TTLFNHI £MVA

HOI NGHI KHOA HQC CONG NGHE SINH HQC TOAN QUOC 2013 ~

" TTLFNHI £MVA;SI?T5 rKH^NG"KHANG~STNH"COA^rKHUAN 'VtBRIO~~

CHOLERAE PHAN LAP TU" HUY£T HEO, NGHEU VA PHAN BENH NHAN T | £ U CHAY TAI TINH TRA VINH

Nguyen Thi Oku\ NguySn Thfty Linh\ Hb Thj Wet Thu^ ' ' Trudng B9I hgc Trk Vinh

'Trudng D9I hgc Cin Tha

.T6M TAT

KJt qua da phSn ISp vi khula Vibrio choleae tren 300 man, bao gom 160 mlu nghSu; 100 mlu huy^ heo t^i eic co sd giit mfi vi 40 miu phSn trSn b^nh tMa tiSu chiy tgi blah v i ^ da khoa tinh Tra Vinh, ti 1? phan l$p V choleae \ka lu^t trSn nghSu U 10%, trSn huy^ heo co pha nude tgi co so gilt mk la 4%, chua cfi d ^ hi^u duong tinh tren b§nh nhSn tiSu chay tgi tinh Tra Vinh.

K^t qua khing sinh dS cho thiy da s6 vi Uiuln V. choleae nhgy v6i norfloxacin (100%), v*i chloramphenicol (70%), vi dh khang v6i hoin toin v6i amoxicillin (90%). Cf 15% mau duoi^ tinh v6i khang huySt thaih da gii (0139, Ogawa, Inaba), Il \f duong tinh voi khang huy^t thanh don gji Ogawa (10%) vi vdi khang huyet thanh don gii Inaba (10%) tren nhung mlu b ^ pbiin dugc phin lip.

Tir khoa: Vibrio choleae; nghSu; ti 1§ nhiim; Sk khing khing sinh; Tri Vmh MdoAu

Th^ giai da tr^l qua 7 d^i djch t i , tCrnam 1817 d^n nam tOlZdk c6 6vydaidjchxSyra, nhOngd^idjchndyd^ubStdau tir An DO vk dl6u do VibtUi cholerae 01 type slnh h(?c c6 ffiln gSy ra. D?i dfch lhi> 7 khke vd\ 6 flgi dfch tru^c. d^i djch nky do V.cholerae type sinh hpc El Tor gSy ra vk c i ngu6n g6c tO- dao Celebes cOa Indonesia nSm 1961. Dgi djch nky kfio dki nhlt v&'cfl phgm vl r^ng hem 6 dgi d|di tru'd'c dd. din nay c6n nhifiu nu&c thfing bko nhO'ng dgt bdng phit djch tk cDng do nguyfin nhSn nky gky ta (V5 V3n Luvng, 2009).

Rieng Vi$t Nam da xay ra 3 dgt djch lieu chiy d p nguy hi^m: dgt thCt nhk tir ngky 23/10 - 6/12/2007 a 14 tinh, thanh phia Bac vdi 1.878 ca, trong dk ed 295 htr&ng hgp duang tinh vcri vi khuSn ta; dgt thi> hai h> ngiy 24/12/2007 - 5/2/2008 6 Hk NOi v6i 58 ca, ciS 32 ca do vi khuln t i ; dgt thO ba, tir ngky 6/3 den 11/4/2008, dk c6 1.335 ca, trong do 136 ca duo-ng tinh vdi vl khuln t i , d 18 flnh, thinh thu^c c i 3 mien Bic, Trung, Nam (V8 Vin Lu'gng, 2009) Nam 2009-2010, djditi Igl xu^thi$ntai cac tinh mifin BacVi#INam, m$tchungvi khuln V. c/io/s/a 0139 dirge phin l^ptO 7 mau ninic v i du^c dgt tfin l i V. cholerae 0139 (Dong Tu Nguyen et al, 2012).

Tinh Trk Vinh c6 vj tri dja ly vd\ nhlfiu nguy ca fa'fim In b#nh djch la vi cd b£r bifin keo dii khoing 65 km v i trfin dja bin Tri Vinh' od h$ th6ng sflng chinh vdi tfing chlfiu dii 578 km, tfong dfi cfl cic sflng l*n l i s6ng H^u, sflng C6 Chifin v i sflng Ming Thit, vl thfi rlt d l cho '^^c lu-u hinh vi khuln ta ti> sflng Cfi Chi&n,biln Duyfin Hal v i Clu Ngang.

