Kristen P. Brenner

Rapidly measured indicators of recreational water quality are predictive of swimming-associated gastrointestinal illness.

Standard methods to measure recreational water quality require at least 24 hr to obtain results, making it impossible to assess the quality of water within a single day. Methods to measure recreational water quality in ≤ 2 hr have been developed. Application of rapid methods could give considerably more accurate and timely assessments of recreational water quality. We conducted a prospective study of beachgoers at two Great Lakes beaches to examine the association between recreational water quality, obtained using rapid methods, and gastrointestinal (GI) illness after swimming. Beachgoers were asked about swimming and other beach activities and 10–12 days later were asked about the occurrence of GI symptoms. We tested water samples for Enterococcus and Bacteroides species using the quantitative polymerase chain reaction (PCR) method. We observed significant trends between increased GI illness and Enterococcus at the Lake Michigan beach and a positive trend for Enterococcus at the Lake Erie beach. The association remained significant for Enterococcus when the two beaches were combined. We observed a positive trend for Bacteroides at the Lake Erie beach, but no trend was observed at the Lake Michigan beach. Enterococcus samples collected at 0800 hr were predictive of GI illness that day. The association between Enterococcus and illness strengthened as time spent swimming in the water increased. This is the first study to show that water quality measured by rapid methods can predict swimming-associated health effects.

High sensitivity of children to swimming-associated gastrointestinal illness: results using a rapid assay of recreational water quality.

Background: Culture-based methods of monitoring fecal pollution in recreational waters require 24 to 48 hours to obtain results. This delay leads to potentially inaccurate management decisions regarding beach safety. We evaluated the quantitative polymerase chain reaction (QPCR) as a faster method to assess recreational water quality and predict swimming-associated illnesses. Methods: We enrolled visitors at 4 freshwater Great Lakes beaches, and contacted them 10 to 12 days later to ask about health symptoms experienced since the visit. Water at the beaches was polluted by point sources that carried treated sewage. We tested water samples daily for Enterococcus using QPCR and membrane filtration (EPA Method 1600). Results: We completed 21,015 interviews and tested 1359 water samples. Enterococcus QPCR cell equivalents (CEs) were positively associated with swimming-associated gastrointestinal (GI) illness (adjusted odds ratio per 1 log10 QPCR CE =1.26; 95% confidence interval = 1.06–1.51). The association between GI illness and QPCR CE was stronger among children aged 10 years and below (1.69; 1.24–2.30). Nonenteric illnesses were not consistently associated with Enterococcus QPCR CE exposure, although rash and earache occurred more frequently among swimmers. Enterococcus QPCR CE exposure was more strongly associated with GI illness than Enterococcus measured by membrane filtration. Conclusions: Measurement of the indicator bacteria Enterococci in recreational water using a rapid QPCR method predicted swimming-associated GI illness at freshwater beaches polluted by sewage discharge. Children at 10 years or younger were at greater risk for GI illness following exposure.

Rapidly measured indicators of recreational water quality and swimming-associated illness at marine beaches: a prospective cohort study.

IntroductionIn the United States and elsewhere, recreational water quality is monitored for fecal indicator bacteria to help prevent swimming-associated illnesses. Standard methods to measure these bacteria take at least 24 hours to obtain results. Molecular approaches such as quantitative polymerase chain reaction (qPCR) can estimate these bacteria faster, in under 3 hours. Previously, we demonstrated that measurements of the fecal indicator bacteria Enterococcus using qPCR were associated with gastrointestinal (GI) illness among swimmers at freshwater beaches. In this paper, we report on results from three marine beach sites.MethodsWe interviewed beach-goers and collected water samples at marine beaches affected by treated sewage discharges in Mississippi in 2005, and Rhode Island and Alabama in 2007. Ten to twelve days later, we obtained information about gastrointestinal, respiratory, eye, ear and skin symptoms by telephone. We tested water samples for fecal indicator organisms using qPCR and other methods.ResultsWe enrolled 6,350 beach-goers. The occurrence of GI illness among swimmers was associated with a log10-increase in exposure to qPCR-determined estimates of fecal indicator organisms in the genus Enterococcus (AOR = 2.6, 95% CI 1.3-5.1) and order Bacteroidales (AOR = 1.9, 95% CI 1.3-2.9). Estimates of organisms related to Clostridium perfringens and a subgroup of organisms in the genus Bacteroides were also determined by qPCR in 2007, as was F+ coliphage, but relationships between these indicators and illness were not statistically significant.ConclusionsThis study provides the first evidence of a relationship between gastrointestinal illness and estimates of fecal indicator organisms determined by qPCR at marine beaches.

Contact with beach sand among beachgoers and risk of illness.

