Human source identification by using a human-associated Escherichia coli genetic marker in the Mae Klong River, Thailand.

Contamination caused by microbial fecal pollution in water bodies is a serious problem in many countries, especially in low- and middle-income countries. Therefore, fecal source tracking is an important method used to understand the source of fecal contamination and to decrease the hazard of waterborne diseases that occurs in the environment. In this study, a human-associated genetic marker for Escherichia coli (H8) was used to investigate the source of fecal contamination in the Mae Klong River, Thailand. Real-time PCR was performed with this marker for 500 E. coli isolates collected from 10 sampling sites along the river, including MK10 (upstream) to MK1 (downstream). The results showed that the proportions of H8-positive isolates were 46, 14, 10, 18, 14, 38, 12, 26, 32, and 14% at MK10-MK1, respectively. All positive proportions were significantly different between the locations (p < 0.001). The higher occurrence of E. coli with H8 marker detection indicated that domestic wastewater was largely discharged without proper treatment, which is attributable to the high population and the absence of proper sewage treatment in those areas.

Evaluation of a human-associated genetic marker for Escherichia coli (H8) for fecal source tracking in Thailand.

High levels of microbial fecal pollution are a major concern in many countries. A human-associated genetic marker for Escherichia coli (H8) has recently been developed for fecal source tracking. The assessment of the H8 marker performance is crucial before it can be applied as a suitable method for fecal source tracking in each country. The performance (specificity and sensitivity) of the H8 marker was evaluated by using non-target host groups (cattle, buffalo, chicken, duck, and pig feces) and target host groups (influent and effluent from a wastewater treatment plant and septages). SYBR based real-time PCR (polymerase chain reaction) was done on 400 E. coli isolates from non-target and target host groups after E. coli isolation. It was found that the specificity from animal feces samples collected in Thailand was 96%. Moreover, influent, effluent, and septage samples showed the values of the sensitivity at 18, 12, and 36%, respectively. All of the non-target host groups were found to be significantly different with positive proportions from the target host group (septage samples) (p ≤ 0.01). Based on the results, this marker is recommended for use as a human-associated E. coli marker for identifying sources of fecal pollution in Thailand.