水体Cu～（2+）对中华螯蟹（Eriocheir sinensis）毒性作用机制的研究
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摘要
近年来,我国的虾蟹养殖业发展迅速,形成了规模化生产,虾蟹养殖业已成为我国农民脱贫致富的重要产业。随着生产的发展,养殖用水的污染也在不断的加剧,其中除了受工业、农业和生活污水的影响之外,生产中大量使用硫酸铜等铜类化合物进行病害防治和清塘灭藻也已成为重要的水污染原因之一。2003年度的《中国渔业生态环境状况公报》强调指出我国近岸海域、江河、湖泊的渔业水域中,Cu~(2+)的污染超标程度仍在增加(以GB11607-89国家渔业水质标准中Cu~(2+)浓度不超过0.01mg/l为标准),特别是杭州湾、长江口和舟山近海平均值竟达0.07mg/l。Cu~(2+)是我国渔业水域环境中超标最严重的重金属。因此,自然水域中的经济甲壳动物的生长和发育受到Cu~(2+)胁迫的可能性充分存在。水污染的结果导致虾蟹成活率降低和养殖成本提高。同时,受Cu~(2+)等重金属污染后的产品出口创汇深受影响。因此,本项目选择水环境中重要的污染元素Cu~(2+)并深入开展Cu~(2+)对经济虾蟹动物生长的营养和毒性作用的研究,具有重要的现实意义和理论价值。
微量元素Cu~(2+)是甲壳动物重要的营养素,是合成血蓝蛋白的必需成分之一,并对多种生长发育相关的生物酶的组成和功能有着重要意义。然而,过量的Cu~(2+)又会对上述蛋白和生物酶产生毒害,且这些危害可以反映在生物体、细胞或分子水平上。可见,Cu~(2+)与甲壳动物的生长发育关系密切。虽然有报道水体Cu~(2+)对中华绒螯蟹的毒性作用,但大都集中在估算半致死浓度,缺乏Cu~(2+)对中华绒螯蟹毒性作用机制的研究,特别是针对X-器官窦腺复合体(XO-SG)和Y-器官(YO)的结构、功能影响的直接研究极少,至今无法从机理上解释Cu~(2+)影响甲壳动物生长的现象。
本研究以我国最重要的经济虾蟹类养殖品种之一中华绒螯蟹作为研究对象,综合形态学、生理学、生物化学、内分泌学、毒理生态学的方法和技术,辅以电镜、生化酶学、免疫检测、微量元素分析等手段,以期阐明水体Cu~(2+)对中华绒螯蟹的毒性作用机制。这不仅对丰富重金属对甲壳动物毒性作用的机制具有重要的理论价值,而且可以为健康养殖和水质监测提供参考数据,并对其它重金属类相应的研究建立成熟的方法。研究结果与结论如下:
In recent years, aquaculture industry of crustacean, especially shrimps and crabs, has shown rapid expansion in China, which has been developed into a major industry. However, effluent from human activities and application of Copper sulfate to control algal in crustacean ponds resulted in Copper (Cu~(2+)) pollution that stunts the further growth of this aquaculture industry. The 2003 Report on State of the Fishery Eco-environment in China emphasized that Cu~(2+) is the heavy metal that exceeds most evidently the Standards of the Water Quality in the Fishery Industry (GB11607-89). In certain regions severity of Cu~(2+) contamination is still increasing, e.g. in the Hangzhou bay, piscatorial areas of Yangtse Rivers and the coast of Zhoushan. The average Cu~(2+) level of these waters is 0.07 mg/1, being 7 folds higher than that stipulated of GB 11607-89. Therefore, the Cu~(2+) toxicity on crustacean of fisheries is of primary concern. As Cu~(2+) pollution can be severely limiting factors to future crustacean culture potentially, studies on its influence on E. sinensis are of great significance in elucidating the physiological alterations of animals and establishing methods for safe aquaculture. In the present study the toxic mechanism of water-borne Cu~(2+) on E. sinensis is investigated in detail. This work is believed to have both important realistic and theoretic significances.Cu~(2+) is one of the important nutrient elements, being essential to synthesize hemocyanin in crustaceans. Cu~(2+) is required for normal function of many proteins, including most biological enzymes that are necessary for normal growth and development. However, excess Cu~(2+) is toxic and can causes a variety of adversely affects to crustaceans on biological functions from molecule level to cell level. Although many works on the toxic effects of Cu~(2+) on E. sinensis have already been reported, they almost focus on the evaluation of LC_(50) of pollutants rather than on the mechanism of toxic effects of water-borne Cu~(2+), esp. the toxic impacts on the structure and function of X organ-sinus gland complex and the Y-organ, two most important organs for ecdysis and growth of crustaceans. So far the mechanism of Cu~(2+) on the growth of crustaceans has still not been well understood.
