氨基甲酸酯类农药对蛋白核小球藻联合毒性作用特点及机制
|
摘要
氨基甲酸酯类农药广泛应用于农业生产中,其在环境中的残留及其对非靶标生物的毒性作用引起关注。以5种氨基甲酸酯类农药包括残杀威、灭多威、抗蚜威、涕灭威和呋喃丹为研究对象,以蛋白核小球藻为受试生物,应用微板毒性分析法系统测定每种农药及其五元混合物对蛋白核小球藻在不同暴露时间(12、24、48、72和96 h)的生长抑制作用,并同步分析农药对蛋白核小球藻的生理特性如叶绿素含量、蛋白质含量、超氧化物歧化酶(SOD)活性和脂质过氧化物丙二醛(MDA)含量的影响。结果表明,5种农药对蛋白核小球藻的浓度-效应均具有明显的时间依赖性,农药抗蚜威在中低浓度促进绿藻生长,呈现非单调J型浓度-效应曲线(CRC)特征,其余4种农药的CRC呈现经典S型;以半数效应浓度的负对数(p EC50)为毒性大小指标,5种农药在96 h时毒性大小为:呋喃丹(p EC50=3.43)>残杀威(p EC50=2.76)>抗蚜威(p EC50=2.12)>灭多威(p EC50=2.11)>涕灭威(p EC50=1.89)。浓度为EC50的5种农药处理后的蛋白核小球藻中叶绿素和蛋白质含量随暴露时间的延长而减少,但不同农药处理的绿藻中叶绿素和蛋白质含量减少率随暴露时间延长变化趋势稍有不同; SOD酶活性随着暴露时间延长逐渐下降,MDA含量逐渐增加,这说明藻细胞受到破坏,脂质过氧化的损害程度超过细胞修复能力,SOD活性被抑制,细胞的抗氧化能力下降,藻细胞内的H2O2不断积累,导致MDA含量升高。五元混合物对蛋白核小球藻的毒性也具有一定的时间依赖性,并表现出刺激作用,即hormesis现象,且混合物毒性与组分浓度比具有良好的线性相关性;暴露于混合物的小球藻均在96 h出现了刺激效应,其叶绿素与蛋白质含量随暴露时间延长不断增加,SOD活性不断升高,MDA含量不断减少;五元混合物均在96 h呈现出拮抗作用,且与混合物的浓度和组分浓度比相关。
Carbamate pesticides are widely used in agricultural production,so their residues in the environment and toxicity towards non-target organisms are concerned. Five kinds of carbamate pesticides,i. e. baygon,pirimicarb,methomyl,aldicarb and carbofuran were selected as research substances,the microplate toxicity analysis method was used to determine the growth inhibition toxicity of each pesticide and their mixtures towards Chlorella pyrenoidsa in different exposure times(0,24,48,72 and 96 h). The effects of the five pesticides and their mixtures on the physiological characeristics such as chlorophyll content,protein content,superoxide dismutase(SOD) activity and lipid peroxidation product malondialdehyde(MDA) content in Chlorella pyrenoidosa were also investigated to explore possible toxicity mechanisms. The results showed that the five pesticides had time-dependent effects. The pesticide pirimicarb promoted the growth of green algae at low and medium concentrations,showing a non-monotonic Jtype concentration-response curve(CRC). The CRCs of the other four pesticides showed classic S types. Selecting the negative logarithm of the half effect concentration(pEC50) as the toxicity index,toxicity order of the five pesticides in 96 h was as follows: carbofuran(p EC50= 3.43) > baygon(p EC50= 2.77) > pirimicarb(p EC50= 2.12) >methomyl(p EC50= 2.11) > aldicarb(pEC50= 1.89). The chlorophyll and protein contents as well as SOD activity in Chlorella pyrenoidosa decreased gradually with the lengthening of exposure time. However,MDA contents increased with time lengthening,showing that the algae cells were destructed and the lipid peroxidation exceeded the self-repairing ability of the cells. This caused the inhibition of SOD activity,and thus harmed the antioxidant capacity of cells. Thus,the accumulation of H2 O2 in algae cells resulted in the decreasing of SOD activity and increasing of MDA content. Toxicity of the five pesticide mixtures was also time-dependent and showed stimulatory effect,i.e.hormesis phenomenon. The maximum stimulating effects of the five-component mixtures appeared in 96 h,and the chlorophyll and protein content together with SOD activity also increased. As a result,the content of MDA was continuously decreased. Antagonism appeared in the five-component mixtures in 96 h,which was closely related to the concentrations or concentration ratios of mixture components.
