Use of a vial equilibration technique to measure the change in fugacity capacity of avian food and feces samples for 1,2,3,4-tetrachlorobenzene.

A vial equilibration technique was used to estimate the fugacity capacities of food and feces samples for 1,2,3,4-tetrachlorobenzene (TCB). The method was calibrated using different volumes of n-octanol and by comparing the measured and predicted fugacity capacity (Zoct) of n-octanol for TCB. The vial equilibration technique showed linearity with increasing amounts of n-octanol added to the vial. However, the measured Zoct was on average 8.5 times lower than the literature estimate and interpreted to be influenced by co-solvent effects. The ratio of fugacity capacities of food/feces was 2.9 and was consistent with the ratio estimated using Zt calculation methods (4.3) which considers partitioning capacity of both lipids and non-lipid organic matter. These results provide experimental support to the use of lipid equivalent approaches as opposed to lipid normalization when estimating the partition capacity of biological samples containing low lipid contents.

Persistent organic pollutants and mercury in marine biota of the Canadian Arctic: an overview of spatial and temporal trends.

This review summarizes and synthesizes the significant amount of data which was generated on mercury (Hg) and persistent organic pollutants (POPs) in Canadian Arctic marine biota since the first Canadian Arctic Contaminants Assessment Report (CACAR) was published in 1997. This recent body of work has led to a better understanding of the current levels and spatial and temporal trends of contaminants in biota, including the marine food species that northern peoples traditionally consume. Compared to other circumpolar countries, concentrations of many organochlorines (OCs) in Canadian Arctic marine biota are generally lower than in the European Arctic and eastern Greenland but are higher than in Alaska, whereas Hg concentrations are substantially higher in Canada than elsewhere. Spatial coverage of OCs in ringed seals, beluga and seabirds remains a strength of the Arctic contaminant data set for Canada. Concentrations of OCs in marine mammals and seabirds remain fairly consistent across the Canadian Arctic although subtle differences from west to east and south to north are found in the proportions of various chemicals. The most significant development since 1997 is improvement in the temporal trend data sets, thanks to the use of archived tissue samples from the 1970s and 1980s, long-term studies using archeological material, as well as the continuation of sampling. These data cover a range of species and chemicals and also include retrospective studies on new chemicals such as polybrominated diphenyl ethers. There is solid evidence in a few species (beluga, polar bear, blue mussels) that Hg at some locations has significantly increased from pre-industrial times to the present; however, the temporal trends of Hg over the past 20-30 years are inconsistent. Some animal populations exhibited significant increases in Hg whereas others did not. Therefore, it is currently not possible to determine if anthropogenic Hg is generally increasing in Canadian Arctic biota. It is also not yet possible to evaluate whether the recent Hg increases observed in some biota may be due solely to increased anthropogenic inputs or are in part the product of environmental change, e.g., climate warming. Concentrations of most "legacy" OCs (PCBs, DDT, etc.) significantly declined in Canadian Arctic biota from the 1970s to the late 1990s, and today are generally less than half the levels of the 1970s, particularly in seabirds and ringed seals. Chlorobenzenes and endosulfan were among the few OCs to show increases during this period while summation operatorHCH remained relatively constant in most species. A suite of new-use chemicals previously unreported in Arctic biota (e.g., polybrominated diphenyl ethers (PBDEs), short chain chlorinated paraffins (SCCPs), polychlorinated naphthalenes (PCNs), perfluoro-octane sulfonic acid (PFOS) and perfluorocarboxylic acids (PFCAs)) has recently been found, but there is insufficient information to assess species differences, spatial patterns or food web dynamics for these compounds. Concentrations of these new chemicals are generally lower than legacy OCs, but there is concern because some are rapidly increasing in concentration (e.g., PBDEs), while others such as PFOS have unique toxicological properties, and some were not expected to be found in the Arctic because of their supposedly low potential for long-range transport. Continuing temporal monitoring of POPs and Hg in a variety of marine biota must be a priority.

The influence of diet properties and feeding rates on PCB toxicokinetics in the ring dove.

