Drinking water sources, quality, and associated health outcomes in Appalachian Virginia: A risk characterization study in two counties.

OBJECTIVES: In the US, violations of drinking water regulations are highest in lower-income rural areas overall, and particularly in Central Appalachia. However, data on drinking water use, quality, and associated health outcomes in rural Appalachia are limited. We sought to assess public and private drinking water sources and associated risk factors for waterborne pathogen exposures for individuals living in rural regions of Appalachian Virginia. METHODS: We administered surveys and collected tap water, bottled water, and saliva samples in lower-income households in two adjacent rural counties in southwest Virginia (bordering Kentucky and Tennessee). Water samples were tested for pH, temperature, conductivity, total coliforms, E. coli, free chlorine, nitrate, fluoride, heavy metals, and specific pathogen targets. Saliva samples were analyzed for antibody responses to potentially waterborne infections. We also shared water analysis results with households. RESULTS: We enrolled 33 households (83 individuals), 82% (n = 27) with utility-supplied water and 18% with private wells (n = 3) or springs (n = 3). 58% (n = 19) reported household incomes of <$20,000/year. Total coliforms were detected in water samples from 33% (n = 11) of homes, E. coli in 12%, all with wells or springs (n = 4), and Aeromonas, Campylobacter, and Enterobacter in 9%, all spring water (n = 3). Diarrhea was reported for 10% of individuals (n = 8), but was not associated with E. coli detection. 34% (n = 15) of saliva samples had detectable antibody responses for Cryptosporidium spp., C. jejuni, and Hepatitis E. After controlling for covariates and clustering, individuals in households with septic systems and straight pipes had significantly higher likelihoods of antibody detection (risk ratios = 3.28, 95%CI = 1.01-10.65). CONCLUSIONS: To our knowledge, this is the first study to collect and analyze drinking water samples, saliva samples, and reported health outcome data from low-income households in Central Appalachia. Our findings indicate that utility-supplied water in this region was generally safe, and individuals in low-income households without utility-supplied water or sewerage have higher exposures to waterborne pathogens.

Incidence of per- and polyfluoroalkyl substances (PFAS) in private drinking water supplies in Southwest Virginia, USA.

Per- and polyfluoroalkyl substances (PFAS) are a class of man-made contaminants of human health concern due to their resistance to degradation, widespread environmental occurrence, bioaccumulation in living organisms, and potential negative health impacts. Private drinking water supplies may be uniquely vulnerable to PFAS contamination in impacted areas, as these systems are not protected under federal regulations and often include limited treatment or remediation, if contaminated, prior to use. The goal of this study was to determine the incidence of PFAS contamination in private drinking water supplies in two counties in Southwest Virginia, USA (Floyd and Roanoke) that share similar bedrock geologies, are representative of different state Department of Health risk categories, and to examine the potential for reliance on citizen-science based strategies for sample collection in subsequent efforts. Samples for inorganic ions, bacteria, and PFAS analysis were collected on separate occasions by participants and experts at the home drinking water point of use (POU) for comparison. Experts also collected outside tap samples for analysis of 30 PFAS compounds. At least one PFAS was detectable in 95 % of POU samples collected (n = 60), with a mean total PFAS concentration of 23.5 ± 30.8 ppt. PFOA and PFOS, two PFAS compounds which presently have EPA health advisories, were detectable in 13 % and 22 % of POU samples, respectively. On average, each POU sample contained >3 PFAS compounds, and one sample contained as many as 8 compounds, indicating that exposure to a mixture of PFAS in drinking water may be occurring. Although there were significant differences in total PFAS concentrations between expert and participant collected samples (Wilcoxon, alpha = 0.05), collector bias was inconsistent, and may be due to the time of day of sampling (i.e. morning, afternoon) or specific attributes of a given home. Further research is required to resolve sources of intra-sample variability.

Making waves: The benefits and challenges of responsibly implementing wastewater-based surveillance for rural communities.

