VACCINES S DNG VI KHUN SNG NHNG B C CH C LC TRONG NTTS

Tm tt
Chng nga l mt chin lc qun l quan trng c s dng duy tr sc khe con ngi v ng vt trn ton th gii.Pht trin vc-xin trong nui trng thy sn lm gim vic s dng khng sinh trong sn phm dnh cho c. Vc xin truyn thng s

dng cho c c ngun gc t vi khun ( git cht vi khun bng formalin) c a vo c th c thng qua vic ngm hoc tim to ra dch th (khng th) min dch. Vc-xin th h tip theo da trn khng nguyn bt hot cng vi mt cht b tr nng cao hiu qu vc xin. sn xut tloi khng nguyn gy bnh b lm suy yu, khng cn kh nng gy bnh nhng vn c tc dng gy phn ng min dch bo v cho c th.
Trong nhng nm 1990, bt u pht trin chin lc ng dng vc-xin sng trong cc loi c. MLV l mt dng vaccines s dng vi khun cn sng c loi b tc nhn gy bnh hoc loi b c lc bng phng php ha hc, vt l, hoc cc phng php k thut di truyn. MLV thng vn gi c kh nng ly nhim sang cc vt ch nhng MLV s dng kh nng ca cc khng nguyn l bo v bng cch to ra min dch t bo (CD4, CD8 T-t bo phn ng). Mlv thun li ch chng c th d dng a vo c th c (v d, bng cch ngm c con) v kch thch c hai kh nng min dch dch th v t bo trong thi gian di . Bt li bao gm cc vn an ton vc-xin i vi vt ch v mi trng. Thnh cng ca MLV c p dng trong nui trng thy sn mi trng nc m c s dng lm ni bt chin lc vc xin sng . Tim chng l chin lc hiu qu c s dng ton cu nhm kim sot bnh truyn nhim cc loi ng vt. Vaccines c bit ln u l vaccine

phng bnh u ma cho ngi bng cch s dng vi - rt sng lm vaccines. Tuy nhin, nhc im ca vaccine bt hot l i hi phi sn xut mt lng ln t bo, ng thi, d c hiu qu trong vic kch thch sinh khng th nhng hiu qu to p ng min dch qua trung gian t bo khng cao. Trong khi vaccine nhc c, s dng vi khun sng b lm yu c lc lm vaccine v khng

cn thm cht b tr, li rt c hiu qu trong vic kch thch p ng min dch qua trung gian t bo cng nh duy tr tr nh min dch v khng cn phi to mt lng ln t bo trong vic sn xut vaccine.
Nm 1961, theo Jenner vc-xin bo v con ngi khi vi - rt bnh u ma bng con ng t vi - rt nga, bu v b. Tizard (1999) cung cp mt xem xt tuyt vi trong ngnh vc-xin i vi con ngi v ng vt th y. Tht th v, phn nhiu trong s vaccines sm s dng c Modified Live Vaccines da trn s thnh cng ca vaccines bnh u ma. S tng cng nui c dng lm thc n v cho chng trnh tch tr gia tng s hin din ca bnh. Bnh l biu hin do nhiu yu t, bao gm cc iu kin mi trng khng c quan tm to iu kin d dng cho s ly truyn ca mm bnh trong c nui ti mt tch tr cao. Duff(1942) l ngi u tin bo co trn th nghim vaccines trong c hi m Bc M, Oncorhynchus clarki, bng cch cho n vaccines git(vaccines bt hot) Aeromonas salmonicida. nui trng thu sn, vc - xin hot ng sm tp trung vo vic s dng killed vc - xin. Vc - xin c cp php v mt thng mi u tin cho c l killed vc - xin c thc hin bng phng php ngm( dng ngm) chng li Yersiniaruckeri tc nhn gy bnh xut huyt di hu. Sau s thnh cng ca sn phm ny,vaccines dng ngm s dng formalin git cht vi khun c p dng cho c hi phng bnh dovibriosis c pht trin. Nguyn l tng t cho s kh hot tnh ca mm bnh vi khun c hi Atlantic, Salmo salar, c s dng pht trin vaccines h c hi v p dng phng php ngm. Phng php ngm vaccine chng li A. salmonicida khng hiu qu v vy vaccine vi khun bng phng php chch ln u tin c thc hin(vaccine s dng theo phng php tim ra i). Bricknell v O'Dowd nghin cu phn ng min dch ca cc (vaccines bt hot) A salmonicidia khc nhau git vaccine trong c hi Atlantic khng s dng cht h tr(b tr). Vaccine polsaccharide ngoi t bo(ngoi bo) (EPS) gy ra phn ng khng th v c bo v trong khong 2 thng sau khi chch(tim). O'Dowd s dng vaccine c cha formol git vi khun(vaccines bt hot bng formalin) to ra t lp in dng hoc lp in m A.salmonicidia pht trin iu kin hn ch st gy min dch cho c hi Atlantic. Kt qu ca vic ny dn n phn ng khng th ng k c nng cao sau khi chng nga liu cao ; tuy nhin, c khng thch ng c. Bricknell s dng s kt hp gia EPS v lp in m vaccine vi khun to c khng th v bo v

