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Robert B. Spies David W. Rice Jr Paul A. Montagnaand Robert R. Ireland 《Marine environmental research》1985,17(2-4)
To identify some specific effects of organic contaminants on fisheries in an urbanized estuary we compared the reproductive success of starry flounder from San Francisco Bay with concentrations of tissue contaminants and hepatic mixed-function oxidase (MFO) activity. We found significantly lower (P < 0·05) sediment concentrations of total identified polynuclear aromatic hydrocarbons (PAHs) in the less urbanized San Pablo Bay (SP) area (Fig. 1) than in the more urbanized central bay (CB) stations (Table 1). For flounder in early gametogenesis (August and September) the SP fish (n = 20) had significantly lower (P < 0·01) liver concentrations of Aroclor 1260 (0·34 ± 0·14 μg/g) than those at the CB stations: Berkeley (BK, n = 20, 1·6 ± 1·6 μg/g); Oakland (OK, n = 16, 2·3 ± 2·8 μg/g); and Alameda (AL, n = 4, 2·2 ± 1 μg/g). A similar pattern existed for DDT concentrations: SP = 0·2 ± 0·16 μg/g; BK = 0·1 ± 0·34 μg/g; OK = 0·4 ± 0·53 μg/g; and AL = 0·4 ± 0·33 μg/g. Total PAHs in livers were as follows: SP = 0·14 μg/g; BK = 2·6 μg/g; OK = 1·4 μg/g; and AL = 14 μg/g. Although gonad index, liver index, and presence of fin rot are inversely related to aryl hydrocarbon hydroxylase (AHH) activity, healthy fish in a similar reproductive state have lower AHH activities in the SP area. For example, in August and September, 1984, mean AHH activities were as follows: SP = 203 ± 89, and CB = 355 ± 200 pmol 3-OH-B[a]P mg microsomal protein min. We found a log-linear relationship for AHH activity and its percent inhibition by 7,8-benzoflavone (10−4m) and only a few fish from SP showed enhanced AHH activity after addition of 7,8-benzoflavone. This suggests that most of the starry flounder in San Francisco Bay are induced. 相似文献
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Hepatic CYP1A levels and EROD activity in English sole: biomonitoring of marine contaminants in Vancouver Harbour 总被引：4，自引：0，他引：4
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
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Juveniles of both stone flounder Platichthys bicoloratus and starry flounder Platichthys stellatus utilize estuaries as nursery grounds. To understand their habitat selection and the functions of habitats such as food supply, we defined the seasonal distribution of recently settled fish of these species in shallow nursery areas and investigated their feeding habits in the Natori River estuary, Japan. Distribution of stone flounder was limited to the lower estuary (<3 km upriver from the mouth) and stone flounder were most abundant near the mouth. Recently settled starry flounder were first detected further upstream in areas characterized by low salinity <10 and by the absence of the predatory sand shrimp Crangon uritai. Early juvenile stone and starry flounders consumed mainly siphons of the bivalve Nuttallia olivacea and the mysid Neomysis awatschensis, respectively; however, 1- and 2-yr-old fish of both stone and starry flounders fed mainly on the bivalve siphons. These results indicate that habitat selections of juvenile stone and starry flounders enable utilization of preferred prey and predator avoidance, respectively, and that non-overlap of these species' habitats results in avoidance of inter-specific competition for food between these two species. 相似文献
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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
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Nicholas H. Vrolijk Nancy M. Targett Bruce R. Woodin John J. Stegeman 《Marine environmental research》1995,39(1-4)
