四种除草剂对两种泥鳅的毒性效应
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摘要
本研究以泥鳅和大鳞副泥鳅为受试生物,从两种泥鳅的急性毒性、血清生化指标(GPT、GOT酶活)、DNA损伤水平、胚胎孵化率及畸形率等五个方面综合研究了四种除草剂(草威特、百草枯、氟乐灵、大葱专用除草剂)对两种泥鳅的急性毒性效应、生理毒性效应及遗传毒性效应,为环境污染的监测及合理使用除草剂提供一定的科学依据,并为各类除草剂的毒性研究提供一定的理论基础。
通过急性毒性试验,得出草威特对泥鳅24h、36h、48h的半致死浓度(medium lethal concentration,LC50)分别为0.0736 ml.L~(-1)、0.0685 ml.L~(-1)、0.0661 ml.L~(-1),安全浓度(safe concentration)为0.016 ml.L~(-1)。百草枯对泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.723 ml.L~(-1)、0.660 ml.L~(-1)、0.648 ml.L~(-1),安全浓度(SC)为0.156 ml.L~(-1)。氟乐灵对泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.0697 ml.L~(-1)、0.0654 ml.L~(-1)、0.0625 ml.L~(-1),安全浓度(SC)为0.0151 ml.L~(-1)。大葱专用除草剂对泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.038 ml.L~(-1)、0.027 ml.L~(-1)、0.022 ml.L~(-1),安全浓度(SC)为0.0023 ml.L~(-1)。
草威特对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.0747 ml.L~(-1)、0.0645 ml.L~(-1)、0.0611 ml.L~(-1),安全浓度(SC)为0.0123 ml.L~(-1)。百草枯对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.710 ml.L~(-1)、0.660 ml.L~(-1)、0.614 ml.L~(-1),安全浓度(SC)为0.138 ml.L~(-1)。氟乐灵对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.0672 ml.L~(-1)、0.0613 ml.L~(-1)、0.0591 ml.L~(-1),安全浓度(SC)为0.0137 ml.L~(-1)。大葱专用除草剂对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.032 ml.L~(-1)、0.024 ml.L~(-1)、0.020 ml.L~(-1),安全浓度(SC)为0.0025 ml.L~(-1)。
根据急性毒性试验结果,设置5个浓度组和一个对照组对两种泥鳅进行生理毒性试验,测定两种泥鳅血清中谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性,以判断四种除草剂对两种泥鳅的生理毒性效应。结果表明:四种除草剂均能不同程度上诱使泥鳅和大鳞副泥鳅血浆中谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性的增高,泥鳅和大鳞副泥鳅的肝脏在四种除草剂作用下发生了损伤,四种除草剂对两种泥鳅具有生理毒性作用。但随着四种除草剂处理时间的延长,谷丙转氨酶和谷草转氨酶活性均有所回落。
在急性毒性试验基础上,进一步通过单细胞凝胶电泳技术研究了四种除草剂对两种泥鳅血细胞DNA的损伤,以评价四种除草剂对两种泥鳅的遗传毒性效应。结果表明:草威特、百草枯、大葱专用除草剂对泥鳅和大鳞副泥鳅的血细胞DNA均具有一定程度的损伤作用,对这两种泥鳅均具有遗传毒性作用,但随着这三种除草剂处理时间的延长,部分处理组DNA的损伤程度有所降低,这可能和DNA修复系统的激活有关。氟乐灵对泥鳅血细胞DNA的损伤作用并不明显,对大鳞副泥鳅虽有损伤作用,但这种损伤作用随处理浓度的增加和时间的延长变化并不明显,怀疑是其它因素(如裂解过程及电泳过程中日光的照射等)对血细胞DNA造成了损伤。总的来说,草威特、百草枯、大葱专用除草剂对泥鳅和大鳞副泥鳅具有遗传毒性效应,而氟乐灵则对两种泥鳅的遗传毒性效应并不明显。
在胚胎毒性试验中,以两种泥鳅的孵化率和畸形率为指标,研究四种除草剂对两种泥鳅的胚胎毒性效应,结果发现,草威特、百草枯、氟乐灵、大葱专用除草剂对泥鳅和大鳞副泥鳅胚胎的孵化率均有不同程度的抑制作用,而且具有剂量效应关系。草威特、百草枯、大葱专用除草剂均能不同程度诱发两种泥鳅畸形胚胎的产生,而氟乐灵对两种泥鳅的胚胎并无致畸效应。
We studied the acute toxicity effect,physiological toxicity effect and genetic toxicity effect of four kinds of herbicide on Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage) through a cuople of toxicological methods.Acute toxicity test was applied to determine the LC50 (medium lethal concentration) and SC(safe concentration) of the herbicide on the loach. Biochemical status of two kinds of enzyme relating to liver damage was measured to evalute the physiological toxicity effect.And we further used SCGE(Single Cell Gel Electrophesis) technique to test DNA damage in order to study the toxicity effect of the herbicide generally,which also,we hope,can provide scientific basis for monitoring the contaminating of the herbicide to environment and using herbicide properly.