The sampling and analysis of sewage for pathogens and other biomarkers offers a powerful tool for monitoring and understanding community health trends and potentially predicting disease outbreaks. Since the early months of the COVID-19 pandemic, the use of wastewater-based testing for public health surveillance has increased markedly. However, these efforts have focused on urban and peri­urban areas. In most rural regions of the world, healthcare service access is more limited than in urban areas, and rural public health agencies typically have less disease outcome surveillance data than their urban counterparts. The potential public health benefits of wastewater-based surveillance for rural communities are therefore substantial - though so too are the methodological and ethical challenges. For many rural communities, population dynamics and insufficient, aging, and inadequately maintained wastewater collection and treatment infrastructure present obstacles to the reliable and responsible implementation of wastewater-based surveillance. Practitioner observations and research findings indicate that for many rural systems, typical implementation approaches for wastewater-based surveillance will not yield sufficiently reliable or actionable results. We discuss key challenges and potential strategies to address them. However, to support and expand the implementation of responsible, reliable, and ethical wastewater-based surveillance for rural communities, best practice guidelines and standards are needed.

Microbiological and chemical drinking water contaminants and associated health outcomes in rural Appalachia, USA: A systematic review and meta-analysis.

In rural areas of the United States, an estimated ~1.8 million people lack reliable access to safe drinking water. Considering the relative dearth of information on water contamination and health outcomes in Appalachia, we conducted a systematic review of studies of microbiological and chemical drinking water contamination and associated health outcomes in rural Appalachia. We pre-registered our protocols, limiting eligibility to primary data studies published from 2000 to 2019, and searched four databases (PubMed, EMBASE, Web of Science, and the Cochrane Library). We used qualitative syntheses, meta-analyses, risk of bias analysis, and meta-regression to assess reported findings, with reference to US EPA drinking water standards. Of the 3452 records identified for screening, 85 met our eligibility criteria. 93 % of eligible studies (n = 79) used cross-sectional designs. Most studies were conducted in Northern (32 %, n = 27) and North Central (24 %, n = 20) Appalachia, and only 6 % (n = 5) were conducted exclusively in Central Appalachia. Across studies, E. coli were detected in 10.6 % of samples (sample-size-weighted mean percentage from 4671 samples, 14 publications). Among chemical contaminants, sample-size-weighted mean concentrations for arsenic were 0.010 mg/L (n = 21,262 samples, 6 publications), and 0.009 mg/L for lead (n = 23,259, 5 publications). 32 % (n = 27) of studies assessed health outcomes, but only 4.7 % (n = 4) used case-control or cohort designs (all others were cross-sectional). The most commonly reported outcomes were detection of PFAS in blood serum (n = 13), gastrointestinal illness (n = 5), and cardiovascular-related outcomes (n = 4). Of the 27 studies that assessed health outcomes, 62.9 % (n = 17) appeared to be associated with water contamination events that had received national media attention. Overall, based on the number and quality of eligible studies identified, we could not reach clear conclusions about the state of water quality, or its impacts on health, in any of Appalachia's subregions. More epidemiologic research is needed to understand contaminated water sources, exposures, and potentially associated health outcomes in Appalachia.

Land cover and community water system characteristics as predictors of Safe Drinking Water Act violations in Central Appalachia, USA.

Upstream anthropogenic land cover can degrade source drinking water quality and thereby inhibit the ability of a community water system to provide safe drinking water. This study aimed to predict differences in Safe Drinking Water Act (SDWA) compliance between water systems based on upstream land cover in Central Appalachia and to examine whether national trends correlating violations with system size and source type were relevant for this region. Multiple generalized linear mixed models assessed relationships between SDWA violations and the distance weighted land cover proportions associated with the water system's contributing source watershed, as well as county economic status, system size, and water source. Results indicate that rates of monitoring and reporting violations were significantly higher for smaller water systems in more economically distressed counties. Interestingly, increases in surface mining landuse and high density development decreased monitoring and reporting violations, which may reflect impacts of associated economic development. Increases in low intensity development increased the likelihood of health-based violations. To protect public health, community managers should consider source water protection and/or upgrading drinking water system treatment capacity prior to developing previously undeveloped areas and further motivate compliance with monitoring and reporting requirements.

