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GVHD: PGS.TS TRNH XUN NG

A. GII THIU MT S NT V C QUAN TH Y VNG VI

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Cn c theo quyt nh s 80/2006/QD-BNN ngy 18/09/2006, C Quan Th Y Vng VI trc thuc Cc Th Y c thnh lp trn c s hp nht v t chc li Trung Tm Th Y vng thnh ph H Ch Minh v B Phn thng trc ca Cc Th Y ti thnh ph H Ch Minh. a ch: 521/1 Hong Vn Th, Phng 4, Tn Bnh, Tp H Ch Minh. 1. Chc nng v nhim v, quyn hn: 1.1. Chc nng: C Quan Th Y Vng VI thc hin chnh snh qun l nh nc v th y v chn on, xt nghim bnh ng vt, sn phm ng vt ti vng c phn cng. C Quan Th Y Vng VI c t cch php nhn, c con du ring, c hng kinh ph t ngn sch Nh nc, c m ti khong theo quy nh ca php lut. Tr s ca C Quan Th Y Vng VI t ti thnh ph H Ch Minh. Phm vi hot ng ca C Quan Th Y Vng VI gm cc tnh, thnh ph trc thuc Trung ng (sau y gi chung l tnh): thnh ph H Ch Minh, B Ra Vng Tu, ng Nai, Bnh Thun, Ninh Thun, Bnh Dng, Bnh Phc, Ty Ninh, Long An, Tin Giang v Bn Tre. 1.2. Nhim v, quyn hn: Tham gia xy dng vn bn php lut, k hoch, chng trnh, d n, tiu chun, quy trnh, quy phm k thut v th y. Hng dn, n c, kim tra vic thc hin cc vn bn quy phm php lut, chng trnh d n, tiu chun, quy trnh k thut v th y i vi Chi Cc Th Y cp tnh.

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V phng, chng dch bnh ng vt:

+ Gim st tnh hnh dch bnh; lp bn dch t; xut cc bin php phng chng dch bnh ng vt trong vng. + Phi hp v h tr cc Chi cc th y trong vic chn on, xc nh bnh v t chc vic phng, chng dch bnh ng vt. + Kim tra, chn on, xt nghim nh k bnh ng vt ti cc c s chn nui theo quy nh. + Hng dn v nh gi vic xy dng c s, vng an ton dch bnh ng vt. V kim dch ng vt:

+ Thc hin kim dch ng vt, sn phm ng vt xut khu, nhp khu, tm nhp ti xut, tm xut ti nhp, chuyn ca khu, qu cnh Vit Nam. + Trc tip x l hoc hng dn, gim st vic x l cc cht thi, phng tin vn chuyn, dng c cha ng, ng vt, sn phm ng vt theo quy nh. + Qun l cc Trm Kim dch ng vt ti cc ca khu, ga, sn bay, bn cng, bu in quc t theo s phn cng ca Cc trng Cc Th y. V kim sot git m:

+ Trc tip kim sot git m ng vt ti cc c s git m xut khu trong vng. + Hng dn, kim tra vic thc hin v sinh, kh trng, tiu c dng c bo qun, cha ng, bao gi; x l ng vt, sn phm ng vt khng tiu chun v sinh th y v x l cht thi ti cc c s git m xut khu.
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V kim tra v sinh th y:

+ Kim tra iu kin v sinh th y ni cch ly kim dch ng vt, theo di cch ly kim dch ng vt, sn phm ng vt xut khu, nhp khu theo hng dn ca Cc Th y. + Kim tra iu kin v sinh th y i vi cc c s git m, bo qun, s ch ng vt, sn phm ng vt xut khu, nhp khu v cc c s chn nui theo s phn cng ca Cc trng Cc Th y. + Kim tra v sinh th y ng vt, sn phm ng vt xut khu, nhp khu, chuyn ca khu, tm nhp ti xut, tm xut ti nhp, qu cnh, mn ng Vit Nam. Tham gia tuyn truyn, ph bin kin thc, gio dc php lut, thanh tra chuyn ngnh th y. Tham gia thc hin khuyn nng v v sinh an ton thc phm trong lnh vc th y ti cc tnh trong vng. Tham gia nghin cu, ng dng tin b khoa hc k thut v hp tc quc t theo chng trnh, k hoch ca Cc Th y. Phi hp vi Chi cc Th y cc tnh trong vng t chc o to, bi dng k thut, nghip v v th y. Thc hin thu, np ph v l ph theo quy nh. Qun l t chc, cn b, lao ng, ti chnh, ti sn ca C quan theo quy nh. Thc hin ci cch hnh chnh, chp hnh ch thng tin, bo co nh k, t xut theo quy nh ca Cc Th y. Thc hin nhim v khc do Cc trng Cc Th y phn cng, u quyn theo quy nh ca php lut.
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2. T chc b my: Lnh o C quan Th y vng VI c Gim c v Ph Gim c do Cc trng Cc Th y b nhim, min nhim theo quy nh v phn cp qun l cn b ca B. Gim c chu trch nhim trc Cc trng v trc php lut v hot ng ca C quan v thc hin nhim v c giao. Ph Gim c gip vic Gim c, chu trch nhim trc Gim c v trc php lut v vic thc hin nhim v c Gim c phn cng. B my qun l: + Phng Tng hp. + Phng Kim dch, kim sot git m. + Phng Dch t. Cc n v trc thuc: + Trm Kim dch ng vt xut nhp khu Sn bay Tn Sn Nht. + Trm Kim dch ng vt xut nhp khu Vng Tu. + Trm Kim dch ng vt xut nhp khu Mc Bi. + Trung tm Chn on, xt nghim bnh ng vt.

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S ban lnh o C Quan Th Y Vng VI v cc trm, n v trc thuc:

Phng Tng Hp

Cc trm kim dch

Kim dch v kim Sot git m

Trung tm chn on v xt nghim bnh ng vt

Dch t

Trm sn bay Tn Sn Nht

Trm Mc Bi Ban Gim c TTCD - XN Ban qun l cht lng

Trm Tn Cng

Trm Vng Tu

T Tng hp

virus

Vi trng

Ho l

Bnh l

X l dng c v mi trng

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B. BO CO TI

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PHN I. M U

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1.1.

