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1.
The uptake of polycyclic aromatic hydrocarbons (PAHs) by marine deposit-feeding invertebrates can be determined by screening for PAH-derived metabolites. We identified 1-hydroxypyrene as the only intermediate metabolite in tissue of four species of deposit-feeding polychaetes, Nereis diversicolor, Nereis virens, Arenicola marina, and Capitella sp. I exposed to pyrene spiked sediment. Synchronous fluorescence spectroscopy (SFS) provides a fast and simple method for both qualitative and quantitative analysis of 1-hydroxypyrene in all four species. The SFS assay was validated using HPLC with ultraviolet detection. A good correlation between 1-hydroxypyrene concentrations determined by the two methods was observed. We used HPLC with fluorescence detection combined with enzymatic hydrolysis of conjugated metabolites to investigate species specific metabolite patterns. A tentative aqueous metabolite identification scheme indicates that Nereid polychaetes predominately make use of glucuronide conjugation whereas Capitella sp. I. and Arenicola marina appear to utilize predominantly sulfate and/or glucoside conjugation. The usefulness of 1-hydroxypyrene as a biomarker for PAH exposure in deposit-feeding invertebrates is discussed. 相似文献
2.
Pollutants such polycyclic aromatic hydrocarbons (PAHs) are released into the environment by urban communities and industries and the enzymes that catalyse the biotransformation of pollutants play a key role regarding the accumulation of these compounds in fish species inhabiting these areas. In this study the relationship between phase I (EROD activity) and phase II (GST activity) and PAH metabolites was measured in grey mullet (Mugil cephalus) after capture in the Douro estuary, and after long-term depuration in an unpolluted laboratory environment. The results showed a significant decrease in EROD activity after 1 month and in bile metabolites after 4 months in captivity, with both maintaining reduced levels at 4 and 8 months depuration. Liver GST activity did not showed significant changes. This study provides evidence that Douro estuary waters contain bioavailable PAHs that can be associated with the induction of cytochrome P450, and that mullets have the ability to metabolise and eliminate PAHs. 相似文献
3.
The biotransformation of xenobiotics by microsomal cytochromes P450 is known to be pivotal in the effects of some compounds, and thought to be so for many. A knowledge of CYP gene diversity and CYP function and regulation in aquatic species is pursued, expecting that it will disclose mechanisms, allow predictions regarding species differences in susceptibility, and provide markers for exposure to xenobiotics. As well, it is hoped that such knowledge will provide clues to CYP endogenous functions, and to the origin and functional significance of CYP gene diversity. The knowledge of CYP in marine and other aquatic species is expanding rapidly. The diversity of CYP genes in non-mammalian vertebrates may approximate that in mammals. At present, cloning studies have identified members of gene families 1 to 4 have been cloned from one or more fish species. Where known, the gene structures of fish CYP genes are like those of mammalian homologues. Only one CYP1A gene has been identified in most fish species examined. Fish CYP1As, including multiple forms from recent divergence in some genera, have structural and catalytic properties more like CYP1A1, but also have properties that are 1A2-like, consistent with fish CYP1As representing the CYP ancestral to both CYP1A1 and CYP1A2. A number of genes cloned from several species have been classified in the 3A subfamily. Fish CYP3As catalyze steroid 6β-hydroxylase, and have other properties consistent with mammalian 3As. Recently identified CYP4 genes classify to novel subfamilies but apparently are homologues of mammalian CYP4 genes, and may act on similar substrates. The greatest diversity of fish CYP genes is in family 2; there are now six fish CYP2 subfamilies known. Four of these are novel subfamilies, although cladistic analysis suggests distinct relationships to mammalian CYP2 subfamilies. Heterologous expression and characterization of some of these CYP have identified similar functions among genes in different subfamilies. For example, fish CYP2Ns and CYP2Ps are related to mammalian CYP2Js, and CYP2P3 and CYP2J2 have strikingly similar functions as fatty acid epoxygenases and