Laboratory-Derived Bioaccumulation Kinetic Parameters for Four Per- and Polyfluoroalkyl Substances in Freshwater Mussels.

Although freshwater mussels are imperiled and identified as key conservation priorities, limited bioaccumulation information is available on these organisms for contaminants of emerging concern. In the present study we investigated the bioaccumulation of per- and polyfluoroalkyl substances (PFAS) in the model freshwater pond mussel Sagittunio subrostratus because mussels provide important ecosystem services and are important components of aquatic systems where PFAS occur. In the present study we selected four representative perfluorinated carboxylic acids and sulfonic acids, then determined the bioaccumulation kinetics of freshwater mussels in a controlled laboratory study. Because uptake (ku ) and elimination (ke ) rate constants and time to steady state are important parameters for food web bioaccumulation models, we derived bioaccumulation kinetic parameters following exposure to perfluorohexane sulfonic acid (PFHxS), perfluorooctane sulfonic acid (PFOS), and perfluorodecanoic acid (PFDA) at 10 µg/L and perfluoroundecanoic acid (PFUnDA) at 1 µg/L during a 14-day uptake period followed by a 7-day elimination period. Kinetic and ratio-based bioaccumulation factors (BAFs) were subsequently calculated, for example ratio-based BAFs for mussel at day 7 were determined for PFHxS (0.24 ± 0.08 L/kg), PFOS (7.73 ± 1.23 L/kg), PFDA (4.80 ± 1.21 L/kg), and PFUnDA (84.0 ± 14.4 L/kg). We generally observed that, for these four model PFAS, freshwater mussels have relatively low BAF values compared with other aquatic invertebrates and fish. Environ Toxicol Chem 2023;42:1190-1198. © 2023 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Bioaccumulation Kinetics of Model Pharmaceuticals in the Freshwater Unionid Pondmussel, Sagittunio subrostratus.

Bioaccumulation of ionizable pharmaceuticals has been increasingly studied, with most reported aquatic tissue concentrations in field or laboratory experiments being from fish. However, higher levels of antidepressants have been observed in bivalves compared with fish from effluent-dominated and dependent surface waters. Such observations may be important for biodiversity because approximately 70% of freshwater bivalves in North America are considered to be vulnerable to extinction. Because experimental bioaccumulation information for freshwater bivalves is lacking, we examined accumulation dynamics in the freshwater pondmussel, Sagittunio subrostratus, following exposure to a model weak acid, acetaminophen (mean (±SD) = 4.9 ± 1 µg L-1 ), and a model weak base, sertraline (mean (±SD) = 1.1 ± 1.1 µg L-1 ) during 14-day uptake and 7-day depuration experiments. Pharmaceutical concentrations were analyzed in water and tissue using isotope dilution liquid chromatography-tandem mass spectrometry. Mussels accumulated two orders of magnitude higher concentrations of sertraline (31.7 ± 9.4 µg g-1 ) compared to acetaminophen (0.3 ± 0.1 µg g-1 ). Ratio and kinetic-based bioaccumulation factors of 28,836.4 (L kg-1 ) and 34.9 (L kg-1 ) were calculated for sertraline and for acetaminophen at 65.3 (L kg-1 ) and 0.13 (L kg-1 ), respectively. However, after 14 days sertraline did not reach steady-state concentrations, although it was readily eliminated by S. subrostratus. Acetaminophen rapidly reached steady-state conditions but was not depurated over a 7-day period. Future bioaccumulation studies of ionizable pharmaceuticals in freshwater bivalves appear warranted. Environ Toxicol Chem 2023;42:1183-1189. © 2023 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Development of a standardized adsorbable organofluorine screening method for wastewaters with detection by combustion ion chromatography.

