Reproductive success of three passerine species exposed to dioxin-like compounds near Midland, Michigan, USA.

Nests of three passerine birds, house wren (HOWR), tree swallow (TRES), and eastern bluebird (EABL) were monitored daily (2005-2007) at study areas (SAs) downstream of Midland, Michigan where soil and sediment concentrations of polychlorinated dibenzofurans (PCDFs) were significantly greater than the regional background concentrations and upstream reference areas (RAs). Similarly, TRES research conducted at sites contaminated with dioxin-like compounds indicated that concentrations of polychlorinated dibenzo-p-dioxins and PCDFs, expressed as ΣPCDD/DFs and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents observed in the diet and eggs of these three species would be predicted to cause significant effects on reproduction. However, site-specific reproductive parameters including hatching success and fledging success at downstream SAs were similar to or greater than those at upstream RAs. Specifically, hatching success was not significantly different among years, species, locations, or between early and late nesting attempts. Of all initiated clutches, 66% (n = 427), 73% (n = 245), and 64% (n = 122) successfully fledged at least one nestling for HOWR, TRES, and EABL, respectively. Overall reproductive performance was similar between SAs and RAs. The reason for these unexpected results is consistent with the fact that there are species-specific and congener-specific differences in sensitivities to the effects of aryl hydrocarbon receptor agonists.

Dietary exposure of great blue heron (Ardea herodias) to PCDD/DFs in the Tittabawassee River floodplain, MI, USA.

Concentrations of dioxin-like compounds, primarily polychlorinated dibenzofurans (PCDFs), in soils and sediments of the Tittabawassee River (TR) and associated floodplains downstream of Midland, Michigan (USA) were greater than upstream sites and prompted a site-specific risk assessment of great blue herons (GBH). Dietary exposure of GBH to PCDFs and polychlorinated dibenzo-p-dioxins (PCDDs) was evaluated based on site-specific concentrations of residues in prey items. Concentrations of ∑PCDD/DFs and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQ(WHO-Avian)) in prey items collected from the TR were consistently greater than those collected from associated reference areas (RAs) and further downstream in the Saginaw River (SR). The average daily dose (ADD(pot)) of ∑PCDD/DFs to GBH was 45- to 54-fold greater along the TR and 12-fold greater along the SR when compared to the RA. ∑PCDD/DFs were normalized to TEQ(WHO-Avian), and fold differences in the ADD(pot) increased, being 150- to 190-fold greater along the TR and 36-fold greater along the SR than they were in the RA. Greater fold changes in the ADD(pot) based on TEQ(WHO-Avian) between the RA and the TR and SR was due to prey items from the latter reaches having a greater relative toxic potency of ∑PCDD/DFs, primarily from greater amounts of 2,3,7,8-tetrachlorodibenzofuran but also 2,3,4,7,8-pentachlorodibenzofuran. Potential for adverse population-level effects from site-specific contaminant exposures were evaluated via comparison to selected toxicity reference values. The prediction of minimal to no risk of adverse population-level effects resultant from the assessment of site-specific dietary exposure of GBH to ∑PCDD/DFs along the TR and SR is consistent with site-specific assessments of tissue-based exposures as well as population condition.

Sequencing and characterization of mixed function monooxygenase genes CYP1A1 and CYP1A2 of Mink (Mustela vison) to facilitate study of dioxin-like compounds.

As part of an ongoing effort to understand aryl hydrocarbon receptor (AhR) mediated toxicity in mink, cDNAs encoding for CYP1A1 and the CYP1A2 mixed function monooxygenases were cloned and characterized. In addition, the effects of selected dibenzofurans on the expression of these genes and the presence of their respective proteins (P4501A) were investigated, and then correlated with the catalytic activities of these proteins as measured by ethoxyresorufin O-deethylase (EROD) and methoxyresorufin O-deethylase (MROD) activities. The predicted protein sequences for CYP1A1 and CYP1A2 comprise 517 and 512 amino acid residues, respectively. The phylogenetic analysis of the mink CYP1As with protein sequences of other mammals revealed high sequence homology with sea otter, seals and the dog, with amino acid identities ranging from 89 to 95% for CYP1A1 and 81 to 93% for CYP1A2. Since exposure to both 2,3,7,8-Tetrachlorodibenzofuran (TCDF) and 2,3,4,7,8-Pentachlorodibenzofuran (PeCDF) resulted in dose-dependent increases of CYP1A1 mRNA, CYP1A2 mRNA and CYP1A protein levels an underlying AhR-mediated mechanism is suggested. The up-regulation of CYP1A mRNA in liver was more consistent to the sum adipose TEQ concentration than to the liver TEQ concentration in minks treated with TCDF or PeCDF. The result suggested that the hepatic-sequestered fraction of PeCDF was biologically inactive to the induction of CYP1A1 and CYP1A2.