Examining the association between safe drinking water act violations and adverse birth outcomes in Virginia.

In 1974, the United States established the Safe Drinking Water Act (SDWA) to protect consumers from potential exposure to drinking water contaminants associated with health risks. Each contaminant is assigned a health-based standard meant to reflect the maximum level at which an adverse human health outcome is unlikely; measurements beyond that level have greater potential to result in adverse health outcomes. Although there is extensive research on human health implications following water contaminant exposure, few studies have specifically examined associations between fetal health and municipal drinking water violations. Therefore, the objective of this study is to assess whether SDWA drinking water violations are associated with fetal health outcomes, including preterm birth (PTB), low birth weight (LBW), and term-low birth weight (tLBW), in the Commonwealth of Virginia. Singleton births (n = 665,984) occurring between 2007 and 2015 in Virginia were geocoded and assigned to a corresponding estimated water service area. Health-based (HB) and monitoring and reporting (MR) violations for 12 contaminants were acquired from the US EPA Safe Drinking Water Information System, with exposure defined at the approximate service area level to limit exposure misclassification. A logistic regression model for each birth outcome assessed potential relationships with SDWA violations. When examining the association between individual MR violations and birth outcomes, Nitrate-Nitrite (OR = 1.10; 95% CI = 1.02, 1.18, P = 0.01) was positively associated with PTB and the total coliform rule was negatively associated with tLBW (OR = 0.93; 95% CI = 0.87, 1.00, P = 0.04). These findings indicate that a lack of regular monitoring and reporting by water providers (resulting in monitoring and reporting violations) may be concealing health-based violations as these health concerns cannot be revealed without testing, suggesting a need for additional technical, managerial, and financial support to enable often-underfunded water systems to adhere to monitoring and reporting requirements meant to protect public health.

Bottled and Well Water Quality in a Small Central Appalachian Community: Household-Level Analysis of Enteric Pathogens, Inorganic Chemicals, and Health Outcomes in Rural Southwest Virginia.

Consumption of unsafe drinking water is associated with a substantial burden of disease globally. In the US, ~1.8 million people in rural areas lack reliable access to safe drinking water. Our objective was to characterize and assess household-level water sources, water quality, and associated health outcomes in Central Appalachia. We collected survey data and water samples (tap, source, and bottled water) from consenting households in a small rural community without utility-supplied water in southwest Virginia. Water samples were analyzed for physicochemical parameters, total coliforms, E. coli, nitrate, sulfate, metals (e.g., arsenic, cadmium, lead), and 30+ enteric pathogens. Among the 69% (n = 9) of households that participated, all had piped well water, though 67% (n = 6) used bottled water as their primary drinking water source. Total coliforms were detected in water samples from 44.4% (n = 4) of homes, E. coli in one home, and enteric pathogens (Aeromonas, Campylobacter, Enterobacter) in 33% (n = 3) of homes. Tap water samples from 11% (n = 1) of homes exceeded the EPA MCL for nitrate, and 33% (n = 3) exceeded the EPA SMCL for iron. Among the 19 individuals residing in study households, reported diarrhea was 25% more likely in homes with measured E. coli and/or specific pathogens (risk ratio = 1.25, cluster-robust standard error = 1.64, p = 0.865). Although our sample size was small, our findings suggest that a considerable number of lower-income residents without utility-supplied water in rural areas of southwest Virginia may be exposed to microbiological and/or chemical contaminants in their water, and many, if not most, rely on bottled water as their primary source of drinking water.

Identifying exposure pathways mediating adverse birth outcomes near active surface mines in Central Appalachia.