Per- and polyfluoroalkyl substances (PFAS) are man-made organofluorine chemicals that can contaminate environmental waters and have gained worldwide attention over the past two decades. PFAS are most frequently detected by mass spectrometric targeted analysis methods which may not detect all the PFAS in samples. This report describes the investigation of adsorbable organofluorine (AOF) with detection by combustion ion chromatography (CIC) for detection of PFAS in surface waters and wastewaters that adsorb to granular activated carbon (GAC) with the recognition that this technique measures more than just PFAS. Overall mean recoveries of 77-120% were obtained in 17 of the 18 tested surface water and wastewater matrices spiked with perfluoropentane sulfonate (PFPeS) and 55-119% mean recoveries were obtained in 11 of the 12 surface water and wastewater matrices spiked with a PFAS mixture. Poor method performance (34-39% mean recoveries) was observed in landfill leachate wastewater. Method detection limits of 1.4-2.2 µg L-1 were achieved using 100 mL sample volumes adsorbed onto commercially available GAC. This report demonstrates that this AOF technique can be a useful screening tool for estimating organofluorine concentrations when PFAS contamination is suspected.

Interaction of the Olfactory System of Rainbow Trout (Oncorhynchus mykiss) with Diltiazem.

Diltiazem is ubiquitously prescribed and has been reported in many effluents and freshwater bodies. Being a calcium channel blocker, diltiazem could disrupt the function of the sensory and central nervous systems. In the present study, using electro-olfactography (EOG), we investigated the interaction of diltiazem with the olfactory sensory neurons (OSNs) of rainbow trout by looking into the detection threshold and effects of immediate (~5 min) and acute (24 h) exposure to diltiazem at 6.6, 66, and 660 µg/L. We also studied the accumulation of the drug in fish plasma and whole body. Brief exposure to diltiazem impaired the OSN response to a chemosensory stimulus in a concentration-dependent manner at 6.6 µg/L and higher, whereas OSNs exposed for 24 h only displayed an impairment at 660 µg/L. Chemical analysis showed that the accumulation of diltiazem in fish plasma and body correlated with the EOG response because it was 10 times higher in the group that displayed a significant impairment (660 µg/L) compared to the other 2 groups (6.6, 66 µg/L). This correlation suggests that the impact of diltiazem on OSNs might partially be through the accumulated molecules in the fish bloodstream. Fish did not detect diltiazem as a sensory stimulus even at concentrations as high as 660 µg/L; thus, fish could potentially swim toward or fail to escape harmful concentrations of diltiazem. Environ Toxicol Chem 2022;41:554-550. © 2020 SETAC.

Plasma Vitellogenin Reveals Potential Seasonal Estrogenicity in Fish from On-Site Wastewater Treatment Systems in Semi-Arid Streams Influenced by Snowmelt.

Effluents from on-site wastewater treatment systems can influence surface water quality, particularly when infrastructure is aging, malfunctioning, and improperly installed. Municipal wastewater often contains chemical compounds that can lead to adverse biological effects, such as reproductive impairment, in organisms that are chronically exposed. A significant number of these compounds are endocrine-disrupting chemicals. Water quality influences of on-site systems are poorly studied in semi-arid regions where instream flows are seasonally dependent on snowmelt, and when instream dilution of wastewater effluents is minimal during other times of the year. Here we examined surface water estrogenicity in low order tributaries of two unique semi-arid streams with on-site wastewater treatment systems, for which seasonal instream flow fluctuations occur in Park City, UT, USA. Water samples were collected from a total of five locations along two lotic systems downstream from active on-site treatment systems. Samples were extracted for targeted chemical analyses and to perform in vivo and in vitro bioassays with juvenile rainbow trout. Estrogenic activity was measured by quantifying the concentration and expression of vitellogenin (VTG) in plasma and liver, respectively. Plasma VTG presented elevated levels in fish exposed to water samples collected at the two sites in close proximity to on-site systems and during seasons with low stream discharge, though the levels observed did not suggest severe endocrine disruption. However, long-term exposure to these surface water could compromise the fish populations. While the sensitivity of in vitro bioassays was low and targeted chemical analyses did not identify causative compounds, the use of complementary lines of evidence (e.g., in vivo biological models) was advantageous in identifying estrogenic activity in waters influenced by effluents from on-site wastewater systems.

