Assessment of mixed human, livestock and wildlife sources of fecal contamination in surface waters by mitochondrial DNA metagenomics

Fecal contaminations of surface waters and groundwater can occur by more than one animal species in areas where urban and agricultural activities overlapped. We use mitochondrial metagenomics for source tracking fecal contamination in 86surface water samples. These PCR primers target the mitochondrial 16S ribosomal RNA genes from mammals, birds and fishes, and majority of amphibians or reptiles. These primers were used to amplify mitochondrial DNA (mtDNA) among eDNA in surface waters to identify homeotherms as a potential source of fecal pollution. The amplicons were sequenced by the Illumina MiSeq PE-250 platform.

在城市与农业活动交汇的区域，地表水与地下水的粪便污染可由多种动物物种引发。本研究采用线粒体宏基因组学（mitochondrial metagenomics）对86份地表水样本中的粪便污染来源进行追踪。本次使用的PCR引物可靶向扩增哺乳类、鸟类、鱼类以及多数两栖类或爬行类动物的线粒体16S核糖体RNA基因，通过该引物对地表水中的环境DNA（eDNA）内的线粒体DNA（mtDNA）进行扩增，以鉴定恒温动物作为粪便污染的潜在来源。扩增产物通过Illumina MiSeq PE-250测序平台完成测序。