Title. Bharadwaj, Manish Surendra. Citation ( ) Issue Date Right

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1 NAOSITE: Nagasaki University's Ac Title Author(s) The transferability of drug resista Photobacterium damsela subsp.piscic Bharadwaj, Manish Surendra Citation ( ) Issue Date URL Right This document is downloaded

2 Åëlaeegedeff6eGeemeeeecgeEdifisceessfi*wa r Sa,KmandTcwereineffectivefoffP.desmselassp. pisciciaa,becaaxseofhighirmcidenceofffesistantstraimesawad hightransferabilityoftheirffesistapmces.fewunexplaimaable reasens,kmresistancehasspreadamemegp.aesmselassp. piscicidastrainsiftjapaneventhoughkmhasneverbeen usedasanafttibacterialagentagainstinfectionsinfish farms.cpifesistantp.daneselassp.piscicidastrairs decreasedconsiderabgyaftertheyeara989,thereasonfor whichcamebeexplainedbythefactthattheuseofcpagainst bacterialinfectionsoffishfarmswasifestrictedinyapaft since1983.thisstudyhasreportedforthefirsttixrkeinp. damselassp.ptsciciaanoteniythatsofirestraimsharboured resistancetoem,butalsothattheemffesistancewas traptsferrable.also,thisstudyhasreportedforthefirst timeinrdaneselassp.ptsciciaathatmxcfortmpexhibited trixkxedaidistributienviz.sensitiveimtermediate-eeve1s resistantandhigh-levegresistant.thisdifferenceixx resistaflcelevelsoftmpmeaybeeitherdueterrtaere differencesinthegroupofstraiansusedinthisstudyerit maybeduetodifferentffesistapmcemechameismsfer intermediate-levelapmdhigh-ieveltmpresistances,eke 112

3 tffansferabilityefdrugresistapmcesoccasredonxywhenthey exhibitedresissancestotc,natc,ornatrrftp."rchereasopmfex thisabilityeftheseresistancesfteedtobestudied thoroanghly. Difugifesistancesaretffansfefffedwithtwodifferent freq"encieswhenapresistanceistransferred,with resistancesincludingaptraitsferringatafrequencyof abomut1om6amedresistancesincludimgsatrapmsferffingata frequencyofabout10m4.furtherinvestigationshowedthat inthedormersthattffansferredapresistaitce,dtfug resistancesineachdonorwerenormaelytraftsferredby twoplasmidsviz.asmaee6skbplasmidencodingdrug resistancesincludingapffesistance$ap,cp,emand/ortc resistances$andagaxger12skbplasmidcodingfor resistapmcesinciudingsa(km,saand/ortcresistances). therefore,transcorjugantsharbouringboththe6sand 12Skbplasmidsusuallycarriedalithetransferab]e resistancesincludingapandsaresistances.simcethe transferabilityofapresistamecevariedwithrkhedffug resistaxtcepatteritirthedopmors,ithiittsasthepessibigityof theapresistancebeingtrasusferredbyatransposition mechanismsixkxiaartothetn3faitkily. 113

4 VpoitcgoRingandsequencingtheP-lactamasegene thatconferffedapresistanceinp.damselassp.piscicida $PDP-AP$,itwasfoupmdteshowRittlehoxRftolegywithother krownp-gactamaases.rhededucedagxitinoacidsequencesof PDP-APshowedmaximumsimilaritywiththeclassAPlactamases,Moreover,thefouxixxxportaRtstructwrai featuresfoundcoptservedimcgassap-lactamaseswere presentinthededucedamimoacidsequenceofpdp-ap. Hence,itbeloitgstoceassA.As,expected,thehydropathy plotofthefi-lactamasesequencedshowedittobeasoluble pretein.webelievethatthe6-aactamasegenehasbeen transferredbyatn3liketransposonontoanoriginaiiy crypticplasmidofp.aamselassp.pisciciaaintowhichother ffesistanceshadalreadyaccumulated.tethebestofour knowledge,thisisthefirsteverreportofa6-racsamase genebeingisglatedandsequencedfromafishpathogemeic bactefia,andproposesanaltogethernewcoftceptforap resistaneeipmp.aamselassp.ptscieiaa. Yeiiowtaillsfedwithvarioassdrugsasfeedadditivesdopmot causedrwagresistanceofbacteriaintheimtestina]twact.also, theintestinaltractofyelgowtailldoesnotfavourthe traitsferofdrugresistawacegenesbekweenbacteriaof differentspecies.so,ithasbeeftpresumedthasthetransfer 114

