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Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants that can be responsible for a variety of deleterious effects on organisms. These adverse outcomes are relatively well studied, but at concentrations rarely found in the environment. Among the documented effects of sublethal acute PAH exposure are reductions in osmoregulatory capacity and immune function, and changes in the function of critical metabolic organs such as the liver. Gulf toadfish (Opsanus beta) were exposed to control seawater (0.006 µg tPAH50/L) or water accommodated fractions of Deepwater Horizon spill oil diluted to 3 flow‐through exposure regimes (0.009, 0.059, and 2.82 µg tPAH50/L) for 7 d, with a recovery period of equal duration. We hypothesized that these chronic exposures would induce the aryl hydrocarbon receptor (AhR)‐mediated pathways and result in significant impacts on markers of osmoregulatory, immune, and metabolic function. We further hypothesized that measurable reversal of these impacts would be observed during the recovery period. Our results indicate that activation of cytochrome P 450 (CYP)1A1 was achieved during exposure and reversed during the recovery phase. The only significant deviations from controls measured were a reduction in plasma glucose in fish exposed to medium and high levels of PAH after 7 d of exposure and a reduction in plasma osmolality fish exposed to high levels of PAHs after 7 d of recovery, when CYP1A1 messenger (m)RNA levels had returned to control levels. Our study illustrates a disconnect between the activation of CYP1A1 in response to environmentally realistic PAHs concentrations and several physiological endpoints and supports the idea that the AhR might not be associated with mediating osmoregulatory, immune, and metabolic changes in Gulf toadfish. Environ Toxicol Chem 2021;40:1075–1086. © 2020 SETAC [ABSTRACT FROM AUTHOR]

Lack of consistent findings in different experimental settings remains a major challenge in toxicogenomics. The present study investigated whether consistency between findings of different microarray experiments can be improved when the analysis is based on a common reference frame ("toxicogenomic universe"), which can be generated using the machine learning algorithm of the self‐organizing map (SOM). This algorithm arranges and clusters genes on a 2‐dimensional grid according to their similarity in expression across all considered data. In the present study, 19 data sets, comprising of 54 different adult fathead minnow liver exposure experiments, were retrieved from Gene Expression Omnibus and used to train a SOM. The resulting toxicogenomic universe aggregates 58 872 probes to 2500 nodes and was used to project, visualize, and compare the fingerprints of these 54 different experiments. For example, we could identify a common pattern, with 14% of significantly regulated nodes in common, in the data sets of an interlaboratory study of ethinylestradiol exposures. Consistency could be improved compared with the 5% total overlap in regulated genes reported before. Furthermore, we could determine a specific and consistent estrogen‐related pattern of differentially expressed nodes and clusters in the toxicogenomic universe by applying additional clustering steps and comparing all obtained fingerprints. Our study shows that the SOM‐based approach is useful for generating comparable toxicogenomic fingerprints and improving consistency between results of different experiments. Environ Toxicol Chem 2020;39:526–537. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC. [ABSTRACT FROM AUTHOR]

The bioaccumulation and biotransformation of the 8:2 fluorotelomer acrylate (CFCHCHOC(O)CH = CH, 8:2 FTAc) was investigated in rainbow trout via dietary exposure. The 8:2 FTAc is a monomer used in the manufacture of fluorinated polymers and has been widely detected in the atmosphere. The parent 8:2 FTAc and suspected intermediate and terminal metabolites were monitored in liver, blood, kidney, bile, and feces during the 5-d uptake and 8-d elimination phases using gas chromatography-mass spectrometry (GC-MS)- and liquid chromatography-tandem mass spectrometry (LC-MS/MS)- based methods. Very low levels of the 8:2 FTAc were detected in the internal tissues and feces, suggesting that the 8:2 FTAc was rapidly biotransformed in the gut or liver. Similarly, low concentrations of the 8:2 fluorotelomer alcohol (FTOH) were accumulated in the fish tissues. The 8:2 saturated fluorotelomer carboxylate (FTCA) was formed in the highest concentration, reaching steady-state tissue concentrations of approximately 1,000 to 1,400 ng/g wet weight. The 8:2 FTUCA and 7:3 FTCA were also accumulated in high levels, at levels approximately 10-fold lower than the 8:2 FTCA. Both the 7:3 FTCA and perfluorooctanoate (PFOA) showed increasing levels throughout the uptake phase and into the initial stages of the elimination phase, indicating continued formation through precursors still present in the body. Perfluorononanoate (PFNA) was formed in low nanogram per gram wet weight levels. The intermediate and terminal metabolites were also detected in the bile and feces, indicating an important elimination pathway for these compounds. In addition, the 8:2 FTOH glucuronide conjugate was measured in relatively high concentrations in the bile and feces. The results of the current study demonstrated a scenario in which a biologically labile compound is biotransformed to terminal metabolites that are much more biologically persistent. Environ. Toxicol. Chem. 2010;29:2726-2735. © 2010 SETAC [ABSTRACT FROM AUTHOR]

