Passive sampling of microbes in various water sources: A systematic review.

Traditional methods for monitoring pathogens in environmental waters have numerous drawbacks. Sampling approaches that are low-cost and time efficient that can capture temporal variation in microbial contamination are needed. Passive sampling of aquatic environments has shown promise as an alternative water monitoring technique for waterborne pathogens and microbial contaminants. The present systematic review aimed to compile and synthesize existing literature on the use of passive samplers for the monitoring of microbes in different water sources and identify research gaps. The review summarizes current knowledge on materials used for detection, deployment durations, analytical methods, quantification as well as benefits and limitations of passive sampling. This review found that electronegative nitrocellulose membrane filters are effective for both detection and quantification of viruses in wastewater, while gauze passive samplers have been effective for detecting bacterial targets in wastewater. There is a large knowledge gap in the use of passive samplers in a quantitative manner, especially for the back-calculation of water-column microbial concentrations or for correlation to outcomes of interest (e.g. prevalence rates). Further, there is very limited attention paid to the use of membrane filters for the monitoring of bacteria in any water source as well as a lack of studies utilizing passive sampling approaches for protozoa.

Burden of disease from contaminated drinking water in countries with high access to safely managed water: A systematic review.

The vast majority of residents of high-income countries (≥90%) reportedly have high access to safely managed drinking water. Owing perhaps to the widely held perception of near universal access to high-quality water services in these countries, the burden of waterborne disease in these contexts is understudied. This systematic review aimed to: identify population-scale estimates of waterborne disease in countries with high access to safely managed drinking water, compare methods to quantify disease burden, and identify gaps in available burden estimates. We conducted a systematic review of population-scale disease burden estimates attributed to drinking water in countries where ≥90% of the population has access to safely managed drinking water per official United Nations monitoring. We identified 24 studies reporting estimates for disease burden attributable to microbial contaminants. Across these studies, the median burden of gastrointestinal illness risks attributed to drinking water was ∼2,720 annual cases per 100,000 population. Beyond exposure to infectious agents, we identified 10 studies reporting disease burden-predominantly, cancer risks-associated with chemical contaminants. Across these studies, the median excess cancer cases attributable to drinking water was 1.2 annual cancer cases per 100,000 population. These median estimates slightly exceed WHO-recommended normative targets for disease burden attributable to drinking water and these results highlight that there remains important preventable disease burden in these contexts, particularly among marginalized populations. However, the available literature was scant and limited in geographic scope, disease outcomes, range of microbial and chemical contaminants, and inclusion of subpopulations (rural, low-income communities; Indigenous or Aboriginal peoples; and populations marginalized due to discrimination by race, ethnicity, or socioeconomic status) that could most benefit from water infrastructure investments. Studies quantifying drinking water-associated disease burden in countries with reportedly high access to safe drinking water, focusing on specific subpopulations lacking access to safe water supplies and promoting environmental justice, are needed.

A systematic review and meta-analysis of pathogen reduction in onsite sanitation systems.

The safe management of fecal sludge from the 3.4 billion people worldwide that use onsite sanitation systems can greatly reduce the global infectious disease burden. However, there is limited knowledge about the role of design, operational, and environmental factors on pathogen survival in pit latrines, urine diverting desiccation toilets, and other types of onsite toilets. We conducted a systematic literature review and meta-analysis to characterize pathogen reduction rates in fecal sludge, feces, and human excreta with respect to pH, temperature, moisture content, and the use of additives for desiccation, alkalinization, or disinfection. A meta-analysis of 1,382 data points extracted from 243 experiments described in 26 articles revealed significant differences between the decay rates and T99 values of pathogens and indicators from different microbial groups. The overall median T99 values were 4.8 days, 29 days, >341 days, and 429 days for bacteria, viruses, protozoan (oo)cysts, and Ascaris eggs, respectively. As expected, higher pH values, higher temperatures, and the application of lime all significantly predicted greater pathogen reduction rates but the use of lime by itself was more effective for bacteria and viruses than for Ascaris eggs, unless urea was also added. In multiple lab-scale experiments, the application of urea with enough lime or ash to reach a pH of 10 - 12 and a sustained concentration of 2,000 - 6,000 mg/L of non-protonated NH3-N reduced Ascaris eggs more rapidly than without urea. In general, the storage of fecal sludge for 6 months adequately controls hazards from viruses and bacteria, but much longer storage times or alkaline treatment with urea and low moisture or heat is needed to control hazards from protozoa and helminths. More research is needed to demonstrate the efficacy of lime, ash, and urea in the field. More studies of protozoan pathogens are also needed, as very few qualifying experiments were found for this group.

