Diagnostic screening of private well water using gas chromatography with high resolution mass spectrometry to support well water management.

In the US, private well users are responsible for their own water quality testing, but local health officials are often uncertain what tests to recommend, particularly for regulated organic chemical contaminants. This study evaluated the utility of suspect and non-target screening (NTS) high resolution mass spectrometry (HRMS) as a tool to identify a wide range of organic chemicals of emerging concern in private well water and to inform well water management decisions. Qualitative NTS, which detects chemicals without providing concentrations, was used to analyze 25 private well water samples from Wake County, North Carolina. Using the NIST 20 mass spectral database (M1), NTS tentatively identified 106 unique organic chemicals across the 25 samples and an average of 11 organic chemicals per sample. At least one USEPA ToxCast chemical was identified in each private well water sample. Private well water users were interviewed prior to and after their sample's NTS results were reported back; four county groundwater managers were interviewed after aggregated results for all 25 water samples were reported back. All but one well user participant chose to participate in the reporting-back post-interview. The 24 private well users found NTS results useful and valued the contextualization of their results using NTS results for other well users and a local municipal water sample. Most private well users (67%) were surprised by their well water results, especially regarding the number of tentatively identified organic chemicals detected. All the groundwater managers believed the NTS results were useful and could help improve their testing recommendations to private well users. Even with qualitative limitations, NTS results can be an effective and valuable tool to inform the public and governance stakeholders in decisions around groundwater quality management.

Vulnerability of wells in unconfined and confined aquifers to modern contamination from flood events.

Groundwater is a primary potable water supply for coastal North Carolina (NC), but the increased intensity of extreme rainfall events and floods may exacerbate surface and subsurface processes that contribute anthropogenic chemicals to wells in the major confined aquifers of this region. We evaluated groundwater for organic chemicals of emerging concern (CEC) and the presence of tritium using flooded and not-flooded wells in the NC Department of Environmental Quality well monitoring network across the NC Coastal Plain. Flooded wells experienced standing water around the well casing at least once during the study period. Tritium concentrations, which indicate modern water presence (water recharged after 1953), were significantly greater in groundwater from flooded wells than not-flooded wells. In confined aquifers, modern water was detected at greater depths in flooded wells (206 m) than not-flooded wells (100 m). Suspect-screening high resolution mass spectrometry (HRMS) analysis of 150 groundwater samples yielded a total of 382 unique organic chemicals. Each groundwater sample contained, on average, 19 tentatively identified chemicals from the NIST 20 mass spectral database (M1) and 9 USEPA ToxCast chemicals. The number of tentatively-identified chemicals per sample was not significantly different among aquifers demonstrating the pervasive presence of the detected CECs in unconfined and confined aquifers. The presence of modern water in groundwater from flooded wells coincided with higher detection frequencies of certain organic contaminant classes, particularly pharmaceuticals, food additives, and regulated aromatic hydrocarbons. These results indicate that wells in both unconfined and confined aquifers are susceptible to modern water contamination during flood events; this finding has critical public health implications for coastal communities.

Locomotion and brain gene expression exhibit sex-specific non-monotonic dose-response to HFPO-DA during Drosophila melanogaster lifespan.