T VN An Ton V Sinh Thc Phm l mt vn ngy cng c cp v quan tm

hiu hn. Trong , ng c thc phm ang l mi lo ngi rt ln ca cng ng v gy nh hng ti sc khe, lin quan n nhiu loi bnh khc nhau. Trong , bnh vim d dy- rut c xem l ph bin nht trong s cc bnh do ng c thc phm, bnh c gy ra bi vi khun, virus v cc k sinh trng khc. Trong s cc vi khun gy ng c hoc gy bnh qua thc phm, Salmonella l i tng gy ng c ph bin nht. Thc n hoc ngun nc nhim Salmonella c th gy nn nhiu bnh nh vim d dy rut, thng hn v nhim trng mu. Hu ht cc trng hp ng c do Salmonella u khng gy t vong. Tuy nhin tr em, ngi gi v bnh nhn mi phc hi t bnh trng khc cng c th nguy him tnh mng khi b ng c Salmonella. Mt s serotype (kiu huyt thanh) ca Samonella chuyn bit vi tng vt ch, v d nh Samonella choleraesuis ch gy bnh trn heo, S.abortusovis ch gy bnh trn cu, S.pullonella ch gy bnh trn gia cm, Tuy nhin, phn ln cc serotype khc c th gy bnh ph bin rng khp cc vt ch, k c ngi. Do vy, ngoi tr mt s serotype gy nhim bnh chuyn bit trn mi loi ng vt, cn khong 2500 serotype khc u c xem l c th gy bnh cho ngi. T nhng nhn thc trn cng vi s hng dn ca thy PGS.TS Trnh Xun Ng- Vin Cng Ngh Sinh Hc & Thc Phm trng i hc Cng nghip Thnh Ph H Ch Minh v c s ng ca C Quan Th Y Vng VI, ti thc hin kho thc tp tt nghip vi ti: KHO ST T L NHIM SALMONELLA TRONG SN PHM THT G NG LNH NHP KHU.

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1.2.

MC CH Kho st t l nhim Samonella trn sn phm tht g ng lnh nhp khu.

1.3.

YU CU Tm hiu v nm vng quy trnh phn tch ch tiu Samonella trong tht ng lnh

nhp khu ni chung. c bit l sn phm tht g ng lnh nhp khu ni ring.

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PHN II. TNG QUAN V TI THC TP

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2.1.

S LC V VI KHUN SALMONELLA

Vi khun Salmonella thuc b Eubacteriales, h vi khun ng rut (Enterobacteriacae). Vi khun Salmonellac tr ch yu trong rut nn ngi ta gi Salmonella l trc khun ng rut. Vo nm 1935, chng vi khun Salmonella l u tin c Salmon v Smith tm thy l Salmonella choleraesuis. k nim ngi u tin tm ra vi khun ny, theo ngh ca Hi ngh Vi sinh vt hc quc t, ngi ta t tn cho vi khun ny l Salmonella. Vi khun Salmonella l trc khun gram m, hnh gy ngn, kch thc t 0.40.6x1-3, khng hnh thnh gip m v nha bo, hiu kh ty tin. a s cc loi Salmonella u c kh nng di ng do c long roi xung quanh thn (t 7-12 lng) (tr Salmonella gallinarum v Salmonella pollorum). Vi khun Salmonella c th pht trin c mt s loi mi trng thng thng v mt s mi trng c hiu nh mi trng BHI(brain heart infusion), mi trng thch mu, mi trng Tetrathionat, XLD agar, MacConkey agar. Trong qu trnh nui cy, Salmonella c th bin d v khun lc v khng nguyn. Vi khun c th bin d khun lc t dng S (Smooth) sang dng R (Rough). (Nguyn Nh Thnh v cng s, 1997). Cc tc gi cn cho bit: vi khun mi phn lp c dng S, c khng nguyn O c hiu ca chng nhng qua mt thi gian, vi khun bin d khun lc, t dng S sang dng R v khng nguyn O khng cn c hiu na. Ngoi ra, di nh hng ca mt s cht nh acid phenic, vi khun s mt long v gy bin d (Nguyn Nh Thnh v cng s, 1997) Trong mi trng thch mu, vi khun khng gy dung huyt v trong mi trng gelatin, vi khun khng lm tan chy gelatin. Phn ln cc loi Salmonella ln men v sinh hicc loi ng nh: Glucose, Maltose, Mannitol, Galactose, Levulose v Arabinose. Salmonella kng phn gii ure, khng sn sinh indole, sn sinh H2S (tr Salmonellaparatyphy A, Salmonellaabortus equi, Salmonellatyphy suis) a s vi khun Salmonella cho phn ng Catalase, MR dng tnh, trng hp ngoi l Salmonella choleraesuis, Salmonellagallinarum, Salmonellapullorum cho phn ng MR m tnh. C khong 96% cc chng vi khun Salmonella tit ra enzyme kh cacboxyl i vi lysine. Tt c cc chng Salmonella u khng ln men ng Lactose, Saccharose.
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Theo phn loi ca Kauffmann- White, gng Salmonella bao gm mt chng n l c phn chia thnh 2000 serotypes da trn khng nguyn thn (O), khng nguyn roi (H) v khng nguyn v thng thng (Vi-anigens). Trong nhng nm gn y, ging Salmonella c chia lm 7 nhm ph (Subgroups). Nhm ph I gm hu ht nhng vi khun Salmonella m c kh nng gy bnh cho ng vt v phn ln chng c c tn nh Salmonelladublin hay S. typhimurium. Nhm ph IIIa v IIIb gm loi vi khun m ch c bit nh l Arizona v ngy nay c gi l Arizonaenu pha n (IIIa) hay Diarizonae nu pha i (IIIb). C gn 400 serotypes trong nhm ph IIIa v IIIb. thun tin cho vic chn on phng th nghim v cch gi tn, ngi ta a ra mt danh php c n gin ha m t tnserotypes caSalmonella l loi (species), v d nh Salmonelladublin hay Salmonellatyphimutium. Ngay trong mi loi ngi ta cn phn ra nhiu serovar hay serotypes da vo tnh cht khng nguyn thn (khng nguyn O), v khng nguyn roi (khng nguyn H). Qua cc cng trnh nghin cu, ngi ta thy rng hu ht cc loi Salmonella c bit u c kh nng gy bnh cho ngi hoc ng vt, c loi gy bnh cho c ngi v ng vt. 2.1.1. x Salmonella O ) 2339 serotype ( (

