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1.
Newark Bay (NB) killifish (Fundulus heteroclitus) have been chronically exposed to environmental contaminants that activate the aryl hydrocarbon receptor (AHR) and are tolerant to toxic effects and CYP1A induction provoked by AHR ligands. Resistance to CYP1A induction could be due to an epigenetic mechanism such as DNA methylation. We measured in-ovo CYP1A catalytic activity (ethoxyresorufin-O-deethylase, EROD) in NB and reference site killifish embryos aqueously exposed to various concentrations of the de-methylating agent 5-azacytidine, 5-AC (5, 50 and 500 micro(micro)M) with or without 0.2 micro(micro)g/l of the CYP1A inducer 3,3',4,4',5 pentachlorobiphenyl (IUPAC PCB126). Neither PCB126 alone, nor PCB126 plus 5-AC, induced EROD above levels in vehicle treated Newark Bay fish. In reference site fish, the same PCB126 dose provoked a 7.4-fold EROD induction relative to controls. We conclude that Newark Bay killifish are resistant to CYP1A induction by co-planar PCBs during early embryological development and our data suggests that DNA methylation does not play a critical role in resistance to CYP1A induction in this model.  相似文献   

2.
Newark Bay (NB) killifish (Fundulus heteroclitus) have been chronically exposed to environmental contaminants that activate the aryl hydrocarbon receptor (AHR) and are tolerant to toxic effects and CYP1A induction provoked by AHR ligands. Resistance to CYP1A induction could be due to an epigenetic mechanism such as DNA methylation. We measured in-ovo CYP1A catalytic activity (ethoxyresorufin-O-deethylase, EROD) in NB and reference site killifish embryos aqueously exposed to various concentrations of the de-methylating agent 5-azacytidine, 5-AC (5, 50 and 500 μ(micro)M) with or without 0.2 μ(micro)g/l of the CYP1A inducer 3,3,4,4,5 pentachlorobiphenyl (IUPAC PCB126). Neither PCB126 alone, nor PCB126 plus 5-AC, induced EROD above levels in vehicle treated Newark Bay fish. In reference site fish, the same PCB126 dose provoked a 7.4-fold EROD induction relative to controls. We conclude that Newark Bay killifish are resistant to CYP1A induction by co-planar PCBs during early embryological development and our data suggests that DNA methylation does not play a critical role in resistance to CYP1A induction in this model.  相似文献   

3.
CYP1A-immunopositive protein can be elevated in response to planar PAHs and PCBs in Mytilus sp. digestive gland whilst CYP3A-immunopositive protein has been associated with testosterone 6beta-hydroxylation in fish. Levels of CYP1A- and CYP3A-immunopositive protein were determined in Mytilus galloprovincialis digestive gland microsomes collected from 12 sites in the Mediterranean Sea during May and September 2001. CYP1A-immunopositive protein was significantly highest at contaminated sites whilst CYP3A-immunopositive protein was significantly lowest. A weak negative correlation (r2 = 0.21) was seen between CYP1A- and CYP3A-immunopositive protein. Little evidence of differences at the different sampling times was observed. These results confirm previous work indicating elevation of CYP1A-immunopositive protein in Mytilus sp. digestive gland at contaminated sites. Further study is required to characterise CYP3A-like expression in Mytilus and to elucidate the consequences of possible CYP3A-like down-regulation at contaminated sites.  相似文献   

4.
CYP1A-immunopositive protein can be elevated in response to planar PAHs and PCBs in Mytilus sp. digestive gland whilst CYP3A-immunopositive protein has been associated with testosterone 6β-hydroxylation in fish. Levels of CYP1A- and CYP3A-immunopositive protein were determined in Mytilus galloprovincialis digestive gland microsomes collected from 12 sites in the Mediterranean Sea during May and September 2001. CYP1A-immunopositive protein was significantly highest at contaminated sites whilst CYP3A-immunopositive protein was significantly lowest. A weak negative correlation (r2=0.21) was seen between CYP1A- and CYP3A-immunopositive protein. Little evidence of differences at the different sampling times was observed. These results confirm previous work indicating elevation of CYP1A-immunopositive protein in Mytilus sp. digestive gland at contaminated sites. Further study is required to characterise CYP3A-like expression in Mytilus and to elucidate the consequences of possible CYP3A-like down-regulation at contaminated sites.  相似文献   

