Wastewater Treatment Plant Outfall Research Articles

Microbial community structure and diversity attached to the periphyton in different urban aquatic habitats.

Periphyton is a complex community composed of diverse prokaryotes and eukaryotes; understanding the characteristics of microbial communities within periphyton becomes crucial for biogeochemical cycles and energy dynamics of aquatic ecosystems. To further elucidate the community characteristics of periphyton across varied aquatic habitats, including unpolluted ecologically restored lakes, aquaculture ponds, and areas adjacent to domestic and industrial wastewater treatment plant outfalls, we explored the composition and diversity of prokaryotic and eukaryotic communities in periphyton by employing Illumina MiSeq sequencing. Our findings indicated that the prokaryotic communities were predominantly composed of Proteobacteria (40.92%), Bacteroidota (21.01%), and Cyanobacteria (10.12%), whereas the eukaryotic communities were primarily characterized by the dominance of Bacillariophyta (24.09%), Chlorophyta (20.83%), and Annelida (15.31%). Notably, Flavobacterium emerged as a widely distributed genus among the prokaryotic community. Unclassified_Tobrilidae exhibited higher abundance in unpolluted ecologically restored lakes. Chaetogaster and Nais were enriched in aquaculture ponds and domestic wastewater treatment plant outfall area, respectively, while Surirella and Gomphonema dominated industrial sewage treatment plant outfall area. The alpha diversity of eukaryotes was higher in unpolluted ecologically restored lakes. pH and nitrogen content ( , , and TN) significantly explained the variations for prokaryotic and eukaryotic community structures, respectively. Eukaryotic communities exhibited a more pronounced response to habitat variations compared to prokaryotic communities. Moreover, the association networks revealed an intensive positive correlation between dominant Bacillariophyta and Bacteroidota. This study provided useful data for identifying keystone species and understanding their ecological functions.

Microparticles in Wild and Caged Biota, Sediments, and Water Relative to Large Municipal Wastewater Treatment Plant Discharges.

Anthropogenically modified microparticles including microplastics are present in municipal wastewater treatment plant (WWTP) effluents; however, it is unclear whether biotic exposures are elevated downstream of these outfalls. In the fall of 2019, the present study examined whether microparticle levels in resident fish, environmental samples, and caged organisms were elevated near the Waterloo and Kitchener WWTP outfalls along the Grand River, Ontario, Canada. Wild rainbow darters (Etheostoma caeruleum) were collected from a total of 10 sites upstream and downstream of both WWTPs, along with surface water and sediment samples to assess spatial patterns over an approximately 70-km river stretch. Amphipods (Hyalella azteca), fluted-shell mussels (Lasmigona costata), and rainbow trout (Oncorhynchus mykiss) were also caged upstream and downstream of one WWTP for 14 or 28 days. Whole amphipods, fish digestive tracts, and mussel tissues (hemolymph, digestive glands, gills) were digested with potassium hydroxide, whereas environmental samples were processed using filtration and density separation. Visual identification, measurement, and chemical confirmation (subset only) of microparticles were completed. Elevated abiotic microparticles were found at several upstream reference sites as well as at one or both wastewater-impacted sites. Microparticles in amphipods, all mussel tissues, and wild fish did not show patterns indicative of increased exposures downstream of effluent discharges. In contrast, elevated microparticle counts were found in trout caged directly downstream of the outfall. Across all samples, cellulose fibers (mainly blue and clear colors) were the most common. Overall, results suggest little influence of WWTP effluents on microparticles in biota but rather a ubiquitous presence across most sites that indicates the importance of other point and nonpoint sources to this system. Environ Toxicol Chem 2024;43:1047-1061. © 2024 His Majesty the King in Right of Canada and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Environmental monitoring of antimicrobial resistant bacteria in North Carolina water and wastewater using the WHO Tricycle protocol in combination with membrane filtration and compartment bag test methods for detecting and quantifying ESBL E. coli