BACKGROUND: Legacy per- and polyfluoroalkyl substances (PFAS), known for their environmental persistence and bio-accumulative properties, have been phased out in the U.S. due to public health concerns. A newer polymerization aid used in the manufacture of some fluoropolymers, hexafluoropropylene oxide-dimer acid (HFPO-DA), has lower reported bioaccumulation and toxicity, but is a potential neurotoxicant implicated in dopaminergic neurodegeneration. OBJECTIVE: We investigated HFPO-DA's bio-accumulative potential and sex-specific effects on lifespan, locomotion, and brain gene expression in fruit flies. METHODS: We quantified bioaccumulation of HFPO-DA in fruit flies exposed to 8.7 × 104 µg/L of HFPO-DA in the fly media for 14 days via UHPLC-MS. Long-term effect on lifespan was determined by exposing both sexes to 8.7 × 102 - 8.7 × 105 µg/L of HFPO-DA in media. Locomotion was measured following 3, 7, and 14 days of exposures at 8.7 × 101 - 8.7 × 105 µg/L of HFPO-DA in media, and high-throughput 3'-end RNA-sequencing was used to quantify gene expression in fly brains across the same time points. RESULTS: Bioaccumulation of HFPO-DA in fruit flies was not detected. HFPO-DA-induced effects on lifespan, locomotion, and brain gene expression, and lowest adverse effect level (LOAEL) showed sexually dimorphic patterns. Locomotion scores significantly decreased in at least one dose at all time points for females and only at 3-day exposure for males, while brain gene expression exhibited non-monotonic dose-response. Differentially expressed genes correlated to locomotion scores revealed sex-specific numbers of positively and negatively correlated genes per functional category. CONCLUSION: Although HFPO-DA effects on locomotion and survival were significant at doses higher than the US EPA reference dose, the brain transcriptomic profiling reveals sex-specific changes and neurological molecular targets; gene enrichments highlight disproportionately affected categories, including immune response: female-specific co-upregulation suggests potential neuroinflammation. Consistent sex-specific exposure effects necessitate blocking for sex in experimental design during HFPO-DA risk assessment.

Toxicity assessment of hexafluoropropylene oxide-dimer acid on morphology, heart physiology, and gene expression during zebrafish (Danio rerio) development.

Hexafluoropropylene oxide-dimer acid (HFPO-DA) is one of the emerging replacements for the "forever" carcinogenic and toxic long-chain PFAS. HFPO-DA is a polymerization aid used for manufacturing fluoropolymers, whose global distribution and undetermined toxic properties are a concern regarding human and ecological health. To assess embryotoxic potential, zebrafish embryos were exposed to HFPO-DA at concentrations of 0.5-20,000 mg/L at 24-, 48-, and 72-h post-fertilization (hpf). Heart rate increased significantly in embryos exposed to 2 mg/L and 10 mg/L HFPO-DA across all time points. Spinal deformities and edema phenotypes were evident among embryos exposed to 1000-16,000 mg/L HFPO-DA at 72 hpf. A median lethal concentration (LC50) was derived as 7651 mg/L at 72 hpf. Shallow RNA sequencing analysis of 9465 transcripts identified 38 consistently differentially expressed genes at 0.5 mg/L, 1 mg/L, 2 mg/L, and 10 mg/L HFPO-DA exposures. Notably, seven downregulated genes were associated with visual response, and seven upregulated genes were expressed in or regulated the cardiovascular system. This study identifies biological targets and molecular pathways affected during animal development by an emerging, potentially problematic, and ubiquitous industrial chemical.

Non-target and suspect-screening analyses of hydroponic soybeans and passive samplers exposed to different watershed irrigation sources.

Water scarcity increases the likelihood of irrigating food crops with municipal wastewater that may pose potential dietary risks of regulated and non-regulated organic chemical uptake to edible plant tissues. Only a few studies have used high resolution mass spectrometry (HRMS) to assess the uptake of chemicals of concern into food crops. This study used non-target and suspect-screening analyses to compare total chemical features, tentatively identified chemicals (TICs), and EPA ToxCast chemicals in soybean plants and passive samplers exposed to five different irrigation sources that were collected from an agricultural watershed during mild drought conditions. Secondary-treated municipal wastewater effluent, two surface waters, two ground waters, and deionized municipal tap water were used for two hydroponic experiments: soybean roots and shoots and Composite Integrative Passive Samplers (CIPS) harvested after fourteen days of exposure and soybeans after fifty-six days. CIPS were sealed in separate glass amber jars to evaluate their efficacy to mimic chemical features, TICs, and ToxCast chemical uptake in plant roots, shoots, and beans. Total soybean biomass and water use were greatest for tap water, municipal wastewater, and surface water downstream of the municipal wastewater facility relative to groundwater samples and surface water collected upstream of the wastewater facility. ToxCast chemicals were ubiquitous across watershed irrigation sources in abundance, chemical use category, and number. Wastewater-exposed soybeans had the fewest extractable TICs in plant tissues of all irrigation sources. More ToxCast chemicals were identified in CIPS than extracted from irrigation sources by solid phase extraction. ToxCast chemicals in beans and CIPS were similar in number, chemical use category, and log Kow range. CIPS appear to serve as a useful surrogate for ToxCast chemical uptake in beans, the edible food product.

