Growth and reproductive effects from dietary exposure to Aroclor 1268 in mink (Neovison vison), a surrogate model for marine mammals.

Polychlorinated biphenyls (PCBs) from the commercial mixture Aroclor 1268 were historically released into the Turtle-Brunswick River estuary (southeastern Georgia, USA) from industrial operations. Sum PCBs (ΣPCBs) in blubber samples from Turtle-Brunswick River estuary bottlenose dolphins (Tursiops truncatus) have been reported at concentrations more than 10-fold higher than those observed in dolphins from adjacent regional estuaries. Given that toxicity data specific to Aroclor 1268 and applicable to marine mammals are limited, predicting the toxic effects of Aroclor 1268 in dolphins is uncertain, particularly because of its unique congener profile and associated physiochemical characteristics compared with other PCB mixtures. American mink (Neovison vison) were chosen as a surrogate model for cetaceans to develop marine mammalian PCB toxicity benchmarks. Mink are a suitable surrogate species for cetaceans in toxicity studies because of similarities in diet and taxonomic class, and a characteristic sensitivity to PCBs provides a potential safety factor when using mink toxicology data for cross-species extrapolations. Effects of dietary exposure to Aroclor 1268 on reproduction, growth, and mortality in mink were compared with both a negative control and a positive control (3,3',4,4',5-pentachlorobiphenyl, PCB 126). Aroclor 1268 dietary ΣPCB concentrations ranged from 1.8 µg/g feed wet weight to 29 µg/g feed wet weight. Whelp success was unaffected by Aroclor 1268 exposure at any level. Treatment mean litter size, kit growth, and kit survival were adversely affected relative to the negative control at dietary ΣPCB concentrations of 10.6 µg/g feed wet weight and greater.

Enzyme induction and histopathology elucidate aryl hydrocarbon receptor-mediated versus non-aryl hydrocarbon receptor-mediated effects of Aroclor 1268 in American mink (Neovison vison).

Polychlorinated biphenyl (PCB) concentrations reported in preferred prey and blubber of bottlenose dolphins from the Turtle-Brunswick River estuary (Georgia, USA) suggest the potential for adverse effects. However, PCBs in Turtle-Brunswick River estuary dolphins are primarily derived from Aroclor 1268, and predicting toxic effects of Aroclor 1268 is uncertain because of the mixture's unique composition and associated physiochemical characteristics. These differences suggest that toxicity benchmarks for other PCB mixtures may not be relevant to dolphins exposed to Aroclor 1268. American mink (Neovison vison) were used as a surrogate model for cetaceans to characterize mechanisms of action associated with Aroclor 1268 exposure. Mink share similarities in phylogeny and life history with cetaceans and are characteristically sensitive to PCBs, making them an attractive surrogate species for marine mammals in ecotoxicity studies. Adult female mink and a subsequent F1 generation were exposed to Aroclor 1268 through diet, and effects on enzyme induction, histopathology, thyroid hormone regulation, hematology, organ weights, and body condition index were compared to a negative control and a 3,3',4,4',5-pentachlorobiphenyl (PCB 126)-positive control. Aroclor 1268 dietary exposure concentrations ranged from 1.8 µg/g wet weight to 29 µg/g wet weight. Anemia, hypothyroidism, and hepatomegaly were observed in mink exposed to Aroclor 1268 beyond various dietary thresholds. Cytochrome P450 induction and squamous epithelial proliferation jaw lesions were low in Aroclor 1268 treatments relative to the positive control. Differences in enzyme induction and the development of squamous epithelial proliferation jaw lesions between Aroclor 1268 treatments and the positive control, coupled with effects observed in Aroclor 1268 treatments not observed in the positive control, indicate that mechanisms additional to the aryl hydrocarbon receptor-mediated pathway are associated with Aroclor 1268 exposure.

Multiple lines of evidence risk assessment of American robins exposed to polychlorinated dibenzofurans (PCDFS) and polychlorinated dibenzo-P-dioxins (PCDDS) in the Tittabawassee River floodplain, Midland, Michigan, USA.