Dfi ngin ch$n sg lay lan v i nguy hifim cCa b$nh djch ta, d$c bi$t la a viing dfing bing sfing Ciru Long cd nhifiu difiu kifn dfi vi khuln liy lan, chOng tfli nghifin cihj "TT lfi nhifim v i s^ dfi khang khing sinh cOa vi khuln V.cholerae trgn huyet heo, nghfiu y i phin bfnh nhin lifiu chkyt^l finh Tri Vinh", vdi myc lifiu xic djnh IT 1$ nhifim V. cholerae trSn cSc logi mlu phin l$p| xic djnh serotype^ phfi bifin it/u hanh cfl Ihl gay bfnh cho ngu'fl'i v i tinh nhgy cam khing sinh cda cic chung V. cholerae phkn l|p du'gc.

VAT L1|U VA PHl/CTNG PHAP Vft lieu

Cic I09I mfli trifdng bio quin mlu v i nufll cly vi khuan Vibrio chderae: Cary - Blair, Alkaline saline peptone water (ASF=W), Thiosulfate-Citrat-Bllei-SacchanDse (TCBS), Saline Nutrient Agar (SNA).

Mlu phin bfnh nhin tieu chiy: du-gc thu lh@p sdm, bao quan cr moi t n j ^ g vSn chuyfin Caiy - Blair, v i chuyen vfi

phflng thi nghifm dfi nufli cly. phin l|p. ^ Mlu nghfiu: n>a sgch chit bfin bfin ngoii, lly phfin thjt d l nhuyen (1 g) ling sinh Irong 9 ml mfli Irudng nude pepton

mufii kilm ASPW.

Mau huylt heo; khoing 5ml/mlu dugc lly Igi cac co sd gifit mo, cfl bfi* sung nu-dc mufii 2%, mlu dugc chuyfin vfi phflng thi nghifm ting sinh tnsng 9 ml mfli Irudng nude pepton mufii kifim APW.

Tit c i cic logl mlu dfiu dugc nudi cly tich IQy (enrichment) trong moi trudng APW tie 8 -12 gid.

Phuorng phip

Xic dinh V. cholerae bing phin dvg slnh hda

Mlu dugc nufli d y tfch lOy trong mfli trudng Peptone kifim (APW), sau dfl d y chuyen Irfin mfli trufl-ng chuyfin bifit TCBS, khuln Igc cfl thfi cfl hal logi, miu ving ho|c xanh tuy kiii.

Chgn khulh Igc V. cholerae (kfch thufl'c 2 - 3 mm, "cd miu ving, xung quanh cfl quing trlng dye, bdng) d y sang mfii

Irudng SNA nhlm lim ting s l khuln Igc cOa vi khuln. Trfin SNA khuln Igc Vibrio spp. trdn, Iron, bdng, miu trlng sO-a.

H O I N G H I K H O A HQC C O N G N G H E S I N H H Q C T O A N Q U 6 C 2 0 1 3

K h u l n Igc trSn dirge chpn de thi> nghi?m oxidase duong tinh (+); TSI d u o n g tinh (+), khong sinh hoi v i khflng sinh HaS;

Indole duong Unh {•^).

T h u tfnh u a m j n cua V. chderae d cac nfing dO muoi NaCI k h i c nhau (0%, 2%, 6%, 8% va 10%), Vibrio cholerae mpc tfil t r i n mfli trudng cd 2% m u l i .

Phuong phip djnh danh bang phan dng huyet thanh hgc.

K h i n g h u y l t thanh c h i n d o i n vi k h u l n V. chderae bao gfim:Khing huyel thanh t i da g i i 3 chCing: Inaba. Ogawa v i 0139; k h i n g huyfil thanh l i don g i i Inaba; k h i n g huyfit thanh l i don gia Ogawa.

