Gregory T. Kleinheinz

Effects of the nuisance algae, Cladophora, on Escherichia coli at recreational beaches in Wisconsin

Recreational beaches constitute a large part of the 12 billion dollar per year tourism industry in Wisconsin. Beach closures due to microbial contamination are costly in terms of lost tourism revenue and adverse publicity for an area. Escherichia coli (E. coli), is used as an indicator of microbial contamination, as high concentrations of this organism should indicate a recent fecal contamination event that may contain other, more pathogenic, bacteria. An additional problem at many beaches in the state is the nuisance algae, Cladophora. It has been hypothesized that mats of Cladophora may harbor high concentrations of E. coli. Three beaches in Door County, WI were selected for study, based on tourist activity and amounts of algae present. Concentrations of E. coli were higher within Cladophora mats than in surrounding water. Beaches displayed an E. coli concentration gradient in water extending away from the Cladophora mats, although this was not statistically significant. Likewise, the amount of Cladophora observed on a beach did not correlate with E. coli concentrations found in routine beach monitoring samples. More work is needed to determine the impact of mats of Cladophora on beach water quality, as well as likely sources of E. coli found within the mats.

The Green Alga, Cladophora , Promotes Escherichia coli Growth and Contamination of Recreational Waters in Lake Michigan

A linkage between Cladophora mats and exceedances of recreational water quality criteria has been suggested, but not directly studied. This study investigates the spatial and temporal association between Escherichia coli concentrations within and near Cladophora mats at two northwestern Lake Michigan beaches in Door County, Wisconsin. Escherichia coli concentrations in water underlying mats were significantly greater than surrounding water (p < 0.001). Below mat E. coli increased as the stranded mats persisted at the beach swash zone. Water adjacent to Cladophora mats had lower E. coli concentrations, but surpassed EPA swimming criteria the majority of sampling days. A significant positive association was found between E. coli concentrations attached to Cladophora and in underlying water (p < 0.001). The attached E. coli likely acted as a reservoir for populating water underlying the mat. Fecal bacterial pathogens, however, could not be detected by microbiological culture methods either attached to mat biomass or in underlying water. Removal of Cladophora mats from beach areas may improve aesthetic and microbial water quality at affected beaches. These associations and potential natural growth of E. coli in bathing waters call into question the efficacy of using E. coli as a recreational water quality indicator of fecal contaminations.

Impact of the Alga Cladophora on the Survival of E. coli, Salmonella, and Shigella in Laboratory Microcosm

ABSTRACT E. coli is an indicator of recent fecal contamination of freshwater beaches around the Great Lakes region. Elevated concentrations indicate that a fecal contamination has occurred, and that the risk for contact with fecal pathogenic organisms is heightened. The green algae, Cladophora, harbors populations of E. coli and potentially allows for prolonged survival and even replication of the bacterium in the lake environment. If presence of Cladophora mats on beaches is associated with persistence of E. coli in beach water, then E. coli would be a useful indicator organism only if pathogens also were able to survive and persist in the algae. This study utilized lab microcosms to study the persistence of E. coli, and of the fecal pathogens, Salmonella and Shigella, in lake water with and without the presence of Cladophora. E. coli was able to persist for extended periods in the presence of Cladophora (attached to algal mats for 45 days). Salmonella and Shigella, however, were unable to persist for this time period while in the presence of Cladophora (Salmonella attached to Cladophora was detectable for 10 days and Shigella was detectable for only 2 days). These data imply that E. coli is able to survive in the presence of Cladophora for greater times than are the fecal pathogens and that E. coli may not be an appropriate indicator organism for beaches with accumulations of algal material.

Multi-scale temporal and spatial variation in genotypic composition of Cladophora-borne Escherichia coli populations in Lake Michigan