Carbamate pesticides are widely used in agricultural production,so their residues in the environment and toxicity towards non-target organisms are concerned. Five kinds of carbamate pesticides,i. e. baygon,pirimicarb,methomyl,aldicarb and carbofuran were selected as research substances,the microplate toxicity analysis method was used to determine the growth inhibition toxicity of each pesticide and their mixtures towards Chlorella pyrenoidsa in different exposure times(0,24,48,72 and 96 h). The effects of the five pesticides and their mixtures on the physiological characeristics such as chlorophyll content,protein content,superoxide dismutase(SOD) activity and lipid peroxidation product malondialdehyde(MDA) content in Chlorella pyrenoidosa were also investigated to explore possible toxicity mechanisms. The results showed that the five pesticides had time-dependent effects. The pesticide pirimicarb promoted the growth of green algae at low and medium concentrations,showing a non-monotonic Jtype concentration-response curve(CRC). The CRCs of the other four pesticides showed classic S types. Selecting the negative logarithm of the half effect concentration(pEC50) as the toxicity index,toxicity order of the five pesticides in 96 h was as follows: carbofuran(p EC50= 3.43) > baygon(p EC50= 2.77) > pirimicarb(p EC50= 2.12) >methomyl(p EC50= 2.11) > aldicarb(pEC50= 1.89). The chlorophyll and protein contents as well as SOD activity in Chlorella pyrenoidosa decreased gradually with the lengthening of exposure time. However,MDA contents increased with time lengthening,showing that the algae cells were destructed and the lipid peroxidation exceeded the self-repairing ability of the cells. This caused the inhibition of SOD activity,and thus harmed the antioxidant capacity of cells. Thus,the accumulation of H2 O2 in algae cells resulted in the decreasing of SOD activity and increasing of MDA content. Toxicity of the five pesticide mixtures was also time-dependent and showed stimulatory effect,i.e.hormesis phenomenon. The maximum stimulating effects of the five-component mixtures appeared in 96 h,and the chlorophyll and protein content together with SOD activity also increased. As a result,the content of MDA was continuously decreased. Antagonism appeared in the five-component mixtures in 96 h,which was closely related to the concentrations or concentration ratios of mixture components.
引文
[1]华小梅，单正军．我国农药的生产，使用状况及其污染环境因子分析[J]．环境科学进展，1996,4(2):33-45Hua X M,Shan Z J. The production and application of pesticides and factor analysis of their pollution in environment in China[J]. Advances in Environmental Science,1996,4(2):33-45(in Chinese)
[2]潘媛媛，刘晓达，金燕，等．柱后衍生法分析8种氨基甲酸酯农药[J]．食品安全导刊，2015(25):30-32
[3]王未，黄从建，张满成，等．我国区域性水体农药污染现状研究分析[J]．环境保护科学，2013,39(5):5-9Wang W,Huang C J,Zhang M C,et al. Study on status of regional water pollution by pesticides in China[J].Environmental Protection Science,2013,39(5):5-9(in Chinese)
