测定环境污染物对青海弧菌发光强度抑制的微板发光法研究

Microplate Luminometry for Toxicity Bioassay of Environmental Pollutant on a New Type of Fresh Water Luminescent Bacterium(Vibrio-qinghaiensis sp.-Q67)

以新型淡水发光菌——青海弧菌Q67(Vibrio-qinghaiensissp.—Q67)为检测生物,以VeritasTM微孔板光度计为发光强度测试设备,建立了测定环境污染物对发光菌发光强度抑制毒性的微板发光测试新方法.系统地研究了pH值、菌密度、反应时间等实验条件对发光强度的影响.应用该方法成功地测定了7种取代酚环境污染物对Q67的发光抑制毒性效应.提出应用非线性迭代最小二乘拟合法模拟环境污染物对Q67毒性的剂量-效应曲线(DRC),拟合效应与实验结果之间的相关系数均大于0.99.通过拟合的DRC参数,准确地计算了污染物的半数效应浓度EC50.对比有关文献方法,微板发光法具有更简便快捷,节省试剂药品,便于多次平行测定从而提高准确度等优点.

A new microplate luminometry for the toxicity bioassay of environmental pollutant on one of fresh water luminescent bacteriums, Vibrio-qinghaiensis sp.-Q67 ,was developed using VeritasTM Microplate Luminometer to measure the luminous intensity of Q67. Effects of pH, bacterial density and reaction time on the bioassay were systematically investigated. The method was successfully employed in the toxic effect test of 7 phenols on Q67. Using non-linear iterative least square technique,the dose-response curves （DRC） of all phenols and heavy metals were accurately fitted with the correlation coefficients between the fitted and observed responses being greater than 0.99. The median effective concentration （ECs0） of all phenols and metals were accurately measured from the DRC models. Compared with some literatures, this bioassay is a fast easy-operate and cost-effective method with high accuracy.