Antimicrobial resistance (AMR) threatens human and animal health; effective response requires monitoring AMR presence in humans, animals, and the environment. The World Health Organization Tricycle Protocol (WHO TP) standardizes and streamlines global AMR monitoring around a single indicator organism, extended-spectrum-β-lactamase-producing Escherichia coli (ESBL-Ec). The WHO TP culture-based method detects and quantifies ESBL-Ec by spread-plating or membrane filtration on either MacConkey or TBX agar (supplemented with cefotaxime). These methods require laboratories and trained personnel, limiting feasibility in low-resource and field settings. We adapted the WHO TP using a simplified method, the compartment bag test (CBT), to quantify most probable numbers (MPN) of ESBL-Ec in samples. CBT methods can be used correctly in the field by typical adults after a few hours’ training. We collected and analyzed municipal wastewater, surface water, and chicken waste samples from sites in Raleigh and Chapel Hill, NC over an 8-month period. Presumptive ESBL-Ec were quantified using MF on TBX agar supplemented with cefotaxime (MF+TBX), as well as using the CBT with chromogenic E. coli medium containing cefotaxime. Presumptive ESBL-Ec bacteria were isolated from completed tests for confirmation and characterization by Kirby Bauer disk diffusion tests (antibiotic sensitivity) and EnteroPluri biochemical tests (speciation). Both methods were easy to use, but MF+TBX required additional time and effort. The proportion of E. coli that were presumptively ESBL in surface water samples was significantly greater downstream vs upstream of wastewater treatment plant (WWTP) outfalls, suggesting that treated wastewater is a source of ESBL-Ec in some surface waters. The CBT and MF+TBX tests provided similar (but not identical) quantitative results, making the former method suitable as an alternative to the more complex MF+TBX procedure in some applications. Further AMR surveillance using MF+TBX and/or CBT methods may be useful to characterize and refine their performance for AMR monitoring in NC and elsewhere.

Effects of droughts, sea level rise, and increase in outfall discharges on phytoplankton in a temperate estuary (Tagus Estuary, Portugal)

The effects of climate change on the estuarine environment are not fully understood. In temperate regions, droughts are expected to increase in frequency and severity, due to lower precipitation, and the mean sea level is expected to rise. This study aimed to assess how the estuarine environment will be affected by river flow’s reduction, mean sea level rise, and the increase in nutrients discharged from anthropogenic sources. Seven scenarios were simulated and analyzed in the Tagus estuary (Portugal), using the hydrodynamic and biogeochemical model SCHISM: (i) reference scenario, (ii) 10% increase of the wastewater treatment plant (WWTP) outfall’s discharge, (iii) 25% reduction of the river flow, (iv) 50% reduction of the river flow, (v) sea level rise of 0.5 m, (vi) sea level rise of 1 m, and (vii) the combination of 0.5 m of sea level rise and 25% reduction of the river flow. Both the reduction of the river flow and mean sea level rise led to higher salinities and lower nutrients and chlorophyll-a concentrations in the mid and upper areas of the estuary. The reduction in riverine nutrients in the estuary may increase the importance of nutrients from anthropogenic sources (e.g. WTTP discharges) in shaping the spatial variability of the phytoplankton communities in the future.

Changes in chemical occurrence, concentration, and bioactivity in the Colorado River before and after replacement of the Moab, Utah wastewater treatment plant

Long-term (2010–19) water-quality monitoring on the Colorado River downstream from Moab Utah indicated the persistent presence of Bioactive Chemicals (BC), such as pesticides and pharmaceuticals. This stream reach near Canyonlands National Park provides critical habitat for federally endangered species. The Moab wastewater treatment plant (WWTP) outfall discharges to the Colorado River and is the nearest potential point-source to this reach. The original WWTP was replaced in 2018. In 2016–19, a study was completed to determine if the new plant reduced BC input to the Colorado River at, and downstream from, the outfall. Water samples were collected before and after the plant replacement at sites upstream and downstream from the outfall. Samples were analyzed for as many as 243 pesticides, 109 pharmaceuticals, 20 hormones, 51 wastewater indicator chemicals, 20 metals, and 8 nutrients. BC concentrations, hazard quotients (HQs), and exposure activity ratios (EARs) were used to identify and prioritize contaminants for their potential to have adverse biological effects on the health of native and endangered wildlife. There were 22 BC with HQs >1, mostly metals and hormones; and 23 BC with EARs >0.1, mostly hormones and pharmaceuticals. Most high HQs or EARs were associated with samples collected at the WWTP outfall site prior to its replacement. Discharge from the new plant had reduced concentrations of nutrients, hormones, pharmaceuticals, and other BC. For example, all 16 of the hormones detected at the WWTP outfall site had maximum concentrations in samples collected prior to the WWTP replacement. The WWTP replacement had less effect on instream concentrations of metals and pesticides, BC whose sources are less directly tied to domestic wastewater. Study results indicate that improved WWTP technology can create substantial reductions in concentrations of non-regulated BC such as pharmaceuticals, in addition to regulated contaminants such as nutrients.