The influence of diet properties and feeding rate on the uptake and elimination of polychlorinated biphenyls (PCBs) was investigated in ring doves (Streptopelia rissoria). Elimination rates of PCBs were determined in birds dosed with an Aroclor mixture (1242:1248:1260, 1:1:1) and allowed to depurate on uncontaminated high-lipid or low-lipid/high-fiber diets for 105 days. Uptake rates for seven additional PCBs not found within the above Aroclor mixtures were studied in the same birds by feeding high- or low-lipid PCB-spiked diets for 15 days. The two diet treatments contributed to differences in feeding rates, fecal egestion rates, and total fat volume of the birds. Uptake rate constants of PCBs were higher for the low-lipid diet group, whereas PCB assimilation efficiencies were similar between the two groups. Whole-body elimination rates of Aroclor PCBs were most strongly influenced by the chlorine substitution pattern of the congeners, such that PCBs containing an open meta-para site on one of the phenyl rings were rapidly cleared from the animal. Whole-body elimination rates for persistent PCBs could only be determined for PCB 28; other congeners exhibited negligible elimination over the depuration period. For the latter compounds, fecal elimination rates were estimated using the excreta/carcass distribution coefficient and excreta production rates. For PCB 28, the fecal elimination rate was similar to the whole-body elimination rate, suggesting that fecal elimination of persistent PCBs dominates whole body elimination of these compounds. Diet treatment effects were less evident for PCB elimination rates compared to uptake rates. Steady-state biomagnification factors were estimated to range from 1.0 to 5.1 for rapidly cleared PCBs and from 27 to 97 for persistent congeners.

Body burdens and tissue concentrations of organochlorines in polar bears (Ursus maritimus) vary during seasonal fasts.

Lipophilic organochlorines (OCs) are ingested by mammals through their foods and are generally stored in adipose tissue depots. For some species, such as polar bears, the size of these depots can fluctuate seasonally by several-fold. However, the effect of these fluctuations on the fate of stored OCs in an animal with such labile lipid depots is unknown. We determined the whole body burden and tissue concentrations of OCs in free-ranging polar bears categorized by age (cubs-of-the-year, yearlings and adults) and sex before and after a fast averaging 56 days. Adipose tissue, plasma, and milk samples were analysed for sum of chlorobenzenes (sigma-ClBzs), hexachlorocyclohexanes (sigma-HCHs), chlordanes (sigma-CHLORs), dichlorodiphenyltrichloroethane compounds (sigma-DDTs) and polychlorinated biphenyls (sigma-PCBs). Decline in body mass during fasting ranged from 0.2 kg/day for cubs-of-the-year to 0.9 kg/day for sub-adult and adult males. Although all bears showed a decline in both lipid and lean mass during fasting, patterns of OC whole body burden changes were not consistent among compounds and bear classes. The burdens of sigma-DDTs declined by 11-50% for most bears during fasting, those of sigma-CHLORs declined by 67% during fasting in sub-adult and adult males but remained constant for all females, indicating male-specific metabolism of sigma-CHLORs. As fat depots became depleted, OC concentrations in the remaining adipose tissue varied; sigma-DDTs and sigma-HCHs declined while those of sigma-CHLORs and sigma-PCBs generally increased. Thus. within a 3-4 month fast, most polar bears were able to significantly rid their adipose tissue of sigma-DDTs and sigma-HCHs. Burdens of sigma-CHLORs (except males), sigma-ClBzs and sigma-PCBs remained constant for all classes of bears, therefore there was no significant excretion or metabolism during the fast of the specific congeners in these compound classes typically found in polar bears. The ratio of plasma/adipose tissue and milk/adipose tissue OC concentrations was the same for before and after the fast indicating that OC concentrations in polar bears are probably at a steady state among various body compartments. Concentrations of sigma-CHLORs and sigma-PCBs in milk almost doubled during the fast. As a consequence of this rise in milk OC concentrations, the whole body concentrations of these compounds increased in nursing cubs. Since developing young may be susceptible to the effects of environmental contaminants, the increased exposure of nursing cubs to OCs during a fast by their mothers is noteworthy.

Persistent organochlorine contaminants and enantiomeric signatures of chiral pollutants in ringed seals (Phoca hispida) collected on the east and west side of the Northwater Polynya, Canadian Arctic.