c 8 tun sau khi tim chng nga ( 90% so phn trm sinh tn). Xy dng vaccines tng t ny mang li hiu qu bo v ( 60% so vi phn trm sinh tn ) ln ti 9 thng. Hu ht mi vaccine hin nay c dng trong ngnh cng nghip sn xut c hi Atlantic da vo khng nguyn ( vi khun v virus ) vi du l cht h(b) tr, cung cp trong mt liu chch(v c a vo c th c bng phng php tim) ( Sommerset et al.2005 ). Vaccines ny( c ng dng thnh cng) thnh cng v gim s dng ho cht bt li i vi mi trng, c bit l khng sinh, nh sn xut c hi thng mi c gia tng ( Markestad v Grave 1997 ). Gi tr sn phm c nng cao cho php s dng chin lc tim chng trong cng nghip c hi. Bi ph bnh mi y v s dng vaccine trong lnh vc nui trng thu sn Chile cho thy phng php ngm(Bi bo co mi y cho thy vic ng dng vaccines trong cng nghip nui trng thy sn Chile s dng c 2 phng php ngm v tim). (Vic tim chng nga cho c phi tim t lc cn nh)chin lc tim chng c phi chng t lc cn nh ( 10-12 ngy sau khi th ging) hoc kch thc nh ( 1-2 g c hi non ) l khng thit thc. Thm d v s dng l bt u vo nhng nm 1990 (Norqvist et al.1989; Vaughan 1993; Thornton 1994; Lawrence 1997; Hernanz moral 1998; Mars-den 1998; Klesius and Shoemaker 1999; Thune 1999). Hin nay, ba loi Vaccines c ch c lc vi khun c quyn s dng Hoa K, bao gm vc - xin chng li bnh thn vi khun(Renogen1), nhim trng huyt rut ca c nheo M (AQUAVAC-ESC ) v bnh Columnaris (AQUAVACCOL ). (Bi bo co ny tp trung vo) chin lc gim c lc cho Vaccines c ch c lc vi khun, u im v nhc im ca s dng Vaccines c ch c lc vi khun trong nui trng thu sn v hin ti, v d thnh cng ca Vaccines c ch c lc vi khun thng mi thng dng trong nui trng thu sn mi trng nc m Attenuation Strategies Laboratory Passage Nhiu phng php c s dng lm suy gim tc nhn gy bnh cho s pht trin thnh cng ca modied live vaccines ngi v trong y hc th y. Cc phng php c s dng cho n nay trn cc tc nhn gy bnh ca c s c nu r. Mt trong nhng k thut u tin l simple laboratory passage thng qua cc sinh vt trn cc mi trng hoc nui cy m dn n s suy gim ca cc vi sinh vt gy bnh. Mt bng sng ch u tin m t cng dng ca k thut ny vi cc c nheo M, Ictalurus punctatus, virus (CCV) (Hart-man v Noga 1980). H to ra mt CCV gim c lc bng cch thng qua cc virus trong nui cy m bng cch s dng mt dng t bo c ngun gc t walking catsh, Clarius batrachus. Gn y hn, Daly et al.(2001) s dng laboratory lm suy yu chng Renibacterium salmoninarum chng nga cho c hi i Ty Dng. H phn lp hai chng ln m chng d dng pht trin trn

