Research gaps and priorities for quantitative microbial risk assessment (QMRA).

The coronavirus disease 2019 pandemic highlighted the need for more rapid and routine application of modeling approaches such as quantitative microbial risk assessment (QMRA) for protecting public health. QMRA is a transdisciplinary science dedicated to understanding, predicting, and mitigating infectious disease risks. To better equip QMRA researchers to inform policy and public health management, an Advances in Research for QMRA workshop was held to synthesize a path forward for QMRA research. We summarize insights from 41 QMRA researchers and experts to clarify the role of QMRA in risk analysis by (1) identifying key research needs, (2) highlighting emerging applications of QMRA; and (3) describing data needs and key scientific efforts to improve the science of QMRA. Key identified research priorities included using molecular tools in QMRA, advancing dose-response methodology, addressing needed exposure assessments, harmonizing environmental monitoring for QMRA, unifying a divide between disease transmission and QMRA models, calibrating and/or validating QMRA models, modeling co-exposures and mixtures, and standardizing practices for incorporating variability and uncertainty throughout the source-to-outcome continuum. Cross-cutting needs identified were to: develop a community of research and practice, integrate QMRA with other scientific approaches, increase QMRA translation and impacts, build communication strategies, and encourage sustainable funding mechanisms. Ultimately, a vision for advancing the science of QMRA is outlined for informing national to global health assessments, controls, and policies.

Underestimation of Per- and Polyfluoroalkyl Substances in Biosolids: Precursor Transformation During Conventional Treatment.

Wastewater treatment plants generate a solid waste known as biosolids. The most common management option for biosolids is to beneficially reuse them as an agricultural amendment, but because of the risk of pathogen exposure, many regulatory bodies require pathogen reduction before biosolids reuse. Per- and polyfluoroalkyl substances (PFAS) are well documented in biosolids, but limited information is available on how biosolids treatment processes impact PFAS. Furthermore, quantification of PFAS has focused on perfluoroalkyl acids (PFAAs) which are a small fraction of thousands of PFAS known to exist. The objective of this study was to quantify 92 PFAS in biosolids collected from eight biosolids treatment facilities before and after four pathogen treatment applications: composting, heat treatment, lime treatment, and anaerobic digestion. Overall, total PFAS concentrations before and after treatment were dominated by PFAA precursor species, in particular, diPAPs which accounted for a majority of the mass of the Σ92PFAS. This differs from historic data that found PFAAs, primarily PFOS, to dominate total PFAS concentrations. Treatment options such as heat treatment and composting changed the ratio of PFAA precursors to PFAAs indicating a transformation of PFAS during treatment. This study finds that PFAA precursors are likely underrepresented by other studies and make up a larger percentage of the total PFAS concentration in biosolids than previously estimated.

Per- and Polyfluoroalkyl Substances (PFAS) in Street Sweepings.

One hundred and seventeen street sweeping samples were collected and analyzed for per- and polyfluoroalkyl substances (PFAS). Fifty-six samples were collected in one city (Gainesville, Florida) allowing for an in-depth city-wide characterization. Street sweepings from five other urban areas, (Orlando, n = 15; Key West, n = 15; Pensacola, n = 12; Tampa, n = 13; and Daytona Beach, n = 6) were analyzed to provide a city-to-city comparison of PFAS. Within our analytical workflow, 37 PFAS were quantified across all samples, while the maximum number of PFAS quantified at one site was 26. Of those PFAS quantified in Gainesville, 60% were perfluoroalkyl acids (PFAAs) and 33% were precursors to PFAA. Among the PFAAs, short-chain perfluoroalkyl carboxylic acids (PFCAs) were the dominant class representing 26% of the total PFAS by concentration. In the comparison across different urban cities, the dominant compound by concentration and frequency of detection varied; however, perfluorooctanoic acid (PFOA) and linear perfluorooctanesulfonic acid (PFOSlin) were the two PFAS that were detected the most frequently. This study documents the first-time detection of hexadecafluorosebacic acid and perfluoro-3,6,9-trioxaundecane-1,11-dioic acid within environmental samples.

