Bacteriophage as an Alternative Indicator for Microbiological Pollution at Marine Beaches

Beaches contaminated by waterborne pathogens cause a variety of diseases. Studies show enterococci, a common fecal indicator bacteria, are not associated with emerging pathogens and bacteriophages could be better indicators for waterborne pathogens. This study examines the occurrence of two bacteriophages and their relation with enterococci in a marine beach known for high levels of pollution. Monthly samples were collected from four sites at Saint Andrews and Clam Creek Beach on Jekyll Island. Tidal influences were investigated by collecting samples at 7 a.m., 12 p.m., 3 p.m., and 7 p.m. Temperature, salinity, dissolved oxygen, and pH were also collected. US EPA Method 1602 was used to enumerate somatic and F bacteriophage and Method 1600 for Enterococci. Enterococci concentrations ranged from \u3c1 to 60 CFU/ 100ml with lowest concentrations detected at 12 p.m. and highest at 4 p.m. Somatic bacteriophage numbers fluctuated between 9 and 30 PFU /100ml and F phage 1 to 4 PFU /100ml. For bacteriophages, highest concentrations were found at 7 a.m. There was no significant relationship between enterococci and bacteriophages. A significant correlation between somatic and F specific coliphage (p\u3c0.01), somatic phage and temperature (p\u3c0.027), and F phage and salinity (p\u3c0.05) was detected. Highest concentrations of bacteria and bacteriophages were detected during ebb tide, due to increasing inflow from creeks near each beach. No significant relationship between enterococci and viruses was detected. Correlations between environmental factors and phages indicate better representation of environmental influence on beach water quality such as freshwater inflow due to runoff. Further studies may show the relation between viruses, including Adenovirus and Norovirus, and potential sources of pollution. Monitoring environmental conditions provides better information on use of bacteriophages as alternative indicators for microbial pollution in recreational marine waters

Coliphage as an Indicator of the Quality of Beach Water to Protect the Health of Swimmers in Coastal Georgia

Background: Gastrointestinal disease affects millions of people in the United States and places a substantial economic burden upon healthcare systems. Recreational waters polluted with fecal material are a main source for transmission of gastrointestinal disease. Georgia beaches are monitored for the presence of fecal indicator bacteria, but these bacteria are not well associated with enteric viruses. The United States Environmental Protection Agency (US EPA) has recently proposed coliphage (a virus of Escherichia coli) as an alternative indicator of fecal contamination in recreational waters. The present study compares fecal indicator bacteria and coliphage concentrations at two Georgia beaches with adjacent creeks that have a history of pollution. Methods: For one year, samples and environmental data were collected from four sites on Jekyll Island, GA, during the peak swimming season and the off-season. Samples were processed using US EPA-approved methods for membrane filtration and plaque formation. Statistical analyses were performed using t-tests and Spearman correlations. Results: The highest numbers of enterococci and significant differences with coliphage were found at Saint Andrews Creek during the swimming season and the off-season. The enterococci concentrations at Clam Creek sites did not exceed recommended recreational water criteria. During the off-season, concentrations of enterococci and coliphages were different at Clam Creek sites, indicating a potential risk for presence of enteric virus when enterococci could not be detected. Conclusions: The US EPA has proposed to adapt coliphage concentrations as an alternative indicator of water pollution for routine beach monitoring nationally. The present study provides a background for adoption of this method in Georgia. Measures of enterococci do not provide sufficient information about the associated human health risk. Inclusion of these viral indicators will improve decision making for beach closures and for protection of the health of swimmers. Keywords: coliphage, enterococci, enteric viruses, beach water quality, health risk, Georgi

The EU Center of Excellence for Exascale in Solid Earth (ChEESE): Implementation, results, and roadmap for the second phase

