氯氰菊酯对鲫鱼的毒理效应研究
摘要
氯氰菊酯是一种拟除虫菊酯杀虫剂，随着近年来在农业生产上广泛应用，在环境中的污染日趋严重。本论文以鲫鱼为试验动物，应用生物化学、细胞生物学等技术手段，研究了氯氰菊酯对鲫鱼的急性毒性、在鲫鱼体内的蓄积释放规律、对鲫鱼的免疫系统、抗氧化防御体系以及遗传效应的影响，从整体生物、组织细胞水平和分子水平多层次地评价了氯氰菊酯对鲫鱼的毒理效应。
研究结果表明，氯氰菊酯对平均体长为14．0±1．0cm，平均体重为65．0±5．0g的鲫鱼96hLC_(50)值为18．0μg·L~(-1)，根据农药毒性分级标准，氯氰菊酯对鲫鱼属高毒类农药。
根据急性毒性试验结果，选择1／2 96hLC_(50)以下浓度为试验浓度系列，即1μg·L~(-1)，2μg·L~(-1)，3μg·L~(-1)，5μg·L~(-1)和8μg·L~(-1)对鲫鱼进行连续暴露染毒，分析氯氰菊酯的低剂量连续暴露对鲫鱼的生理生化和遗传指标的影响，结果如下：
1)氯氰菊酯经20-25d可在鲫鱼体内达到蓄积平衡，肌肉和肝脏组织中的最大富集系数分别为3．950和2．155，肌肉组织的蓄积能力小于肝脏组织。在氯氰菊酯的释放过程中，肌肉组织的释放快于肝脏组织。
2)经氯氰菊酯30d连续暴露后鲫鱼肝脏发生了明显的改变，但脏器系数未有显著的变化；而鲫鱼的脾脏无论是解剖形态，还是脏器系数与对照组相比均无明显差异。氯氰菊酯对鲫鱼溶菌酶的影响表现为低浓度组(2μg·L~(-1)-5μg·L~(-1))溶菌酶呈现先升高后降低的趋势，而高浓度组(8μg·L~(-1))则相反，为先降低后升高。但随着暴露时间的延长，各试验组在20-25d时恢复至正常水平，血清中溶菌酶的含量与对照组相比无明显差异，至试验结束时(第30d)各浓度组溶菌酶的含量稳定，变化不大。
3)鲫鱼在1．0μg·L~(-1)氯氰菊酯连续暴露30d后，其肌肉和肝脏中的GSH含量和GST、CAT、MAO的酶活性与对照组相比均未发生显著的差异(P>0．05)，而2．0μg·L~(-1)以上浓度的氯氰菊酯连续暴露对鲫鱼肌肉和肝脏中的GSH含量和GST、CAT、MAO活性均产生了一定程度的影响：(a)氯氰菊酯连续暴露下鲫鱼肌肉和肝脏中的GSH含量和GST活性的变化规律为先升高后降低，且肝脏中的变化要大于肌肉中变化。在试验结束时，除1．0μg·L~(-1)试验组外，其余试验组GSH含量和对照组相比均存在一定的差异；而2．0μg·L~(-1)及以上试验组GST活性在试验结束时与对照组相比差异显著(P<0．01)，低至对照组的15．71％--67．31％。(b)CAT活性的变化规律为先降低后升高，肝脏中的变化幅度要大于肌肉，在试验结束时除1．0μg·L~(-1)试验组外，其余试验组CAT活性与对照组相比均有一定的变化。肌肉中的CAT活性与对照组相比最大降幅可达17．8％，肝脏组织CAT最大降幅可达51．86％-69．64％(P<0．01)。(c) MAO的变化规律为先升高后降低再恢复，在试验结束时除1．0μg·L~(-1)试验组外，其余试验组MAO活性与对照组相比均有一定的变化。
4)3．0μg·L~(-1)以上浓度的氯氰菊酯连续暴露可诱导鲫鱼外周血红细胞产生微核，微核的发生率随暴露浓度的升高和暴露时间的延长具有明显的浓度-效应和时间-效应关系。
研究结果显示，2．0μg·L~(-1)以上浓度的氯氰菊酯可对鲫鱼的生理机能产生一定的影响，这种影响在一定程度上关系到鲫鱼的养殖品质。因此为了鲫鱼的养殖和食用安全，在养殖水环境中氯氰菊酯的浓度应严格控制于1．0μg·L~(-1)以下。
Cypermethrin which was a kind of pyrethroids caused severe contamination to the environment with its wide use in agriculture in recent years . This master paper used Carassius auratus Linn. as experiment animal, applied biochemiscalogy, cell biology etc. technology , studied the influences produced by cypermethrin on Carassius auratus Linn, from acute toxicity trials , accumulation and release rules , immune system , antioxidize system , and heredity effect, from the whole living, organ and cell level and molecule level evaluated the satey of cypermethrin .