Review of methods to determine hand surface area of children less than six years old: a case study.

Various methods exist to determine the surface area of hands. The consistency of these methods is essential given that risk assessments utilize hand surface area (HSA) to quantify exposure to environmental contaminants. HSA is also utilized in the clinical setting to estimate size of burns, and to determine specific treatments and medication dosages. A reliable method of surface area measurement is important to guide these decisions, especially in children who are vulnerable to environmental contaminants and medication side effects. Despite this, fewer HSA-determining studies have been performed for children compared to adults. In this study, 122 children completed hand tracings, and the tracings were digitized using an ImageJ program to determine HSA. Six previously published methods of determining HSA were utilized based on the child's height, weight, and length and width of hand. Children were analyzed by age group including 0-2, 3-4, and 5-6 years. The HSA measurements determined by five of the six methods were statistically different from HSA determined using direct hand tracings/Image J methodology (p < 0.001). The single remaining study that did not differ significantly from the hand tracing method provided a uniform hand to total body surface area (TBSA) ratio for children of all ages. Based on these results, we propose a novel age-group-specific ratio utilizing the HSA results from hand tracings and TBSA calculations. The percentages of TBSA that reflect HSA for children aged 0-2, 3-4 and 5-6 years were 0.91%, 0.90% and 0.87%, respectively. These percentages should be considered for use in risk assessments and the clinical setting to guide treatment and prognosis.

Towards integrated modeling of the long-term impacts of oil spills.

Although great progress has been made to advance the scientific understanding of oil spills, tools for integrated assessment modeling of the long-term impacts on ecosystems, socioeconomics and human health are lacking. The objective of this study was to develop a conceptual framework that could be used to answer stakeholder questions about oil spill impacts and to identify knowledge gaps and future integration priorities. The framework was initially separated into four knowledge domains (ocean environment, biological ecosystems, socioeconomics, and human health) whose interactions were explored by gathering stakeholder questions through public engagement, assimilating expert input about existing models, and consolidating information through a system dynamics approach. This synthesis resulted in a causal loop diagram from which the interconnectivity of the system could be visualized. Results of this analysis indicate that the system naturally separates into two tiers, ocean environment and biological ecosystems versus socioeconomics and human health. As a result, ocean environment and ecosystem models could be used to provide input to explore human health and socioeconomic variables in hypothetical scenarios. At decadal-plus time scales, the analysis emphasized that human domains influence the natural domains through changes in oil-spill related laws and regulations. Although data gaps were identified in all four model domains, the socioeconomics and human health domains are the least established. Considerable future work is needed to address research gaps and to create fully coupled quantitative integrative assessment models that can be used in strategic decision-making that will optimize recoveries from future large oil spills.

Possible impacts of sea level rise on disease transmission and potential adaptation strategies, a review.

Sea levels are projected to rise in response to climate change, causing the intrusion of sea water into land. In flat coastal regions, this would generate an increase in shallow water covered areas with limited circulation. This scenario raises a concern about the consequences it could have on human health, specifically the possible impacts on disease transmission. In this review paper we identified three categories of diseases which are associated with water and whose transmission can be affected by sea level rise. These categories include: mosquitoborne diseases, naturalized organisms (Vibrio spp. and toxic algae), and fecal-oral diseases. For each disease category, we propose comprehensive adaptation strategies that would help minimize possible health risks. Finally, the City of Key West, Florida is analyzed as a case study, due to its inherent vulnerability to sea level rise. Current and projected adaptation techniques are discussed as well as the integration of additional recommendations, focused on disease transmission control. Given that sea level rise will likely continue into the future, the promotion and implementation of positive adaptation strategies is necessary to ensure community resilience.

Effect of beach management policies on recreational water quality.