Advanced fluorescence in situ hybridization to localize and quantify gene expression in Japanese medaka (Oryzias latipes) exposed to endocrine-disrupting compounds.

In an earlier study, we described the development of fluorescence in situ hybridization (FISH) using confocal microscopy to localize and quantify gene expression in fish. Here, we report the results of FISH application to investigate effects of model endocrine-disrupting chemicals (EDCs), 17alpha-ethinylestradiol (EE2) and 17beta-trenbolone (TB), on expressions of EDC-responsive genes in Japanese medaka (Oryzias latipes) at the cellular/tissue level paired with histological observation. Gene expressions of vitellogenin-II (Vit-II), androgen receptor (AR), and cytochrome P450 gonadal aromatase (CYP19a) were determined after exposure to 5, 50, or 500 ng/L of EE2 or 50, 500, or 5,000 ng/L of TB for 7 d. Exposure to the greatest concentration of EE2 or TB significantly reduced fecundity and caused histological alterations in gonads. 17alpha-Ethinylestradiol induced Vit-II expression in both male gonads and liver relative to controls and resulted in greater intensity of hematoxylin staining in hepatocytes, which was significantly correlated with Vit-II induction in liver. When exposed to EE2 at less than 50 ng/L, CYP19a expression associated with early stage oocytes was greater than that in controls. However, at 500 ng/L, this trend was reversed. The greater Vit-II expression in testis from all EE2 groups, and the lesser expression of CYP19a in ovaries from the 500 ng/L group, likely is related to changes in the number of cells in which these genes are predominantly expressed rather than to an increase in expression per cell. 17beta-Trenbolone significantly induced AR expression in ovaries but did not alter AR expression in female liver. It was concluded that FISH combined with histology enables advanced elucidation of molecular effects of chemicals by associating changes in gene expression with certain tissues and/or cell types and allows these changes to be related to histological effects.

Perfluoroalkyl acids in marine organisms from Lake Shihwa, Korea.

To our knowledge, this is the first report of concentrations of perfluorooctanesulfonate (PFOS) and other perfluoroalkyl acids (PFAs) in marine organisms from the industrialized region of Korea. Concentrations of eight PFAs were determined in three species of fish (mullet, shad, and rockfish) and three species of marine invertebrates (blue crab, oyster, and mussel) from Lake Shihwa, Korea. This is an area in which relatively great concentrations of PFAs in water and in adjacent industrial effluents have been reported. PFOS was the dominant PFA in marine organisms and most PFOS concentrations were greater than the sum of all other PFAs. The mean concentrations of PFOS were 8.1 x 10 and 3.6 x 10 ng/g, wet weight in liver and blood of fish, respectively. Perfluorocarboxylic acids (PFCAs) were also found in fish, but their concentrations were 10-fold less than those for PFOS. Of the PFCAs measured in fish, concentrations of the longer-chain perfluoroundecanoic acid (PFUnA) were the greatest. Concentrations of PFOS in soft tissues of blue crabs decreased as a function of distance from the shore where inputs from the industrialized areas are discharged into Lake Shihwa. PFOS was the only PFA detectable in mussels and oysters with a mean of 0.5 +/- 0.2 and 1.1 +/- 0.3 ng/g, wet weight, respectively. Concentrations of PFUnA were positively correlated with perfluorodecanoic acid (PFDA) in both the liver and blood of fish, which suggests a common source of these two PFCAs in this area. Hazard quotients developed for fish species were all less than 1.0 for fish collected in Lake Shihwa.

Hepatic P450 enzyme activity, tissue morphology and histology of mink (Mustela vison) exposed to polychlorinated dibenzofurans.