Spatial distribution and temporal trends of cyclic and linear siloxanes in sediment from semi-enclosed and industrialized bays of Korea, in 2013 and 2021

Although siloxanes are contaminants of emerging concerns, limited studies have been conducted on contamination and time trends in siloxanes from aquatic environments worldwide. To date, most previous studies have focused on the environmental relevance of cyclic siloxanes in coastal environments. In the present study, cyclic and linear siloxanes were measured in sediment collected from semi-enclosed bays in Korea in 2013 and 2021 to assess occurrence, spatial distribution, temporal trends, and ecological risks. Almost all siloxanes were detected in all sediment samples, indicating continuous contamination for the last decade. The concentrations of cyclic siloxanes in sediment were approximately two times higher than those of linear siloxanes. Decamethylcyclopentasiloxane (D5) and dodecamethylcyclohexasiloxane (D6) were predominantly detected in all sediment samples, possibly due to their high consumption and strong resistance. The overall concentrations of siloxanes in sediment gradually decreased from inner to outer parts of the bays, suggesting that industrial activities largely contributed to siloxane contamination. Moreover, the highest concentrations of siloxanes in both sampling years were observed in sediment near the outfall of wastewater treatment plant, implying a potential source in the coastal environment. Siloxane concentrations in sediment were significantly correlated with sedimentary organic carbon. The concentrations of siloxanes in sediment were not significantly different between 2013 and 2021, indicating ongoing contamination. The concentrations of D5 and D6 in our sediment samples showed a limited potential to pose a threat to benthic organisms.

Chemical prioritization of pharmaceuticals and personal care products in an urban tributary of the Potomac River

Pharmaceuticals and personal care products (PPCPs) are incredibly diverse in terms of chemical structures, physicochemical properties, and modes of action, making their environmental impacts challenging to assess. New chemical prioritization methodologies have emerged that compare contaminant monitoring concentrations to multiple toxicity data sources, including whole organism and high-throughput data, to develop a list of “high priority” chemicals requiring further study. We applied such an approach to assess PPCPs in Hunting Creek, an urban tributary of the Potomac River near Washington, DC, which has experienced extensive human population growth. We estimated potential risks of 99 PPCPs from surface water and sediment collected upstream and downstream of a major wastewater treatment plant (WWTP), nearby combined sewer overflows (CSO), and in the adjacent Potomac River. The greatest potential risks to the aquatic ecosystem occurred near WWTP and CSO outfalls, but risk levels rapidly dropped below thresholds of concern – established by previous chemical prioritization studies – in the Potomac mainstem. These results suggest that urban tributaries, rather than larger rivers, are important to monitor because their lower or intermittent flow may not adequately dilute contaminants of concern. Common psychotropics, such as fluoxetine and venlafaxine, presented the highest potential risks, with toxicity quotients often > 10 in surface water and > 1000 in sediment, indicating the need for further field studies. Several ubiquitous chemicals such as caffeine and carbamazepine also exceeded thresholds of concern throughout our study area and point to specific neurotoxic and endocrine modes of action that warrant further investigation. Since many “high priority” chemicals in our analysis have also triggered concerns in other areas around the world, better coordination is needed among environmental monitoring programs to improve global chemical prioritization efforts.

The influence of environmental and ecological factors on the accumulation and distribution of short- and long-chain perfluoroalkyl acids in a mid-trophic avian insectivore