Dioxins and related compounds (DRCs) and perfluorinated compounds were measured in the livers of pigs (Sus scrofa) collected from an open waste dumping site in South India. Hepatic concentrations of DRCs and perfluorooctanesulfonate (PFOS; up to 200 ng/g wet wt) were significantly higher in male and female pigs, respectively, collected from the dumping site than in those from a reference site. Results suggest that dumping sites are a source of DRCs and PFOS. Hepatic concentrations of DRCs in piglets were higher than in mothers, especially for the congeners with molecular weights in the range of 360 to 400, implying congener-specific maternal transfer of DRCs in swine. Concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans and some non-ortho dioxin-like polychlorinated biphenyls (PCBs) in the liver of pigs were higher than those in the adipose fat and muscle of the same specimens. In addition, the liver-to-adipose concentration ratios for each congener had a significant positive correlation with the levels of hepatic cytochrome P450 (CYP)1A-like protein, suggesting congener-specific and CYP1A-dependent hepatic sequestration of DRCs in the swine. Total hepatic 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs; 8.9–350 pg/g fat wt) had a significant positive correlation with CYP1A-like protein expression (r = 0.56, p = 0.012), suggesting the induction of CYP1A by DRCs. However, the total TEQs had a significant negative correlation with CYP4A-like protein (r = -0.49, p = 0.029), suggesting repression of peroxisome proliferator-activated receptor-α (PPARα)-mediated signaling pathway by DRCs. Decreases in plasma total thyroxine (T4), free T4, and immunoglobulin (Ig) G were also found in pigs from the dumping site compared with those from the reference site. This study provides insight into the toxicological impacts of DRCs and perfluorinated compounds in wild animals from open waste dumping sites. Environ. Toxicol. Chem. 2010;29:1551–1560. © 2010 SETAC [ABSTRACT FROM AUTHOR]

Abstract-A convenient, simple, and widely used approach for modeling the fate of a chemical in a flowing environmental or biological system is to simulate the system as comprising one or more well-mixed boxes, also known as continuous stirred tank reactors (CSTRs). In principle, any desired level of accuracy can be achieved by increasing the number of boxes. However, highly segmented systems require more input data, they are more computationally intensive, and the results may be more difficult to interpret. Thus there is a tendency to minimize the number of boxes, especially in screening level models. Whereas in the hydrology and engineering literature there is an appreciation of the mathematical errors associated with applying the well-mixed box concept, we believe that these errors are often underappreciated when modeling certain environmental systems. Here, we briefly review the implications of these errors in multimedia models, river and lake simulations, atmospheric transport, flow in soils, gastrointestinal absorption, and metabolism in the liver. The key conclusion is that if over 25% of the chemical entering a box is removed, applying this well-mixed assumption can lead to substantial error. We recommend that results obtained when this criterion is violated be treated with caution. [ABSTRACT FROM AUTHOR]

A bioassay was developed to assess the toxic effects of ingested prey contaminated by polycyclic aromatic hydrocarbons (PAHs) using the teleost Fundulus heteroclitus as a predator and the polychaete Nereis virens as a benthic vector. Ten groups of nine male adult Fundulus were exposed for 21 d to 10 different diets of Nereis contaminated with benzo[a]pyrene (BaP) by spiking dead Nereis with BaP (spiked Nereis [SN] diets, 0-26 μg of BaP per gram dry wt) or by exposing living Nereis to a diet, to sediments, or to both contaminated with BaP (exposed Nereis [EN] diets, 0-16 μg/g dry wt). Another group was exposed to commercial fish food, used as reference diet. Condition and prevalence of histopathological changes were not affected. Exposure to the SN diets containing at least 3.5 μg of BaP per gram dry weight caused an induction of ethoxyresorufin-O-deethylase activity in the intestine but not in the liver. In contrast, fish exposed to the highest doses (≥13.4 μg of BaP per gram dry wt) had increased cellular proliferation rate in the liver but not in the intestine. Quantifiable levels of free BaP tetrol-like metabolites were detected in the bile of fish exposed to diets containing more than 6.8 μg/g dry weight of BaP, and exhibited a dose-response relationship in fish exposed to SN diets. For a similar dose of BaP, EN and SN diets had similar effects. Thus, the BaP metabolic products that could have been produced in Nereis apparently did not contribute to the biomarkers responses. This bioassay can be applied to a variety of prey contaminated naturally or experimentally with PAHs. The present study supports the use of intestinal biomarkers, in addition to hepatic biomarkers, in environmental monitoring to assess the impact of dietary exposure to PAHs. [ABSTRACT FROM AUTHOR]