Enteritidis (S.enteritidis), Typhi (S.typhi), Paratyphi (S.paratyphi), Typhimurium (S.typhimurium) . Salmonella , Salmonella . Salmonella c chia thnh 2 chng:
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yu theo

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- S.enterica: c 6 loi ph: + S.enterica I + S.enterica II + S.enterica IIIa +S.enterica IIIb + S.enterica IV + S. bongori (Salmonella V) + S.entericaVI Salmonella thng c gi tn theo kiu huyt thanh: S. typhi, S.enteritidis, S. TyphimuriumCc kiu huyt thanh gy bnh ch yu nm trong S.enterica I. Salmonella - S.typhi (trc khu . - S.paratyphi A S.typhi. - S.paratyphi B . Chng l cn nguyn gy ra bnh ph thng hn. - S.paratyphi C . - S.enteritidis (trc khun Gartner, 1888) v S.typhimurium (trc khu :

Salmonella.
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- S.choleraesuis monella .

t do Sal

... Mt s Salmonella khc: S.thompson, S.derby, S.newport, S.kissangani,

S.senflenberg, S.meleagnidis, S.anatum, S.aberdeeh, S.panama, S.montevideo, S.dublin, S.london, S.anatum, S.minnesota, S.gallinarum 2.1.2. Cu to khng nguyn ca vi khun Salmonella Cu to khng nguyn ca Salmonella rt phc tp. Salmonella, ngoi phn ng huyt thanh c hiu ca tng vi khun cn c hin tng ngng kt cho gia khng nguyn ca vi khun ny vi khng th ca vi khun khc hay gia nhm ny vi nhm khc trong ging. c hin tng trn t ngng kt hay ngng kt cho gia cc loi vi nhauxy ra do vi khun Salmonella c nhng thnh phn khng nguyn c hiu hay khng c hiu chung cho nhm. Cu trc khng nguyn ca Salmonellagm c 3 loi: khng nguyn O, khng nguyn H v khng nguyn K (khng nguyn Vi-anigens). Khng nguyn O: Kauffmann v White nghin cu mt cch h thng cu trc khng nguyn O ca Salmonella. Khng nguyn O ca vi khun Salmonella rt phc tp, cu trc khng nguyn gm nghiu phn t cu to nn v n c phn b trn b mt ca t bo. Thnh phn ch yu ca khng nguyn O l phospholypid, polysaccharide, trong c 60% l polysaccharide, 20-30% l lipid v 3.5-4.5% l hecsosamin. Kauffmann v White ch ra gn 70 yu t khng nguyn khc nhau. Mi loi Salmonella c mt hoc mt s yu t trong s cc yu , mi yu t c k hiu bng cc s la m hay s rp.
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Do c s khc nhau gia cc loi Salmonella v cu trc khng nguyn O nn ngi ta chia Salmonella thnh 34 nhm v c k hiu bng cc ch in A, B, C1, C2, C3, D1, D2,, X, Y, Z. Mi nhm vi khun c khng nguyn O cu to bi mt s thnh phn nht nh c k hiu bng s La m. Khng nguyn H: Khng nguyn H ch c nhng vi khun Salmonella c lng nn hu ht Salmonella u c khng nguyn H (tr Salmonella gallirum v Salmonella pullorum). Khng nguyn H chia lm 2 pha, pha 1 c tnh c hiu gm 28 loi khng nguyn lng c biu th bng ch la tinh thng a, b, c, d, f, g, h, , z; pha 2 khng c tnh c hiu v gm 6 loi c biu th bng ch s rp 1, 2, 3, 4, 5, 6 hay ch la tinh thng e, n, x, V vy, ngi ta thng dng cng thc khng nguyn O, khng nguyn H gi tn cho loi Salmonella. Khng nguyn H khng chu nhit, b nhit 70oC ph hy nhng khng vi foocmol. Khng nguyn K: Khng nguyn K ca Salmonella khng phc tp v ch c S.typhi v S.paratyphi, c gi l khng nguyn Vi (virulence). Khng nguyn Vi di dng mt lp rt mng khng quan st c bng knh hin vi quang hc thng thng. khng nguyn Vi c bn cht l mt phc hp glucid-lipid-polypeptide gn ging khng nguyn O nhng s khng ngng kt mc d trn vi khng nguyn O c hiu. hin tng ngng kt xut hin ngi ta un nng dch vi khun 100oC/20 pht tch khng nguyn K ra khi t bo. Khng nguyn Vi gp khng th Vi xy ra hin tng ngng kt chm v xut hin cc ht nh. Khng nguyn khng tham gia vo qu trnh gy bnh 2.1.3. Salmonella , trung ,h .