Through acute toxicity test,the SCs and LC50s of 24h,36h and 48h are acqured.The LC50s of 24h,36h and 48h of quizalofop-p-ethyl on Misgurnus anguillicaudatus are 0.0736 ml.L~(-1),0.0685 ml.L~(-1) and 0.0661 ml.L~(-1) respectively,and the SC of quizalofop-p-ethyl on Misgurnus anguillicaudatus is 0.016 ml.L~(-1). The LC50s of 24h,36h and 48h of Paraquat on Misgurnus anguillicaudatus are 0.723 ml.L~(-1),0.660 ml.L~(-1) and 0.648 ml.L~(-1) respectively,and the SC of Paraquat on Misgurnus anguillicaudatus is 0.156 ml.L~(-1). The LC50s of 24h,36h and 48h of Trifluralin on Misgurnus anguillicaudatus are 0.0697 ml.L~(-1),0.0654 ml.L~(-1) and 0.0625 ml.L~(-1) respectively,and the SC of Trifluralin on Misgurnus anguillicaudatus is 0.0151 ml.L~(-1). The LC50s of 24h,36h and 48h of green onion herbicide on Misgurnus anguillicaudatus are 0.038 ml.L~(-1),0.027 ml.L~(-1) and 0.022 ml.L~(-1) respectively,and the SC of green onion herbicide on Misgurnus anguillicaudatus is 0.0023 ml.L~(-1).
The LC50s of 24h,36h and 48h of quizalofop-p-ethyl on Paramisgurnus dabryanus (Sauvage) are 0.0747 ml.L~(-1),0.0645 ml.L~(-1) and 0.0611 ml.L~(-1) respectively,and the SC of quizalofop-p-ethyl on Paramisgurnus dabryanus (Sauvage) is 0.0123 ml.L~(-1). The LC50s of 24h,36h and 48h of Paraquat on Paramisgurnus dabryanus (Sauvage) are 0.710 ml.L~(-1),0.660 ml.L~(-1) and 0.614 ml.L~(-1) respectively,and the SC of Paraquat on Paramisgurnus dabryanus (Sauvage) is 0.138 ml.L~(-1). The LC50s of 24h,36h and 48h of Trifluralin on Paramisgurnus dabryanus (Sauvage) are 0.0672 ml.L~(-1),0.0613 ml.L~(-1) and 0.0591 ml.L~(-1) respectively,and the SC of Trifluralin on Paramisgurnus dabryanus (Sauvage) is 0.0137 ml.L~(-1). The LC50s of 24h,36h and 48h of green onion herbicide on Misgurnus anguillicaudatus are 0.032 ml.L~(-1),0.024 ml.L~(-1) and 0.020 ml.L~(-1) respectively,and the SC of green onion herbicide on Paramisgurnus dabryanus (Sauvage) is 0.0025 ml.L~(-1).
Based on the results of the acute toxicity test, five experiment groups of different concentration levels and a control group were designed for physiological toxicity test. In the physiological toxicity test,the activity of GPT and GOT was measured.The results showed that four kinds of herbicide all could induce the increse of the GPT and GOT activity in different levels,from which we conclude that the liver of Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage) was damaged by the herbicide and four kinds of herbicide have physiological toxicity effect. But we also found that as time of experiment went on ,the GPT and GOT activity all decresed finally.
To further explore the DNA damage effect of the herbicide we studied, SCGE technique was applied. The results showed that quizalofop-p-ethyl, Paraquat and green onion herbicide all could induce DNA damage of different levels. But as time of experoment went on, the DNA damage levels of several experiment groups decreased. This may be caused by the activating of DNA repair system. The results also showed that Trifluralin had no DNA damage effect on Misgurnus anguillicaudatus. As for Paramisgurnus dabryanus (Sauvage), Trifluralin had little DNA damage effect on it and the DNA damage levels did not change significantly as time and concentration increased,from which we concluded that other factors, for example sunlight,may result this damage. In conclusion, quizalofop-p-ethyl, Paraquat and green onion herbicide have genetic toxicity effect on Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage) and the genetic toxicity effect of Trifluralin need to be further studied.
