Bee Pollinator Mortality Due to Pesticide-Laden Particulate Matter from Beef Cattle Feedyards.

Wild and managed bees are critical for the stability of trophic webs, angiosperm reproduction, and agricultural productivity. Unfortunately, as many as 40% of crop pollinators are in a steep decline due to habitat loss and exposure to agrochemicals. Pyrethroids, neonicotinoids, and macrocyclic lactones are among the many agrochemicals toxic to pollinating insects that are used extensively in industrial beef cattle feeding operations throughout the world. Fugitive feedyard particulate matter (PM) transports agrochemicals into the surrounding environs. To determine the impact of agrochemical-laden feedyard particulate matter on bee pollinators, we conducted in situ experiments wherein honeybees and mason bees were placed downwind and upwind of feedyards (N = 40). Concurrent, colocated total suspended particulate matter samples contained multiple insecticides and parasiticides including pyrethroids, neonicotinoids, and macrocyclic lactones, in significantly higher concentrations downwind of feedyards (bifenthrin, 8.45 ± 4.92; permethrin, 1032.34 ± 740.76; clothianidin, 3.61 ± 1.48; imidacloprid, 73.32 ± 47.52; thiamethoxam, 5.81 ± 3.16; abamectin, 0.45 ± 0.29; ivermectin, 8.88 ± 5.06 ng/g). Honeybees and mason bees sited downwind of feedyards always experienced higher mortality than those correspondingly sited upwind, and male mason bees experienced significantly higher mortality compared to females when both were sited downwind. Bees occurring downwind of beef cattle feedyards for 1 h are 232-260% more likely to die than those occurring upwind. Thus, agrochemicals used on and emitted from beef cattle feedyards are significant threats to bee pollinators.

Influence of Life-History Parameters on Persistent Organic Pollutant Concentrations in Blubber of Eastern North Pacific Gray Whales (Eschrichtius robustus).

Exposure to persistent organic pollutants (POPs) can significantly impact marine mammal health, reproduction, and fitness. This study addresses a significant 20-year gap in gray whale contaminant monitoring through analysis of POPs in 120 blubber biopsies. The scope of this substantial sample set is noteworthy in its range and diversity with collection between 2003 and 2017 along North America's west coast and across diverse sex, age, and reproductive parameters, including paired mothers and calves. Mean blubber concentrations of polychlorinated biphenyls (∑PCBs), dichlorodiphenyltrichloroethanes (∑DDTs), and chlordanes (∑CHLs) generally decreased since previous reports (1968-1999). This is the first report of polybrominated diphenyl ethers (PBDEs) and select hexachlorocyclohexanes (HCHs) in this species. Statistical modeling of the 19 most frequently detected compounds in this dataset revealed sex-, age-, and reproductive status-related patterns, predominantly attributed to maternal offloading. Mean POP concentrations differed significantly by sex in adults (17 compounds, up to 3-fold higher in males) but not in immatures (all 19 compounds). Mean POP concentrations were significantly greater in adults versus immatures in both males (17 compounds, up to 12-fold) and females (13 compounds, up to 3-fold). POP concentrations were detected with compound-specific patterns in nursing calves, confirming maternal offloading for the first time in this species.

Effects of in vitro exposure of perfluorooctanoic acid and monocrotophos on astroglia SVG p12 cells.

Glia cells provide supportive functions to the central nervous system and can be compromised by environmental contaminants. The primary objective of this study was to characterize the effects of in vitro exposure to perfluorooctanoic acid, a persistent environmental contaminant and/or monocrotophos (MCP), a neurotoxic organophosphate that is rapidly metabolized, to astroglia SVG p12 cells. The endpoints evaluated include cell viability, intracellular glutamate levels as a marker of astrocyte homeostasis function, differential gene expression for selected proteins, which include inflammatory markers (tachykinin), astrocytosis (nestin), S100B, and metabolism enzymes (CYP1A1). The results from cell viability revealed significant differences from the controls at some of the concentrations tested. Also, intracellular glutamate levels were elevated at the 10-µM concentration for perfluorooctanoic acid (PFOA) as well as the 10-µM PFOA/5-µM MCP concentration. Gene expression results at 80-µM PFOA concentration revealed a significant increase in the expression of S100B, tachykinin and CYP1A1. A combination of 10-µM PFOA/20-µM MCP caused a significant decrease in the expression of tachykinin. Gene expression for MCP exposures produced a decrease at the 20-µM MCP concentration. Immunofluorescence results indicated an increase in nestin protein expression for the 20-µM concentration of MCP, which contradicted the gene expression at the same concentration tested. The results indicate that toxicity to glia cells can compromise critical glia functions and could be implicated in neurodegenerative diseases.