5.
Halogenated aromatic hydrocarbons (HAHs) and polynuclear aromatic hydrocarbons (PAHs) are major environmental contaminants. Fish species that are chronically exposed to these compounds can develop resistance to their toxic effects. In all fish species studied to date, toxicant resistance has been accompanied by decreased inducibility of the xenobiotic metabolizing enzyme, cytochrome P450 1A (CYPIA). CYP1A induction is mediated through the Aryl Hydrocarbon Receptor (AHR). Although these compounds mediate their effects through this pathway, there have been resistant populations in which one chemical class cannot induce CYPIA expression (HAHs) while the other (PAHs) can. Resistance to PAHs was examined in a HAH-resistant population of Fundulus heteroclitus collected from a site contaminated with both compound classes (Newark Bay, NJ). Fish were injected intraperitoneally with the HAH 3,4,3',4'-tetrachlorobiphenyl (PCB77), benzo[a]pyrene (B[a]P, a PAH) or vehicle and sacrificed after 2 (B[a]P) or 5 days (PCB77, vehicle). We found no significant increase in CYP1A mRNA levels in resistant Newark Bay F. heteroclitus treated with either B[a]P or PCB77, while there was a 3.9 fold (PCB77) and 4.2 fold (B[a]P) increase in CYP1A mRNA in Flax fish relative to controls. AHR labeling studies revealed significantly (P < 0.05) lower levels of hepatic AHR in Newark fish (1,770 +/- 1,693.2 DPM) relative to Flax fish (6,082.5 +/- 1,709.9 DPM). Overall, these data suggest Newark F. heteroclitus are resistant to both PAHs and HAHs at the level of CYP1A mRNA, which might be mediated, in part, though lower expression of AHR. We are currently studying the promoter sequence to determine its role in chemical resistance.  相似文献   

6.
Mummichogs, Fundulus heteroclitus, an estuarine fish with a relatively small home range found along the eastern coast of the United States are well-suited to monitoring contaminant effects, including those of polycyclic aromatic hydrocarbons (PAHs). One of the common PAHs in estuaries is pyrene. We report here on efforts to develop multiple biomarkers of pyrene exposure in this species. Adult male mummichogs were exposed in the laboratory to the weak aryl hydrocarbon receptor (AhR) agonist pyrene at 0, 30, or 50 microg/L in 7-day static renewal exposures. The RNA was extracted from livers and alterations in mRNA expression were assessed by subtractive hybridization and differential display in order to produce multiple biomarkers of pyrene exposure. Genes demonstrating differential expression were confirmed by quantitative-PCR (Q-PCR) and include cytochrome P-450 1A (CYP1A), a putative hepatocyte growth factor activator, a X-ray inducible retrotransposon, and several expressed sequenced tags (ESTs). Some of these genes represent new biomarkers of pyrene exposure and potential biomarkers of PAH exposure. Therefore, similar changes were investigated at a Superfund site in Charleston, SC. Mummichogs from a creosote contaminated site and from a reference site (North Inlet National Estuarine Research Reserve near Georgetown, SC) were trapped, RNA extracted from the livers, and Q-PCR performed. Many of the genes differentially expressed following pyrene exposure were not altered at the creosote contaminated site in comparison to the reference site. However, CYP1A and an EST were induced. CYP1A induction at Diesel Creek indicates that this population of fish does not demonstrate refractory CYP1A phenotypes observed at several sites with high levels of AhR agonists. Ultimately, we anticipate that the use of multiple biomarkers of PAH exposure will provide useful information on the potential effects of toxicants.  相似文献   

7.
Induction of hepatic cytochrome P450-dependent microsomal mono-oxygenase by xenobiotics is a well-established phenomenon in teleost fish. As in laboratory mammals, fish possess multiple forms of cytochrome P450 that display overlapping substrate specificity. One such isoform, CYP1A1, which has been cloned and sequenced from rainbow trout, has been shown to be orthologous to rat CYP1A1 and, as in mammals, is inducible up to several hundred-fold by planar aromatic hydrocarbons, PCBs and dioxins. It has been suggested that induction of CYP1A1 orthologues might provide a sensitive biomonitor for environmental pollution by mixtures of such compounds. In the current study, polyclonal antibodies directed against CYP1A1 purified from rat and trout liver were used to monitor induction of the CYP1A1 orthologue in hepatic microsomes from the fresh water species, the channel catfish (Ictalurus punctatus). Catfish from a local fish farm were induced in the laboratory by three daily injections of 50 mg/kg of the PCB mixture Aroclor 1254 and compared with fish taken from a site in central Arkansas—the Bayou Meto, known to be polluted with dioxin. Hepatic microsomal activities towards ethoxyresorufin (EROD) and pentoxyresorufin (PROD) were measured and Western blot analysis carried out with the two antibodies. EROD was elevated in both the Aroclor-treated fish and in the Bayou Meto fish compared with untreated fish farm controls; smaller but significant increases were observed in PROD. Spearman's rank correlations of 0·74 and 0·89 were observed between EROD and immunoquantified cross-reactivity towards the rat CYP1A1 and trout CYP1A1 antibodies.  相似文献   

