Analysis of the large bowel microbiota of colitic mice

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1、LettersinAppliedMicrobiology2005,41,45–51doi:10.1111/j.1472-765X.2005.01720.xAnalysisofthelargebowelmicrobiotaofcoliticmiceusingPCR/DGGE11221R.Bibiloni,M.A.Simon,C.Albright,B.SartorandG.W.Tannock12DepartmentofMicrobiologyandImmunology,UniversityofOtago,Duned

2、in,NewZealand,andDepartmentofMedicine,UniversityofNorthCarolina,ChapelHill,NC,USA2004/1276:received6November2004,revised21January2005andaccepted25February2005ABSTRACTR.BIBILONI,M.A.SIMON,C.ALBRIGHT,B.SARTORANDG.W.TANNOCK.2005.Aim:Totestcombinedpolymerasechai

3、nreactionamplificationof16SrRNAgenesequencesanddenaturinggradientgelelectrophoresis(PCR/DGGE)asananalyticalmethodtoinvestigatethecompositionofthelargebowelmicrobiotaofmiceduringthedevelopmentofcolitis.MethodsandResults:Thecolonicmicrobiotaofformerlygermfreein

4、terleukin10(IL-10)-deficientmicethathadbeenexposedtothefaecalmicrobiotaofspecificpathogen-freeanimalswasscreenedusingPCR/DGGE.ThecompositionofthelargebowelmicrobiotaofIL-10-deficientmicechangedascolitisprogressed.DNAfragmentsoriginatingfromfourbacterialpopulati

5、ons(‘Bacteroidessp.’,Bifidobacteriumanimalis,Clostridiumcocleatum,enterococci)weremoreapparentinPCR/DGGEprofilesofcoliticmicerelativetonon-coliticanimals,whereastwopopulationswerelessapparent(Eubacteriumventriosum,Acidophilusgrouplactobacilli).SpecificDNA:RNAdo

6、tblotanalysisshowedthatbifidobacterialribosomalRNA(rRNA)abundanceincreasedascolitisdeveloped.Conclusions:PCR/DGGEwasshowntobeaneffectivemethodtodemonstratechangesinthecompositionofthelargebowelmicrobiotaofmiceinrelationtoprogressionofinflammatorydisease.Theint

7、ensityofstainingofDNAfragmentsinDGGEprofilesreflectedincreasedabundanceofbifidobacterialrRNAinthemicrobiotaofcoliticanimals.AsbifidobacterialfragmentsinPCR/DGGEprofilesgeneratedfrommicrobiotaDNAshowedincreasedintensityoffragmentstaining,anincreaseinbifidobacterial

8、numbersincoliticmicewasindicated.SignificanceandImpactoftheStudy:PCR/DGGEanalysisdemonstratedanalteredcompositionofthelargebowelmicrobiotaofcoliticmice.Thisworkwillallowspecificgroupsofbacteriatob