Cardiac physiology and metabolic gene expression during late organogenesis among F. heteroclitus embryo families from crosses between pollution-sensitive and -resistant parents.

BACKGROUND: The teleost fish Fundulus heteroclitus inhabit estuaries heavily polluted with persistent and bioaccumulative chemicals. While embryos of parents from polluted sites are remarkably resistant to toxic sediment and develop normally, embryos of parents from relatively clean estuaries, when treated with polluted sediment extracts, are developmentally delayed, displaying deformities characteristic of pollution-induced embryotoxicity. To gain insight into parental effects on sensitive and resistant phenotypes during late organogenesis, we established sensitive, resistant, and crossed embryo families using five female and five male parents from relatively clean and predominantly PAH-polluted estuaries each, measured heart rates, and quantified individual embryo expression of 179 metabolic genes. RESULTS: Pollution-induced embryotoxicity manifested as morphological deformities, significant developmental delays, and altered cardiac physiology was evident among sensitive embryos resulting from crosses between females and males from relatively clean estuaries. Significantly different heart rates among several geographically unrelated populations of sensitive, resistant, and crossed embryo families during late organogenesis and pre-hatching suggest site-specific adaptive cardiac physiology phenotypes relative to pollution exposure. Metabolic gene expression patterns (32 genes, 17.9%, at p < 0.05; 11 genes, 6.1%, at p < 0.01) among the embryo families indicate maternal pollutant deposition in the eggs and parental effects on gene expression and metabolic alterations. CONCLUSION: Heart rate differences among sensitive, resistant, and crossed embryos is a reliable phenotype for further explorations of adaptive mechanisms. While metabolic gene expression patterns among embryo families are suggestive of parental effects on several differentially expressed genes, a definitive adaptive signature and metabolic cost of resistant phenotypes is unclear and shows unexpected sensitive-resistant crossed embryo expression profiles. Our study highlights physiological and metabolic gene expression differences during a critical embryonic stage among pollution sensitive, resistant, and crossed embryo families, which may contribute to underlying resistance mechanisms observed in natural F. heteroclitus populations living in heavily contaminated estuaries.

Understanding the influence of multiple pollutant stressors on the decline of freshwater mussels in a biodiversity hotspot.

The Clinch River watershed of the upper Tennessee River Basin of Virginia and Tennessee, USA supports one of North America's greatest concentrations of freshwater biodiversity, including 46 extant species of native freshwater mussels (Order Unionida), 20 of which are protected as federally endangered. Despite the global biological significance of the Clinch River, mussel populations are declining in some reaches, both in species richness and abundance. The aim of this study was to evaluate the exposure of adult resident mussels to a suite of inorganic and organic contaminant stressors in distinct sections of the Clinch River that encompassed a range of mussel abundance and health. To provide insight into the potential role of pollutants in the decline of mussels, including within a previously documented "zone of mussel decline", the mainstem Clinch River (8 sites) and its tributaries (4 sites) were examined over two consecutive years. We quantified and related metals and organic contaminant concentrations in mussels to their associated habitat compartments (bed sediment, suspended particulate sediment, pore water, and surface water). We found that concentrations of organic contaminants in resident mussels, particularly the suite of 42 polycyclic aromatic hydrocarbons (PAHs) analyzed, were related to PAH concentrations in all four habitat (media) compartments. Further, PAH concentrations in mussel tissue (range 37.8-978.1 ng/g dry weight in 2012 and 194.3-1073.7 ng/g dry weight in 2013) were negatively related to the spatial pattern in mussel densities (rs = -0.64, p ≤ 0.05 in 2012 and rs = -0.83, p ≤ 0.05 in 2013) within the river, and were highest in the "zone of mussel decline". In contrast, the suite of 22 metals analyzed in resident mussels were largely unrelated to the spatial pattern of variation of metals in the four habitat compartments except for Manganese (Mn; range 3630.5-23,749.2 µg/g dry weight in 2012 and 1540.4-12,605.8 µg/g dry weight in 2013) in surface water (rs = 0.58, p < 0.1) and pore water (rs = 0.76, p ≤ 0.05). This study revealed that PAHs and Mn are important pollutant stressors to mussels in the Clinch River and that they are largely being delivered through the Guest River tributary watershed. Accordingly, future conservation and management efforts would benefit by identifying, and ideally mitigating, the sources of PAHs, Mn, and other current or legacy mining-associated pollutants to the mainstem river and its tributaries.

