两种不同类型微生物源示踪技术的比较与应用

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1、山东大学硕士学位论文对水样中的E．coli分别进行库依赖法源示踪和非库依赖法源示踪，结果均表明非点源粪便污染主要为未知来源，贡献率约为30％(非库依赖法)到70％(库依赖法)，其次为人源，再次为鸡源与犬源。结论本课题结果表明基于REP．PCR法的微生物源示踪技术具有较高的可靠性，在不同类型的源解析方法下源示踪准确率绝大多数高于60％，特别是库依赖的源解析方法下源示踪准确率表现优秀，对于同一地区样本可达94％以上，对于不同地区样本在85％以上。库依赖法受到地理性差异影响小于非库依赖法，适宜更大范围的应用

2、。同时结果还表明小清河沿岸的非点源污染类型除未知来源外主要为人类粪便，其次为鸡源与犬源，这一结论与当地实际情况较为符合。本课题研究结果为下一步微生物源示踪技术的研发以及应用提供了参考。关键词：水污染：环境监测：水微生物学：微生物源示踪技术：非点源粪便污染4山东大学硕士学位论文Comoarisonandaoolicationoftwo‘ti～rentkindsof1。icrobiaomlaarlsonantwolllerentslVilCrODia’l‘0ISourcetrackingAbstractM

3、icrobialSourceTracking(MST)isanewtechniquebasingontheassumptionthatbymonitoringthemicrobialindicatorswithmolecularandbiochemicalmethods，theoriginofpathogenicmicroorganismsCanbetrackedandcontrolled．MSTcanalsoassessthecontributionsofthevariouscontaminatio

4、nsources．Itissupposedtobeaneffectiveapproachtocontrolnon-pointfecalcontamination．Inthisissue，twolibrary—dependentandlibraryindependentmethodsarecomparedandapplied．Thisstudycanprovideatheoreticalbasisforfurtherresearchandapplicationreferenceformonitoring

5、andmanagingofnon—pointsourcefecalpollution．ObjectThefecalsamplesofdifferentsourceswerecollectedfromDalianandJinan．Bytrackingthepollutionsourceswithlibrary··dependentandlibrary-·independentmicrobialsourcetrackingmethods，theaccuracyandstabilityofthesetwok

6、indsofmethods，especiallytheimpactofgeographicvariationwerecompared．Theapplicationvalueofmicrobialsourcetrackingwasanalysedbytrackingthecontaminationsourcesofwatersampleswithbothoflibrary—dependentandlibrary-independentmethodsandthecontributionofdifferen

7、tcontaminationsourceswereassessedMethodsDalianandJinanweresetassamplingpoints．Watersamplesandfecalsamplesfromchicken，dog，cattle，pigandhumansourceswerecollectedbysterileoperation．TheATCC25922wereusedasstandardE．coliinthisissue．Eosinmethyleneblue(EMB)test

8、s，sugarfermentationtests，IMVICtestswereusedtoisolateandidentifyE．colifromthesamples．DNAwereextractedfromE．colisamplesbyEasyPureGenomicDNAExtractionKitandtestedbyUVspectrophotometer．OD600．OD260andOD280wereusedtotestwhethertheDNAis