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Cc sn phm tht ni chung, nht l tht gia cm v tht ln,tt c cc thc n ti sng c ngun gc ng vt u c th l ngun ly nhim vi khun Salmonella. Vi khun ny sng t do trong rut ng vt. Gia cm c nhiu Salmonella nht, tip theo l cc ng vt nui trong nh v ng vt hoang d (vt, ra, ch, ch, chim mng bin, loi gm nhm, rn). Vi khun ny c th c trong thnh phn dn xut cc cht t ng vt nh gelatin hoc nc bt ng vt, bi cn trng, loi gm nhm, chim hoc cc sn phm tht nhim khun gy nhim vo thc phm. Ngoi ra c th b nhim Salmonellat ngi khe mnh c mang vi khu . Bi v b c ch bi pH < 4 v c mt vi khun lactic nn cc sn phm ln men t b nhim. Trng v cc sn phm trng, v d nh bt nho, nc st mayonnaise, protit ng t tch t sa cng l ngun c th b nhim Salmonella. Bi v, gia cm l ngun mang nhiu Salmonella, nn trng ca n cng rt d b nhim.Nguyn nhn l do vi khun ny c th xuyn qua v trng v pht trin trong lng trng. Trong trng hp ny, khng p dng c phng php thanh trng Pasteur thanh trng vi khun. Do , i vi nhng mn trn vi mayonaire cn phi c kim tra k hn. Cc sn phm sa nh sa khng thanh trng, phomt t sa ti, kem, cht bo sa v cc sn phm t sa ni chung c ch bin t cc nng tri, cc thit b dng c u c th b nhim Salmonella. T , chng gy nhim c cho sn phm sa. Nu tin hnh axit ha chm th vi khun d dng sinh sn trong phomt nhng n b ph hy vi pH < 4,5. Nhng sn phm c sa phi c gim st cht ch bi chng khng c thanh trng na, v vy nu c Salmonella trong sa bt th chng vn c th sinh sn c bi chng c kh nng tn ti iu kin kh v ly nhim sang cc sn phm khc. i vi cacao v chocolate, cacao c rang cn socola c thanh trng Pasteur tiu dit Salmonella, cng c th sau khi thanh trng cn st Salmonella trong socola. C trng hp nhim t cht bo ca sa nhng phn ln l t trng ti b sung vo cc n.
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cc nc ang pht trin, nn sn xut nng nghip cn lc hu, nc b nhim Salmonella bi do ri phn sc vt v cc bin ti sng hoc cha chn, c bit l ng vt thn mm, 2 mnh v. Trc kia,loi Salmonella typhi nhim nhiu nht vo ng vt thn mm, nc khng qua x l Clo, hay phn ca ngi b bnh thng hn. Hoa qu v rau xanh: c th b nhim khi ra bng nc b nhim khun hoc bi tay ngi ra mang vi khun. Salmonella sinh sn tt trong qu da v c th chu c nhit khi thanh trng pasteur khng y hoc b nhim sau khi thanh trng, hay c th b nhim t nc to khng thanh trng. Sn phm ng cc nh: u tng, ng cc ch bin v cc mn khai v, bnh m, bt u tng, c linh lng c th l ngun Salmonella do b nhim t phn bn c cha Salmonellatrong qu trnh trng trt v khi sn xut m ngm trong thi gian di nhit thng. Nu nhng sn phm ny c dng trc tip m khng x l nhit th ngi s dng s c nguy c b nhim Salmonella. Trng hp ng cc c ch bin v cc mn khai v, s c th b nhim Salmonellat nguyn liu nh: trng, sa, vitamin, cht ngt v cht to mu. Cc thc phm n nhanh nh loi bnh c tht, sa hoc trng, c th b nhim khun, nht l nhit bnh thng. Khi t b Salmonella teinth . 2.1.4. c lc v triu chng lm sng ca bnh do Samonella Cc th nghim cho thy liu gy bnh ca Samonella t 105 n 107 t bo (McCullough v Eisele, 1951). Tuy nhin, theo kho st t cc ca nhim thc t , liu gy bnh ch c th t 10- 45 t bo (DAoust v cng s, 1985; Lehemacher v cng s, 1995). Nhiu bo co chng minh liu gy bnh ca Samonella s thp hn khi n hin

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din trong thc phm c nng cao ca protein v cht bo, nhng c cht gip bo v vi khun khi pH acid ca dch v (DAoust v cng s, 1975; Blaser v Newman, 1982) Tu thuc vo c tnh ca tng chng v tu th trng ca ngi b nhim, liu gy bnh ca Samonella s khc nhau, ng thi kh nng v trng thi gy bnh cng khc nhau: t vim d dy- rut n thng hn hay nhim trng mu. Nhn chung, bnh do ng c Samonella ch yu l vim d dy- rut. Thi gian bnh 6- 46 gi sau khi tiu ha thc n, v phn ln thng trong khong 12- 36 gi. Cc du hiu u tin l bnh nhn thy bun nn, nhc u, chong vng, kh chu, thn nhit tng ln t (38-39oC), sau c biu hin vim d dy rut cp tnh: nn ma, tiu chy nhiu ln, phn ton nc, i khi c mu. a s bnh nhn tr li trng thi bnh thng sau mt n hai ngy, mt s trng hp ko di n 7 ngy. Trng hp t vong him khi xy ra, ngoi tr ngi gi, tr em v ngi va phc hi sc khe sau khi mc bnh khc. Trong s cc bnh do Samonella gy ra, bnh thng hn v ph thng hn, do S. Paratyphy gy ra, l bnh nghiem trng nht v c t l t vong cao hn c. Biu hin ban u ca bnh c th tng t vim d dy-rut, ko di trong 1-2 ngy. Sau , cc triu chng khc ca bnh khng chuyn bit, c th bao gm nhc u, st cao, mt mi ton thn, bin n, au vng tht lng v c bp. Nu khng kp thi iu tr, bnh c th dn n vim mng no v nhim trng mu t gy t vong. Ngay sau khi cc triu chng bnh bin mt, vi khun Samonella vn c th tn ti trong rut v trong phn c ngi bnh trong mt thi gian di v vy cn c bin php x l thch hp.

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2.2.

TNH HNH NG C THC PHM DO SALMONELLA

2.2.1. Trn th gii Salmonella l mt trong nhng tc nhn gy bnh qua thc phm ph bin nht trn th gii (Thom v cng s, 2000). Theo s liu thng k ca Thoms (2000), trong nm 1997, s trng hp nhim Salmonella ln n 100.000 ngi, xy ra mt s nc nh: 14 ca M, 38 ca c, 73 ca Nht Bn, cc nc chu u: 16 ca H Lan n 120 ca c. Mt thng k t nm 1995 n 1998 ti khu vc nam M Latinh cho thy s v ng c do Salmonella chim phn ln (36,8%) trong tng s v (Franco v cng s, 2002). Ng c thc phm do Salmonella chim 55,1% trong tng s ca nhim ng c thc phm do vi sinh gy ra t nm 1993 n 1996 Hn Quc (Baik and Roh, 19980). Biu 2.1. T l phn b cc loi Salmonella trn th gii

(Ngun: WHO Global Salmonella Surveillance (2000-2004))