PAH-pollution effects on sensitive and resistant embryos: Integrating structure and function with gene expression.

Polycyclic aromatic hydrocarbons (PAHs) are among the most widespread natural and anthropogenic pollutants, and some PAHs are proven developmental toxicants. We chemically characterized clean and heavily polluted sites and exposed fish embryos to PAH polluted sediment extracts during four critical developmental stages. Embryos were collected from Fundulus heteroclitus populations inhabiting the clean and heavily polluted Superfund estuary. Embryos of parents from the clean sites are sensitive to PAH pollutants while those of parents from the heavily polluted site are resistant. Chemical analysis of embryos suggests PAH accumulation and pollution-induced toxicity among sensitive embryos during development that ultimately kills all sensitive embryos before hatching, while remarkably, the resistant embryos develop normally. The adverse effects on sensitive embryos are manifested as developmental delays, reduced heart rates, and severe heart, liver, and kidney morphological abnormalities. Gene expression analysis of early somitogenesis, heartbeat initiation, late organogenesis, and pre-hatching developmental stages reveals genes whose expression significantly differs between sensitive and resistant embryo populations and helps to explain mechanisms of sensitivity and resistance to polluted environments during vertebrate animal development.

Suspect-screening analysis of a coastal watershed before and after Hurricane Florence using high-resolution mass spectrometry.

On September 14, 2018, Hurricane Florence delivered ~686 mm rainfall to a 106 km2 watershed in coastal North Carolina, USA. A forested land treatment site comprises one third of the watershed wherein municipal wastewater effluent is spray-irrigated onto 8.9 km2 of forest. This communication provides insight for land treatment function under excess water duress as well as changes in organic chemical composition in on- and off-site waters before (June 2018) and after (September & December 2018) Hurricane Florence's landfall. We compare the numbers and relative abundances of chemical features detected using suspect screening high resolution mass spectrometry in waste-, ground-, and surface water samples. Values for upstream and receiving waters in September were lower than for sampling events in June and December, indicating an expected dilution effect across the watershed. Chemical diversity was greatest for all surface water samples in December, but only upstream surface water showed a dramatic five-fold increase in relative chemical abundance. Chemical abundance in on-site water and downstream surface water was equal to or lower than the September storm dilution effect. These data suggest that the land treatment system is functionally and hydrologically robust to extreme storm events and contributed to dilution of upstream chemical reservoirs for downstream receiving waters for months after the storm. Similar systems may embody one water reuse strategy robust to the increasing occurrence of extreme precipitation events.

Expanded coverage of non-targeted LC-HRMS using atmospheric pressure chemical ionization: a case study with ENTACT mixtures.

Non-targeted analysis (NTA) is a rapidly evolving analytical technique with numerous opportunities to improve and expand instrumental and data analysis methods. In this work, NTA was performed on eight synthetic mixtures containing 1264 unique chemical substances from the U.S. Environmental Protection Agency's Non-Targeted Analysis Collaborative Trial (ENTACT). These mixtures were analyzed by atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI) using both positive and negative polarities for a total of four modes. Out of the 1264 ENTACT chemical substances, 1116 were detected in at least one ionization mode, 185 chemicals were detected using all four ionization modes, whereas 148 were not detected. Forty-four chemicals were detected only by APCI, and 181 were detected only by ESI. Molecular descriptors and physicochemical properties were used to assess which ionization type was preferred for a given compound. One ToxPrint substructure (naphthalene group) was found to be enriched in compounds only detected using APCI, and eight ToxPrints (e.g., several alcohol moieties) were enriched in compounds only detected using ESI. Examination of physicochemical parameters for ENTACT chemicals suggests that those with higher aqueous solubility preferentially ionized by ESI-. While ESI typically detects a larger number of compounds, APCI offers chromatograms with less background, fewer co-elutions, and additional chemical space coverage, suggesting both should be considered for broader coverage in future NTA research. Graphical abstract.