Multi-approach assessment for the evaluation of spatio-temporal estrogenicity in fish from effluent-dominated surface waters under low instream flow.

Current practices employed by most wastewater treatment plants (WWTP) are unable to completely remove endocrine disrupting compounds (EDCs) from reclaimed waters, and consistently discharge these substances to receiving systems. Effluent-dominated and dependent surface waters, especially during low instream flows, can increase exposure and risks to aquatic organisms due to adverse biological effects associated with EDCs. Given the ecological implications that may arise from exposure to such compounds, the present a multi-approach study examined spatio-temporal estrogenic potential of wastewater effluent to fish in East Canyon Creek (ECC), Utah, USA, a unique urban river with instream flows seasonally influenced by snowmelt. Juvenile rainbow trout (Oncorhynchus mykiss) were caged at different upstream and downstream sites from an effluent discharge during the summer and fall seasons. In the summer, where approximately 50% of the streamflow was dominated by effluent, fish from the upstream and a downstream site, located 13 miles away from the effluent discharge, presented significantly elevated concentrations of plasma vitellogenin (VTG). Similarly, significantly high 17ß-estradiol to 11-ketotestosterone ratios were measured in the summer across all sites and time points, compared to the fall. In the laboratory, juvenile fish and primary hepatocytes were exposed to concentrated effluent and surface water samples. Quantification of VTG, although in significantly lower levels, resembled response patterns observed in fish from the field study. Furthermore, analytical quantification of common EDCs in wastewater revealed the presence of estriol and estrone, though these did not appear to be related to the observed biological responses, as these were more significant in sites were no EDCs were detected. These combined observations suggest potential estrogenicity for fish in ECC under continuous exposures and highlight the advantages of following weight-of-evidence (WoE) approaches for environmental monitoring, as targeted analytically-based assessments may or may not support the identification of causative contaminants for adverse biological effects.

Spatial and seasonal occurrence of semi-volatile organic compounds (SVOCs) in fish influenced by snowmelt and municipal effluent discharge.

In the present study we examined spatial and seasonal trends in the levels of a wide suite of semi-volatile organic compounds (SVOCs) in brown trout (Salmo trutta) and mottled sculpin (Cottus bairdii) in East Canyon Creek, Utah, USA, an effluent-dominated stream during summer months. Fish samples were collected from four sampling sites, including one reference site upstream, and three sites at incremental distances downstream of the effluent discharge over multiple seasons. The samples were analyzed for 218 lipophilic contaminants, including pesticides and their metabolites, polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and polybrominated diphenyl ethers (PBDEs) and other flame retardants. Some PAHs, pesticides and their metabolites, PCBs, PBDEs and other flame retardants were measured in mottled sculpin (11 analytes) and brown trout (17 analytes). Hexachlorobenzene (HCB), p,p'-DDE, BDE-47 and triphenyl phosphate (TPHP) were the most frequently detected contaminants in mottled sculpin and brown trout, while BDE-47 and p,p'-DDE were measured at the highest concentrations, reaching up to 73 and 19 ng/g wet weight, respectively. Our results indicated that snowmelt did not alter accumulation of the examined lipophilic contaminants, and no consistent seasonal differences were observed in their accumulation. A spatial pattern was observed for PBDE congeners, where lowest levels were measured in fish tissues from a reference site, and highest concentrations were measured in fish collected downstream of the effluent discharge, indicating that municipal effluent discharge contributes to the elevated PBDE levels in fish residing in this effluent-dominated stream. We further calculated screening level consumption risks following United States Environmental Protection Agency (EPA) methods, and identified the importance of considering discharge gradients in effluent-dominated systems during bioaccumulation assessments.

