基于叶绿素荧光成像技术的藻毒性检测法的建立及在环境监测中的应用
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摘要
应用新型大面积调制叶绿素荧光成像系统(Maxi-Imaging-PAM),以蛋白核小球藻作为指示生物,以典型的光合作用抑制剂敌草隆和莠灭净为测试物质,建立了微孔板式藻毒性检测方法,确定了实验参数对结果的影响,并将该方法应用于检测石油废水的藻毒性.结果表明,最适实验条件为:藻细胞的暴露浓度在3.45×10~6～5.28×10~6 cell·mL~(-1)范围内,暴露时间为2 h时,毒性检测最灵敏,获得的敌草隆和莠灭净的半最大效应浓度(EC_(50))分别为(4.33±0.38)μg·L~(-1)和(3.87±0.27)μg·L~(-1).将该方法成功应用于某石油废水的藻毒性检测,确定其EC_(50)值为1.03%±0.07%(稀释百分数),相当于414.96μg·L~(-1)的敌草隆的毒性.本研究建立的微孔板式藻毒性检测方法具有灵敏度高、所需样品量少、可大批量测试等优点,对毒理学发展具有重要意义.
Using a novel large-area modulating chlorophyll fluorescence imaging system(Maxi-Imaging-PAM), a method for algae toxicity test was developed and optimized based on microplate. In this method, Chlorella pyrenoidosa was the testing organism, and typical photosynthesis inhibitors diuron and ametryn were chosen as testing substances. The influence of the experimental parameters on the results were discussed in this study. Finally, this developed method was applied on the algae toxicity test of petroleum wastewater. The results showed that when the concentration of the algal solution was 3.45×10~6～5.28×10~6 cells·mL~(-1) and exposure time was 2 hours, the method displayed a good testing property. At this condition, the concentration of 50% maximal effect(EC_(50)) of diuron and ametryn were(4.33±0.38) μg·L~(-1) and(3.87±0.27) μg·L~(-1), respectively. The method was successfully applied to the algae toxicity test of petroleum wastewater, and its EC_(50) value was 1.03%±0.07%(dilution percentage), which is equivalent to 414.96 μg·L~(-1) diuron. The established microplate algae toxicity detection method has the advantages of high sensitivity, less sample volume demanded, and large batch test. This has great significance for the development of toxicology.
Using a novel large-area modulating chlorophyll fluorescence imaging system(Maxi-Imaging-PAM), a method for algae toxicity test was developed and optimized based on microplate. In this method, Chlorella pyrenoidosa was the testing organism, and typical photosynthesis inhibitors diuron and ametryn were chosen as testing substances. The influence of the experimental parameters on the results were discussed in this study. Finally, this developed method was applied on the algae toxicity test of petroleum wastewater. The results showed that when the concentration of the algal solution was 3.45×10~6～5.28×10~6 cells·mL~(-1) and exposure time was 2 hours, the method displayed a good testing property. At this condition, the concentration of 50% maximal effect(EC_(50)) of diuron and ametryn were(4.33±0.38) μg·L~(-1) and(3.87±0.27) μg·L~(-1), respectively. The method was successfully applied to the algae toxicity test of petroleum wastewater, and its EC_(50) value was 1.03%±0.07%(dilution percentage), which is equivalent to 414.96 μg·L~(-1) diuron. The established microplate algae toxicity detection method has the advantages of high sensitivity, less sample volume demanded, and large batch test. This has great significance for the development of toxicology.