Development of a computational method to quantify the partitioning of polycyclic aromatic hydrocarbons in seawater into dissolved and droplet forms.

The ecological risk and potential injury from polycyclic aromatic hydrocarbons (PAHs) from spilled crude oil in water is dependent on whether they are dissolved or associated with droplets/particles. Using oil collected from the Deepwater Horizon incident, laboratory experiments were conducted to develop a computational method to determine the physical state of PAHs in sampled seawater and compare these to results from methods designed to physically separate dissolved and droplet oil used during the spill. The analytical results were used to develop a method to calculate droplet/particulate concentrations from unfractionated water samples based on oil composition and allocation of minimally soluble oil components to the droplet fraction. The computational method is less labor-intensive and costly than field fractionation and can be used to optimize the use of previously collected data. The results also showed that physical separation can result in insoluble high-molecular-weight PAHs being present in the filtrate (i.e., the "dissolved" fraction), potentially leading to an overestimate of dissolved components.

Effect of silver nanoparticles on gill membranes of common carp: Modification of fatty acid profile, lipid peroxidation and membrane fluidity.

Although the toxicity of silver nanoparticles (AgNPs) in aquatic organisms has been extensively investigated, the mechanism by which AgNPs damage membranes remains unclear. This study investigated the toxic effects of a series of sub-lethal concentrations of AgNPs on the membranes of freshwater carp (Cyprinus carpio) gills, based on changes in membrane fatty acid (FA) profile, membrane fluidity, membrane lipid peroxidation, and histopathology. Most of the FAs in fish gill membrane was not significantly affected by exposure to multiple AgNPs concentrations, only few significant changes occurred in some specific FAs species at a high concentration of AgNPs exposure. In particular, high concentrations of AgNPs significantly decreased the proportions of two important long-chain n-3 series polyunsaturated FAs (C20: 5n3, and C22: 6n3), resulting in a decreased ratio of n-3 polyunsaturated FAs to n-6 polyunsaturated FAs (Σn-3UFA/Σn-6UFA). The AgNPs also caused a dose-dependent decrease in fish gill membrane fluidity, increased the level of lipid peroxidation, and inhibited Na+/K+-ATPase enzyme activity. Further histopathological examination revealed that exposure to AgNPs can cause toxic responses in the lamellae, including the thinning of the basement membrane, malformation, and inflammation. Together, the results suggest that the mechanism of AgNPs membrane toxicity involves the oxidization of long-chain omega-3 unsaturated FAs to saturated FAs via lipid peroxidation, resulting in, decreased membrane fluidity and ultimately the destruction of the normal physiological function of the fish gill membrane. The findings contribute significantly to our understanding of nanoparticle-induced membrane toxicity and potential risks in aquatic environments.

Suspect screening and prioritization of chemicals of concern (COCs) in a forest-water reuse system watershed.