Periphyton, bivalves and fish differentially accumulate select pharmaceuticals in effluent-dependent stream mesocosms.

Pharmaceuticals and other ionizable contaminants from municipal wastewater treatment plant effluent can bioaccumulate in fish, particularly in effluent dominated and dependent systems in semi-arid and arid regions. However, invertebrate bioaccumulation of these compounds has been less studied. Using municipal wastewater effluent as source water in outdoor stream mesocosms to simulate effluent-dependent lotic systems, we examined bioaccumulation of several widely-used pharmaceuticals including acetaminophen (nonsteroidal anti-inflamatory), caffeine (stimulant), carbamazepine (anti-epileptic), diltiazem (calcium channel blocker), diphenhydramine (anti-histamine), fluoxetine (anti-depressant), norfluoxetine (anti-depressant metabolite), and sertraline (anti-depressant) in freshwater clams (Corbicula fluminea), periphyton and stoneroller minnows (Campostoma anomalum), a commonly studied grazer in stream ecology, during a replicated outdoor stream mesocosm study at the Baylor Experimental Aquatic Research facility. Target analytes were determined in tissues, source effluent and stream water by isotope dilution LC-MS/MS. After an 8-day uptake period, clams accumulated a number of pharmaceuticals, including acetaminophen, carbamazepine, diltiazem, diphenhydramine, fluoxetine, norfluoxetine and sertraline with maximum concentrations reaching low µg/kg. We observed uptake rates in clams for acetaminophen at 2.8 µg/kg per day, followed by diphenhydramine (1.2 µg/kg per day) and carbamazepine (1.1 µg/kg per day). Caffeine, carbamazepine, diltiazem and diphenhydramine were measured in periphyton. Diphenhydramine was the only compound detected in all matrices, where bioaccumulation factors (BAFs) were elevated in bivalves (1631 ± 589 L/kg), compared to stoneroller minnows (247 ± 84 L/kg) and periphyton (315 ± 116 L/kg). Such BAF variability across multiple biological matrices highlight the need to understand bioaccumulation differences for ionizable contaminants among freshwater biota, including threatened and endangered species (e.g., unionids), commercially important bivalves (e.g., estuarine and marine bivalves), and fish.

Pharmaceutical uptake kinetics in rainbow trout: In situ bioaccumulation in an effluent-dominated river influenced by snowmelt.

Whether seasonal instream flow dynamics influence bioaccumulation of pharmaceuticals by fish is not well understood, specifically for urban lotic systems in semi-arid regions when flows are influenced by snowmelt. We examined uptake of select pharmaceuticals in rainbow trout (Oncorhynchus mykiss) caged in situ upstream and at incremental distances downstream (0.1, 1.4, 13 miles) from a municipal effluent discharge to East Canyon Creek in Park City, Utah, USA during summer and fall of 2018. Fish were sampled over 7-d to examine if uptake occurred, and to define uptake kinetics. Water and fish tissues were analyzed via isotope dilution liquid chromatography tandem mass spectrometry. Several pharmaceuticals were consistently detected in water, fish tissue and plasma, including carbamazepine, diphenhydramine, diltiazem, and fluoxetine. Pharmaceutical levels in water ranged up to 151 ng/L for carbamazepine, whereas the effluent tracer sucralose was consistently observed at low µg/L levels. During both summer and fall experiments at each of three downstream locations from effluent discharge, rainbow trout rapidly accumulated these pharmaceuticals; tissue levels reached steady state conditions within 24-96 h. Spatial and temporal differences for pharmaceutical levels in rainbow trout directly corresponded with surface water exposure concentrations, and uptake kinetics for individual pharmaceuticals did not vary among sites or seasons. Such observations are consistent with recent laboratory bioconcentration studies, which collectively indicate inhalational exposure from water governs rapid accumulation of ionizable base pharmaceuticals by fish in inland surface waters.

Toward Sustainable Environmental Quality: Priority Research Questions for Asia.