Perfluoroalkyl acids (PFAAs) include perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonic acid (PFHxS), and perfluorodecane sulfonic acid (PFDS), as well as increasingly used alternative short-chain perfluorosulfonic acids (PFSAs) and short- and long-chain (≥C9) perfluorocarboxylic acids (PFCAs). In the present study, tissues of tree swallows (Tachycineta bicolor) from two sites in southern Ontario, Canada, were analyzed for 17 individual PFAAs and showed egg and nestling tissue (liver, carcass) profiles dominated by PFOS (57–66%). The remaining PFAAs contributed ≤7% each, although collectively the long-chain PFCAs comprised 21–29% of the PFAAs. The short-chain PFSAs and PFCAs were among the lowest concentrations, suggesting that despite increased production and use of these alternative PFAAs, they are not accumulated to the same extent as the long-chain PFSAs and PFCAs. PFOS, PFDS, and some long-chain PFCAs were significantly higher in eggs than in livers and carcasses, whereas PFOA and the two short-chain PFCAs were significantly higher in nestling tissues than in eggs. For the two short-chain PFSAs, concentrations were similar among tissues. Tree swallow tissues at the site near a wastewater treatment plant (WWTP) outfall showed higher concentrations of PFOS, PFDS, PFHxS, and some long-chain PFCAs than tree swallows sampled at the nearby reference site; however, the influence of the WWTP was more equivocal for PFOA, other long-chain PFCAs, and short-chain PFSAs and PFCAs. Carbon stable isotopes (δ13C) and fatty acid signatures indicated that the diets of the WWTP swallows were more terrestrial than the reference swallows. Nonetheless, models considering environmental and ecological variables indicated that site was often the primary driver of PFAA variation among the swallows, with less or no influence of dietary patterns, or sex or body condition, revealing that of WWTP effluent can be an important environmental source of the major PFAAs in tree swallows.

Pharmaceutical pollution in marine waters and benthic flora of the southern Australian coastline

Environmental context Most human pharmaceutical waste is discharged to the environment. While the presence of pharmaceuticals in freshwater systems is well documented globally, little is known of the impact on marine ecosystems. We measured pharmaceuticals in a marine environment in south-eastern Australia and found pharmaceutical concentrations around 24 000 times higher in benthic flora than in the marine surface waters. We discuss the potential use of seaweeds as biological indicators of pharmaceutical pollution. Rationale Pharmaceuticals are emerging pollutants of concern with a range of adverse consequences for organisms and ecosystems. Their presence in freshwater and estuarine systems has been well documented, but less is known about their prevalence in open ocean, or their uptake by benthic flora. This preliminary survey of the southern Australian coastline sought to measure the concentrations of key pharmaceuticals in both surface waters and benthic flora. Methodology This study used LC-MS/MS to measure the concentration carbamazepine, tramadol and venlafaxine in (1) samples from wastewater treatment plants, (2) ocean surface waters and (3) several species of benthic flora. Surface waters and benthic flora were sampled at two sites near waste water treatment plant (WWTP) discharges, and one site away from any discharge. Results All three pharmaceuticals were detected in surface water samples with their risk assessed (via risk quotient) as medium risk (carbamazepine) or low risk (venlafaxine, tramadol). All three pharmaceuticals were also detected in benthic flora, particularly in brown macroalgae; Tramadol was measured at a maximum of 34.7 ng g−1 in Hormosira banksii, and Venlafaxine was recorded at a maximum of 17.3 ng g−1 in Caulocystis cephalornithos. Discussion The calculated bioconcentration factors suggest the pharmaceutical concentrations in benthic flora were up to ~24 000 times higher than in surrounding surface water. There was also evidence that proximity to WWTP outfalls influenced the levels of pharmaceuticals in benthic flora. The results suggest that the benthic flora may be suitable bioindicators of pharmaceutical contamination and that the potential impacts of pharmaceutical pollutants in marine ecosystems demand further investigation.

Wild fish responses to wastewater treatment plant upgrades in the Grand River, Ontario

Municipal wastewater treatment plant (WWTP) effluent is one of several point sources of contaminants (nutrients, pharmaceuticals, estrogens, etc.) which can lead to adverse responses in aquatic life. Studies of WWTP effluent impacts on rainbow darter (Etheostoma caeruleum) collected downstream of WWTPs in the Grand River, Ontario have reported disruption at multiple levels of biological organization, including altered vitellogenin gene expression, lower levels of in vitro steroid production, and high frequency of intersex. However, major upgrades have occurred at treatment plants in the central Grand River over the last decade. Treatment upgrades to the Waterloo WWTP were initiated in 2009 but due to construction delays, the upgrades came fully on-line in 2017/2018. Responses in rainbow darter have been followed at sites associated with the outfall consistently over this entire time period. The treatment plant upgrade resulted in nitrification of effluent, and once complete there was a major reduction in effluent ammonia, selected pharmaceuticals, and estrogenicity. This study compared several key responses in rainbow darter associated with the Waterloo WWTP outfall prior to and post upgrades. Stable isotopes signatures in fish were used to track exposure to effluent and changed dramatically over time, corresponding to the effluent quality. Disruptions in in vitro steroid production and intersex in the darters that had been identified prior to the upgrades were no longer statistically different from the upstream reference sites after the upgrades. Although annual variations in water temperature and flow can potentially mask or exacerbate the effects of the WWTP effluent, major capital investments in wastewater treatment targeted at improving effluent quality have corresponded with the reduction of adverse responses in fish in the receiving environment.