Much research has assessed organic chemicals of concern (COCs) in municipal wastewater and receiving waters, but few studies have examined COCs in land treatment systems. Many prior studies have implemented targeted methods that quantify a relatively small fraction of COCs present in wastewater and receiving waters. This study used suspect screening to assess chemical features in ground- and surface waters from a watershed where secondary-treated wastewater is irrigated onto 900 ha of temperate forest, offering a more holistic view of chemicals that contribute to the exposome. Chemical features were prioritized by abundance and ToxPi scoring across seasonal sampling events to determine if the forest-water reuse system contributed to the chemical exposome of ground- and surface waters. The number of chemical features detected in wastewater was usually higher than on- and off-site ground- and surface waters; in wastewater, chemical features trended with precipitation in which greater numbers of features were detected in months with low precipitation. The number of chemical features detected in off- and on-site waters was similar. The lower overlap between chemical features found in wastewater and downstream surface waters, along with the similar numbers of features being detected in upstream and downstream surface waters, suggests that though wastewater may be a source of chemicals to ground and surface waters on-site, dissipation of wastewater-derived features (in number and peak area abundance) likely occurs with limited off-site surface water export by the forested land treatment system. Further, the numbers of features detected on site and the overlap between wastewater and surface waters did not increase during periods of low rainfall, counter to our initial expectations. The chemical features tentatively identified in this watershed appear common to features identified in other studies, warranting further examination on the potential for resulting impacts of these on humans and the environment.

Investigation of ciguatoxins in invasive lionfish from the greater caribbean region: Implications for fishery development.

Lionfish, native to reef ecosystems of the tropical and sub-tropical Indo-Pacific, were introduced to Florida waters in the 1980s, and have spread rapidly throughout the northwestern Atlantic, Caribbean Sea and the Gulf of Mexico. These invasive, carnivorous fish significantly reduce other fish and benthic invertebrate biomass, fish recruitment, and species richness in reef ecosystems. Fisheries resource managers have proposed the establishment of a commercial fishery to reduce lionfish populations and mitigate adverse effects on reef communities. The potential for a commercial fishery for lionfish is the primary reason to identify locations where lionfish accumulate sufficient amounts of ciguatoxin (CTX) to cause ciguatera fish poisoning (CFP), the leading cause of non-bacterial seafood poisoning associated with fish consumption. To address this issue, an initial geographic assessment of CTX toxicity in lionfish from the Caribbean and Gulf of Mexico was conducted. Lionfish samples (n = 293) were collected by spearfishing from 13 locations (74 sampling sites) around the Caribbean and Gulf of Mexico between 2012 and 2015. The highest frequencies of lionfish containing measurable CTX occurred in areas known to be high-risk regions for CFP in the central to eastern Caribbean (e.g., 53% British Virgin Islands and 5% Florida Keys). Though measurable CTX was found in some locations, the majority of the samples (99.3%) contained CTX concentrations below the United States Food and Drug Administration guidance level of 0.1 ppb Caribbean ciguatoxin-1 (C-CTX-1) equivalents (eq.). Only 0.7% of lionfish tested contained more than 0.1 ppb C-CTX-1 eq. As of 2018, there has been one suspected case of CFP from eating lionfish. Given this finding, current risk reduction techniques used to manage CTX accumulating fish are discussed.

Relation of contaminants to fish intersex in riverine sport fishes.

Endocrine active compounds (EACs) are pollutants that have been recognized as an emerging and widespread threat to aquatic ecosystems globally. Intersex, the presence of female germ cells within a predominantly male gonad, is considered a biomarker of endocrine disruption caused by EACs. We measured a suite of EACs and assessed their associated impacts on fish intersex occurrence and severity in a large, regulated river system in North Carolina and South Carolina, USA. Our specific objective was to determine the relationship of contaminants in water, sediment, and fish tissue with the occurrence and severity of the intersex condition in wild, adult black bass (Micropterus), sunfish (Lepomis), and catfish (Ictaluridae) species at 11 sites located on the Yadkin-Pee Dee River. Polycyclic aromatic hydrocarbons (PAHs), ethinylestradiol (EE2), and heavy metals were the most prevalent contaminants that exceeded effect levels for the protection of aquatic organisms. Fish intersex condition was most frequently observed and most severe in black basses and was less frequently detected and less severe in sunfishes and catfishes. The occurrence of the intersex condition in fish showed site-related effects, rather than increasing longitudinal trends from upstream to downstream. Mean black bass and catfish tissue contaminant concentrations were higher than that of sunfish, likely because of the latter's lower trophic position in the food web. Principal component analysis identified waterborne PAHs as the most correlated environmental contaminant with intersex occurrence and severity in black bass and sunfish. As indicated by the intersex condition, EACs have adverse but often variable effects on the health of wild sport fishes in this river, likely due to fluctuations in EAC inputs and the dynamic nature of the riverine system. These findings enhance the understanding of the relationship between contaminants and fish health and provide information to guide ecologically comprehensive conservation and management decisions.