mi trng nui vi khun thng thng v chng minh c s bo v sau khi tim vaccine. Itano et al.(2006) cng cho thy vic s dng mt chng phn lp c c lc thp Nocardia seriolae nh l mt chng vc-xin tim nng v chng minh c s bo v khi nhng thch thc nguy him.Tuy nhin, s phn lp ny khng lm suy yu hon ton trong c trc ui vng(yellowtail), Seriola quinqueradiata, ng vt ch. Vi khun mi trng Mt k thut th hai lin quan n vic thng qua cc tc nhn c lc trong mt vt ch thay th (tc l, khng nhy cm vi tc nhn gy bnh) hoc s dng mt loi vi khun mi trng m phng cu trc khng tc nhn gy bnh. ng dng thnh cng nht ca chin lc ny trong nui trng thy sn l s dng Arthrobacter davidanieli c cho tim vo bo v c hi t vi khun gy bnh thn gy ra bi R. salmoninarum (Griffiths et al 1998; Salonius et al.2005). Cc tc gi cho thy mt mi quan h pht sinh loi gn gi gia A. davidanieli v R. salmoninarum v kh nng gy phn ng cho gia huyt thanh min dch vi carbohydrate b mt (Griffiths et al. 1998). Gn y hn, Itano et al.(2006) th s dng mt chin lc tng t (tc l, Quan h pht sinh loi v phn ng khng nguyn cho) xc nh vaccines thch hp chng li N. seriolae. Trong nghin cu ny, cc tc gi s dng mi trng ca loi Nocardia v nh gi kh nng ca cc phn lp (N. soliton v N. fluminea) to ra min dch bo v chng li N. seriolae khi tim vo Yellowtail. Tht khng may,s phn lp ny ch cung cp mt s bo v gii hn thch thc vi c lc ca N. seriolae. t bin ha hc v vt l Chin lc th ba lin quan n t bin ha hc hoc vt l m kt qu l s t bin ngu nhin (s) trong tc nhn gy bnh, vi chng vc-xin c la chn b c ch c lc trong ng vt ch bi vic thiu tnh c hi trong ng vt ch (Linde et al 1990; Klesius v Shoemaker nm 1999; Shoemaker et al. 2007). V d tt nht l vic s dng rifampicin, mt loi khng sinh cho kt qu lm suy gim vi khun gram m bng cch cm ng thay i trong lipopolysaccharide (LPS), mt yu t c lc quan trng (Schurig et al 1991; Klesius v Shoemaker 1999; Arias et al.nm 2003; Zhang et al.2006). Thng tin chi tit v nhng thay i trn LPS gy ra bi on rifampicin c tho lun chi tit trong cc v d c cung cp trong bi bo ny. Chin lc ny l mt trong nhng vic mang tnh tch cc nht cho n nay qua cc thnh tu v vc-xin th y (Linde et al 1990; Schurig et al 1991; Kle-sius v Shoemaker 1999; Gantois et al 2006;. Shoemaker et al 2007.). K thut di truyn Chin lc th t l k thut di truyn in hnh bng cch chn vo, xa, hoc gin on con ng chuyn ha (s) hoc gen c lc (s) m kt qu lm suy gim tc nhn gy bnh. Cooper et al.(1996) s dng mt on gen nhy mini (mini-transposon) lm gin on vic sn xut ca chondroitin sulfatase (yu t nghi ng c tnh c lc) trong E. ictaluri. C nheo M c t bin cho thy s bo v khi nhng thch thc nguy him vi yu t c lc c phn lp t chng b m hoang di. Mt n lc

gn y l phng php to t bin bng gen nhy lm mt tc nhn gy c O polysaccharide khi phn lp E. ictaluri v c s dng nh mt loi modied live vaccine (Lawrenceet al 2001;. Lawrence v Banes nm 2005). Cc tc gi thnh

cng trong vic phn lp cc dng E. Ictaluri thiu Opolysaccharide nhng tht bi trong vic chng minh s bo v ca vic phn lp ny i vi vic ngm chng nga c da trn (Lawrence and Banes nm 2005).
Igarashi v Iida (2002), bng cch s dng mt cng ngh tng t, bo co v s pht trin ca mt vaccine gim c lc E. tarda bng cch to ra mt chng t bin E. tarda (t bin gen nhy) vi vic sn xut siderophore thp hn bo v c r phi, Oreochromis niloticus, chng li nhng thch thc gy cht. Leung et al. (1997) cng to ra mini-Tn5 (t bin gen nhy) gy ra s tng trng v thiu protease A. hydrophila cho vic s dng nh modied live vaccines trong blue gourami (Trichogaster trichopterus). Random transposon (Tn917) mutagenesis and subsequent screening trong c vc lai c sc (Morone chrysops X M. saxatilis) to ra mt Streptococcus iniae vi mt gen phosphoglucomutase b t qung (Buchanan et al 2005). Enzyme

