Culture-independent identification of gutbacteria correlated with the onset ofdiabetes in a rat model肠道文化独立的鉴定 细菌与发病 糖尿病大鼠模型

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1、TheISMEJournal(2009)3,536–548&2009InternationalSocietyforMicrobialEcologyAllrightsreserved1751-7362/09$32.00www.nature.com/ismejORIGINALARTICLECulture-independentidentificationofgutbacteriacorrelatedwiththeonsetofdiabetesinaratmodel11211LuizFWRoesch,Graciel

2、aLLorca,GeorgeCasella,AdrianaGiongo,AndresNaranjo,13143AriannaMPionzio,NanLi,VolkerMai,CliveHWasserfall,DesmondSchatz,431MarkAAtkinson,JosefNeuandEricWTriplett1DepartmentofMicrobiologyandCellScience,InstituteofFoodandAgriculturalSciences,Universityof2Florid

3、a,Gainesville,FL,USA;DepartmentofStatistics,UniversityofFlorida,Gainesville,FL,USA;34DepartmentofPediatrics,UniversityofFlorida,Gainesville,FL,USAandDepartmentofPathology,ImmunologyandLaboratoryMedicine,UniversityofFlorida,Gainesville,FL,USABacteriaassociat

4、edwiththeonsetoftype1diabetesinaratmodelsystemwereidentified.Intwoexperiments,stoolsampleswerecollectedatthreetimepointsafterbirthfrombio-breedingdiabetes-prone(BB-DP)andbio-breedingdiabetes-resistant(BB-DR)rats.DNAwasisolatedfromthesesamplesandthe16SrRNAge

5、newasamplifiedusinguniversalprimersets.Inthefirstexperiment,bandsspecifictoBB-DPandBB-DRgenotypeswereidentifiedbyautomatedribosomalintergenicspaceranalysisatthetimeofdiabetesonsetinBB-DP.LactobacillusandBacteroidesstrainswereidentifiedintheBB-DR-andBB-DP-sp

6、ecificbands,respectively.SangersequencingshowedthattheBB-DPandBB-DRbacterialcommunitiesdifferedsignificantlybuttoofewreadswereavailabletoidentifysignificantdifferencesatthegenusorspecieslevels.Asecondexperimentconfirmedtheseresultsusinghigherthroughputpyros

7、equencingandquantitativePCRof16SrRNAwithmoreratspergenotype.Anaverageof4541and338116SrRNAbacterialreadswereobtainedfromeachofthe10BB-DRand10BB-DPsamplescollectedattimeofdiabetesonset.NinegeneraweremoreabundantinBB-DPwhereasanotherninegeneraweremoreabundanti

8、nBB-DR.ThirteenandelevenspeciesweremoreabundantinBB-DPandBB-DR,respectively.Anaverageof23%and10%ofallreadscouldbeclassifiedatthegenusandspecieslevels,respectively.QuantitativePCRverifiedthehigherabunda