In the embryo toxicity test,four kinds of herbicide all can reduce the hatching rate of Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage). And quizalofop-p-ethyl, Paraquat and green onion herbicide all can induce malformation embryos and have teratogenic effect on the tested organisms. Trifluralin,however,has no teratogenic effect on Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage).
通过急性毒性试验,得出草威特对泥鳅24h、36h、48h的半致死浓度(medium lethal concentration,LC50)分别为0.0736 ml.L~(-1)、0.0685 ml.L~(-1)、0.0661 ml.L~(-1),安全浓度(safe concentration)为0.016 ml.L~(-1)。百草枯对泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.723 ml.L~(-1)、0.660 ml.L~(-1)、0.648 ml.L~(-1),安全浓度(SC)为0.156 ml.L~(-1)。氟乐灵对泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.0697 ml.L~(-1)、0.0654 ml.L~(-1)、0.0625 ml.L~(-1),安全浓度(SC)为0.0151 ml.L~(-1)。大葱专用除草剂对泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.038 ml.L~(-1)、0.027 ml.L~(-1)、0.022 ml.L~(-1),安全浓度(SC)为0.0023 ml.L~(-1)。
草威特对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.0747 ml.L~(-1)、0.0645 ml.L~(-1)、0.0611 ml.L~(-1),安全浓度(SC)为0.0123 ml.L~(-1)。百草枯对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.710 ml.L~(-1)、0.660 ml.L~(-1)、0.614 ml.L~(-1),安全浓度(SC)为0.138 ml.L~(-1)。氟乐灵对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.0672 ml.L~(-1)、0.0613 ml.L~(-1)、0.0591 ml.L~(-1),安全浓度(SC)为0.0137 ml.L~(-1)。大葱专用除草剂对大鳞副泥鳅24h、36h、48h的半致死浓度(LC50)分别为0.032 ml.L~(-1)、0.024 ml.L~(-1)、0.020 ml.L~(-1),安全浓度(SC)为0.0025 ml.L~(-1)。
根据急性毒性试验结果,设置5个浓度组和一个对照组对两种泥鳅进行生理毒性试验,测定两种泥鳅血清中谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性,以判断四种除草剂对两种泥鳅的生理毒性效应。结果表明:四种除草剂均能不同程度上诱使泥鳅和大鳞副泥鳅血浆中谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性的增高,泥鳅和大鳞副泥鳅的肝脏在四种除草剂作用下发生了损伤,四种除草剂对两种泥鳅具有生理毒性作用。但随着四种除草剂处理时间的延长,谷丙转氨酶和谷草转氨酶活性均有所回落。
在急性毒性试验基础上,进一步通过单细胞凝胶电泳技术研究了四种除草剂对两种泥鳅血细胞DNA的损伤,以评价四种除草剂对两种泥鳅的遗传毒性效应。结果表明:草威特、百草枯、大葱专用除草剂对泥鳅和大鳞副泥鳅的血细胞DNA均具有一定程度的损伤作用,对这两种泥鳅均具有遗传毒性作用,但随着这三种除草剂处理时间的延长,部分处理组DNA的损伤程度有所降低,这可能和DNA修复系统的激活有关。氟乐灵对泥鳅血细胞DNA的损伤作用并不明显,对大鳞副泥鳅虽有损伤作用,但这种损伤作用随处理浓度的增加和时间的延长变化并不明显,怀疑是其它因素(如裂解过程及电泳过程中日光的照射等)对血细胞DNA造成了损伤。总的来说,草威特、百草枯、大葱专用除草剂对泥鳅和大鳞副泥鳅具有遗传毒性效应,而氟乐灵则对两种泥鳅的遗传毒性效应并不明显。
在胚胎毒性试验中,以两种泥鳅的孵化率和畸形率为指标,研究四种除草剂对两种泥鳅的胚胎毒性效应,结果发现,草威特、百草枯、氟乐灵、大葱专用除草剂对泥鳅和大鳞副泥鳅胚胎的孵化率均有不同程度的抑制作用,而且具有剂量效应关系。草威特、百草枯、大葱专用除草剂均能不同程度诱发两种泥鳅畸形胚胎的产生,而氟乐灵对两种泥鳅的胚胎并无致畸效应。
We studied the acute toxicity effect,physiological toxicity effect and genetic toxicity effect of four kinds of herbicide on Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage) through a cuople of toxicological methods.Acute toxicity test was applied to determine the LC50 (medium lethal concentration) and SC(safe concentration) of the herbicide on the loach. Biochemical status of two kinds of enzyme relating to liver damage was measured to evalute the physiological toxicity effect.And we further used SCGE(Single Cell Gel Electrophesis) technique to test DNA damage in order to study the toxicity effect of the herbicide generally,which also,we hope,can provide scientific basis for monitoring the contaminating of the herbicide to environment and using herbicide properly.