Study design and methods of the Wells and Enteric disease Transmission (WET) Trial: a randomised controlled trial.

INTRODUCTION: The burden of disease attributed to drinking water from private wells is not well characterised. The Wells and Enteric disease Transmission trial is the first randomised controlled trial to estimate the burden of disease that can be attributed to the consumption of untreated private well water. To estimate the attributable incidence of gastrointestinal illness (GI) associated with private well water, we will test if the household treatment of well water by ultraviolet light (active UV device) versus sham (inactive UV device) decreases the incidence of GI in children under 5 years of age. METHODS AND ANALYSIS: The trial will enrol (on a rolling basis) 908 families in Pennsylvania, USA, that rely on private wells and have a child 3 years old or younger. Participating families are randomised to either an active whole-house UV device or a sham device. During follow-up, families will respond to weekly text messages to report the presence of signs and symptoms of gastrointestinal or respiratory illness and will be directed to an illness questionnaire when signs/symptoms are present. These data will be used to compare the incidence of waterborne illness between the two study groups. A randomly selected subcohort submits untreated well water samples and biological specimens (stool and saliva) from the participating child in both the presence and absence of signs/symptoms. Samples are analysed for the presence of common waterborne pathogens (stool and water) or immunoconversion to these pathogens (saliva). ETHICS: Approval has been obtained from Temple University's Institutional Review Board (Protocol 25665). The results of the trial will be published in peer-reviewed journals. TRIAL REGISTRATION NUMBER: NCT04826991.

The effects of racism, social exclusion, and discrimination on achieving universal safe water and sanitation in high-income countries.

Drinking water and sanitation services in high-income countries typically bring widespread health and other benefits to their populations. Yet gaps in this essential public health infrastructure persist, driven by structural inequalities, racism, poverty, housing instability, migration, climate change, insufficient continued investment, and poor planning. Although the burden of disease attributable to these gaps is mostly uncharacterised in high-income settings, case studies from marginalised communities and data from targeted studies of microbial and chemical contaminants underscore the need for continued investment to realise the human rights to water and sanitation. Delivering on these rights requires: applying a systems approach to the problems; accessible, disaggregated data; new approaches to service provision that centre communities and groups without consistent access; and actionable policies that recognise safe water and sanitation provision as an obligation of government, regardless of factors such as race, ethnicity, gender, ability to pay, citizenship status, disability, land tenure, or property rights.

Passive sampling, a practical method for wastewater-based surveillance of SARS-CoV-2.

In search of practical and affordable tools for wastewater-based surveillance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), three independent field experiments were conducted using three passive sampler sorbents (electronegative membrane, cotton bud, and gauze) in Guelph, Ontario, Canada. Total daily cases during this study ranged from 2 to 17/100,000 people and 43/54 traditionally collected wastewater samples were positive for SARS-CoV-2 with mean detectable concentrations ranging from 8.4 to 1780 copies/ml. Viral levels on the passive samplers were assessed after 4, 8, 24, 48, 72, and 96 hrs of deployment in the wastewater and 43/54 membrane, 42/54 gauze, and 27/54 cotton bud samples were positive. A linear accumulation rate of SARS-CoV-2 on the membranes was observed up to 48 hours, suggesting the passive sampler could adequately reflect wastewater levels for up to two days of deployment. Due the variability in accumulation observed for the cotton buds and gauzes, and the pre-processing steps required for the gauzes, we recommend membrane filters as a simple cost-effective option for wastewater-based surveillance of SARS-CoV-2.

Microbial water quality improvement associated with transitioning from intermittent to continuous water supply in Nagpur, India.