Juvenile rainbow darters (Etheostoma caeruleum) were exposed to nominal concentrations of 20 to 1,000 ng/L of 17α- ethynylestradiol (EE2) at 120 d posthatch and in a subsequent experiment to 200 ng/L of EE2 with 2.0 to 20,000 ng/L of tamoxifen (TMX) at 150 d posthatch to determine the threshold of estrogen-induced morphological and histological changes in a sexually dimorphic benthic fish species ecologically relevant to southern Minnesota (USA). 17α-Ethynylestradiol induced female-associated urogenital papillae in males at 200 ng/L, enlargement and development of fibrosis in male testes, enlargement of ovary and oocyte size in females, and large fatty inclusions in the liver of both sexes. Exposure to 1,000 ng/L of EE2 caused gross hypertrophy of the liver and kidneys and high mortalities, predominantly in male fish. A low incidence of ovotestes found in all treatment groups was unaffected by EE2, which may be unusual to this species or a response to unknown water contaminants present during the hatching or early development of the darters. Gonadosomatic index was not altered for either sex by any treatment. A TMX level equal to or less than that of EE2 decreased fat accumulation in the liver in both sexes, and a TMX level greater than that of EE2 appeared to prevent urogenital papilla in males. Tamoxifen did not significantly alter fibrosis caused by EE2 in testes. It appears that the presence of TMX in the environment can mask many signs of estrogen exposure, including secondary sexual characteristics, hypertrophy of ovaries and testes, and fatty infiltration of organs. Ovotestes did not prove to be a good indicator of estrogen exposure at this late stage of juvenile darter development. [ABSTRACT FROM AUTHOR]

Chemical pollution of the aquatic environment is almost always the result of multiple rather than single toxic compounds. The possibility of separating the effects of key risk chemicals from those of others, including their joint effects, is of clear theoretical interest and high technical importance. We addressed this goal using multiple gene expression profiling in the liver of juvenile brown trout (Salmo trutta lacustris) exposed to three model chemicals (cadmium, carbon tetrachloride [CCl4], and pyrene) administered singly, in binary and trinary combinations at low acutely sublethal concentrations, and in the partial dose-response manner. Differentially expressed genes were grouped by correlation of profiles, and the dependence on dose was analyzed with multiple regression. Responses to cadmium and CCl4 were largely similar, and no sign of interaction was observed (i.e., in binary combinations, the effects were equal to those produced by the more potent compound, cadmium). Joint effects became apparent in the presence of pyrene, which caused markedly different alterations in gene expression. Using the results of 118 experiments conducted earlier for comparison, we found a group of 23 genes responding to chemical toxicity (cadmium, CCl4, pyrene, and resin acids) with significantly higher probability than that of responding to other stressors (handling or viral and bacterial infections). This group included genes implicated in the immune and stress responses that were markedly enriched in extracellular proteins. In conclusion, we demonstrated that chemical-characteristic genomic endpoints often remain when the chemical is present as part of a binary or a trinary mixture. Despite dissimilar chemistry and different cellular targets, the degree of responses to the combination of cadmium and CCl4 appeared to be less than additive. Chemical interactions or nonadditive effects manifested when a compound with a markedly different mode of action (pyrene) was included into the mixture. [ABSTRACT FROM AUTHOR]

This study investigates reproductive impairment and biochemical changes in fathead minnow (Pimephales promelas) exposed for 39 d to varying concentrations of perfluorooctanoic acid (PFOA) under microcosm conditions. While the concentrations tested in this study were much higher than those normally found in the environment, no mortality was associated with PFOA exposure. Only modest changes were observed in condition factor and in relative liver and gonad size. Significant declines in circulating plasma steroids were observed, but these were accompanied by only limited increases in time to first oviposition and decreases in overall egg production. Peroxisome proliferation, as quantified by fatty acyl-CoA oxidase (FAO) activity, was elevated with low PFOA concentrations but attenuated with exposure to higher PFOA doses. Little evidence was seen of differential induction of peroxisome-associated enzyme activity with sex. Oxidative stress, as quantified by the 2-thiobarbituric acid reactive substances (TBARS) assay, was only modestly influenced by PFOA exposure and is not a significant consequence of FAO activity in fathead minnow. Perfluorooctanoic acid appears to be relatively nontoxic at environmentally relevant concentrations but may impact biochemical and reproductive endpoints under conditions associated with environmental spills. [ABSTRACT FROM AUTHOR]

