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.

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.

Use of microarray hybridization to identify Brugia genes involved in mosquito infectivity.

Brugia malayi and Brugia pahangi microfilariae (mf) require a maturation period of at least 5 days in the mammalian host to successfully infect laboratory mosquitoes. This maturation process coincides with changes in the surface composition of mf that likely are associated with changes in gene expression. To test this hypothesis, we verified the differential infectivity of immature (< or =3 day) and mature (>30 day) Brugia mf for black-eyed Liverpool strain of Aedes aegypti and then assessed transcriptome changes associated with microfilarial maturation by competitively hybridizing microfilarial cDNAs to the B. malayi oligonucleotide microarray. We identified transcripts differentially abundant in immature (94 in B. pahangi and 29 in B. malayi) and mature (64 in B. pahangi and 14 in B. malayi) mf. In each case, >40% of Brugia transcripts shared no similarity to known genes or were similar to genes with unknown function; the remaining transcripts were categorized by putative function based on sequence similarity to known genes/proteins. Microfilarial maturation was not associated with demonstrable changes in the abundance of transmembrane or secreted proteins; however, immature mf expressed more transcripts associated with immune modulation, neurotransmission, transcription, and cellular cytoskeleton elements, while mature mf displayed increased transcripts potentially encoding hypodermal/muscle and surface molecules, e.g., cuticular collagens and sheath components. The results of the homologous B. malayi microarray hybridization were validated by quantitative reverse transcriptase polymerase chain reaction. These findings preliminarily lend support to the underlying hypothesis that changes in microfilarial gene expression drive maturation-associated changes that influence the parasite to develop in compatible vectors.

Geographic relatedness and predictability of Escherichia coli along a peninsular beach complex of Lake Michigan.

To determine more accurately the real-time concentration of fecal indicator bacteria (FIB) in beach water, predictive modeling has been applied in several locations around the Great Lakes to individual or small groups of similar beaches. Using 24 beaches in Door County, Wisconsin, we attempted to expand predictive models to multiple beaches of complex geography. We examined the importance of geographic location and independent variables and the consequential limitations for potential beach or beach group models. An analysis of Escherichia coli populations over 4 yr revealed a geographic gradient to the beaches, with mean E. coli concentrations decreasing with increasing distance from the city of Sturgeon Bay. Beaches grouped strongly by water type (lake, bay, Sturgeon Bay) and proximity to one another, followed by presence of a storm or creek outfall or amount of shoreline enclosure. Predictive models developed for beach groups commonly included wave height and cumulative 48-h rainfall but generally explained little E. coli variation (adj. R2=0.19-0.36). Generally low concentrations of E. coli at the beaches influenced the effectiveness of model results presumably because of low signal-to-noise ratios and the rarity of elevated concentrations. Our results highlight the importance of the sensitivity of regressors and the need for careful methods evaluation. Despite the attractiveness of predictive models as an alternative beach monitoring approach, it is likely that FIB fluctuations at some beaches defy simple prediction approaches. Regional, multi-beach, and individual beach predictive models should be explored alongside other techniques for improving monitoring reliability at Great Lakes beaches.

Bacteroidales diversity in ring-billed gulls (Laurus delawarensis) residing at Lake Michigan beaches.

This study investigated the occurrence and diversity of Bacteroidales fecal bacteria in gulls residing in the Great Lakes region. Members of this bacterial order have been widely employed as human and bovine host-specific markers of fecal pollution; however, few studies have focused on gulls, which can be a major source of fecal indicator bacteria and pathogens at beaches. We found a low but consistent occurrence of Bacteroidales in gulls at five beaches in three different counties spanning the Wisconsin shoreline of Lake Michigan. The percentages of gulls positive for Bacteroidales were 4 to 8% at beaches in the southern part of the state and 8 to 50% at beaches in the north. Sequencing of 931 clones from seven gull Bacteroidales 16S rRNA gene libraries revealed a large amount of diversity in both individual and pooled gull fecal samples. Two libraries constructed from pooled gull fecal samples (n = 5 and n = 6) did not have a greater richness of sequences than individual samples, suggesting that even within a single gull diversity is high and an extensive sequencing effort is needed to characterize the populations. Estimates of the numbers of operational taxonomic units (OTUs) for the libraries obtained using different similarity levels revealed a large amount of microdiveristy with a limited number of OTUs at the 95% similarity level. Gull sequences were clustered by the beach from which they were collected, suggesting that there were geographic effects on the distribution of Bacteriodales. More than 53% of the 16S rRNA gene sequences from gulls at the southern beaches were associated with the family Porphyromonadaceae, primarily the genus Parabacteroides, whereas sequences from gulls at the northern beaches were comprised of Bacteroidaceae and Prevotellaceae sequences. Comparison of gull sequences with sequences from goose, canine, raccoon, and sewage sources revealed distinct clusters of closely related gull sequences; however, these sequences were widely dispersed across a dendrogram that included all other sources, including previously characterized gull Bacteroidales from other studies, suggesting that geographic influence or simply sample representation plays a greater role in the observed population structure than strictly the host gut environment.