phosphoglucomutase c cho l quan trng cho s hnh thnh v bao polysaccharide trong vi khun.S c mt hay vng mt ca mt v bao c tha nhn l quan trng i vi tnh c (Barnes et al 2003;. Buchanan et al 2005). S phn lp c tc dng lm suy gim tnh c
trong c vc lai c sc v cho thy s bo v khi nhng thch thc gy cht nu modified live S. iniae c chuyn vo c bng cch tim vo phc mc (bng) (Buchanan et al 2005.). Mt trong nhng chin lc ph bin hn l s dng cc chng t bin auxotrophic (Hoiseth v Stocker 1981, Smith et al 1984; Vaughan et al 1993; Lawrence et al 1997; Hernanz Moral et al 1998; Thune et al 1999; Temprano etal 2005). Thng thng, gen aroA ca tc nhn gy bnh c bt hot bng cch chn mt DNA cassette c cha mt gen nh du khng khng sinh la chn khi trao i allelic bng cch s dng mt vector bt hot(suicide). S ngng hot ng ca gen ny khng cho php tn ti trong cc vt ch v s cn thit ca hp cht chuyn ha cha nhn thm. Tuy nhin, liu cao ca cc t bin ny c th gy t vong ng vt ch (Lawrence et al 1997;. Thune et al 1999). Mc d chin lc ny to ra vaccine gim c lc nhng thng s phn lp gim c lc ch c th bo tn trong thi gian ngn (24-72 gi) v do khng kch thch min dch y trong cc loi c con. u v nhc im ca vc xin bin i trc tip: An ton:

Mt trong nhng nhc im chnh khi s dng Modified live vaccines chnh l an ton. C kin cho rng cc loi khng khng sinh (
killed vaccines) hoc sn phm khng sinh th an ton khi s dng i vi hu ht cc loi ng vt di nc bi v tc nhn gy bnh b tiu dit hoc b bt hot( do tc dng ha hc hoc do x l nhit). Vn ca Vc xin git(vaccines bt hot) chnh l khng c kh nng khng vc xin (c kh nng pht tn mm bnh). Mt trong nhng vn khc ca vc xin git chnh l kh nng kt dnh sau khi tim vo c hi khng nguyn ca vc xin vi b tr. i khi, kt qu l l gim s tng trng ca nhng loi ng vt c tim nga vc xin v lm mt i cht lng ca sn phm nh s sn chc ca phn tht c.Mt trong nhng quan im v s an ton khi s dng Modified live vaccines chnh l chng to ra vc xin khng c kh nng gy bnh cho ng vt c tim nga vc xin. Hn

th na ,vn v mi trng cng ang tr thnh 1 im ang ch , v kh nng pht tn ca chng sn xut vc xin c th ly lan trong mi trng. Frey (2007) a ra mt nh gi
tuyt vi v khi nim an ton sinh hc v mt di truyn ca Modified live vaccines khi ng dng trong sn xut thuc cha bnh cho ng vt. l nguyn tc c bn p dng trong vic s dng Modified live vaccines c ng dng trong ngnh nui trng thy sn.

M ,theo b nng nghip Hoa K(USDA), c quan kim sot ng thc vt Hoa K(APHIS) cc chuyn gia ca cc th y thy sn cho php ng k v cp giy php .Cc nghin cu v mc an ton c tin hnh bao gm cc th nghim liu lc tim nga gp 10 ln v c tim trc tip t c qua c. V theo nh phn tch mi nguy th cn cn phi kho st n kh nng pht tn ca vc xin trong mi trng.cng vi s hin din ca mm bnh trong mi trng t nhin v kh nng mm bnh ly nhim qua ngi cng ang l 1 vn cn c cn nhc k lng.
Bo v min dch: Mt trong nhng u im ca Modified live vaccines chnh l n c kh nng lm kch thch hot ng ca t bo trung gian,th dch ( khng th) v nim mc min dch (Clack v Cassidy Hanley 2005). Modified live vaccines n tn ti v c sao chp cng vi vt ch, chnh l kt qu ca s phn ng mnh ca t bo min dch nhm bo v n trong thi gian di. S Cm ng tnh min dch t bo (CD4+ v CD8+ t bo phn ng T ) c nhim v to mng bo v chng li s nhim trng ni bo ((Seder v Hill 2000). Cc nghin cu gn y chng minh c rng c mi lin h trc tip no