We evaluated liver and skin tumor prevalence and biomarkers of exposure and response in brown bullhead (Ameiurus nebulosus) from three locations in the Anacostia River (Washington, DC, USA), a Chesapeake Bay region of concern. The Tuckahoe River (Maryland, USA) served as a reference. Each river was sampled in fall 2000 and spring 2001. In the Anacostia, prevalence of liver tumors was 50 to 68%, and prevalence of skin tumors was 13 to 23% in large (≥260 mm, age ≥3 years) bullheads. Liver and skin tumor prevalence was 10 to 17% and 0%, respectively, in small (150-225 mm, age 1-2 years) bullheads. Tuckahoe bullhead liver tumor prevalence was 0 to 3% (large) and 0% (small); none had skin tumors. Biliary polynuclear aromatic hydrocarbon (PAH)-like fluorescent metabolites and liver DNA adduct concentrations were elevated in large and small Anacostia bullheads. Mean adduct concentrations were 16 to 28 times higher than those in Tuckahoe fish. Chromatograms revealed a diagonal radioactive zone, indicating polycyclic aromatic compound (PAC)-DNA adducts. The biomarker data and the 10 to 17% liver tumor prevalence at ages 1 to 2 suggest that these year classes are likely to have a high prevalence as they reach age 3 and older. This study provides the strongest evidence to date of the role of PAHs in tumor development in Anacostia bullheads. [ABSTRACT FROM AUTHOR]

A mercury (Hg) saturation assay for measuring metallothionein (MT) in fish liver was modified by optimizing binding conditions to minimize the mercury and tissue consumed. The revised method uses stable Hg at low concentrations instead of 203Hg. At the reduced Hg concentrations used, MT concentrations in livers homogenized in saline appeared to increase systematically with dilution in both bluegill sunfish (Lepomis macrochirus) and largemouth bass (Micropterus salmoides). This error suggested a binding limitation due to sulfhydryl oxidation or competition for and removal of mercury by non-MT proteins. Homogenizing tissues in trichloroacetic acid (TCA) eliminated the interference. To further evaluate the method, the protocol was tested in the laboratory and field. Metallothionein in bluegill injected with 0.6 mg/kg zinc chloride increased at a rate of 0.03 nmole MT/g liver/h (r² = 0.53, p = 0.001). Linearity improved when data were corrected for protein content (r² = 0.74, p < 0.0001). Metallothionein levels in bluegill from a coal ash--contaminated environment were significantly increased over that of hatchery-reared sunfish (F = 20.17, p = 0.0003). The microscaled procedure minimizes concerns related to radioisotope use and waste generation while retaining the high sensitivity of the 203Hg assay. [ABSTRACT FROM AUTHOR]

In July 2000, 330 individuals of three flatfish species were collected from reference locations and nine sites surrounding the outfall of the Orange County (CA, USA) Sanitation District (OCSD) municipal wastewater discharge. Species included hornyhead turbot (Pleuronichthys verticalis), English sole (Pleuronectes vetulus), and bigmouth sole (Hippoglossina stomata). Livers of sampled animals were examined for the expression of vitellogenin (in males), cytochrome P450 1A (CYP1A), and DNA damage (comet assay). Biliary fluorescent aromatic compounds (FACs) were also measured. Comparisons with tissue and sediment contaminant residues, liver histopathology, and population estimates were assessed to determine whether relationships exist between levels of biological organization. The CYP1A results indicated exposure to planar aromatic hydrocarbons at one nearshore site. Regression analysis of all English sole CYP1A to total sediment polychlorinated biphenyls (PCBs) (r² = 0.97; p < 0.05) indicated a significant correlation. Widespread exposure to estrogens was observed at all sites without correlation to the abundance of the three species examined. Fluorescent aromatic compounds demonstrated no correlation to CYP1A expression. Overall, histopathology and biochemical endpoints did not indicate significant adverse effects in fish at the OCSD outfall. [ABSTRACT FROM AUTHOR]