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Mt s loi Salmonella.ssp ch gy bnh ch yu mt s khu vc hay quc gia. Tuy nhin, c loi hin din khp ni trn th gii l S.enteritidis v S.typhimurium. Theo bo co ca chng trnh gim st Salmonella ton cu ca WHO (WHO Global Salmonella Surveillance), thng k s liu ca 75 nc t nm 2000- 2004, S.enteritidis l kiu huyt than ph bin nht trn th gii (61%), tip theo l S.typhimuriumi (18%). Ti chu , S.enteritidis cng chim t l cao nht (33%), k n l S.typhimuriumi (5%) v S.rissen (5%). 2.2.2. Ti Vit Nam Theo kho st ca Cc th y v tnh trng nhim vi sinh vt trong tht, sa ti mt s im git m, ch ca hng kinh doanh H Ni v TP. H Ch Minh cho thy tnh hnh nhim vi sinh vt tht v sa l ng lo ngi. T l mu tht (g, b, ln) khng t tiu chun v c 4 ch tiu vi sinh vt (E.coli, Coliforms, Salmonella, Clostridium) a bn H Ni l 81,3%, c bit tht b l 100% s mu; a bn TP. H Ch Minh l 32%. y chnh l mt trong nhng ngun gy ng c thc phm quan trng. Theo thng k, mi nm Vit Nam c chng 250-500 v ng c thc phm vi 7.000-10.000 bnh nhn, trong 100-200 ca t vong. S v ng c do thc phm b nhim vi sinh vt chim 33-49%, ch yu do cc chng Salmonella, E.coli, Clostridium perfringens, Listeria. Vi khun Salmonella l nguyn nhn ca 70% v ng c vSalmonella tn ti trong nhiu loi thc phm nh: ngui, tht ngui, nghu s, g cha nu chn, ch phm t sa sng, thc n ch bin sn, nht l trong cc mn n ch bin t trng ti hoc cn hi ti sng. Tng t nh trn th gii, S.typhimurium v S. enteritidis cng l loi gy ng c thc phm ph bin nht Vit Nam. Mt bo co da trn cc mu phn lp t cc bnh nhn b tiu chy v st ti mt s bnh vin v ti vin Pasteur TP. H Ch Minh cho thy cc kiu huyt thanh ca Salmonella c pht hin nhiu nht l S.typhimurium (37,5%), S. enteritidis (12,5%) v S.weltevreden (7,1%) (VoT.T.A v cng s, 2005)
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2.3.

QUY TRNH KIM NGHIM

2.3.1. Phng php trch mu, ly mu th nghim Ly mu kim nghim l khu u tin v rt quan trng trong cng tc kim nghim thc phm. Trn thc t ch ly mt lng mu rt nh kim nghim m kt qu li dng nh gi mt cch khch quan cht lng ca mt lng ln sn phm. Vic ly mu ng quy cch s gp phn cho kt qu kim nghim chnh xc v x l sn phm ng n. Tu thuc vo c quan qun l k thut ch bin bin thc phm, c quan qun l cht lng v sinh an ton thc phm, tho thun gia cc bn lin quan v c tnh ca mi loi thc phm m s c yu cu v cch ly mu khc nhau. Tuy nhin v nguyn tc chung, vic ly mu phi thc hin theo nhng quy nh sau y: Mu phi i din cho c l hng thc phm ng nht. Khi lng mu phi tin hnh kim tra cc ch tiu ca sn phm. Cc dng c ly mu v bo qun mu phi c v trng trnh s tp nhim vi sinh vt ngoi lai. Nhng mu tht ng lnh m cha s dng ngay phi c bo qun nhit -

20oC cho n khi phn tch. Trng hp khng th bo qun ng th bo qun -4.4oC trnh s bin i thnh phn c tnh ca mu cng nh ca vi sinh vt.Nhn chung mu phi c xt nghim trong vng 36 gi, nhng mu khng c ng lnh cn xt nghim trong vong 6 gi. lu hn c th tang hoc gim s lng vi sinh vt. Vi nhng vi sinh vt d b tn thng, trong nhiu trng hp cn giai on tng sinh. Ghi chp tn mu, thi gian ly mu, thng tin y , chnh xc.

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Khi ly mu ban u cn ch n trng thi, tnh cht ca sn phm. Phi ly

mu nhiu v tr khc nhau trong c l hang v trong c n v cha c th nh sau: + i vi sn phm th rn cn ch n s khng ng u v kch thc ca sn phm, phi ly c sn phm c kch thc ln v c sn phm c kch thc nh. + Thng ta phi chia im ly mu theo hnh dng ca n v cha sn phm.

Hnh 2.1: Chia im ly mu sn phm th rn.

Khi ly mu cc v tr chia ta cn phi ch n vic ly mu cc su

khc nhau ca sn phm. i vi cc sn phm th lng cn ch : + Nu ly mu lng trn ng ng vn chuyn th thng ta ly vi chy ra. M vi cho cht lng chy ra mt thi gian (2-3 pht) ri mi hng ly mu. Khi ly cng phi ly nhiu thi gian khc nhau. + i vi sn phm va th rn va th lng khng ng nht, ta ly mu nhiu v tr khc nhau, nhng phi li ly c phn rn, c phn lng theo dung t l ca chng trong sn phm. + i vi sn phm ng nht dng st cn khoy k v ly mu nhiu v tr khc nhau. + i vi sn phm c ng trong chai, l, hp hay l bao gi trong ti th ly c ti.
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Ch : dng c ng mu c th l bao b ban u ca sn phm hoc c ng

gi trong cc dng c khng c lm nh hng n sn phm. sng. 2.3.2. Tip nhn mu ti phng th nghim Kim tra trng thi ca mu trc khi tip nhn, nu trng thi khng m bo nh mu b nt b, mu thc phm b dp nt hay khi lng khng th khng tip nhn. Kim tra thng tin ca cc mu ghi trn nhn (ngy ly mu, tn mu) Khi vn chuyn mu n phng th nghim cn phn tch ngay. Khi cha phn tch ngay c cn bo qun mu trong t lnh v trnh bn t mi trng bn ngoi. 2.3.3. Bo qun mu Mu lng, thc phm thng b bin i cc mc khc nhau do cc tc ng l ho, vi sinh vt xy ra trong qu trnh trch mu cho n khi phn tch. Trong sut qu trnh trch mu, vn chuyn v bo qun mu cn m bo mu t b bin i nht. 2.3.4. Quy trnh ly mu ti phng th nghim i vi mu lng Ht 25ml (tng ng vi 25g i vi mu rn), lc k trc khi ht. i vi mu rn Mu tht: dng dao, ko ct cc v tr chia nh trn sao cho lng mu thu c l 25g cho vo ti v trng pha long v ng nht mu. Vi mu gia cm: ct ly ngu nhin trn cc ming tht cc b phn khc nhau sao cho lng mu thu c l 25g
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Thc phm d b h hng: nguyn liu tht, rau qu, cc loi thu hi sn ti