Environmental and human health challenges are pronounced in Asia, an exceptionally diverse and complex region where influences of global megatrends are extensive and numerous stresses to environmental quality exist. Identifying priorities necessary to engage grand challenges can be facilitated through horizon scanning exercises, and to this end we identified and examined 23 priority research questions needed to advance toward more sustainable environmental quality in Asia, as part of the Global Horizon Scanning Project. Advances in environmental toxicology, environmental chemistry, biological monitoring, and risk-assessment methodologies are necessary to address the adverse impacts of environmental stressors on ecosystem services and biodiversity, with Asia being home to numerous biodiversity hotspots. Intersections of the food-energy-water nexus are profound in Asia; innovative and aggressive technologies are necessary to provide clean water, ensure food safety, and stimulate energy efficiency, while improving ecological integrity and addressing legacy and emerging threats to public health and the environment, particularly with increased aquaculture production. Asia is the largest chemical-producing continent globally. Accordingly, sustainable and green chemistry and engineering present decided opportunities to stimulate innovation and realize a number of the United Nations Sustainable Development Goals. Engaging the priority research questions identified herein will require transdisciplinary coordination through existing and nontraditional partnerships within and among countries and sectors. Answering these questions will not be easy but is necessary to achieve more sustainable environmental quality in Asia. Environ Toxicol Chem 2020;39:1485-1505. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Antidepressants in Surface Waters: Fluoxetine Influences Mosquitofish Anxiety-Related Behavior at Environmentally Relevant Levels.

Pharmaceutical contamination is an increasing problem globally. In this regard, the selective serotonin reuptake inhibitors (SSRIs)-a group of antidepressants-are particularly concerning. By disrupting the serotonergic system, SSRIs have the potential to affect ecologically important behaviors in exposed wildlife. Despite this, the nature and magnitude of behavioral perturbations resulting from environmentally relevant SSRI exposure among species is poorly understood. Accordingly, we investigated the effects of two field-realistic levels of the SSRI fluoxetine (61 and 352 ng/L) on sociability and anxiety-related behaviors in eastern mosquitofish ( Gambusia holbrooki) for 28 days. Additionally, we measured whole-body tissue concentrations of fluoxetine and norfluoxetine. We found that fluoxetine altered anxiety-related behavior but not sociability. Specifically, female fish showed reduced anxiety-related behavior at the lower treatment level, while males showed an increase at the higher treatment level. In addition, we report a biomass-dependent and sex-specific accumulation of fluoxetine and norfluoxetine, with smaller fish showing higher relative tissue concentrations, with this relationship being more pronounced in males. Our study provides evidence for nonmonotonic and sex-specific effects of fluoxetine exposure at field-realistic concentrations. More broadly, our study demonstrated that neuroactive pharmaceuticals, such as fluoxetine, can affect aquatic life by causing subtle but important shifts in ecologically relevant behaviors.

Corbicula fluminea rapidly accumulate pharmaceuticals from an effluent dependent urban stream.

Freshwater bivalve populations are stressed by watershed development at the global scale. Though pharmaceuticals released from wastewater treatment plant effluent discharges are increasingly reported to bioaccumulate in fish, an understanding of bioaccumulation in bivalves is less defined. In the present study, we examined accumulation of 12 target pharmaceuticals in C. fluminea during a 42 day in situ study in Pecan Creek, an effluent dependent wadeable stream in north central Texas, USA. Caged clams were placed at increasing distances (5 m, 643 m, 1762 m) downstream from a municipal effluent discharge and then subsampled on study days 7, 14, 28 and 42. Acetaminophen, caffeine, carbamazepine, diltiazem, diphenhydramine, fluoxetine, norfluoxetine, sertraline, desmethylsertraline, and methylphenidate were identified in C. fluminea whole body tissue homogenates via isotope dilution liquid chromatography-tandem mass spectrometry. Tissue concentrations ranged from low µg/kg (methylphenidate) to 341 µg/kg (sertraline). By study day 7, rapid and apparent pseudo-steady state accumulation of study compounds was observed in clams; this observation continued throughout the 42 d study. Notably, elevated bioaccumulation factors (L/kg) for sertraline were observed between 3361 and 6845, which highlights the importance of developing predictive bioaccumulation models for ionizable contaminants with bivalves. Future research is also necessary to understand different routes of exposure and elimination kinetics for pharmaceutical accumulation in bivalves.