5 ofresistancebesweeitintestinalbacteriaandp.d between intestinal bacteria and damsela esmsela ssp. ssp, piscicida piseiciaadeesnottakepaaceiymtheintestineofyellowtaig does take place in the intestine of yellowtail. e 115

6 Claeepeeecfijr.vaRwaeeaecaswaÅëkwask*mps emheoccmeifrenceandtffansferabieityofdffeng ffesesgancewirkh respectto14antirk\)icrobiaacempoasftdsagainsk 183sgrains ofp.daneselassp.piscieidaisolatedfrom culturedyemaowtail Seriolageeinqeradiatawasipmvestigatedand theconelusions are asfoglows:- a. Ofthesotaldrugresistancesi.e.Ap,Cp,E]rm,Km,Na,Sa,Tc andtmp,alltheresistancesexceps coajugarlytransferffedteescherichiacol Rpr.lhetransferofdrugffesistancewas thepatternandievelofdrugresistances thedonoifstrains. Nacouldbe ik-12x1037 depemedeftton harboredby 2. Regardingthestrainsthatdidnotcarry ertransferap resistance,arethedrugffesistanceswere transferrredon asingie12skbpgasmidwithatxansfer frequencyof 1O-4e 3e Regardimgthestrainsthattffawasferred Apresistance, thedifugresisgapmeeswerecarifiedby twoplasxrftidsviz.a a2skbanda6skbplasmid.thetraftsfer frequencyof thefor]nerwas10b4aptdthatofthelaterwas 1Oun6. 4'e Withrespecttothestrainsthat transferffedap 116

7 resistarce,resistancestokm,saand/oroccassionailytc werecarriedonthe12skbpiasrr\)id,andresistancesto Ap,Cp,EmandoccassionaRlyTcwerecarriedonthe6S Kbplasntaidinthetransconjugants. Se Apresistance(fi-lactamase)genethatwasisolatedhad anorfof846bpsofdnathatcodedfor282aminoacids,andhadacalculatedmolecularmassof31,292da. 6e "rcheamino-acidsequenceofthep-lactamaseshoweda maximumhomologyof56%.ingeneral,theaminoacid sequencehomologywasmuchhigherwithgramnegativebacteriaascomparedtogram-positivebaeteria. Also,uponcomparisonwiththevariousclassesofPlactarnases,thehighesthomology(S69o)wasseenfor ClassA6-lactamases,Inaddition,ahydropathyplotof thep-lactamaserevealedittobeagiobularsoluble enzyme. 7. Viablecountsofbacteriaintheintestinedecreased remafkablywhenyellowtailswerecultivatedwithfeed additivesinthepreseneeofvariousdrugs. 8e Bacterialfioraintheintestineofyellowtaiiconsisted maineyofcoryneformbacteria,staphylococcessand Bacillblsregardlessofwhetherdrugswereadministered ornot.gram-positivebacteriawereabundartinthe 1fi7

8 intestine.also,significawatdiffereftcesinthe couldnetbeobservedfortheixresistamece microfloras etorespecggve dffugsaxitkongtheisoratesfromefisheulltivated withfeeds cometainingoffptotcormtainingeachdraxg. ffence,no tifansferofdrengresistancetakesplaceinthe tractofyeleowtaiisbetweentheintestinalbacteriaandp. damselassp.piscicida. ifitestina1 M8

9 RewERENÅëES A]eeabile-mCasevas,C.F.&Chiourell,M.E.(X992).Bacterial prasmidsandgepmefesux.cell70,a89-fl99. AxEkb1er,R.P.(198O).ffhestrwactuffeofS-lacSamaases.Philos TransRSocLonaBBiolSci a.s Amabler,R.P.,Coulsoft,A.F.W.,Frere,j.M.,Ghuysen,Joris, B,,Foffsman,M.,kevesque,R.C.,Tiraby,G.&Waley,S.G. $199a$.AstandardnumberingschemefortheCkassAPiactaxnases.Biochemf ' Armdersome,E.S.(1968).theecoRogyeftransfewabRedrug ffesistanceiwatheenterobacteria.annrevmicrobiol22p 131ua180e Akashi,A.&Aoki,T.(1986).Charactexisationof transferablerplasxxxidsfrorrkaeromonashyarophila. NipponSutsanGakkaishiS2,649rem6SS. Aoki,T.(1974).Studiesofdrug-resistantbacteriaisolated fromwaterofcarp-pordsandirtestinaltractsefcarp, NipponSutsanGeskkaishi40,247-2S4. Aoki,T.(197S).TransferofRfactorintheintestinagtracts effishandsterilizedwaterofcu1turedpopmd.fishpathol9, 16ew-a73. Aoki,T.(k992).Chemotherapyanddrugifesistanceinfish 119