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Vi mu thu sn cng tng t. Ch trc khi ly mu cn v sinh dng c bng cn 70o. X l mu V sinh sch s khu vc lm vic, trnh nhim khun vo mu, cc thao tc ct mu, pha long mu v ng nht mu cn m bo iu kin v trng. Chun b dung dch pha long: Buffer peptone water, i vi mu sa v cc sn phm t sa th chun b dung dch Fraeser c km theo cht b sung. 2.4. QUY TRNH XC NH SALMONELLA TRONG THC PHM Quy trnh xc nh c thc hin theo ISO 6579:2002 Vi sinh vt trong thc phm v thc n chn nui - Phng php pht hin Salmonella trn a thch. y l phng php nui cy nh danh, c gi l phng php truyn thng v p dng trong tt c cc phng kim nghim vi sinh. Phng php cho php pht hin Salmonella trong cc sn phm, thc phm dng cho ngi, thc n gia sc v cc mu mi trng trong ch bin v sn xut thc phm. 2.5. CH TIU XT NGHIM I VI SN PHM THT G NG LNH Theo quy nh ca B y t, sn phm tht g ng lnh thuc sn phm tht ng lnh nguyn con hoc ct ming. Nhm sn phm ny gm 6 ch tiu vi sinh cn c kim tra vi gii hn cho php cn c vo Q s 46/2007/Q-BYT,ngy 19/12/2007 ca B Y t (bng 4.1).

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Bng 4.1.Gii hn cho php cn c vo Q s 46/2007/Q-BYT, (ngy 19/12/2007 ca B Y t)

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PHN III: NI DUNG V PHNG PHP

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3.1.

Thi gian v a im thc tp

Thi gian thc tp: t 02/01-31/03/2012 Ti a im: 521/1 Hong Vn Th, Phng 4, Qun Tn Bnh, TP. H Ch Minh. Phng xt nghim vi sinh thc phm - T vi Trng- TTC Xt Nghim Bnh ng Vt - C Quan Th Y Vng VI. 3.2. Thit b v nguyn vt liu

3.2.1. Thit b T thao tc v trng T m B n nhit L vi sng T lnh My ng nht mu Ni hp Cn k thut Ko Pipt thu tinh 10ml Bp cao su Micro pipt (100-1000ml) a petri ng nghim Lu cc dng c v thip b k trn phi tuyt i v trng. Phng vi trng cng c v sinh sch s, hn ch ra vo trnh tp nhim. Mi trng tin tng sinh: Buffer Peptone Water (BPW) Mi trng tng sinh chn lc: Rappaport Vassiliadis (RVS), Muller-Kauffmann tetrathionate/novobiocin broth (MKTTn)

3.2.2. Nguyn vt liu -

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Mi trng phn lp: Xyclose Lysine Desoxycholate Agar (XLD), Hektoen enteric agar (HE) Mt s mi trng th sinh ha: Triple sugar iron agar (TSI), MR-VP medium, Urease agar, Nutrient agar, L-Lysine decarboxylation medium Mt s thuc th cho cc phn ng sinh ha: Nc mui sinh l, thuc th Kovacs, dung dch Creatine, KOH 40%, - naphtol (1- naphtol), a giy O.N.P.G Hoc s dng kit th sinh ha nh danh cc trc khun Gram (-) vi BIS 14 GNE (Cng ty Nam Khoa).

3.3.

Ngun mu Cc mu tht g ng lnh nhp khu c cc Trm Cng nhn v gi v phng

xt nghim Vi sinh thc phm ca Trung Tm Chn on Xt Nghim Bnh ng Vt thuc C Quan Th Y Vng VI. 3.4. Ni dung nghin cu Xc nh t l cc loi mu tht g ng lnh nhp khu T l nhim Salmonella trong sn phm tht g ng lnh nhp khu

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3.5.

S quy trnh xt nghim Salmonella theo ISO 6579:2002

3.6.

Phng php tin hnh

3.6.1. Chun b mu v dch nui cy Chun b mu v dch nui cy ban u theo ISO 6887-1, ISO 6887-2, 6887-3, ISO 6887-4 v ISO 8216 ty theo tng loi sn phm.

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Thng thng cn 25 g mu c ng nht cho vo bao nylon v trng v b sung 225 ml mi trng buffered peptone water. Nu s dng lng mu khng phi l 25 g th b sung mi trng tin tng sinh (buffered peptone water) sao cho dung dch mu thu c c pha long l 1/10. Do Salmonella thng hin din vi s lng thp trong thc phm, i khi b tn thng v c th tn ti cng vi nhiu vi khun khc thuc h Enterobacteriacea, nn vic phn tch nh tnh vi khun ny trong mu thc phm cn phi qua cc bc tin tng sinh, tng sinh chn lc trc khi nui cy phn lp v khng nh bng sinh ha. 3.6.2. Tin tng sinh Cn v trng 25 g mu th c (hoc 25 ml mu th lng) trong bao dp mu v trng. B sung 225 ml nc pepton m (BPW). ng nht mu bng my dp mu, thi gian 30 giy v trong t m 37oC 1oC trong 18h 2h. 3.6.3. Tng sinh chn lc Cy chuyn 0,1 ml canh pepton vo ng nghim c cha 10 ml mi trng Rappaport Vassiliadis (RVS) v trong t m 41,5oC 1oC trong 24h 3h. Cy chuyn 1 ml canh pepton vo ng nghim c cha 10 ml mi trng MKTTn v trong t m 37oC 1oC trong 24h 3h. 3.6.4. Phn lp T canh trng RVS v MKTTn mc 3.4.3.3, cy phn lp sang 2 mi trng: Xylose lysine desoxycholate agar (XLD) v Hektoen enteric agar (HE) hoc Bismuth sulfite agar (BS) Lt ngc cc a trn v (371)0C/ (243)h cho XLD, 350C/(18-24)h cho HE hoc 350C/48h cho BS Quan st cc khun lc c trng:
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Trn XLD: Salmonella cho khun lc c hoc khng c tm en, c qung trong sng, hi (Ch : nu ln men lactose (+) khun lc s c mu vng) Trn HE: khun lc c mu xanh dng n xanh l, c hoc khng c tm Trn BS: khun lc c tm en, vin sng bao quanh bi vng ta c nh kim. Ly t nht 1 khun lc in hnh t mi trng phn lp, ly thm 4 khun lc nghi ng khc nu khun lc u tin m tnh. Nu t hn 5 khun lc c trng hoc nghi ng th ly tt c khun lc c trng hoc nghi ng th khng nh. Cy ria cc khun lc trn mi trng NA ( lm kh b mt) sau cho thu c cc khun lc ri cc a (371)0C/ (243)h Trn TSI (Glucose dng tnh, sinh hoc khng sinh H2S)