Recent studies of beach sand fecal contamination have triggered interest among scientists and in the media. Although evidence shows that beach sand can harbor high concentrations of fecal indicator organisms, as well as fecal pathogens, illness risk associated with beach sand contact is not well understood. Beach visitors at 7 US beaches were enrolled in the National Epidemiological and Environmental Assessment of Recreational Water (NEEAR) Study during 2003-2005 and 2007 and asked about sand contact on the day of their visit to the beach (digging in the sand, body buried in the sand). Then, 10-12 days after their visit, participants were telephoned to answer questions about any health symptoms experienced since the visit. The authors completed 27,365 interviews. Digging in the sand was positively associated with gastrointestinal illness (adjusted incidence proportion ratio (aIPR) = 1.13, 95% confidence interval (CI): 1.02, 1.25) and diarrhea (aIPR = 1.20, 95% CI: 1.05, 1.36). The association was stronger between those buried in the sand and gastrointestinal illness (aIPR = 1.23, 95% CI: 1.05, 1.43) and diarrhea (aIPR = 1.24, 95% CI: 1.01, 1.52). Nonenteric illnesses did not show a consistent association with sand contact activities. Sand contact activities were associated with enteric illness at beach sites. Variation in beach-specific results suggests that site-specific factors may be important in the risk of illness following sand exposure.

Fecal indicators in sand, sand contact, and risk of enteric illness among beachgoers

Background: Beach sand can harbor fecal indicator organisms and pathogens, but enteric illness risk associated with sand contact remains unclear. Methods: In 2007, visitors at 2 recreational marine beaches were asked on the day of their visit about sand contact. Ten to 12 days later, participants answered questions about health symptoms since the visit. F+ coliphage, Enterococcus, Bacteroidales, fecal Bacteroides, and Clostridium spp. in wet sand were measured using culture and molecular methods. Results: We analyzed 144 wet sand samples and completed 4999 interviews. Adjusted odds ratios (aORs) were computed, comparing those in the highest tertile of fecal indicator exposure with those who reported no sand contact. Among those digging in sand compared with those not digging in sand, a molecular measure of Enterococcus spp. (calibrator cell equivalents/g) in sand was positively associated with gastrointestinal (GI) illness (aOR = 2.0 [95% confidence interval (CI) = 1.2–3.2]) and diarrhea (2.4 [1.4–4.2]). Among those buried in sand, point estimates were greater for GI illness (3.3 [1.3–7.9]) and diarrhea (4.9 [1.8–13]). Positive associations were also observed for culture-based Enterococcus (colony-forming units/g) with GI illness (aOR digging = 1.7 [1.1–2.7]) and diarrhea (2.1 [1.3–3.4]). Associations were not found among nonswimmers with sand exposure. Conclusions: We observed a positive relationship between sand-contact activities and enteric illness as a function of concentrations of fecal microbial pollution in beach sand.

Modeling Enterococcus densities measured by quantitative polymerase chain reaction and membrane filtration using environmental conditions at four Great Lakes beaches.

Data collected by the US Environmental Protection Agency (EPA) during the summer months of 2003 and 2004 at four US Great Lakes beaches were analyzed using linear regression analysis to identify relationships between meteorological, physical water characteristics, and beach characteristics data and the fecal indicator bacteria, Enterococcus. Water samples were analyzed for Enterococcus densities by quantitative polymerase chain reaction (qPCR) and membrane filtration (MF). This paper investigates the ability of regression models to accurately predict Enterococcus densities above or below a threshold value, using environmental data on a beach-by-beach basis for both methods. The ability to create statistical models for real-time water quality analysis would allow beach managers to make more accurate decisions regarding beach safety. Results from linear regression models indicate that environmental factors explain more of the variability in Enterococcus densities measured by MF than Enterococcus densities measured by qPCR. Results also show that models for both methods did not perform well at predicting occurrences in which water quality levels exceeded a threshold.

Water quality, weather and environmental factors associated with fecal indicator organism density in beach sand at two recreational marine beaches

Recent studies showing an association between fecal indicator organisms (FIOs) in sand and gastrointestinal (GI) illness among beachgoers with sand contact have important public health implications because of the large numbers of people who recreate at beaches and engage in sand contact activities. Yet, factors that influence fecal pollution in beach sand remain unclear. During the 2007 National Epidemiological and Environmental Assessment of Recreational (NEEAR) Water Study, sand samples were collected at three locations (60 m apart) on weekend days (Sat, Sun) and holidays between June and September at two marine beaches - Fairhope Beach, AL and Goddard Beach, RI - with nearby publicly-owned treatment works (POTWs) outfalls. F(+) coliphage, enterococci, Bacteroidales, fecal Bacteroides spp., and Clostridium spp. were measured in sand using culture and qPCR-based calibrator-cell equivalent methods. Water samples were also collected on the same days, times and transects as the 144 sand samples and were assayed using the same FIO measurements. Weather and environmental data were collected at the time of sample collection. Mean FIO concentrations in sand varied over time, but not space. Enterococci CFU and CCE densities in sand were not correlated, although other FIOs in sand were. The strongest correlation between FIO density in sand and water was fecal Bacteroides CCE, followed by enterococci CFU, Clostridium spp. CCE, and Bacteroidales CCE. Overall, the factors associated with FIO concentrations in sand were related to the sand-water interface (i.e., sand-wetting) and included daily average densities of FIOs in water, rainfall, and wave height. Targeted monitoring that focuses on daily trends of sand FIO variability, combined with information about specific water quality, weather, and environmental factors may inform beach monitoring and management decisions to reduce microbial burdens in beach sand. The views expressed in this paper are those of the authors and do not necessarily reflect the views or policies of the U.S. Environmental Protection Agency.

