Implementation and Integration of Microbial Source Tracking in a River Watershed Monitoring Plan

Fecal pollution of water bodies poses a serious threat for public health and ecosystems. Microbial source tracking (MST) using host specific bacteria are used to track the source of this potential pollution and be able to perform a better management of the pollution at the source. In this study we tested 12 molecular MST markers to track human, ruminant, sheep, horse, pig and gull pollution to determine their usefulness in their application for an effective management of water quality. First, the potential of the selected markers to track the source was evaluated using fresh fecal samples. Subsequently, we evaluated their performance in a catchment with different impacts, considering land use and environmental conditions. All MST markers showed high sensitivity and specificity, although none achieved 100% for both. Although some of the MST markers were detected in hosts other than the intended ones, their abundance in the target group was always several orders of magnitude higher than in the non-target hosts, demonstrating their suitability to distinguish between sources of pollution. The MST analysis matched the land use in the watershed allowing a very accurate assessment of the main hazards and sources of pollution, in this case mainly human and ruminant pollution. Correlating environmental parameters like temperature and rainfall with the levels of the MST markers provided insight into the dynamics of the pollution along the catchment. The levels of the human associated marker showed a significant negative correlation with rainfall in human polluted areas suggesting a dilution of the pollution, whereas at agricultural areas the ruminant marker increased with rainfall. There were no seasonal differences in the levels of human marker, indicating human pollution as a constant pressure throughout the year, whereas the levels of the ruminant marker was influenced by the seasons, being more abundant in summer and autumn. Performing MST analysis integrated with land uses and environmental data can improve the management of fecal polluted areas and set up good practices.