Discrimination of human and non-human fecal sources with rapid methods in coastal waters and sediments
- Author(s): Zimmer-Faust, Amity Gayle
- Advisor(s): Jay, Jennifer A
- et al.
Microbial contamination in coastal waters is an important public health and economic problem worldwide. It is common for the sources as well as the environmental fate and transport of fecal contamination in recreational waters to be unknown making it difficult to mitigate the input and to understand the relative health risk associated with a given water body. A comprehensive understanding of sources and bacterial dynamics is needed for effective mitigation and management of microbial contaminants.
The main objectives of this study were to identify sources of fecal contamination to a chronically impaired Southern California watershed, to demonstrate applicability of microbial source tracking (MST) tools, including source-specific markers for discrimination of human and non-human sources, to evaluate the fate of fecal contaminates in coastal sediments, and to develop and optimize immunomagnetic separation/adenosine triphosphate (IMS/ATP) assays for rapid enumeration of viable fecal contamination.
A three-year MST study was conducted to help explain elevated levels of surfzone fecal indicator bacteria (FIB) at Topanga State Beach, a critically impaired Southern California Beach. This study investigated sources of FIB to the Topanga watershed and the applicability of using MST technology longer time scales. MST markers effectively elucidated temporal and seasonal trends in fecal bacterial levels, and dog and gull marker appeared to be a significant sources to Topanga lagoon and Topanga State Beach. However, a lack of correlation between FIB and marker measurements was noted, and dog marker and FIB levels did not covary when compared at different Southern California beaches.
Sediments were found to play an important and variable role in environmental fate of MST markers and FIB. Variable decay was observed for different indicators and in different sediments, with differences noted even within one watershed. The human HF183 marker was useful for providing evidence of recent inputs of human fecal contamination and behaved similarly to the molecular marker for Campylobacter (qCAMP) and FIB in brackish sediments. The general Bacteroides (GB3) and enterococci (ENT1A) markers were more conservative and under certain circumstances had comparable decay to culturable FIB. Application of a suite of markers may be necessary for effective evaluation of sediment fecal bacterial levels. Moreover, differences were observed between relative decay amongst the different sediments tested, illustrating the need for more routine sediment monitoring.
IMS/ATP assays provided useful information regarding fecal contamination levels and measurements made had a consistent relationship with measurements made by standard methods. IMS/ATP utilizes paramagnetic beds and target-specific antibodies to isolate target organisms. Following isolation, adenosine tri-phosphate (ATP) is extracted from the target population and quantified. The Cov-IMS/ATP method rapidly measured viable enterococci in complex surface waters, providing a useful ﬁeld tool for assessment of coastal water quality and for identiﬁcation of hot spots of fecal contamination. An inversely-coupled (Inv-IMS/ATP) assay for detection of Bacteroides thetaiotaomicron was developed and applied for rapid detection of human-associated fecal contamination. The Inv-IMS/ATP assay yielded measurements of viable B. thetaiotaomicron that were comparable to the HF183 human marker in complex source waters impacted with both wastewater and runoff, and the Inv-IMS/ATP assay was able to effectively differentiate between surface waters impacted with adequately and inadequately treated wastewater. IMS/ATP assays show promise for rapid evaluation of recreational water quality in areas where access to more expensive methods is limited and in areas where water quality is unpredictable.
This research highlights the difficulties and complexities associated with effective tracking and management of microbial contaminates in the coastal environment. Additional research evaluating relative aging of molecular markers and relative contributions from different sources is needed to fully interpret field-based source marker data. Sediments were shown to have an important and variable role in fate of fecal contaminants in the environment. Additional studies are needed evaluating how watershed models can most effectively be adapted to include a sediment compartment and how different sources of fecal contamination and source markers decay in sediments with variable characteristics. IMS/ATP assays showed promise and can be successfully applied in complex waters for rapid enumeration of viable fecal contamination; additional verification of assay performance is needed at complex sites impacted with multiple sources.