trong phc h tng hp m (Major Histocompatibility Complex _MHC) gia lp I (Antao et al. 2001) v lp II(Goodwin v cng s nm 2000) trong c. S hin din ca khng nguyn vi chnh xc ca MHC cho php cc phn ng v nhn bit c thng qua cc du hin nhn bit tng ng t t bo phn ng T v B. Seder v Hill (2000) cho thy rng Modified live vaccines c th to ra c Th1 v CD8 t bo phn ng T. Marsden et al. (1996) l ngi u tin chng mnh c kh nng ca vic s dng Modified live , n c th loi b vc xin th t bin A. salmonicida v u tin tng cng t bo T phn ng trn t bo B trong vic s dng Modified live vaccines trong c hi sc Oncorhynchus mykiss. Hin nay dc trn phn tch v sn phm lin quan n phin m ca cc gene biu hin trn gan c tr cho thy c cch sng ca VK,gene MHC lp I cng vi acute phase response genes c tng cng hiu chnh cho thy rng cc khng nguyn ch ng x l v th hin (Peatman et al. 2008). S cm ng gene MHC lp I ch ra rng CD8 t bo phn ng T b nhim vi khun ni bo.phn ng kch thch theo sau mt nhm T bo phn ng T chnh l c kh nng sn xut ra cht c ch s sinh trng ca Virus (interferon gamma) (Milev-Milovanovicet al. 2006) v qu trnh sn sut c th lm vt trung gian ph hy tc nhn gy bnh ni bo (Seder and Hill 2000). V vy da trn gi thuyt ta c th to nhng phn ng min dch t nhin da vo vic ng dng Modified live vaccines cho c. Seder v Hill (2000) tip tc ch ra rng vaccine git hoc nhm vaccine phn t nh khng c kh nng kch thch phn ng t bo. Vc-xin git, hoc cc khng nguyn git, kt qu s cm ng ca dch dch th (khng th trung gian) min dch. Vaccine git Edwardsiella ictaluri trc y c s dng trong nui trng thy sn c chng minh khng xm nhim vo c (Nusbaum v Morrison 1996) v do , khng th to ra s min dch. Trong mt s trng hp, qu trnh git hi (v d, iu tr formalin) c chng minh l thay i khng nguyn b mt quan trng (Bader v cng s 1997). C 2 nhn t c coi l hng u trong c kh nng kh hot tnh ca vc xin lm chng bt hot v khng th bo v c c con trc s nh hng bt li ca intracellular pathagens ( Thune et al 1997). Hu ht cc vc xin git c pht tn sau khi tim vi s h tr ca cht b tr. Ban u vibrio bacterins c pht tn ch yu khi c ngm di nc, mc d hiu qu bo v tt nht c biu hin sau khi tim(Norqvist et al 1989). Thi gian min dch gy ra bi cc loi vc-xin b git l thng t hn 4 thng v ch c hiu qu trn cc mm bnh hoc tc nhn gy bnh tit ngoi c t. M rng di ca ph min dch ca vc-xin b git thng da trn chng nga v/hoc cc cht b tr. Sau khi min dch tip xc vi trc tip vi khun (t bo min dch trung gian) c chng minh c tc dng hn 4 thng trong c tr l chng nga duy nht.(klesius) Vn chuyn vc xin : Modified live vaccines c u im chnh l d dng phn phi trong ng vt c tim chng thng qua ng tiu ha khi cho n hoc ung hoc khi ngm c trong vc cin.(Norqvist). Modified live vaccines pht trin trong ngnh cng nghip thy sn nhng vng nc m hoc phn phi bi phng thc ngm exposure to the youngest life stage ( prior to release into ponds). Modified live vaccines ngn chn kh nng xm