Transgenerational effects of developmental exposure to chlorpyrifos-oxon in zebrafish (DANIO RERIO).

The organophosphate chlorpyrifos, and its active metabolite chlorpyrifos-oxon (CPO), have been attributed to a number of neurodevelopmental disorders. It is unclear if the adverse effects associated with developmental exposure to the active CPO persist into adulthood and future generations. The goal of this study was to investigate whether CPO-associated changes in embryo-larval zebrafish (ZF) behavior at the F0 5 dpf were manifest throughout the life of the exposed F0, and are inherited by subsequent generations. For this study, embryos were exposed to chlorpyrifos-oxon at the environmentally relevant concentration of 0.01 µg/L and a high concentration of 50 µg/L starting at 4 hpf to 5 dpf, and then raised to F2. There was a significant decrease in distance traveled with 5 dpf F0 ZF exposed to the 50 µg/L CPO, with alterations in noncholinergic genes CFOS and LINGO, and alterations in global DNA methylation. CPO-related behavioral effects were ameliorated by day 21 through the F1 generation. This trend changed with hyperactive behavior, increase acetylcholine concentration in F2 zebrafish that were exposed to 50 µg/L CPO during the F0 development. There was also an increase in AChE activity and hypermethylation in F2 0.01 µg/L exposure larvae, indicating that even low dose exposures can have transgenerational effects. Results from this study demonstrate that early life stage exposures to CPO can lead to epigenetic changes in neurological activity, which may lead to alterations in response to CPO in future generations. ABSTRACT SUMMARY: This study identified a correlation between CPO exposure during F0 development and significant differences in F2 behavioral, AChE activity and neurotransmitter concentration.

Trophic-Level Interactive Effects of Phosphorus Availability on the Toxicities of Cadmium, Arsenic, and Their Binary Mixture in Media-Exposed Scenedesmus acutus and Media and Dietary-Exposed Daphnia pulex.

Various anthropogenic activities simultaneously alter essential mineral nutrients and contaminant content in the environment. Depending on essential nutrient conditions, the uptake and effects of contaminants in exposed organisms may be altered. The addressing of ecological risk assessment (ERA) of contaminant mixtures has proven difficult. Furthermore, most assessments involving single contaminant exposures do not consider the interaction of essential nutrients on toxicological end points. Hypotheses for toxicological effects of cadmium (Cd), arsenic (As), and their binary mixture (Cd/Asmix) include alteration under varying dietary and media phosphorus (P) conditions. However, interactive effects and effect size (η2) are largely unknown. Here, we investigated the toxicities of Cd-, As-, and Cd/Asmix-treated media and diets on Scenedesmus acutus (a primary producer) and Daphnia pulex (a primary consumer), under varied media and dietary P conditions [low (LP), median (MP), and optimum (COMBO)]. Our results showed significant (p < 0.05) interactive effects and concentration dependent growth inhibition of S. acutus. The toxicity (at day 7) of Cd against S. acutus was 2×, 11×, and 4× that of As in LP, MP, and COMBO conditions, respectively, while the joint toxicity effects of Cd/Asmix were partially additive in LP and COMBO, and synergistic in MP media. Furthermore, acute lethal toxicity (96 h) of Cd in D. pulex was ∼60× that of As, while Cd/Asmix joint toxicity was synergistic. Chronic toxicity (14 d) in D. pulex showed significant (p < 0.05) interaction of As and P-availability on survival, reproduction, and behavior (distance moved, velocity, acceleration and mobility), while Cd and P availability showed significant interactive effect on rotational behavior. Dose response effects of Cd, As, and Cd/Asmix in S. acutus and D. pulex were either monophasic or biphasic under varying nutrient conditions. This study provides empirical evidence of the interactive effects of media/dietary P and toxic metals (Cd, As, and Cd/Asmix) at environmentally relevant concentrations, emphasizing the need for consideration of such interactions during ERA.