10 faffmsinsapan.in:shariff,m.,subasinghe,r.p.&arther,g. R.(eds),DiseasesimeAsiaftaqenacaxIteeifeK,AsianFisheries Society,Manila,Phiglipines,Si9-S29. Aeki,T.,Axai,T.&Egusa,S.(a97ew).betectionofRplasmids innatwffaeryoccwaringfish-pathegermicbacteria,edsiyarasiella taraa.mierobiollmneesnol2a77-83.s Aoki,T.,Egusa,S,&Arai,T.(1974).DetectiofiofRfactorsin RaturaliyoocuringU/ibrioangeeillarmemstrains. Antimicrobialagentsandchemotherapy6,S34-S38. Aoki,T,,Egusa,S.,Ogata,Y.&Watanabe,T.(197a),Detection efresistancefactorsinfishpathogenaeroneonas ligecefesciens,fgenmicrobtol6s, Aoki,T.,Egusa,S.&WataRabe,T.(1973).betectionofR" bacteffiaimcueturedmarinefish,yeigowtaix$seriola qutnqeseraaiata$.fpnjmicrobiola7,7-12. Aoki,T.,Egusa,S,,Yada,C.&Watanabe,T.(1972).Studiesef drugifesistanceandrfactorsinbacteriafrompondcurturedsaamoptids.g.amago$dncorhymechessrhodesress macrostoneess)andyagnaxxxe(0becorhynchecsmasao ishikawae$.fapesnfmierobiola6, Aoki,T.&Kitao,T.(a98a).Difugffesistanceandeffaftsferable RpgasmidsinEdwarasiellataraafremfishou1tureponds. mo

11 FishPathol1S277-28a.s Aeki,T.&Kitao,T.(a98S).glbetectienoftffaymsferableR prasxtitidsimstraifisofthefish-gpathogexticbacteriuitg, Pessteeerellapiscicides.fFishDis8,34S-3SO. AekisTepKitaO,TepIeMewapNesMitO]rmapYe&NOMUwasTe(1983)e ThesassceptibilityofAeromonassalmonicidastrainsisolated incukufedandwildsalmonidstovarious chemetherapeutics.nipponsutsangakkaishi49, Aoki,T.,Kitao,T.&Kawapmo,K.(1981).Changesindrug resistanceofvibrioangesillareeminculturedayu, PlecoglossusaltivelisTemaminckaxtdSchlegel,ingapan.fFish DiS4233mp"230e' Aeki,T.,Kitao,T.,Watanabe,S.&Takeshita,S.(1984).Drug resistaitceandrplasmidsinvibrioangesillareeneisoiatedin cas1turedayu(plecoglossessntttvelis).microbiolimmesnol28, 1ma9e AokR,T.,Sakaguchi,T.&]Kitao,T.(1987).MultipgedrugifesistantplasmidsfromeEdwarasiellamrdesineelcukure ponds.nipponsutsangakkaishis3,a s. Aoki.,T.,Takami,K.&Kitao,T.(i990).Dfirugresistanceina non-hemolyticstveptococcesssp.isolatedfromcugtured yegrowtailsertolagutnqeraaiata.disaqeeat0rg8,a7x-g77. Aoki,T.&Wataitabe,T.(1973a).SeudEesofdrugresistaitce ve1