chim v ly nhim cho vt ch iu cho thy hiu qu ca vic pht trin phn phi khi ngm vc xin. Modified live Vibrio anguillarum vaccines c chng minh ch vi mt lng nh vc-xin ngm sau khi tim nga li to ra c mc bo v cao (Norqvist et al 1989). Norqvist et al. (1989) ngh Modified live vaccines c xu hng nhn rng trong vt ch v do lm tng tn hiu vc-xin. Gi c: Chi ph vc xin l mt cu hi ln i vi quan h sn xut v s hng ng ca nng dn. Modified live bacterial vaccines i vi hu ht cc phn l chi ph thp trung bnh (Seder v Hill 2000; Klesius et al 2004) . Vi khun gim c lc c th c s dng sn xut nhiu lt vc-xin quy m ln men thng mi. Cc chi ph ch yu l tinh khit, an ton, xc nh immunodose, tim nng (hiu qu) th nghim, v ng gi (v d, s ng kh ) . For formalin killed vaccines, th gi c tng t nh killed Y. ruckeri vaccines c s dng ph bin cho c hi. Mc d, iu cn thit phi x l l timchng vo tng c th trong ngnh cng nghip c hi i Ty Dng cn tiu tn 1 lng chi ph lao ng ng k. mt trong nhng chin lc mi c a ra chnh l p dng cng ngh ti t hp DNA v cng ngh tim chng DNA v chng cng i hi chi ph tn km hn v cn phi tinh sch khng nguyn ti t hp v chi ph u t cho vic phn lp v tim kim c tnh ca gene(s) mong mun p ng s mim dch khi tim chng vo vt ch. cng ngh tim chng DNA c s dng thnh cng i Ty Dng, bo v chng li virus ly nhim hoi t mu (IHNV) (Simard et al 2006) v c hi sc bo v chng li virus xut huyt nhim trng huyt (VHSV) (Lorenzen et al 1999). Vic chp nhn ng dng vc-xin DNA mt s nc (v d,v an ton thc phm v cc vn sinh vt bin i gen) c th l mt yu t hn ch trong vic thc hin thng mi ca cc sn phm ny (Babiuk et al. 2000). Example Nn tng Nhim trng huyt rut ca c nheo M ( ESC ) gy ra do E. ictaluri, vi khun Gram m ban u c m t vo nm 1976. Hng nm,chi ph mt mt v bnh ESC trong nn cng nghip c tr Hoa K ln n 40-60 triu $.Vic pht trin formol git vi khun E. ictaluri trong vaccine c thc hin, nhng chng khng c hiu qu bo v chng li ESC. Nhc im ca sn phm l khng to min dch ch ng cho c sau khi tip cn bng phng php ngm. Phng php tin tng kim sot ESC l cho n vi ch n ung c cha thuc khng sinh ; tuy nhin, cch thc hin ny l khng hiu qu v c gim vic hp th thc n v theo di cho thy vi khun c kh nng khng khng sinh. Min dch bo v

Nghin cu trn E. ictaluri gi rng khng th l quan trng cho phn ng min dch bo v. Klesius v Sealey thy rng qua dn truyn th ng, khng th c trng khng mang tnh cht bo v. Antonio v Hedrick thy rng t bo khng th c hiu s c tc ng bng cch cung cp thuc chng vim, kt qu gia tng tnh mn cm ca c nheo gp 2 ln khi tip xc vi E. ictaluri. S v K l ngi u tin chng minh min dch t bo chu trch nhim v bo h. C c dng trong nghin cu ca h c gy min dch bng cch cho tip xc vi hm lng thp ca c t E. ictaluri. Nh vo chng nga, i thc bo c thu hoch t c cn sng st v nh gi cho kh nng git E. ictaluri trong ng nghim. i thc bo t c c chng nga E. ictaluri c th git E. ictaluri ( 85% ) ; trong khi, i thc bo t khng chng nga hoc c gy min dch vi vc-xin cht th hiu qu km hn( 68.1 v 71.4% ). Hn na, c sng st liu thp tip xc vi E. ictaluri c khng v thch ng tip theo vi ng hnh v i khi phn lp d hnh. Nghin cu trn cho thy nhu cu cn c vaccine s dn n kt qu t bo min dch trung gian c phn ng min dch trong c tr con S pht trin ca E. ictaluri c thay i:

Klesius v Shoemaker (1999) pht trin mt loi vc-xin c ch c lc ca vi khun E. ictaluri thng qua vic phn lp c lc trn mi trng BHI agar (brain heart infusion agar) c b sung rifampicin. K thut ny c s dng cho cc vi khun Gram m to ra t
bin th (Schurig et al.1991) v c s dng nh vc-xin. Kiu hnh th c c trng t vic thay i trong LPS ca vic phn lp cc chng b m. S thay i ny lm mt mt chui O side ca LPS nh thy trong vaccine RB-51 Brucella abortus. Nghin cu gn y m t t bin ca E. ictaluri RE-33 chng minh s thay i l trong LPS. t bin c cho thy s thiu trng lng phn t cao ca cc LPS khi so snh vi vic phn lp chng b m (EILO).