hydroxylases, with nearly identical regio- and enantioselectivity for metabolism of arachidonic acid. In addition to sequence and catalytic similarities, there also are indications that CYP regulation, tissue and cellular localization are similar between fish and mammals. Yet even in cases where orthology is strongly suggested, e.g. CYP1A, there appear to be taxonomic differences in active site structure suggesting potential differences in involvement of CYP1A in toxicity. In contrast to fish, CYP diversity and functions in aquatic invertebrates are poorly known. Investigators have identified novel gene families and subfamilies in crustaceans (CYP2L; CYP45), molluscs (CYP30, CYP10) and sponges (CYP38). CYP4C genes occur in crustaceans, molluscs and echinoderms, and a new subfamily (CYP4Y) in molluscs. The future? There is no doubt that new CYP will continue to be discovered in non-mammalian vertebrates; some (e.g. CYP51) can be predicted confidently. And, there is no doubt that the numbers known in invertebrates will expand greatly. In insects and C. elegans the numbers are very high, and even slime molds have 18 CYP genes. It is virtually certain that CYP genes with unique functions will be discovered. While the knowledge of CYP genes is increasing, knowledge of CYP function and regulation lag well behind. Technical approaches to speed the aquisition of such knowledge are available. The information will be essential to discern the role that CYP play in the disposition and toxicity of xenobiotics, during development as well as in adults. Yet, when such data are in hand, we may have to face the paucity of information on the diversity, function and regulation other enzymes, notably the glutathione S-transferases, glucuronyl transferases and sulfotransferases, in aquatic species. Discerning orthologous relationships among CYP genes, as well as those for phase II enzymes, could highlight gene lineages associated with conserved and endogenous functions. Understanding CYP endogenous functions, as well as their metabolism of xenobiotics, may reveal fully the ways that chemicals cause toxicity. [Support: Sea Grant NA46RG0470-R/P61, EPA R-829890, NIH ES07381]. 相似文献
4.
化学防御系统是生物抵御环境中化学胁迫和维持内环境稳态的重要机制,由长期进化出的一系列基因家族、蛋白和相关反应通路组成。该系统的核心组分包括:(1)受体和配体激活转录因子,其负责感知有毒化合物;(2)生物转化酶,其通过氧化、还原和结合反应使有毒物质的毒性降低,增加亲水性,使化合物更易排出;(3)外排转运蛋白,其负责将外源物质或生物转化产生的代谢物排出细胞;(4)抗氧化酶,其作用是保护细胞免受外部和内部产生的活性氧或自由基的伤害。本综述对动物化学防御系统的组成及作用机制进行概述,为研究海洋无脊椎动物的化学防御系统提供参考。 相似文献
5.
Relative potency of PAHs and heterocycles as aryl hydrocarbon receptor agonists in fish 总被引：6，自引：0，他引：6
The relative potency of polycyclic aromatic compounds as aryl hydrocarbon receptor (AhR) agonists in fish was determined using data on CYP1A induction or AhR binding for 74 polycyclic aromatic hydrocarbons (PAHs) and heterocycles in teleost, avian, or mammalian systems from 18 published papers. Each PAH was assigned a fish potency factor relative to the potency of 2,3,7,8-tetrachlorodibenzo-p-dioxin as an AhR agonist. Two and three ring unsubstituted PAHs were generally inactive in fish, avian, and mammalian systems. Benzo[k]fluoranthene and indeno[1,2,3-cd]pyrene were consistently the most potent PAHs, with fish potency factors of 0.001-0.002. Common structural features associated with higher potency PAHs included 4-6 rings containing fluoranthene or phenanthrene structures with an exposed bay region. These results show that PAHs can have similar potency as many dioxin-like PCBs, and AhR mediated toxicity should be considered in assessing the risks of PAHs in fish. 相似文献
6.
The aim of the present study was to investigate the responses of phase I and II biotransformation enzymes and levels of PAHs in the Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) collected from three sites at different distance from an oil refinery. Phase I enzyme activities as NAD(P)H-cyt c red, NADH ferry red, B(a)PMO and phase II as UDPGT, GST were measured in digestive gland while 16 PAHs (US-EPA) in whole soft tissue. An added value to the data obtained in the present study rely on the RDA analysis which showed close correlations between PAHs levels and phase I enzyme activities in mussels collected in front of the refinery. And again a significant spatial correlation between B(a)P levels and NADPH-cyt c red activities was observed using linear models. No differences among sites for B(a)PMO and phase II GST activities were observed, while the application of UDPGT as biomarkers requires further investigation. 相似文献
7.