To examine the influence of diet and age on organochlorine contaminant (OC) concentrations in two closely related ringed seal (Phoca hispida) populations enantiomeric fractions (EFs) of chiral contaminants and stable isotopes of nitrogen (delta15N) and carbon (delta13C) were measured along with OCs in ringed seals collected from the east and west side of the Northwater Polynya. Seals from these two locations were feeding at the same trophic level based on delta15N values in muscle but had slightly different sources of carbon based on delta13C measurements in muscle. After removing the influence of age, sex, and blubber thickness, OC concentrations did not vary between ringed seals from the east and west side of the polynya. SigmaPCB, SigmaDDT, and Sigmachlordane were found to increase with age for both male and female seals. The inclusion of older (>20 years) female seals, which may have a reduced reproductive effort, may influence the relationships in females. Stable isotopes failed to describe OC concentrations in ringed seals suggesting that diet was not a major factor in variation of OC concentrations within this ringed seal population. Cis- and trans-chlordane, oxychlordane, and heptachlor epoxide were all nonracemic in the ringed seal blubber but did not vary with age, sex, or collection site. Alpha-HCH appeared racemic (enantiomeric fraction = 0.50 +/- 0.01) in the seals, although this EF is different than those previously observed in their prey species, and was found to vary significantly with age. EF values in the ringed seals varied considerably from other Arctic marine mammals and seabirds, providing addition evidence that the type(s) and characteristic(s) of the enzymes involved in biotransformation of chiral OCs vary between these organisms.

Bioaccumulation and toxicokinetics of 42 polychlorinated biphenyl congeners in American kestrels (Falco sparverius).

The bioaccumulation and toxicokinetics of 42 polychlorinated biphenyls (PCBs) was determined in male American kestrels exposed to an Aroclor-contaminated diet for 120 d followed by a 348-d depuration period. The birds were housed under ambient outdoor temperatures to permit normal fluctuations in body weight during the study. Whole body PCB clearance, plasma/fat distribution coefficients, and plasma PCB clearance constants were determined for individual PCBs to calibrate a two-compartment rate constant model in order to describe PCB elimination in the birds. Plasma/fat partition coefficients (K(PF)) averaged 0.0060 +/- 0.0001 for all congeners of study, were not dependent on chemical hydrophobicity, and did not change in summer versus winter sacrificed animals. Plasma clearance constants (k'pc) for PCBs were observed to be dependent on both chlorine substitution patterns and congener hydrophobicity. Polychlorinated biphenyl congeners categorized as readily cleared congeners contained vicinal meta-para hydrogen substituents on at least one phenyl ring, while slowly cleared congeners were chlorine hindered at these positions. A general equation was derived to predict plasma clearance constants for all tri- to octachlorobiphenyls based on the presence of an open meta-para site on one of the phenyl rings and from the n-octanol-water partition coefficient of the chemical. The equation was validated by comparing predicted versus measured relative biomagnification factors of PCBs determined in birds at the end of the dosing period. The two-compartment model calibrated for PCB elimination in American kestrels may be used to describe PCB toxicokinetics in wild birds provided that seasonal fluctuations in the fat content of the modeled population is known.

Persistent organic pollutants (POPs) in a small, herbivorous, arctic marine zooplankton (Calanus hyperboreus): trends from April to July and the influence of lipids and trophic transfer.

Samples of Calanus hyperboreus, a herbivorous copepod, were collected (n = 20) between April and July 1998, and water samples (n = 6) were collected in May 1998, in the Northwater Polynya (NOW) to examine persistent organic pollutants (POPs) in a high Arctic marine zooplankton. Lipid content (dry weight) doubled, water content (r2 = 0.88) and delta15N (r2 = 0.54) significantly decreased, and delta13C significantly increased (r2 = 0.30) in the C. hyperboreus over the collection period allowing an examination of the role of these variables in POP dynamics in this small pelagic zooplankton. The rank and concentrations of POP groups in C. hyperboreus over the entire sampling was sum of PCB (30.1 +/- 4.03 ng/g, dry weight) > sum of HCH (11.8 +/- 3.23) > sum of DDT (4.74 +/- 0.74), sum of CHLOR (4.44 +/- 1.0) > sum of CIBz (2.42 +/- 0.18), although these rankings varied considerably over the summer. The alpha- and gamma-HCH and lower chlorinated PCB congeners were the most common POPs in C. hyperboreus. The relationship between bioconcentration factor (BCF) and octanol-water partition coefficient (Kow) observed for the C. hyperboreus was linear and near 1:1 (slope = 0.72) for POPs with a log Kow between 3 and 6 but curvilinear when hydrophobic POPs (log Kow > 6) were included. Concentrations of sum of HCH. Sum of CHLOR and sum of CIBz increased over the sampling period, but no change in sum of PCB or sum of DDT was observed. After removing the effects of time, the variables lipid content, water content, delta15N and delta13C did not describe POP concentrations in C. hyperboreus. These results suggest that hydrophobic POP (log Kow = 3.86.0) concentrations in zooplankton are likely to reflect water concentrations and that POPs do not biomagnify in C. hyperboreus or likely in other small, herbivorous zooplankton.