When beach water monitoring programs identify poor water quality, the causes are frequently unknown. We hypothesize that management policies play an important role in the frequency of fecal indicator bacteria (FIB) exceedances (enterococci and fecal coliform) at recreational beaches. To test this hypothesis we implemented an innovative approach utilizing large amounts of monitoring data (n > 150,000 measurements per FIB) to determine associations between the frequency of contaminant exceedances and beach management practices. The large FIB database was augmented with results from a survey designed to assess management policies for 316 beaches throughout the state of Florida. The FIB and survey data were analyzed using t-tests, ANOVA, factor analysis, and linear regression. Results show that beach geomorphology (beach type) was highly associated with exceedance of regulatory standards. Low enterococci exceedances were associated with open coast beaches (n = 211) that have sparse human densities, no homeless populations, low densities of dogs and birds, bird management policies, low densities of seaweed, beach renourishment, charge access fees, employ lifeguards, without nearby marinas, and those that manage storm water. Factor analysis and a linear regression confirmed beach type as the predominant factor with secondary influences from grooming activities (including seaweed densities and beach renourishment) and beach access (including charging fees, employing lifeguards, and without nearby marinas). Our results were observable primarily because of the very large public FIB database available for analyses; similar approaches can be adopted at other beaches. The findings of this research have important policy implications because the selected beach management practices that were associated with low levels of FIB can be implemented in other parts of the US and around the world to improve recreational beach water quality.

Using probabilities of enterococci exceedance and logistic regression to evaluate long term weekly beach monitoring data.

Recreational water quality surveillance involves comparing bacterial levels to set threshold values to determine beach closure. Bacterial levels can be predicted through models which are traditionally based upon multiple linear regression. The objective of this study was to evaluate exceedance probabilities, as opposed to bacterial levels, as an alternate method to express beach risk. Data were incorporated into a logistic regression for the purpose of identifying environmental parameters most closely correlated with exceedance probabilities. The analysis was based on 7,422 historical sample data points from the years 2000-2010 for 15 South Florida beach sample sites. Probability analyses showed which beaches in the dataset were most susceptible to exceedances. No yearly trends were observed nor were any relationships apparent with monthly rainfall or hurricanes. Results from logistic regression analyses found that among the environmental parameters evaluated, tide was most closely associated with exceedances, with exceedances 2.475 times more likely to occur at high tide compared to low tide. The logistic regression methodology proved useful for predicting future exceedances at a beach location in terms of probability and modeling water quality environmental parameters with dependence on a binary response. This methodology can be used by beach managers for allocating resources when sampling more than one beach.

Non-traditional abiotic drivers explain variability of chlorophyll-a in a shallow estuarine embayment.

Understanding the factors influencing eutrophication, as represented by concentrations of chlorophyll-a (Chl-a), is needed to inform effective management and conservation strategies promoting ecological resilience. The objective of this study was to evaluate a unique combination of abiotic explanatory factors to describe Chl-a concentrations within the study estuary (North Biscayne Bay, Florida, USA). Multiple linear regression determined the strength and direction of influence of factors using data from 10 water quality monitoring stations. The analysis also considered time scales for evaluating cumulative effects of freshwater inflow and wind. Results show that dominant drivers of Chl-a were temperature, freshwater volume (whose cumulative effects were evaluated up to a 60-day time scale), and turbidity, which were statistically significant at 60, 60, and 70 % of the investigated stations, respectively. All drivers collectively accounted for 22 to 63 % of the variability of Chl-a measurements. Of the nine variables evaluated, nutrient concentrations (orthophosphate and ammonia) were not among the top three overall drivers. Despite nutrients historically being cited in the literature as the most significant factor, this study asserts that non-nutrient factors often govern Chl-a levels, necessitating a paradigm shift in management strategies to bolster estuarine resilience against climate change.

Evaluation of per- and polyfluoroalkyl substances (PFAS) released from two Florida landfills based on mass balance analyses.