Spatial and temporal influence of onsite wastewater treatment systems, centralized effluent discharge, and tides on aquatic hazards of nutrients, indicator bacteria, and pharmaceuticals in a coastal bayou.

In the rapidly urbanizing watersheds and estuaries flowing to the Gulf of Mexico in Texas, USA, instream flows are increasingly influenced by point source and nonpoint source discharges. Spatial and temporal tidal influences on water quality, especially for contaminants of emerging concern (CECs), is poorly understood in estuaries and coastal systems. We selected Dickinson Bayou, an urban estuary in Galveston County, Texas, for study because it has historically impaired water quality, receives point source discharge from one major wastewater treatment plant (WWTP), while also being influenced by high densities of onsite sewage facilities upstream in the watershed. We explored the occurrence and potential hazards of aquatic contaminants, including nutrients, indicator bacteria for pathogens, and CECs, in relation to this point source discharge, across seasons and at high and low tides. Aquatic contaminants and associated hazards varied significantly in relation to the WWTP discharge, and were influenced by onsite systems. In fact, spatiotemporal water quality varied by class of contaminants (e.g., nutrients, indicator bacteria, CECs), which indicates that traditional surface water monitoring activities should account for such environmental complexity. This study provides a diagnostic approach for future studies of emerging water quality challenges across gradients of rapidly urbanizing coastal bays and estuaries.

Sex may influence environmental diphenhydramine accumulation in Round Stingrays.

Despite the amount of treated wastewater discharged into the Southern California Bight, few studies have examined pharmaceutical compounds in local biota. The Round Stingray (Urobatis halleri) was selected as a representative elasmobranch species to perform an exploratory study on environmental pharmaceutical exposure. Archived liver samples of males and females from juvenile to adult size classes from several locations (n = 53) were examined for 18 pharmaceutical and illicit drug compounds using isotope-dilution LC-MS/MS. Very few compounds were detected in stingray livers, with diphenhydramine as the only pharmaceutical above quantitation limits. Only stingrays collected from the urban site (mainland California) had detectable levels of diphenhydramine compared to no detections in reference stingrays (offshore island). Sex and sampling location substantially influenced both detection rate and concentrations. Our results suggest that aspects of species' ecology and physiology should be considered for future studies investigating pharmaceutical exposure in elasmobranchs.

Spatio-temporal bioaccumulation and trophic transfer of ionizable pharmaceuticals in a semi-arid urban river influenced by snowmelt.

Bioaccumulation of pharmaceuticals in aquatic organisms is increasingly reported in the peer-reviewed literature. However, seasonal instream dynamics including occurrence and bioaccumulation across trophic positions are rarely studied, particularly in semiarid streams with flows influenced by seasonal snowmelt and municipal effluent discharges. Thus, we selected East Canyon Creek in Park City, Utah, USA to examine spatio-temporal bioaccumulation of select ionizable pharmaceuticals across trophic positions using trophic magnification factors calculated at incremental distances (0.15, 1.4, 13 miles) downstream from a municipal effluent discharge during spring (May), Summer (August), and fall (October). Nine target analytes were detected in all species during all sampling events. Trophic dilution was consistently observed for amitriptyline, caffeine, diphenhydramine, diltiazem, fluoxetine, and sertraline, regardless of seasonal instream flows or distance from effluent discharge. Calculated TMFs ranged from 0.01-0.71 with negative slopes observed for all regressions of chemical residue in tissue and trophic position. We further presents the first empirical investigation of normalizing pharmaceutical concentrations to lipid, phospholipid or protein fractions using pair matched fish samples. Empirical results identify that normalization of ionizable pharmaceutical residues in aquatic tissues to neutral lipids, polar lipids, or the total protein fraction is inappropriate, though bioaccumulation studies examining influences of internal partitioning (e.g., plasma proteins) are needed.

