Microbial Diversity of Bovine Mastitic Milk as Described by Pyrosequencing of Metagenomic 16s rDNA 英文参考文献.docVIP

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Microbial Diversity of Bovine Mastitic Milk as Described by Pyrosequencing of Metagenomic 16s rDNA 英文参考文献.doc

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Microbial Diversity of Bovine Mastitic Milk as Described by Pyrosequencing of Metagenomic 16s rDNA 英文参考文献

MicrobialDiversityofBovineMastiticMilkasDescribed byPyrosequencingofMetagenomic16srDNA GeorgiosOikonomou,ViniciusSilvaMachado,CarlosSantisteban,YnteHeinSchukken,Rodrigo CarvalhoBicalho* DepartmentofPopulationMedicineandDiagnosticSciences,CollegeofVeterinaryMedicine,CornellUniversity,Ithaca,NewYork,UnitedStatesofAmerica Abstract Dairycowmastitisisanimportantdiseaseinthedairyindustry.Differentmicrobialspecieshavebeenidentifiedascausative agentsinmastitis,andaretraditionallydiagnosedbybacterialculture.Theobjectiveofthisstudywastousemetagenomic pyrosequencingofbacterial16SrRNAgenestoinvestigatebacterialDNAdiversityinmilksamplesofmastiticandhealthy dairy cows and compare the results with those obtained by classical bacterial culture. One hundred and thirty-six milk samples were collected from cows showing signs of mastitis and used for microbiological culture. Additionally, 20 milk sampleswerecollectedfromhealthyquarters.BacterialDNAwasisolatedfromthesamemilksamplesandthe16SrRNA geneswereindividuallyamplifiedandpyrosequenced.Discriminantanalysisshowedthatthegroupsofsamplesthatwere most clearly different from the rest and thus easily discriminated were the normal milk samples from healthy cows and thosecharacterisedbycultureasTrueperellapyogenesandStreptococcusspp.Themastitispathogensidentifiedbyculture were generally among the most frequent organisms detected by pyrosequencing, and in some cases (Escherichia coli, Klebsiellaspp.andStreptococcusuberismastitis)thesinglemostprevalentmicroorganism.Trueperellapyogenessequences werethesecondmostprevalentsequencesinmastitiscasesdiagnosedasTrueperella pyogenesbyculture,Streptococcus dysgalactiae sequences were the second most prevalent sequences in mastitis cases diagnosed as Streptococcus dysgalactiaebyculture,andStaphyloccocusaureussequenceswerethethirdmostprevalentinmastitiscasesdiagnosedas Staphylococcusaureusbyculture.Insamplesthatwereaerobicculturenegative,pyrosequencingidentifiedDNAofbacteria that are known to c