Hexachlorocyclohexane (HCH) isomers and chiral signatures of alpha-HCH in the Arctic marine food web of the Northwater Polynya.

Concentrations of hexachlorocyclohexane (HCH) isomers (alpha, beta, and gamma) and enantiomer fractions (EFs) of alpha-HCH were determined in the Northwater Polynya Arctic marine food web. Relative food web structure was established using trophic level models based on organic delta 15N values. Concentrations of HCH in the samples collected, including water, sediment, benthic invertebrates (four species), pelagic zooplankton (six species), Arctic cod, seabirds (seven species), and ringed seal, were in the range previously reported for the Canadian Arctic. The relative proportion of the HCH isomers varied across the food web and appeared to be related to the biotransformation capacity of each species. For invertebrates and fish the biomagnification factors (BMFs) of the three isomers were > 1 and the proportion of each isomer and the EFs of alpha-HCH were similar to water, suggesting minimal biotransformation. Seabirds appear to readily metabolize gamma- and alpha-HCH based on low BMFs for these isomers, high proportions of beta-HCH (62-96%), and high EFs (0.65-0.97) for alpha-HCH. The alpha- and beta-HCH isomers appear to be recalcitrant in ringed seals based on BMFs > 1 and near racemic EFs for alpha-HCH. The beta isomer appears to be recalcitrant in all species examined and had an overall food web magnification factor of 3.9. EFs of alpha-HCH and the proportion of beta-HCH in sigma-HCH in the food web were highly correlated (r2 = 0.92) suggesting that EFs were a good indicator of a species capability to biotransform alpha-HCH.

Chlordane components and metabolites in seven species of Arctic seabirds from the Northwater Polynya: relationships with stable isotopes of nitrogen and enantiomeric fractions of chiral components.

The Northwater Polynya (NOW) is a large area of year-round open water found in the high Arctic between Ellesmere Island and Greenland. NOW has high biological productivity compared with other arctic marine areas, and supports large populations of several seabird species. Seven species of seabirds, dovekie (Alle alle, DOVE), thick-billed murre (Uria lomvia, TBMU), black guillemot (Cepphus grylle, BLGU), black-legged kittiwake (Rissa tridactyla, BLKI), ivory gull (Pagophila eburnea, IVGU), glaucous gull (Larus hyperboreus, GLGU) and northern fulmar (Fulmaris glacialis, NOFU) were collected in May and June 1998 to determine chlordane concentrations in liver and fat and to examine species differences, relationships with stable isotopes of nitrogen, and enantiomeric fractions (EFs) of chiral components. sigma CHLOR concentrations varied over an order of magnitude among species, from a low of 176 +/- 19 ng/g (lipid corrected) in TMBU liver to a high of 3190 +/- 656 ng/g (lipid corrected) in NOFU liver. Lipid-corrected concentrations of chlordane did not vary between sex for any species or between fat and liver except for the DOVE, that had fat concentrations that were significantly greater than the liver. delta 15N values described a significant percentage of the variability of concentrations for most chlordane components, although less than what has been reported for whole food chains. Slopes of delta 15N versus concentration of chlordane components and sigma CHLOR were similar with the exception of those which were metabolized (trans-chlordane) or formed through biotransformation (oxychlordane). The relative proportions of chlordane components in seabirds were related to phylogeny; the procellariid (NOFU) had the greatest percentage of oxychlordane (> 70%), followed by the larids (BLKI, IVGU and GLGU; 40-50%) and the alcids (DOVE and BLGU; 10-20%). The exception was TBMU, an alcid, where oxychlordane made up > 40% of its chlordane. EFs of chiral components failed to predict concentration or trophic level, but did identify biotransformation differences between species and chlordane components. TBMU appeared to have a greater capacity to metabolize and eliminate chlordane, based on high proportions of oxychlordane, the highest EFs for oxychlordane and heptachlor epoxide, and a delta 15N-sigma CHLOR value which was well below the relationships developed for all seabird species.