(Hnh1) bi immunoblot (Klesius v Shoemaker Nm 1999; Arias et al. 2003). Kt qu ca s thay i ny lm suy yu kh nng xm nhp v s bn b ca E. ictaluri c da trn to cc phn ng min dch thch hp nhng khng gy bnh (Klesius v Shoemaker 1999. Mt s cc kim tra b sung c tin hnh phn lp xc nh cc loi vc xin khc nhau (RE-33) bng cc th nghim sinh ha hoc axit bo. Cc thng tin ban u thu c t vic phn lp cc chng b m v cc vaccine t bin l khng c s khc bit v cc thng s sinh ha, ngoi tr vic vaccine phn lp pht trin trn mi trng c b sung rifampicin (Klesius and Shoemaker 1999). Tuy nhin, bng cch s dng h thng nhn dng vi khun (Microbial ID, Inc., Newark, DE, USA) th s khc bit trong cu hnh ca acid bo c xc nh. Nhn chung, s khc bit v s lng trong cu hnh ca acid bo vi vic phn lp th ch c mt vi acid bo c mt trong sc ph t vic phn lp cc chng b m EILO (3-hydroxy hexadecanoid acid) hoc phn lp t bin RE-33 (hepatdecanoic acid, 3-

hydroxy-heptadecanoic acid, v 10-methyloctodecanoid acid), cho php xc nh chnh xc. (Arias et al. 2003). S khc bit trong th nghim sinh ha c pht hin vi Biologs (Biolog,Hayward, CA, USA) s dng h thng carbon xc nh vi khun. H thng biolog chng minh s tng ng vi t l 73% (Pearson product moment correlation v do , cho php c s khc bit gia cc chng vaccin v cc chng b m c phn lp. (Arias et al. 2003) An ton. Vaccin phn lp c chng minh l rt an ton thng qua con ng trc tip t c sang c v cho thy khng c kh nng nhim virut tr li. Cc kim tra an ton c thc hin mi ln v cho thy khng c phn ng bt li no sau khi tim. (Klesius and Shoemaker 1999). Th nghim c thc hin t 10 30 ging c tra c nh nc v cc bc s th y Mississippi v Alabama ph duyt nm 1997 v khng c bo co tc dng ph no ca vic tim phng (Klesius and Shoemaker 1999). Bao gm cc th nghim an ton c thc hin bi Intervet Intervet, Inc. (1998 and1999) hn 57 triu c da trn v cc ging c c tim nga p ng cc yu cu cho mt sn phm an ton. Khng c vn no c bo co c c tim chng t cc ni m th nghim c tin hnh. Hiu qu. Hiu qu c chng minh trn 3 - 9 ging c tra khng E. ictaluri c mt trong vaccin c ch c lc ca vi khun ny (Klesius v Shoemaker nm 1999; Lawrence Banes 2005). Liu vc xin c bin i trong phm vi t 105 107 n v to thnh khun lc (CFUs) RE-33/mL v c tip xc vi c bng cch ngm t 2 30 pht. Th nghim c chng minh l c hiu qu min dch i vi c nng 107 CFU/mL c iu kin th nghim ln c thch thc bn ngoi E. ictaluri. Th nghim cng chng minh rng (2000) hiu qu vaccin lin quan n s khc bit di truyn ca cc ging c trong h c da trn. Su ging c trong h c da trn c th nghim v RPS c tm thy nm trong khong t 67,1 n 100% vi mt kt qu c ngha trn c h l khong 80%. Nhng kt qu ny cho thy dng c da trn c th pht trin v

p ng tt hn sau khi tim chng. Mt ngun tin khc t hiu qu ca s bin i trong cc nghin cu l s phn lp E. ictaluri c s dng trong th nghim chng nga c da trn (Klesius and Shoe-maker 1997). Khng nguyn bin th tn ti trong E.ictaluri lin quan n s min dch tp nhim. Chng ti tm thy s bo v khng c tin cy da vo tt c cc th nghim. S bo v c nhn thy da vo vic phn lp t 8 13 chng vi RE-33 E. ictaluri (Klesius and Shoemaker 1999). Hiu qu cng c xc nh bng cch s dng cc rifampicin khc to ra E.ictaluri t bin (B-21909; B-21910; B-21911;Agricultural Research Service Culture Collection, National Center for Agricultural Utilization Research, Peoria, IL, USA). Bng 1 cho thy s phn lp cc loi vaccine khc nhau c th cung cp s bo v chng li nhng thch thc ng hnh v d hnh (Klesius and Shoemaker 2000). Mt th nghim vaccine a gi c s dng vi s kt hp ca rifampicin v cc chng t bin b suy yu ca E.ictaluri s (ATCC 202058 = RE-33; B21909; B-21910; B-21911) cc t l khc nhau mang li mt liu vaccine tng s 107 CFU/ml (Table 2) (Klesius and Shoemaker 2000) cng c tin hnh. Kt qu cc th nghim a khng chng minh rng mt vaccine da trn t bin suy yu rifampicin ca E. ictaluri l kh thi v hiu qu. iu ny l ht sc quan trng nu hin ti ch phn lp c mt loi vaccine duy nht tr nn km hiu qu. S thnh lp cng thc ny c th c thay i hoc sa i bng cch thm mt hoc mt vi loi vaccine c phn lp cung cp mt s bo v vng chc nu mt loi khng nguyn khc tr nn chim u th trong ngnh cng nghip c da trn. Hiu qu ca vaccine trong dng c da trn non (712d post-hatch) v trng c c chng minh (Shoemaker et al. 1999; Wise and Terhune 2001; Klesius et al. 2002; Shoemakeret al. 2002; Shoemaker et al. 2007). Petrie-Hanson and Ainsworth (1999) cho rng c da trn khng th hnh thnh h min dch (khng th trung gian) cho n 4 tun sau khi n. Hanson v Ainsworth (2001) chng minh s hin din ca t bo min dch trong m to mu thn v l lch 3 ngy sau khi n. Warr (1997) pht hin chc nng ca t bo lympho trong cc c quan lymphoid c da trn mt vi ngy sau khi n. Hu ht cc hot ng tha nhn rng cc t bo min dch trung gian chu trch nhim v vic bo v