Through acute toxicity test,the SCs and LC50s of 24h,36h and 48h are acqured.The LC50s of 24h,36h and 48h of quizalofop-p-ethyl on Misgurnus anguillicaudatus are 0.0736 ml.L~(-1),0.0685 ml.L~(-1) and 0.0661 ml.L~(-1) respectively,and the SC of quizalofop-p-ethyl on Misgurnus anguillicaudatus is 0.016 ml.L~(-1). The LC50s of 24h,36h and 48h of Paraquat on Misgurnus anguillicaudatus are 0.723 ml.L~(-1),0.660 ml.L~(-1) and 0.648 ml.L~(-1) respectively,and the SC of Paraquat on Misgurnus anguillicaudatus is 0.156 ml.L~(-1). The LC50s of 24h,36h and 48h of Trifluralin on Misgurnus anguillicaudatus are 0.0697 ml.L~(-1),0.0654 ml.L~(-1) and 0.0625 ml.L~(-1) respectively,and the SC of Trifluralin on Misgurnus anguillicaudatus is 0.0151 ml.L~(-1). The LC50s of 24h,36h and 48h of green onion herbicide on Misgurnus anguillicaudatus are 0.038 ml.L~(-1),0.027 ml.L~(-1) and 0.022 ml.L~(-1) respectively,and the SC of green onion herbicide on Misgurnus anguillicaudatus is 0.0023 ml.L~(-1).
The LC50s of 24h,36h and 48h of quizalofop-p-ethyl on Paramisgurnus dabryanus (Sauvage) are 0.0747 ml.L~(-1),0.0645 ml.L~(-1) and 0.0611 ml.L~(-1) respectively,and the SC of quizalofop-p-ethyl on Paramisgurnus dabryanus (Sauvage) is 0.0123 ml.L~(-1). The LC50s of 24h,36h and 48h of Paraquat on Paramisgurnus dabryanus (Sauvage) are 0.710 ml.L~(-1),0.660 ml.L~(-1) and 0.614 ml.L~(-1) respectively,and the SC of Paraquat on Paramisgurnus dabryanus (Sauvage) is 0.138 ml.L~(-1). The LC50s of 24h,36h and 48h of Trifluralin on Paramisgurnus dabryanus (Sauvage) are 0.0672 ml.L~(-1),0.0613 ml.L~(-1) and 0.0591 ml.L~(-1) respectively,and the SC of Trifluralin on Paramisgurnus dabryanus (Sauvage) is 0.0137 ml.L~(-1). The LC50s of 24h,36h and 48h of green onion herbicide on Misgurnus anguillicaudatus are 0.032 ml.L~(-1),0.024 ml.L~(-1) and 0.020 ml.L~(-1) respectively,and the SC of green onion herbicide on Paramisgurnus dabryanus (Sauvage) is 0.0025 ml.L~(-1).
Based on the results of the acute toxicity test, five experiment groups of different concentration levels and a control group were designed for physiological toxicity test. In the physiological toxicity test,the activity of GPT and GOT was measured.The results showed that four kinds of herbicide all could induce the increse of the GPT and GOT activity in different levels,from which we conclude that the liver of Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage) was damaged by the herbicide and four kinds of herbicide have physiological toxicity effect. But we also found that as time of experiment went on ,the GPT and GOT activity all decresed finally.
To further explore the DNA damage effect of the herbicide we studied, SCGE technique was applied. The results showed that quizalofop-p-ethyl, Paraquat and green onion herbicide all could induce DNA damage of different levels. But as time of experoment went on, the DNA damage levels of several experiment groups decreased. This may be caused by the activating of DNA repair system. The results also showed that Trifluralin had no DNA damage effect on Misgurnus anguillicaudatus. As for Paramisgurnus dabryanus (Sauvage), Trifluralin had little DNA damage effect on it and the DNA damage levels did not change significantly as time and concentration increased,from which we concluded that other factors, for example sunlight,may result this damage. In conclusion, quizalofop-p-ethyl, Paraquat and green onion herbicide have genetic toxicity effect on Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage) and the genetic toxicity effect of Trifluralin need to be further studied.
In the embryo toxicity test,four kinds of herbicide all can reduce the hatching rate of Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage). And quizalofop-p-ethyl, Paraquat and green onion herbicide all can induce malformation embryos and have teratogenic effect on the tested organisms. Trifluralin,however,has no teratogenic effect on Misgurnus anguillicaudatus and Paramisgurnus dabryanus (Sauvage).
引文
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