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Validation of Bacteroidales-based microbial source tracking markers for Pig fecal pollution and their application in two rivers of North China

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Frontiers of Environmental Science & Engineering 2020年第4期14卷 139-146页

作者： Youfen Xu Zong Li Ruyin Liu Hongxia Liang Zhisheng Yu Hongxun Zhang College of Resources and Environment University of Chinese Academy of SciencesBeijing 100049China

In China,pig feces is the predominant source of excrement produced b y animal husbandry.Improper use or direct discharge of pig feces can result in contamination of natural water systems.Microbial source tracking(MST)technology can identify the sources of fecal pollution in environmental water,and contribute to the management of Pig fecal pollution by local environmental protection agencies.However,the accuracy of such assays can be context-dependent,and they have not been comprehensively evaluated under Chinese conditions.We aimed to compare the performance of five previously reported nig-specific MST assays(PF,pig-Bac 1 STBR,pig-Bac2 SYBR,pig-1-Bac TaqMan,and pig-2-Bac TaqMan,which are based on Bacteroidales 16S rRNA gene markers)and apply them in two rivers of North China.We collected a total of 173 fecal samples from pigs,cows,goats,chickens,humans,and horses across China.The PF assay optimized in this study showed outstanding qualitative performance and achieved 100%specificity and sensitivity.However,the two SYBR green qPCR assays(pig-Bac1 SYBR and pig-Bac2 SYBR)cross-reacted with most non-pig fecal samples.In contrast,both the pig-l-Bac TaqMan and pig-2-Bacr TaqMan assays gave 100%specificity and sensitivity.Of these,the pig-2-Bac TaqMan assay showed higher reproducibility.Our results regarding the specificity of these pig-specific MST assays differ from those reported in Thailand,Japan,and America.Using the PF and pig-2-Bac TaqMan assays,a field test comparing the levels of Pig fecal pollution in rivers near a pig farm before and after comprehensive environmental pollution governance indicated that Pig fecal pollution was effectively controlled at this location.

关键词： Microbial source tracking Pig fecal pollution 16S rRNA genem arkers pig-specific Bacteroidales

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