8.
9.
Concentrations of PCBs and their toxic potential were examined in subcutaneous fat of eight albatross and one petrel species collected from the North Pacific and the Southern Oceans. Among all the species analyzed, high PCB levels were found in adult male blackfooted albatross from the North Pacific with the mean value of 92 microg/g wet weight. No significant gender difference in PCB accumulation was observed (P>0.1). The mean PCB levels in Southern Oceanic birds were 1 or 2 orders of magnitude lower than those from the North Pacific albatrosses. A regional-specific accumulation of non-ortho coplanar congeners were observed, most birds from the Southern Ocean had higher IUPAC 169 levels while IUPAC 126 concentrations were higher in those from the North Pacific. The estimated toxic equivalents for black-footed and Laysan albatrosses from the North Pacific were in the same range of some fish-eating birds, which were highly contaminated by PCBs. The correlation between ratio of IUPAC 169/126 concentration and total PCBs concentration indicated the possibility of induction in cytochrome P450 activities in North Pacific albatrosses (P<0.01). The calculated hazard indices indicated that black-footed and Laysan albatrosses inhabiting in the North Pacific had similar threshold levels which were known to cause toxic effects in some populations of fish-eating birds.  相似文献   

10.
Mytilus edulis were collected from a reference site (Port Quin) and an urban/industrial contaminated site (New Brighton) in the UK during June 1999. Levels of PCBs (sigma7 congeners) and CB-138 were determined to be, respectively, 21 fold and 16 fold higher in the mussel digestive glands from New Brighton. Levels of CYPIA-immunopositive protein were 1.5 fold higher (P < 0.05) at the polluted site but the levels of DNA strand breaks were 1.3 fold higher (P<0.05) at the reference site. Mussels from Port Quin were placed in cages at both sites and both transplanted and indigenous populations sampled in September (13 weeks). Mussels transplanted from the reference site to the industrial site, reported elevated levels of CYP1A-immunopositive protein (1.4 fold; P < 0.05) and higher levels of DNA damage (1.2 fold; P < 0.05) compared to caged populations at the reference site and a PCB loading similar to the populations from the polluted site. Moreover, transplanted mussels had DNA damage 1.8 fold greater (P < 0.05) than indigenous mussels at the transplant site. These changes were small but significant when compared to the observed temporal changes in the indigenous populations.  相似文献   

11.
To assess chemical contaminant stress in the marine environment, ethoxyresorufin-O-deethylase (EROD) activity and cytochrome P450 1A (CYP1A) expression were measured in 88 English Sole (Pleuronectes vetulus) collected during May and June 1999 from four sites in Vancouver Harbour and at an expected reference site outside the harbour. Hepatic microsomes were prepared from the fish and analyzed for total CYP content, EROD activity, and CYP1A protein levels. Hepatic EROD activity and CYP1A protein levels were elevated in fish from two sites in the inner harbour. A comparison with sediment chemistry data showed that fish with increased EROD activity and CYP1A levels came from sites containing relatively high levels of polycyclic aromatic hydrocarbons and polychlorinated biphenyls. Unexpectedly high levels of EROD activity and CYP1A protein were also found in fish from a reference site near Gibsons, in Howe Sound. The elevated EROD activity and CYP1A expression in fish from this site cannot be explained by the chemical analysis data collected.  相似文献   

12.
Experimental exposure tanks were set up containing 'clean' marine sediment spiked with set doses of polyaromatic hydrocarbons (PAH), or polychlorinated biphenyls (PCB), or both. Reference tanks contained only 'clean' sediment. Female dab Limanda limanda L., exposed to these sediments for 7 days showed no differences in serum total protein concentration, serum lysozyme activity or body growth. Their kidney leucocytes were assayed for extracellular production of reactive oxygen species of the phagocytic respiratory burst. No significant differences in superoxide anion (O2-) production were found between fish exposed to the 'spiked' sediments and those on the 'clean' reference sediment. The production of hydrogen peroxide (H2O2) was, however, reduced in the fish held on the PAH and PAH/PCB spiked sediments. H2O2 levels for fish exposed to PCBs alone, did not differ significantly from those of control fish. Our results suggest that the decrease in H2O2 production was due to exposure to PAHs, rather than to PCBs.  相似文献   