In the present study, the effects of water-borne Cu~(2+) the Chinese mitten crab E. sinensis have been conducted by integrated morphologic, physiologic, biochemical, endocrinologic, ecotoxicologic approaches and equipments of electric microscopic, biochemical enzymes and trace element analysis. The aim of this work is to clarify the mechanism of toxic effects of Cu~(2+) on E. sinensis and it must not only be of important theoretic value on the toxic effects of heavy metal on crustaceans, but also provide useful references on the healthy aquaculture and the effective evaluation of water quality. The main results in this thesis can be summarized as follows:1 Effects of water-borne Cu~(2+) on molting, growth and survival in E. sinensisThe acute toxic effects of water borne Cu~(2+) on the molting rate, the weight gain and the survival rate in E. sinensis was conducted between I stage and 12-month-old juveniles. The 24, 48, 72 and 96 h LC_(50)s of Cu~(2+) on I stage juveniles (0.020 ± 0.01g) were 0.70, 0.43, 0.33, and 0.22 mg/l, and those on the 12-month-old animals (3.34 ± 0.26 g) were 18.20, 10.23, 9.12 and 8.51 mg/1, respectively. Results suggested that the survival rate of all I stage juvenile groups were higher than 50%, and furthermore it decreased in accordance with the increase of water borne Cu~(2+) level (from 0.00 to 0.01, 0.02, 0.03, 0.05 and 0.08 mg/l). The similar decrease tendency was also observed in the 12-month-old E. sinensis. In addition, the molting rate, weight gain and the survival rate of the 12-month-old animals decreased stably in accordance with the increase of Cu~(2+) level in the Cu~(2+) treated groups (0.01, 0.05, 0.10, 0.50, 1.00, 2.50 mg/l) groups. The survival rates in all groups studied were higher than 50%. Although the survival rate in
微量元素Cu~(2+)是甲壳动物重要的营养素,是合成血蓝蛋白的必需成分之一,并对多种生长发育相关的生物酶的组成和功能有着重要意义。然而,过量的Cu~(2+)又会对上述蛋白和生物酶产生毒害,且这些危害可以反映在生物体、细胞或分子水平上。可见,Cu~(2+)与甲壳动物的生长发育关系密切。虽然有报道水体Cu~(2+)对中华绒螯蟹的毒性作用,但大都集中在估算半致死浓度,缺乏Cu~(2+)对中华绒螯蟹毒性作用机制的研究,特别是针对X-器官窦腺复合体(XO-SG)和Y-器官(YO)的结构、功能影响的直接研究极少,至今无法从机理上解释Cu~(2+)影响甲壳动物生长的现象。
本研究以我国最重要的经济虾蟹类养殖品种之一中华绒螯蟹作为研究对象,综合形态学、生理学、生物化学、内分泌学、毒理生态学的方法和技术,辅以电镜、生化酶学、免疫检测、微量元素分析等手段,以期阐明水体Cu~(2+)对中华绒螯蟹的毒性作用机制。这不仅对丰富重金属对甲壳动物毒性作用的机制具有重要的理论价值,而且可以为健康养殖和水质监测提供参考数据,并对其它重金属类相应的研究建立成熟的方法。研究结果与结论如下:
In recent years, aquaculture industry of crustacean, especially shrimps and crabs, has shown rapid expansion in China, which has been developed into a major industry. However, effluent from human activities and application of Copper sulfate to control algal in crustacean ponds resulted in Copper (Cu~(2+)) pollution that stunts the further growth of this aquaculture industry. The 2003 Report on State of the Fishery Eco-environment in China emphasized that Cu~(2+) is the heavy metal that exceeds most evidently the Standards of the Water Quality in the Fishery Industry (GB11607-89). In certain regions severity of Cu~(2+) contamination is still increasing, e.g. in the Hangzhou bay, piscatorial areas of Yangtse Rivers and the coast of Zhoushan. The average Cu~(2+) level of these waters is 0.07 mg/1, being 7 folds higher than that stipulated of GB 11607-89. Therefore, the Cu~(2+) toxicity on crustacean of fisheries is of primary