Biological effects of environmental pollution, mainly related to presence of PAHs, were assessed in mussels Mytilus galloprovincialis caged in Priolo, an anthropogenically-impacted area, and Vendicari, a reference site, both located along the eastern coastline of Sicily (Italy). PAHs concentration and histopathological changes were measured in digestive gland tissues. Expression of cytochrome P4504Y1 (CYP4Y1) and glutathione S-transferase (GST), indicative of xenobiotic detoxification, and activity of catalase (CAT) as oxidative stress index, were evaluated.The results show a direct correlation between the high concentrations of PAHs in digestive glands of mussels from Priolo and the significantly altered activity of phase I (P < 0.001) and phase II (P < 0.0001) biotransformation enzymes, along with increased levels of CAT activity (P < 0.05). These findings show the enhancement of the detoxification and antioxidant defense systems. The mussel caging approach and selected biomarkers demonstrated to be reliable for the assessment of environmental pollution effects on aquatic organisms. 相似文献
8.
细胞系的构建在生物学领域具有重要意义,与其他动物(如昆虫)不同,经过数十年的发展,海洋无脊椎动物细胞研究仍停留在原代培养水平,没有建立起克隆化的永生细胞系。本文主要介绍了近年来海洋无脊椎动物细胞培养的研究进展,还介绍了一个基本原理,即海洋无脊椎动物永生细胞系的建立,可以通过捕获细胞的干性而实现。捕获细胞干性的第一条途径是利用转录因子等直接由体细胞制备海洋动物的诱导多能干细胞(iPS细胞)。第二条途径是获取海洋动物的成体干细胞(ASC)。这些细胞在维持机体活性、再生、无性克隆、体内增殖等方面起重要作用。当培养条件正确时,海洋无脊椎动物iPS细胞和ASC将会保持其原始状态,利用干细胞自我复制的固有能力、不分化地进行增殖而形成永生细胞系。 相似文献
9.
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
10.
Polycyclic aromatic hydrocarbons (PAHs) are contaminants increasing in the environment largely due to burning of fossil fuels. Our previous work identified a synergistic toxicity interaction in zebrafish embryos occurring when PAHs that are agonists for the aryl hydrocarbon receptor (AHR) co-occur with PAHs that are CYP1A inhibitors. This toxicity is mediated by the AHR2, and morpholino knockdown of CYP1A exacerbated toxicity. This study tested two hypotheses: (1) in the absence of functional CYP1A, metabolism of PAHs is shunted towards CYP1B1, which has been shown in mammals to produce more reactive metabolites of PAHs; alternatively, (2) CYP1B1 serves a protective role similar to CYP1A. We used a morpholino approach to knockdown CYP1B1 alone and in co-knockdown with CYP1A to determine whether we could alter deformities caused by synergistic toxicity of PAHs. CYP1B1 knockdown was not different from non-injected controls; nor were CYP1B1+CYP1A co-knockdown deformities different from CYP1A knockdown alone. These data suggest that CYP1B1 is not a significant factor in causing synergistic toxicity of PAHs, nor, in contrast to CYP1A, in providing protection. 相似文献
11.
Studies of xenobiotic metabolism and the regulation of enzyme systems for their metabolism (Phase I and II enzyme systems) require large numbers of animals, intensive use of experimental aquarium systems and in some instances can pose major problems when the compounds are scarce, expensive or too toxic to be disposed of easily. Previous studies have demonstrated the usefulness of isolated primary hepatocytes of fish for metabolic and enzyme induction studies. Functionally and structurally competent hepatocytes were isolated from juvenile plaice and after overnight acclimation to culture conditions, were exposed to a variety of PAHs for 24h. The levels of CYP1A1, phenol UDPGT and GST-A mRNAs were then estimated by slot blotting and hybridisation to their cDNA probes. The method enabled rapid and easy determination of the structure/activity relationships of these compounds as inducers of these key Phase I and II enzymes, and is potentially useful for screening large numbers of compounds both as an invitro toxicity test and for mechanistic studies. 相似文献
12.