Bang 1. Tdm t i t lhi> nghifm khing huyet thanh V. cholerae

Khdng nguyfin Khing huy^ Khing huylt Dung iQch muAi Ket iu^n thanh kiaba Ihanh Ogawa sinhl^

V. chderae Ot * - - Chung Inaba V. cholerae Ot - + - ChOng Ogawa V. cholerae 0 1 t + - ChOng da g i i V.eholarae Ot - _ - - Chung khdng dgc hi$u

X i c djnh k h i n g slnh d i v i nfing d9 i>c che t i l Ihieu (MIC: Minimum Inhibitory Concentration) cua k h i n g sinh d f i i vd^ Vibrio cholerae. • ' • Phtrcmg phip khing slnh do (Kliiiy-Bauer)

Chgn 05 logi k h i n g slnh Ihlch hgp dfli vdi vi k h u l n duflng rudt thudng d u g c s u dyng dfi difiu tri b f n h dich t i . Iheo tifiu c h u i n cda CLSI (Clinical and Laboratory Institute, 2010).

Phuong phip tim gli trf ndng tfj i>c ch4 t&l thieu cua khing slnh (MIC)

T h i xfit nghifm l i m k h i n g sinh dfi d u g c phd Kf 18 - 20 logl Wiing slnh k h i c nhau, c i c logi the n i y bao gfim 64 g i l n g vdl nhifiu nflng d f dfi lim g l i trj nflng d f u c chfi t i l t h i l u cda k h i n g sinh d l l vdi vi k h u l n V.'cholerae.

Xd- IJ s l l i f u : Xd-1;? k i t q u i b i n g chuong trlnh Excel; Minitab 14.0.

K t T QUA v A T H A O L U A N

T l I f n h l i m Vibrio cholerae trSn c i c logl m l u p h i h l i p B i n g 2. Tfing bgp H If nhltm Vibrio cholerae tren dc lo^i m i u

TI I f nhiem V. cholerae tren huyet heo d u g c p h i n Igp l i 4 % . Cd nhifiu nghifin c u u cho I h l y n g u l n nu6-c l i y l u t l lay lan m i m b f n h , l i dieu k i f n dfi vi k h u l n ifin tgi v i p h i t l i n . Nhifiu qufic gla t r f n the gidi cung cd nhlj-ng nghiin c u u lifin quan dfin ngufin n u d e g i y 6 nhiSm, nuflc ddng vai trd quan trgng vk l u u tttt mam b f n h , h o n 50% n g u d i bj nhifim vl k h u l n V. cholerae tk do si> d y n g ngufln nu-dc bj fl n h i l m (Joao P.S. Cabral, 2010).

huyfin C l u Naana v i Duwin H i i li, m o i i^Ai „ * * - . . ''^ '^ " ^ ' * T ^- <^f^olerae tren nghfiu d u g c p h i n I f p 6 2

5.93% t4^ Mn i ^ „ a u ' h M b#nh s T p h O T T h ? M % ™ 7 'u»»n M " * - "11° " " l " - * ^ " ^ " " " " ^ ' * " " " " " =

* 4 y n s , a i thS do He rhSn g S , b * h k h i i n h ! , d ™ k h u S „ T » K a t o ^ ' 2 f e = ' ' ' " ^ ""^^ ' ^ " " " " ^ " * " " " " " " " * "

TM« n h i i m WDrio rtoleme M n nghSu * h u y j n C i u Ngang v4 DujrSn Hal

B i n g 1 Tl I f nhlim Vibrio chderae tr«n rtghfiu t?i 2 huyf n

" " > * " Sfimiutalmtra Dirong tlnti Logi miu Sfi mfiu kifin

Huyitheo 100 - Nghflu 160 Phin bfnh nhin 40 ' Tfing

^^

tra Duong tinh

Sfi miu T l l f ( % ) 04 16 0 20

4,0 10,0 0

6,7

_ Qua bang chung tfli n h f n I h l y fi I f d u o n g tfnh v d i V.

Sfi miu Ti 16 (%i Cholerae trfin nghfiu cda h u y f n C l u Ngang l i 1 3 , 7 5 %

^ M i e ( % ) cao hon SO v d i ti I f d u o n g Ifnh v d i V. cholerae t r f i n S ^ Z . S " = " = S ' " ™ ' " " y e n Duyen H i i 14 6.25%, tuy n n i B n i

" " " ' " ^ ° ° " '3.75 * « " ' ' I W nay M n g ca ) nghia ih6ng KS ( P 0.111).