Nearly half a billion people living in Indian cities receive their drinking water from an intermittent water supply (IWS), which can be associated with degraded water quality and risk of waterborne disease. The municipal water supply in Nagpur, India is transitioning from intermittent to continuous supply in phases. We conducted cross-sectional sampling to compare microbial water quality under IWS and continuous water supply (CWS) in Nagpur. In 2015 and 2017, we collected 146 grab samples and 90 large-volume dead-end ultrafiltration (DEUF) samples (total volume: 6,925 liters). In addition to measuring traditional water quality parameters, we also assayed DEUF samples by droplet digital PCR (ddPCR) for waterborne pathogen gene targets. At household taps served by IWS, we detected targets from enterotoxigenic E. coli, Shigella spp./enteroinvasive E. coli, norovirus GI and GII, adenovirus A-F, Cryptosporidium spp., and Giardia duodenalis. We observed a significant increase in the proportion of grab samples positive for culturable E. coli (p = 0.0007) and DEUF concentrates positive for waterborne pathogen gene targets (p = 0.0098) at household taps served by IWS compared to those served by CWS. IWS continues to be associated with fecal contamination, and, in this study, with increased prevalence of molecular evidence of waterborne pathogens. These findings add mounting evidence that, despite the presence of piped on premise infrastructure, IWS is less likely to meet the requirements for safely-managed drinking water as defined by the Sustainable Development Goals. Importantly, these findings demonstrate the transition from IWS to CWS in Nagpur is yielding meaningful improvements in microbial water quality.

The Impact of Pipe Material on the Diversity of Microbial Communities in Drinking Water Distribution Systems.

As many cities around the world face the prospect of replacing aging drinking water distribution systems (DWDS), water utilities must make careful decisions on new pipe material (e.g., cement-lined or PVC) for these systems. These decisions are informed by cost, physical integrity, and impact on microbiological and physicochemical water quality. Indeed, pipe material can impact the development of biofilm in DWDS that can harbor pathogens and impact drinking water quality. Annular reactors (ARs) with cast iron and cement coupons fed with chloraminated water from a municipal DWDS were used to investigate the impact of pipe material on biofilm development and composition over 16 months. The ARs were plumbed as closely as possible to the water main in the basement of an academic building to simulate distribution system conditions. Biofilm communities on coupons were characterized using 16S rRNA sequencing. In the cast iron reactors, ß-proteobacteria, Actinobacteria, and α-proteobacteria were similarly relatively abundant (24.1, 22.5, and 22.4%, respectively) while in the cement reactors, α-proteobacteria and Actinobacteria were more relatively abundant (36.3 and 35.2%, respectively) compared to ß-proteobacteria (12.8%). Mean alpha diversity (estimated with Shannon H and Faith's Phylogenetic Difference indices) was greater in cast iron reactors (Shannon: 5.00 ± 0.41; Faith's PD: 15.40 ± 2.88) than in cement reactors (Shannon: 4.16 ± 0.78; Faith's PD: 13.00 ± 2.01). PCoA of Bray-Curtis dissimilarities indicated that communities in cast iron ARs, cement ARs, bulk distribution system water, and distribution system pipe biofilm were distinct. The mean relative abundance of Mycobacterium spp. was greater in the cement reactors (34.8 ± 18.6%) than in the cast iron reactors (21.7 ± 11.9%). In contrast, the mean relative abundance of Legionella spp. trended higher in biofilm from cast iron reactors (0.5 ± 0.7%) than biofilm in cement reactors (0.01 ± 0.01%). These results suggest that pipe material is associated with differences in the diversity, bacterial composition, and opportunistic pathogen prevalence in biofilm of DWDS.

Why pathogens matter for meeting the united nations' sustainable development goal 6 on safely managed water and sanitation.

Water and wastewater utilities, water and sanitation hygiene (WASH) practitioners, and regulating bodies, particularly in developing nations, rely heavily on indicator microorganisms, as opposed to pathogens, for much of their regulatory decisions. This commentary illustrates the importance of considering pathogens and not relying only on indicator organisms when making decisions regarding water and sanitation, especially with respect to meeting the current targets of the Sustainable Development Goal (SDG) 6. We use quantitative microbial risk assessment (QMRA) to present three common scenarios that WASH and public health practitioners encounter to illustrate our point. These include 1) chlorination of surface water for drinking, 2) land application of latrine waste as a fertilizer, and 3) recreation/domestic use of surface waters impacted by wastewater discharge. We show that the calculated probabilities of risk of infection are statistically significantly higher when using treatment/survival information for pathogens versus using indicator species data. Thus, demonstrating that relying solely on indicators for sanitation decision making is inadequate if we truly want to achieve the SDG6 targets of safely managed water and sanitation services.

Septic Systems and Rainfall Influence Human Fecal Marker and Indicator Organism Occurrence in Private Wells in Southeastern Pennsylvania.

