污染物对海洋生物毒性效应的早期预警和生物标志物指示作用研究是当今海洋生态毒理学研究的热点和前沿。本论文以湄洲湾海域和厦门西海域养殖区为研究区域，采用现代分析技术，从分子水平上系统研究了多环芳烃（PAH）在海水、沉积物、养殖鱼类中的含量分布；通过现场实验，实验室暂养、投毒和恢复实验系统测定了养殖鱼类的肝脏、胆汁、鳃、全血等不同器官、组织的生化指标及养殖鱼类肝脏、胆汁的化学指标，探讨了这些指标对水环境多环芳烃污染的生物指示作用。取得如下新的成果：1首次系统研究了多环芳烃在湄洲湾海水、沉积物的含量分布，采用PAH型式的分子指纹方法分析了水体、沉积物中PAH的来源特征；指出：湄洲湾海水的多环芳烃主要来...Polycyclic aromatic hydrocarbons (PAH) are priority trace organic pollutants in the CLRTAP (Convention on Long-Range Transboundary Air Pollution) POPs (persistent organic pollutants) protocol. Both laboratory experiments and field surveys unequivocally demonstrated that PAH adversely affect estuarine and marine organisms. PAH are implicated in the development of lesions and tumors in fish，they pro...学位：工学博士院系专业：海洋与环境学院环境科学研究中心_环境科学学号：B19993400

林建清

Xiamen University Institutional Repository

10384     B9934003  UDC                :  2002.11.3    2002.12.4   2002 12 3厦门大学博硕士论文摘要库IIUsing Biomarkers to Study PAHs’ Toxicological Effects on Marine Fish Lin Jianqing A Dissertation Submitted To Xiamen University In Partial Fulfillment of the Requirement ForThe Degree of PhD (Environmental Science) Advisors: Professor Hong Huasheng Completed in Nov. 2002 Commencement in Nov. 2002 厦门大学博硕士论文摘要库IIIPAH1 PAHPAHPAH1PAH 16 PAH 4.55 g/L PAH 2.86 g/g234 1 1 1 11 23 15 CATSOD LPO PAH厦门大学博硕士论文摘要库IVPAH CAT SOD MDAPAH PAHPAHPAH厦门大学博硕士论文摘要库VUsing Biomarkers to Study PAH Toxicological Effects on Marine Fish Lin Jianqing AbstractPolycyclic aromatic hydrocarbons (PAH) are priority trace organic pollutants in the CLRTAP (Convention on Long-Range Transboundary Air Pollution) POPs (persistent organic pollutants) protocol. Both laboratory experiments and field surveys unequivocally demonstrated that PAH adversely affect estuarine and marine organisms. PAH are implicated in the development of lesions and tumors in fish they produce biochemical disruptions and cell damage that lead to mutations, developmental malformations and cancer. Although many marine organisms do not exhibit acute responses to PAH’ exposure, but suffer sublethal effects. Impacts on population and community may not be detected until years after initial PAH’ exposure. Therefore, it is important to develop criteria to indicate PAH’ pollution status in the marine environments. Biomarkers are promising tools for assessing the exposure to toxicants, and predicting the likely response. Since the complexity of the marine environments, the toxic effects to biota are very complicated; therefore, the use of biomarkers for assessing marine environments is still rather ambitious. The aim of this study is to examine PAH’ distribution in water and sediment, analyze their sources, and try to develop a set of biomarkers to indicate PAH’ pollution level in the following study areas.  The Meizhou Bay, located in the mid-east coast of Fujian, China, is a semi-enclosed narrow bay stretching deep into the inland with favorable hydrological circulation conditions and designed as one of the four key deepwater transshipment ports in China. With the construction of ports, oil-refinery works and power industry in the coastal area; PAH’ pollution is an potential environmental risk factor in this marine environment, but no data of PAH level in this marine environment were examined before. Western Xiamen Harbor , located in Xiamen, China with harbor rapidly development; PAH’ 厦门大学博硕士论文摘要库VIlevels in water, suspended particulate matter, pore water and sediment samples