Concentrations of polychlorinated dibenzofurans (PCDFs) and polychlorinated dibenzo-p-dioxins (PCDDs) in Tittabawassee River floodplain soils and biota downstream of Midland, Michigan, USA, are greater than regional background concentrations. From 2005 to 2008, a multiple lines of evidence approach was utilized to evaluate the potential for effects of PCDD/DFs on American robins (Turdus migratorius) breeding in the floodplains. A dietary-based assessment indicated there was potential for adverse effects for American robins predicted to have the greatest exposures. Conversely, a tissue-based risk assessment based on site-specific PCDD/DF concentrations in American robin eggs indicated minimal potential for adverse effects. An assessment based on reproductive endpoints indicated that measures of hatch success in study areas were significantly less than those of reference areas. However, there was no dose-response relationship between that endpoint and concentrations of PCDD/DF. Although dietary-based exposure and reproductive endpoint assessments predicted potential for adverse effects to resident American robins, the tissue-based assessment indicates minimal to no potential for adverse effects, which is reinforced by the fact the response was not dose related. It is likely that the dietary assessment is overly conservative given the inherent uncertainties of estimating dietary exposure relative to direct tissue-based assessment measures. Based on the available data, it can be concluded that exposure to PCDD/DFs in the Tittabawassee River floodplain would not likely result in adverse population-level effects to American robins.

Dietary- and tissue-based exposure of belted kingfisher to PCDFs and PCDDs in the Tittabawassee River floodplain, Midland, MI, USA.

Concentrations of polychlorinated dibenzofurans (PCDFs) and other dioxin-like compounds in soils and sediments of the Tittabawassee River and associated floodplains downstream of Midland, Michigan, USA, are greater than upstream sites. As a result of these concentrations, which are some of the greatest ever reported, a site-specific exposure assessment of belted kingfisher breeding in the assessment area was conducted. To reduce the uncertainty associated with predicting exposure from abiotic matrices, concentrations of residues were quantified in site-specific prey items and in eggs and nestlings of belted kingfisher. Dietary exposure, expressed as the potential average daily dose, based on site-specific concentrations of PCDFs, polychlorinated dibenzo-p-dioxins (PCDDs), and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQ(WHO-Avian)) in prey items was consistently greater along the Tittabawassee River than in associated reference areas and further downstream sites in the Saginaw River. Concentrations of PCDD/DFs in eggs and nestlings of belted kingfisher varied among sampling areas, being greater in both eggs and nestlings nesting along the Tittabawassee River compared to those of belted kingfisher from upstream reference areas. Geometric mean concentrations of PCDD/DFs were 130 and 200 ng/kg wet weight in eggs and nestlings of belted kingfisher, respectively. These concentrations are the equivalent of 84 and 95 ng TEQ(WHO-Avian)/kg. Site-specific biomagnification factors for select PCDD/DF congeners ranged from <1.0 to 1.8 in belted kingfisher.

Effects of dietary exposure of mink (Mustela vison) to 2,3,7,8-tetrachlorodibenzo-p-dioxin, 2,3,4,7,8-pentachlorodibenzofuran, and 2,3,7,8-tetrachlorodibenzofuran on reproduction and offspring viability and growth.

This study assessed the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,4,7,8-pentachlorodibenzofuran (PeCDF), and 2,3,7,8-tetrachlorodibenzofuran (TCDF) on the reproductive performance of female mink (Mustela vison) and the viability and growth of their offspring. Nine adult female mink were randomly assigned to one of 13 dietary treatments (one control and four doses each of TCDD, PeCDF, and TCDF [2.1-8.4, 4.0-15 and 5.2-25 ng TCDD toxic equivalents (TEQ)/kg body wt/d]). Diets were fed from two months prior to breeding through weaning of offspring at six weeks of age. At least nine kits per treatment group were maintained on their diets through 27 weeks of age. There were no effects on litter size or viability of offspring. No consistent effects were observed on body mass or relative organ masses of animals at any age. 2,3,7,8-Tetrachlorodibenzo-p-dioxin and PeCDF accumulated in the liver and adipose tissue, but TCDF cleared rapidly. The lack of significant effects on reproduction and offspring viability contrasts with effects reported for mink exposed to environmentally derived PCB mixtures with equivalent TCDD potencies. This suggests that it may be inappropriate to apply toxicity reference values associated with PCB mixtures to animals also exposed to TCDD, PeCDF, or TCDF, and the World Health Organization TCDD toxic equivalency factors for some congeners may not be appropriate for mink.

PBDEs and methoxylated analogues in sediment cores from two Michigan, USA, inland lakes.