The results of acute toxicity revealed that 96hLC_(50) of cypermethrin was 18.0μg·L~(-1) for Carassius auratus Linn. with mean body weight 65.0g±5.0g and mean body length 14.0±1.0cm. According to the methods of biological detect, cypermethrin was a high toxic pesticide for Carassius auratus Linn..
Based on the acute toxicity trial, the following concentration series below 1/2 96hLC_(50) that were 1μg·L~(1),2μg·L~(1),3μg·L~(1) ,5μg/L and 8μg·L~(1) were established to infect Carassius auratus Linn.. Analyzing continual exposure to low dosage cypermethrin for the impact of physiochemical and hereditary of Carassius auratus Linn.. We get a conclusion that:
1) Results indicated that accumulation of cypermethrin in Carassius auratus Linn, reached balance at the twentieth to twenty fifth days and the maximal enrichment coefficients of muscle and organ were 3.950 and 2.115 . Accumulation capacity of muscle was less than that of liver while cypermethrin release of muscle was quicker than that of liver.
2) There were obvious pathological changes in liver of Carassius auratus Linn, after its continual exposure to cypermethrin for 30 days, but organ coefficients did not occur obvious changes. As for spleen, it did not take on any obvious changes compared with the contrast in anatomization shape and organ coefficients. The influences on lysozyme by cypermethrin in Carassius auratus Linn, were that low concentration lysozymes(2μg·L~(1)-5μg·L~(1)) had the tendency of arising first and declining after, whereas the tendency was contrary for high concentration lysozymes (8μg·L~(1)) . With the increasement of exposure time , all trial groups restored to natural levels at the twentieth day . Lysozemes content in serum was steady until the end of the trial and was not obviously different from the contrast. The outcome revealed that the effects on lysozemes in serum by cypermethrin was reversible for Carassius auratus Linn..
3) After exposure to 1.0ug·L~(-1) of cypermethrin for 30 days , GSH contents and activities of GST、CAT and MAO in muscle and liver had not any significantly difference(P>0.05), but GSH contents and activities of GST、CAT and MAO all occurred dynamic varieties after exposure to cypermethrin above 2.0 ug·L~(-1) : (a) Under the cypermethrin intimidation , the tendency of GSH contents and GST activities were arising first then declining after and the variety degree in liver was bigger than that of muscle . At the end of trial, GSH content in other groups had some changes compared with the contrast except for the group of 1.0 ug·L~(-1), but GST activity was different significantly from the contrast(P<0.01) , 15.71%-67.31% beneath contrast group . (b) CAT activity tendency was decreasing first then arising after and variety degree in liver was bigger than that of muscle. At the end of trial , CAT activity in other groups had some changes compared with the contrast except for the group of 1.0 ug·L~(-1). CAT activity in muscle is lower 17.8% than the contrast group , the most reduction of CAT activity in organ between 51.86%-69.64% (P<0.01); (c) MAO variety was arising first, then declining and restoring at last and variety degree in liver was lower than that in muscle. At the end of trial , MAO activity in other groups had some changes compared with the contrast except for the group of 1.0 ug·L~(-1).
4) The results showed that continual exposure to cypermethrin above 3.0 ug·L~(-1) could induce peripheral red cell to produce micronuclei . Micronuclei frequency had obvious concentration effect and time effect with cypermethrin concentration increased and exposure time prolonged.