Cadmium disrupts signaling of the hypoxia-inducible (HIF) and transforming growth factor (TGF-ß) pathways in placental JEG-3 trophoblast cells via reactive oxygen species.

Epidemiologic studies indicate an association between exposure to cadmium (Cd) and placental-related pregnancy disorders. While a precise mechanism is unknown, oxidative imbalance and dysregulation of the hypoxia inducible factor (HIF) and transforming growth factor beta (TGF-ß) pathways have been implicated in placental disease pathogenesis. Here we investigated key oxidative and placentation pathways in JEG-3 placental trophoblast cells treated with Cd alone, environmental water samples predominated by Cd with low concentrations of other metals (e.g. inorganic arsenic (iAs)) collected from a waste-site, and a matched mixture of Cd and iAs prepared in the laboratory. The induction of cytosolic reactive oxygen species (ROS), expression of metallothionein (MT) isoforms, HIF1α and downstream targets, and expression of TGFß pathway-associated genes and proteins were assessed. Additionally, the effect of pre-treatment with the antioxidant N-acetyl cysteine (NAC) on ROS generation and effects on HIF, MT and TGF-ß signaling pathways was examined. Cd and Cd-mixture treated cells displayed higher levels of ROSs with accompanying disruption of HIF and TGFß pathway signaling versus controls, with the Cd-mixture eliciting a greater effect. Conversely, pretreatment with NAC reduced Cd-induced ROS production and disruption of HIF, MT and TGFß pathway signaling. The results indicate that treatment of placental trophoblast cells with Cd results in increased production of ROSs that disrupt placentation pathways involved in disease pathogenesis. Also, co-occurrence of Cd with other toxic metals, particularly arsenic, may induce detrimental health effects that are currently underestimated when analyzed as single metals.

Comparison of emerging contaminants in receiving waters downstream of a conventional wastewater treatment plant and a forest-water reuse system.

Forest-water reuse (FWR) systems treat municipal, industrial, and agricultural wastewaters via land application to forest soils. Previous studies have shown that both large-scale conventional wastewater treatment plants (WWTPs) and FWR systems do not completely remove many contaminants of emerging concern (CECs) before release of treated wastewater. To better characterize CECs and potential for increased implementation of FWR systems, FWR systems need to be directly compared to conventional WWTPs. In this study, both a quantitative, targeted analysis and a nontargeted analysis were utilized to better understand how CECs release to waterways from an FWR system compared to a conventional treatment system. Quantitatively, greater concentrations and total mass load of CECs was exhibited downstream of the conventional WWTP compared to the FWR. Average summed concentrations of 33 targeted CECs downstream of the conventional system were ~ 1000 ng/L and downstream of the FWR were ~ 30 ng/L. From a nontargeted chemical standpoint, more tentatively identified chemicals were present, and at a greater relative abundance, downstream of the conventional system as well. Frequently occurring contaminants included phthalates, pharmaceuticals, and industrial chemicals. These data indicate that FWR systems represent a sustainable wastewater treatment alternative and that emerging contaminant release to waterways was lower at a FWR system than a conventional WWTP.

Contaminants in tropical island streams and their biota.