Quantifying maternal and dietary sources of 2,2',4,4',5,5'-hexachlorobiphenyl deposited in eggs of the ring dove (Streptopelia risoria).

The influence of maternal versus dietary sources of polychlorinated biphenyls (PCBs) deposited to eggs of ring doves (Streptopelia risoria) was investigated. Mature birds that lay a clutch of two eggs every 12 to 15 d were fed a diet of pellets spiked with 2,2',4,4',5,5'-hexacholorobiphenyl (unlabelled isotope designated as [12C]PCB153) for 25 d. The birds were subsequently switched to a diet containing isotopically labeled [U-13C12]PCB153 at similar concentrations for an additional 63 d. The toxicokinetics of [12C]PCB153 and [13C]PCB153 were followed in egg and carcass samples with time. Isotopic ratios of PCB153 in yolk lipids were identical to those in carcass lipids after introduction of diet 2, and they did not resemble those of the food at the time of egg formation. The data indicate that the PCB composition of eggs directly reflects the composition of maternal tissues, not the diet, at the time of yolk formation. Egg yolk:carcass lipid concentration ratios for the two isotopes were significantly less than one, averaging 0.33+/-0.02 (mean +/- standard error). Lipid normalized egg:maternal tissue contaminant concentration ratios for persistent organochlorine chemicals in the literature were reviewed for other avian species. The results provide a preliminary data set that suggests egg:maternal tissue contaminant concentration ratios vary among species, particularly among species that invest different quantities of lipids to a clutch of eggs. The results support the hypothesis that dilution of contaminants in egg lipids relative to maternal lipids is related to the reproductive strategy employed by the species.

Influence of chemical and biological factors on trophic transfer of persistent organic pollutants in the northwater polynya marine food web.

Persistent organic pollutants (POPs) and stable isotopes of nitrogen (delta 15N) were measured in zooplankton (6 species), a benthic invertebrate (Anonyx nugax), Arctic cod (Boreogadus saida), seabirds (6 species), and ringed seals (Phoca hispida) collected in 1998 in the Northwater Polynya to examine effects of biological and chemical factors on trophic transfer of POPs in an Arctic marine food web. Strong positive relationships were found between recalcitrant POP concentrations (lipid corrected) and trophic level based on stable isotopes of nitrogen, providing clear evidence of POP biomagnification in Arctic marine food webs. Food web magnification factors (FWMFs), derived from the slope of the POP--trophic level relationship, provided an overall magnification factor for the food web but over and underestimated biomagnification factors (BMFs) based on predator--prey concentrations in poikilotherms (fish) and homeotherms (seabirds and mammals), respectively. Greater biomagnification in homeotherms was attributed to their greater energy requirement and subsequent feeding rates. Within the homeotherms, seabirds had greater BMFs than ringed seals, consistent with greater energy demands in birds. Scavenging from marine mammal carcasses and accumulation in more contaminated winter habitats were considered important variables in seabird BMFs. Metabolic differences between species resulted in lower than expected BMFs, which would not be recognized in whole food web trophic level--POP relationships. The use of sigma POP groups, such as sigma PCB, is problematic because FWMFs and BMFs varied considerably between individual POPs. FWMFs of recalcitrant POPs had a strong positive relationship with log octanol--water partition coefficient (Kow). Results of this study show the utility of using delta 15N to characterize trophic level and trophic transfer of POPs but highlight the effects of species and chemical differences on trophic transfer of POPs that can be overlooked when a single magnification factor is applied to an entire food web.

Monitoring temporal and spatial trends in polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in eggs of great blue heron (Ardea herodias) on the coast of British Columbia, Canada, 1983-1998.