12 andrfactorsinbacteriafrompond-cukuffedsrlmaemeids.eg. Rainbowtroant(Salmogairareerif.iriaeess).ewshPnthol8, 83pm89e Aoki,T.&Watanabe,T.(1973b).Studiesofdrasg-resistant bactexeaisolatedfromeel-pondwaeerarmdintestinagtifacts oftheee1(angeeillajoponicaandangecillaaregecilla).nippon SutsanGeskkatshi O.s Bishop,R.&Sherratt,D.(1984),TwannsposonTn:1 intramoleculartfanspositiofi.molgengenet196,a ElwelE,L.P.(1994).RplasrEgidsandantibioticresistances.in: MillerVL,KaperJBZB,PortnoyDA,XsbergRR(eds).Molecugar GeneticsofBacteria1Pathogenesis.AmericaRsocietyfer MicrobioRogy,Washingteit,DC.,pp.a7-41. Gauthier,G.,hafay,B.,Ruimy,R.,Breitt]neayer,V.,NicoEas,Y. L.,Gauthier,M.&ChristeR,R.(199S).Sxkitala-subunitrRNA sequeneesandwholednarelatednessconcurforthe ffeassignmentofpasteecrellapiscicides(sniszkoetal.)janssen andsuffgallatothegenusphotobaeterinmas Photobactertumaaneselessubsp.ptseicidacomb.nov.fntJ SystBaetertol4S,139-a44. GroSasJeHe,TOIftkaSkY,MeEepACtiSpLeAesFalkOWpSe(a994)e Plasmids,InMethoasforgeneralandmoleeutar bactertology,pp.36s-386.editedbygerhardt,p.,murway, 122

13 Ce: ReGeEesWOOdsWeAe&Kfirie&NeReWaShimgtOnsDe AmericapmSocietyfexMierobioAegy, Hanahan,D.(g983).Studiesontransfermatiepm of Escheriehiaeoliwithplasmaids.fMolBiolfi66,SS7-S8O. Hedges,R.W.&jacob,A.F.(g974).TffansposEtiopm ampicilginresistancefromrp4teotherreplicon.mol ef Gen Gereet s HeffrOnsFepMCCarthYpBejesOhtSUbO,He&OhtSUbgsEe(1979) e DNAsequenceanaEysisofthetransposonTn3:three genes andthfeesitesi"volvedintranspositioftoftn3.cell 18p 11.S3op1.IE.64e Heffron,F.,Subtett,R.,Hedges,R.W.,gacob,A.&Falkow, $197S$,OifiginoftheTEMbeta-lactamasegeitefound Se on prasmids.fbacteriola22,2so-2s6. HOEtpSeGe,SneathpPe]E{eAepMairsNeSe&SharPeMeEe(1986)e BergeryNsMananalofSystematicBacteriology,Vo]. 2e WiRliaxxks&Wilkins,Baltimore. Iyobe,S.,Sagai,H.&Mitsuhashi,S.(g981).Tn2001,a eransposonencodingchleramaphenicogffesistamece in Pseesdonvonasaereeginoses..7Bescta46,a4a-A48. gyobe,s.,tsumeoda,m.&mitsuhashi,s.(a994).cloning and expressioninenterobacteriaceaeeftheexgerded-spectrum 6-aactamaasegefiefromaaPseecdoneonessaeresginosesplasgrmid. M3

14 FEMSMierobiolLettk21a7S-a8O.s a61e. JapaneseSocietyofChemotherapy(a98a).Therevised standardmethodferdeterminingmiceminimuminhibitory Jaitsserm,W.A.,&M.j.SasffgaiAa(1968).Merphology, physigfogy,andserogegyefapastenvellaspeciespathogewtc ferwhiteperch(roccasamericareees).fbacteriol96,16o6- cowacentwation].chemotherapy29,76-79(enjapanese). Kado,C.I.&Liu,S.T.(1981).RapidprocedurefordetectioR andisolationoflargeandsmallplasmids.fbact14s,a36s- 1373e Kawahaxa,E.,]Kawai,K.&Kusuda,R.(1989).Invasionof Pasteesrellapisciciaaintissuesofexperimentailyinfected yellowtailseriolaqeciregeresainta.nipponstsisangakkaishi SiSs499va"SO1-e ]Kim,E.&Aoki,T.(1993a).Drugresistanceandbroad geographicaxdistributionofidenticalrprasmidsof Pessteesrellapiscicidesisoiatedfromacugtuffedyegkowtailipm Yapan.Microbtollmmeereol37,1O3-1O9. KiXtskpEeHe&AOki,T.(1993b).lhestructureofthe chleramphepticokresissancegeneoitatffarmsferabrer peasxe]idfremthefishpathogen,pessteesrellapiscieides, Microbtollnemesnol377OS-7i2.p 124