In the United States, approximately 48 million people are served by private wells. Unlike public water systems, private well water quality is not monitored, and there are few studies on the extent and sources of contamination of private wells. We extensively investigated five private wells to understand the variability in microbial contamination, the role of septic systems as sources of contamination, and the effect of rainfall on well water quality. From 2016 to 2017, weekly or biweekly samples (n = 105) were collected from five private wells in rural Pennsylvania. Samples were tested for general water quality parameters, conventional and sewage-associated microbial indicators, and human pathogens. Total coliforms, human Bacteroides (HF183), and pepper mild mottle virus were detected at least once in all wells. Regression revealed significant relationships between HF183 and rainfall 8-14 days prior to sampling and between total coliforms and rainfall 8-14 or 0-14 days prior to sampling. Dye tracer studies at three wells confirmed the impact of household septic systems on well contamination. Microbiological measurements, chemical water quality data, and dye tracer tests provide evidence of human fecal contamination in the private wells studied, suggesting that household septic systems are the source of this contamination.

Private Wells and Rural Health: Groundwater Contaminants of Emerging Concern.

PURPOSE OF REVIEW: Approximately 12% of the population in the US and Canada rely on federally unregulated private wells, which are common in rural areas and may be susceptible to microbiological and chemical contamination. This review identifies and summarizes recent findings on contaminants of emerging concern in well water across the US and Canada. RECENT FINDINGS: Private well water quality modeling is complicated by the substantial variability in contamination sources, well construction, well depth, and the hydrogeology of the environment surrounding the well. Temporal variation in contaminant levels in wells suggests the need for monitoring efforts with greater spatial and temporal coverage. More extensive private well monitoring will help identify wells at greater risk of contamination, and in turn, public health efforts can focus on education and outreach to improve monitoring, maintaining, and treating private wells in these communities. Community interventions need to be coupled with stricter regulations and financing mechanisms that can support and protect private well owners.

School-based, blacklight handwashing program can improve handwashing quality and knowledge among pre-school aged children.

In the United States, lack of proper handwashing is associated with respiratory and gastrointestinal illnesses. Interventions to improve handwashing practices have led to an increase in handwashing knowledge and behavior, and decreases in gastrointestinal illnesses. Most studies have evaluated their interventions in the context of reported handwashing rates by observation, reduction of illnesses, as well as reduced absences, however none of these studies examined handwashing quality or knowledge as outcome measures. The objective of this paper is to present the results from a handwashing program with a special focus on the evaluation methods. A pre-post design was used to evaluate a handwashing program that took place in two pre-schools the northeast of the United States. The program utilized a black light technology to demonstrate to children the importance of good quality. The evaluation consisted of assessing knowledge and quality of handwashing using a linear puzzle and individual handwashing observation, respectively. Students from both schools improved on both knowledge and quality over time (p-values 0.071 and <0.001, respectively). The present study demonstrates that the use of black light technology as an educational tool may help to improve handwashing quality among pre-school aged children, even after only one instructional session.

Translating pathogen knowledge to practice for sanitation decision-making.

Sanitation planners make complex decisions in the delivery of sanitation services to achieve health outcomes. We present findings from a stakeholder engagement workshop held in Kampala, Uganda, to educate, interact with, and solicit feedback from participants on how the relevant scientific literature on pathogens can be made more accessible to practitioners to support decision-making. We targeted Water, Sanitation and Hygiene (WASH) practitioners involved in different levels of service delivery. Practitioners revealed that different sanitation planning tools are used to inform decision-making; however, most of these tools are not user-friendly or adapted to meet their needs. Most stakeholders (68%) expressed familiarity with pathogens, yet less than half (46%) understood that fecal coliforms were bacteria and used as indicators for fecal pollution. A number of stakeholders were unaware that fecal indicator bacteria do not behave and persist the same as helminths, protozoa, or viruses, making fecal indicator bacteria inadequate for assessing pathogen reductions for all pathogen groups. This suggests a need for awareness and capacity development around pathogens found in excreta. The findings underscore the importance to engage stakeholders in the development of support tools for sanitation planning and highlighted broader opportunities to bridge science with practice in the WASH sector.

Using decision trees to understand the influence of individual- and neighborhood-level factors on urban diabetes and asthma.