OJc tinh oJa vl khuSn V. cholem 14 thfch nghi v4 t i n HI a u w 6 m6i t r u i n g n u d e co n i n g a i m u i i t h f c h

HOI NGH! KHOA HQC CONG NGHf SINH HQC TOAN Q 0 0 C ^ 0 T 3 ~

-hgp-2%.-3%-vi.4%_vLkhuln h i f n d i f n pho b i l n d bifin. c i c cua song v i c i c l o i i h i i san n h u Iflm, cua, s d , ngheu, h i u

[Vo van L u g n g , 2009], NgudI se bi n h i i m V. cholerae khi i n song h o f c chua n l u chin c i c lo?i"hai san fFgrrhoa(rufihg hay tifip xdc c i c logi nude co nhifim vi k h u l n V. cholerae, n h l t l i n u d e co do m f n t d 2 - 3% NaCI. Cf Khanh Hoa ( V i f t Nam), I d nam 19S7den 1999 06 nhifiu ca nhiem V. chderae, v i K h i n h l-loa cung l i vung ven bien, thdc an s u dgng da sfi l i h i i s i n . Theo b i o c i o t h u d n g nien cda Tmng t i m k i l m s o i l djch bfnh thfi gidi nam 2011 (CDC), t h d c an h i i s i n cdng d u g c chdng minh vfi tl I f n h i l m V.cholerae d nhifiu n u d e frfin thfi gidi: h i u 65"/o, tflm 52%, cua 29%, nghfiu 11 % (DePaola, 1981).

T i I f nhifim Vibrio cholerae tren huyet heo t^l m f t s l h u y f n Bang 4. Tl 1| nhilm Vibrio cholerae trfin huylt heo t^i m f t so Huyfn

Sfi mfiu T i l f ( % ) Duyfin H i i

C l u Ngang Chiu Thinh Cing Long T-PhfiTriVmh

Kfit qua cho thay "tgi c o s d g i l t mfi heo thufc h u y f n C h i u Thanh cfl 2 0 % sfi mfiu d u g c p h i n I f p nhifim vi k h u l n V. diolerae, fi I f n i y cQng k h i cao ' so vfli fi If nhiem da d u g c p h i n I f p trfin nghfiu ct^a c i c huyfn (10%), vdl tt I f nhiSm V. cholerae p h i n , l f p d u ^ c , Chung Ifl nirfl^ cfl k h i nang l i ngufin l i y

n h i l m V. cholerae.

Tfing

Type huy6l thanh Inaba + Ogawa * 0139 Inaba (dan gli) Ogawa (don gli)

sfi mlu duong tinh (n = 20) . Duong tlr^ ' n i f { % )

3 15

^ 10 2 10 Kfit q u i x i c djnh type h u y l t t h a n h p h o bifin g i y b f n h tqt T r i V i n h

B i n g 5. Tl l | nhilm t/ibrio cholerae thao type huylt thanh > _ Kfit q u i cho I h l y cfl 15% m l u duong llnh v d i k h i n g h u y l t thanh da gia (0139, Ogawa, Inaba), 10% m l u duong tinh vdl k h i n g huyfit thanh don g i i Ogawa v i 10% mau duong tinh vdi I d l i n g huylt thanh d o n g i i Inaba.

Tgi Vift nam, t d n a m 1979'dfin 1981 c i c ca b f n h t i chl] y l u l i do biotype El Tor, serotype Ogawa, t d nam 1982 den 1990 c i c ca bfinh ta dfiu n h i l m serotype Inaba;

nhu-ng n i m sau 1990 c i c ca b f n h dfiu d o serotype Ogawa (Joao P.S. Cabral, 2010). T h i n g 10 n i m 2007, c i c trudng hgp lifiu c h i y d p gay ra bdi bien doi gen vi k h u l n Vibrio cholerae 0 1 , Ogawa biotype El Tor v i cfi d i l n (Nguyen ef al, 2002).

T i n h nhgy c i m v i de k h i n g k h i n g s i n h cua v i k h u l n Vibrio cholerae.

ChOng tfli da lifin h i n h t h u k h i n g slnh d l d l d i n h g l i linh nhgy cam cda 20 chdng V. cholerae nghiSn c d u doi vdi c i c logi k h i n g sinh thudng s d dyng dfi difiu trj b f n h la.

