofloxacin va Colistin.
Nhu vay, cac chung vi khuan S. suis phan lap dupe d lpn tai tinh Bac Ninh man cam manh vdi Ceftiofiir, Amoxicillin, Florfenicol va khang lai mpt sd khang sinh n h u Colistin, Erythromycin, Neomycin v i Lincomycin.
K E T LUAN
C i c chung vi k h u i n A. pleuropneumoniae p h i n lap dupe thupc Serotype 2 (63,63%) va Serotype 5 (36,36%). Cac chung vi khuan P.
multocida p h i n lap dupe thupc Serotype A (50,00%) v i Serotype D (50,00%). Cae chung vi khuan S. suts phan l i p dupe thupc Serotype 2 (41,18%), Serotype 7 (29,41%) va Serotype 9 ( 2 9 , 4 1 % ) .
Cac chung vi khuan p h i n lap d u p e deu cd dpc lyc cao, gay chet 95 - 100% sd chudt thi nghiem sau khi cdng cudng dpc 12 -36 gid.
Cac chung vi khuan A. pleuropneumoniae, P.
multocida v i S. suis p h i n lap dupe de man c i m m^nh vdi Ceftiofur, Amoxicillin, Florfenicol.
T A I L I E U T H A M K H A O 1. Tieu Quang An, Nguyen Huu Nam (2011),
"Xac dinh mpt so vi khuan ke phat gay chet lpn trong vCing djch lpn tai xanh d huyen Van Lam linh Hung Yen nam 2010", Tap chi Khoa hoc ky thuat thuy. tap 18(3),tr. 5 6 - 6 4 .
2. Le Van Duong (2013), Nghien ctru mot so dac tinh sinh hgc cua vi khudn Actinobacillus pleuropneumoniae, Pasteurella multocida.
Streptococcus suis gay viem phoi trong Hgi chOiig rdi loan ho hap vd sinh sdn a Ign tai Bac Giang biin phdp phong tri, Lu?in an tiln sT Thii y, Tniong Dai hpc Nong Lam - Dai hpc Thai Nguyen.
3. Truong Quang Hii, Nguyin Quang Tmhj Nguyen Quang Tuyen, Cu Hiiu Phii, lA Vin Duong (2012), "Ket qua phan l^p va xac djnh in$t sl dac tinh smh hoc ciia cac chung Streptococae suis va Pasteurella multocida d Ipn mlc viSm phfii t^i tinh Bac Giang", Tap chi Khoa hpc ky thuqt Iku y, tap 19(7), tr. 7 1 - 7 6 .
4. Nguyin Thi Thu Hing (2010), Nghien cuu m&
so dac tinh sinh hpc vd tinh sinh mien djch cua Actinobacillus pleuropneumoniae phdn Iqp tit ign ldm ca sd cho viec che tao vaccine, Luan an tien si Nong nghi?p, Vi^n Thii y Quoc gia. Ha N$i, t.
115-116.
5. CuH&uPhu, Nguyin Ngpc Nhien, Nguyin Thu Hing, Au Xuan Tuin, Nguyin Bich ThuJ, Vfi Ngpc Qu^, Ph^m Bao Ngpc (2005), "Xac dinh nguyen nhan gay b?nh dudng ho hap ciia lpn nuoi tai mot so tinh phia Bac", Tqp chi Khoa hoc ky thuat Thuy. tap 12(4), U. 23 - 32.
6. Cii Huu Phd (2011), Nghien ciru mdi liin qimit giira Hpi chimg rdi loan hd hap vd sinh sdn d lpn vdri vi khudn gay b^nh ke phdt vd xdc djnh bi§n phdp phong, trj binh, Bao cao khoa hpc, Vi§n Thii y Quoc gia, 2011
7. Nguyin Van ThiSn (2008), Phuang phap nghiin cihi trong chan nudi, Nxb N6ng nghiep, Ha Noi.
8 Do QuIc TuIn, Nguyin Quang Tuyen (2007),
"Ket qua kiim tra doe lyc va tinh man cam khang sinh cua Pasteurella multocida phSn Igp dupe tit lpn tai khu vuc mien niii phia bic". Tap chi Khoa hpc ky thudt Thu y, tap 14(6), tr. 46 - 51.
9. Kang-Hee O., No-Chan Park, Lee-Zun Kim, Duk-Sang Park (1990), "Serogruop and dnig susceptibility of Pasteurella multocida pneumonia in pig", Kprean J. Vet. Serv.. 13(1), pp. 69-74.
10. Lun Z. R., Wang Q. P., Chen X. G., Li A. X Zhu X. Q. (2007), "Streptococcus suis: w emerging zoonotic pathogen". Lancet Infect Dis 7(3), pp. 201-209.
11. Reed M. J., Muench H. (1938). "A simpl method for estimating fifty percent end points"
Am J Hyg. 27, pp. 493-497.
12. Zarfel G., Galler H., Luxner J., Pettemf C , Reinthaler F. F., Haas D., Kittinger C , Grisol A. J.PIess P., Feierl G. (2014), "Multiresistai bacteria isolated from chicken meat in Austria' Int. J Environ. Res. Public Health, 11(12), pi 12582-12593,
94
Nguyen Thi Kim Lan vd Dtg Tap chi KHOA HQC & CONG NGHB 164(04): 87 - 95
S U M M A R Y
I D E N T I F Y S E R O T Y P E , V I R U L E N C E A N D A N T I B I O T I C R E S I S T A N C E O F B A C T E R I A L S T R A I N C A U S I N G P N E U M O N I A I N P I G S I N B A C N I N H P R O V I N C E
Nguyen Thi Kim Lan", Nguyen Thi Ngan, Nguyen Van Quang, Phan Thi Hong Phuc, Le Minh, Pham Dieu Thuy, Tran Nhat Thang, Duong Thi Hong Duyen University of Agriculture and Forestry - TNU Results identified Serotypes, vimlence and antibiotic susceptibility of three bacterial strains that caused pneumonia in pigs in Bac Ninh province, shown that: The isolated bacterial strains of Actinobacillus pleuropneumoniae (A. pleuropneumoniae) were Serotype 2 (63, 63%) and Serotype 5 (36.36%); the bacterial strains of Pasteurella multocida (P. multocida) were Serotype A (50.00%) and Serotype D (50.00%); Streptococcus suis strains {S. suis) were Serotype 2 (41.18%), Serotype 7 (29.41%) and Serotype 9 (29.41%). The isolated bacterial strains are highly vimlent and have the lethality from 95 tolOO% of experimental mice after poisoining the virulent 12-36 hours. The isolated strains of A. pleuropneumoniae, P. multocida and S. suis were sUongly susceptible to Ceftiofur, Amoxicillin and Florfenicol antibiotic medicine.
Keywords: Serotype, virulence, antibiotic resistance, bacteria, pigs.
Ngdy nhan bdi: 18/01/2017; Ngdyphdn bi^n: 10/02/2017. Ngay duy?t ddng; 27/4/2017
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