Background: Previous work has determined an association between proximity to active surface mining within Central Appalachia and an increased risk of preterm birth (PTB) and low birthweight (LBW). Multiple potential exposure pathways may exist; however, including inhalation of particulate matter (airshed exposure), or exposure to impacted surface waters (watershed exposure). We hypothesize that this relationship is mediated by exposure to contaminants along one or both of these pathways. Methods: We geolocated 194,084 birth records through health departments in WV, KY, VA, and TN between 1990 and 2015. We performed a mediation analysis, iteratively including within our models: (a) the percent of active surface mining within 5 km of maternal residence during gestation; (b) the cumulative surface mining airshed trajectories experienced during gestation; and (c) the percent of active surface mining occurring within the watershed of residency during gestation. Results: Our baseline models found that active surface mining was associated with an increased odds of PTB (1.09, 1.05-1.13) and LBW (1.06, 1.02-1.11), controlling for individual-level predictors. When mediators were added to the baseline model, the association between active mining and birth outcomes became nonsignificant (PTB: 0.48, 0.14-1.58; LBW 0.78, 0.19-3.00), whereas the association between PTB and LBW remained significant by airshed exposure (PTB: 1.14, 1.11-1.18; LBW: 1.06, 1.03-1.10). Conclusions: Our results found that surface mining airsheds at least partially explained the association between active mining and adverse birth outcomes, consistent with a hypothesis of mediation, while mediation via the watershed pathway was less evident.

Approximating Community Water System Service Areas to Explore the Demographics of SDWA Compliance in Virginia.

Although the United States Safe Drinking Water Act (SDWA) theoretically ensures drinking water quality, recent studies have questioned the reliability and equity associated with community water system (CWS) service. This study aimed to identify SDWA violation differences (i.e., monitoring and reporting (MR) and health-based (HB)) between Virginia CWSs given associated service demographics, rurality, and system characteristics. A novel geospatial methodology delineated CWS service areas at the zip code scale to connect 2000 US Census demographics with 2006-2016 SDWA violations, with significant associations determined via negative binomial regression. The proportion of Black Americans within a service area was positively associated with the likelihood of HB violations. This effort supports the need for further investigation of racial and socioeconomic disparities in access to safe drinking water within the United States in particular and offers a geospatial strategy to explore demographics in other settings where data on infrastructure extents are limited. Further interdisciplinary efforts at multiple scales are necessary to identify the entwined causes for differential risks in adverse drinking water quality exposures and would be substantially strengthened by the mapping of official CWS service boundaries.

Integrated Metagenomic Assessment of Multiple Pre-harvest Control Points on Lettuce Resistomes at Field-Scale.

An integrated understanding of factors influencing the occurrence, distribution, and fate of antibiotic resistance genes (ARGs) in vegetable production systems is needed to inform the design and development of strategies for mitigating the potential for antibiotic resistance propagation in the food chain. The goal of the present study was to holistically track antibiotic resistance and associated microbiomes at three distinct pre-harvest control points in an agroecosystem in order to identify the potential impacts of key agricultural management strategies. Samples were collected over the course of a single growing season (67 days) from field-scale plots amended with various organic and inorganic amendments at agronomic rates. Dairy-derived manure and compost amendment samples (n = 14), soil samples (n = 27), and lettuce samples (n = 12) were analyzed via shotgun metagenomics to assess multiple pre-harvest factors as hypothetical control points that shape lettuce resistomes. Pre-harvest factors of interest included manure collection during/post antibiotic use, manure composting, and soil amended with organic (stockpiled manure/compost) versus chemical fertilizer. Microbial community resistome and taxonomic compositions were unique from amendment to soil to lettuce surface according to dissimilarity analysis. The highest resistome alpha diversity (i.e., unique ARGs, n = 642) was detected in amendment samples prior to soil application, while the composted manure had the lowest total ARG relative abundance (i.e., 16S rRNA gene-normalized). Regardless of amendment type, soils acted as an apparent ecological buffer, i.e., soil resistome and taxonomic profiles returned to background conditions 67 d-post amendment application. Effects of amendment conditions surprisingly re-emerged in lettuce phyllosphere resistomes, with the highest total ARG relative abundances recovered on the surface of lettuce plants grown in organically-fertilized soils (i.e., compost- and manure-amended soils). Co-occurrence analysis identified 55 unique ARGs found both in the soil amendments and on lettuce surfaces. Among these, arnA and pmrF were the most abundant ARGs co-occurring with mobile genetic elements (MGE). Other prominent ARG-MGE co-occurrences throughout this pre-harvest lettuce production chain included: TetM to transposon (Clostridiodies difficile) in the manure amendment and TriC to plasmid (Ralstonia solanacearum) on the lettuce surfaces. This suggests that, even with imposing manure management and post-amendment wait periods in agricultural systems, ARGs originating from manure can still be found on crop surfaces. This study demonstrates a comprehensive approach to identifying key control points for the propagation of ARGs in vegetable production systems, identifying potential ARG-MGE combinations that could inform future surveillance. The findings suggest that additional pre-harvest and potentially post-harvest interventions may be warranted to minimize risk of propagating antibiotic resistance in the food chain.