Spatial Variability of Physicochemical Parameters and Phytoplankton at the Tagus Estuary (Portugal)

The estuarine phytoplankton communities are known to respond rapidly to environmental changes, being considered an important water quality indicator; thus, it is crucial to fully understand its natural variability. The objective of the present study was to assess the spatial variability of both physicochemical variables and the phytoplankton community, to understand how such variability is influenced by seasonality and to evaluate how the anthropogenic sources affect such patterns. The Tagus estuary was used as a case study, since it is one of Europe’s largest estuaries, with high spatial and seasonal variations and a high level of human pressure associated with large urban and industrial areas. To achieve this goal, environmental parameters, nutrients concentration, bivalve biomass (filter feeders) and phytoplankton pigments were quantified in a single summer sampling campaign with high spatial resolution and in monthly campaigns in eight sampling stations through the Tagus estuary, in one year. In general, suspended particulate matter and nutrients decreased from the upper part of the estuary to the estuary mouth; however, relevant local inputs were also observed in more downstream locations, near outfalls of wastewater treatment plants (WWTP). The chlorophyll-a concentrations were higher over the southern intertidal mudflats, probably due to resuspension of microphytobenthos, associated with higher nutrient concentrations. Through a grazing indicator, it was observed that grazers are important drivers of variability of the phytoplankton community composition. All water bodies achieved “good” and “high” water quality classifications for both physicochemical and biological indicators, with the worst results reported for the water bodies located at the upper estuary. Therefore, this estuary presents a decreasing trend of nutrients and chlorophyll-a in the upstream–downstream direction, except for the estuary channels and the outfall in the northern margin, which lead to an increase in nutrient concentrations. However, these increases did not affect the water quality of the three analyzed water bodies, presenting at least good ecological status, considering the nutrient and chlorophyll-a indicators.

Spatial and temporal variations in anti-androgenic activity and environmental risk in a small river

The biological effects of multiple compounds have been widely investigated in aquatic environments. However, investigations of spatial and temporal variations in biological effects are rarely performed because they are time-consuming and labor-intensive. In this study, the variability of the anti-androgen, receptor-mediated activity of surface water samples was observed over 3 years using in vitro bioassays. Large-volume water samples were collected at one site upstream (Wer site) and two sites downstream (Sil and Nien sites) of a wastewater treatment plant (WWTP) outfall in the Holtemme River. Anti-AR activity was persistently present in all surface water samples over the three years. Large spatial variations in anti-androgenic activity were observed, with the lowest activity at the Wer site (mean concentration of 9.5 ± 7.2 μg flutamide equivalents/L) and the highest activity at the Sil site (mean concentration of 31.1 ± 12.0 μg flutamide equivalents/L) directly influenced by WWTP effluents. On the temporal scale, no distinct trend for anti-AR activity was observed among the seasons in all three years. The anti-androgenic activity at the upstream Wer site showed a decreasing trend from 2014 to 2016, indicating improved water quality. A novel bioanalytical-equivalent-based risk assessment method considering the frequency of risk occurrence was developed and then utilized to assess the environmental risk of anti-androgenic activity in the Holtemme River. The results revealed that the highest risk was present at the Sil site, while the risk was considerably reduced at the Nien site. The risk at the upstream Wer site was the lowest.