3.6.5. Chn khun lc khng nh -

3.6.6. Khng nh sinh ha 3.6.6.1. Cy cc khun lc nghi ng t NA ln mi trng TSI (cy ria phn nghin v cy cc ng nghim (371)0C/ (243)h Trn TSI Salmonella cho: o Phn ng kim phn nghin (mu ): lactose v sucrose m tnh. o Phn ng acid phn y(mu vng): glucose dng tnh, sinh hi (khong 90%) v H2S (thch c mu en). Khng kt lun ngay l mu b nhim Salmonella nu phn nghing trn TSI c mu vng (lactose dng tnh) 3.6.6.2. Urea agar Cy ria cc khun lc nghi ng trn t mi trng NA vo phn nghing ng cc ng nghim ny (371)0C/ (243)h. Salmonella cho phn ng Urea (-):
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m su phn y) -

nghim cha Urea agar -

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o Phn ng (+): Mi trng chuyn sang mu hng ti o Phn ng (-): khng lm i mu mi trng. 3.6.6.3. L-Lysine decarboxylation medium Cy khun lc nghi ng t mi trng NA vo mi trng L-Lysine (371)0C/ (243)h. Salmonella s cho phn ng (+)(tr S.paratyph A(-)) o Phn ng (+): Mi trng c v c mu tm m hn ban u. o Phn ng (-): Mi trng mu vng. 3.6.6.4. Pht hin -galactosidae: (s dng a giy O.N.P.G) Cy mt vng sinh khi vi khun ly t khun lc nghi ng trn mi trng NA vo ng nghim cha 0,5 mL nc ct. Cho a giy O.N.P.G vo ng ng (371)0C/ (243)h trong b iu nhit. Salmonella s cho phn ng vi -galactosidase(-) o Phn ng (+) mi trng c mu vng o Phn ng (-) mi trng khng i mu. 3.6.6.5. Phn ng VP Cy khun lc nghi ng vo ng nghim cha 3mL mi trng VP ng (371)0C/ (243)h. Nh vo ng canh trng vi khun 2 git dung dch Creatine, 3 git 1-naphtol pha

decarboxylation medium -

trong cn v 2 git dung dch KOH 40%. Lc u mi trng sau mi ln b sung thuc th. Salmonella cho phn ng VP (-) o Nu phn ng (+): mi trng c mu hng trong 15 pht. 3.6.6.6. Phn ng indol Cy khun lc nghi ng vo ng nghim cha 5mL mi trng tryptone hoc tryptophane.
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ng (371)0C/ (243)h Nh 1mL thuc th Kovacs Salmonella cho phn ng Indol (-)(mi trng c vng mu vng trn b mt). o Nu phn ng (+): mi trng c vng mu trn b mt.

3.6.7. c kt qu Salmonella l trc khun Gram m, hiu kh, khng sinh bo t, c kh nng di ng, khng sinh Indol hoc Acetoin, khng ln men Sucrose, Lactose, khng phn gii Urea. kim tra nh tnh Salmonella, ngha l ch xc nh n ging, khng cn n loi. Kt qu y ch cn da trn mt s phn ng sinh ha ph hp l hon ton i n kt lun khng nh. Trong trng hp ny cc phn ng huyt thanh ch mang tnh cht h tr. Bng 3.2. Tng hp c kt qu phn ng sinh ha Salmonella theo ISO 6579:2002 Salmonella Stt Th nghim m tnh (-) Dng tnh (+) (+) Khng xut hin mu xut hin mu 1 Indole m tnh b mt b mt 2 Ure Khng thay i mu cnh sen m tnh 3 Citrate Khng thay i mu Xanh dng mu -hng nht trn 4 Khng thay i mu m tnh VP b mt mi trng Mi trng c mu 5 -galactosidase Khng thay i mu m tnh vng Mu xut hin dc Mu xut hin khp 6 Di ng theo ng cy mi trng 7 TSI
Nghin: (lactose v/hay sucrose m tnh); Su: vng (glucose dng tnh), sinh hi, ta en (H2S +)

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PHN IV: KT QU V THO LUN

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Trong kho st ny, phng php ch kim tra khng nh n ging (Salmonella spp.) nn pht hin bng cc phn ng sinh ha ph hp. Kt qu bo co l pht hin hoc khng pht hin Salmonella spp. trong lng mu c kim tra. 4.1. T l cc loi mu tht g kho st Bng 4.3: Bng theo di s mu tht g ng lnh nhp khu (Thng 3 nm 2012)

Da vo bng 4.3 s liu thng k cho thy mu tht g ng lnh nhp khu trong thng 3 (t ngy 1-25/03) l tng i nhiu 167 mu. Trong , sn phm chim t l cao nht l loi sn phm i g gm 80 mu chim t l 47,9%, tip sau l cnh g 53 mu chim t l 31,8%; sn phm chim t l thp nht l i g ch c 2 mu chim t l 1,2%.

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Biu 4.2: S mu tht g ng lnh nhp khu c xt nghim trong thng 3/2012.

4.2.