Bang 6. Tl If nh^y c i m v i dfi khang khSng slnh doi vdl Vibrio cholerae

Khing slnh Kf hi$u S6 mSu Nhay cam Tmng glan KhSng kiimba ,

S6 mSu {%) Amoxicillin

- Tetracycime Norfloxaclne Chloramphenicol

Azithromycin

Kfit q u i cho t h i y 100% c i c chdng V. cholerae nhgy c i m v d i norfloxadne, 7 0 % vfli chloramphenicol, 65% vdi tetracycline, 50% vdi azithromycin; dfi k h i n g vdi amoxidllin l i 90%, vdi tetracycline l i 30%, vdi azithromycin l i 25%, ket q u i nghifin cuu cda chdng tfli phd hgp vdi nghien cuu cua Nguyfin Th] X u i n Trang efa/., 2012. N i m 1995 vi k h u l n V.

cholerae 0 1 nhgy vdi tetracycline v i diloramphenicol, nhung dfin n i m 2000 vi k h u l n V. cholerae 0 1 Igi d f i k h i n g vdi tetracycline v i chloramphenicol (Vijayalakshmi etal.. 1997) vdi amoxicillin (Bani e( al., 2007).

Cf mifin Bac Vlft Nam t d n i m 2007 dfin 2010, t i t c i chdng V. cholerae p h i n Ifp d u g c dfiu nhgy vdi chloramphenicol (100%), de k h i n g vdi tetracycline l i 2 9 % (Huu Dat Tran e( al, 2012).

HOI NGHI KHOA HOC cONG NGHf SINH HQC T O A N Q U O C 2013

fc:^

B i l u dfi 3.1. Sg nh?y c i m vi d l khing khing slnh c u a c i c c h d n g V. cholerae Nfing d f d c c h l tfil thlfiu (MIC)

B i n g 7. K i t q u i x i c ^ n h nInB d f dc c h l tfil t h i i u cOa khing sinh dfii v d l Wh«o cholerae Lugng Idling

sinh (pg)

' NfngdfMIC (pg/uD

T e l r a c y d ^ Noffloxadna ChkvamphenlGol

AzHhromydn

Qua kfil qua d Bang 6 n h i n I h l y nfing dfl u c chfi t l i t h i l u (MIC) dfii v d i c i c loai k h i n g sinh lan l u g t l i 4 |jg/pl. < 1 PSlyl 0,25 pg/jjl, < 2 (jg/pl v i < 1 . K i t q u i cda nhflm t i c gia Nguyen et al (2002) cOng cho thay MIC doi v d i tetracycline l i 3.^2\sgl\l\. chloramphenicol la 6.25 pg/pl.

K ^ T L U A N

Kfil q u i kifim Ira sg l i y nWfim V. cho/erae trfin 300 mSu ( m i u ngheu, h u y l t heo, p h i n b f n h n h i n ) d T r i Vinh cho t h i y t i n s l x u l t h i f n V. dmlerae trkn nghfiu l i 10%, m l u huyfit heo cfl dha n u d e v d i n i n g d f mufii 2 - 3 % l i 4 % , tren b f n h n h i n Hfiu c h i y chua p h i n I f p dirge V. chderae. Vi Idiuan p h i n I f p nhgy v d i norfloxacine (100%), v d i chloramphenicol (70%), tetracycline (65%). aathromydn (50%), dfi Idling vdi amoxicillin (90%). vdl tetracycline (30%), vdi azithromycin (25%). Nfing d f dc c h l t l i thlfiu (MIC) dfli vfli amoxillin l i 4 pg/pl, tetracycline l i <1 pg/pl, norfloxacine l i 0,25 pg/pl. v d i chloramphenicol l i < 2 pg/pl, vfli aathromydn l i <1 pg/pl.

Cfl 15% m l u duong Unh vdi k h i n g h u y l t thanh da g i i (0139, Ogawa, Inaba),10%'m3u d u o n g h'nh v f l i k h i n g h u y l t thanh don g i i dgawa v i k h i n g huyfit thanh don g i i Inaba.