Eggs from 21 resident great blue heron (Ardea herodias) rookeries were monitored from 1983 to 1998 along the coast of British Columbia, Canada, for contamination with polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs). Dominant congeners (1,2,3,7,8-PnCDD, 1,2,3,6,7,8-HxCDD, 2,3,7,8-TCDD, 2,3,7,8-TCDF, 1,2,3,7,8,9-HxCDD and 2,3,4,7,8-PnCDF) fell markedly in the early 1990s after pulp mills changed from molecular chlorine bleaching to alternative bleaching technologies, and the use of chlorophenolic wood preservatives and anti-sapstains was severely restricted. Strong positive linear regressions between prey fish and heron egg contaminant levels suggested that local dietary uptake was an important route of exposure for herons. Toxic equivalent concentrations (TEQs) sufficient to produce embryotoxicity in great blue heron chicks were measured in eggs from 1985 to 1991 at some colonies. Despite reduction in PCDD/Fs, estimated TEQs remained elevated throughout the 1980s at some urban colonies due to contributions from PCBs.

Analysis of hydroxylated metabolites of PCBs (OH-PCBs) and other chlorinated phenolic compounds in whole blood from Canadian inuit.

In this study, we identified the main hydroxylated polychlorinated biphenyls (OH-PCBs) and other chlorinated phenolic compounds and we determined their relative concentrations in whole blood from 13 male and 17 female Inuit from northern Quebec, Canada, and from a pooled whole blood sample from southern Quebec. We also determined concentrations of polychlorinated biphenyls (PCBs). Total OH-PCB concentrations were variable among the Inuit samples, ranging over 2 orders of magnitude (0.117-11.6 ng/g whole blood wet weight). These concentrations were equal to and up to 70 times those found for the southern Quebec pooled whole blood sample. Geometric mean concentrations of total OH-PCBs were 1.73 and 1.01 ng/g whole blood for Inuit men and women, respectively, and 0.161 ng/g whole blood for the southern population pool. There are limited data available for comparison, but the levels of OH-PCBs in Inuit are higher than those previously reported in the literature for other populations. There was a significant correlation (p < 0.005) between OH-PCBs and PCBs (r = 0.84) and both correlated significantly (p < 0.005) with age (r = 0.68 and 0.78, respectively). The ratio of OH-PCBs to PCBs was lower in Inuit (0.11) than in the southern Quebec pool (0.33). There is no apparent explanation for the difference. There was considerable variability in the congener pattern of the identified OH-PCBs. The main metabolite, 4-OH-CB109 (4-OH-2,3,3',4', 5-pentachlorobiphenyl), constituted 12-62% of the total OH-PCBs in the samples. Pentachlorophenol (PCP) was the dominant phenolic compound in blood, constituting 46% (geometric mean) of the total quantitated chlorinated phenolic compounds. PCP concentrations in Inuit blood ranged from 0.558 to 7.77 ng/g on a wet weight basis. All but two Inuit samples had lower concentrations than the southern Quebec pool (6.29 ng/g). The possible role of OH-PCBs in mediating PCB-induced adverse effects needs to be investigated further.

Geographical differences and time trends of persistent organic pollutants in the Arctic.

Geographical and temporal trends of total PCBs (SigmaPCBs) and hexachlorocyclohexanes (SigmaHCHs) have been examined in polar bear (Ursus maritimus) adipose tissue and ringed seal (Phoca hispida) blubber along a transect from Alaska to Svalbard and northern Norway/western Russia. PCB concentrations in polar bear show a strong west to east trend with higher levels in east Greenland, Svalbard. In ringed seal, highest PCB levels were found at two sites near the Russian coast. SigmaHCHs levels in ringed seals and polar bear show the reverse trend coinciding with much higher levels of SigmaHCHs in seawater in the North American Arctic. Some of these geographical differences may reflect dietary differences especially in the case of PCBs. Levels of SigmaPCBs in polar bears in the eastern Canadian Arctic appear to have increased from the 1970s to the 1980s but are now on the decline. SigmaPCBs and SigmaHCHs levels showed no significant change from the mid-1980s to 1990s in ringed seal blubber from three locations in the eastern Canadian Arctic.

Cytotoxicity and aromatase (CYP19) activity modulation by organochlorines in human placental JEG-3 and JAR choriocarcinoma cells.