The relationship between cytochrome P450 and feeding on terpenoid-rich gorgonian corals was investigated in a species of tropical butterflyfish and compared with two other sympatric congeners that do not feed on gorgonians. Fish were collected from non-polluted waters in Belize and the levels of two cytochrome P450 isozymes (CYP2B and CYP3A) were immunoquantitated in addition to quantification of total P450. Chaetodon capistratus regularly feeds on gorgonian corals and has higher levels of total hepatic microsomal cytochrome P450 than C. ocellatus or C. striatus. The content of hepatic P450 (0.588–0.794 nmol mg−1) in C. capistratus is among the highest ever reported in teleosts from non-polluted waters and is significantly greater than detected in C. ocellatus or C. striatus. Chaetodon capistratus also had a larger hepatic index (g liver per g fish) and more microsomal protein (mg protein per g liver), factors that translate into 3.3- to 8-fold more total P450 per g fish. Sexual differences in total P450 were observed between male and female C. capistratus, but not among the other species. The contents of proteins detected by immunoassay with polyclonal anti-scup P450B (CYP2B) and anti-human P4503A (CYP3A) were 2- to 10-fold and 2- to 20-fold greater, respectively, in C. capistratus than in the congeneric species. CYP2 and CYP3 gene families in mammals are thought to have evolved partially in response to dietary allelochemicals. These results suggest that these P450 isozymes may also be important in marine teleosts that feed on terpenoid-rich prey. 相似文献
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From sediment bioassay to fish biomarker--connecting the dots using simple trophic relationships. 总被引：1，自引：0，他引：1
C A Rice M S Myers M L Willis B L French E Casillas 《Marine environmental research》2000,50(1-5):527-533
Two common problems in applying and interpreting invertebrate bioassays and fish biomarkers in sediment toxicology are the wide gap between significant effects concentrations determined by these two approaches, and a general lack of ecological context. We have devised an exposure system that is able to reconcile much of the disparity between invertebrate bioassay and fish biomarker results by incorporating realistic ecological processes based on deposit feeding and predator-prey interactions. This system relates the disturbance of interest (sediment contamination) to biologically meaningful effects in a resource of interest (marine flatfish) via a realistic contaminant vector (a deposit-feeding polychaete worm). In this pilot study, polychaetes (Armandia brevis) were exposed for 28 days to clean sediments supplemented with benzo(a)pyrene (BaP), para-para dichlorodiphenyldichloroethylene (pp'DDE), Aroclor 1254, or field sediments collected from two sites in Puget Sound, Washington, contaminated predominantly with polcyclic aromatic hydrocarbons (PAHs) or chlorinated compounds. Exposed worms were then fed live to juvenile English sole (Pleuronectes vetulus) for 10 or 12 days. At the end of the exposure period, fish were measured for length and weight, sacrificed, and preserved for either routine histopathology and immunohistochemical analysis of cytochrome P450 1A induction, or 32P post-labeling determination of hepatic PAH-DNA adducts. Growth of predatory flatfish was lower than reference in all but one of eight groups fed contaminant-exposed polychaetes; however, statistically significant reductions in growth were only observed in three of these eight groups, at least in part due to low statistical power. Juvenile sole from all contaminant-exposed groups showed increased expression of CYP1A, and fish exposed to BaP-exposed worms showed clear evidence of hepatic PAH-DNA adducts. This method allows the concurrent evaluation of sediment contamination at multiple biological and ecological levels. These results indicate that sediments determined to be nontoxic by common invertebrate bioassays may have the potential to cause adverse effects at higher trophic levels. 相似文献
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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
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C A Godard M J Leaver M R Said R L Dickerson S George J J Stegeman 《Marine environmental research》2000,50(1-5):7-10
The cytochromes P450 (CYP) constitute a multigene family of enzymes playing a critical role in the oxidation of many endogenous and xenobiotic substrates. The CYP1 family is of particular interest in environmental toxicology because its members are dominant in the metabolism of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and aryl amines. Three members of the CYP1 family, CYP1A1, CYP1A2, and CYP1B1, have been identified in mammals. We report here on the identification and cloning of cytochrome P4501B-like sequences from two teleost fish species and a marine mammal. Sequences clustering with CYP1B1 in phylogenetic analysis were obtained from liver cDNA of scup (Stenotomus chrysops), genomic DNA of plaice (Pleuronectes platessa), and liver cDNA of striped dolphin (Stenella coeruleoalba). 相似文献