Tommy L.F. Leung Kirsten M. Donald Devon B. Keeney Anson V. Koehler Robert C. Peoples Robert Poulin 《新西兰海洋与淡水研究杂志》2013,47(4):857-865
Parasites, in particular trematodes (Platyhelminthes: Digenea), play major roles in the population dynamics and community structure of invertebrates on soft‐sediment mudflats. Here, we provide a list of the 20 trematode species currently known to infect molluscs, crustaceans and polychaetes from Otago Harbour (New Zealand) soft‐sediment intertidal areas, as well as information on their transmission modes, life cycles, andknown ecological impacts. Several of the host‐parasite species combinations recorded here are reported for the first time. We also provide DNA barcodes, based on sequences of the cytochrome oxidase subunit l (CO1) gene, for 19 of the 20 trematode species, to facilitate future identification of these parasites in marine ecological studies. 相似文献
13.
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
14.
从基因水平探讨海洋鱼类对海洋藻毒素的去毒分子机理。采用RT-PCR法成功克隆了黄斑篮子鱼Siganus oramin肝脏I时相代谢酶细胞色素P450 1A(CYP1A)、II时相代谢酶alpha型谷胱甘肽S-转移酶(GSTA)和rho型谷胱甘肽S-转移酶(GSTR)、热休克蛋白70 (HSP70)、alpha 1型钠钾ATP酶(ATP1A1)及β-肌动蛋白(beta-actin, ACT)基因cDNA核心序列,序列分别长879 bp、582 bp、588 bp、660 bp、749 bp和554 bp。序列同源性分析发现,属鲈形目的黄斑篮子鱼CYP1A、GSTA和GSTR与同属鲈形目的牙鲆Paralichthys olivaceus、欧洲鲽Pleuronectes platessa、真鲷Pagrus major、鲤形目的斑马鱼Brachydanio rerio 相应氨基酸序列同源性较高,CYP1A和GSTA与非洲爪蟾(两栖类)、鸡(鸟类)、小鼠、大鼠和人(哺乳类)相应氨基酸序列同源性低,这可能与鱼类I、II时相去毒酶基因承担水环境毒素去毒代谢的特殊功能有关;而HSP70、ATP1A和β-肌动蛋白在鱼类、两栖类、鸟类、哺乳类中均有较高的同源性,这可能与这些基因在机体中承担的最基本的生命功能相关。应用半定量RT-PCR的方法,以β-肌动蛋白作为外参照,在指数期增长范围内分别得到了CYP1A、GSTA、GSTR、HSP70和ATP1A1 mRNA与β-肌动蛋白mRNA (%)的比值,确定黄斑篮子鱼肝脏去毒相关基因的组成型表达水平。其中,黄斑篮子鱼肝脏CYP1A、GSTA和GSTR基因组成型表达相对较高,HSP70和ATP1A1基因组成型表达相对较低,这可能与不同基因在黄斑篮子鱼海洋藻毒素去毒分子机理中承担的作用相关,为海洋藻毒素在海洋鱼类中的积聚及代谢去毒分子机制的研究提供了相关数据。 相似文献
15.
Francisco Barros Vanessa Hatje Maria Betnia Figueiredo Wagner Ferreira Magalhes Haroldo Silveira Drea Elissandro Soares Emídio 《Estuarine, Coastal and Shelf Science》2008,78(4):753-762
The structure of the benthic macrofaunal assemblages of the estuarine portion of Paraguaçu River, NE, Brazil, and its relationship with surface sediment characteristics (trace metals, PAHs, nutrients and grain size) and physical variables were investigated at ten stations on two contrasting occasions, summer (dry season) and winter (rainy season). A total of 1258 individuals (632 in winter and 626 in summer) and 62 taxa representing polychaetes, crustaceans, bivalves, echinoderms, bryozoans, sponges, cnidarians and cephalochordates were collected. Benthic assemblages in the upper estuary were unlike those in the lower estuary and a clear substitution of benthic taxa along the estuary was observed. Macrofaunal invertebrates in the low salinity region, composed of coarse sediments, were dominated by tellinids, venerids (bivalves), cirolanids (isopods), cyclopoids (copepods), and nereidids (polychaetes). While the high salinity region, composed of fine sediments, were dominated by nuculids (bivalves), cirratulids (polychaetes), and by amphiurids (ophiuroids). The Paraguaçu estuarine system is not severely affected by anthropogenic activities. In the great majority of the study sites, concentrations of trace metals and PAHs in the sediments were near background values. Nutrients values were also low. We formulated new models of taxon distribution and suggested detailed studies on the effects of salinity variation and studies using functional approaches to better understand the processes causing the spatial patterns in tropical estuarine benthic assemblages. 相似文献
16.