High concentrations of Escherichia coli in mats of Cladophora in the Great Lakes have raised concern over the continued use of this bacterium as an indicator of microbial water quality. Determining the impacts of these environmentally abundant E. coli, however, necessitates a better understanding of their ecology. In this study, the population structure of 4285 Cladophora-borne E. coli isolates, obtained over multiple three day periods from Lake Michigan Cladophora mats in 2007-2009, was examined by using DNA fingerprint analyses. In contrast to previous studies that have been done using isolates from attached Cladophora obtained over large time scales and distances, the extensive sampling done here on free-floating mats over successive days at multiple sites provided a large dataset that allowed for a detailed examination of changes in population structure over a wide range of spatial and temporal scales. While Cladophora-borne E. coli populations were highly diverse and consisted of many unique isolates, multiple clonal groups were also present and accounted for approximately 33% of all isolates examined. Patterns in population structure were also evident. At the broadest scales, E. coli populations showed some temporal clustering when examined by year, but did not show good spatial distinction among sites. E. coli population structure also showed significant patterns at much finer temporal scales. Populations were distinct on an individual mat basis at a given site, and on individual days within a single mat. Results of these studies indicate that Cladophora-borne E. coli populations consist of a mixture of stable, and possibly naturalized, strains that persist during the life of the mat, and more unique, transient strains that can change over rapid time scales. It is clear that further study of microbial processes at fine spatial and temporal scales is needed, and that caution must be taken when interpolating short term microbial dynamics from results obtained from weekly or monthly samples.

Effects of Rainfall on E. coli Concentrations at Door County, Wisconsin Beaches.

Rainfall and its associated storm water runoff have been associated with transport of many pollutants into beach water. Fecal material, from a variety of animals (humans, pets, livestock, and wildlife), can wash into beach water following rainfall and result in microbial contamination of the beach. Many locales around the world issue pre-emptive beach closures associated with rainfall. This study looked at eight beaches located in Door County, Wisconsin, on Lake Michigan to determine the impact of rainfall on E. coli concentrations in beach water. Water samples were collected from beach water and storm water discharge pipes during rainfall events of 5u2009mm in the previous 24 hours. Six of the eight beaches showed a significant association between rainfall and elevated beach water E. coli concentrations. The duration of the impact of rainfall on beach water E. coli concentrations was variable (immediate to 12 hours). Amount of rainfall in the days previous to the sampling did not have significant impact on the E. coli concentrations measured in beach water. Presence of storm water conveyance pipes adjacent to the beach did not have a uniform impact on beach water E. coli concentrations. This study suggests that each beach needs to be examined on its own with regard to rain impacts on E coli concentrations in beach water.

Evaluation of Avian Waste and Bird Counts as Predicators of Escherichia coli Contamination at Door County, Wisconsin Beaches

ABSTRACT Microbial source tracking (MST) has become a focus of some recreational beach monitoring programs. Suspected sources of contamination include human sewage, agricultural runoff, and feces from wildlife and domestic animals, depending on beach location. Waterfowl have been suggested as a primary source of fecal contamination at many beaches, but techniques to “prove” contaminating microbes are of avian origin are mostly unsubstantiated. Researchers often rely on bird counts to measure the impact of waterfowl on beach health. Since waterfowl populations at Door County, Wisconsin (USA) beaches are transitory, this study focused on enumeration of avian waste material along beach transects, rather than on once per day “snapshot” bird counts. Escherichia coli (E. coli) concentration in beach water was not correlated with avian waste counts at the ten beaches studied in 2004 or the 13 studied in 2005 (rural to semi-urban). Bird counts correlated with E. coli concentrations in beach water at 30% of the sample sites in 2004 and at only one site in 2005. During the 2004 swimming season avian waste counts correlated with bird counts at only one beach and there was no correlation in 2005. These results indicate that neither avian waste enumeration nor bird counts can successfully be used to predict microbial contamination of recreational water at selected Great Lakes beaches.

E. coli at Lake Superior Recreational Beaches

Increased usage of public beaches and heightened awareness of the need to monitor water for potential microbial contamination have led to passage of the U.S. Beaches Environmental Assessment and Coastal Health Act (BEACH Act) in 2000. This legislation calls for bacterial monitoring of recreational waters along the U.S. coastline, including the Great Lakes. The State of Wisconsin implemented this legislation in summer 2003, triggering extensive microbial monitoring of Lake Superior beaches. E. coli were measured at 27 beaches along Lake Superior, Wisconsin by defined substrate analysis. While E. coli concentrations were relatively low at these “cold water” beaches, monitoring did result in a few swimming advisories and beach closures (0.8% of total samples collected had E. coli concentrations that exceeded standards). Increasing water temperatures were not associated with increasing concentrations of bacterial contaminants. Location of sampling site and depth of water at the location where samples were taken did have an effect on detection of E. coli. Greater E. coli levels consistently were detected in shallower water and varied depending on the location of the sampling site horizontally across the beach. These findings support the notion that the concentration of E. coli in chest deep water may not be representative of E. coli concentrations in shallower water, closer to shore.

