藻类生长抑制试验中藻细胞计数方法比较  

Comparative Study on Algae Cells Counting Methods of Alga Growth Inhibition Test

作　　者：瞿唯钢[1] 魏方林[2] 刘梅凤[2] 李少南[2] 桂文君[2] 朱国念[2] 

机构地区：[1]中国农业部农药检定所,北京100125 [2]浙江大学农药与环境毒理研究所,杭州310029

出　　处：《农药》2013年第7期497-502,共6页Agrochemicals

摘　　要：[目的]研究农药制剂对藻类生长抑制试验中应用分光光度法、流式细胞仪检测法替代血球计数法用于藻细胞计数的可行性。[方法]采用连续扫描(400-780 nm)确定分光光度法的最佳测试波长;并通过对比血球计数法、分光光度法、流式细胞仪检测法之间的测试结果,为稀释液类型不同的农药选择合适的藻细胞计数方法。[结果]扫描获得对小球藻的最佳检测波长为687 nm;分光光度法与流式细胞仪检测法检测结果间无显著差异,2者与血球计数法所得EC50值有时会存在显著差异,但这种差异不影响毒性等级的划分;稀释液中有微米级固体颗粒存在的农药,采用流式细胞仪检测时易发生堵塞;且流式细胞仪存在将农药稀释液中的固体颗粒与油珠误识别成藻活细胞的可能性。[结论]稀释液无论是哪种类型,分光光度法均可以作为血球计数法的替代方法;对于稀释液为纳米溶液及真溶液的农药,流式细胞仪可以替代血球计数法进行检测。[Aims] The purpose of this study is to apply spectrophotometry and flow cytometric method to replace blood cell count method for counting in alga growth inhibition test for pesticide preparations.[Methods] The best wavelength of spectrophotometry was obtained through continuous scan between 400 and 780 nm.By comparing the test results among blood cell count method,spectrophotometry and flow cytometric method,the appropriate test methods were chosen for pesticides with different type of dilutions.[Results] The best wavelength for spectrophotometry was 687 nm.There was no significant difference between the test results of spectrophotometry and flow cytometric method.Sometimes,values of same pesticides＇ EC50 had significant differences between spectrophotometry or flow cytometric method and blood cell count method,but the differences did not affect the classification of toxicity grade.If using flow cytometric method,clogging may easily occur when micron solid particles abounded in the pesticide dilution,and it was possible to happen that solid particles and oil droplets in pesticide dilution were mistakenly identified as living algae cells.[Conclusions] The results showed that spectrophotometry could replace blood cell count method for algae cells counting in growth inhibition test,and flow cytometric method could also replace blood cell count method for algae cells counting in growth inhibition test when dilution type of pesticide was nanoemulsion or true solution.

关 键 词：农药 藻 生长抑制试验 血球计数法 分光光度法 流式细胞仪检测法 

分 类 号：X173[环境科学与工程—环境科学] S481[农业科学—农药学]