The human placental JEG-3 and JAR choriocarcinoma cell lines have been used as placental models for the study of aromatase (CYP19) activity and endocrine functions. In the present study, 21 organochlorines (OCs) mediated decreases in aromatase activity and protein and DNA content and increases in the percent lactate dehydrogenase (LDH) leakage in JEG-3 cells. These effects were highly variable among the types of OC and their treatment concentrations. Lowest observed effective concentrations reached 0. 001 microM for several OCs. Aromatase activity decreases and OC-mediated cytotoxicity were related. Thus, it was not possible to clearly assess the capacity of the OCs to modulate aromatase activity. Similar to 1,4-naphthoquinone, the most cytotoxic OCs contained a hydroxyl (4'-OH-2,4,6-trichlorobiphenyl and tris(4-chlorophenyl)methanol) or methylsulfonyl- (3- and 4-MeSO(2)-2, 2',5,5'-tetrachlorobiphenyl and -2,3',4',5-tetrachlorobiphenyl, and 3'- and 4'-MeSO(2)-2,2',3,4,5'-pentachlorobiphenyl and -2,2',4,5, 5'-pentachlorobiphenyl) functional group. Modulation of aromatase activity and LDH leakage were less for 3,3',4,4', 5-pentachlorobiphenyl and benzo[a]pyrene and insignificant for five alkyl-substituted trichloro-dibenzofurans and 2,3,7, 8-tetrachloro-dibenzo-p-dioxin (up to 10 microM). Cytotoxicity-related effects were influenced by the cell density and the presence of 10% fetal calf serum in the medium during compound incubation. Similar cytotoxic effects were observed for the JAR cell line. The involvement of an apoptotic mechanism of cytotoxicity in OC-treated JEG-3 cells was suggested by the binding of APO2.7 (an antibody specific to apoptotic cells), DNA fragmentation, and trypan blue staining. JEG-3 and JAR cells appear too sensitive toward OC-mediated cytotoxicity for use as in vitro bioassays to evaluate the potential modulation of aromatase activity. However, these cell lines may prove useful for examining the capacity of xenobiotics to modulate placental toxicity.

Contaminants in ospreys from the Pacific Northwest: I. Trends and patterns in polychlorinated dibenzo-p-dioxins and -dibenzofurans in eggs and plasma.

Osprey (Pandion haliaetus) eggs were collected from 1991 to 1997 at nests (n = 121) upstream and downstream of bleached kraft pulp mills and at reference sites in the Fraser and Columbia River drainage systems of British Columbia, Washington, and Oregon. Blood samples were collected from nestling ospreys during the 1992 breeding season on the Thompson River. Samples were analyzed for polychlorinated dibenzo-p-dioxins (PCDDs) and -dibenzofurans (PCDFs). Mean concentrations of 2,3,7,8-TCDD were significantly higher in eggs collected in 1991 at downstream compared to upstream nests near pulp mills at Kamloops and Castlegar, British Columbia. There were no significant temporal trends in 2,3,7,8-TCDD, -TCDF or other measured compounds at a sample of nests monitored between 1991 and 1994 downstream of the Castlegar pulp mill, despite changes in bleaching technology (CIO2 substitution). However, by 1997 concentrations of 2, 3,7,8-TCDD and -TCDF were significantly lower than previous years in nests sampled downstream at both Castlegar and Kamloops. An unusual pattern of higher chlorinated PCDDs and PCDFs was found in many of the osprey eggs collected in this study, and considerable individual variation in the pattern existed among eggs from the same site. For example, eggs from four different nests at one study area (Quesnel) on the Fraser River had concentrations of 1,2,3,4,6,7,8-HpCDD ranging from <1 to 1,100 ng/kg and OCDD from <1 to 7,000 ng/kg wet weight. Higher mean concentrations of HpCDD and OCDD were found in eggs from the Thompson River, a tributary of the Fraser, compared to the Columbia River, and concentrations were generally higher at nests upstream of pulp mills. In plasma samples, 1,2,3,4,6,7,8-HpCDD and OCDD were the main compounds detected, with no significant differences measured between samples upstream versus downstream or earlier versus later in the breeding season. Use of chlorophenolic wood preservatives by lumber processors was considered the main source of higher chlorinated PCDD/Fs throughout the systems, based on patterns of trace PCDFs in eggs and significant correlations between egg concentrations of pentachlorophenol and both HpCDD (r = 0.891, p < 0.01) and OCDD (r = 0.870, p < 0.01).

Chlorinated hydrocarbon contaminants in polar bears from eastern Russia, North America, Greenland, and Svalbard: biomonitoring of Arctic pollution.