The EU Center of Excellence for Exascale in Solid Earth (ChEESE) develops exascale transition capabilities in the domain of Solid Earth, an area of geophysics rich in computational challenges embracing different approaches to exascale (capability, capacity, and urgent computing). The first implementation phase of the project (ChEESE-1P; 2018¿2022) addressed scientific and technical computational challenges in seismology, tsunami science, volcanology, and magnetohydrodynamics, in order to understand the phenomena, anticipate the impact of natural disasters, and contribute to risk management. The project initiated the optimisation of 10 community flagship codes for the upcoming exascale systems and implemented 12 Pilot Demonstrators that combine the flagship codes with dedicated workflows in order to address the underlying capability and capacity computational challenges. Pilot Demonstrators reaching more mature Technology Readiness Levels (TRLs) were further enabled in operational service environments on critical aspects of geohazards such as long-term and short-term probabilistic hazard assessment, urgent computing, and early warning and probabilistic forecasting. Partnership and service co-design with members of the project Industry and User Board (IUB) leveraged the uptake of results across multiple research institutions, academia, industry, and public governance bodies (e.g. civil protection agencies). This article summarises the implementation strategy and the results from ChEESE-1P, outlining also the underpinning concepts and the roadmap for the on-going second project implementation phase (ChEESE-2P; 2023¿2026).This work has been funded by the European Union Horizon 2020 research and innovation program under the ChEESE project, Grant Agreemen

The EU Center of Excellence for Exascale in Solid Earth (ChEESE): Implementation, results, and roadmap for the second phase

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Global patient outcomes after elective surgery: prospective cohort study in 27 low-, middle- and high-income countries.

BACKGROUND: As global initiatives increase patient access to surgical treatments, there remains a need to understand the adverse effects of surgery and define appropriate levels of perioperative care. METHODS: We designed a prospective international 7-day cohort study of outcomes following elective adult inpatient surgery in 27 countries. The primary outcome was in-hospital complications. Secondary outcomes were death following a complication (failure to rescue) and death in hospital. Process measures were admission to critical care immediately after surgery or to treat a complication and duration of hospital stay. A single definition of critical care was used for all countries. RESULTS: A total of 474 hospitals in 19 high-, 7 middle- and 1 low-income country were included in the primary analysis. Data included 44 814 patients with a median hospital stay of 4 (range 2-7) days. A total of 7508 patients (16.8%) developed one or more postoperative complication and 207 died (0.5%). The overall mortality among patients who developed complications was 2.8%. Mortality following complications ranged from 2.4% for pulmonary embolism to 43.9% for cardiac arrest. A total of 4360 (9.7%) patients were admitted to a critical care unit as routine immediately after surgery, of whom 2198 (50.4%) developed a complication, with 105 (2.4%) deaths. A total of 1233 patients (16.4%) were admitted to a critical care unit to treat complications, with 119 (9.7%) deaths. Despite lower baseline risk, outcomes were similar in low- and middle-income compared with high-income countries. CONCLUSIONS: Poor patient outcomes are common after inpatient surgery. Global initiatives to increase access to surgical treatments should also address the need for safe perioperative care. STUDY REGISTRATION: ISRCTN5181700