[4]孙英，张立金，闵顺耕，等．三种氨基甲酸酯类农药化合物对DNA的潜在损伤作用[J]．农业环境科学学报，2004,23(3):464-466Sun Y,Zhang L J,Min S G,et al. The potential trauma of three kinds of carbamate pesticides to DNA[J]. Journal of Agro-Environment Science,2004,23(3):464-466(in Chinese)
[5]张瑾，董欣琪，陈敏，等．五元氨基甲酸酯类农药混合物体系对青海弧菌的毒性特点[J]．生态毒理学报，2017,12(4):138-145Zhang J,Dong X Q,Chen M,et al. Toxicity characterstics of five-carbamate pesticide mixture system towards Vibrio qinghaiensis sp. Q67[J]. Asian Journal of Ecotoxicology,2017,12(4):138-145(in Chinese)
[6]夏更寿．氨基甲酸酯类农药对水华鱼腥藻的毒性效应[J]．安徽农业科学，2005,33(3):391-392Xia G S. Toxicity effect of six carbamic ester pesticides on Anabaena flos-aquae[J]. Journal of Anhui Agricultural Sciences,2005,33(3):391-392(in Chinese)
[7] Wang Q Q,Lemley A T. Competitive degradation and detoxification of carbamate insecticides by membrane anodic fenton treatment[J]. Journal of Agricultural and Food Chemistry,2003,51(18):5382-5390
[8]余涛，陈秀杰，谭倩，等．饮用水中8种有机磷类农药和除草剂及氨基甲酸酯类农药的超高效液相色谱-串联质谱直接测定法[J]．环境与健康杂志，2015,32(11):1014-1016Yu T,Chen X J,Tan Q,et al. Simultaneous determination of eight kinds of organophosphorus pesticides,urethane pesticides and herbicide in drinking water by direct injection ultra-performance liquid chromatography tandem mass spectrometry[J]. Journal of Environment and Health,2015,32(11):1014-1016(in Chinese)
[9]沈国兴，严国安．农药对藻类的生态毒理学研究:II．毒性机理及其富集和降解[J]．环境科学进展，1999,7(6):131-140Shen G X,Yan G A. Study on ecotoxicology for pesticides to algae II:Toxic mechanism and accumulation,degradation[J]. Advances in Environmental Science,1999,7(6):131-140(in Chinese)
[10] El-Sheekh M M,Kotkat H M,Hammouda O H E.Effect of atrazine herbicide on growth,photosynthesis,protein synthesis,and fatty acid composition in the unicellular green alga Chlorella kessleri[J]. Ecotoxicology&Environmental Safety,1994,29(3):349-358
[11]刘永涛，李乐，杨红，等．3种渔用药物对斜生栅藻的毒性效应研究[J]．生态环境学报，2017,26(2):261-267Liu Y T,Li L,Yang H,et al. Toxic effects of three fishery drugs on Scenedesmus obliquus[J]. Ecology and Environment Sciences,2017,26(2):261-267(in Chinese)
[12]姜慧，丁婷婷，张瑾，等．3种有机溶剂及其混合物对蛋白核小球藻时间毒性的探究[J]．安徽农业大学学报，2018,45(3):480-486Jiang H,Ding T T,Zhang J,et al. The time-dependent toxicity of three organic solvents and their mixtures towards Chlorella pyrenoidosa[J]. Journal of Anhui Agricultural University,2018,45(3):480-486(in Chinese)
[13]陈敏，张瑾，董欣琪，等．多元抗生素与重金属混合物对蛋白核小球藻的时间依赖性协同与拮抗作用[J]．农业环境科学学报，2018,37(5):850-859Chen M,Zhang J,Dong X Q,et al. Time-dependent synergism and antagonism within multi-component mixtures of heavy metals and antibiotics towards Chlorella pyrenoidosa[J]. Journal of Agro-Environment Science,2018,37(5):850-859(in Chinese)
[14]徐冬梅，王艳花，饶桂维．四环素类抗生素对淡水绿藻的毒性作用[J]．环境科学，2013,34(9):3386-3390Xu D M,Wang Y H,Rao G W. Cellular response of freshwater green algae to the toxicity of tetracycline antibiotics[J]. Environmental Science,2013,34(9):3386-3390(in Chinese)