OBJECTIVE: To determine the influence of individual and neighborhood factors that combined are associated with asthma and diabetes in a sample of urban Philadelphians using data mining, a novel technique in public health research. METHODS: We obtained secondary data collected between May 2011 and November 2014 on individual's health and perception of neighborhood characteristics (N = 450) and Philadelphia LandCare Program data that provided relevant environmental data for the analysis (N = 676). RapidMiner open access data mining software was used to perform decision tree analyses. RESULTS: Individual- and neighborhood-level environmental factors were intricately related in the decision tree models, having varying influence on the outcomes of asthma and diabetes. The decision trees had high specificity (95-100) and classified factors that were associated with an absence of disease (diabetes/asthma). CONCLUSION: Improved neighborhood-level conditions related to social and physical disorder were consistently found to be associated with an absence of both asthma and diabetes in this urban population. POLICY IMPLICATIONS: This study illustrates the potential utility of applying data mining techniques for understanding complex public health issues.

Bacterial Contamination on Latrine Surfaces in Community and Household Latrines in Kathmandu, Nepal.

A lack of sanitation infrastructure is a major contributor to the global burden of diarrheal disease, particularly in low-income countries. Access to basic sanitation was identified as part of the 2015 United Nations Sustainable Development Goals. However, current definitions of "basic" sanitation infrastructure exclude community or shared sanitation, due to concerns around safety, equity, and cleanliness. The purpose of this study was to measure and compare bacterial contamination on community and household latrine surfaces in Kathmandu, Nepal. One hundred and nineteen swab samples were collected from two community and five household latrines sites. Community latrine samples were taken before and after daily cleaning, while household samples were collected at midday, to reflect normal conditions. Concentrations of total coliforms and Escherichia coli were measured using membrane filtration methods. Results found almost no differences between bacterial contamination on latrine surfaces in community and household latrines, with the exception of latrine slabs/seats that were more contaminated in the community latrines under dirty conditions. The study also identified surfaces with higher levels of contamination. Findings demonstrated that well-maintained community latrines may be as clean, or cleaner, than household latrines and support the use of community latrines for improving access to sanitation infrastructure in a low-income country setting.

Human Bacteroides and total coliforms as indicators of recent combined sewer overflows and rain events in urban creeks.

Combined sewer overflows (CSOs) are a known source of human fecal pollution and human pathogens in urban water bodies, which may present a significant public health threat. To monitor human fecal contamination in water, bacterial fecal indicator organisms (FIOs) are traditionally used. However, because FIOs are not specific to human sources and do not correlate with human pathogens, alternative fecal indicators detected using qPCR are becoming of interest to policymakers. For this reason, this study measured correlations between the number and duration of CSOs and mm of rainfall, concentrations of traditional FIOs and alternative indicators, and the presence of human pathogens in two urban creeks. Samples were collected May-July 2016 and analyzed for concentrations of FIOs (total coliforms and E. coli) using membrane filtration as well as for three alternative fecal indicators (human Bacteroides HF183 marker, human polyomavirus (HPoV), pepper mild mottle virus (PMMoV)) and nine human pathogens using qPCR. Four of the nine pathogens analyzed were detected at these sites including adenovirus, Enterohemorrhagic E. coli, norovirus, and Salmonella. Among all indicators studied, human Bacteroides and total coliforms were significantly correlated with recent CSO and rainfall events, while E. coli, PMMoV, and HPoV did not show consistent significant correlations. Further, human Bacteroides were a more specific indicator, while total coliforms were a more sensitive indicator of CSO and rainfall events. Results may have implications for the use and interpretation of these indicators in future policy or monitoring programs.

Current Stormwater Harvesting Guidelines Are Inadequate for Mitigating Risk from Campylobacter During Nonpotable Reuse Activities.