Polybrominated diphenyl ethers (PBDEs) have been widely studied in sediments from the North American Great Lakes; however, no studies have been conducted of occurrences of methoxylated (MeO-) PBDEs in abiotic compartments in this region. In the present study, 23 tri- to hepta-PBDEs and 12 MeO-PBDEs were analyzed in dated sediment cores collected from two inland lakes (White Lake and Muskegon Lake) in Michigan, USA. Concentrations of Σ(23) PBDEs ranged from 3.9 × 10(-1) to 2.4 × 10(0) and from 9.8 × 10(-1) to 3.9 × 10(0) ng/g dry weight in White Lake and Muskegon Lake, respectively. The historical trends of tri- to hepta-PBDEs in the two lakes were different, possibly because of different input and remediation histories. The tri- to hepta-PBDE profiles were similar in the two lakes, with BDE-47 as the predominant congener, followed by BDE-99 and BDE-183. A different temporal trend for BDE-183 was found compared with other PBDEs, which is consistent with debromination of high-brominated PBDEs during sedimentation and aging. Methoxylated-PBDEs were detected only in Muskegon Lake (3.6 × 10(-3) to 1.2 × 10(-1) ng/g dry wt). Methoxylated PBDEs showed different temporal trends compared with tri- to hepta-PBDEs. The differences in patterns of concentrations of MeO-PBDEs in the two lakes might be due to different aquatic communities in each lake. The occurrences of MeO-PBDEs could be the major source of hydroxylated-polybrominated diphenyl ethers (OH-PBDEs) observed in organisms collected in these freshwater systems.

Dietary exposure of three passerine species to PCDD/DFs from the Chippewa, Tittabawassee, and Saginaw River floodplains, Midland, Michigan, USA.

Dietary exposure of house wrens (Troglodytes aedon), tree swallows (Tachycineta bicolor), and eastern bluebirds (Sialia sialis) to polychlorinated dibenzofurans (PCDFs) and polychlorinated dibenzo-p-dioxins (PCDDs) near Midland, Michigan (USA) was evaluated based on site-specific data, including concentrations of residues in bolus samples and individual invertebrate orders and dietary compositions by study species. Site-specific dietary compositions for the three species were similar to those reported in the literature, but differed in their relative proportions of some dietary items. Oligocheata (non-depurated) and Brachycera (Diptera) contained the greatest average concentrations of ΣPCDD/DFs of the major site-specific dietary items collected via food web-based sampling. Average ingestion values of ΣPCDD/DFs from site-specific bolus-based and food web-based dietary concentrations for nestlings at study areas (SAs) were 6- to 20-fold and 2- to 9-fold greater than at proximally located reference areas (RAs), respectively. Average ingestion values of total 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQ(WHO - Avian)) from site-specific bolus-based and food web-based dietary concentrations for nestlings at SAs were 31- to 121-fold and 9- to 64-fold greater than at proximally located RAs, respectively. Estimates of ΣPCDD/DFs and TEQ(WHO - Avian) tissue concentrations based on nestling dietary exposures were greater than those measured. Plausible explanations include nestling metabolism of 2,3,7,8-tetrachlorodibenzofuran and assimilation rates of less than the 70% assumed to occur over the nestling growth period. Profiles of the relative concentrations of individual PCDD/DF congeners in samples of invertebrates and bolus at SAs on the Tittabawassee River downstream of the source of contamination were dominated by 1,2,3,4,6,7,8,9-octachlorodibenzo-p-dioxin (22% to 44%) and 2,3,7,8-tetrachlorodibenzofuran (18% to 50%).

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.

Tissue-based risk assessment of great blue heron (Ardea herodias) exposed to PCDD/DF in the Tittabawassee River floodplain, Michigan, 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). Tissue exposure of PCDF and polychlorinated dibenzo-p-dioxins (PCDDs) was assessed in multiple GBH tissue types, including blood plasma of adults and eggs, as well as blood plasma, adipose, liver, and muscle of nestlings. Adult GBH exposure was associated with foraging area and age class, with concentrations of PCDD/DF being greater in blood plasma of adult GBH foraging in the TR compared with those foraging in upstream reference areas and in older birds as compared with their younger cohorts. Concentrations of PCDD/DFs and dioxin-like polychlorinated biphenyls (PCBs) in eggs and nestling tissues of GBH collected from rookeries within the TR floodplain were generally similar among rookeries. Mean concentrations of PCDD/DFs in eggs of GBH ranged from 45 to 67 ng/kg, wet weight for the rookeries studied, with a maximum concentration of 210 ng/kg, wet weight observed. Adipose consistently had the greatest concentration of PCDD/DFs of all tissues collected from nestlings of GBH, ranging from 98 to 430 ng/kg, wet weight. Potential for adverse population-level effects from site-specific contaminant exposures were evaluated by comparison with selected toxicity reference values (TRVs). Minimal risk of adverse population-level effects were predicted when exposures measured in tissues of GBH collected from rookeries within the TR were compared with appropriate TRVs. This prediction is consistent with site-specific measures of population condition, which included clutch size and number of nestlings per successful nest.