[15]袁静，刘树深，王丽娟，等．蛋白核小球藻(Chlorella pyrenoidosa)微板毒性分析方法优化[J]．环境科学研究，2011,24(5):553-558Yuan J,Liu S S,Wang L J,et al. Optimization of microplate toxicity analysis method based on Chlorella pyrenoidose[J]. Research of Environmental Sciences,2011,24(5):553-558(in Chinese)
[16]肖菊，刘树深，高音旋，等．均匀设计用于研究硝基苯衍生物对青海弧菌Q67的联合毒性[J]．环境科学研究，2008,21(2):120-124Xiao J,Liu S S,Gao Y X,et al. Use of uniform design to evaluate the joint toxicity of nitrobenzene derivatives to Vibrio qinghaiensis sp.-Q67[J]. Research of Environmental Sciences,2008,21(2):120-124(in Chinese)
[17] Liu S S,Xiao Q F,Zhang J,et al. Uniform design ray in the assessment of combined toxicities of multi-component mixtures[J]. Science Bulletin,2016,61(1):52-58
[18]袁静，刘树深，王丽娟．10种咪唑离子液体藻微板毒性分析[J]．环境科学与技术，2010,33(12F):46-49Yuan J,Liu S S,Wang L J. Microplate toxicity analysis of 10 imidazolium ionic liquid to Chlorella pyrenonidosa[J]. Environmental Science&Technology,2010,33(12F):46-49(in Chinese)
[19]陈琼，张瑾，李小猛，等．几种抗生素对蛋白核小球藻的时间毒性微板分析法[J]．生态毒理学报，2015,10(2):190-197Chen Q,Zhang J,Li X M,et al. Time-dependent microplate toxicity analysis(T-MTA)of several antibiotics to Chlorella pyrenoidosa[J]. Asian Journal of Ecotoxicology,2015,10(2):190-197(in Chinese)
[20] Ge H L,Liu S S,Zhu X W,et al. Predicting hormetic effects of ionic liquid mixtures on luciferase activity using the concentration addition model[J]. Environmental Science&Technology,2011,45(4):1623-1629
[21] Qin L T,Liu S S,Zhang J,et al. A novel model integrated concentration addition with independent action for the prediction of toxicity of multi-component mixture[J]. Toxicology,2011,280(3):164-172
[22]刘树深，刘玲，陈浮．浓度加和模型在化学混合物毒性评估中的应用[J]．化学学报，2013,71(10):1335-1340Liu S S,Liu L,Chen F. Application of the concentration addition model in the assessment of chemical mixture toxicity[J]. Acta Chimica Sinica,2013,71(10):1335-1340(in Chinese)
[23] Liu S S,Song X Q,Liu H L,et al. Combined photobacterium toxicity of herbicide mixtures containing one insecticide[J]. Chemosphere,2009,75(3):381-388
[24] Zhang Y H,Liu S S,Song X Q,et al. Prediction for the mixture toxicity of six organophosphorus pesticides to the luminescent bacterium Q67[J]. Ecotoxicology and Environmental Safety,2008,71(3):880-888
[25]卓静，荆国华，李小林，等．表面活性剂CTAC和STAB对蛋白核小球藻的毒作用[J]．农业环境科学学报，2010,29(8):1460-1465Zhuo J,Jing G H,Li X L,et al. Toxic effects of surfactants CTAC and STAB on Chlorella pyrenoidosa[J].Journal of Agro-Environment Science,2010,29(8):1460-1465(in Chinese)
[26]陈彦，朱奇，张永忠．植物超越补偿作用的研究进展[J]．自然杂志，2000,22(2):88-91Chen Y,Zhu Q,Zhang Y Z. Overcompensation responses of plants:An overview[J]. Ziran Zazhi,2000,22(2):88-91(in Chinese)
[27]李瑷伶，蓝昕，司万童，等．己烯雌酚对染镉草鱼GSH-Px、SOD和MDA影响的研究[J]．农业环境科学学报，2008,27(1):350-353Li A L,Lan X,Si W T,et al. Effects of diethylstilbestrol on GSH-Px,SOD and MDA of grass carp exposed to cadmium[J]. Journal of Agro-Environment Science,2008,27(1):350-353(in Chinese)
[2]潘媛媛，刘晓达，金燕，等．柱后衍生法分析8种氨基甲酸酯农药[J]．食品安全导刊，2015(25):30-32