The results revealed that cypermethrin above 2.0 ug·L~(-1) produced influences on physiological mechanism of Carassius auratus Linn.; For the security of aquaculture and food available , cypermethrin concentration should be below 1.0 ug·L~(-1) in aquaculture for edible quality.
研究结果表明，氯氰菊酯对平均体长为14．0±1．0cm，平均体重为65．0±5．0g的鲫鱼96hLC_(50)值为18．0μg·L~(-1)，根据农药毒性分级标准，氯氰菊酯对鲫鱼属高毒类农药。
根据急性毒性试验结果，选择1／2 96hLC_(50)以下浓度为试验浓度系列，即1μg·L~(-1)，2μg·L~(-1)，3μg·L~(-1)，5μg·L~(-1)和8μg·L~(-1)对鲫鱼进行连续暴露染毒，分析氯氰菊酯的低剂量连续暴露对鲫鱼的生理生化和遗传指标的影响，结果如下：
1)氯氰菊酯经20-25d可在鲫鱼体内达到蓄积平衡，肌肉和肝脏组织中的最大富集系数分别为3．950和2．155，肌肉组织的蓄积能力小于肝脏组织。在氯氰菊酯的释放过程中，肌肉组织的释放快于肝脏组织。
2)经氯氰菊酯30d连续暴露后鲫鱼肝脏发生了明显的改变，但脏器系数未有显著的变化；而鲫鱼的脾脏无论是解剖形态，还是脏器系数与对照组相比均无明显差异。氯氰菊酯对鲫鱼溶菌酶的影响表现为低浓度组(2μg·L~(-1)-5μg·L~(-1))溶菌酶呈现先升高后降低的趋势，而高浓度组(8μg·L~(-1))则相反，为先降低后升高。但随着暴露时间的延长，各试验组在20-25d时恢复至正常水平，血清中溶菌酶的含量与对照组相比无明显差异，至试验结束时(第30d)各浓度组溶菌酶的含量稳定，变化不大。
3)鲫鱼在1．0μg·L~(-1)氯氰菊酯连续暴露30d后，其肌肉和肝脏中的GSH含量和GST、CAT、MAO的酶活性与对照组相比均未发生显著的差异(P>0．05)，而2．0μg·L~(-1)以上浓度的氯氰菊酯连续暴露对鲫鱼肌肉和肝脏中的GSH含量和GST、CAT、MAO活性均产生了一定程度的影响：(a)氯氰菊酯连续暴露下鲫鱼肌肉和肝脏中的GSH含量和GST活性的变化规律为先升高后降低，且肝脏中的变化要大于肌肉中变化。在试验结束时，除1．0μg·L~(-1)试验组外，其余试验组GSH含量和对照组相比均存在一定的差异；而2．0μg·L~(-1)及以上试验组GST活性在试验结束时与对照组相比差异显著(P<0．01)，低至对照组的15．71％--67．31％。(b)CAT活性的变化规律为先降低后升高，肝脏中的变化幅度要大于肌肉，在试验结束时除1．0μg·L~(-1)试验组外，其余试验组CAT活性与对照组相比均有一定的变化。肌肉中的CAT活性与对照组相比最大降幅可达17．8％，肝脏组织CAT最大降幅可达51．86％-69．64％(P<0．01)。(c) MAO的变化规律为先升高后降低再恢复，在试验结束时除1．0μg·L~(-1)试验组外，其余试验组MAO活性与对照组相比均有一定的变化。
4)3．0μg·L~(-1)以上浓度的氯氰菊酯连续暴露可诱导鲫鱼外周血红细胞产生微核，微核的发生率随暴露浓度的升高和暴露时间的延长具有明显的浓度-效应和时间-效应关系。
研究结果显示，2．0μg·L~(-1)以上浓度的氯氰菊酯可对鲫鱼的生理机能产生一定的影响，这种影响在一定程度上关系到鲫鱼的养殖品质。因此为了鲫鱼的养殖和食用安全，在养殖水环境中氯氰菊酯的浓度应严格控制于1．0μg·L~(-1)以下。
Cypermethrin which was a kind of pyrethroids caused severe contamination to the environment with its wide use in agriculture in recent years . This master paper used Carassius auratus Linn. as experiment animal, applied biochemiscalogy, cell biology etc. technology , studied the influences produced by cypermethrin on Carassius auratus Linn, from acute toxicity trials , accumulation and release rules , immune system , antioxidize system , and heredity effect, from the whole living, organ and cell level and molecule level evaluated the satey of cypermethrin .