Adipose tissue samples from polar bears (Ursus maritimus) were obtained by necropsy or biopsy between the spring of 1989 to the spring of 1993 from Wrangel Island in Russia, most of the range of the bear in North America, eastern Greenland, and Svalbard. Samples were divided into 16 regions corresponding as much as possible to known stocks or management zones. Concentrations of dieldrin (DIEL), 4,4'-DDE (DDE), sum of 16 polychlorinated biphenyl congeners (sigma PCB), and sum of 11 chlordane-related compounds and metabolites (sigma CHL) were determined. In order to minimize the effect of age, only data for adults (320 bears age 5 years and older) was used to compare concentrations among regions. Concentrations of sigma PCB were 46% higher in adult males than females, and there was no significant trend with age. Concentrations of sigma CHL were 30% lower in adult males than females. Concentrations of sigma PCB, sigma CHL, and DDE in individual adult female bears were standardized to adult males using factors derived from the least-square means of each sex category, and geometric means of the standardized concentrations on a lipid weight basis were compared among regions. Median geometric mean standardized concentrations (lipid weight basis) and ranges among regions were as follows: sigma PCB, 5,942 (2,763-24,316) micrograms/kg; sigma CHL, 1,952 (727-4,632) micrograms/kg; DDE, 219 (52-560) micrograms/kg; DIEL, 157 (31-335) micrograms/kg. Geometric mean sigma PCB concentrations in bears from Svalbard, East Greenland, and the Arctic Ocean near Prince Patrick Island in Canada were similar (20,256-24,316 micrograms/kg) and significantly higher than most other areas. Atmospheric, oceanic, and ice transport, as well as ecological factors may contribute to these high concentrations of sigma PCB. sigma CHL was more uniformly distributed among regions than the other CHCs. Highest sigma CHL concentrations were found in southeastern Hudson Bay, which also had the highest DDE and DIEL concentrations. In general, concentrations of sigma CHL, DDE, and DIEL were higher in eastern than western regions, suggesting an influence of North American sources. Average sigma PCB concentrations in bears from the Canadian Arctic were similar to those in 1982-84, while average sigma CHL and DDE concentrations were 35-44% lower and DIEL was 90% lower. However, the significance of these temporal trends during the 1980s is not conclusive because of the problems of comparability of data.

Environmental contamination and developmental abnormalities in eggs and hatchlings of the common snapping turtle (Chelydra serpentina serpentina) from the Great Lakes-St Lawrence River basin (1989-1991).

During 1989-1991, we assessed developmental abnormalities in embryos and hatchlings from eggs of the common snapping turtle (Chelydra serpentina serpentina). Eggs were collected and artificially incubated from eight sites in Ontario, Canada and Akwesasne/New York, USA. In eggs from the same clutches we measured 20 organochlorine pesticides, 48 polychlorinated biphenyl (PCBs) congeners including 6 non-ortho PCBs, 8 polychlorinated dibenzodioxins (PCDDs), 14 polychlorinated dibenzofurans (PCDFs) and total mercury. We found a significant increase in abnormal development with increasing polychlorinated aromatic hydrocarbon exposure in eggs, particularly PCDD and PCDF concentrations. In contrast, the risk of abnormality was not significantly higher as toxic equivalent concentrations increased in eggs. We also found significant 7-ethoxyresorufin O-deethylase and Cytochrome P4501A responses in livers of hatchling turtles from Lake Ontario relative to hatchlings from a clean, inland site whereas we did not find any evidence of porphyria in the hatchlings from either site.

Immunoquantitation of cytochromes P450 1A and P450 2B and comparison with chlorinated hydrocarbon levels in archived polar bear liver samples.

The present study examined the utility of an immunoblot method for quantitation of cytochrome P450 isozymes in archived liver samples as a bioassay of exposure to halogenated hydrocarbons. Hepatic microsomes were prepared from 44 archived polar bear (Ursus maritimus) liver homogenates that had been stored at approximately -40 degrees C for 9-10 years and analyzed on blots probed with antibodies to rat cytochromes P450 1A1 and P450 2B1. The results revealed a positive correlation between cytochrome P450 1A and total polychlorinated biphenyl (PCB) levels in the archived liver samples, suggesting that cytochrome P450 1A was induced in polar bears by environmental exposure to PCBs.