[3]王未，黄从建，张满成，等．我国区域性水体农药污染现状研究分析[J]．环境保护科学，2013,39(5):5-9Wang W,Huang C J,Zhang M C,et al. Study on status of regional water pollution by pesticides in China[J].Environmental Protection Science,2013,39(5):5-9(in Chinese)
[4]孙英，张立金，闵顺耕，等．三种氨基甲酸酯类农药化合物对DNA的潜在损伤作用[J]．农业环境科学学报，2004,23(3):464-466Sun Y,Zhang L J,Min S G,et al. The potential trauma of three kinds of carbamate pesticides to DNA[J]. Journal of Agro-Environment Science,2004,23(3):464-466(in Chinese)
[5]张瑾，董欣琪，陈敏，等．五元氨基甲酸酯类农药混合物体系对青海弧菌的毒性特点[J]．生态毒理学报，2017,12(4):138-145Zhang J,Dong X Q,Chen M,et al. Toxicity characterstics of five-carbamate pesticide mixture system towards Vibrio qinghaiensis sp. Q67[J]. Asian Journal of Ecotoxicology,2017,12(4):138-145(in Chinese)
[6]夏更寿．氨基甲酸酯类农药对水华鱼腥藻的毒性效应[J]．安徽农业科学，2005,33(3):391-392Xia G S. Toxicity effect of six carbamic ester pesticides on Anabaena flos-aquae[J]. Journal of Anhui Agricultural Sciences,2005,33(3):391-392(in Chinese)
[7] Wang Q Q,Lemley A T. Competitive degradation and detoxification of carbamate insecticides by membrane anodic fenton treatment[J]. Journal of Agricultural and Food Chemistry,2003,51(18):5382-5390
[8]余涛，陈秀杰，谭倩，等．饮用水中8种有机磷类农药和除草剂及氨基甲酸酯类农药的超高效液相色谱-串联质谱直接测定法[J]．环境与健康杂志，2015,32(11):1014-1016Yu T,Chen X J,Tan Q,et al. Simultaneous determination of eight kinds of organophosphorus pesticides,urethane pesticides and herbicide in drinking water by direct injection ultra-performance liquid chromatography tandem mass spectrometry[J]. Journal of Environment and Health,2015,32(11):1014-1016(in Chinese)
[9]沈国兴，严国安．农药对藻类的生态毒理学研究:II．毒性机理及其富集和降解[J]．环境科学进展，1999,7(6):131-140Shen G X,Yan G A. Study on ecotoxicology for pesticides to algae II:Toxic mechanism and accumulation,degradation[J]. Advances in Environmental Science,1999,7(6):131-140(in Chinese)
[10] El-Sheekh M M,Kotkat H M,Hammouda O H E.Effect of atrazine herbicide on growth,photosynthesis,protein synthesis,and fatty acid composition in the unicellular green alga Chlorella kessleri[J]. Ecotoxicology&Environmental Safety,1994,29(3):349-358
[11]刘永涛，李乐，杨红，等．3种渔用药物对斜生栅藻的毒性效应研究[J]．生态环境学报，2017,26(2):261-267Liu Y T,Li L,Yang H,et al. Toxic effects of three fishery drugs on Scenedesmus obliquus[J]. Ecology and Environment Sciences,2017,26(2):261-267(in Chinese)
[12]姜慧，丁婷婷，张瑾，等．3种有机溶剂及其混合物对蛋白核小球藻时间毒性的探究[J]．安徽农业大学学报，2018,45(3):480-486Jiang H,Ding T T,Zhang J,et al. The time-dependent toxicity of three organic solvents and their mixtures towards Chlorella pyrenoidosa[J]. Journal of Anhui Agricultural University,2018,45(3):480-486(in Chinese)
[13]陈敏，张瑾，董欣琪，等．多元抗生素与重金属混合物对蛋白核小球藻的时间依赖性协同与拮抗作用[J]．农业环境科学学报，2018,37(5):850-859Chen M,Zhang J,Dong X Q,et al. Time-dependent synergism and antagonism within multi-component mixtures of heavy metals and antibiotics towards Chlorella pyrenoidosa[J]. Journal of Agro-Environment Science,2018,37(5):850-859(in Chinese)
[14]徐冬梅，王艳花，饶桂维．四环素类抗生素对淡水绿藻的毒性作用[J]．环境科学，2013,34(9):3386-3390Xu D M,Wang Y H,Rao G W. Cellular response of freshwater green algae to the toxicity of tetracycline antibiotics[J]. Environmental Science,2013,34(9):3386-3390(in Chinese)