At the Intersection of Urbanization, Water, and Food Security: Determination of Select Contaminants of Emerging Concern in Mussels and Oysters from Hong Kong.

Aquaculture, which is growing 3-5 times faster than terrestrial agriculture, will play an important role to meet future global food production needs. However, over 80% of global sewage production is returned to the environment untreated or poorly treated. In developing nations, these nontraditional waters of diverse quality are being recycled for aquaculture, yet chemical residues are differentially studied. Here, we examined pharmaceuticals, pesticides, polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), and flame retardants in marine bivalves using isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) and low-pressure gas chromatography-tandem mass spectrometry (LP GC-MS/MS). Green-lipped mussels from the field and oysters from aquaculture net pens, which are harvested as food products, were collected adjacent to point source municipal wastewater and landfill leachate effluent discharges, respectively, in Hong Kong, the fourth most densely populated country in the world. Multiple classes of pharmaceutical, pesticides, PAHs, and phosphorus-based flame retardants were detected at low µg/kg levels. Acceptable servings per week indicated minimal risk for a number of chemicals; however, such calculations could not be performed for other contaminants of emerging concern. Future efforts are needed to better understand contaminant influences on marine bivalve populations and aquaculture product safety, particularly in rapidly urbanizing regions of developing countries with limited wastewater infrastructure.

Acute exposure to an environmentally relevant concentration of diclofenac elicits oxidative stress in the culturally important galaxiid fish Galaxias maculatus.

Diclofenac is a nonsteroidal anti-inflammatory drug (NSAID) of growing concern in aquatic environments worldwide; nevertheless, knowledge of its effects on aquatic biota is restricted to a few model species with limited information regarding its mechanisms of impact. In the present study, diclofenac accumulation, its effects on metabolic rate, ionoregulation, and oxidative stress were examined at environmentally relevant (0.17 µg L-1 ) and elevated (763 µg L-1 ) concentrations in a culturally and economically important galaxiid fish, inanga (Galaxias maculatus), from the Southern Hemisphere. This species is among the most widespread freshwater fish in the world but its sensitivity to emerging contaminants is unknown. Following an acute 96-h exposure, bioconcentration of diclofenac was measured in the inanga whole-body, resulting in an estimated bioconcentration factor of 87 for the 0.17-µg L-1 exposure concentration, approaching values where transfer through the food chain should be considered. Lipid peroxidation in the liver was significantly elevated at both 0.17- and 763-µg L-1 exposure concentrations but lipid peroxidation in the kidney and gill decreased after diclofenac exposure. Catalase activity was also elevated in the liver of inanga but activity decreased in the gill. There were no effects of diclofenac on metabolic rate or ion (sodium and calcium) influx rates. These data indicate that toxicologically relevant adverse outcomes and bioconcentration of diclofenac at environmentally relevant levels warrant additional study in this important fish. Environ Toxicol Chem 2018;37:224-235. © 2017 SETAC.

Pharmaceuticals in water, fish and osprey nestlings in Delaware River and Bay.