Assessing the Human-health Risk of Exposure to Pathogens from Beach Sands

Fecal contamination at recreational beaches impacts the health of beachgoers, through the introduction of disease-causing microorganisms, and the well-being of communities dependent on income from recreational beach activities. Beach ecosystems are also impacted by sewage through the introduction of nutrients that can cause abnormal increases in autochthonous microorganisms which can impact the population of larger organisms in the ecosystem. Fecal contamination is introduced into sand via untreated sewage, direct deposition of human feces into sand, runoff, and deposition of animal feces into sand. The introduction of fecal contamination into sand exposes individuals to pathogens (disease causing microorganisms) which can result in gastrointestinal illness. While standard methods and regulatory guidelines exist for the monitoring of fecal contamination in water, none exist for sand despite the data that link recreational contact with sand to gastrointestinal illness. The detection of fecal-associated pathogens in the environment is difficult due to their diversity and low concentration, therefore contamination is monitored using fecal indicator bacteria (FIB). FIB are present in the gastrointestinal tract of animals and provide an indication of the presence of fecal contamination. Escherichia coli (freshwater) and enterococci (fresh and salt water) are commonly used as FIB globally and in the United States. In sand, FIB are present at higher concentrations than in water, as sand protects FIB from environmental stressors such as UV or predation and provides easier access to nutrients. FIB that survive long-term and replicate in sand are termed “naturalized.” Naturalized populations can complicate the identification of recent fecal contamination, as FIB monitoring techniques cannot differentiate between naturalized FIB and those recently introduced through sources of fecal contamination. The ubiquity of FIB among animals and humans means it is also difficult to determine the source of contamination. Some sources of fecal contamination (human as compared to cow or bird fecal contamination) contain a higher number of pathogens that are likely to infect humans (adenovirus in humans or Cryptosporidium spp. from cows). Therefore, microbial source tracking (MST) was developed to include a suite of host-associated genes (markers) of fecal microorganisms that are specific to different animals. These markers can differentiate between sources of fecal contamination, supplementing FIB monitoring and providing a more accurate depiction of the fecal contamination picture. Few studies have investigated the presence of MST markers in sand or determined relationships between MST markers and FIB in sand. The human health risk associated with exposure to fecal contamination can be assessed by epidemiological studies; however, these studies are typically expensive and require specialized teams that may not be available to organizations with limited resources. Quantitative microbial risk assessment (QMRA) is a mathematical modeling framework used to estimate human health risk (the likelihood someone becomes ill) from exposure to pathogens under different environmental scenarios. QMRA consists of four steps: Problem formulation, exposure assessment, dose-response modeling, and risk characterization. Problem formulation involves establishing the framework (reference pathogen identification, exposure pathway, sources of contamination) that helps target the risk management needs to be addressed. The exposure assessment then determines the concentration and frequency of exposure by individuals to the reference pathogen(s) and exposure pathway identified during the problem formulation step, also known as a dose. The dose is then compared to a dose-response model (typically sourced from the literature) to determine the probability that the estimated dose would lead to illness in an individual. Risk characterization then quantifies the level of risk based on the modeled data. Reverse QMRA, used in this experiment, estimates the probabilities of pathogen concentrations that correspond with a risk threshold that is defined by stakeholders. Few QMRAs have been conducted in sand, therefore, this dissertation is focused on understanding the human health risk from exposure to pathogens at a tropical beach impacted by sewage. Jacó beach is a tropical beach located on the Pacific coast of Costa Rica and is a popular tourist destination. A water quality study determined the fecal contamination was severely impacting the water quality of the beach and placing individuals at risk of illness from exposure to contaminated waters. Understanding the human health risk from exposure to pathogens in beach sand, in addition to the human health risk from water, ultimately helps to improve beach management decisions and public health. In chapter one, the analytical sensitivity of quantitative