concern. As Cu~(2+) pollution can be severely limiting factors to future crustacean culture potentially, studies on its influence on E. sinensis are of great significance in elucidating the physiological alterations of animals and establishing methods for safe aquaculture. In the present study the toxic mechanism of water-borne Cu~(2+) on E. sinensis is investigated in detail. This work is believed to have both important realistic and theoretic significances.Cu~(2+) is one of the important nutrient elements, being essential to synthesize hemocyanin in crustaceans. Cu~(2+) is required for normal function of many proteins, including most biological enzymes that are necessary for normal growth and development. However, excess Cu~(2+) is toxic and can causes a variety of adversely affects to crustaceans on biological functions from molecule level to cell level. Although many works on the toxic effects of Cu~(2+) on E. sinensis have already been reported, they almost focus on the evaluation of LC_(50) of pollutants rather than on the mechanism of toxic effects of water-borne Cu~(2+), esp. the toxic impacts on the structure and function of X organ-sinus gland complex and the Y-organ, two most important organs for ecdysis and growth of crustaceans. So far the mechanism of Cu~(2+) on the growth of crustaceans has still not been well understood.
In the present study, the effects of water-borne Cu~(2+) the Chinese mitten crab E. sinensis have been conducted by integrated morphologic, physiologic, biochemical, endocrinologic, ecotoxicologic approaches and equipments of electric microscopic, biochemical enzymes and trace element analysis. The aim of this work is to clarify the mechanism of toxic effects of Cu~(2+) on E. sinensis and it must not only be of important theoretic value on the toxic effects of heavy metal on crustaceans, but also provide useful references on the healthy aquaculture and the effective evaluation of water quality. The main results in this thesis can be summarized as follows:1 Effects of water-borne Cu~(2+) on molting, growth and survival in E. sinensisThe acute toxic effects of water borne Cu~(2+) on the molting rate, the weight gain and the survival rate in E. sinensis was conducted between I stage and 12-month-old juveniles. The 24, 48, 72 and 96 h LC_(50)s of Cu~(2+) on I stage juveniles (0.020 ± 0.01g) were 0.70, 0.43, 0.33, and 0.22 mg/l, and those on the 12-month-old animals (3.34 ± 0.26 g) were 18.20, 10.23, 9.12 and 8.51 mg/1, respectively. Results suggested that the survival rate of all I stage juvenile groups were higher than 50%, and furthermore it decreased in accordance with the increase of water borne Cu~(2+) level (from 0.00 to 0.01, 0.02, 0.03, 0.05 and 0.08 mg/l). The similar decrease tendency was also observed in the 12-month-old E. sinensis. In addition, the molting rate, weight gain and the survival rate of the 12-month-old animals decreased stably in accordance with the increase of Cu~(2+) level in the Cu~(2+) treated groups (0.01, 0.05, 0.10, 0.50, 1.00, 2.50 mg/l) groups. The survival rates in all groups studied were higher than 50%. Although the survival rate in
引文
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