Bacterial Population in Intestines of the Black Tiger Shrimp (Penaeus monodon) under Different Growth Stages.docVIP

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Bacterial Population in Intestines of the Black Tiger Shrimp (Penaeus monodon) under Different Growth Stages.doc

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Bacterial Population in Intestines of the Black Tiger Shrimp (Penaeus monodon) under Different Growth Stages

Bacterial Population in Intestines of the Black Tiger Shrimp (Penaeus monodon) under Different Growth Stages WaniladaRungrassamee1,AmornpanKlanchui1,SageChaiyapechara2,SawarotMaibunkaew1, SithichokeTangphatsornruang3,PikulJiravanichpaisal2,NitsaraKaroonuthaisiri1* 1Microarray Laboratory, National Center for Genetic Engineering and Biotechnology, Khlong Luang, Pathum Thani, Thailand, 2Aquatic Molecular Genetics and BiotechnologyLaboratory,NationalCenterforGeneticEngineeringandBiotechnology,KhlongLuang,PathumThani,Thailand,3GenomicResearchLaboratory,National CenterforGeneticEngineeringandBiotechnology,KhlongLuang,PathumThani,Thailand Abstract Intestinalbacterialcommunitiesinaquaculturehavebeendrawntoattentionduetopotentialbenefittotheirhosts.To identifycoreintestinalbacteriaintheblacktigershrimp(Penaeusmonodon),bacterialpopulationsofdisease-freeshrimp werecharacterizedfromintestinesoffourdevelopmentalstages(15-day-oldpostlarvae(PL15),1-(J1),2-(J2),and3-month- old(J3)juveniles)usingpyrosequencing,real-timePCRanddenaturinggradientgelelectrophoresis(DGGE)approaches.A totalof25,121pyrosequencingreads(readinglength=442624bases)wereobtained,whichwerecategorizedbybarcode for PL15 (7,045 sequences), J1 (3,055 sequences), J2 (13,130 sequences) and J3 (1,890 sequences). Bacteria in the phyla Bacteroides,FirmicutesandProteobacteriawerefoundinintestinesatallfourgrowthstages.Therewere88,14,27,and20 bacterialgeneraassociatedwiththeintestinaltractofPL15,J1,J2andJ3,respectively.Pyrosequencinganalysisrevealed thatProteobacteria(classGammaproteobacteria)wasadominantbacteriagroupwitharelativeabundanceof89%forPL15 and 99% for J1, J2 and J3. Real-time PCR assay also confirmed that Gammaproteobacteria had the highest relative abundanceinintestinesfromallgrowthstages.Intestinalbacterialcommunitiesfromthethreejuvenilestagesweremore similartoeachotherthanthatofthePLshrimpbasedonPCAanalysesofpyrosequencingresultsandtheirDGGEprofiles. Thisstudyprovidesdescriptivebacterialcommunitie