Characterization of serotonin following exposure to antibiotics in white-tailed deer.

Developing baseline concentrations of serotonin in healthy white-tailed deer will allow for the development of a biomarker using non-invasive sample tissues in sick animals, for example, non-clinical cases of chronic wasting disease. It will also allow some further insight into whether the use of antibiotics as growth promoters (AGP), such as chlortetracycline, is affecting serotonin concentrations in white-tailed deer. Florfenicol and tulathromycin impacts on serotonin concentration changes were also investigated. An analytical method for the detection and confirmation of serotonin, 5-hydroxytryptamine (5-HT), in white-tailed deer tissues was developed and validated. Serum and urine samples were extracted with acetonitrile. Liquid chromatography separation was attained on a Phenomenex C18 column with a Security Guard ULTRA guard column with gradient elution using a mobile phase of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. This methodology was applied to baseline (control), chlortetracycline (CTC) treated, florfenicol treated and tulathromycin treated white-tailed deer serum and urine samples.

Hypoxia induces an increase in intestinal permeability and pulmonary arterial pressures in neonatal Holstein calves despite feeding the flavonoid rutin.

The purposes of this study were to determine whether the naturally occurring flavonoid quercetin, as its glucorhamnoside rutin, reduces intestinal permeability and susceptibility to hypoxia-induced pulmonary hypertension in neonatal Holstein calves. A 2 × 2 between-subjects factorial design was conducted using Holstein steers (n = 16). Factors included oxygen level (simulated altitude of 4,572 m vs. 975 m) and quercetin supplementation as its glucorhamnoside rutin (4 g of quercetin per day vs. 0 g per day). Two days after arrival (d 0 of study) the calves were blocked by body mass into treatment groups, and treatments were initiated. Pulmonary arterial pressure, echocardiography, and serum concentrations of orally administered lactulose (0.45 g/kg) and mannitol (0.15 g/kg) were measured on d 12, 13, and 14, respectively. Calves were euthanized on d 15 and pulmonary tissues collected for semiquantitative scoring of histological lesions. Data were analyzed using linear regression, generalized estimating equations, and 2-sample proportion tests. Hypoxia, but not rutin, was found to be associated with intestinal permeability. The lactulose-mannitol ratio was 0.54 ± 0.13 (standard error) in hypoxic calves and 0.02 ± 0.13 in normoxic controls. Hypoxia increased mean pulmonary arterial pressure. Calves fed rutin under hypoxic conditions tended to have a lower mean pulmonary arterial pressure (59 ± 7 mmHg) than control calves (80 ± 7 mmHg) but similar pressures under normoxic conditions. Paradoxically, however, a greater proportion of calves fed rutin had histological evidence of pulmonary arteriolar medial hypertrophy and adventitial hyperplasia than did controls. In conclusion, the findings of this study indicate that hypoxia increased intestinal permeability in neonatal calves. The flavonoid quercetin, as its glucorhamnoside rutin, had no protective effect on intestinal permeability, and, although it tended to reduce the severity of hypoxia-induced pulmonary hypertension, a greater proportion of calves fed rutin had histological lesions consistent with pulmonary arteriolar remodeling.

Evaluation of the Chronic Reproductive Toxicity of a Fluorine-Free Firefighting Foam and a Short-Chain Fluorinated Foam to Northern Bobwhite Quail (Colinus virginianus).

The development of fluorine-free firefighting foams has been proposed as a way to reduce the adverse environmental consequences of foams containing per- and poly-fluoroalkyl substances. While there are likely fewer environmental and ecological concerns with these new fluorine-free foams in terms of persistence and bioaccumulation, it is prudent to evaluate the ecotoxicity of these fluorine-free foam products given the absence of data. Oral chronic drinking water exposure studies on adult pairs of northern bobwhite quail (Colinus virginianus) were conducted with a short-chain fluorinated and a fluorine-free foam: Buckeye Platinum Plus C6 and National Foam Avio Green KHC, respectively, at three exposure concentrations (0.01%, 0.1%, and 0.25%). Adults were monitored for survival, growth, and reproductive output; and chicks were monitored for survival and growth. Growth parameters in adult quail were not affected by exposure to the Buckeye or Avio foam. However, liver lipid content was higher in adult males exposed to the Buckeye foam or the Avio foam at the highest exposure concentrations. Chicks were heavier and had higher growth rates after adult exposure to Avio at the highest exposure level (0.25%) and to Buckeye at the two lowest exposure levels but not at the highest exposure level. The two adverse reproductive effects observed from avian exposure to Buckeye were an increased percentage of cracked eggs and earlier arrested embryonic development. Similarly, chronic exposure to Avio also induced earlier arrested embryonic development. These results show that the fluorine-free foams tested did cause toxicity to bobwhite quail, but whether they pose a risk at contaminated sites requires further laboratory and field study and additional exposure data. Environ Toxicol Chem 2024;43:211-221. © 2023 SETAC.