The results of acute toxicity revealed that 96hLC_(50) of cypermethrin was 18.0μg·L~(-1) for Carassius auratus Linn. with mean body weight 65.0g±5.0g and mean body length 14.0±1.0cm. According to the methods of biological detect, cypermethrin was a high toxic pesticide for Carassius auratus Linn..
Based on the acute toxicity trial, the following concentration series below 1/2 96hLC_(50) that were 1μg·L~(1),2μg·L~(1),3μg·L~(1) ,5μg/L and 8μg·L~(1) were established to infect Carassius auratus Linn.. Analyzing continual exposure to low dosage cypermethrin for the impact of physiochemical and hereditary of Carassius auratus Linn.. We get a conclusion that:
1) Results indicated that accumulation of cypermethrin in Carassius auratus Linn, reached balance at the twentieth to twenty fifth days and the maximal enrichment coefficients of muscle and organ were 3.950 and 2.115 . Accumulation capacity of muscle was less than that of liver while cypermethrin release of muscle was quicker than that of liver.
2) There were obvious pathological changes in liver of Carassius auratus Linn, after its continual exposure to cypermethrin for 30 days, but organ coefficients did not occur obvious changes. As for spleen, it did not take on any obvious changes compared with the contrast in anatomization shape and organ coefficients. The influences on lysozyme by cypermethrin in Carassius auratus Linn, were that low concentration lysozymes(2μg·L~(1)-5μg·L~(1)) had the tendency of arising first and declining after, whereas the tendency was contrary for high concentration lysozymes (8μg·L~(1)) . With the increasement of exposure time , all trial groups restored to natural levels at the twentieth day . Lysozemes content in serum was steady until the end of the trial and was not obviously different from the contrast. The outcome revealed that the effects on lysozemes in serum by cypermethrin was reversible for Carassius auratus Linn..
3) After exposure to 1.0ug·L~(-1) of cypermethrin for 30 days , GSH contents and activities of GST、CAT and MAO in muscle and liver had not any significantly difference(P>0.05), but GSH contents and activities of GST、CAT and MAO all occurred dynamic varieties after exposure to cypermethrin above 2.0 ug·L~(-1) : (a) Under the cypermethrin intimidation , the tendency of GSH contents and GST activities were arising first then declining after and the variety degree in liver was bigger than that of muscle . At the end of trial, GSH content in other groups had some changes compared with the contrast except for the group of 1.0 ug·L~(-1), but GST activity was different significantly from the contrast(P<0.01) , 15.71%-67.31% beneath contrast group . (b) CAT activity tendency was decreasing first then arising after and variety degree in liver was bigger than that of muscle. At the end of trial , CAT activity in other groups had some changes compared with the contrast except for the group of 1.0 ug·L~(-1). CAT activity in muscle is lower 17.8% than the contrast group , the most reduction of CAT activity in organ between 51.86%-69.64% (P<0.01); (c) MAO variety was arising first, then declining and restoring at last and variety degree in liver was lower than that in muscle. At the end of trial , MAO activity in other groups had some changes compared with the contrast except for the group of 1.0 ug·L~(-1).
4) The results showed that continual exposure to cypermethrin above 3.0 ug·L~(-1) could induce peripheral red cell to produce micronuclei . Micronuclei frequency had obvious concentration effect and time effect with cypermethrin concentration increased and exposure time prolonged.
The results revealed that cypermethrin above 2.0 ug·L~(-1) produced influences on physiological mechanism of Carassius auratus Linn.; For the security of aquaculture and food available , cypermethrin concentration should be below 1.0 ug·L~(-1) in aquaculture for edible quality.
引文
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