Kt qu pht hin Salmonella trn mu tht g nhp khu Tng cng 167 mu kim tra ch pht hin c 2 mu c khun lc in hnh. T

2 mu xt nghim nghi ng dng tnh vi Salmonella, chng ti kim tra v c kt qu nh sau: Bng 4.4: T l nhim Salmonella trn cc loi sn phm tht g ng lnh nhp
Loi sn phm cnh g i g i g 1/4 M i g Chn g G nguyn con i ti g Tng Cng S lng 53 2 80 4 12 3 13 167 s mu nhim 0 0 2 0 0 0 0 2 T l 0% 0% 2.50 % 0% 0% 0% 0% 1.20 %

Da vo bng 4.2 ta thy c 2 mu nhim Salmonella u thuc loi sn phm i g 1/4, chim t l 2,5%.

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Hnh 4.2. Khun lc Salmonella c trng trn mi trng HE v XLD

tin tng sinh

Tng sinh trong RVS

Cy chuyn sang XLD v HE

Hnh 4.3. Mt s bc tin hnh phn lp Salmonella spp.

Hnh 4.4. Khun lc Salmonella trn mi trng TSA

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a).

b). Hnh 4.5. Kt qu th sinh ha xc nh Salmonella spp. a).Kt qu sinh ha Salmonella dng tnh b).Kt qu sinh ha Salmonella m tnh

4.3.

Phn tch t l nhim vi khun Salmonella trong sn phm tht g ng lnh nhp khu

Qua kt qu thc t cho thy trong s cc mu c xt nghim th ta c 2 mu cho kt qu l dng tnh vi Salmonella trong tng s 167 mu, chim t l 1,2%. Biu 4.3: T l nhim Salmonella trn tng s mu xt nghim.

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C 2 mu nhim u phn b vo loi sn phm i g, nguyn nhn m loi sn phm ny c t l nhim cao hn cc loi sn phm khc c th l do: Nh chng ta phn tch trn, Salmonella l mt loi vi khun c mt khp mi ni trong t, nc... Nu nh trong qu trnh chn nui, khng c bin php phng nga trit th y c th l ngun gy nhim vi khun trong thc phm sau ny. C th l do vt nui n phi thc n b nhim Salmonella ri t nhim vo i l phn sau cng ca con g gn vi hu mn, l ni c th tip xc trc tip

thc phm. vi phn v cc cht thi b. Nu nh g n phi thc n b nhim vi khun Salmonella, vi khun ny theo ng phn gy ly nghim vo i. Trong qu trnhgit m, khng m bo v sinh nc bn c mang vi khun nhim vo thc phm hoc do tay ngi git m khng m bo v sinh tip xc vo thc phm. Ngoi ra, cn c mt s nguyn nhn khc c th gy nhim Salmonella vo thc phm nh l: tp nhim t cc dng c, vt cha khng m bo hoc tp nhim t qu trnh vn chuyn.

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PHN V: KT LUN V NGH

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5.1.

Kt lun Qua kt qu xt nghim 167 mu tht G ng lnh nhp khu, chng ta nhn

thy: trong 167 mu tht G ng lnh nhp khu bao gm cc sn phm: cnh g, i g, i ti g, i g , chn g, g nguyn con khi xt nghim ch tiu salmonella th c 2 mu cho kt qu dng tnh vi Salmonella, chim t l 1,2%. C 2 mu ny u thuc loi sn phm l i g 1/4, cho thy quy trnh ch bin cha t yu cu, do cc l hng tng t cng cn c kim tra chc ch hn. T kt qu kim tra ch tiu Salmonellani ring v kim tra vi sinh ni chung, cho php chng ta ra bin php x l trit cc sn phm thc phm nhp khu trc khi c vo th trng nhm m bo an ton tuyt i cho ngi tiu dng. Tt c cc mu khc u t tiu chun quy nh (chim 98.8%), iu ny cho thy rng cc sn phm tht g ng lnh nhp khu vo th trng nc ta qua kim tra vi sinh l tng i sch, phn nh iu kin t khu nguyn liu, ch bin, ng gi n bo qun l t yu cu, m bo an ton cho ngi s dng. 5.2. ngh Hin nay vn An Ton V Sinh Thc Phm ang l vn ton cu v cng ngy cng xut hin nhiu chng vi sinh vt gy bnh nguy him hn cho con ngi. Cho nn vic tm hiu kin thc chuyn ngnh tng qut t kin thc trong sch v trn ngh nh trng v kin thc t thc tin s gip sinh vin chng em nm vng hn v kin thc chuyn mn. Do , nh trng nn to iu kin, lin kt vi cc trung tm phn tch, kim nghim t gi sinh vin ca mnh n hc hi nhng kin thc thc tin v bin nhng kin thc hc c trn sch v thnh nhng cng vic c ch, v cn nhm gip sinh vin c iu kin tri nghim, nng cao kin thc ca mnh t nhng kin thc thc t, v cao hn na l c th ng gp mt phn cng sc ca mnh vo lnh vc nghin cu trong tng lai; Bn cnh l gip nh trng hon thin chng trnh Hc i i Vi Hnh.
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TI LIU THAM KHO 1. V Minh Chu-C quan th y vng VI- Ti liu hng dn cng vic hng ngy 2. 3. 4. 5. . .1,2)

6. L Th M-Phn lp v gim nh mt s vi khun hiu kh thng gp ng tiu ha ca d khe mnh v d mc hi chng tiu chy nui ti cc tnh Nam Trung B- Lun vn Thc s Nng nghip. 2006. 7. 8. 9. 10. . 11. Nguyn Nh Thnh v cng s-Vi sinh vt th y, 1997.. 12. g . . . .

13. L Ngc T-c t hc v an ton thc phm 14. 15. . 16. Ti liu Cc phn ng sinh ha, Trung tm k thut tiu chun o lng cht lng 3 (Quatest3) 17. ISO 6579:2002, FDA Chaper 5-2005 18. WHO Global Salmonella Surveillance (2000-2004)
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19. W.Blackburn, Peter J.Malure.Foodborn pathogens Hazards, rick analysis and control 20. Eugene W.Nester, Denise G.Anderson, C.Evans Roberts Jr, Nancy

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44 SVTH: PHM NGC H - 09077661