SI

S1SupportinginformationforBacterialcommunitystructureinthedrinkingwatermicrobiomeisgovernedbyfiltrationprocesses.AmeetJ.Pinto1,ChuanwuXi2,LutgardeRaskin1*1DepartmentofCivilandEnvironmentalEngineering,UniversityofMichigan.2DepartmentofEnvironmentalHealthSciences,UniversityofMichigan.*Phonenumber:[phone].Emailaddress:[e-mail]:10pageswithfivefiguresandthreetablesareincludedbelow.S2FigureS1.SchematicoftheAnnArborDrinkingWaterTreatmentPlant.Opendiamondsmarkthesamplinglocations.Thedualmediafiltersarebackwashedwithfinishedwaterandthebackwashwaterisintroducedintotheequalizationbasinattheheadoftheplant.Thedistributionsystemwassampledat13differentmonitoringpoints.S3FigureS2.AmountofDNAextractedonamassbasis(blackbars)andmasspervolumeoforiginalsample(graybars).Samplesandtheirrespectivefilteredvolumesareshownonthex-axis.Errorbarsindicatevariabilitybetweenextractionsfromtriplicatesamples.BlackdiamondsshowthecoefficientsofvariationfortheDNArecoveredfromtriplicatemembranefiltersofthesamesample.S4FigureS3.Rankabundancedistributions(averagedoverthreeseasons)fortheOTUsdetectedineachsamplinglocation.Circlesshowthemeanrelativeabundanceoftheleastabundanttaxadetectedinthisstudy.ThemeanrelativeabundanceoftherarestOTUswasbetween0.1-0.12%forallsamplinglocations.Hence,deepersequencingwouldonlyresultingreaterdetectionofraretaxawithmeanrelativeabundanceequaltoorlessthan0.1-0.12%.S5FigureS4.Theleftpanelshowsaneighbor-joining(1,000iterations)treehighlightingthephylogeneticrelationshipamongthe25coreOTUs.TheterminalleafsofthephylogenetictreerepresenttheseOTUslabeledindescendingorderoftheirmeanrelativeabundanceasobservedacrossallsamplinglocations.ThemiddlepanelshowsthemeanabundanceofeachofthecoreOTUsacrossthesixsamplinglocations,andtherightpanelrepresentstheheat-mapofchangesinrelativeabundanceatthesixsamplinglocations.S6FigureS5.Jackknifeclusteringwasperformedwith1,000iterationsforsamplinglocationswithineachseason(A)andforallseasonscombined(B)usingtheWUnFmetric.ThenodecolorsindicatetheJackknifefractions,i.e.,thepercentageofiterationsforwhicheachnodewasconserved.ParsimonypvaluesareindicatedinFigureS6Bforpairwisecomparisonsforfilter-filtereffluent,filtereffluent-disinfectiontank,anddisinfectiontank-DWDSandshowthatthesesamplinglocationshadbacterialcommunitystructuresthatwerenotsignificantlydifferentfromeachother(p0.001).S7TableS1.Asummaryofsamplescollected(n=120)andpoolingstrategyresultinginatotalof18samplesforsequencing.Theleftcolumnshowsthesamplecollected,andtherightcolumnindicatesthefinal,pooledsamplesssequencedforeachseason.CollectedsamplesSequencedsamplesMonthAprJunJulAugSepOctSeasonSpringSummerFallLocationLocationRiverSource-SpringSource-SummerSource-FallSourceWellFilterInfluentFilterinfluent-SpringFilterinfluent-SummerFilterinfluent-FallFilterinfluentFilterFilter-SpringFilter-SummerFilter-FallFilterFiltereffluentFiltereffluent-SpringFiltereffluent-SummerFiltereffluent-FallFiltereffluentClearwellDisinfection-SpringDisinfection-SummerDisinfection-FallDisinfectionReservoirDistributionsystemDistributionsystem-SpringDistributionsystem-SummerDistributionsystem-FallDistributionsystemS8TableS2.Summaryofquality-filteredandchimerafreereadsforeachseasonalsample.SampleSeasonQuality-scorefilteredreadsChimerafreereadsSourceSpring243238Summer358355Fall329326FilterinfluentSpring207204Summer381376Fall417410FilterSpring333327Summer428418Fall171165FiltereffluentSpring322319Summer334331Fall302295DisinfectionSpring316312Summer263257Fall263256DistributionsystemSpring239236Summer351344Fall460454S9TableS3:Asummaryofwaterqualityparametersforthefivebulkwatersamplinglocationsacrossthethreeseasons.Dataareshownasaveragesofreplicateanalyseswithstandarddeviationsinparentheses.ND:Non-detectable.NQ:Detectablebutnotquantifiable.NA:Notapplicable.SpringSummerFallTotalorganiccarbon(mg/l)Source10.4(4.8)5.1(3.4)6.1(3.5)Filterinfluent6.7(0.5)4.6(1.6)4.2(1.8)Filtereffluent5.1(0.2)4.2(0.8)3.4(1.4)Disinfectiontank5.5(0.7)3.7(0.2)3.3(1.4)Distributionsystem6.4(2.6)3.5(0.6)3.7(0.6)pHSource7.6(0.4)7.8(0.4)7.9(0.4)Filterinfluent9.2(0.2)9.5(0)9.4(0.2)Filtereffluent9.0(0.2)9.3(0.1)9.4(0.2)Disinfectiontank9.1(0.1)9.3(0.1)9.3(0.2)Distributionsystem9.1(0.1)9.3(0.1)9.3(0.1)Ammonia(µg-N/l)Source59(85)63(39)8(16)Filterinfluent123(14)134(17)51(2)Filtereffluent17(10)11(3)NDDisinfectiontank387(91)304(58)305(79)Distributionsystem474(294)334(68)253(68)Nitrite(µg-N/l)*NQ10µg-N/lSource249(490)NQ*NDFilterinfluentNQ*NQ*NDFiltereffluent50(50)NDNQ*DisinfectiontankNQ*NQ*NDDistributionsystem10(9)45(9)NQ*Nitrate(µg-N/l)*NQ20µg-N/lSourceNQ*230(160)191(286)FilterinfluentNQ*258(258)494(70)FiltereffluentNQ*209(56)709(312)DisinfectiontankNQ*197(163)590(481)DistributionsystemNQ*250(224)264(224)S10Phosphate(µg-N/l)*NQ50µg-P/lSourceNDNDNDFilterinfluent77(42)39(30)59(59)Filtereffluent57(4)67(12)92(71)Disinfectiontank53(45)96(40)936(36)Distributionsystem153(41)148(60)99(60)Temperature(°°°°C)Source16(3)21(4)17(3)Filterinfluent16(3)23(1)15(3)Filtereffluent17(2)23(0)15(4)Disinfectiontank18