15 ]sttao,t.&aoki,t.(1976).mierobia]ffoffaefarteficiaediets. FishPashol1O181-a8S.p Kusuda,R.,llkaoka,M.&Kawai,K(g988).Dffasgsensitgvityof PasteesrellapiscicidastraiftsisolatedfromcasXtuffed yel]owtai]froma1984tea98s.nipponsutsangakkaishis4, gs2a--as26. Kusuda,R,&Kawai,K.(a998).BacterialdiseasesofcuEtured rrgarinefishingapapm.ptishpathol33,22a-227. Kusuda,R.,Sugiura,S.&Kawai,K.(199O).]Drugsensitivity ofpastemerellapiscicidastrainsisolatedfromculltuifed yeggowtaiifroitm1986toa988.nipponsutsangeskkaishis6, 1:!l39E"24:i2e Kusuda,R.&Sugiyama,A.(198a).Studiesonthe charactersofstaphylococonsepiaermiaisisoiatedfroma diseasedfishes.i:\(inthemorphologicag,biologicarand biochemicarpropersies.fishpathol16,1s-24. Kusuda,R.&Yamaoka,M.(1972).Etiolegicalstudieson bacteriaipseudotubereugosisimacuaturedyelliowtaigwith Pessteesrellapiscicidaasthecausativeageftt.a:Oifathe gwaorphologtca1andbiochemaicagprropereies.nipponsutsesme Geskkaishi38a32S-1332.p Kyte,3.&]boggittle,R.F.(g982),AsiliEkplexxxethedfoff displayingthehydrepathiccharacterofaproteim.fmolbtol 125

16 IS7aOS-132.p hederberg,e.m.&coher,n.(1974).tifansforxeitasionof Satneonellatyphimecrtumbypgasnkiddeoxyffibepmwacieicacid. fbesctaagao72-ao74.p hewissidehlepgrumblesslecesmccormel,se&feowerspaelle $a970$.pasteurella-likebacteriafffomeaepizooticift menhadenandntaulaetingalvestonbay.foesrnalofwildlife aiseases6a6e-162.s Matagne,A.&Frere,j.-M.(199S).Comtributiofiofmutant analysistotheunderstandingofenzymecatalysis:thecase efclassap-lactamases.bioehimicaerbtophysieaactaa246, 1O9-12'":7'. Milger,J.H,(1972).ExcperimentsinMolecalarGeitetics,Cold SpringHarborLaboratoryPress:ColdSpffingHarborr,NY. Miller,R.V.&Kokjohn,T.A.(1990).GeneraAmicrobiologyof reca:enviropmmentaaandevolutionafysignificance.annes RevMicrobtol4436S-394.s Minami,T.(1979).Streptococcesssp.,pkthogeflictecultured yelzowtaill,isolatedfroxnfishesfewdiets.ewshpatholk4,as- 19e Merii,H.,Captxt,D.C.,Taylor,L.Y.&Murray,C.K.(a986). Ferxmaatiowaofhistagg\)EnebykumiitousbacteriaisoAatedfro]caR scembroidfish.nippovesutsangakkatshis2,213s-2fi

17 Morii,ff.&Kasarrxga,K.(1989).Baceerialflgrainthedigestive tractsofmaandskepperfishesboleophthalmesspeetinirostrts andpertophthalmesseantonemesis.nipponsutsesngeskkutshi iss'ai]733-'i]74[g-.p Muffoga,K.,SugEyarrtrxa,T.&UekiN.(1977).PasteuregXosisEn cukuffedbgackseabream(myliomacrocephains).foeermealof thefeseecltyoffishertesaredameinealffeesbareary,ffiroshimes Untversity,16,17-2a. Murpky,E.(a989).TxaftsposabgeegementsinGrampositivebacteria.in:Berg,D.E.&Howe,M.M.(eds).Mobile DNA.AmericanSecietyfortheMicrobielogy,Washingtopm,D. Cesi;269tu288e Nielsepm,H.,Engellbrecht,J.,Bmeanak,S.&Heipae,G.V.(a997). Edentificatiopmofprokaffyoticapmdeukaffyoticsignaipeptides andpredictieneftheixcaeavagesites.proteinengao,a-6. Sa]mabroek,J.,Fffitsch,E.F,&Mapmiatis,'ff.(a989).Motecutax Cloiting:ALaboratoryManual,2madedft.ColdSpfEymgHarbor Labewatoffy:ColldSpringHarbew,NY. Sehaberg,D.R.,Highsmith,A.K.&WaChSmaUthsIeKe(1977)e ResistamecepgasmaidtransferbySerratianearceseeresin ewemee.antineicrobagentschemaotherla,449-4se. SnieSZkOpSeFesBUllkOCksGe]LesHOIkiSpEe&Beeymes3eGe(A964)e Pasteeerellasp.fromaRepizgoticofWhatePerch\(Roccas 127

