Quantitative assessment of bacterial pathogens, their geographic variability, and distribution in various matrices at Great Lakes beaches are limited. Quantitative PCR (qPCR) was used to test for genes from E. coli O157:H7 (eaeO157), shiga-toxin producing E. coli (stx2), Campylobacter jejuni (mapA), Shigella spp. (ipaH), and a Salmonella enterica-specific (SE) DNA sequence at seven Great Lakes beaches, in algae, water, and sediment. Overall, detection frequencies were mapA>stx2>ipaH>SE>eaeO157. Results were highly variable among beaches and matrices; some correlations with environmental conditions were observed for mapA, stx2, and ipaH detections. Beach seasonal mean mapA abundance in water was correlated with beach seasonal mean log10 E. coli concentration. At one beach, stx2 gene abundance was positively correlated with concurrent daily E. coli concentrations. Concentration distributions for stx2, ipaH, and mapA within algae, sediment, and water were statistically different (Non-Detect and Data Analysis in R). Assuming 10, 50, or 100% of gene copies represented viable and presumably infective cells, a quantitative microbial risk assessment tool developed by Michigan State University indicated a moderate probability of illness for Campylobacter jejuni at the study beaches, especially where recreational water quality criteria were exceeded. Pathogen gene quantification may be useful for beach water quality management.

Better understanding of Escherichia coli population dynamics and genetic variability in the secondary habitat is essential to improve fecal contamination monitoring and contamination pathway characterization. In this study, water samples were collected monthly over a one-year period at eight locations in the Catoma Creek watershed, a mixed land-use watershed in CentralAlabama. E. coli concentrations varied from 17 to 12,650 CFU/100 ml and were well correlated with stream flow rates. Repetitive sequence-based PCR DNA fingerprinting was used to generate 271 unique DNA fingerprint patterns from 502 E. coli isolated from water samples. Cluster analysis showed an overall similarity of 32.8% across all DNA fingerprints. Multivariate analysis of variance (MANOVA) showed that E. coli genotypes had a tendency to cluster according to season and stream flow rather than sampling sites. MANOVA of a subset of data within a given season and flow rate, however, revealed some geographical differentiation between urban and rural sampling sites. The results indicate that genetic diversity of E. coli populations was not only high in the secondary habitat but also varied with season, flow conditions and, to a lesser extent, sampling location. To our knowledge, this is the first report relating E. coli genotype to stream flow.

c Clostridium botulinum type E toxin is responsible for extensive mortality of birds and fish in the Great Lakes. The C. botulinum bontE gene that produces the type E toxin was amplified with quantitative PCR from 150 sloughed algal samples (primarily Cladophora species) collected during summer 2012 from 10 Great Lakes beaches in five states; concurrently, 74 sediment and 37 water samples from four sites were also analyzed. The bontE gene concentration in algae was significantly higher than in water and sediment (P < 0.05), suggesting that algal mats provide a better microenvironment for C. botulinum. The bontE gene was detected most frequently in algae at Jeorse Park and Portage Lake Front beaches (Lake Michigan) and Bay City State Recreation Area beach on Saginaw Bay (Lake Huron), where 77, 100, and 83% of these algal samples contained the bontE gene, respectively. The highest concentration of bontE was detected at Bay City (1.98 ؋ 10 5 gene copies/ml of algae or 5.21 ؋ 10 6 g [dry weight]). This study revealed that the bontE gene is abundant in the Great Lakes but that it has spatial, temporal, and matrix variability. Further, embayed beaches, low wave height, low wind velocity, and greater average water temperature enhance the bontE occurrence.

Fecal pollution is the leading cause of beach closures in coastal regions of the United States. Identification of fecal pollution sources is necessary to implement effective pollution control strategies. This study sought to determine whether humans and cattle contributed to the fecal contamination at a public beach in Alabama, USA. High levels of enterococci were observed after significant rainfall events, of which 36 samples (35.6%) exceeded USEPA's statistical threshold value of 130 CFU/100 mL. Quantitative polymerase chain reaction (qPCR) assays were used to determine general (AllBac), human-(HF183), and cattle-associated (CowM3) Bacteroidales. There was a positive correlation between the levels of enterococci and AllBac markers. Our results suggest that fecal contamination from cattle was minimal and contamination from humans appeared to originate from diffuse sources. Stormwater runoff has the potential to supply considerable loads of fecal bacteria to coastal waters. Further research is needed to determine the contribution of other animals to fecal contamination in the region. 84 Water and Environment Journal 32 (2018) 84-93 V C 2017 CIWEM. Water and Environment Journal. Print ISSN 1747-6585 J. Xue et al. Fecal pollution sources in coastal waters Fecal pollution sources in coastal waters J. Xue et al.