费氏弧菌急性毒性微孔板检测方法优化

Optimization of acute toxicity test method using microplate for Vibrio fischeri

为优化费氏弧菌(Vibrio fischeri)急性毒性微孔板法测试条件,以发光强度或发光抑制率为指标,研究温度、盐度、p H、反应体系、稀释比例、传代次数对费氏弧菌新鲜菌液生物毒性测试的影响。结果表明:费氏弧菌适宜培养温度为15~20℃;检测适宜温度为15℃,盐度为15‰~35‰,p H为5~10,检测体系为1∶10,传代次数不宜超过10代,不同稀释比例对新鲜培养的生长平台期费氏弧菌毒性测试无显著影响。基于优化后的方法,七水硫酸锌、氯化汞、十二烷基硫酸钠、苯酚对费氏弧菌急性毒性具有显著的剂量-效应关系,表明该优化方法适用于上述物质的毒性评价。

In order to determine the optimal conditions of acute toxicity test method using microplate for luminescent bacteria（ Vibrio fischeri）,the effects of temperature,salinity,pH,detection temperature,testing system,dilution ratio,passages on the bioassay of V. fischeri were systematically studied with Luminous intensity or inhibitory effect on luminescent bacteria. The results showed that the growth curve of V. fischeri reached plateau and peak value faster at 20℃,but there was no significant difference of growth density and peak value at 15℃ and20℃. The appropriate culture temperature of V. fischeri was 15℃-20℃;The optimal test temperature,salinity,pH value and test system were 15℃,15‰-35‰,5-10 and 1∶10,respectively. The bioassay results were significantly affected by both test temperature and test system of V. fischeri. There was no significant difference of bioassay results in different dilution ratio（ the range of initial luminescence intensity from 10 w to 600 w）,but the acute toxicity test results of V. fischeri were significantly affected after 15 times of passages. The toxicity of ZnSO4·7 H2 O,HgCl2,SDS,phenol for V. fischeri had time responses and dose responses by the method above. It showed that the acute toxicity test method using microplate for V. fischeri was suitable for the toxicity assessment of pollutants above.