Freshwater salinization increases survival of Escherichia coli and risk of bacterial impairment.

Elevated levels of Escherichia coli (E. coli) are responsible for more designated freshwater stream impairments than any other contaminant in the United States. E. coli are intentionally used as a sentinel of fecal contamination for freshwaters because previous research indicates that salt concentrations in brackish or marine waters reduce E. coli survival, rendering it a less effective indicator of public health risks. Given increasing evidence of freshwater salinization associated with upland anthropogenic land-use, understanding the effects on fecal indicators is critical; however, changes in E. coli survival along the freshwater salinity range (≤ 1500 µS cm-1) have not been previously examined. Through a series of controlled mesocosm experiments, we provide direct evidence that salinization causes E. coli survival rates in freshwater to increase at conductivities as low as 350 µS cm-1 and peak at 1500 µS cm-1, revealing a subsidy-stress response across the freshwater-marine continuum. Furthermore, specific base cations affect E. coli survival differently, with Mg2+ increasing E. coli survival rates relative to other chloride salts. Further investigation of the mechanisms by which freshwater salinization increases susceptibility to or exacerbates bacterial water quality impairments is recommended. Addressing salinization with nuanced approaches that consider salt sources and chemistry could assist in prioritizing and addressing bacterial water quality management.

Cross-comparison of methods for quantifying antibiotic resistance in agricultural soils amended with dairy manure and compost.

Agricultural soils are often amended with livestock manure, making them a key reservoir of antibiotic resistance genes (ARGs). Given that soils are among the most microbially-diverse environments on the planet; effective characterization and quantification of the effects of manure-derived amendments on soil resistomes is a major challenge. This study examined the effects of dairy manure-derived amendments on agricultural soils via two strategies: quantification of anthropogenic ARG markers via qPCR and shotgun metagenomic resistome profiling; and these strategies were compared to a previously published antibiotic resistant fecal coliform dataset. Soil samples were collected throughout a 120 day complete block field experiment to compare the effects of amendment type on antibiotic resistance. Results of all three measurements were consistent with the hypothesis that the application of composted manure reduced antibiotic resistance in soil relative to the application of raw manure, although some differences were noted in comparing the patterns of the three measurements with time. Raw dairy manure-amended soils yielded high sul1 and tet(W) relative abundances on Day 0 (following amendment application), but significantly decreased to background levels by Day 67 (harvest) and Day 120 (study completion). Shotgun metagenomics similarly detected a decrease in the relative abundances of sulfonamide and tetracycline-associated ARGs over time in the raw manure- and compost-amended soils; however, these levels were significantly lower than those estimated by qPCR. Interestingly, although patterns of sulfonamide and tetracycline resistance among culturable fecal coliforms echoed those observed via qPCR and metagenomics; erythromycin resistant coliforms were directly recovered by culture in amended soils, but corresponding ARGs were not detected by qPCR or metagenomics. This study supports both composting and time restrictions as means of reducing the potential for antibiotic resistance in manure to spread via soil application. Results suggest some differences in finer conclusions drawn depending on which antibiotic resistance monitoring target is selected.

