Assessment and application of host-specific Bacteroidales genetic markers for microbial source tracking of river water in Japan

Microbial source tracking using host-specific microbial genetic markers is considered a promising approach to determine fecal contamination sources of aquatic environments. This study aimed to assess the application of previously developed host-specific Bacteroidales quantitative PCR assays to microbial source tracking of river water samples in Yamanashi Prefecture, Japan. Various types of fecal-source samples, such as raw sewage, secondary-treated sewage of a wastewater treatment plant, and cattle feces, were used for three human-, two ruminant- and two pig-specific Bacteroidales quantitative PCR assays. Our results demonstrated that BacHum, BacR and Pig2Bac assays as suitable human-, ruminant- and pig-specific assays, with an accuracy of 86%, 94% and 77%, respectively. These selected assays were used for microbial source tracking of 63 river water samples collected at nine sites in two river basins. From these sites, there were 48 (76%), 34 (54%) and 9 (14%) positive samples using the BacHum, BacR and Pig2Bac assays, respectively. These assays revealed the effects of humans and animals on fecal contamination of river water.