B UDC 厦门大学博硕士论文摘要库
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2 Using Biomarkers to Study PAHs Toxicological Effects on Marine Fish Lin Jianqing A Dissertation Submitted To Xiamen University In Partial Fulfillment of the Requirement For The Degree of PhD (Environmental Science) Advisors: Professor Hong Huasheng Completed in Nov Commencement in Nov II
3 PAH 1 PAH PAH PAH 1 PAH 16 PAH 4.55 g/l PAH 2.86 g/g CAT SODLPO PAH III
4 PAH CAT SOD MDA PAH PAH PAH PAH IV
5 Using Biomarkers to Study PAH Toxicological Effects on Marine Fish Lin Jianqing Abstract 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 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 V
6 levels 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 contamination and 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 VI
7 significantly 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 VII
8 VIII
9 PAH PAH 3.1 PAH PAH PAH PAH PAH PAH PAH PAH IX
10 PAH 6.1 CAT PAH SOD PAH LPO PAH EPA 16 PAH 157 PAH 158 GC/MS 159 X
11 Naphthalene Np Np Acenaphthylene Acy Acy Acenaphthene Ace Ace Fluorene F F Phenanthrene Ph Ph Anthracene An An Fluoranthene Fl Fl Pyrene Py Py Benzo(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 PAH EPA 16 PAH BCF Bioconcentration factor BSA Bovine Serum Albumin CAT Catalase, EC CBB Coomassie Brilliant Blue Kow n-octanol/water partition coefficient, / LPO lipid peroxidation, MDA malondialdehyde LPO MDA MFO mixed function oxidase, NOEC No-observed-effect-concentration, PAH, PAHs polycyclic aromatic hydrocarbons, XI
12 POPs persistent organic pollutants, QSPR quantitative structure-property relationships, SOD superoxidase dismutase, EC SPE Solid phase extraction TEP 1,1,3,3-tetraethoxy-propane TBA Thiobarbituric acid XII
13 PAH PAH Coastal Ocean estuarine ( 1993) trap 90 polycyclic aromatic hydrocarbons, PAH persistent organic pollutants, POPsUN-ECE, 2000; Vallack, et al, Clark, Pott 1920s-1930s (a) Phillips, s 1950s 1000 [legend1]: p1-2 [l2]:. POPs. 2000, 39(10):43 46 Vallack HW, et al. Controlling persistent organic pollutants-what next?. Environmental Toxicology and Pharmacology, 1998, 6: [legend3]: Clark, R.B., Marine Pollution, 3 rd ed., Clarendon Press, Oxford, [legend4]: Phillips DH. Fifty years of benzo(a)pyrene. Nature, 1983, 303(9):
14 1992 Bols NC, et al, 2001Vigan, Luigi, et al, 2002 Schoket Bernadette, et al, 2001; Autrup Herman, 2000; Pavanello Sofia, et al, 2000; Bir Anna, et al, (1) PAH PAH 170, PAH 50, PAH 4, PAH 2, PAH 2, PAH PAH Eisler, 1987 (2) PAH Kennish, 1997; 1992; 1998; Witt, 1995; Guzzelia, et al, 1996; Bernard, et al, 1996 PAH 1000 (Kennish, 2001) PAH -, 1992;, 1990 PAH Arkona PAH 11ng/g(dw) PAH 1900ng/g(dw) Warnow Oder Haff PAH 17,000 ng/g(dw) (Witt G, et al, 1999) Kim GB, et al 1999 Guinan, et al 2001 Readman, et al, 2002 PAH PAH - [legend5]: P62 [legend6]: Bols NC, Brubacher JL, Ganassin RC, Lee LEJ. Ecotoxicology and innate immunity in fish. Developmental and Comparative Immunology,2001,25(8-9): [legend7]: Vigan, Luigi; Camoirano, Anna; Izzotti, Alberto; D Agostini, Francesco; Polesello, Stefano; et al. Mutagenicity of sediments [legend8]: Schoket, Bernadette; Papp, Gizella; L vay, 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 2
15 PAH PAH Pereira 1999 PAH Richardson BaySan Pablo Bay PAH PAH PAH ng/g.dw PAH ng/g.dw(Tang, et al, 1989; Lu, et al,1990) PAH PAH 877.2ng/g.dw (Ma, et al, 2001) PAH, 2000, 2001; Liu M, 2000, 2001, 2000; Bixian M, et al, 2001 PAH PAH (Kennish, 2001) (3) PAH PAH PAH PAH 120 g/g 60 PAH PAH 160 ng/g McLeese, et al, 1985 PAH ng/g PAH ng/g(windor, et al, 1979) PAH PAH PAH 1.1a 1.1b PAH PAH PAH PAH [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): 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 3
16 1987 biomarker indicators signaling events in biological system or samples, Fossi 1994 Depledge 1993 [legend25]: [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, pp Berlin: Springer-Verlag. 4
17 Degree papers are in the Xiamen University Electronic Theses and Dissertations Database. Full texts are available in the following ways: 1. If your library is a CALIS member libraries, please log on and submit requests 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.
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