Per- and polyfluoroalkyl substances (PFAS) have been found at high levels within landfill environments. To assess PFAS distributions, this study aimed to evaluate PFAS mass flux leached from disposed solid waste and within landfill reservoirs by mass balance analyses for two full-scale operational Florida landfills. PFAS mass flux in different aqueous components within landfills were estimated based on PFAS concentrations and water flow rates. For PFAS concentration, 26 PFAS, including 18 perfluoroalkyl acids (PFAAs) and 8 PFAA-precursors, were measured in samples collected from the landfills or estimated based on previous studies. Flow rates of aqueous components (rainfall, evapotranspiration, runoff, stormwater, groundwater, leakage, gas condensate, and leachate) were evaluated through the Hydrologic Evaluation of Landfill Performance model, water balance, and Darcy's Law. Results showed that the average PFAS mass flux leached from the solid waste standardized by area was estimated as 36.8 g/ha-yr, which was approximately 1 % to 3 % of the total amount of PFAS within the solid waste. The majority of PFAS leached from the solid waste (95 % to 97 %) is captured by the leachate collection system, with other aqueous components representing much smaller fractions (stormwater system at 3 % to 5 %, and gas condensate and groundwater at < 1 %). Also, based on the results, we estimate that PFAS releases will likely occur at least over 40 years. Overall, these results can help prioritize components for waste management and PFAS treatment during the anticipated landfill release periods.

Comparison of the PFAS and physical-chemical parameter fluctuations between an ash landfill and a MSW landfill.

Studies of per- and polyfluoroalkyl substances (PFAS) fluctuations at landfills have focused on municipal solid waste (MSW) leachate. Few studies exist that evaluate fluctuations (defined by the coefficient of variation, CV) in MSW incinerator ash (MSWA) landfill leachate and that evaluate PFAS fluctuations in stormwater, groundwater, and treated liquids on-site. In this study, aqueous landfill samples (leachate, treated leachate, stormwater, gas condensate, ambient groundwater, and effluent from a groundwater remediation system) were collected from a MSW and an MSWA landfill geographically located within close proximity (less than 40 km). The objective of this study was to compare the leachate compositions between these two landfill types and to evaluate temporal variations. Results indicated that the CV of total detected PFAS concentrations in leachate was higher for the MSW landfill (CV = 43 %) compared to the MSWA landfill (CV = 16 %). The total detected PFAS concentration in MSW leachate samples (mean: 9641 ng/L) was higher than in MSWA leachate samples (mean: 2621 ng/L) (p < 0.05). Within a landfill, PFAS concentrations were correlated (rs > 0.6, p < 0.05) with alkalinity, total organic carbon (TOC), and ammonia. Results from the on-site leachate treatment system at the MSW landfill indicated reductions in COD, TOC, and ammonia; however, the ∑26PFAS concentration increased 3 % after the treatment. Overall, results demonstrated that differences between landfill types and fluctuations in PFAS within landfills should be considered when designing landfill leachate collection and treatment systems to remove PFAS. The comparative analysis in this study can provide insights into optimizing leachate management for MSW and MSWA landfills.

Evaluation of per- and polyfluoroalkyl substances (PFAS) in landfill liquids from Pennsylvania, Colorado, and Wisconsin.

PER: and polyfluoroalkyl substances (PFAS) have been measured in aqueous components within landfills. To date, the majority of these studies have been conducted in Florida. This current study aimed to evaluate PFAS concentrations in aqueous components (leachate, gas condensate, stormwater, and groundwater) from four landfills located outside of Florida, in Pennsylvania, Colorado, and Wisconsin (2 landfills). The Pennsylvania landfill also provided the opportunity to assess a leachate treatment system. Sample analyses were consistent across studies including the measurements of 26 PFAS and physical-chemical parameters. For the four target landfills, average PFAS concentrations were 6,900, 22,000, 280, and 260 ng L-1 in the leachate, gas condensate, stormwater, and groundwater, respectively. These results were not significantly different than those observed for landfills in Florida except for the significantly higher PFAS concentrations in gas condensate compared to leachate. For on-site treatment at the Pennsylvania landfill, results suggest that the membrane biological bioreactor (MBBR) system performed similarly as aeration-based leachate treatment systems at Florida landfills resulting in no significant decreases in ∑26PFAS. Overall, results suggest a general consistency across US regions in PFAS concentrations within different landfill liquid types, with the few differences observed likely influenced by landfill design and local climate. Results confirm that leachate exposed to open air (e.g., in trenches or in treatment systems) have lower proportions of perfluoroalkyl acid precursors relative to leachate collected in enclosed pipe systems. Results also confirm that landfills without bottom liner systems may have relatively higher PFAS levels in adjacent groundwater and that landfills in wetter climates tend to have higher PFAS concentrations in leachate.