polymerase chain reaction (qPCR) for Enterococcus in sand was compared for the slurry (suspension, agitation, membrane filtration of supernatant), versus two direct extraction methods using PowerSoil™ or PowerMax Soil™ kits at a freshwater and saltwater beach in Tampa, Fl, USA. We found the slurry method had the lowest limit of detection at 20–80 gene copies g-1 (wet weight), recovered significantly more DNA, and was the only method that detected Enterococcus by qPCR in all samples; therefore, the slurry method was exclusively used in subsequent experiments. The slurry method reflected the spatial variability of Enterococcus in individual transect samples. Mean recovery efficiency of the human-associated microbial source tracking marker HF183 from marine and freshwater beach sand spiked with wastewater was 100.8% and 64.1%, respectively, but varied between dilutions, indicating that the mixing protocol needs improvement. The objective of chapter two was to determine the extent of the influence a contaminated waterbody has on the concentration of microbes in beach sand. Upstream, downstream and ocean samples were collected on a transect in sand at 0 m (origin), 2.5 m and 5.0 m from the riverbank or swash zone. Samples were also collected in sediment. Samples were analyzed for the presence of the MST marker HF183 and FIB Enterococcus using the slurry method followed by DNA extraction and qPCR. Median concentrations of Enterococcus decreased as distance from the river or ocean increased. Enterococcus ranged from 1.46 x 104 to 8.11 x 103 gene copies 100 g-1 to the same distance. HF183 and Enterococcus were positively correlated at the riverbank/swash zone, but not at other subsites due to frequent failure to detect HF183. Sediment samples did not differ in HF183 or Enterococcus concentrations along the river and HF183 was not detected in the ocean sediment. The frequency of detection for HF183 was significantly greater among samples along the riverbank than samples collected at the 2.5 m and 5 m subsites. Polluted waterbodies can influence microbe concentrations in sand but the extent of the influence of a waterbody on MST markers in sand requires further study. Chapter three employed a reverse QMRA in Copey River at Jacó Beach to estimate the varying probability that corresponds with meeting a risk target. The risk target was defined as 36 cases of gastrointestinal illness per 1000 people (36/1000), the health target set in the 2012 recreational water quality guidelines for the United States Environmental Protection Agency (USEPA). Data were used to calculate whether concentrations of Salmonella, Adenovirus, and Giardia exceeded or were below concentrations needed to meet a risk of 36/1000. Samples at two sites, one upstream and one downstream of where beachgoers tended to recreate at the riverbank were collected six times over three weeks. Samples were analyzed for the presence of enterococci (FIB), the human-host associated MST marker HF183 and sewage-associated MST marker PMMoV, and pathogens Salmonella, adenovirus, and Giardia. Sand samples were collected at the riverbank and analyzed for the fecal indicator bacteria enterococci, the human-host associated microbial source tracking marker genes HF183, sewage-associated marker pepper mild mottle virus (PMMoV), and pathogens Salmonella, Giardia, and human adenovirus. Enterococci and Salmonella were cultured using the slurry method followed by USEPA standard methods. HF183, PMMoV, and adenovirus were detected by (RT)-qPCR. Giardia was detected using microscopy following USEPA standard methodology. Enterococci were detected in all samples and had a geometric mean of 3.25 x 103 GC 100 g-1. Geometric means of the MST markers HF183 and PMMoV were 1.19 x 103 and 1.64 x103 GC 100 g-1 and were detected in 83% and 33% of samples, respectively. Salmonella was detected in 66.6% of samples and had a geometric mean of 2.45 x 102 CFU 100 g-1. Giardia and adenovirus were detected only once each at 45 oocysts 100 g-1 and 1.91 x 102 GC 100 g-1. QMRA analysis showed that adenovirus and Giardia concentrations exceeded the levels that coincide with the risk threshold of 36/1000 while Salmonella fell between the 50th and 75th % percentile probability of meeting the risk threshold. Exposure to sewage-borne pathogens in sand could contribute to the risk of illness for recreational users of Jacó Beach and should be analyzed in tandem with pathogens in water to implement the best strategy for the protection of public health. To implement a comprehensive beach monitoring strategy, it is important to understand all the contextual factors that affect public health at a beach. Recreation in sand has been linked to gastrointestinal illness, however there are no regulatory guidelines or standard methods for analysis of fecal microorganisms in sand. This research has shown that a comprehensive beach monitoring strategy should include sand monitoring strategies in tandem with water monitoring strategies