18 esmeerieesness\)imchesapeakebaysidewageraffeas.fbaeeertol 88g814-18gS.p Stokes,H.W.&HaM,R.M.(a989).AnovellfamEllyof petepmeiagayrrxaobilednaeleffxaentsepmcodipmgsitc-specefic gefte-imetegffatiopmfunctions:integrofis.molmicrobtola2, a669-s683. Sera,H.&Ishida,Y.(a972).Bacterialfloffainthedigestive tractsofgeftarinefish.mfill:cgassificatigpmofisogatedbactexia. NipponSutsanGakkaishi38,8S3-8S8. Sherffatt,D.(a989).Tme3andrelatedtvanspos.abReeAemeents: Site-specificffecembinatiopmaitdtffanspositEon.MmeMobile DNA,pp.a63-a84.EditedbyE.B.DougRas&M.H.Martha. Washingten,D.C.:AmericarmSocgetyforMicrebioXogy. ShiKxkidu,U.(1986).Marimebacteria.in:NaitouA(ed). ResearchanaUtilixuttonofMarineBtologicalResoecrees.CMC, Tokyo.69-82(ipmSapaxxese). Shimidu,U.&Egusa,S.(a972).Affe-examinationofghe fish-pathogenicbacteriumathathadbeepmffepertedasa Pasgeecrellaspecies.NipponSutsanGakkaishi.38,8O3"m8a2. Sugtyapmka,A.&Kusuda,R.(a98Xa).Studiesonthe ÅíkaracSersofStaphylococcasepiderneiatsisogatedfroma diseasedfiskes.ig:sererogica1pifepertiesoftheisolates.f7ish fpnthola62s-33.p $28

19 Seegiyamaa,A.&Kusuda,R.(1981b).Studiesepmghe characsersefstaphylococcasepiaermidgsisogatedfroma diseasedfishes.fiig:acomaparasivestasdyoftheserekogicar pxepertiesbetweentheiseaatesandthestffainsofhumame eregin.fishiipatholg6,3s--4a. Takashima,N.,Aoki,T.&Kitao,T.(198S).Epidemiogogicafl suffveillapmceefdffa]g-resistafitstragnsofpasteecrella ptscicida.fishpathol2o,2o9-2a7. Takemura,I.&Kassuda,R.(k988).EffectofJosarrmeyctu adigxinistfatieftontwtestinagmxxicrofloraofcukuffed yeaaewtair.ni,ppponsutsengakkaishis4,837-84o. firamigaschr,m.(g982).experiftltentenperoraginocuiatienvia foodtoinduceyeakowtaiastrepsococcicosis.nipponsecisan Geskkaishi4817a7-a72O.p kopsewepessanwantopae&pohficele(2000)enovelppm RactarrftKakasegenesfrorriSwoenvironmentalisolatesof Vibrtoharveyi.AntimicrobAgentsCheneother44(S), fl3og-a3g4. Teranzo,A.E.,Barja,J.L.,Colwell,R.R.&HetrickF.M,(g983). ChewacterizagionefpkasxnidsRnbacteriagfishpathegens. Infecttonandlmmaesnity39,g84-a92. Watameabe,T.,Aeki,T.,OgaSa,Y.&EgusaS.(a97fl).Rfactors ffellagedsefrshculsuripmg./zrenny/icesdscia82,383-4ao, 129

20 Watson,J.D.,Hopkins,N.H.,Reberts,J.W,SgeitzJ.A.& Weiner,A.M.(g987).MolecuRarbioXegygfthegepte.TRke Bemejarrxxin/CumffkRtwgsPublishingCompamey,Enc.,MenkePark eec Yasupmaga,N.,Hatai,K.&Tsukahara,J.(a983).Pasteesrella piscicidafxomanepizooticofculturedxedseabream.fish Pathologya8,1O7-aaO. Zhao,Y.&Aoki,T.(1992).Plasmidpxofiaeaflalysisof PastexrellaptsciciaaanduseofapgasmidDNApffobeto identifythespecies.faqmeaaninenthealth4,g98-2o2. ZhaOpJesKEMsEesKObaYaShisTe&AOki$Te(1992)elkUg resistanceefvibrtoangutllaruueisoaatedfromayubetween a989and1991.nipponsutsangakkatshis8,1s23-as