Passerine exposure to primarily PCDFs and PCDDs in the river floodplains near Midland, Michigan, USA.

House wren (Troglodytes aedon), tree swallow (Tachycineta bicolor), and eastern bluebird (Sialia sialis) tissues collected in study areas (SAs) downstream of Midland, Michigan (USA) contained concentrations of polychlorinated dibenzofurans (PCDFs) and polychlorinated dibenzo-p-dioxins (PCDDs) greater than in upstream reference areas (RAs) in the region. The sum of concentrations of PCDD/DFs (SigmaPCDD/DFs) in eggs of house wrens and eastern bluebirds from SAs were 4- to 22-fold greater compared to those from RAs, whereas concentrations in tree swallow eggs were similar among areas. Mean concentrations of SigmaPCDD/DFs and sum 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (SigmaTEQs(WHO-Avian)), based on 1998 WHO avian toxic equivalency factors, in house wren and eastern bluebird eggs ranged from 860 (430) to 1500 (910) ng/kg wet weight (ww) and 470 (150) to 1100 (510) ng/kg ww, respectively, at the most contaminated study areas along the Tittabawassee River, whereas mean concentrations in tree swallow eggs ranged from 280 (100) to 760 (280) ng/kg ww among all locations. Concentrations of SigmaPCDD/DFs in nestlings of all studied species at SAs were 3- to 50-fold greater compared to RAs. Mean house wren, tree swallow, and eastern bluebird nestling concentrations of SigmaPCDD/DFs and SigmaTEQs(WHO-Avian) ranged from 350 (140) to 610 (300) ng/kg ww, 360 (240) to 1100 (860) ng/kg ww, and 330 (100) to 1200 (690) ng/kg ww, respectively, at SAs along the Tittabawassee River. Concentrations of SigmaTEQs(WHO-Avian) were positively correlated with SigmaPCDD/DF concentrations in both eggs and nestlings of all species studied. Profiles of relative concentrations of individual congeners were dominated by furan congeners (69-84%), primarily 2,3,7,8-tetrachlorodibenzofuran and 2,3,4,7,8-pentachlorodibenzofuran, for all species at SAs on the Tittabawassee and Saginaw rivers but were dominated by dioxin congeners at upstream RAs.

Risk assessment methodologies for exposure of great horned owls (Bubo virginianus) to PCBs on the Kalamazoo River, Michigan.

Dietary exposures of great horned owls (GHO; Bubo virginianus) to polychlorinated biphenyls (PCBs) in the terrestrial food web at the Kalamazoo River, Michigan, USA, were examined. Average potential daily doses (APDD) in GHO diets were 7- to 10-fold and 3-fold greater at the more contaminated location versus a reference location for site-specific exposures quantified as total PCBs and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQ(WHO-Avian)), respectively. Wetland/aquatic prey contributed significantly to PCB exposure and APDD. Estimates of risk based on comparison of modeled dietary intake (e.g., APDD) to toxicity reference values (TRVs), using a hazard quotient (HQ) methodology, varied between diet composition methods (mass basis vs numeric basis). Mass-basis compositions yielded greater HQs at all sites. Potential risks associated with dietary exposures ("bottom-up" risk assessment methodology) were less than (HQ < 1) benchmarks for effects. This result is consistent with risk estimates based on concentrations in tissues ("top-down" risk assessment methodology), and indicated PCBs posed no significant risk to terrestrial raptor species. Colocated and concurrent studies that evaluated GHO reproductive performance (nestling productivity) and relative abundance were consistent with results of the risk assessment. Measures of risk based on HQs were consistent with direct measures of ecologically relevant endpoints (reproductive fitness). Uncertainty in risk estimates is contributed during the selection of TRVs for effects in GHO based on TEQ(WHO-Avian) because of the absence of species-specific, dose-response thresholds. This evaluation indicated that a multiple-lines-of-evidence approach provided the best estimate of risk.

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.

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.

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.

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.

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.

Differential accumulation of polychlorinated biphenyl congeners in the terrestrial food web of the Kalamazoo River Superfund site, Michigan.