Dose- and time-dependent effects of environmentally relevant concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQ) of 2,3,7,8-tetrachlorodibenzofuran (TCDF), 2,3,4,7,8-pentachlorodibenzofuran (PeCDF), or a mixture of these two congeners on hepatic P450 enzyme activity and tissue morphology, including jaw histology, of adult ranch mink were determined under controlled conditions. Adult female ranch mink were fed either TCDF (0.98, 3.8, or 20 ng TEQ(TCDF)/kg bw/day) or PeCDF (0.62, 2.2, or 9.5 ng TEQ(PeCDF)/kg bw/day), or a mixture of TCDF and PeCDF (4.1 ng TEQ(TCDF)/kg bw/day and 2.8 ng TEQ(PeCDF)/kg bw/day, respectively) for 180 days. Doses used in this study were approximately eight times greater than those reported in a parallel field study. Activities of the cytochrome P450 1A enzymes, ethoxyresorufin O-deethylase (EROD) and methoxyresorufin O-deethylase (MROD) were significantly greater in livers of mink exposed to TCDF, PeCDF, and a mixture of the two congeners; however, there were no significant histological or morphological effects observed. It was determined that EROD and MROD activity can be used as sensitive biomarkers of exposure to PeCDF and TCDF in adult female mink; however, under the conditions of this study, the response of EROD/MROD induction occurred at doses that were less than those required to cause histological or morphological changes.

Fluorescence in situ hybridization techniques (FISH) to detect changes in CYP19a gene expression of Japanese medaka (Oryzias latipes).

The aim of this study was to develop a sensitive in situ hybridization methodology using fluorescence-labeled riboprobes (FISH) that allows for the evaluation of gene expression profiles simultaneously in multiple target tissues of whole fish sections of Japanese medaka (Oryzias latipes). To date FISH methods have been limited in their application due to autofluorescence of tissues, fixatives or other components of the hybridization procedure. An optimized FISH method, based on confocal fluorescence microscopy was developed to reduce the autofluorescence signal. Because of its tissue- and gender-specific expression and relevance in studies of endocrine disruption, gonadal aromatase (CYP19a) was used as a model gene. The in situ hybridization (ISH) system was validated in a test exposure with the aromatase inhibitor fadrozole. The optimized FISH method revealed tissue-specific expression of the CYP19a gene. Furthermore, the assay could differentiate the abundance of CYP19a mRNA among cell types. Expression of CYP19a was primarily associated with early stage oocytes, and expression gradually decreased with increasing maturation. No expression of CYP19a mRNA was observed in other tissues such as brain, liver, or testes. Fadrozole (100 microg/L) caused up-regulation of CYP19a expression, a trend that was confirmed by RT-PCR analysis on excised tissues. In a combination approach with gonad histology, it could be shown that the increase in CYP19a expression as measured by RT-PCR on a whole tissue basis was due to a combination of both increases in numbers of CYP19a-containing cells and an increase in the amount of CYP19a mRNA present in the cells.

Passaic river sediment interstitial water phase I toxicity identification evaluation.

A suite of tests was conducted to evaluate and identify the cause or causes of toxicity in Passaic River sediments. Sediment toxicity was measured with three types of bioassays: a whole sediment bioassay with the marine amphipod, Ampelisca abdita, and interstitial water bioassays with A. abdita and the bioluminescent bacterium Vibrio fisheri (Microtox((R))). In addition, a Phase I Toxicity Identification Evaluation (TIE) was conducted to elucidate the cause of observed toxicity. Analytical concentrations of selected residues in whole sediment and interstitial water from the five sampling stations were considered in conjunction with the conclusions drawn from the toxicity tests and Phase I TIE results. Finally, a toxic units approach was used to evaluate the predicted toxicity of measured interstitial water residue concentrations. There was a lack of toxic response in the short-term interstitial water bioassays, indicating that oxidants, soluble forms of metals, and dissolved phase neutral organics were not likely toxicants. However, there was significant toxicity indicated by the whole sediment A. abidita bioassays. After 10 days, there was complete or near complete mortality in amphipods exposed to all of the sediment samples tested. Removal of interstitial water toxicity by filtration was common to all four stations that exhibited measurable initial toxicity. The observed toxicity characteristics are consistent with particle associated neutral organics. This conclusion is supported by toxicity removal via filtration, lack of toxicity in the Microtox((R)) assays, and the fact that whole sediments were more toxic than was interstitial water.

Exposure and effects assessment of resident mink (Mustela vison) exposed to polychlorinated dibenzofurans and other dioxin-like compounds in the Tittabawassee River basin, Midland, Michigan, USA.