13.
Mytilus edulis digestive gland microsomes were prepared from indigenous populations sampled from a clean reference site (Port Quin) and an urban-industrial contaminated site (Blackpool) in the UK. Samples were collected in March/April, May, August and December 1998. Western blot analysis was performed using polyclonal antibodies to fish CYP1A and rat CYP2E using partially purified M. edulis CYP as a positive control, to aid identification. CYP1A- and CYP2E-immunopositive protein levels showed different site-specific seasonal variation with higher levels of CYP2E determined in May (P < 0.05). At both sites, lower levels of CYP1A-immunopositive protein but not CYP2E-immunopositive protein were observed in the samples collected in December (P < 0.05). This correlated with lower levels of nuclear DNA damage (Comet assay expressed as per cent tail DNA) observed in December compared to August (P < 0.05).  相似文献   

14.
Reproductively active oysters were fed daily with 0.2 g algal paste containing 0, 0.1, and 1.0 microgram polychlorinated biphenyls (PCBs) (1:1:1 mixture of Aroclor 1242, 1254 and 1260) for either 15 or 30 days, and accumulation of PCBs in different organ tissues and eggs assessed. The effects of PCB exposure on lipid content, lipid class and fatty acid composition were also evaluated. PCBs were accumulated by the oysters and transferred to the eggs. PCB accumulation in oysters was dose, time and tissue dependent. Mean PCB contents were 3150, 1970, and 250 ng/g dry wt., respectively, in the visceral mass, gills + mantle and muscle of oysters fed algal paste containing 1.0 microgram PCBs for 30 days. The PCBs in the eggs from the same oysters reached 671 ng PCBs/g dry wt. Feeding oysters with PCB-sorbed algal paste for 30 days significantly increased phospholipid and free fatty acid contents in gills + mantle tissue compared to the same tissues in the undosed control.  相似文献   

15.
Reproductively active oysters were fed daily with 0.2 g algal paste containing 0, 0.1, and 1.0 μg polychlorinated biphenyls (PCBs) (1:1:1 mixture of Aroclor 1242, 1254 and 1260) for either 15 or 30 days, and accumulation of PCBs in different organ tissues and eggs assessed. The effects of PCB exposure on lipid content, lipid class and fatty acid composition were also evaluated. PCBs were accumulated by the oysters and transferred to the eggs. PCB accumulation in oysters was dose, time and tissue dependent. Mean PCB contents were 3150, 1970, and 250 ng/g dry wt., respectively, in the visceral mass, gills+mantle and muscle of oysters fed algal paste containing 1.0 μg PCBs for 30 days. The PCBs in the eggs from the same oysters reached 671 ng PCBs/g dry wt. Feeding oysters with PCB-sorbed algal paste for 30 days significantly increased phospholipid and free fatty acid contents in gills+mantle tissues compared to the same tissues in the undosed control.  相似文献   

16.
Selective accumulation of PCDDs, PCDFs and PCB congeners was investigated in beluga (Delphinapterus leucas), narwhal (Monodon monoceros) and several species at the same trophic level in two areas of Canada. There was a much higher relative abundance of meta-para-unsubstituted PCBs in the cetaceans compared with other species. This suggests that activity of cytochrome P450 monooxygenase enzymes causing CYP2B-type metabolism was relatively low in beluga and narwhal. Lower relative levels of 2,3,7,8-TCDD occurred in beluga from both areas, and selective reduction of toxic non-ortho PCBs occurred in the highly PCB-contaminated St Lawrence beluga. These compounds are potent inducers of cytochrome P450 CYP 1A-type enzymes, suggesting that such an enzyme with the capability of metabolizing TCDD-like substrates is present in beluga and narwhal. SIMCA principal components analysis of PCB congener patterns showed that the two cetaceans were in a statistically significant different class than the other species. Variables (congeners) with the greatest separation power between classes were mainly those defined by the selective metabolism rules.  相似文献   

17.
Killifish (Fundulus heteroclitus) from a highly contaminated site on the Elizabeth River are resistant to the acute toxicity and the cytochrome P4501A (CYP1A)-inducing activity of both the sediments from the site and chemically pure polycyclic aromatic hydrocarbons (PAHs). These effects are highly heritable for one generation, but heritable to a lesser degree by subsequent generations, in clean conditions in the laboratory. We show that offspring of this population of Elizabeth River killifish are also resistant to the teratogenicity and P4501A-inducing activity of PCB congener 126, a prototypical coplanar halogenated aromatic hydrocarbon (HAH). Furthermore, the pattern of greater resistance to acute toxicity and P4501A-inducing activity in the first generation and less in subsequent generations is also observed upon exposure to PCB-126.  相似文献   