were studied before, but no data on the maricultural sites in this area were examined yet. The field surveys and lab experiments were designed to study the distribution and source of PAH’ pollution in water, sediment; the PAH’ concentration and distribution in marine fish organ (of the fish Lateolabrax japonicus and Pagrosomus major); the PAH’ BCF factors of liver and bile in marine fish (Lateolabrax japonicus and Pagrosomus major); the relationship between PAH’ exposure and biochemical response (CAT, SOD, LPO) and the metabolic rate of naphthalene and pyrene in fish bile. The following results were obtained:    1 In Meizhou Bay:    (1) The source of PAH in surface seawater was mainly from crude oil petrogenic contaminationand the source of PAH in sediments was mainly from fuel-combustion (pyrogentic).    (2) Refer to the extrapolated safe levels developed by various countries and international organizations for aquatic organisms exposed to PAH in water to assess aquatic quality for maricultural purpose, and the results showed that PAH’ pollution in seawater was already a threat for its maricultural use.   (3) PAH’ levels in maricultural site were much higher than that in the adjacent non-maricultural site, and it showed that mariculture was one of the contributors of marine PAH’ pollution in seawater.     2  Western Xiamen Harbor:     (1) PAH’ levels in surface seawater and sediment of the survey site were quite severe. PAH (for 16 EPA priority PAH) concentration in surface seawater was 4.55 g/L, and 2.86 g/g in surface sediment.    (2) The survey maricultural site was quite polluted, and it was not suitable for maricultural purpose.   3 The results showed that fish bile could concentrate parent PAH, and the PAH’ concentration correlated to the PAH’ pollution levels in seawater, thus, it could be used as a biomarker to indicate PAH’ pollution levels in seawater. The metabolism of naphthalene and pyrene was also studied, and the results showed that pyrene is more easily metabolized than naphthalene.    4 The bioconcentration and clearance of PAH in fish liver were systematically studied. The results showed that: fish liver could strongly concentrate PAH (BCF>500); and PAH’ level in fish liver was 厦门大学博硕士论文摘要库VIIsignificantly related to PAH pollution level in seawater, thus, PAH’ level in fish liver could be used to indicate PAH’ pollution levels in seawater.   5 The distribution of CAT activities in fish organs was studied, and the activities in fish organs were shown as the following sequence: liver, whole blood >gill>>mussel. CAT activities in liver and gill were more sensitive than that in whole blood exposed to PAH. In the experiment conditions, CAT activities in fish liver were reversible to PAH exposure, CAT activities in fish liver may be used as an early warning indicator of PAH exposure.   6 SOD activities in fish liver were positive relevant to PAH levels in fish bile and seawater and SOD activities in fish liver were reversible, therefore, it could also be used as a biomarker and an early warning indicator of PAH pollution.   7 MDA (malondialdehyde) concentrations were used to indicate lipid peroxidation level. The higher MDA concentrations, the stronger lipid peroxidation. The results showed that:   (1) MDA concentrations in gill and whole blood were higher than that in liver, and were negative relevant to SOD and CAT activities.   (2) MDA concentration in fish liver was reversible to the exposure of PAH pollution level and positive relevant to PAH concentration, thus, and it also could be used as biomarker and early warning indicator to indicate PAH pollution level.    In summary, PAH’ concentration in fish liver and bile, 1-pyrenol concentration in fish bile could be used as biomarkers to indicate PAH’ pollution level in seawater. CAT, SOD and LPO levels in fish liver could also be used as biomarkers and early warning indicators to indicate PAH’ pollution level in seawater. But biochemical factors (such as CAT, SOD, LPO ) were also affected by other chemical pollutants, therefore, it is better to combine chemical factors (concentration level) and biochemical factors to indicate the PAH’ pollution in marine environment.   Keywords: biomarker, PAH, ecotoxicological effects to marine fish, antioxidant enzymes厦门大学博硕士论文摘要库VIII2002 11厦门大学博硕士论文摘要库IX1.1   1       1.2   3 1.3  PAH  7 1.4  9 2.1  16 2.2  17 2.3  19 2.4 212.5  22 2.6  23  PAH3.1 PAH   33 3.2 PAH   48 3.3  70 PAH PAH4.1  73 4.2 PAH PAH  75 4.3 PAH PAH  79 15.1  90 5.2 1 1  96 厦门大学博硕士论文摘要库X5.3 1 97PAH6.1 CAT PAH 1006.2 SOD PAH  114 6.3 LPO PAH 121136138EPA 16 PAH 157PAH 158 GC/MS  159 厦门大学博硕士论文摘要库XI161 2Naphthalene Np NpAcenaphthylene Acy Acy Acenaphthene Ace AceFluorene F FPhenanthrene Ph PhAnthracene An AnFluoranthene Fl FlPyrene Py PyBenzo(a)anthracene  B( )An BaA (a)Chrysene  Chry Chry Benzo(b)fluoranthene B(b)Fl BbF (b)Benzo(k)fluoranthene B(k)Fl BkF (k)Benzo(a)pyrene B( )Py BaP (a)Indeno(1,2,3-cd)pyrene I(1,2,3-cd)Py IPy (1,2,3-cd)Dibenzo(a,h)anthracene dB(a,h)An dBA (a,h)Benzo(ghi)perylene  B(ghi)Pe BPe (ghi)PAH PAHs 16 EPA PAH 16 PAHEPA 16 PAHBCF Bioconcentration factorBSA Bovine Serum AlbuminCAT  Catalase, EC1.11.1.6 CBB Coomassie Brilliant BlueKow n-octanol/water partition coefficient, /LPO  lipid peroxidation, MDA malondialdehyde LPO MDAMFO mixed function oxidase, NOEC No-observed-effect-concentration, PAH, PAHs polycyclic aromatic hydrocarbons, 厦门大学博硕士论文摘要库XIIPOPs persistent organic pollutants, QSPR quantitative structure-property relationships, SOD superoxidase dismutase, EC. 1.15.1.1 SPE  Solid phase extractionTEP 1,1,3,3-tetraethoxy-propaneTBA Thiobarbituric acid厦门大学博硕士论文摘要库PAHPAH1.11.1.1 Coastal Ocean estuarine 80.5 30( 1993)trap 90polycyclic aromatic hydrocarbons, PAH persistent organic pollutants, POPs UN-ECE, 2000; Vallack, et al, 19981.1.2 Clark , 19921775 Pott1920s-1930s(a)Phillips, 1983 1950s1950s1000 [legend1]: ..1993p1-2  [l2]: . POPs. 2000, 39(10):43 46Vallack HW, et al. Controlling persistent organic pollutants-what next?. Environmental Toxicology and Pharmacology, 1998, 6:143175  [legend3]: Clark, R.B., Marine Pollution, 3rd ed., Clarendon Press, Oxford, 1992.  [legend4]: Phillips DH. Fifty years of benzo(a)pyrene. Nature, 1983, 303(9): 468-472 厦门大学博硕士论文摘要库 21992Bols NC, et al, 2001 Vigan , Luigi, et al, 2002Schoket Bernadette, et al, 2001; Autrup Herman, 2000; Pavanello Sofia, et al, 2000; BirAnna, et al, 2002     1.1.3(1) PAH 2.3 105PAH 170,000 73.90 PAH 50,000 21.74 PAH 4,400 1.91 PAH 2,940 1.28 PAH 2,7001.17 PAHPAH Eisler, 1987(2) PAH Kennish, 1997; 1992; 1998; Witt, 1995; Guzzelia, et al, 1996; Bernard, et al, 1996PAH 1000 (Kennish, 2001) PAH- , 1992; , 1990PAHArkona PAH 11ng/g(dw) PAH1900ng/g(dw) Warnow Oder Haff PAH17,000 ng/g(dw) (Witt G, et al, 1999)  Kim GB, et al 1999 Guinan, et al 2001 Readman , et al, 2002 PAHPAH - [legend5]: ..1992P62  [legend6]: Bols NC, Brubacher JL, Ganassin RC, Lee LEJ. Ecotoxicology and innate immunity in fish.  