chng li E. Ictaluri (Shoemaker v Klesius nm 1997; Shoemaker et al.1997). S pht trin kh nng min dch thay i dng vaccine l hu nh c kh nng bi v s hin din ca cc i thc bo hoc khng nguyn hin din trong t bo c con hoc s tn ti ca chng vc-xin cho n khi h thng min dch c p ng. iu ny c chng minh sau y trong in ovo administration of live viral vaccines in poultry (Mast v Goddeeris 1999). Theo nh hiu bit, c c th cho kt qu thnh cng trn nhiu loi vaccine 710 d u trng mi la trng n hoc trng c (tim 24 72h) trc khi n l quan trng nu c s thay i ca cc dng vaccine khc (virus hoc vi khun) tr nn c gi tr v c s dng trong nui trng thy sn. Bin php chin lc ny cho php vic chng nga sm nht c th c (tc l, trong tri ging) trc khi a vo mi trng sn xut.

S thay i c ch c lc ca E. ictaluri RE-33 c cp bng sng ch (US Patent no. 6,019,981)v c cp giy php ca Intervet, Inc. by the USDA-Agricultural Research Service. Intervet, Inc. (Millsboro, DE,USA) th trng vaccine thay i vng i ESC (2001 n nay) di nhn hiu AQUAVACESC as an USDA-APHIS-CVB. Nm 2006, sn phm thay i thnh dng ng kh vi cng thc ng lnh. Hiu qu c ti nh gi v chng minh vi cc sn phm ng lnh t nht 65 ngy sau khi tim chng (Wise, 2006). Th nghim cho thy s dng cc kt qu vc xin. Th nghim ngh s dng kt qu ca vc-xin ging c ln hn, ci thin chuyn ha thc n, v ci thin mt s tr li ca $ 3900 - $ 4900 cho mi ha c tim phng so vi c khng tim phng (Intervet Inc 2003; Wise, 2006).

Carriaset al. (2008) gn y bo co v vic s dng vaccine c ch c lc thm vo cng tc m rng sn xut ging / giai on vn m. Kt qu cho thy mt s ci thin tr li (tin bc v s sng cn) c chng nga cho c c t chc trong mt vn m trc khi a vo cc ao c ging (Carriaset al. Nm 2008). Theo bo co v ngnh cng nghip c da trn ca M cho thy trn 25% ging hoc ging sn xut mi nm c chng nga vi AQUAVAC-ESC. Cc dng c da trn sn xut c th v ang s dng vc-xin ny trong k hoch qun l sc khe hiu qu (ESC).

Kt lun
Vaccines c ch c lc vi khun c s dng thnh cng trn ngi, th y, v ng vt sng nc ngn nga bnh.Nu nguy c tim n n ng vt, mi trng v con ngi u thp hoc khng ng k th s dng Vaccines c ch c lc vi khun trong nui trng thu sn l chin lc thch hp . Nh chc trch cn xem xt tim nng vaccine trong mi trng khi khng c nhiu d liu khoa hc dn chng v c lc ca vi khun cn tim tng. Vaccines l cng c c dng bo v sc khe vng chc v an ton sinh hc dn n li ch ln cho nh sn xut nui trng thu sn