[15]袁静，刘树深，王丽娟，等．蛋白核小球藻(Chlorella pyrenoidosa)微板毒性分析方法优化[J]．环境科学研究，2011,24(5):553-558Yuan J,Liu S S,Wang L J,et al. Optimization of microplate toxicity analysis method based on Chlorella pyrenoidose[J]. Research of Environmental Sciences,2011,24(5):553-558(in Chinese)
[16]肖菊，刘树深，高音旋，等．均匀设计用于研究硝基苯衍生物对青海弧菌Q67的联合毒性[J]．环境科学研究，2008,21(2):120-124Xiao J,Liu S S,Gao Y X,et al. Use of uniform design to evaluate the joint toxicity of nitrobenzene derivatives to Vibrio qinghaiensis sp.-Q67[J]. Research of Environmental Sciences,2008,21(2):120-124(in Chinese)
[17] Liu S S,Xiao Q F,Zhang J,et al. Uniform design ray in the assessment of combined toxicities of multi-component mixtures[J]. Science Bulletin,2016,61(1):52-58
[18]袁静，刘树深，王丽娟．10种咪唑离子液体藻微板毒性分析[J]．环境科学与技术，2010,33(12F):46-49Yuan J,Liu S S,Wang L J. Microplate toxicity analysis of 10 imidazolium ionic liquid to Chlorella pyrenonidosa[J]. Environmental Science&Technology,2010,33(12F):46-49(in Chinese)
[19]陈琼，张瑾，李小猛，等．几种抗生素对蛋白核小球藻的时间毒性微板分析法[J]．生态毒理学报，2015,10(2):190-197Chen Q,Zhang J,Li X M,et al. Time-dependent microplate toxicity analysis(T-MTA)of several antibiotics to Chlorella pyrenoidosa[J]. Asian Journal of Ecotoxicology,2015,10(2):190-197(in Chinese)
[20] Ge H L,Liu S S,Zhu X W,et al. Predicting hormetic effects of ionic liquid mixtures on luciferase activity using the concentration addition model[J]. Environmental Science&Technology,2011,45(4):1623-1629
[21] Qin L T,Liu S S,Zhang J,et al. A novel model integrated concentration addition with independent action for the prediction of toxicity of multi-component mixture[J]. Toxicology,2011,280(3):164-172
[22]刘树深，刘玲，陈浮．浓度加和模型在化学混合物毒性评估中的应用[J]．化学学报，2013,71(10):1335-1340Liu S S,Liu L,Chen F. Application of the concentration addition model in the assessment of chemical mixture toxicity[J]. Acta Chimica Sinica,2013,71(10):1335-1340(in Chinese)
[23] Liu S S,Song X Q,Liu H L,et al. Combined photobacterium toxicity of herbicide mixtures containing one insecticide[J]. Chemosphere,2009,75(3):381-388
[24] Zhang Y H,Liu S S,Song X Q,et al. Prediction for the mixture toxicity of six organophosphorus pesticides to the luminescent bacterium Q67[J]. Ecotoxicology and Environmental Safety,2008,71(3):880-888
[25]卓静，荆国华，李小林，等．表面活性剂CTAC和STAB对蛋白核小球藻的毒作用[J]．农业环境科学学报，2010,29(8):1460-1465Zhuo J,Jing G H,Li X L,et al. Toxic effects of surfactants CTAC and STAB on Chlorella pyrenoidosa[J].Journal of Agro-Environment Science,2010,29(8):1460-1465(in Chinese)
[26]陈彦，朱奇，张永忠．植物超越补偿作用的研究进展[J]．自然杂志，2000,22(2):88-91Chen Y,Zhu Q,Zhang Y Z. Overcompensation responses of plants:An overview[J]. Ziran Zazhi,2000,22(2):88-91(in Chinese)
[27]李瑷伶，蓝昕，司万童，等．己烯雌酚对染镉草鱼GSH-Px、SOD和MDA影响的研究[J]．农业环境科学学报，2008,27(1):350-353Li A L,Lan X,Si W T,et al. Effects of diethylstilbestrol on GSH-Px,SOD and MDA of grass carp exposed to cadmium[J]. Journal of Agro-Environment Science,2008,27(1):350-353(in Chinese)