Making waves: Right in our backyard- surface discharge of untreated wastewater from homes in the United States.

Roughly » of U.S. residents (80 million people) lack access to sanitary sewers and are required to treat their wastewater through a permitted onsite wastewater treatment system (OWTS). The vast majority use conventional septic systems with subsurface infiltration, which work well under most conditions. However, certain geologic conditions (e.g., impermeable soil, high water table) can preclude use of septic systems, requiring investment in expensive advanced OWTS. The confluence of lack of sewer, unsuitable geology, and poverty can lead households to have no feasible option for treating wastewater. In many such communities households discharge raw sewage onto the ground through what are commonly called "straight pipes." Here, we present the first effort to synthesize available evidence documenting the scope of straight pipe use in the U.S., including estimates of close to 50% straight pipe use in some counties. Despite reports that straight pipes are widespread and troubling preliminary evidence of adverse health effects, there has been no national effort to estimate the use or impacts of straight pipes. There are various disincentives that discourage the reporting of straight pipes by both residents and government actors. We propose ways to improve quantification of straight pipes and increase knowledge of their adverse effects. We identify the characteristics of areas with large proportions of straight pipes and describe the role of new and pending government programs in encouraging reporting and providing solutions.

Fecal Indicator Bacteria and Antibiotic Resistance Genes in Storm Runoff from Dairy Manure and Compost-Amended Vegetable Plots.

Given the presence of antibiotics and resistant bacteria in livestock manures, it is important to identify the key pathways by which land-applied manure-derived soil amendments potentially spread resistance. The goal of this field-scale study was to identify the effects of different types of soil amendments (raw manure from cows treated with cephapirin and pirlimycin, compost from antibiotic-treated or antibiotic-free cows, or chemical fertilizer only) and crop type (lettuce [ L.] or radish [ L.]) on the transport of two antibiotic resistance genes (ARGs; 1 and ) via storm runoff from six naturally occurring storms. Concurrent quantification of sediment and fecal indicator bacteria (FIB; and enterococci) in runoff permitted comparison to traditional agricultural water quality targets that may be driving factors of ARG presence. Storm characteristics (total rainfall volume, storm duration, etc.) significantly influenced FIB concentration (two-way ANOVA, < 0.05), although both effects from individual storm events (Kruskal-Wallis, < 0.05) and vegetative cover influenced sediment levels. Composted and raw manure-amended plots both yielded significantly higher 1 and B levels in runoff for early storms, at least 8 wk following initial planting, relative to fertilizer-only or unamended barren plots. There was no significant difference between 1 or B levels in runoff from plots treated with compost derived from antibiotic-treated versus antibiotic-free dairy cattle. Our findings indicate that agricultural fields receiving manure-derived amendments release higher quantities of these two "indicator" ARGs in runoff, particularly during the early stages of the growing season, and that composting did not reduce effects of ARG loading in runoff.

Macroinvertebrate sensitivity thresholds for sediment in Virginia streams.