Atmospheric deposition of particulate matter from beef cattle feedlots is a likely contributor of pyrethroid occurrence in isolated wetland sediment: Source apportionment and ecological risk assessment.

Industrial cattle feeding operations (feedlots) have been subject to public scrutiny in recent decades regarding environmental impacts of site runoff and aerial dispersion of agrochemical-laden particulate matter (PM). However, source apportionment of multi-use pesticides is challenging in mixed agricultural settings. Beef cattle on feed and row crop production are heavily concentrated in the Southern Great Plains of North America, where playa wetlands are vulnerable to agrochemical inputs and sedimentation from surrounding land use. In the current study, playa basin sediment (n = 33) was analyzed via UHPLC-MS for 21 agrochemicals spanning eight classes (macrocyclic lactones, neonicotinoids, organophosphates, pyrethroids, triazoles, ß-methoxyacrylates, a carboximide, and phenylpyrazole). Pyrethroids were detected most frequently (75.8% of basins). Sediment pyrethroid concentrations were also significantly correlated (R2 = 0.178, p = 0.007) with feedlot proximity (<1-50 km). Principal component analysis (PCA) of land use metrics extracted three principal components (74.3% of total variance), with principal component regression (PCR) showing the greatest agrochemical occurrence in basins heavily weighted by cropland buffer acreage (≤1 km) and feedlot proximity. Sediment toxicity benchmarks protective of two benthic invertebrates (Hyallela azteca and Chironomus spp.) identified λ-cyhalothrin, fenvalerate, and esfenvalerate as individual compounds exceeding levels of acute (RQ > 0.5) and chronic (RQ > 1) concern in >5% and >50% of cases, respectively. However, additive toxicity of co-occurring pyrethroids represents an acute high risk (RI > 1; median RI; acute = 2.4, chronic = 38.6) to benthic invertebrates in >75% of cases, which may threaten higher-order wetland taxa via bioaccumulation and trophic transfer.

Agrochemical occurrence on colocated wildflowers and wild bees collected near beef cattle feed yards and row crops.

It is well established that agrochemicals can pose significant threats to native pollinators; however, relatively little is known about pollinator risks associated with agrochemicals that are used on beef cattle feed yards. Recently, feed yard-derived agrochemicals and those from row crop agriculture were quantified on wildflowers growing on the High Plains, USA. To better characterize pollinator risks on the High Plains, we collected colocated wildflowers and foraging bees across three field seasons for analytical determination of residual agrochemicals. Agrochemicals were detected and quantified on the majority of wildflowers (85%) and nearly half of bees (49%). Permethrin was the most frequently detected analyte on wildflowers (32%) and bees (17%). Flower hazard quotients and flower hazard indices were calculated to deterministically evaluate risk to foraging pollinators. Mean flower hazard quotients exceeded one for 5/16 analytes (31%), and flower hazard quotients calculated for 30% of wildflowers were greater than 50. Flower hazard quotients for clothianidin exceeded 400 for 14% of wildflowers, which portends conditions conducive to frequent bee mortalities. Flower hazard indices were greater on wildflowers from mid-July to mid-September as compared with wildflowers collected earlier in the summer, which coincides with row crop planting and increased prevalence of feed yard flies. Hazard quotients and hazard index values calculated from agrochemical residue data suggest that pollinators frequenting wildflowers near beef cattle feed yards and row crops on the High Plains are at risk from both individual sources, and more so when considered in combination. Integr Environ Assess Manag 2022;18:163-173. © 2021 SETAC.