Developmental and Comparative Immunology,2001,25(8-9): 853-873    [legend7]: Vigan , Luigi; Camoirano, Anna; Izzotti, Alberto; D Agostini, Francesco; Polesello, Stefano; et al. Mutagenicity of sediments  [legend8]: Schoket, Bernadette; Papp, Gizella; Lvay, Katalin; Mrackov , [legend9]: Eisler R. Polycyclic Aromatic Hazards to Fish, Wildlife, and Invertebrates:  [legend10]: Kennish, M.J., Ecology of Estuaries: Anthropogenic Effects, CRC  [legend11]: Kennish, MJ, Ed., Practical Handbook of Estuarine and Marine Pollution,  [legend12]: .. [legend13]: Witt G, Trost E. Polycyclic aromatic hydrocarbons (PAH) in  [legend14]: Kim GB, Maruya KA, Lee RF, et al. Distribution and sources of  [legend15]: Guinan J, Charlesworth M, Service M, et al. Sources and geochemical 厦门大学博硕士论文摘要库 3PAHPAH Pereira 1999PAHRichardson Bay San Pablo BayPAH PAHPAH 150 330ng/g.dwPAH 50 150ng/g.dw(Tang, et al, 1989; Lu, et al,1990) PAHPAH877.2ng/g.dw (Ma, et al, 2001)PAH , 2000, 2001; Liu M, 2000, 2001 ,2000;  Bixian M, et al, 2001PAH PAH(Kennish, 2001)(3) PAHPAHPAH PAH 120 g/g 60 PAHPAH 160 ng/g McLeese, et al, 1985PAH 200 870ng/g PAH 18 160 ng/g(Windor, et al, 1979) PAHPAHPAH 1.1a 1.1bPAH PAHPAH PAH1.21.2.1  [legend16]: Pereira WE, Hostettler FD, Luoma SN, et al. Sedimentary record of anthropogenic and biogenic polycyclic aromatic hydrocarbons in San Francisco Bay, California. MAR CHEM 64 (1-2): 99-113 FEB 1999  [legend17]: Tang Yunqian, Alkane and aromatic hydrocarbons in sediments in the sea area near the Antarctic Peninsula, In: symposium on cooperation programs between  [legend18]: Lu Bin, Tang Yunqian, et al. Distribution features of dissolved hydrocarbons in water mass of  [legend19]: Ma M, Feng Z, Guan C, et al. DDT, PAH and PCB in sediments from the intertidal zone of the Bohai Sea  [legend20]:  .  .  [legend21]:  .  [legend22]: Kennish, MJ, Ed., Practical Handbook of Estuarine and Marine Pollution, CRC Press, Boca Raton,FL,  [legend23]: McLeese, D.W., Ray, S., and Burridge, L.E., Accumulation of polynuclear aromatic  [legend24]: Windor JG, and Hites RA. Polycyclic aromatic hydrocarbons in Gulf of Maine sediments and Nova 厦门大学博硕士论文摘要库 41987biomarkerindicators signaling events in biological system or samples , 199790 Fossi 1994Depledge 1993 [legend25]: .. 1997 9 122-35  [legend26]: Fossi MC, and Leonzio C(ed) (1994) Nondestructive Biomarker in Vertebrate,Boca Raton, FL: Lewis [legend27]: Depledge MH, Amaral-Mendel JJ, Daniel B, et al. The conceptual basis of the biomarker approach. In Peakall DB and Shugart RL(eds.) Biomarkers. Research and application in the assessment of Environmental Health, pp15-29. Berlin: Springer-Verlag.  厦门大学博硕士论文摘要库Degree papers are in the “Xiamen University Electronic Theses and Dissertations Database”. Fulltexts are available in the following ways: 1. If your library is a CALIS member libraries, please log on http://etd.calis.edu.cn/ and submitrequests online, or consult the interlibrary loan department in your library. 2. For users of non-CALIS member libraries, please mail to etd@xmu.edu.cn for delivery details.厦门大学博硕士论文摘要库

Using Biomarkers to Study PAHs’ Toxicological Effects on Marine Fish

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Using Biomarkers to Study PAHs’ Toxicological Effects on Marine Fish

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