NAOSITE: Nagasaki University's Ac Title Author(s) The transferability of drug resista Photobacterium damsela subsp.piscic Bharadwaj, Manish Surendra Citation (2002-03-31) Issue Date 2002-03-31 URL http://hdl.handle.net/10069/7218 Right This document is downloaded http://naosite.lb.nagasaki-u.ac.jp
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
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
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
ofresistancebesweeitintestinalbacteriaandp.d between intestinal bacteria and damsela esmsela ssp. ssp, piscicida piseiciaadeesnottakepaaceiymtheintestineofyellowtaig does take place in the intestine of yellowtail. e 115
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
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
intestine.also,significawatdiffereftcesinthe couldnetbeobservedfortheixresistamece microfloras etorespecggve dffugsaxitkongtheisoratesfromefisheulltivated withfeeds cometainingoffptotcormtainingeachdraxg. ffence,no tifansferofdrengresistancetakesplaceinthe tractofyeleowtaiisbetweentheintestinalbacteriaandp. damselassp.piscicida. ifitestina1 M8
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