Historically, sediments and floodplain soils of the Tittabawassee River (TR; MI, USA) have been contaminated with polychlorinated dibenzofurans (PCDFs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated biphenyls (PCBs). Median concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) based on 2006 World Health Organization tetrachlorodibenzo-p-dioxin toxic equivalency factors (TEFs) in the diet of mink (Mustela vison) ranged from 6.8 x 10(-1) ng TEQ/kg wet weight upstream of the primary source of PCDF to 3.1 x 10(1) ng TEQ/kg wet weight downstream. Estimates of toxicity reference values (TRVs) derived from laboratory studies with individual PCDDs/PCDFs and PCB congeners or mixtures of those congeners, as well as application of TEFs, were compared to site-specific measures of mink exposure. Hazard quotients based on exposures expressed as concentrations of TEQs in the 95th percentile of the mink diet or liver and the no-observable-adverse-effect TRVs were determined to be 1.7 and 8.6, respectively. The resident mink survey, however, including number of mink present, morphological measures, sex ratios, population age structure, and gross and histological tissue examination, indicated no observable adverse effects. This resulted for multiple reasons: First, the exposure estimate was conservative, and second, the predominantly PCDF congener mixture present in the TR appeared to be less potent than predicted from TEQs based on dose-response comparisons. Given this, there appears to be great uncertainty in comparing the measured concentrations of TEQs at this site to TRVs derived from different congeners or congener mixtures. Based on the lack of negative outcomes for any measurement endpoints examined, including jaw lesions, a sentinel indicator of possible adverse effects, and direct measures of effects on individual mink and their population, it was concluded that current concentrations of PCDDs/PCDFs were not causing adverse effects on resident mink of the TR.

Modulation of steroidogenesis by coastal waters and sewage effluents of Hong Kong, China, using the H295R assay.

BACKGROUND, AIM, AND SCOPE: The presence of a variety of pollutants in the aquatic environment that can potentially interfere with the production of sex steroid hormones in wildlife and humans has been of increasing concern. The aim of the present study was to investigate the effects of extracts from Hong Kong marine waters, and influents and effluents from wastewater treatment plants on steroidogenesis using the H295R cell bioassay. After exposing H295R cells to extracts of water, the expression of four steroidogenic genes and the production of three steroid hormones were measured. MATERIALS AND METHODS: Water samples were collected during the summer of 2005 from 24 coastal marine areas and from the influents and effluents of two major waste water treatment plants (WWTPs) in Hong Kong, China. Samples were extracted by solid phase extraction (SPE). H295R cells were exposed for 48 h to dilutions of these extracts. Modulations of the expression of the steroidogenic genes CYP19, CYP17, 3betaHSD2, and CYP11beta2 were determined by measuring mRNA concentrations by real-time polymerase chain reaction (Q-RT-PCR). Production of the hormones progesterone (P), estradiol (E2), and testosterone (T) was quantified using enzyme linked immunosorbent assays (ELISA). RESULTS: Extracts from samples collected in two fish culture areas inhibited growth and proliferation of H295R cells at concentrations greater or equal to 10(5) L equivalents. The cells were exposed to the equivalent concentration of active substances in 10,000 L of water. Thus, to observe the same level of effect as observed in vitro on aquatic organisms would require a bioaccumulation factor of this same magnitude. None of the other 22 marine samples affected growth of the cells at any dilution tested. Twelve of the marine water samples completely inhibited the expression of CYP19 without affecting E2 production; inhibition of CYP17 expression was observed only in one of the samples while expression of CYP11beta2 was induced as much as five- and ninefold after exposure of cells to extracts from two locations. The expression of the progesterone gene 3betaHSD2 was not affected by any of the samples; only one sample induced approximately fourfold the production of E2. Although more than twofold inductions were observed for P and T production, none of these values were statistically significant to conclude effects on the production of these two hormones. While influents from WWTPs did not affect gene expression, an approximately 30% inhibition in the production of E2 and a 40% increase in P occurred for the exposure with influents from the Sha Tin and Stonecutters WWTPs, respectively. Effluents from WWTPs did not affect the production of any of the studied hormones, but a decrement in the expression of the aldosterone gene CYP11beta2 was observed for the Sha Tin WWTP exposure. No direct correlation could be established between gene expression and hormone production. DISCUSSION: Observed cytotoxicity in the two samples from fish culture areas suggest the presence of toxic compounds; chemical analysis is required for their full identification. Although effluents from WWTPs did not affect hormone production, other types of endocrine activity such as receptor-mediated effects cannot be ruled out. Interactions due to the complexity of the samples and alternative steroidogenic pathways might explain the lack of correlation between gene expression and hormone production results. CONCLUSIONS: Changes observed in gene expression and hormone production suggest the presence in Hong Kong coastal waters of pollutants with endocrine disruption potential and others of significant toxic effects. The aromatase and aldosterone genes seem to be the most affected by the exposures, while E2 and P are the hormones with more significant changes observed. Results also suggest effectiveness in the removing of compounds with endocrine activity by the WWTPs studied, as effluent samples did not significantly affect hormone production. The H295R cell showed to be a valuable toll in the battery required for the analysis of endocrine disrupting activities of complex environmental samples. RECOMMENDATIONS AND PERSPECTIVES: Due to the intrinsic complexity of environmental samples, a combination of analytical tools is required to realistically assess environmental conditions, especially in aquatic systems. In the evaluation of endocrine disrupting activities, the H295R cell bioassay should be used in combination with other genomic, biological, chemical, and hydrological tests to establish viable modes for endocrine disruption and identify compounds responsible for the observed effects.