Assessing the Human-health Risk of Exposure to Pathogens from Beach Sands

Fecal contamination at recreational beaches impacts the health of beachgoers, through the introduction of disease-causing microorganisms, and the well-being of communities dependent on income from recreational beach activities. Beach ecosystems are also impacted by sewage through the introduction of nutrients that can cause abnormal increases in autochthonous microorganisms which can impact the population of larger organisms in the ecosystem. Fecal contamination is introduced into sand via untreated sewage, direct deposition of human feces into sand, runoff, and deposition of animal feces into sand. The introduction of fecal contamination into sand exposes individuals to pathogens (disease causing microorganisms) which can result in gastrointestinal illness. While standard methods and regulatory guidelines exist for the monitoring of fecal contamination in water, none exist for sand despite the data that link recreational contact with sand to gastrointestinal illness. The detection of fecal-associated pathogens in the environment is difficult due to their diversity and low concentration, therefore contamination is monitored using fecal indicator bacteria (FIB). FIB are present in the gastrointestinal tract of animals and provide an indication of the presence of fecal contamination. Escherichia coli (freshwater) and enterococci (fresh and salt water) are commonly used as FIB globally and in the United States. In sand, FIB are present at higher concentrations than in water, as sand protects FIB from environmental stressors such as UV or predation and provides easier access to nutrients. FIB that survive long-term and replicate in sand are termed “naturalized.” Naturalized populations can complicate the identification of recent fecal contamination, as FIB monitoring techniques cannot differentiate between naturalized FIB and those recently introduced through sources of fecal contamination. The ubiquity of FIB among animals and humans means it is also difficult to determine the source of contamination. Some sources of fecal contamination (human as compared to cow or bird fecal contamination) contain a higher number of pathogens that are likely to infect humans (adenovirus in humans or Cryptosporidium spp. from cows). Therefore, microbial source tracking (MST) was developed to include a suite of host-associated genes (markers) of fecal microorganisms that are specific to different animals. These markers can differentiate between sources of fecal contamination, supplementing FIB monitoring and providing a more accurate depiction of the fecal contamination picture. Few studies have investigated the presence of MST markers in sand or determined relationships between MST markers and FIB in sand. The human health risk associated with exposure to fecal contamination can be assessed by epidemiological studies; however, these studies are typically expensive and require specialized teams that may not be available to organizations with limited resources. Quantitative microbial risk assessment (QMRA) is a mathematical modeling framework used to estimate human health risk (the likelihood someone becomes ill) from exposure to pathogens under different environmental scenarios. QMRA consists of four steps: Problem formulation, exposure assessment, dose-response modeling, and risk characterization. Problem formulation involves establishing the framework (reference pathogen identification, exposure pathway, sources of contamination) that helps target the risk management needs to be addressed. The exposure assessment then determines the concentration and frequency of exposure by individuals to the reference pathogen(s) and exposure pathway identified during the problem formulation step, also known as a dose. The dose is then compared to a dose-response model (typically sourced from the literature) to determine the probability that the estimated dose would lead to illness in an individual. Risk characterization then quantifies the level of risk based on the modeled data. Reverse QMRA, used in this experiment, estimates the probabilities of pathogen concentrations that correspond with a risk threshold that is defined by stakeholders. Few QMRAs have been conducted in sand, therefore, this dissertation is focused on understanding the human health risk from exposure to pathogens at a tropical beach impacted by sewage. Jacó beach is a tropical beach located on the Pacific coast of Costa Rica and is a popular tourist destination. A water quality study determined the fecal contamination was severely impacting the water quality of the beach and placing individuals at risk of illness from exposure to contaminated waters. Understanding the human health risk from exposure to pathogens in beach sand, in addition to the human health risk from water, ultimately helps to improve beach management decisions and public health. In chapter one, the analytical sensitivity of quantitative