Acute Oral Toxicity of Nonfluorinated Fire-Fighting Foams to Northern Bobwhite Quail (Colinus virginianus).

Long-chain per- and poly-fluoroalkyl substances (PFAS) have been the active ingredients in firefighting foams for more than 50 years. Due to their extreme persistence, regulatory agencies are concerned about their potential adverse environmental and health impacts. Recently, nonfluorinated chemical constituents have been proposed for use in fire-fighting foams in an effort to reduce the potential negative impacts of PFAS on terrestrial and aquatic flora and fauna. However, it is important to also determine the potential ecotoxicity of these nonfluorinated foam products, because we have little toxicological information for many of them. In preparation for a chronic study, we conducted an acute (24-h) oral toxicity test in northern bobwhite quail (Colinus virginianus) using six different fluorine-free foams; five were commercial foams (BioEx ECOPOL A, Fomtec Enviro USP, National Foam Avio Green KHC, National Foam NFD 20-391, and Solberg Re-Healing Foam), and one was an experimental foam (NRL 502W). A short-chain PFAS-based foam (Buckeye Platinum Plus C6) was also evaluated for comparative purposes. Groups of five birds were initially pseudogavaged with a volume of each product corresponding to a "limit" (the highest exposure concentration expected to occur environmentally). Only one bird (1 of 35) died during the limit test, indicating that all seven products have an acute median lethal dose in adult quail at or above the limit (~1500 mg/kg body wt). Environ Toxicol Chem 2022;41:2003-2007. © 2022 SETAC.

Species- and Tissue-Specific Chronic Toxicity Values for Northern Bobwhite Quail (Colinus virginianus) Exposed to Perfluorohexane Sulfonic Acid and a Binary Mixture of Perfluorooctane Sulfonic Acid and Perfluorohexane Sulfonic Acid.

Per- and polyfluoroalkyl substances (PFAS) are globally distributed and present in nearly every environmental compartment. Characterizing the chronic toxicity of individual PFAS compounds and mixtures is necessary because many have been reported to cause adverse health effects. To derive toxicity reference values (TRVs) and conduct ecotoxicological risk assessments (ERAs) of PFAS-contaminated ecosystems for wildlife, species-specific PFAS chronic toxicity values (CTVs) are needed. The present study quantified PFAS residues from liver and eggs of birds chronically exposed to perfluorohexanoic acid (PFHxA) or a mixture of perfluorooctane sulfonate (PFOS) and PFHxA that produced a no-observable-adverse-effect level (NOAEL) and/or a lowest-observable-adverse-effectlevel (LOAEL). The CTVs we present are lower than those previously reported for birds and should be considered in future regulatory evaluations. From the estimated species- and tissue-specific PFAS CTVs, we found that PFOS and perfluorohexane sulfonate (PFHxS) were more bioaccumulative than PFHxA in avian tissues, but PFHxA was more toxic to reproducing birds than either PFOS or a PFOS:PFHxS mixture. We further determined that avian toxicity was not necessarily additive with respect to PFAS mixtures, which could have implications for PFAS ERAs. The PFAS LOAEL CTVs can be used to predict reproductive and possible population-level adverse health effects in wild avian receptors. Environ Toxicol Chem 2022;41:219-229. © 2021 SETAC.

The Effects of Soil Organic Carbon Content on Plant Uptake of Soil Perfluoro Alkyl Acids (PFAAs) and the Potential Regulatory Implications.

Perfluoro alkyl acids (PFAAs) are known to bioconcentrate in plants grown in contaminated soils; the potential risk from consuming these plants is currently less understood. We determined that the current daily reference doses (RfDs) of the US Environmental Protection Agency (USEPA) could be met by consuming a single radish grown in soils with a perfluorooctanoic acid (PFOA) concentration of 9.7 ng/g or a perfluorooctane sulfonate (PFOS) concentration of 90.5 ng/g. Using a combination of our own research and literature data on plant uptake of PFAAs from soil, we developed equations for predicting PFAA bioconcentration factors (BCFs) for plant shoot and root tissues grown in soils with a known percentage of organic carbon. This calculated BCF was then applied to 6 scenarios with measured soil PFAA concentrations to estimate PFAA concentrations in plants and potential exposure to humans and animals consuming harvested vegetation. Five of the 6 scenarios showed potential for surpassing USEPA PFAA RfDs at soil concentrations as low as 24 ng/g PFOA and 28 ng/g PFOS. Environ Toxicol Chem 2021;40:832-845. © 2020 SETAC.