Plasma to egg conversion factor for evaluating polychlorinated biphenyl and DDT exposures in great horned owls and bald eagles.

The benefits of nondestructive sampling techniques, such as plasma sampling, to directly measure contaminant exposure levels in at-risk or protected raptor populations are many. However, such assays are generally inconsistent with the most certain source of toxicity reference values, which are based on feeding studies and quantified as dietary or "in ovo" (egg-based) concentrations. An accurate conversion factor to translate nondestructive plasma-based contaminant concentrations to comparable egg-based concentrations will prove valuable to risk assessors investigating the potential effects of chemical exposures to raptors. We used databases describing the concentrations of total polychlorinated biphenyls (PCBs) in great horned owls (GHO; Bubo virginianus) and total PCBs and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) in bald eagles (Haliaeetus leucocephalus) from the Great Lakes region (Michigan, Wisconsin, USA) to develop a relationship to predict concentrations of PCBs and DDE in eggs. To develop a robust predictive relationship, all of the source data included concentrations of both total PCBs and/or DDE for nestling blood plasma and egg samples collected from within discrete active nesting territories and, in most instances, the same nest. The key characteristics (slope and elevation) of each relationship were tested for differences related to species and geographic region. Predicted variability of relationships were examined and compared to variability associated with natural systems. The results of statistical testing indicate that applying the conversion factors between species (GHO to bald eagle) and among geographic regions yields predicted egg concentrations that are not statistically dissimilar and are within the natural variability observed for residue concentrations among eggs of raptors within species and region.

Risk assessment of great horned owls (Bubo virginianus) exposed to polychlorinated biphenyls and DDT along the Kalamazoo River, Michigan, USA.

The great horned owl (GHO; Bubo virginianus) was used in a multiple lines of evidence study of polychlorinated biphenyls (PCBs) and p,p'-dichlorodiphenyltrichloroethane (DDT) exposures at the Kalamazoo River Superfund Site (KRSS), Kalamazoo, Michigan, USA. The study examined risks from total PCBs, including 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQWorld Health Organization [WHO]-Avian Toxicity Equivalency Factor [TEF]), and total DDTs (sum of DDT, dichlorodiphenyldichloroethylene [DDE], and dichlorodiphenyldichloroethane [DDD]; sigmaDDT) by measuring concentrations in eggs and nestling blood plasma in two regions of the KRSS (upper, lower) and an upstream reference area (RA). An ecological risk assessment compared concentrations of the contaminants of concern (COCs) in eggs or plasma to toxicity reference values. Productivity and relative abundance measures for KRSS GHOs were compared with other GHO populations. Egg shell thickness was measured to assess effects of p,p'-DDE. The concentrations of PCBs in eggs were as great as 4.7 x 10(2) and 4.0 x 10(4) ng PCB/g, wet weight at the RA and combined KRSS sites, respectively. Egg TEQ(WHO-Avian) calculated from aryl hydrocarbon receptor-active PCB congeners and WHO TEFs ranged to 8.0 and 1.9 x 10(2) pg TEQ(WHO-Avian)/g, (wet wt) at the RA and combined KRSS, respectively. Egg sigmaDDT concentrations were as great as 4.2 x 10(2) and 5.0 x 10(3) ng sigmaDDT/g (wet wt) at the RA and combined KRSS, respectively. Hazard quotients (HQs) for the upper 95% confidence interval (UCI) (geometric mean) and least observable adverse effect concentration (LOAEC) for COCs in eggs were < or = 1.0 for all sites. Hazard quotient values based on the no observable adverse effect concentration (NOAEC) 95% UCI in eggs were < or = 1.0, except at the LKRSS (PCB HQ = 3.1; TEQ(WHO-Avian) HQ = 1.3). Productivity and relative abundance measures indicated no population level effects in the UKRSS.