A series of field studies was conducted to determine the bioaccumulation of polychlorinated biphenyl (PCB) congeners in the terrestrial food web of the Kalamazoo River flood plain. Samples included colocated soils, native plants likely to be consumed by wildlife, several taxa of terrestrial invertebrates, small mammals, passerine bird eggs, nestlings, and adults, and great horned owl plasma and eggs. Mean concentrations of total PCBs in samples from the former Trowbridge impoundment were 6.5 mg/kg dry weight for soils and 0.023, 0.13, 1.3, 1.3, 1.6, and 8.2 mg/kg wet weight for plants, small herbivorous mammals, depurated earthworms, shrews, great horned owl eggs, and house wren eggs, respectively. Historical data from the Kalamazoo River have reported Aroclor-equivalent total PCB concentrations in the terrestrial food web; however, the degree of environmental weathering of the parent PCB mixtures was unknown. In this study, earthworms and composite samples of coleoptera exhibited PCB congener patterns that were similar to patterns in colocated soils. However, in plants, less chlorinated PCBs (e.g., mono-, di-, tri-, and tetrachlorinated biphenyls) were predominant, and in small mammals, there was a notable enrichment of PCBs 153, 180, 138, 118, and 99. In general, concentrations of PCBs were lower in most biota than in soil from the Kalamazoo River Area of Concern (KRAOC) although there was a modest biomagnification of PCBs from lower trophic level biota to highertrophic levels. As a consequence of environmental weathering of PCBs in the terrestrial food web of the KRAOC, the relative potency of the PCBs (expressed as mg TEQs/kg PCBs) decreased from soil to most biota. While there was a general trend, as expected, in which concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) increased with total PCBs, this relationship was rather poor (R2 = 0.13). Taken together, these data suggest that the differential accumulation of PCB congeners in the terrestrial food web can be explained by congener-specific differences in bioavailability from soil, exposure pathways, and metabolic potential of each of the food web components.

Comparison of risk assessment methodologies for exposure of mink to PCBs on the Kalamazoo River, Michigan.

Risk assessments are generally based on exposures predicted by use of simple models of accumulation from abiotic compartments or the diet. The use of tissue-specific measurements of residue concentrations in wildlife tissues is more accurate and subject to less uncertainty, but these data are often not available. This report compares the results of two different site-specific approaches for assessing the risk of PCBs to mink residing along the Kalamazoo River, MI. The first approach was based on hepatic concentrations of PCBs and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) of mink. The second approach was based on measured concentrations of both PCBs and TEQs in the diets of mink. For each of these methodologies, assessments were based on no observed adverse effect levels (NOAELs) or concentrations (NOAECs) and lowest observed adverse effect levels (LOAELs) or concentrations (LOAECs). Samples of mink (Mustela vison) and its diet were collected from within the Kalamazoo River Area of Concern (KRAOC) and an upstream reference area in the Fort Custer Recreation Area (FC). Hazard quotient (HQ) values were calculated based on congener-specific concentrations of PCBs or TEQs, several toxicity reference values (TRVs), and several assumed dietary compositions. Mean total hepatic concentrations of PCBs were 2.7 and 2.3 mg PCBs/kg, ww, in mink from the KRAOC and FC, respectively. HQs based on the LOAEC and mean hepatic PCB concentrations ranged from 0.37 to 0.87 at KRAOC and 0.31-0.73 at FC. HQs based on PCBs in the diet ranged from 0.20 to 1.8 at KRAOC and from 0.04 to 0.35 at FC. Dietary HQs were less than 10-fold different than tissue-based HQs.

Polychlorinated dibenzo-p-dioxin and dibenzofuran concentration profiles in sediments and flood-plain soils of the Tittabawassee River, Michigan.

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in sediments and flood-plain soils collected along the Tittabawassee River in Michigan ranged from 102 to 53,600 pg/g, dry wt. Mean PCDD/PCDF concentrations in downstream sediment and soil were from 10- to 20-fold greater than those found at locations upstream of Midland, Michigan. Concentrations of PCDD/PCDF in sediments and flood-plain soils from the Tittabawassee watershed were comparable to those found in industrialized areas such as the Housatonic and lower Passaic Rivers in the U.S. Concentrations of PCDDs/PCDFs in soil and sediment were not correlated with total organic carbon (TOC) in sediments or soils. OCDD and 2,3,7,8-TeCDF were the predominant congeners in sediment/soil collected from locations downstream of Midland, Michigan. Principal component analysis of the PCDD/PCDF congener profile suggested the presence of sources originating from a mixture of chlorophenol and other chlorinated compound production. Mass balance analysis of TCDD equivalents (TCDD-EQs) derived from H4IIE-luc bioassay of sediment extracts and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) estimated from instrumental analysis suggested that PCDDs/PCDFs were the major dioxin-like compounds present in sediments. A significant correlation existed between bioassay-derived TCDD-EQs and instrumentally measured TEQs (r2 = 0.94).