引文
Alexandre Camuel A,Guieysse B,Cynthia Alcántara,et al.2017.Fast algal eco-toxicity assessment: Influence of light intensity and exposure time on Chlorella vulgaris inhibition by atrazine and DCMU[J].Ecotoxicology & Environmental Safety,140:141
Cedergreen N,Kudsk P,Mathiassen S K,et al.2007.Combination effects of herbicides on plants and algae: do species and test systems matter[J].Pest Management Science,63(3):282-295
丛佳.2017.污水处理厂二级出水回用于景观用水的生物安全性评价[D].天津:天津理工大学
杜丽娜,曹宇,穆玉峰,等.2014.羊角月牙藻在制药废水毒性评价中的应用[J].环境科学研究,27(12):1525-1531
Fait G,Balderacchi M,Ferrari F,et al.2010.A field study of the impact of different irrigation practices on herbicide leaching[J].European Journal of Agronomy,32(4):280-287
Focazio M J,Kolpin D W,Barnes K K,et al.2008.A national reconnaissance for pharmaceuticals and other organic wastewater contaminants in the United States — II) Untreated drinking water sources[J].Science of the Total Environment,402(2):201-216
Forsberg C G.1972.Algal assay procedure[J].Journal,44(8):1623-1628
葛会林,刘树深,苏冰霞.2014.通用浓度加和模型预测有机磷与三嗪农药对绿藻的联合毒性[J].中国环境科学,34(9): 2413-2419
黄飞羽,段招财,林彦伶,等.2017.饮用水的细菌及藻类检测与卫生调查[J].基层医学论坛,21(25):3437-3438
Hasenbein S,Peralta J,Lawler S P,et al.2017.Environmentally relevant concentrations of herbicides impact non-target species at multiple sublethal endpoints[J].Science of the Total Environment,607-608:733-743
Hillebrand O,N?dler K,Geyer T,et al.2014.Investigating the dynamics of two herbicides at a karst spring in Germany: Consequences for sustainable raw water management[J].Science of the Total Environment,482-483(3):193-200
刘威.2015.阿特拉津对地表饮用水源水质影响评价及毒理研究[D].长春:东北师范大学
Ma J,Lin F,Wang S,et al.2003.Toxicity of 21 herbicides to the green alga Scenedesmus quadricauda[J].Bulletin of Environmental Contamination & Toxicology,71(3):594
Ma J,Xu L,Wang S,et al.2002.Toxicity of 40 herbicides to the green alga Chlorella vulgaris[J].Ecotoxicology & Environmental Safety,51(2):128
Meyer M T,Thurman E M.2012.Herbicide metabolites in surface water and groundwater[J].Organic Letters,14(16):4035-4037
Muller R,Schreiber U,Escher B I,et al.2008.Rapid exposure assessment of PSII herbicides in surface water using a novel chlorophyll a,fluorescence imaging assay[J].Science of the Total Environment,401(1):51-59
钱海丰,陈思,金瑜剑.2017.藻类在除草剂生物毒性安全评估中的应用[J].浙江工业大学学报,45(1):32-36
Schreiber U,Müller J F,Haugg A,et al.2002.New type of dual-channel PAM chlorophyll fluorometer for highly sensitive water toxicity biotests[J].Photosynthesis Research,74(3):317-330
Sjollema S B,van Beusekom S A,Hg V D G,et al.2014.Laboratory algal bioassays using PAM fluorometry: Effects of test conditions on the determination of herbicide and field sample toxicity[J].Environmental Toxicology & Chemistry,33(5):1017-1022
史媛,王英才,李晔,等.2013.基于藻类荧光的毒性测试研究进展[J].环境科学与技术,13(3):59-64
王丽,应波,鄂学礼.2008.水中敌草隆的叶绿素荧光检测法[J].环境与健康杂志,25(6):539-541
严登华,何岩,王浩.2005.东辽河流域地表水体中Atrazine的环境特征[J].环境科学,26(3):203-208
叶丹,陈洁,袁琳,等.2014.除草剂对3种绿藻的毒性测试研究[J].人民长江,2014(18):82-86