Characterization of per- and polyfluoroalkyl substances (PFAS) and other constituents in MSW landfill leachate from Puerto Rico.

Elevated per- and polyfluoroalkyl substance (PFAS) concentrations have been reported in municipal solid waste (MSW) landfill leachate with higher levels in wet and warmer subtropical climates. Information about landfill leachate characteristics is much more limited in tropical climates. In this study, 20 landfill leachate samples were collected from three MSW landfills on the tropical island of Puerto Rico and results were compared against landfills nationally and within Florida, USA. The samples collected in Puerto Rico underwent physical-chemical analysis, as well as a quantitative analysis of 92 PFAS. Samples described in this study include discrete leachate types, such as leachate, gas condensate, and leachate which has undergone on-site treatment (e.g., RO treatment, phytoremediation, lagoons). A total of 51 PFAS were detected above quantitation limits, including perfluorohexylphosphonic acid, a perfluoroalkyl acid (PFAA) which has not been reported previously in landfill leachate. ∑PFAS concentrations in this study (mean: 38,000 ng L-1), as well as concentrations of individual PFAS, are significantly higher than other reported MSW landfill leachate concentrations. The profiles of leachates collected from on-site treatment systems indicate possible transformation of precursor PFAS as a result of treatment processes - oxidizing conditions, for example, may facilitate aerobic transformation, increase the concentrations of PFAAs, and possibly increase the apparent ∑PFAS concentration. Extreme climate events, including rising temperatures and more frequent hurricanes, have placed additional strain on the solid waste management infrastructure on the island - adding complexity to an already challenging PFAS management issue. As concern grows over PFAS contamination in drinking water, these findings should inform solid waste and leachate management decisions in order to minimize PFAS emissions in island environments.

Correction: Ferguson et al. Soil, Hand, and Body Adherence Measures across Four Beach Areas: Potential Influence on Exposure to Oil Spill Chemicals. Int. J. Environ. Res. Public Health 2020, 17, 4196.

In the original publication [...].

Methodologies for the collection of parameters to estimate dust/soil ingestion for young children.

Background: Heavy metals, pesticides and a host of contaminants found in dust and soil pose a health risk to young children through ingestion. Dust/soil ingestion rates for young children can be estimated using micro-level activity time series (MLATS) as model inputs. MLATS allow for the generation of frequency and duration of children's contact activities, along with sequential contact patterns. Models using MLATS consider contact types, and transfer dynamics to assign mechanisms of contact and appropriate exposure factors for cumulative estimates of ingestion rates. Objective: The objective of this study is to describe field implementation, data needs, advanced field collection, laboratory methodologies, and challenges for integrating into and updating a previously validated physical-stochastic MLATS-based model framework called the Child-Specific Aggregate Cumulative Human Exposure and Dose (CACHED) model. The manuscript focuses on describing the methods implemented in the current study. Methods: This current multidisciplinary study (Dust Ingestion childRen sTudy [DIRT]) was implemented across three US regions: Tucson, Arizona; Miami, Florida and Greensboro, North Carolina. Four hundred and fifty participants were recruited between August 2021 to June 2023 to complete a 4-part household survey, of which 100 also participated in a field study. Discussion: The field study focused on videotaping children's natural play using advanced unattended 360° cameras mounted for participants' tracking and ultimately conversion to MLATS. Additionally, children's hand rinses were collected before and after recording, along with indoor dust and outdoor soil, followed by advanced mass analysis. The gathered data will be used to quantify dust/soil ingestion by region, sociodemographic variables, age groups (from 6 months to 6 years), and other variables for indoor/outdoor settings within an adapted version of the CACHED model framework. Significance: New innovative approaches for the estimation of dust/soil ingestion rates can potentially improve modeling and quantification of children's risks to contaminants from dust exposure.