Emerging and Historical Contaminants Detected in Desert Rodents Collected Near a Low-Level Radioactive Waste Site.

In an effort to determine contaminant presence, concentrations, and movement from a low-level radioactive waste (LLRW) burial disposal site to ecosystems in the surrounding area, a study was developed to assess concentrations of per- and polyfluoroalkyl substances (PFAS), polychlorinated biphenyls (PCBs), and tritium. To complete this assessment small mammals, vegetation, soil, and insect samples were collected from areas within and adjacent to the Beatty, Nevada, LLRW site and from a reference area located approximately 3 km south of the LLRW site. Samples underwent analysis via liquid chromatography tandem mass spectrometry, gas chromatography mass spectrometry, or scintillation spectroscopy depending on the analyte of interest. Small mammal tissues showed maximum concentrations of over 1700 ng/g for PFAS, 1600 ng/g for PCBs, and 10 000 Bq/kg for tritium. The primary contaminants found in soil samples were PCBs, with maximum concentrations exceeding 25 ng/g. Trace amounts of PFAS were also detected in soils and insects. Only qualitative data were obtained from vegetation samples because of the complex matrix of the dominant plant species (creosote bush; Larrea tridentata [Sessé & Moc. ex DC.] Coville). Overall, these data indicate the presence of various anthropogenic contaminants in the ecosystem surrounding the LLRW area, but additional analyses are necessary to confirm the sources and migration pathways of PFAS and PCBs in this hyperarid environment. Environ Toxicol Chem 2021;40:727-734. © 2020 SETAC.

Species- and Tissue-Specific Avian Chronic Toxicity Values for Perfluorooctane Sulfonate (PFOS) and a Binary Mixture of PFOS and Perfluorohexane Sulfonate.

To further characterize avian toxicity to environmental levels of select per- and poly-fluoroalkyl substances (PFAS), we established species- and tissue-specific PFAS chronic toxicity values (CTVs) associated with a lowest-observable-adverse effect level (LOAEL) threshold previously established for northern bobwhite quail (Colinus virginianus) chronically orally exposed via drinking water to either perfluorooctane sulfonate (PFOS) or a simple PFAS mixture. Aided by advances in analytical techniques, the novel avian oral PFAS CTVs reported in the present study are lower than the previously reported toxicity reference values (TRVs) estimated for birds chronically exposed via feed. Thus, current avian PFOS TRVs may not be fully protective of wild avian populations at PFAS-impacted sites. Also, likely due to differences in bioavailability, bioaccessibility, and toxicokinetics among individual PFAS between oral exposure types, we found higher bioaccumulation factors in all assessed tissues from birds exposed via water versus feed. Thus, we propose that future characterization of chemical toxicity due to ingestion exposure initially include a full examination of all probable sources of oral exposure for the most accurate derivation of TRVs and a more complete picture of ecological risk. The avian PFAS LOAEL CTVs established in the present study can be modified with the use of uncertainty factors to derive site-specific avian TRVs for ecological risk assessment at PFAS-impacted sites. From differences observed in the behavior of PFOS when administered as either a single chemical or part of a binary mixture with perfluorohexane sulfonate (PFHxS), we verified that PFOS was absorbed and distributed differently when coadministered with PFHxS and that PFOS likely interacted with PFHxS differently among tissues, helping to explain the differences observed in avian toxicity between exposures. Environ Toxicol Chem 2021;40:899-909. © 2020 SETAC.

Chronic Reproductive Toxicity Thresholds for Northern Bobwhite Quail (Colinus virginianus) Exposed to Perfluorohexanoic Acid (PFHxA) and a Mixture of Perfluorooctane Sulfonic Acid (PFOS) and PFHxA.