We studied the estrogenic effects of model chemicals in one-year-old juvenile rainbow trout. Methoxychlor (20 mg/kg), diethylstilbestrol (15 mg/kg), 4-tert-octylphenol (25 and 50 mg/kg), and biochanin A (25 and 50 mg/kg) were injected intraperitoneally on days 1, 4, and 7. Fish were sacrificed on day 9 and examined for multiple biomarkers. All of the test chemicals caused increases in plasma vitellogenin levels, a biomarker of estrogenicity. Treatment with the xenoestrogens decreased hepatic lauric acid hydroxylase activity and, as shown by Western blots, also generally reduced expression of hepatic cytochrome P450s 2K1 (CYP2K1), 2M1 (CYP2M1), and 3A27 (CYP3A27) at the protein level. Both doses of biochanin A also significantly induced P4501A (CYP1A) and greatly increased hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity. These findings suggest that methoxychlor, diethylstilbestrol, 4-tert-octylphenol, and biochanin A were all estrogenic and mimicked 17β-estradiol (E2) in repressing the expression of cytochrome P450 isoforms (CYP2K1, CYP2M1, and CYP3A27) in the rainbow trout liver. Additionally, biochanin A was found to induce CYP1A in this fish species. [ABSTRACT FROM AUTHOR]

Changes in ethoxyresorufin-O-deethylase (EROD) activity were monitored through an extended 6-month dietary exposure to determine the relationship between EROD activity and uptake of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in brook trout, Salvelinus fontinalis. Brook trout were fed labeled TCDD during a 4-week loading phase and an 11-week maintenance phase to achieve whole-body concentrations of 0, 75, 150, 300, 600, and 1,200 pg TCDD/g fish. A spawning phase followed during which no TCDD was introduced. The TCDD had an extended half-life, with maximal levels detected in the late loading-early maintenance phases and 81 d after TCDD had been removed from the diet. Accumulation in liver increased as whole-body target concentration increased but was generally less than half of anticipated whole-body target concentrations. The EROD activity demonstrated a dose-dependent increase. Positive correlations were observed between EROD activity and TCDD body burdens for both males and females. For males, maximal induction was attained early in the maintenance phase and maintained during latter phases. For females, induction was characterized by a biphasic pattern. Maximal induction was attained during late loading--early maintenance, with an attenuated response observed just before spawning. In addition, the induction response was modulated by sex, as induction was lower in females when compared with males. If sexual biases are considered, increased EROD activity may serve as an indicator of level of TCDD exposure and a sublethal predictor of effects of exposure. [ABSTRACT FROM AUTHOR]

Fifty white leghorn chicken (Gallus domesticus) eggs per group were injected with 0.1, 1.0, 10.0, or 20.0 µg perfluorooctane sulfonate (PFOS)/g egg before incubation to investigate the effects of PFOS on the developing embryo. Hatchlings were weighed, examined for gross developmental abnormalities, and transferred to a battery brooder, where they were raised for 7 d. Chicks were then weighed, and 20 birds per treatment were randomly chosen for necropsy. The brain, heart, kidneys, and liver were removed and weighed. Livers were processed further for determination of PFOS concentrations and histological assessment. Hatchability was reduced significantly in all treatment groups in a dose-dependent manner. The calculated median lethal dose was 4.9 µg PFOS/g egg. Perfluorooctane sulfonate did not affect posthatch body or organ weights. Exposure to PFOS caused pathological changes in the liver characterized by bile duct hyperplasia, periportal inflammation, and hepatic cell necrosis at doses as low as 1.0 µg PFOS/g egg. Perfluorooctane sulfonate concentrations in the liver increased in a dose-dependent manner. Based on reduced hatchability, the lowest-observed-adverse-effect level was 0.1 µg PFOS/g egg. [ABSTRACT FROM AUTHOR]

Twelve chemicals were tested for binding affinity to rainbow trout liver estrogen receptor (rbtER) and fathead minnow liver ER (fhmER). The chemicals included estradiol (E2), diethylstilbestrol (DES), ethinylestradiol (EE2), estrone (E1), estriol, tamoxifen (TAM), genistein (GEN), p-nonylphenol (PNP), p-tert-octylphenol (PTOP), methoxychlor (MXC), testosterone, and methyltestosterone (MT). Relative binding affinity (RBA) was calculated for each chemical as a function of E2 binding to the receptor. The estrogens DES, EE2, and E1 bound with high affinity to both receptors, with respective RBAs of 583, 166, and 28% (fathead minnow) and 179, 89, and 5% (rainbow trout). Relative binding affinity of E3, TAM, and GEN for both fhmER and rbtER were moderate, with values between 0.3 and 5%. The alkylphenols had weak affinity for the ERs with RBAs for the fhmER of 0.1 and 0.01 for PNP and PTOP, respectively. Corresponding values for the rbtER are 0.027 and 0.009. Estradiol ([³H]E2) only partially was displaced from both the fhmER and the rbtER by MXC, T, and MT. Comparison of RBAs of the chemicals tested for fhmER and rbtER indicates that the rank order of RBAs essentially are the same for both species. [ABSTRACT FROM AUTHOR]