polymerase chain reaction (qPCR) for Enterococcus in sand was compared for the slurry (suspension, agitation, membrane filtration of supernatant), versus two direct extraction methods using PowerSoil™ or PowerMax Soil™ kits at a freshwater and saltwater beach in Tampa, Fl, USA. We found the slurry method had the lowest limit of detection at 20–80 gene copies g-1 (wet weight), recovered significantly more DNA, and was the only method that detected Enterococcus by qPCR in all samples; therefore, the slurry method was exclusively used in subsequent experiments. The slurry method reflected the spatial variability of Enterococcus in individual transect samples. Mean recovery efficiency of the human-associated microbial source tracking marker HF183 from marine and freshwater beach sand spiked with wastewater was 100.8% and 64.1%, respectively, but varied between dilutions, indicating that the mixing protocol needs improvement. The objective of chapter two was to determine the extent of the influence a contaminated waterbody has on the concentration of microbes in beach sand. Upstream, downstream and ocean samples were collected on a transect in sand at 0 m (origin), 2.5 m and 5.0 m from the riverbank or swash zone. Samples were also collected in sediment. Samples were analyzed for the presence of the MST marker HF183 and FIB Enterococcus using the slurry method followed by DNA extraction and qPCR. Median concentrations of Enterococcus decreased as distance from the river or ocean increased. Enterococcus ranged from 1.46 x 104 to 8.11 x 103 gene copies 100 g-1 to the same distance. HF183 and Enterococcus were positively correlated at the riverbank/swash zone, but not at other subsites due to frequent failure to detect HF183. Sediment samples did not differ in HF183 or Enterococcus concentrations along the river and HF183 was not detected in the ocean sediment. The frequency of detection for HF183 was significantly greater among samples along the riverbank than samples collected at the 2.5 m and 5 m subsites. Polluted waterbodies can influence microbe concentrations in sand but the extent of the influence of a waterbody on MST markers in sand requires further study. Chapter three employed a reverse QMRA in Copey River at Jacó Beach to estimate the varying probability that corresponds with meeting a risk target. The risk target was defined as 36 cases of gastrointestinal illness per 1000 people (36/1000), the health target set in the 2012 recreational water quality guidelines for the United States Environmental Protection Agency (USEPA). Data were used to calculate whether concentrations of Salmonella, Adenovirus, and Giardia exceeded or were below concentrations needed to meet a risk of 36/1000. Samples at two sites, one upstream and one downstream of where beachgoers tended to recreate at the riverbank were collected six times over three weeks. Samples were analyzed for the presence of enterococci (FIB), the human-host associated MST marker HF183 and sewage-associated MST marker PMMoV, and pathogens Salmonella, adenovirus, and Giardia. Sand samples were collected at the riverbank and analyzed for the fecal indicator bacteria enterococci, the human-host associated microbial source tracking marker genes HF183, sewage-associated marker pepper mild mottle virus (PMMoV), and pathogens Salmonella, Giardia, and human adenovirus. Enterococci and Salmonella were cultured using the slurry method followed by USEPA standard methods. HF183, PMMoV, and adenovirus were detected by (RT)-qPCR. Giardia was detected using microscopy following USEPA standard methodology. Enterococci were detected in all samples and had a geometric mean of 3.25 x 103 GC 100 g-1. Geometric means of the MST markers HF183 and PMMoV were 1.19 x 103 and 1.64 x103 GC 100 g-1 and were detected in 83% and 33% of samples, respectively. Salmonella was detected in 66.6% of samples and had a geometric mean of 2.45 x 102 CFU 100 g-1. Giardia and adenovirus were detected only once each at 45 oocysts 100 g-1 and 1.91 x 102 GC 100 g-1. QMRA analysis showed that adenovirus and Giardia concentrations exceeded the levels that coincide with the risk threshold of 36/1000 while Salmonella fell between the 50th and 75th % percentile probability of meeting the risk threshold. Exposure to sewage-borne pathogens in sand could contribute to the risk of illness for recreational users of Jacó Beach and should be analyzed in tandem with pathogens in water to implement the best strategy for the protection of public health. To implement a comprehensive beach monitoring strategy, it is important to understand all the contextual factors that affect public health at a beach. Recreation in sand has been linked to gastrointestinal illness, however there are no regulatory guidelines or standard methods for analysis of fecal microorganisms in sand. This research has shown that a comprehensive beach monitoring strategy should include sand monitoring strategies in tandem with water monitoring strategies