Effects of perfluorooctane sulfonate on mallard and northern bobwhite quail exposed chronically via the diet.

Adult mallard ducks and northern bobwhite quail were exposed to 0, 10, 50, or 150mg perfluorooctane sulfonate (PFOS)/kg in the diet for up to 21 weeks. Adult health, body and liver weight, feed consumption, gross morphology and histology of body organs, and reproduction were examined. Due to mortality, birds exposed to 50 or 150mg PFOS/kg feed were terminated by Week 7. In quail, the lowest observable adverse effect level (LOAEL) was 10mg PFOS/kg feed based on decreased survivorship of 14-day-old quail offspring. For adult female quail fed 10mg/kg feed, there was a slight but statistically significantly PFOS-related increase in liver weight when compared to controls. When liver weight was normalized to body weight, the statistically significant differences were still observed indicating that PFOS affected liver size. However, no other pathological effects were observed livers of quail from this treatment group which suggests that this enlargement may have been an adaptive response. For adult mallards, no treatment-related effects on feed consumption, body or liver weight, growth, or reproductive performance were observed. There was a slightly greater incidence of small testes (length) in adult male mallards and quail exposed to 10mg PFOS/kg, feed when compared to controls. However, spermatogenesis was not affected and there was no effect on the rates of egg fertilization. Due to transfer to eggs, concentrations of PFOS measured in the liver and blood at study termination were greater in male birds than female birds.

Spatial and temporal trends of poly- and perfluoroalkyl substances in fish fillets and water collected from pool 2 of the Upper Mississippi River.

In 2011, poly- and perfluoroalkyl substances (PFASs) were analyzed in surface water and fish fillet samples taken from Pool 2 of the Upper Mississippi River, a 33-mile stretch inclusive of the Minneapolis/St. Paul, Minnesota (USA) metropolitan area. Approximately 100 each of bluegill, freshwater drum, smallmouth bass, and white bass were sampled within the study area. Surface water samples were also collected from each of the 10 sampling reaches established for the study. Water and fillet samples were analyzed for perfluorinated carboxylic acids (C4-C12), perfluorinated sulfonic acids (C4, C6, and C8), and perfluorooctane sulfonamide. Perfluorooctane sulfonate (PFOS) was observed with the greatest frequency in fish fillets and ranged from 3.0 to 760 ng/g wet weight. Mean (geometric) PFOS concentrations in bluegill, freshwater drum, smallmouth bass, and white bass were 20, 28, 29, and 58 ng/g wet weight, respectively. When compared with fish data collected in 2009, a significant reduction (p < 0.05) in PFOS concentrations was noted. This finding was confirmed based on data from studies conducted in 2012 and 2013. Overall, between 2009 and 2013, PFOS concentrations decreased by 65, 76, and 50% for bluegill, freshwater drum, and white bass, respectively (44% decrease for smallmouth bass from 2009 to 2012). These declines in fish PFOS concentrations are consistent with ongoing efforts to effectively control sources of PFASs to the Mississippi River. Environ Toxicol Chem 2017;36:3138-3147. © 2017 SETAC.

Ecotoxicological evaluation of perfluorooctanesulfonate (PFOS).