Investigating the partitioning behavior of per- and polyfluoroalkyl substances (PFAS) during thermal landfill leachate evaporation.

Thermal landfill leachate evaporator systems can reduce the volume of leachate by up to 97%, while releasing water vapor and producing residuals (volume-reduced leachate and sludge) that are managed on-site. On-site thermal evaporators offer landfill operators leachate management autonomy without being subject to increasingly stringent wastewater treatment plant requirements. However, little is known about the partitioning of PFAS within these systems, nor the extent to which PFAS may be emitted into the environment via vapor. In this study, feed leachate, residual evaporated leachate, sludge, and condensed vapor were sampled at two active full-scale thermal landfill leachate evaporators and from a laboratory-scale leachate evaporation experiment. Samples were analyzed for 91 PFAS via ultra-high pressure liquid chromatography - tandem mass spectrometry (UHPLC-MS/MS). Similar trends were observed from Evaporator 1, Evaporator 2, and the laboratory-scale evaporator; ∑PFAS were concentrated in the residual evaporated leachate during evaporation by a factor of 5.3 to 20. All condensed vapors sampled (n = 5) contained PFAS, predominantly 5:3 fluorotelomer carboxylic acid (5:3FTCA), (full-scale vapors 729 - 4087 ng/L PFAS; lab-scale vapor 61.0 ng/L PFAS). For Evaporators 1 and 2, an estimated 9 - 24% and 10%, respectively, of the PFAS mass entering the evaporators in leachate was released with vapor during the days of sample collection. '.

Enterococci pathways to coastal waters and implications of sea level rise.

Highly urban coastal communities in low lying areas and with high water tables are vulnerable to sea-level rise and to corresponding increases in coastal groundwater levels. Stormwater conveyance systems are under increased risk. Rising groundwater levels affect the hydraulics of the stormwater system thereby increasing contaminant transport, for example the fecal indicator bacteria enterococci, to coastal waters. This study offers a unique opportunity to evaluate the impacts of increased contaminant transport on marine coastal environments. Here we assessed historic and recent coastal water quality, stormwater sampling data, groundwater monitoring and tidal elevations near the coastline, in the context of altered hydraulics within the system. Two pathways of enterococci to marine waters were identified. Direct discharge of contaminated stormwater runoff via the stormwater outfalls and tidally driven contaminated groundwater discharge. As sea level continues to rise, we hypothesize that a diminished unsaturated zone coupled with altered hydraulic conditions at the coastal groundwater zone will facilitate the transport of enterococci from urban sediments to the study site (Park View Waterway in Miami Beach, FL USA). We recommend improvements to the stormwater conveyance system, and maintenance of the sanitary sewer system to mitigate these impacts and minimize transport of enterococci, and other stormwater pollutants to coastal waters. The results of this study can be useful to interpret high enterococci levels in low lying coastal areas where groundwater is influenced by rising sea water levels.

Wastewater based surveillance can be used to reduce clinical testing intensity on a university campus.

Clinical testing has been a vital part of the response to and suppression of the COVID-19 pandemic; however, testing imposes significant burdens on a population. College students had to contend with clinical testing while simultaneously dealing with health risks and the academic pressures brought on by quarantines, changes to virtual platforms, and other disruptions to daily life. The objective of this study was to analyze whether wastewater surveillance can be used to decrease the intensity of clinical testing while maintaining reliable measurements of diseases incidence on campus. Twelve months of human health and wastewater surveillance data for eight residential buildings on a university campus were analyzed to establish how SARS-CoV-2 levels in the wastewater can be used to minimize clinical testing burden on students. Wastewater SARS-CoV-2 levels were used to create multiple scenarios, each with differing levels of testing intensity, which were compared to the actual testing volumes implemented by the university. We found that scenarios in which testing intensity fluctuations matched rise and falls in SARS-CoV-2 wastewater levels had stronger correlations between SARS-CoV-2 levels and recorded clinical positives. In addition to stronger correlations, most scenarios resulted in overall fewer weekly clinical tests performed. We suggest the use of wastewater surveillance to guide COVID-19 testing as it can significantly increase the efficacy of COVID-19 surveillance while reducing the burden placed on college students during a pandemic. Future efforts should be made to integrate wastewater surveillance into clinical testing strategies implemented on college campuses.