虞龙,苗秀珍,于洋,等.2010.应用新型调制荧光成像系统(Imaging-PAM)原位评估底泥毒性的方法[J].环境化学,29(6):1116-1120
Cedergreen N,Kudsk P,Mathiassen S K,et al.2007.Combination effects of herbicides on plants and algae: do species and test systems matter[J].Pest Management Science,63(3):282-295
丛佳.2017.污水处理厂二级出水回用于景观用水的生物安全性评价[D].天津:天津理工大学
杜丽娜,曹宇,穆玉峰,等.2014.羊角月牙藻在制药废水毒性评价中的应用[J].环境科学研究,27(12):1525-1531
Fait G,Balderacchi M,Ferrari F,et al.2010.A field study of the impact of different irrigation practices on herbicide leaching[J].European Journal of Agronomy,32(4):280-287
Focazio M J,Kolpin D W,Barnes K K,et al.2008.A national reconnaissance for pharmaceuticals and other organic wastewater contaminants in the United States — II) Untreated drinking water sources[J].Science of the Total Environment,402(2):201-216
Forsberg C G.1972.Algal assay procedure[J].Journal,44(8):1623-1628
葛会林,刘树深,苏冰霞.2014.通用浓度加和模型预测有机磷与三嗪农药对绿藻的联合毒性[J].中国环境科学,34(9): 2413-2419
黄飞羽,段招财,林彦伶,等.2017.饮用水的细菌及藻类检测与卫生调查[J].基层医学论坛,21(25):3437-3438
Hasenbein S,Peralta J,Lawler S P,et al.2017.Environmentally relevant concentrations of herbicides impact non-target species at multiple sublethal endpoints[J].Science of the Total Environment,607-608:733-743
Hillebrand O,N?dler K,Geyer T,et al.2014.Investigating the dynamics of two herbicides at a karst spring in Germany: Consequences for sustainable raw water management[J].Science of the Total Environment,482-483(3):193-200
刘威.2015.阿特拉津对地表饮用水源水质影响评价及毒理研究[D].长春:东北师范大学
Ma J,Lin F,Wang S,et al.2003.Toxicity of 21 herbicides to the green alga Scenedesmus quadricauda[J].Bulletin of Environmental Contamination & Toxicology,71(3):594
Ma J,Xu L,Wang S,et al.2002.Toxicity of 40 herbicides to the green alga Chlorella vulgaris[J].Ecotoxicology & Environmental Safety,51(2):128
Meyer M T,Thurman E M.2012.Herbicide metabolites in surface water and groundwater[J].Organic Letters,14(16):4035-4037
Muller R,Schreiber U,Escher B I,et al.2008.Rapid exposure assessment of PSII herbicides in surface water using a novel chlorophyll a,fluorescence imaging assay[J].Science of the Total Environment,401(1):51-59
钱海丰,陈思,金瑜剑.2017.藻类在除草剂生物毒性安全评估中的应用[J].浙江工业大学学报,45(1):32-36
Schreiber U,Müller J F,Haugg A,et al.2002.New type of dual-channel PAM chlorophyll fluorometer for highly sensitive water toxicity biotests[J].Photosynthesis Research,74(3):317-330
Sjollema S B,van Beusekom S A,Hg V D G,et al.2014.Laboratory algal bioassays using PAM fluorometry: Effects of test conditions on the determination of herbicide and field sample toxicity[J].Environmental Toxicology & Chemistry,33(5):1017-1022
史媛,王英才,李晔,等.2013.基于藻类荧光的毒性测试研究进展[J].环境科学与技术,13(3):59-64
王丽,应波,鄂学礼.2008.水中敌草隆的叶绿素荧光检测法[J].环境与健康杂志,25(6):539-541
严登华,何岩,王浩.2005.东辽河流域地表水体中Atrazine的环境特征[J].环境科学,26(3):203-208
叶丹,陈洁,袁琳,等.2014.除草剂对3种绿藻的毒性测试研究[J].人民长江,2014(18):82-86
虞龙,苗秀珍,于洋,等.2010.应用新型调制荧光成像系统(Imaging-PAM)原位评估底泥毒性的方法[J].环境化学,29(6):1116-1120
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