Assessing multiple fecal sources to surf zone waters of two recreational beaches by bacterial community analysis

Fecal sources to recreational surf zone waters should be identified to protect public health. While watershed origins of human and other fecal sources are often discoverable by quantitative polymerase chain reaction (qPCR) of fecal markers using spatially stratified samples, similarly assessing wastewater treatment plant (WWTP) outfall and other offshore contributions to surf zones is challenged by individual marker fate and transport. Here, bacterial communities were assessed for relatedness between all hypothesized fecal sources and surf zone waters for two urban California recreational beaches, by sequencing genes encoding 16S rRNA and analyzing data using SourceTracker and FEAST. Ambient marine bacterial communities dominated the surf zone, while fecal (human, dog, or gull) or wastewater (sewage or treated WWTP effluent) bacterial communities were present at low proportions and those from recycled water were absent. Based on the relative abundances of bacterial genera specifically associated with human feces, the abundances of HF183 in bacterial community sequences, and FEAST and SourceTracker results when benchmarked to HF183, the major sources of HF183 to surf zone waters were human feces and treated WWTP effluent. While surf zone sequence proportions from human sources (feces, sewage and treated WWTP effluent) appeared uncorrelated to previously obtained qPCR HF183 results, the proportions of human fecal and potential human pathogen sequences in surf zone waters were elevated when there were more swimmers (i.e. during weekday afternoons, holidays and busy weekends, and race events), thus confirming previously-published qPCR-based conclusions that bather shedding contributed low levels of human fecal contamination. Here, bacterial community sequencing also showed evidence that treated WWTP effluent from an offshore outfall was entering the surf zone, thereby resolving a prior uncertainty. Thus, bacterial community sequencing not only confirms qPCR HF183-based human marker detections, but further allows for confirming fecal sources for which individual marker quantification results can be equivocal.

Environmental impact assessment of discharge of treated wastewater effluent in Upper Iskar sub-catchment

The upper Iskar sub-catchment is one of Bulgaria’s most important economic and socially significant water sources because of its role in supplying Sofia with drinking water. Among the critical factors that carry potential high-risk levels for water quality in this hydrosystem are the discharge from the Samokov Wastewater Treatment Plant (WWTP), diffuse pollution from agriculture, and the percolation of untreated sewage from the small villages. In this study, we assessed the effect of treated wastewater effluent on water quality, and on the ecological state and microbial communities in the river sector of Samokov’s WWTP discharge area. The assessment was based on the complex use of chemical and microbiological indicators and biological quality elements. The concentrations of organics, nutrients and microcomponents were determined with results confirming the expected increase for parameters associated with the discharge of urban wastewater. The ecological state, according to macrozoobenthos indicators, was “good” throughout the river sector but local deterioration was registered in a proximal location downstream of the WWTP outfall. The analysis of stream water and bed sediment microbial communities by a fluorescent technique showed the high metabolic activity and intensive transformation processes in addition to high abundance registered with standard cultivation methods. The importance of the studied sub-catchment for the functioning of the urban water cycle, and for the quality of Sofia’s drinking water, underlines the need to extend an existing monitoring program with a more detailed assessment of the environmental impact of wastewater discharge.

Multiple Indicators of Wastewater Contamination to Shellfish Farms Near a Tidal River

Wastewater contamination threatens the shellfish aquaculture industry by posing risks to public health. Multiple indicators of wastewater contamination, including fecal coliforms (fc), male-specific coliphage (MSC), dissolved nutrients, stable isotope ratios, and artificial sweeteners were analyzed to determine possible sources of wastewater to local shellfish farms. Samples were collected at a wastewater treatment plant outfall (WTPO), nonpoint residential, and agricultural areas of a tidal river, and tidal creek inflows adjacent to farms. To capture seasonal variation, we sampled under warm and cold, and wet and dry conditions. Fc ranged < 5–5250 CFU 100 mL−1, NH4+ concentrations ranged up to 9.58 μM, and δ15N ranged 1.4–7.8‰ across all sites and time periods. Fc and NH4+ were higher, and δ15N was lower in the cold wet period and near residential and agricultural areas. Acesulfame and sucralose concentrations ranged 0.004–0.05 μg L−1 and up to > 0.8 μg L−1, respectively, and did not correlate with other indicators but tended to be higher in residential areas and at the WTPO, supporting their value in differentiating human sewage from other sources. Shoreline disturbance during septic system upgrades may have inadvertently contributed bacterial indicators to shellfish farms. Overall, indicator source dominance depended on environmental conditions, with WTPO and residential sources conveying human-specific indicators to farms year-round, while agricultural and industrial sites contributed additional fc during cold wet periods. The use of multiple indicators will aid managers to detect and define wastewater sources, identify targets for monitoring or remediation, and manage shellfish areas in estuaries with a mosaic of land-derived wastewater sources.