Microbial Concentrations in Sand and their Effect on Beach Water in Door County, Wisconsin

ABSTRACT The seasonal variations and patterns of Escherichia coli in Wisconsins coastal waters have been closely studied in recent years due to increased beach monitoring activities. Patterns of distribution of the indicator organism, E. coli, in the sand at these beaches are now being investigated as a source of E. coli to adjacent beach water. This project investigates the concentrations of E. coli in beach sand, and the relationship between these sand-microbe concentrations and concentrations of microbes in the corresponding beach water. Weekly sampling of upshore, swash, and submerged sand at six beaches provided numbers of the indicator bacteria in each beachs sand substrate for two consecutive summers. Overall concentrations of E. coli were highest in the swash sand of the beach, with the highest numbers seen in the summer months and lowest numbers in the winter months. Each location had very different concentrations of E. coli in the beach sand from 1,800 CFU/100 g to 21,670 CFU/100 g sand. Each location had a very different relationship between the indicator organism found in the beach sand and that found in the beach water. These data suggest that sand may be a reservoir for E. coli at some locations, and another source of contamination that should be considered in beach monitoring programs. However, elevated levels of E. coli in beach sand were not universal and varied greatly from location to location.

Human and Bovine Viruses and Bacteria at Three Great Lakes Beaches: Environmental Variable Associations and Health Risk

Waterborne pathogens were measured at three beaches in Lake Michigan, environmental factors for predicting pathogen concentrations were identified, and the risk of swimmer infection and illness was estimated. Waterborne pathogens were detected in 96% of samples collected at three Lake Michigan beaches in summer, 2010. Samples were quantified for 22 pathogens in four microbial categories (human viruses, bovine viruses, protozoa, and pathogenic bacteria). All beaches had detections of human and bovine viruses and pathogenic bacteria indicating influence of multiple contamination sources at these beaches. Occurrence ranged from 40 to 87% for human viruses, 65-87% for pathogenic bacteria, and 13-35% for bovine viruses. Enterovirus, adenovirus A, Salmonella spp., Campylobacter jejuni, bovine polyomavirus, and bovine rotavirus A were present most frequently. Variables selected in multiple regression models used to explore environmental factors that influence pathogens included wave direction, cloud cover, currents, and water temperature. Quantitative Microbial Risk Assessment was done for C. jejuni, Salmonella spp., and enteroviruses to estimate risk of infection and illness. Median infection risks for one-time swimming events were approximately 2 × 10(-5), 8 × 10(-6), and 3 × 10(-7) [corrected] for C. jejuni, Salmonella spp., and enteroviruses, respectively. Results highlight the importance of investigating multiple pathogens within multiple categories to avoid underestimating the prevalence and risk of waterborne pathogens.

Effect of aquatic macrophytes on the survival of Escherichia coli in a laboratory microcosm

Abstract Kleinheinz, G., A. Coenan, T. Zehms, J. Preedit, M.C. Leewis, D. Becker and C. McDermott. 2009. Effect of aquatic macrophytes on the survival of Escherichia coli in a laboratory microcosm. Recreational beaches are very important as engines for tourism revenue in many areas of Wisconsin, United States, and microbial contamination of beach water can be very costly. Escherichia coli (E. coli) is the indicator organisms used in the Great Lakes region of the United States for recreational water monitoring. The overall objective of this study was to determine if aquatic macrophytes (Sagittaria sp., Myriophyllum sp.) allow for prolonged E. coli survival in the environment by using a laboratory microcosm as a test system. This study tested whether E. coli survival was greater in lake water alone or in lake water containing a high density of macrophytes (HDM), a low density of macrophytes (LDM), or plastic plant material. Water and plant substrate samples were analyzed for E. coli concentrations. ANOVA analysis indicated a significant difference between all treatments (p < 0.001). The Scheffe test revealed a significant difference between E. coli concentrations in water in the HDM treatment and the plastic plants (p = 0.003) and the controls (lake water only; p < 0.001), as well as differences between the LDM treatments and the plastic plants (p < 0.001) and the controls (p < 0.001). Attachment of E. coli to the macrophytes was rapid and may contribute to the rapid decline of E. coli found in the microcosm water samples. The presence of aquatic macrophytes did not appear to provide growth factors or other substances that could prolong the survival of E. coli in water, but the plants may contribute to bacterial survival by providing a medium for bacterial attachment. To our knowledge this is the first study reporting on the effects of aquatic macrophytes on E coli concentrations in water.