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 5 mm 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.

Discrimination and characterization of environmental strains of Escherichia coli by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

A rapid and reliable bacterial source tracking (BST) method is essential to counter risks to human health posed by fecal contamination of surface waters. Genetic fingerprinting methods, such as repetitive sequence based-PCR (rep-PCR), have shown promise as BST tools but are time-consuming and labor-intensive. In this work, we investigate the ability of MALDI-TOF-MS to characterize and discriminate between closely related environmental strains of Escherichia coli and to classify them according to their respective sources. We compared the performance of a rapid MALDI-TOF-MS-based method to a commonly used rep-PCR-based method that employs the BOX-A1R primer. Among the criteria evaluated were repeatability and the ability of each method to group E. coli isolates according to their respective sources. Our data suggest that the MALDI-TOF-MS-based approach has a lower repeatability level compared to rep-PCR but offers an improved ability to correctly assign E. coli isolates to specific source groups. In addition, we have identified five biomarkers that appear conserved among avian species. We conclude that MALDI-TOF-MS may represent a promising, novel and rapid approach to addressing the problem of fecal contamination of surface waters and warrants further investigation.

Influence of sampling depth on Escherichia coli concentrations in beach monitoring.

While the US Environmental Protection Agency's (EPA) Beaches Environmental Assessment and Coastal Health (BEACH) Act requires coastal and Great Lakes' states to implement plans for monitoring bacterial contamination of recreational beach water, exactly how this monitoring should occur has not been regulated. This study examined differences in concentration of Escherichia coli in water collected from different depths and from different horizontal locations across the beach. E. coli concentrations were significantly different (p<0.05), when water from different depths was compared. Sampling water at depths of 30, 60, and 120 cm resulted in significantly lower E. coli concentrations as depth increased. Had the State of Wisconsin chosen to collect beach water monitoring samples at a shallower or deeper depth, numbers of beach closures and the potential risk to public health would have changed substantially. These data imply that a revised and standardized protocol for monitoring beach water should be adopted by all states of a monitoring region to better compare microbial contamination of beaches and protect public health.

Effects of temperature and sand on E. coil survival in a northern lake water microcosm.

A concern for public health officials is the presence of Escherichia coli (E. coli), an indicator of fecal contamination, in monitoring recreational waters. While E. coli is unlikely to cause disease in humans, its presence may indicate other more pathogenic microorganisms. Many factors can lead to changes in the survival of E. coli outside of the animal intestine and may affect the probability of colonizing a new host. Survival of bacteria in recreational water has been linked to water temperature, and most recently to the presence of sand on the beach. This project looked at the survival of an environmental E. coli isolate in lake water. Lake water microcosms were placed at 4, 10, 14, or 25 degrees C for up to 36 d and an enzyme-substrate test (Colisure, IDEXX Corp.) was used to determine the most probable number (MPN) of E. coli/100 ml water. E. coli numbers at all temperatures declined over the duration of the experiment. The decline was most pronounced at 14 degrees C and was slowest at 4 degrees C. The presence of sand in the microcosm increased the time that E. coli survived, regardless of temperature. From a beach management standpoint, these findings indicate that E.coli may persist in the environment in cooler water longer than in the warmer water encountered in late summer.

Using a dopamine type 1A receptor agonist in high-risk patients to ameliorate contrast-associated nephropathy.

OBJECTIVE: The objective of our study was to evaluate the effects of fenoldopam mesylate, a dopamine type 1A receptor agonist and a potent renal vasodilator that markedly increases renal blood flow, on kidney function of patients who were receiving iodinated contrast material for an interventional procedure and thought to be at high risk of contrast-associated nephropathy. MATERIALS AND METHODS: We retrospectively reviewed the records of all patients who received fenoldopam mesylate to determine the acute and, when possible, the longer term effects on kidney function. RESULTS: Twenty-nine cases were reviewed. The average serum creatinine value before contrast administration was 2.55 mg/dL (range, 1.3-5.8 mg/dL) [corrected]. Twenty-four hours after contrast administration, serum creatinine was measured in 28 of the 29 patients. The serum creatinine values had decreased in 16 of the 28 patients by an average of 0.55 mg/dL [corrected]. In nine patients, the serum creatinine value had not changed. Two of the three increases in the serum creatinine value appear to have been caused primarily by problems that did not involve the contrast material. CONCLUSION: The use of fenoldopam mesylate at appropriate doses offers patients at high risk for contrast-associated nephropathy a chance to avoid this complication. To learn the extent and true nature of the effect of fenoldopam mesylate in this patient population requires a rigorous scientific trial, which is currently underway.