Based on available toxicity data, protective screening-level concentrations of PFOS were calculated for aquatic and terrestrial organisms. Using the Great Lakes Initiative, water concentrations of PFOS were calculated to protect aquatic plants and animals. The screening plant value (SPV) protective of aquatic algae and macrophytes was calculated as 2.3 mg PFOS/L. The secondary chronic value protective of aquatic organisms was 1.2 microg PFOS/L. The screening-value water concentrations less than or equal to 1.2 microg PFOS/L would not pose a potential risk to aquatic organisms. Because the aquatic benchmark is based on the most sensitive species, this benchmark should also be protective of other aquatic organisms, including amphibians. The tissue-based TRV for fish was determined to be 87 mg PFOS/kg ww. For terrestrial plants, a screening benchmark was determined to be 1.3 mg PFOS/kg soil ww or 1.5 mg PFOS/kg soil dw, whereas for soil invertebrates such as earthworms the benchmark value was 39 mg PFOS/kg dw soil or 33 mg PFOS/kgww soil. For avian species, dietary, ADI, and egg yolk-based benchmarks were determined as 0.28mg PFOS/kg diet, 0.021mg PFOS/kg bw/d, and 1.7 microg PFOS/mL yolk, respectively. Benchmarks for serum and liver for the protection of avian species were 1.0 microg PFOS/mL and 0.6 microg PFOS/gww, respectively. However, no-effect levels in laboratory studies suggest actual population-level effects would not be expected to occur until a concentration of 6.0mg PFOS/kg in the diet, 5.0 microg PFOS/gww in the liver, or 9.0 microg PFOS/mL in the serum was exceeded, thus indicating the conservative nature of the benchmarks.

Alteration of steroidogenesis in H295R cells by organic sediment contaminants and relationships to other endocrine disrupting effects.

A novel bioassay with the human adrenocortical carcinoma cell line H295R can be used to screen for endocrine disrupting chemicals that affect the expression of genes important in steroidogenesis. This assay was employed to study the effects of organic contaminants associated with the freshwater pond sediments collected in the Ostrava-Karvina region, Czech Republic. The modulation of ten major genes involved in the synthesis of steroid hormones (CYP11A, CYP11B2, CYP17, CYP19, 17betaHSD1, 17betaHSD4, CYP21, 3betaHSD2, HMGR, StAR) after exposure of H295R cells to sediment extracts was investigated using quantitative real-time polymerase chain reaction (PCR). Crude sediment extracts, containing high concentrations of polycyclic aromatic hydrocarbons (PAHs) and moderate amounts of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) significantly stimulated expression of the CYP11B2 gene (up to 10-fold induction), and suppressed expression of 3betaHSD2 and CYP21 genes. A similar pattern was observed with the extracts after treatment with concentrated sulfuric acid to remove labile chemicals (including PAHs) leaving only persistent PCBs, OCPs and potentially PCDD/Fs. Comparison of the results with other mechanistically based bioassays (arylhydrocarbon receptor, AhR, mediated responses in H4IIE-luc cells, and estrogen receptor mediated effects in MVLN cells) revealed significant endocrine disrupting potencies of organic contaminants present in the sediments (most likely antiestrogenicity). Pronounced effects were observed particularly in sediment extracts from the Pilnok Pond which harbors an unusual intersexual population of the narrow-cawed crayfish Pontastacus leptodactylus (Decapoda, Crustacea). This pilot study provided the first experimental evidence of the wider application of the H295R bioassay for screening complex environmental samples, and the results support the hypothesis of chemical-induced endocrine disruption in intersexual crayfish.

Productivity of tree swallows (Tachycineta bicolor) exposed to PCBs at the Kalamazoo River superfund site.

A 123-km stretch of the Kalamazoo River in Michigan, was designated a Superfund site in 1990 due to historical releases of effluent containing polychlorinated biphenyl (PCB)-contaminated paper waste. Risk to bird species in the river ecosystem was evaluated using the tree swallow (Tachycineta bicolor) as a monitor for possible effects due to PCB exposure at two nesting locations, one in the Superfund site and one in an upstream reference location that is less contaminated with PCBs. In 2 of the 3 years of the study, clutch size at the contaminated location was 3.7 +/- 1.4 and 4.8 +/- 0.73 eggs per nest (mean +/- SD), which was significantly less than the clutch size at the reference location (5.0 +/- 1.1 and 5.3 +/- 1.1 eggs per nest). However, there were no statistically significant differences in fledging success, predicted brood size, predicted number of fledglings, or growth of nestlings between the Kalamazoo River Superfund site and an upstream reference location with lesser concentrations of PCBs in the sediments and riparian soils. Productivity and hatching success comparisons between these same sites were also not significantly different; however, the power of these conclusions was less (p < .10). The reduction in clutch size at the co-contaminated location could not be attributed to PCBs due to a number of confounding factors, including Co-cocontaminants, habitat structure, and food availability. Other reproductive parameters were not significantly impaired, and the size of the newly established colony at the Kalamazoo River Superfund site continued to grow over the period of the study. These site-specific observations, combined with multiple lines of evidence approach that considered results reported for the effects of both total PCBs and 2,3,7,8 tetrachlorodibenzo-p-dioxin equivalents (TEQ) on tree swallows at other locations, suggest that there were no significant population-level effects of PCBs on tree swallows at the Kalamazoo River Superfund site.