T A I U $ U THAM K H A O

t<4guyfin Thj XuAn Trang, Nguyin NgpcTuin (2012). T i n sfi kuSt hi$n Vibrio choierae tren tam va nhuyln thfi, x i c <^nh Serogroup 0 1 , 0139 V* Btotype cua V. cbolerae b&ng kJ thuft Multiplex - PCR". Kftos hgc ky thuft ThO y 19(3):51-55

Ph^m ThI VQ (200B): "tighiin cdu ung dyng mflt sfi kJ Ihuf t chin doin nhanh Vibrio cholerae g i y djch bSu chiy c i p tgi tinh Thai Nguyen nim 2008'. Lutfn vin thvcsa SaOi hgr. 29-31

Banl. S., Maatromaiino, P. N . Ceccarolli. D., La Van, A., Salvia, A. M . Ngo Viet. Q T., Hat, D. H., Bacciu, D., Cappuccinelli, P. &

Cokxnbo. M. M. (2007), Molecular charactenzaton of ICEVchVieO and its disappearance in Vibrki chderae 0 1 strains isolated in 2003 bl Vietnam /¥M5MicmbfofLeR266(1).42-S

DePaoia. A {1981). Vibno chderae m marine foods and environmen4ai waters: a literature review. J. Food Sd. 66-70.

Dong Tu Nguyen, Tuan Cuong Ngo, Huy Hoang Tran, Thanh Huong Le, Hoai Thu Nguyen, (2012} Characterizaeon of Vibrio chderae 0139 of an Aquatic Isolate in Northern Vielnam.Open MiaotuolJ, 6:14 - 21

Huu Dat TiOT . Munirui Alam. Nguyen Vu Tmng, Nguyen Van Kinh, Hong Ha Nguyen. Van Ca Pham. Mohammad Ansanizzamart. Shah Mana^Rashed Nunji A. B h u r ^ . Tuyet Trinh Dao, Hubert P. Endtz. and Heiman F. L. Wertheim (2012). Mulli^Jmg resistant Vibrio cholerae 01 variant H Tor isolated n northern Viebiam between 2007 and 2010. Joumd of Medical Mierobidogy. fil: 431-437.

Jww P.S. Cabral {2010).-Water Micmbiotogy. Bacterial Pathogens and Water, int J. Environ. Res. Pudic Health T. 3657-3703.

in Vietnam. Japanese Joumal d

y SIMM, MM (C1.SI. 2010).P.rfO!man«Sl.ndarts lor A n l m t o b l . l S u s c ^ » i l i t y Tasting h i p J M i B c l « > s . l r w fr*iili«ijAhoahocWI«>aJcl«*la htm. V i VSn Urong. 3/8/2009.

HOl NGHI KHOA HQC CONG NGHE SINH HQC TOAN QUOC 2013

" P R E V A t E N C B i 6 ; N D - A N T I B I O f t e - R E S I S - T A N e e ^ M / / e R / e - e H 0 t E R A ^ ISOLATED FROM SWINE BLOOD SAMPLE, CLAM AND PATIENT WITH DIARRHEA IN TRA VINH PROVINCE

Nguyen Thi D a u \ Nguyen Thuy U n h \ Ho Thi Viet Thu^

^Tra Vinh University 'can Tho University

Isolation of Vibrio choleras was carried out on 300 samples including 160 clam samples, lOO swine blood sanqiles and 40 feces of dianfaeal padents collected fiom Tra Vinh hospital, Tra Vinh province.

Results indicated that there were 10% clam samples and 4% swine blood sanqiles contaninated by V. cholerae, whereas no V.

cholerae was isolated from diarrheal feces. The results of kiity bauer antibiotic sensitivity tests showed that all of the isolated V.

chlolerae sCrams were sensitive to norfloxacin (100%), chloramphenicol (70%) but 90% of the isolates is resistant to amoxicillin.

Three out of 20(15%) V. cholerae isolates werepositiveformultivalentantjsera (0139, Ogawa, Inaba), 10% (2/20) of diem ivas positive for monovalent antisera (Ogawa and loaba)

K^ -words: Vibrio choleae, clams, infection rate, antibiotic resistance, Tia'Vinh

•Author for con-espondence: OT: 074.3855246, DD: 0913.671.850; Fax. 074.3855217; Email, ndngiiyandau@gmail.com