Standardization of enterococci density estimates by EPA qPCR methods and comparison of beach action value exceedances in river waters with culture methods.

The U.S. EPA has published recommendations for calibrator cell equivalent (CCE) densities of enterococci in recreational waters determined by a qPCR method in its 2012 recreational water quality criteria (RWQC). The CCE quantification unit stems from the calibration model used to estimate enterococci densities in recreational beach waters in the EPA National Epidemiological and Environmental Assessment of Recreational (NEEAR) Water Study and directly informed the derivation of the RWQC recommendations. Recent studies have demonstrated that CCE estimates from the method can vary when using different cultured Enterococcus cell preparations in calibrator samples. These differences have been attributed to differences in the quantities of targeted gene copies (target sequences) that are recovered per nominal calibrator cell by DNA extraction. Standardization of results from the calibration model will require the estimation of target sequence recoveries from the calibrator and water samples. In addition, comparisons of water sample results with the RWQC values will require a knowledge of target sequence recoveries from the NEEAR study calibrator samples. In this study recoveries of target sequences and the mean target sequence/cell ratio for the NEEAR study calibrator samples were retrospectively estimated with a corroborated standard curve. A modification of the calibration model was then used to estimate enterococci target sequence quantities in water samples from eight midwestern U.S. rivers. CCE estimates were obtained by dividing these target sequence quantities by the mean NEEAR study target sequence/cell ratio. This target sequence-based quantification approach resulted in a high degree of agreement in beach action decisions (determinations of whether bacterial fecal indicator densities are above or below RWQC-recommended values) from CCE results of the qPCR method and from culture dependent enumeration of both enterococci and Eschericia coli in the corresponding water samples.

Influence of wastewater disinfection on densities of culturable fecal indicator bacteria and genetic markers

The US Environmental Protection Agency has proposed the use of quantitative polymerase chain reaction (qPCR) as a rapid alternative analytical method for monitoring recreational water quality at beaches. For qPCR to be considered for other Clean Water Act purposes, such as inclusion in discharge permits and use in Total Maximum Daily Load calculations, it is necessary to understand how qPCR detectable genetic markers are influenced by wastewater disinfection. This study investigated genetic markers for Escherichia coli, Enterococcus, Clostridium spp., Bacteroides, total Bacteroidales, as well as the human-associated Bacteroides markers, HF183 and HumM2, to determine which, if any, were influenced by disinfection (chlorination or ultraviolet light) of effluents from secondary wastewater treatment in different seasons. The effects of disinfection on culturable enterococci, E. coli, Bacteroides, and C. perfringens were also compared to their associated genetic markers. Disinfection of secondary treatment effluents significantly reduced culturable fecal indicator bacteria (FIB) but not genetic marker densities. No significant differences were observed in the responses of FIB culture and genetic marker densities to type of disinfection (chlorination vs UV) or season. Results of this study provide evidence that qPCR may not be suitable for monitoring efficacy of wastewater disinfection on the inactivation of bacterial pathogens.

Dietary characterizations in a study of human exposures in the Lower Rio Grande Valley: II. Household waters

Abstract The Lower Rio Grande Valley Environmental Study (LRGVES) was designed to evaluate multiple forms of exposure to Valley residents because of community concerns of possible adverse health effects from environmental conditions. This is the second of two papers that describe the dietary components of the LRGVES scoping study, conducted in the spring and summer of 1993 in nine Valley residences. Drinking water and household water samples were collected along with food and beverage samples to determine potential dietary exposures of the participants. Water samples were analyzed for various organic compounds, including volatile organic compounds, polycyclic aromatic hydrocarbons, pesticides, haloacetic acids, chlorinated acids, and benzidines. Samples were also analyzed for trace elements, selected anions, and microbial quality. Sources of water available in the residences for drinking and other household purposes were public treatment facilities, vended water machines, and a private well. In general, organic and inorganic chemicals found in commercial sources of drinking water did not exceed federal criteria. However, products of the disinfection process (total trihalomethanes) exceeded the federal regulatory level in one sample. In addition, tap water samples from public water supplies were high in chloride and sulfate anions, often related to poor odor, color, or taste, resulting in residents of the Valley purchasing water for drinking at local vending machines and storing it in their homes in a variety of containers. Microbiological contamination of these containers was found to be a potential health problem. Coliform levels exceeding federal criteria for public water supplies were found in the water of several participants who did not regularly disinfect the containers in which they stored their drinking water. Water from a private well, which was designated for purposes other than drinking, was also unsuitable for drinking because of microbial contamination. Water sampling in the LRGVES pilot study identified the need for intervention and educational programs to eliminate exposures of Valley residents.