Environmental contamination is problematic for tropical islands due to their typically dense human populations and competing land and water uses. The Caribbean island of Puerto Rico (USA) has a long history of anthropogenic chemical use, and its human population density is among the highest globally, providing a model environment to study contaminant impacts on tropical island stream ecosystems. Polycyclic Aromatic Hydrocarbons, historic-use chlorinated pesticides, current-use pesticides, Polychlorinated Biphenyls (PCBs), and metals (mercury, cadmium, copper, lead, nickel, zinc, and selenium) were quantified in the habitat and biota of Puerto Rico streams and assessed in relation to land-use patterns and toxicological thresholds. Water, sediment, and native fish and shrimp species were sampled in 13 rivers spanning broad watershed land-use characteristics during 2009-2010. Contrary to expectations, freshwater stream ecosystems in Puerto Rico were not severely polluted, likely due to frequent flushing flows and reduced deposition associated with recurring flood events. Notable exceptions of contamination were nickel in sediment within three agricultural watersheds (range 123-336ppm dry weight) and organic contaminants (PCBs, organochlorine pesticides) and mercury in urban landscapes. At an urban site, PCBs in several fish species (Mountain Mullet Agonostomus monticola [range 0.019-0.030ppm wet weight] and American Eel Anguilla rostrata [0.019-0.031ppm wet weight]) may pose human health hazards, with concentrations exceeding the U.S. Environmental Protection Agency (EPA) consumption limit for 1 meal/month. American Eel at the urban site also contained dieldrin (range < detection-0.024ppm wet weight) that exceeded the EPA maximum allowable consumption limit. The Bigmouth Sleeper Gobiomorous dormitor, an important piscivorus sport fish, accumulated low levels of organic contaminants in edible muscle tissue (due to its low lipid content) and may be most suitable for human consumption island-wide; only mercury at one site (an urban location) exceeded EPA's consumption limit of 3 meals/month for this species. These results comprise the first comprehensive island-wide contaminant assessment of Puerto Rico streams and biota and provide natural resource and public health agencies here and in similar tropical islands elsewhere with information needed to guide ecosystem and fisheries conservation and management and human health risk assessment.

A Retrospective Analysis of Agricultural Herbicides in Surface Water Reveals Risk Plausibility for Declines in Submerged Aquatic Vegetation.

The Albemarle-Pamlico Estuarine System (APES) is the second largest estuarine system within the mainland of the United States and is estimated to have lost about half of its submerged aquatic vegetation (SAV) over the past several decades. The issue of herbicide runoff and subsequent toxic effects to SAV is important because of the extensive agricultural production that occurs in the APES region. The aim of this study was to conduct a retrospective analysis of herbicide influx to waters of the APES region during the time period of documented SAV declines and to compare the measured concentrations to SAV toxicity thresholds and changes in agricultural land use. Surface water grab samples were collected at 26 sites in the APES region during May through July 2000. The most consistently measured herbicides were alachlor, atrazine, and metolachlor with geometric mean concentrations ranging from 29 to 2463 ng/L for alachlor, 14 to 7171 ng/L for atrazine, and 17 to 5866 ng/L for metolachlor. Concentrations of alachlor, atrazine, and metolachlor measured in water samples from the APES region in 2000 exceeded several of the established benchmarks, standards, or guidelines for protection of aquatic plants. Although this evaluation was of point-in-time herbicide samples (year 2000) and not analyzed for all possible herbicides used at the time, they were taken during the period of SAV declines, reveal the plausibility of exposure risk to SAV, and suggest that herbicide runoff should be studied along with other variables that influence SAV growth and distribution in future studies.

Pharmaceuticals in a temperate forest-water reuse system.

Forest-water reuse systems infiltrate municipal, industrial, and agricultural wastewaters through forest soils to shallow aquifers that ultimately discharge to surface waters. Their ability to mitigate regulated nutrients, metals, and organic chemicals is well known, but the fate of non-regulated chemicals in these systems is largely unstudied. This study quantified 33 pharmaceuticals and personal care products (PPCPs) in soils, groundwaters, and surface waters in a 2000-hectare forest that receives ~1200mm/year of secondary-treated, municipal wastewater in addition to natural rainfall (~1300mm/year). This forest-water reuse system does contribute PPCPs to soils, groundwater, and surface waters. PPCPs were more abundant in soils versus underlying groundwater by an order of magnitude (5-10ng/g summed PPCPs in soil and 50-100ng/L in groundwater) and the more hydrophobic chemicals were predominant in soil over water. PPCP concentrations in surface waters were greater at the onset of significant storm events and during low-rainfall periods when total summed PPCPs were >80ng/L, higher than the annual average. With few exceptions, the margins of exposure for PPCPs in groundwater and surface waters were several orders of magnitude above values indicative of human health risk.