18.
19.
A laboratory experiment was conducted to investigate the transfer of organic contaminants from an environmentally contaminated marine sediment through a simple marine food chain. The infaunal polychaete, Nereis virens, was exposed to contaminated sediment collected from the Passaic River, NJ, USA, for 70 days. These polychaetes were then fed to the American lobster, Homarus americanus, for up to 112 days. Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), 2,4,6,8-tetrachlorodibenzothiophene (TCDT), polychlorinated biphenyls (PCBs), and several chlorinated pesticides were accumulated by polychaetes following exposure to the contaminated sediment. Some of these contaminants were also accumulated by lobsters which were exposed to the contaminated sediment and/or fed contaminated polychaetes. Only the lesser chlorinated PCDDs and PCDFs (mostly tetra- and pentachlorinated congeners) and 2,4,6,8-TCDT were detected in the polychaetes and lobster. Significant alterations were noted in the PCB patterns found in both species, particularly the lobster. The non-ortho-substituted PCBs (such as congeners 77 and 126) became enriched in the PCB mixtures of the polychaetes and especially the lobsters relative to the sediment, probably because these congeners were not metabolized. These congeners and the 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity equivalents of the PCB mixtures were enriched by a factor of about six in the lobsters relative to the sediment. Elimination of PCB congeners containing vicinal hydrogens in the meta-para region is consistent with cytochrome P450IIB-type metabolism. Based on the concentration trends for some PCB congeners and chlorinated pesticide ratios measured in the lobsters during this experiment, it appears that this metabolic system is inducible in the American lobster.  相似文献   

20.
Sediments and organisms were examined for concentrations of organic and metal contaminants from near the Los Angeles County (JWPCP) municipal outfall at Palos Verdes (PV) station 7-3, the Los Angeles City (Hyperion) municipal outfall at Santa Monica Bay (SMB) station 6-4 and reference station SMB 2–3 near Malibu Beach. Flows and mass emission rates of suspended solids, PCBs, Cd and Zn were similar at the two outfalls. Mass emission rate of copper was almost twice as high from Hyperion as from JWPCP, while mass emission rate of DDTs was an order or magnitude higher from JWPCP than from Hyperion.Surficial sediments at PV 7-3 were enriched in most contaminants relative to SMB 6-4 and relative to the mass emission rates of contaminants from the JGVPCP and Hyperion outfalls. Some of this enrichment could be accounted for by the greater accumulation of organic material, measured as total volatile solids, at PV 7-3 relative to SMB 6-4. Some might be accounted for by resurfacing of more contaminated historical deposits buried at PV 7-3. Some of the enrichment of DDTs relative to PCBs could be accounted for by the greater abundance of oxygenated metabolites of PCBs (PCBols) relative to DDTs (DDTols) in sediments.The degree of contamination of organisms by DDTs increased with proximity to PV 7-3 but contamination by PCBs was similar at PV 7-3 and SMB 6-4. DDT concentrations in fish livers ranged from 12 ± 4 ( ) mg/wet kg in longspine combfish from SMB 2–3 to 610 ± 105 (n = 5) mg/wet kg in Pacific sanddab from PV 7-3. DDT concentrations in fish gonads ranged from 0·003 ± 0·003 (n = 5) mg/wet kg in yellowchin sculpin from SMB 6-4 to 1.5 ± 6 (n = 3) mg/wet kg in Pacific sanddabs from PV 7-3. PCB concentrations in fish livers ranged from 1·2 ± 0·4 (n = 4) mg/wet kg in yellowchin sculpin from SMB 2–3 to 16 ± 3 (n = 6) in Pacific Sanddab from SMB 6-4. DDT and PCB concentrations in invertebrate hepatopancreas were only slightly lower than those in fish livers. DDTols and PCBoIs comprised an average of 91 % of the total of parent compounds and oxygenated metabolites in sediments and 66 % in livers and hepatopancreas. Trace metals were frequently decreased in livers and hepatopancreas from near outfalls even though they were highly elevated in sediments.Comparison of sediment and tissue chlorinated hydrocarbon data with that from Elliot and Commencement Bays, Puget Sound, indicated that none of the southern California coastal stations considered in this study were sufficiently lacking contamination to be considered as adequate control sites.  相似文献   

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