海洋无脊椎动物抗氧化酶研究进展 总被引：1，自引：0，他引：1
海洋无脊椎动物是重要的经济水产生物和海洋生态系统的主要组成者。抗氧化酶系统在其环境适应性以及非特异性免疫中发挥着重要作用。本文从海洋无脊椎动物主要抗氧化酶类的种类、结构、环境适应性、酶学、基因和蛋白研究等几个方面进行综述并对今后海洋无脊椎动物抗氧化酶的相关研究提出展望。 相似文献
17.
Despite the fact that BaP is a carcinogen, mammalian immunosuppressant, and ubiquitous aquatic pollutant, knowledge regarding the effects of BaP on the immune system of fish is still lacking. To begin to fill this gap, studies were conducted in medaka to examine the effects and mechanisms by which BaP exposure might alter host immunocompetence. Fish, exposed by IP injection of BaP (2-600 microg/g BW), were examined after 48 h for effects upon immune function and CYP1A expression/activity. Benzo[a]pyrene, at a concentration below that which increased levels of CYPIA expression/activity (2 microg BaP/g BW) suppressed lymphocyte proliferation. Concentrations of BaP at 20 and 200 microg/g BW. suppressed antibody-forming cell (AFC) numbers, superoxide production, and host resistance against bacteria. In contrast, exposure to the low affinity aryl hydrocarbon receptor (AhR) agonist, benzo[e]pyrene (BeP), neither induced CYP1A expression nor altered immune function. Given the lack of immunosuppressive effects produced by BeP, and the fact that exposure to the AhR antagonist (and CYP1A inhibitor) alpha-naphthoflavone (ANF) ameliorated the suppressive effects of BaP upon AFC numbers, the AhR pathway (including CYP1A-mediated production of reactive BaP metabolites) appears important in mediating BaP-induced immunotoxicity in fish, as in mammals. In the past, the medaka has proven a successful model for assessing carcinogenic agents. These studies have demonstrated its utility for also determining the immunosuppressive effects of an important aquatic contaminant. 相似文献
18.
Jee-Hyun Jung Young Sun Chae Ha Na Kim Moonkoo Kim Un Hyuk Yim Sung Yong Ha Gi Myung Han Joon Geon An Eunsic Kim Won Joon Shim 《Ocean Science Journal》2012,47(3):209-214
This study describes the spatial variation and the duration of the impacts from the Hebei Spirit oil spill using specific biochemical indices in resident benthic fish. Enzymatic activities and biliary PAHs metabolites were higher at the site closer to the spill area in four months after spill incident. Regarding our results of detoxification response, markers of Phase I followed a similar trend in accordance with levels of biliary metabolites, while markers of phase II and GST appeared relatively unchanged. 相似文献
19.
In the present study, long-term exposure to PCB resulted in marked induction of the xenobiotic biotransformation enzymes UDP glucuronosyl transferase, glutathione transferase and DT-diaphorase (measured as menadione reductase and DCPIP reductase) in rainbow trout 15 or 20 weeks post-injection during the experimental period. The increase of the phase II enzymes were paralleled by a marked increase of the glutathione reductase activity. These results indicate that prolonged exposure to PCB results in increased production of organo-radicals. 相似文献
20.
Pristane (2,6,10,14 -tetramethylpentadecane) occurs ubiquitously in the marine environment. This hydrocarbon may be of biogenic or petrogenic origin.1 Recently it has been shown that residual amounts of this branched alkane increased in marine organisms after an oil spill.2,3 The lack of data on the fate of pristane in fish, added with the fact that this compound was considered by some authors as a non-metabolisable substance in vertebrates, including man,4 led us to investigate the capability of fish to metabolise pristane. In this study, urinary and fecal excretion, tissue distribution and metabolism of 3H-pristane were analysed in Salmo gairdneri R. after a single intragastric dose. In addition to unchanged hydrocarbon, various labelled compounds have been isolated and identified in liver, bile, faeces, urine and surrounding water, demonstrating that pristane was first oxidised to alcohols (pristanol and pristane-diol) and to acid (pristanic acid). The elimination of these compounds occurred in the form of conjugated products (primarily glucuronides) as well as free metabolites. 相似文献