摘要
以广东省微生物研究所环境微生物专业菌种库中25个不同种属的菌株为材料,根据菌株与有毒化合物反应的剂量效应关系、系统发育多样性以及对2,3,5-三苯基四氮唑(TTC)响应稳定性的原则筛选了10株细菌和1株酵母菌作为受试菌。结合96孔板建立了一种稳定、灵敏的检测环境污染物急性毒性的微生物新方法,该方法被命名为多种微生物平板毒性测试法(Multispecies Plate for Toxicity testing,MTOXPlate)。MTOXPlate基于11个菌株菌体内脱氢酶活性,活性大小TTC作为细胞活性指示,污染物毒性由脱氢酶抑制率决定。新方法的最佳接种密度(OD600)是1.0稀释100倍,最佳指示剂TTC浓度为0.1%,最佳响应时间是18 h,稳定性较好,重现性相对标准偏差RSD<1%。用本方法测试了7种化合物的急性毒性,并与Microbial Assay for Risk Assessment(MARA)方法作了比较。结果显示:新方法 MTOXPlate与MARA有非常高的相关性(r=0.978,P<0.05),灵敏度略高于MARA方法。
Twenty-five strains were obtained from environmental microbiological culture collection center in Guangdong Microbiology Institute; ten bacteria and one yeast of 25 strains were used as tested materials based on the principles of good dose-response relationship, phylogenetic diversity and stability to 2,3,5-triphenyltetraazolium （TTC）. A new microbiological method was established based on the change of bacteria dehydrogenase activity for monitoring the acute toxicity of chemicals. The dehydrogenase activity was indicated by the reduction rate of TTC. The toxicity of chemicals was calculated according to the inhibition rate of dehydrogenase. The optimum initial inoculation density （OD600） of this method was 1.0, diluted 100 times with LB liquid medium, and the optimum TTC concentration was 0.01%. The mean relative standard deviation （RSD） of microbial toxicity concentration less than 0.1 indicated that the good stability of this method. The toxicity of 7 substances was evaluated with the new method, and the results were compared with those obtained by Microbial Assay for Risk Assessment （MARA）. The data demonstrated that the new method had high correlation with MAMA （r=0.978, P〈0.05） and higher sensitivity slightly than that of MARA. The new method was named as Multispecies Plate for Toxicity testing, MTOXPlate.
作者
关颖
吴泳标
林奕云
张国霞
张胜卿
GUAN Ying WU Yongbiao LIN Yiyun ZHANG Guoxia ZHANG Shengqing(Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China Guangdong Institute of Microbiology, Guangzhou 510070, China Guangzhou Shuguo Bio-Technique Co. Ltd., Guangzhou 510000, China)
出处
《生态科学》
CSCD
2016年第6期6-13,共8页
Ecological Science
基金
国家自然科学基金(31500076)
广东省科技攻关项目(2012B030800011)
广东省科技计划项目(2014A010101135)
南方医科大学公卫学院院长基金(GW201517)