Terrestrial toxicology data are limited for comprehensive ecotoxicological risk assessment of ecosystems contaminated by per- and polyfluoroalkyl substances (PFAS) partly because of their existence as mixtures in the environment. This complicates logistical dose-response modeling and establishment of a threshold value characterizing the chronic toxicity of PFAS to ecological receptors. We examined reproduction, growth, and survival endpoints using a combination of hypothesis testing and logistical dose-response modeling of northern bobwhite quail (Colinus virginianus) exposed to perfluorohexanoic acid (PFHxA) alone and to PFHxA in a binary mixture with perfluorooctane sulfonic acid (PFOS) via the drinking water. The exposure concentration chronic toxicity value (CTV) representative of the lowest-observable-adverse effect level (LOAEL) threshold for chronic oral PFAS toxicity (based on reduced offspring weight and growth rate) was 0.10 ng/mL for PFHxA and 0.06 ng/mL for a PFOS:PFHxA (2.7:1) mixture. These estimates corresponded to an adult LOAEL average daily intake CTV of 0.0149 and 0.0082 µg × kg body weight-1 × d-1 , respectively. Neither no-observable-adverse effect level threshold and representative CTVs nor dose-response and predicted effective concentration values could be established for these 2 response variables. The findings indicate that a reaction(s) occurs among the individual PFAS components present in the mixture to alter the potential toxicity, demonstrating that mixture affects avian PFAS toxicity. Thus, chronic oral PFAS toxicity to avian receptors represented as the sum of the individual compound toxicities may not necessarily be the best method for assessing chronic mixture exposure risk at PFAS-contaminated sites. Environ Toxicol Chem 2021;40:2601-2614. © 2021 SETAC.

Nitrate-N-mediated toxicological responses of Scenedesmus acutus and Daphnia pulex to cadmium, arsenic and their binary mixture (Cd/Asmix) at environmentally relevant concentrations.

Several biomarkers used for ecological risk assessment have been established for single contaminant toxicity, many of which are less predictive of the influence of media and/or dietary nutrients on toxicity outcomes of contaminant mixtures. In this study, we investigate toxicological responses and life traits of Scenedesmus acutus and Daphnia pulex to heavy metals (cadmium-Cd, arsenic-As, binary mixture-Cd/Asmix) in media and diets with varied nutrient (nitrate-N) conditions (low-LN, median-MN, optimum-COMBO). Results showed that nitrate-N-mediated metal inhibitory effects on growth and productivity of primary producer (S. acutus) were significantly interactive (p < 0.05; effect size, ƞ2≤56 %). Cadmium toxicities (Cd-IC50s) in S. acutus were 1.2×, 5.3×, and 4.3× As-IC50s in LN, MN and COMBO media, respectively, while mixture (Cd/Asmix) toxicities were synergistic in MN medium and partial additivity in COMBO and LN media. Nitrate-N and metal exposure effects on S. acutus nutrient stoichiometry, metal uptake and bioaccumulation were significantly interactive (p < 0.05, ƞ2≤100 %). Moreover, survival of primary consumer (D. pulex) was significantly impaired by single and mixed dietary-metal exposures with greater effect under LN condition coupled with significant interactive effects on reproductive capacity (p < 0.05, ƞ2≤21.2 %) but not on swimming activity. We recommend that nitrate-N-mediated metal exposure effects/toxicity in bioindicator species should be considered during ecological risk assessments.

Response of exercise-onset vasodilator kinetics to L-citrulline supplementation during different phases of the menstrual cycle.

The purpose of this study was to determine whether L-citrulline (CIT) supplementation during the follicular and luteal phases of the menstrual cycle would present differential effects on vasodilator kinetics in dynamically contracting muscle. Twenty-four women were studied during the follicular (day 15 after onset of menses, n = 13) or the luteal phase (day 25 after onset of menses, n = 11). Supplementation with CIT (6g/day) or placebo occurred 7-days prior to testing in a crossover design across two menstrual cycles. Forearm vascular conductance (FVC) was calculated from blood flow and mean arterial pressure measured continuously during handgrip exercise performed at 10% maximal grip strength. FVC was calculated for each duty cycle (contract:relax, 1:2s) and expressed as a change from baseline (ΔFVC) before being fit with a monoexponential model. Amplitude of the ΔFVC response and the number of duty cycles for ΔFVC to reach 63% of steady-state amplitude (τΔFVC) were derived from the model. Analysis of variance showed no difference in the amplitude of ΔFVC between CIT and placebo (p = .45) or between menstrual cycle phases (p = .11). Additionally, τΔFVC was not different (p = .35) between CIT and placebo in women tested during the follicular (6 ± 3 versus 5 ± 3 duty cycles) or luteal phase (9 ± 1 versus 8 ± 1 duty cycles) although τΔFVC was found to be slower for women tested during the luteal as compared to the follicular phase (8 ± 4 versus 5 ± 3 duty cycles, p = .02). These results indicate that exercise-onset vasodilator kinetics is unaltered with CIT supplementation in young healthy women irrespective of menstrual cycle phase.