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A. Khler B. Lauritzen S. Banns S. G. George L. Frlin C. J. F. Van Noorden 《Marine environmental research》1998,46(1-5)
Progression from eosinophilic foci to persistent basophilic foci and carcinomas was observed in pollution-induced hepatocellular carcinogenesis in European flounder (Platichthys flesus L.) in a similar sequence as in chemically induced liver cancer in mammals. Image analysis was used to quantify enzyme activity and protein expression as visualised by enzyme and immunohistochemistry in various stages of toxipathic lesions and carcinogenesis. In eosinophilic foci, growth advantage was achieved by increased production of NADPH and pentoses for biosynthesis and cell proliferation due to increased G6PDH capacity during the first steps of clonal adaptation. Simultaneously, oxyradical production by CYP450, phase I was reduced. Overexpression of P-gp mediating multi-xenobiotic resistance was noted in basophilic cell types which persisted during progression towards carcinomas. This was accompanied by increased protein levels of oxyradical scavenging GST-A. These changes are consistent with adaptation and phenotypic expression of the multidrug or xenobiotic resistance (MDR/MXR) type in hepatocarcinomas of European flounder. 相似文献
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The contaminant-induced cytochrome P450 (CYP) members in minke whale (Balaenoptera acutorostrata) can be potential biomarkers of the contaminant exposure and toxic effects. In this study, we constructed a cDNA library from the liver of minke whale from the North Pacific, and further screened a total of 6930 clones randomly selected in the library for the isolation of cDNA clones encoding novel members of CYP superfamily. The screening revealed the isolation of six novel CYP cDNA clones that are classified into CYP1A, CYP2C, CYP2E, CYP3A, CYP4, and CYP4A subfamilies. The BLAST homology search using the partial cDNA fragments of four CYP subfamilies (CYP1A, CYP2C, CYP2E and CYP4A) demonstrated that the minke whale CYPs were most closely related to pig CYPs (81-91%). Identification of multiple CYP genes in marine mammal species such as minke whale will provide new insights into the metabolic or toxicological functions of individual CYP members. 相似文献
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Michael R. Miller David E. Hinton James J. Blair John J. Stegeman 《Marine environmental research》1988,24(1-4)
Monoclonal antibody directed against a major β-naphthoflavone (BNF)-induced form of teleost cytochrome P-450, P-450E (equivalent to P-450c in rat) was used to immunolocalize this enzyme in liver, gill and heart of scup and trout. Liver sections from both species showed P-450E in the cytoplasm of hepatocytes. No regional differences were observed which might indicate zonation of cytochrome P-450E within subpopulations of hepatocytes. Scup exocrine pancreatic cells were only weakly positive. In the gill of both fish, cytochrome P-450E was restricted to the endothelium (pillar cells) of secondary lamellae, where fluorescence appeared as a chain in longitudinal sections through lamellae and as star-shaped clusters in en face views. Sections of ventricular wall in both species revealed P-450E was restricted to endothelium at margins of muscle bands limiting heart ventricular lumen. Localization in the specific cells of these and other organs may be fundamentally important in understanding the role of cytochrome P-450E. 相似文献
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J. W. Anderson J. M. Jones S. Steinert B. Sanders J. Means D. McMillin T. Vu R. Tukey 《Marine environmental research》1999,48(4-5)