Exposure of wildlife to Active Pharmaceutical Ingredients (APIs) is likely to occur but studies of risk are limited. One exposure pathway that has received attention is trophic transfer of APIs in a water-fish-osprey food chain. Samples of water, fish plasma and osprey plasma were collected from Delaware River and Bay, and analyzed for 21 APIs. Only 2 of 21 analytes exceeded method detection limits in osprey plasma (acetaminophen and diclofenac) with plasma levels typically 2-3 orders of magnitude below human therapeutic concentrations (HTC). We built upon a screening level model used to predict osprey exposure to APIs in Chesapeake Bay and evaluated whether exposure levels could have been predicted in Delaware Bay had we just measured concentrations in water or fish. Use of surface water and BCFs did not predict API concentrations in fish well, likely due to fish movement patterns, and partitioning and bioaccumulation uncertainties associated with these ionizable chemicals. Input of highest measured API concentration in fish plasma combined with pharmacokinetic data accurately predicted that diclofenac and acetaminophen would be the APIs most likely detected in osprey plasma. For the majority of APIs modeled, levels were not predicted to exceed 1 ng/mL or method detection limits in osprey plasma. Based on the target analytes examined, there is little evidence that APIs represent a significant risk to ospreys nesting in Delaware Bay. If an API is present in fish orders of magnitude below HTC, sampling of fish-eating birds is unlikely to be necessary. However, several human pharmaceuticals accumulated in fish plasma within a recommended safety factor for HTC. It is now important to expand the scope of diet-based API exposure modeling to include alternative exposure pathways (e.g., uptake from landfills, dumps and wastewater treatment plants) and geographic locations (developing countries) where API contamination of the environment may represent greater risk.

Bioaccumulation of human pharmaceuticals in fish across habitats of a tidally influenced urban bayou.

Though pharmaceuticals and other contaminants of emerging concern are increasingly observed in inland water bodies, the occurrence and bioaccumulation of pharmaceuticals in estuaries and coastal ecosystems are poorly understood. In the present study, bioaccumulation of select pharmaceuticals and other contaminants of emerging concern was examined in fish from Buffalo Bayou, a tidally influenced urban ecosystem that receives effluent from a major (∼200 million gallons per day) municipal wastewater treatment plant in Houston, Texas, USA. Using isotope dilution liquid chromatography-tandem mass spectrometry, various target analytes were observed in effluent, surface water, and multiple fish species. The trophic position of each species was determined using stable isotope analysis. Fish tissue levels of diphenhydramine, which represented the only pharmaceutical detected in all fish species, did not significantly differ between freshwater and marine fish predominantly inhabiting benthic habitats; however, saltwater fish with pelagic habitat preferences significantly accumulated diphenhydramine to the highest levels observed in the present study. Consistent with previous observations from an effluent-dependent freshwater river, diphenhydramine did not display trophic magnification, which suggests site-specific, pH-influenced inhalational uptake to a greater extent than dietary exposure in this tidally influenced urban ecosystem. The findings highlight the importance of understanding differential bioaccumulation and risks of ionizable contaminants of emerging concern in habitats of urbanizing coastal systems.

Determination of nicotine and its metabolites accumulated in fish tissue using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry.

The determination of nicotine and its major metabolites (cotinine and anabasine) in fish tissue was performed using liquid chromatography and tandem mass spectrometry. Marine and freshwater fish were purchased from local grocery stores and were prepared based on a quick, easy, cheap, effective, rugged, and safe sample preparation protocol. To determine the highly polar compounds, hydrophilic interaction liquid chromatography was also used. There were modest suppressions on measured nicotine signals (10%) due to the matrix effects from marine fish but no obvious effects on freshwater fish signals. Method validation was incorporated with internal standards and carried out with matrix-matched calibration. The detection limits for nicotine, cotinine, and anabasine were 9.4, 3.0, and 1.5 ng/g in fish, respectively. Precision was quite acceptable returning less than 8% RSD at low, medium, and high concentrations. Acceptable and reproducible extraction recoveries (70-120%) of all three compounds were achieved, except for anabasine at low concentration (61%). The method was then applied to define nicotine bioaccumulation in a fathead minnow model, which resulted in rapid uptake with steady state internal tissue levels, reached within 12 h. This developed method offers a fast, easy, and sensitive way to evaluate nicotine and its metabolite residues in fish tissues.