Coliphage as an Indicator of the Quality of Beach Water to Protect the Health of Swimmers in Coastal Georgia

Background: Gastrointestinal disease affects millions of people in the United States and places a substantial economic burden upon healthcare systems. Recreational waters polluted with fecal material are a main source for transmission of gastrointestinal disease. Georgia beaches are monitored for the presence of fecal indicator bacteria, but these bacteria are not well associated with enteric viruses. The United States Environmental Protection Agency (US EPA) has recently proposed coliphage (a virus of Escherichia coli) as an alternative indicator of fecal contamination in recreational waters. The present study compares fecal indicator bacteria and coliphage concentrations at two Georgia beaches with adjacent creeks that have a history of pollution. Methods: For one year, samples and environmental data were collected from four sites on Jekyll Island, GA, during the peak swimming season and the off-season. Samples were processed using US EPA-approved methods for membrane filtration and plaque formation. Statistical analyses were performed using t-tests and Spearman correlations. Results: The highest numbers of enterococci and significant differences with coliphage were found at Saint Andrews Creek during the swimming season and the off-season. The enterococci concentrations at Clam Creek sites did not exceed recommended recreational water criteria. During the off-season, concentrations of enterococci and coliphages were different at Clam Creek sites, indicating a potential risk for presence of enteric virus when enterococci could not be detected. Conclusions: The US EPA has proposed to adapt coliphage concentrations as an alternative indicator of water pollution for routine beach monitoring nationally. The present study provides a background for adoption of this method in Georgia. Measures of enterococci do not provide sufficient information about the associated human health risk. Inclusion of these viral indicators will improve decision making for beach closures and for protection of the health of swimmers

Project PrEP: A Community Health Education Initiative

This presentation will describe the process of planning and implementing Project PrEP, a community health initiative to increase PrEP awareness among communities at high HIV risk in south Georgia, especially MSM. Intervention strategies, key findings, and lessons learned will be discussed

Using a combination of quantitative culture, molecular, and infrastructure data to rank potential sources of fecal contamination in Town Creek Estuary, North Carolina.

Estuarine water quality is declining worldwide due to increased tourism, coastal development, and a changing climate. Although well-established methods are in place to monitor water quality, municipalities struggle to use the data to prioritize infrastructure for monitoring and repair and to determine sources of contamination when they occur. The objective of this study was to assess water quality and prioritize sources of contamination within Town Creek Estuary (TCE), Beaufort, North Carolina, by combining culture, molecular, and geographic information systems (GIS) data into a novel contamination source ranking system. Water samples were collected from TCE at ten locations on eight sampling dates in Fall 2021 (n = 80). Microbiological water quality was assessed using US Environmental Protection Agency (U.S. EPA) approved culture-based methods for fecal indicator bacteria (FIB), including analysis of total coliforms (TC), Escherichia coli (EC), and Enterococcus spp. (ENT). The quantitative microbial source tracking (qMST) human-associated fecal marker, HF183, was quantified using droplet digital PCR (ddPCR). This information was combined with environmental data and GIS information detailing proximal sewer, septic, and stormwater infrastructure to determine potential sources of fecal contamination in the estuary. Results indicated FIB concentrations were significantly and positively correlated with precipitation and increased throughout the estuary following rainfall events (p < 0.01). Sampling sites with FIB concentrations above the U.S. EPA threshold also had the highest percentages of aged, less durable piping materials. Using a novel ranking system combining concentrations of FIB, HF183, and sewer infrastructure data at each site, we found that the two sites nearest the most aged sewage infrastructure and stormwater outflows were found to have the highest levels of measurable fecal contamination. This case study supports the inclusion of both traditional water quality measurements and local infrastructure data to support the current need for municipalities to identify, prioritize, and remediate failing infrastructure