Pressurized liquid extraction followed by liquid chromatography coupled to a fluorescence detector and atmospheric pressure chemical ionization mass spectrometry for the determination of benzo(a)pyrene metabolites in liver tissue of an animal model of colon cancer.

Since metabolism is implicated in the carcinogenesis of toxicants, an efficient extraction method together with an analytical method is warranted to quantify tissue burdens of a carcinogen and/or its metabolites. Therefore, the aim of this study was to validate a pressurized liquid extraction (PLE) method for measuring metabolites of benzo(a)pyrene [B(a)P; a food-borne carcinogen] from tissue samples. The sample extraction was performed separately by PLE and liquid-liquid extraction (LLE). PLE followed by high-performance liquid chromatography coupled to online fluorescence detector (HPLC-FLD) was used to quantify separated analytes; and by ultra-high-performance liquid chromatography (UHPLC) coupled to atmospheric pressure chemical ionization tandem mass spectrometry (UHPLC-APCI-MS/MS) were used for confirmation purposes. The UHPLC-MS/MS was set-up in the atmospheric pressure chemical ionization (APCI) positive interface with selective reaction monitoring (SRM). The analytical performance characteristics of the PLE technique was assessed at different temperatures, pressure, number of cycles and solvent types. A methanol + chloroform + water mixture (30:15:10, v/v/v) yielded greater recoveries at an extraction temperature range of 60-80°C, pressure of 10 MPa and an extraction time of 10 min. The PLE method was validated by the analysis of spiked tissue samples and measuring recoveries and limits of quantitation for the analytes of interest using HPLC-FLD equipment. The optimized PLE-HPLC-FLD method was used to quantify the concentrations of B(a)P metabolites in liver samples obtained from a colon cancer animal model. Overall, PLE performed better in terms of extraction efficiency, recovery of B(a)P metabolites and shortened sample preparation time when compared with the classic LLE method.

Chronic Reproductive Toxicity of Perfluorooctane Sulfonic Acid and a Simple Mixture of Perfluorooctane Sulfonic Acid and Perfluorohexane Sulfonic Acid to Northern Bobwhite Quail (Colinus virginianus).

Per- and poly-fluoroalkyl substances (PFAS) are a broad class of environmentally persistent chemicals that include thousands of potentially toxic synthetic organic molecules. Some PFAS have been shown to cause adverse health effects including decreased total cholesterol, birth weight, and reproductive success in laboratory animals; however, a lack of chronic toxicity data exists for PFAS in avian ecological receptors. The present study reports on the chronic toxicity of perfluorooctane sulfonic acid (PFOS) and a mixture of PFOS and perfluorohexane sulfonic acid (PFHxS) to northern bobwhite quail (Colinus virginianus) via oral exposure from drinking water. Female weight gain was adversely affected at an average daily intake (ADI) of 3.10 × 10-3 ± 0.15 × 10-3 mg PFOS:PFHxS (1.2:1) mixture × kg-1 body weight × d-1 . Successful liberation from the shell once pipped was adversely affected at an ADI of 2.45 × 10-3 ± 0.01 × 10-3 mg PFOS x kg-1 body weight × d-1 . These values are comparatively much lower than the current dietary avian toxicity reference value (TRV) derived from birds that were exposed via feed, suggesting the need for updated avian TRVs. Relationships between test chemical (PFOS) and test substance (PFOS:PFHxS) showed that PFOS and PFHxS have possible interacting effects in avian receptors and likely differing mechanisms of toxicity depending on chemical co-occurrence and dose. Both the single-chemical and mixture exposures produced similar and possibly additive toxicity values. Environ Toxicol Chem 2020;39:1101-1111. © 2020 SETAC.