Human-associated methicillin-resistant Staphylococcus aureus from a subtropical recreational marine beach.

Reports of Staphylococcus aureus including methicillin-resistant S. aureus (MRSA) detected in marine environments have occurred since the early 1990 s. This investigation sought to isolate and characterize S. aureus from marine waters and sand at a subtropical recreational beach, with and without bathers present, in order to investigate possible sources and to identify the risks to bathers of exposure to these organisms. During 40 days over 17 months, 1,001 water and 36 intertidal sand samples were collected by either bathers or investigators at a subtropical recreational beach. Methicillin-sensitive S. aureus (MSSA) and MRSA were isolated and identified using selective growth media and an organism-specific molecular marker. Antimicrobial susceptibility, staphylococcal cassette chromosome mec (SCCmec) type, pulsed-field gel electrophoresis (PFGE) pattern, multi-locus sequence type (MLST), and staphylococcal protein A (spa) type were characterized for all MRSA. S. aureus was isolated from 248 (37 %) bather nearby water samples at a concentration range of <2-780 colony forming units per ml, 102 (31 %) ambient water samples at a concentration range of <2-260 colony forming units per ml, and 9 (25 %) sand samples. Within the sand environment, S. aureus was isolated more often from above the intertidal zone than from intermittently wet or inundated sand. A total of 1334 MSSA were isolated from 37 sampling days and 22 MRSA were isolated from ten sampling days. Seventeen of the 22 MRSA were identified by PFGE as the community-associated MRSA USA300. MRSA isolates were all SCCmec type IVa, encompassed five spa types (t008, t064, t622, t688, and t723), two MLST types (ST8 and ST5), and 21 of 22 isolates carried the genes for Panton-Valentine leukocidin. There was a correlation (r = 0.45; p = 0.05) between the daily average number of bathers and S. aureus in the water; however, no association between exposure to S. aureus in these waters and reported illness was found. This report supports the concept that humans are a potential direct source for S. aureus in marine waters.

Integrating microbial source tracking with quantitative microbial risk assessment to evaluate site specific risk based thresholds at two South Florida beaches.

Quantitative microbial risk assessment (QMRA) can be used to evaluate health risks associated with recreational beach use. This study developed a site-specific risk assessment using a novel approach that combined quantitative PCR-based measurement of microbial source tracking (MST) genetic markers (human, dog, and gull fecal bacteria) with a QMRA analysis of potential pathogen risk. Water samples (n = 24) from two recreational beaches were collected and analyzed for MST markers as part of a broader Beach Exposure And Child Health Study that examined child behavior interactions with the beach environment. We report here the measurements of fecal bacteria MST markers in the environmental DNA extracts of those samples and a QMRA analysis of potential health risks utilizing the results from the MST measurements in the water samples. Human-specific Bacteroides was enumerated by the HF183 Taqman qPCR assay, gull-specific Catellicoccus was enumerated by the Gull2 qPCR assay, and dog-specific Bacteroides was enumerated by the DogBact qPCR assay. Derived reference pathogen doses, calculated from the MST marker concentrations detected in recreational waters, were used to estimate the risk of gastrointestinal illness for both children and adults. Dose-response equations were used to estimate the probability of the risk of infection (Pinf) per a swimming exposure event. Based on the QMRA simulations presented in this study, the GI risk from swimming or playing in water containing a mixture of human and non-human fecal sources appear to be primarily driven by the human fecal source. However, the estimated median GI health risk for both beaches never exceeded the U.S. EPA risk threshold of 32 illnesses per 1,000 recreation events. Our research suggests that utilizing QMRA together with MST can further extend our understanding of potential recreational bather risk by identifying the source contributing the greatest risk in a particular location, therefore informing beach management responses and decision-making.