Dissolved rubidium to strontium ratio as a conservative tracer for wastewater effluent-sourced contaminant inputs near a major urban wastewater treatment plant

Municipal wastewater (MWW) effluent discharges can introduce contaminants to receiving waters which may have adverse impacts on local ecosystems and human health. Conservative chemical constituents specific to the MWW effluent stream can be used to quantify and trace wastewater effluent-sourced contaminant inputs. Gadolinium (Gd), a rare earth element used as a contrasting agent in medical magnetic resonance imaging, can be found in urban MWW streams. Dissolved anthropogenic Gd has been shown to be an indicator and potential conservative tracer for MWW effluent in receiving waters. Like other known MWW tracers, it can be difficult and expensive to measure. Dissolved rubidium (Rb) to strontium (Sr) ratio enrichment in biological materials such as blood and urine can lead to enriched Rb/Sr values in MWW effluent relative to natural waters. This ratio is relatively easy and inexpensive to measure and represents a promising additional indicator for MWW effluent in receiving waters in urbanized freshwater systems. In July 2015 and 2016 surface water samples were collected from sites in the tidal-fresh Potomac River in the vicinity of the Blue Plains Advanced Wastewater Treatment Plant (BPAWWTP) outfall near Washington, DC USA along with treated MWW effluent samples from the BPAWWTP. Dissolved Rb/Sr ratios were measured in these waters and compared to dissolved Gd concentrations in order to demonstrate the potential of the dissolved Rb/Sr ratio as a conservative indicator for MWW effluent. Results suggest the dissolved Rb/Sr ratio represents a simple and cost-effective indicator and conservative tracer for MWW effluent. It can be used with, or in place of, other proven tracers to investigate wastewater impacts in highly-urbanized, anthropogenically-impacted freshwater systems like the tidal fresh Potomac River and perhaps in a wider range of geologic settings than previously thought. A case study is presented as an example to demonstrate the potential of using dissolved Rb/Sr ratios to trace MWW-sourced nutrient inputs from a major WWTP like BPAWWTP to the receiving waters of tidal-fresh Potomac River.

Sources of Low Level Human Fecal Markers in Recreational Waters of Two Santa Barbara, CA Beaches: Roles of WWTP Outfalls and Swimmers

Worldwide, fecal indicator bacteria (FIB) evidence coastal water contamination for which sources are unknown. Here, for two FIB-impacted Santa Barbara recreational beaches, hypothesized fecal sources were investigated over three dry seasons (summers) using nearly 2000 field samples of water (ocean, creek, groundwater), sand, sediments, effluent and fecal sources. In years 1 and 2, gull and dog feces were identified as the probable main FIB sources to surf zone waters, yet HF183 human fecal markers were consistently detected. Determining HF183 sources was therefore prioritized, via year 3 sub-studies. In lower watersheds, human and dog wastes were mobilized by small storms into creeks, but no storm drain outfalls or creeks discharged into surf zones. Beach area bathrooms, sewers, and a septic system were not sources: dye tracing discounted hydraulic connections, and shallow groundwater was uncontaminated. Sediments from coastal creeks and downstream scour ponds, nearshore marine sediments, and sands from inter- and supratidal zones contained neither HF183 nor pathogens. Two nearby wastewater treatment plant (WWTP) outfalls discharged HF183 into plumes that were either deep or distant with uncertain onshore transport. Regardless, local sources were evidenced, as surf zone HF183 detection rates mostly exceeded those offshore and nearshore (around boat anchorages). The presence of swimmers was associated with surf zone HF183, as swimmer counts (on weekdays, holidays, weekends, and during races) significantly correlated (p<0.05, n = 196) to HF183 detections. Besides comprehensively assessing all possible fecal sources, this study provides new explanations of chronic low-level human markers in recreational beach surf zones, suggesting likely lowest achievable HF183 thresholds.