In 1993 collections of marine mussels (Mytilus galloprovincialis) were deployed 1 m from the water surface at six sites in San Diego Bay for 88 days. A similar mussel deployment was conducted in 1995, except the animals were deployed 1 m off the bottom and only for 32 days. After recovery from the sites, tissue was extracted with dichloromethane and the solvent extracts analyzed for chemical contaminant content and the ability to produce CYP1A1 induction in a transgenic cell line (TV101L cells). The cells used in the assay (P450 RGS) are stably transfected with a plasmid containing firefly luciferase linked to human CYP1A1 promoter sequences. Induction (fold increase compared to control) was determined by luminometry 16 h after application of small volumes (2–10 μl) of solvent extracts to cultured cells. Small mussels deployed in the Naval Station (NAV) in 1993 exhibited very high bioaccumulation of polycyclic aromatic hydrocarbons (PAHs; 52 μg/g) and polychlorinated biphenyls (PCBs), in addition to very strong induction of CYP1A1 measured by reporter gene system (RGS) responses. Large mussels deployed at the NAV station in 1993 and intermediate-sized animals placed at three stations within the NAV station in 1995 accumulated 13–29 μg PAH/g and exhibited relatively high RGS responses. Correlation of RGS responses for all mussel samples to the measured PAH concentrations was 0.85 (r2). When the concentrations of seven specific PAHs found in the samples are converted to benzo[a]pyrene equivalents, from previously derived toxic equivalency factors (TEFs) for this test system, and compared to measured RGS responses, the correlations are approximately 0.9. The results of these studies indicate that the RGS biomarker can be used as a screening tool for detection of CYP1A1-inducing compounds in tissues, and an estimate of potential human health or ecological risk from ingestion of contaminated organisms. Positive RGS responses can be followed by detailed chemical analyses of PAHs and coplanar PCBs using the same extract. 相似文献
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The major glutathione S-transferase isoform of flounder liver, an antigenically related structural homologue of plaice GST-A, also displays mRNA homology. A 901bp cRNA probe for plaice GST-A cross-hybridised to a 1100bp flounder mRNA on northern blot analysis. The plaice antibody and cRNA probes were used to study effects of inducer treatment on GST-A expression in flounder liver. Six days after PAH treatment (3-methylcholanthrene) total hepatic GST activity was halved, levels of GST-A were 80% and GST-A mRNA levels were 25% of controls. A commercial PCB mixture (Aroclor 1254TM) had little effect on total GST or GST-A levels despite halving GST-A mRNA levels. An epoxide, trans-stilbene oxide induced total GST activity 1·4 fold and GST-A protein levels 1·8-fold and its mRNA levels 3-fold. This reduced expression of the major flounder hepatic GST by agents which induce cytochrome P4501A1 may modulate cytoxicity of environmental pollutants in this species. 相似文献
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Winter flounder (Pleuronectes americanus) from Boston Harbor, Massachusett, undergo a progressive series of hepatic changes involving hydropic vacuolation of epithelial cells, biliary hyperplasia, and hepatocellular and cholangiocellular neoplasia. Severely affected fish often exhibit grossly visible lesions. To examine cell proliferation associated with these conditions, evidence for DNA synthesis was sought. Boston winter flounder livers were screened endoscopically to select fish with and without grossly visible hepatic lesions, and then injected intraperitoneally with bromodeoxyuridine (BrdU) 3 h before euthanasia. Incorporation of this nucleotide analog was visualized immunohistochemically in formalin-fixed, paraffin-embedded tissues. Basal epithelia in the gill and intestine stained strongly, as did renal hemopoietic epithelia. In contrast, renal tubular epithelia and cardiomyocytes did not stain. Normal appearing hepatocytes stained occasionally, whilst more frequent staining was observed in vacuolated and neoplastic hepatic epithelia. We conclude (1) that the BrdU assay as described here has potentially widespread application to the study of cell proliferation in fish tissues; (2) that vacuolated cells may actively proliferate in winter flounder liver from Boston Harbor; and (3) vacuolated cells may be involved in the neoplastic transformation of hepatic epithelia. 相似文献
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G. D. James 《新西兰海洋与淡水研究杂志》2013,47(3):229-240
Cod‐end mesh selection experiments were conducted on three species of flatfish (common or English sole Peltorhamphus novaezeelandiae, lemon sole Pelotretis flavilatus, and sand flounder Rhombosolea plebeia) trawled off the Otago coast. It is suggested that better yields would be obtained from stocks of these fish if the minimum cod‐end mesh size was increased from 10.2 cm (4 in.) to 11.4 cm (4.5 in.). 相似文献