Campylobacter is a pathogen frequently detected in urban stormwater worldwide. It is one of the leading causes of enteric disease in many developed countries and is the leading cause of enteric disease in Australia. Prior to harvesting stormwater, adequate treatment is necessary to mitigate risks derived from such harmful pathogens. The goal of this research was to estimate the health risks associated with the exposure to Campylobacter when harvesting urban stormwater for toilet flushing and irrigation activities, and the role treatment options play in limiting risks. Campylobacter data collected from several urban stormwater systems in Victoria, Australia, were the inputs of a Quantitative Microbial Risk Assessment model. The model included seven treatment scenarios, spanning wetlands, biofilters, and more traditional treatment trains including those recommended by the Australian Guidelines for Water Recycling. According to our modeling and acceptable risk thresholds, only two treatment scenarios could supply water of sufficient quality for toilet flushing and irrigation end-uses: (1) using stormwater biofilters coupled with UV-treatment and (2) a more conventional coagulation, filtration, UV, and chlorination treatment plant. Importantly, our modeling results suggest that current guidelines in place for stormwater reuse are not adequate for protecting against exposure to Campylobacter. However, more research is required to better define whether the Campylobacter detectable in stormwater are pathogenic to humans.

A Systematic Review: Costing and Financing of Water, Sanitation, and Hygiene (WASH) in Schools.

Despite the success of recent efforts to increase access to improved water, sanitation, and hygiene (WASH) globally, approximately one-third of schools around the world still lack adequate WASH services. A lack of WASH in schools can lead to the spread of preventable disease and increase school absences, especially among women. Inadequate financing and budgeting has been named as a key barrier for integrating successful and sustainable WASH programs into school settings. For this reason, the purpose of this review is to describe the current knowledge around the costs of WASH components as well as financing models that could be applied to WASH in schools. Results show a lack of information around WASH costing, particularly around software elements as well as a lack of data overall for WASH in school settings as compared to community WASH. This review also identifies several key considerations when designing WASH budgets or selecting financing mechanisms. Findings may be used to advise future WASH in school programs.

Elevated levels of iron in groundwater in Prey Veng province in Cambodia: a possible factor contributing to high iron stores in women.

Iron is a natural element found in food, water and soil and is essential for human health. Our aim was to determine the levels of iron and 25 other metals and trace elements in groundwater from 22 households in Prey Veng, Cambodia. Water analyses were conducted using inductively coupled plasma-mass spectrometry and optical emission spectrometry. Compared to the 2011 World Health Organization guidelines for drinking water quality, aluminum, iron and manganese exceeded maximum levels (in 4.5, 72.7 and 40.9% of samples, respectively). Compared to the 2004 Cambodian drinking water quality standards, iron and manganese exceeded maximum levels (in 59.1 and 36.4% of samples, respectively). We found no evidence of arsenic contamination. Guidelines for iron were established primarily for esthetic reasons (e.g. taste), whereas other metals and elements have adverse effects associated with toxicity. Iron in groundwater ranged from 134 to 5,200 µg/L (mean ∼1,422 µg/L). Based on a daily consumption of 3 L groundwater, this equates to ∼0.4-15.6 mg iron (mean ∼4.3 mg/day), which may be contributing to high iron stores and the low prevalence of iron deficiency anemia in Prey Veng women. Elevated levels of manganese in groundwater are a concern and warrant further investigation.

Groundwater source contamination mechanisms: physicochemical profile clustering, risk factor analysis and multivariate modelling.

An integrated domestic well sampling and "susceptibility assessment" programme was undertaken in the Republic of Ireland from April 2008 to November 2010. Overall, 211 domestic wells were sampled, assessed and collated with local climate data. Based upon groundwater physicochemical profile, three clusters have been identified and characterised by source type (borehole or hand-dug well) and local geological setting. Statistical analysis indicates that cluster membership is significantly associated with the prevalence of bacteria (p=0.001), with mean Escherichia coli presence within clusters ranging from 15.4% (Cluster-1) to 47.6% (Cluster-3). Bivariate risk factor analysis shows that on-site septic tank presence was the only risk factor significantly associated (p<0.05) with bacterial presence within all clusters. Point agriculture adjacency was significantly associated with both borehole-related clusters. Well design criteria were associated with hand-dug wells and boreholes in areas characterised by high permeability subsoils, while local geological setting was significant for hand-dug wells and boreholes in areas dominated by low/moderate permeability subsoils. Multivariate susceptibility models were developed for all clusters, with predictive accuracies of 84% (Cluster-1) to 91% (Cluster-2) achieved. Septic tank setback was a common variable within all multivariate models, while agricultural sources were also significant, albeit to a lesser degree. Furthermore, well liner clearance was a significant factor in all models, indicating that direct surface ingress is a significant well contamination mechanism. Identification and elucidation of cluster-specific contamination mechanisms may be used to develop improved overall risk management and wellhead protection strategies, while also informing future remediation and maintenance efforts.