Tree swallow (Tachycineta bicolor) exposure to polychlorinated biphenyls at the Kalamazoo River superfund site, Michigan, USA.

In 1990, a portion of the Kalamazoo River in Michigan, USA, was designated a Superfund site because of the presence of polychlorinated biphenyls (PCBs) in the sediment and floodplain soils. During a four-year period from 2000 to 2003, several avian species were monitored for reproductive effects and concentrations of PCBs in tissues attributed to food chain transfer from contaminated sediments. The tree swallow (Tachycineta bicolor) was chosen as a model receptor for contamination of passerine species. A top-down methodology was used to evaluate the bioaccumulation of PCBs, including non-ortho and mono-ortho congeners, in tree swallow eggs, nestlings, and adults at the Kalamazoo River area of concern (KRAOC) and at an upstream reference site. Generally, a sixfold difference in tissue concentrations of total PCBs was observed between the two sites with concentrations in eggs and nestlings at the KRAOC ranging from 0.95 to 15 microg PCB/g wet weight. Concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQsWHO Avian) for PCBs, based on bird-specific World Health Organization toxic equivalence factors, were 10- to 30-fold greater in the KRAOC than at the reference location. Egg and nestling TEQsWHO-Avian ranged from 0.21 to 2.4 ng TEQ/g wet weight at the KRAOC. Hazard quotients calculated from literature-derived toxicity reference values were below 1.0 at both the target and the reference site based on the no-observed-adverse-effect level and the lowest-observed-adverse-effect level.

Accumulation of polychlorinated biphenyls from floodplain soils by passerine birds.

Eggs, nestlings, and adults of the eastern bluebird (Sialia sialis) and house wren (Troglodytes aedon) were collected at a polychlorinated biphenyl (PCB)-contaminated site and a reference location on the Kalamazoo River (MI, USA). Eggs and nestlings of eastern bluebirds at the more contaminated location contained concentrations of 8.3 and 1.3 mg/kg, respectively, of total PCBs and 77 and 6.3 ng/kg, respectively, of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs). Eggs, nestlings, and adults of house wrens from the contaminated location contained 6.3, 0.77, and 3.2 mg/kg, respectively, of PCBs and 400, 63, and 110 ng/kg, respectively, of TEQs. Concentrations of total PCBs and TEQs in tissues at the more contaminated location were significantly greater than concentrations in tissues at the reference site for all tissue types of both species. Exposures of the two species studied were different, which suggests that terrestrial-based insectivorous passerine species, foraging in the same area, may have differential exposure to PCBs depending on specific foraging techniques and the insect orders that are targeted. Despite the greater accumulation of PCBs at the more contaminated location, the risk of exposure to PCBs did not exceed the threshold for adverse effects at either location.

Perfluorinated compounds in streams of the Shihwa Industrial Zone and Lake Shihwa, South Korea.

Concentrations of perfluorinated alkyl compounds (PFAs), including perfluorooctane sulfonate (PFOS), perfluorohexanesulfonate, perfluorobutanesulfonate, perfluorooctanesulfonamide, perfluorodecanoate, perfluorononanoic acid, perfluorooctanoate (PFOA), perfluoroheptanoate, and perfluorohexanoate, were measured in the streams of the Shihwa and Banweol industrial areas on the west coast of South Korea as well as the adjacent Lake Shihwa (an artificial lake) and Gyeonggi Bay. Perfluorinated alkyl compounds were concentrated from water using solid-phase extraction and were identified and quantified by liquid chromatography/ triple-quadrapole tandem mass spectrometry. Of the PFAs measured, PFOS and PFOA occurred at the greatest concentrations. Concentrations of PFOS ranged from 2.24 to 651 ng/L, and concentrations of PFOA ranged from 0.9 to 62 ng/L. The concentrations of PFOS observed in Lake Shihwa were among the greatest ever measured in the environment. These results suggest local industrial sources of PFOS and PFOA as well as other PFAs. Because of dilution, the greatest concentrations occur in a rather restricted area, near the points of discharge of the streams that empty into the lake. The greatest measured concentration of PFOS exceeded the threshold for effects predicted for predatory birds consuming aquatic organisms continuously exposed to this level.