Sediment is the most commonly identified pollutant associated with macroinvertebrate community impairments in freshwater streams nationwide. Management of this physical stressor is complicated by the multiple measures of sediment available (e.g., suspended, dissolved, bedded) and the variability in natural "healthy" sediment loadings across ecoregions. Here we examine the relative importance of 9 sediment parameters on macroinvertebrate community health as measured by the Virginia Stream Condition Index (VSCI) across 5 ecoregions. In combination, sediment parameters explained 27.4% of variance in the VSCI in a multiregion data set and from 20.2% to 76.4% of variance for individual ecoregions. Bedded sediment parameters had a stronger influence on VSCI than did dissolved or suspended parameters in the multiregion assessment. However, assessments of individual ecoregions revealed conductivity had a key influence on VSCI in the Central Appalachian, Northern Piedmont and Piedmont ecoregions. In no case was a single sediment parameter sufficient to predict VSCI scores or individual biological metrics. Given the identification of embeddedness and conductivity as key parameters for predicting biological condition, we developed family-level sensitivity thresholds for these parameters, based on extirpation. Resulting thresholds for embeddedness were 68% for combined ecoregions, 65% for the Mountain bioregion (composed of Central Appalachian, Ridge and Valley, and Blue Ridge ecoregions), and 88% for the Piedmont bioregion (composed of Northern Piedmont and Piedmont ecoregions). Thresholds for conductivity were 366 µS/cm for combined ecoregions, 391 µS/cm for the Mountain bioregion, and 136 µS/cm for the Piedmont bioregion. These thresholds may help water quality professionals identify impaired and at-risk waters designated to support aquatic life and develop regional strategies to manage sediment-impaired streams. Inclusion of embeddedness as a restoration endpoint may be warranted; this could be facilitated by application of more quantitative, less time-intensive measurement approaches. We encourage refinement of thresholds as additional data and genus-based metrics become available. Integr Environ Assess Manag 2019;15:77-92. Published 2018. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Fate of Pirlimycin and Antibiotic-Resistant Fecal Coliforms in Field Plots Amended with Dairy Manure or Compost during Vegetable Cultivation.

Identification of agricultural practices that mitigate the environmental dissemination of antibiotics is a key need in reducing the prevalence of antibiotic-resistant bacteria of human health concern. Here, we aimed to compare the effects of crop (lettuce [ L.] or radish [ L.]), soil amendment type (inorganic fertilizer, raw dairy manure, composted dairy manure, or no amendment), and prior antibiotic use history (no antibiotics during previous lactation cycles vs. manure mixed from cows administered pirlimycin or cephapirin) of manure-derived amendments on the incidence of culturable antibiotic-resistant fecal coliforms in agricultural soils through a controlled field-plot experiment. Antibiotic-resistant culturable fecal coliforms were recoverable from soils across all treatments immediately after application, although persistence throughout the experiment varied by antibiotic class and time. The magnitude of observed coliform counts differed by soil amendment type. Compost-amended soils had the highest levels of cephalosporin-resistant fecal coliforms, regardless of whether the cows from which the manure was derived were administered antibiotics. Samples from control plots or those treated with inorganic fertilizer trended toward lower counts of resistant coliforms, although these differences were not statistically significant. No statistical differences were observed between soils that grew leafy (lettuce) versus rooted (radish) crops. Only pirlimycin was detectable past amendment application in raw manure-amended soils, dissipating 12 to 25% by Day 28. Consequently, no quantifiable correlations between coliform count and antibiotic magnitude could be identified. This study demonstrates that antibiotic-resistant fecal coliforms can become elevated in soils receiving manure-derived amendments, but that a variety of factors likely contribute to their long-term persistence under typical field conditions.

Engineered and Environmental Controls of Microbial Denitrification in Established Bioretention Cells.

Bioretention cells (BRCs) are effective tools for treating urban stormwater, but nitrogen removal by these systems is highly variable. Improvements in nitrogen removal are hampered by a lack of data directly quantifying the abundance or activity of denitrifying microorganisms in BRCs and how they are controlled by original BRC design characteristics. We analyzed denitrifiers in twenty-three BRCs of different designs across three mid-Atlantic states (MD, VA, and NC) by quantifying two bacterial denitrification genes ( nirK and nosZ) and potential enzymatic denitrification rates within the soil medium. Overall, we found that BRC design factors, rather than local environmental variables, had the greatest effects on variation in denitrifier abundance and activity. Specifically, denitrifying populations and denitrification potential increased with organic carbon and inorganic nitrogen concentrations in the soil media and decreased in BRCs planted with grass compared to other types of vegetation. Furthermore, the top layers of BRCs consistently contained greater concentrations and activity of denitrifying bacteria than bottom layers, despite longer periods of saturation and the presence of permanently saturated zones designed to promote denitrification at lower depths. These findings suggest that there is still considerable potential for design improvements when constructing BRCs that could increase denitrification and mitigate nitrogen export to receiving waters.

Movement of traditional fecal indicator bacteria and source-tracking targets through septic drainfields.