Pharmaceuticals and alkylphenols in transplanted Pacific oysters (Crassostrea gigas): Spatial variation and growth effects

Pharmaceuticals and personal care products in wastewater discharge can be stressors to estuarine species. We transplanted juvenile Pacific oysters at varying distances within sites near wastewater treatment plant outfalls or oyster aquaculture control sites to assess small scale spatial variation in contaminant uptake and oyster condition. Oysters were transplanted to sites in Coos and Netarts Bays, Oregon and Grays Harbor, Washington, then collected after 9 and 12 months. Two pharmaceuticals (miconazole and virginiamycin M1) were detected in spring samples and four alkylphenols (NP1EO, NP2EO, NP and OP) were detected in summer samples, with more frequent detections at wastewater sites. Contaminant concentrations were similar across site types, indicating that even in sparsely populated coastal areas (<25,000 in the watershed), shellfish are exposed to and uptake wastewater contaminants. Additionally, oyster condition was lower at wastewater sites compared to aquaculture sites, indicating a need to better understand whether contaminant exposure affects oyster condition.

Contrasting Raz–Rru stream metabolism and nutrient uptake downstream of urban wastewater effluent sites

Understanding how key stream ecosystem functions respond to wastewater treatment plant effluent is critical for assessing the ability of stream ecosystems to ameliorate anthropogenic nutrient loading and to effectively manage and restore impacted systems. We evaluated instream metabolism, reactive solute transport, and nutrient uptake along two 1.5 to 2-km-long reaches of a 2nd-order stream in the urbanized suburbs of Philadelphia, Pennsylvania, USA, each directly downstream of a wastewater treatment plant outfall. We compared metabolism based on resazurin–resorufin (Raz–Rru) with nutrient uptake and dissolved oxygen (DO) metabolism calculations. Plateau co-injections of the Raz–Rru metabolic tracer system and fluorescein provided integrated stream metabolism measurement. We sampled tracer concentrations hourly along longitudinal profiles and recorded them continuously, along with DO, at 2 discrete locations. The smaller reach 1, characterized by higher nutrient concentrations and canopy cover, had higher short-term transient storage and Raz uptake velocity. In contrast reach 2, with lower nutrient concentrations and less canopy cover, had higher nitrate and phosphorus uptake along with higher rates of gross primary productivity (GPP) and ecosystem respiration (ER). Temporal analysis indicated nitrate uptake increased over the afternoon at reach 2, whereas Raz uptake declined at both reaches. Our results suggest that nutrient uptake and GPP are sensitive to excessive nutrient concentrations and light in our system. In contrast, lower light and higher transient storage are likely driving larger, reach-scale spatial differences in Raz-based ER. Increasing nitrate uptake at reach 2, which lags behind diel DO concentrations, is likely the result of assimilatory N uptake coupled to GPP moderated by nitrification and denitrification, whereas decreasing rates of Raz transformation are likely related to diel variation in heterotrophic uptake. Our ability to resolve sub-daily changes in ER illustrates one of the key advantages of the Raz–Rru tracer system. However, our results also show the need for further investigation into the drivers of sub-daily ecosystem metabolism in streams as well as the mechanistic differences between DO- and Raz-based estimates of ER. Contrasting results from different measures of metabolic activity between reaches and over time highlight the complexity of metabolic processes in high-nutrient systems.

Micropollutant biotransformation and bioaccumulation in natural stream biofilms

Micropollutants are ubiquitously found in natural surface waters and pose a potential risk to aquatic organisms. Stream biofilms, consisting of bacteria, algae and other microorganisms potentially contribute to bioremediating aquatic environments by biotransforming xenobiotic substances. When investigating the potential of stream biofilms to remove micropollutants from the water column, it is important to distinguish between different fate processes, such as biotransformation, passive sorption and active bioaccumulation. However, due to the complex nature of the biofilm community and its extracellular matrix, this task is often difficult. In this study, we combined biotransformation experiments involving natural stream biofilms collected up- and downstream of wastewater treatment plant outfalls with the QuEChERS extraction method to distinguish between the different fate processes. The QuEChERS extraction proved to be a suitable method for a broad range of micropollutants (> 80% of the investigated compounds). We found that 31 out of 63 compounds were biotransformed by the biofilms, with the majority being substitution-type biotransformations, and that downstream biofilms have an increased biotransformation potential towards specific wastewater-relevant micropollutants. Overall, using the experimental and analytical strategy developed, stream biofilms were demonstrated to have a broad inherent micropollutant biotransformation potential, and to thus contribute to bioremediation and improving ecosystem health.