The past three decades' data on outbreaks in the United States indicate that homes dependent on untreated groundwater (e.g. wells) for household drinking water that are also reliant on onsite treatment of household wastewater (e.g. septic systems) may be at greater risk for waterborne disease. While groundwater quality monitoring to protect public health has traditionally focused on the detection of fecal indicator bacteria, the application of emerging source tracking strategies may offer a more efficient means to identify pollution sources and effective means of remediation. This study compares the movement of common fecal indicator bacteria (E. coli and enterococci) with a chemical (optical brighteners, OB) and a molecular (Bacteroides HF183) source tracking (ST) target in small scale septic drainfield models in order to evaluate their potential utility in groundwater monitoring. Nine PVC column drainfield models received synchronized doses of primary-treated wastewater twice daily, with influent and effluent monitored bi-weekly over a 7-month period for all targets. Results indicate that E. coli and enterococci concentrations were strongly associated (Spearman's rank, p<0.05), and correlations between enterococci and optical brighteners were moderately strong. Bacteroides HF183 was significantly, but not strongly, associated with optical brighteners and both indicator bacteria (Point-biserial correlation, p<0.05), most likely due to its sporadic detection. Application of human ST marker monitoring in groundwaters at risk of contamination by human sewage is recommended, although consistent interpretation of results will rely on more detailed evaluation of HF183 incidence in source contamination waters.

Interactive effects of water quality, physical habitat, and watershed anthropogenic activities on stream ecosystem health.

Ecological degradation of streams remains a major environmental concern worldwide. While stream restoration has received considerable attention, mitigation efforts focused on the improvement of physical habitat have not proven completely effective. Several small-scale studies have emphasized that effective restoration strategies require a more holistic understanding of the variables at play, although the generalization of the findings based on the small-scale studies remains unclear. Using a comprehensive statewide stream monitoring database from West Virginia (WV), a detailed landscape dataset, and a machine learning algorithm, this study explores the interactive impacts of water quality and physical habitat on stream ecosystem health as indicated by benthic macroinvertebrate scores. Given the long history of energy extraction in this region (i.e., coal mining and oil/gas production), investigation of energy extraction influences is highlighted. Our results demonstrate that a combination of good habitat and low specific conductance is generally associated with favorable benthic macroinvertebrate scores, whereas poor habitat combined with water quality conditions typically indicative of high ionic strength are associated with impaired stream status. In addition, streams impacted by both energy extraction and residential development had a higher percentage of impairment compared to those impacted predominantly by energy extraction or residential development alone. While water quality played a more important role in the ecosystem health of streams impacted primarily by energy extraction activities, habitat seems to be more influential in streams impacted by residential development. Together, these findings emphasize that stream restoration strategies should consider interactive effects of multiple environmental stressors tailored to specific sites or site types - as opposed to considering a single stressor or multiple stressors separately.

Tracking the downstream impacts of inadequate sanitation in central Appalachia.

Poor sanitation in rural infrastructure is often associated with high levels of fecal contamination in adjacent surface waters, which presents a community health risk. Although microbial source tracking techniques have been widely applied to identify primary remediation needs in urban and/or recreational waters, use of human-specific markers has been more limited in rural watersheds. This study quantified the human source tracking marker Bacteroides-HF183, along with more general fecal indicators (i.e. culturable Escherichia coli and a molecular Enterococcus marker), in two Appalachian streams above and below known discharges of untreated household waste. Although E. coli and Enterococcus were consistently recovered in samples collected from both streams, Bacteroides-HF183 was only detected sporadically in one stream. Multiple linear regression analysis demonstrated a positive correlation between the concentration of E. coli and the proximity and number of known waste discharge points upstream; this correlation was not significant with respect to Bacteroides-HF183, likely due to the low number of quantifiable samples. These findings suggest that, while the application of more advanced source targeting strategies can be useful in confirming the influence of substandard sanitation on surface waters to justify infrastructure improvements, they may be of limited use without concurrent traditional monitoring targets and on-the-ground sanitation surveys.