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

LIU Bao-qi,GE Hui-lin,LIU Shu-shen,
刘保奇
,葛会林,刘树深

生态毒理学报 , 2006,
Abstract: 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 (EC50) 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.
Detection of autoinducer in Vibrio qinghaiensis sp. -Q67 and its effect on luminescence
青海弧菌Q67自体诱导物的检测及其对菌体发光的影响

YE Jiang-Yu,CHEN Yu,LI Shu-Yue,ZHAI Jun,HE Qiang,SHU Wei-Qun,
叶姜瑜
,陈宇,李书钺,翟俊,何强,舒为群

微生物学通报 , 2011,
Abstract: Vibrio qinghaiensis sp. -Q67 was a newly identified luminescent bacterium in fresh water, its character of luminous intensity changed with toxic pollutant concentrations made it as an strain for water quality monitoring. Through bioassay strain JZA1 test, C18 reverse phase thin-layer chromatography and LacZ activity test, it was identified that Q67 had a LuxI-LuxR type Quorum-Sensing and produced N-acyl-homoserine lactone (AHL) as autoinducer. The other test results indicated that the content and activity of the signal molecular in Q67 changed with its growth phases. The crude extract from Q67 affected not only its luminescence but also its growth and reproduction.
Characterizing the Toxicity Interaction of the Binary Mixture Between DMSO and Pesticide by the Multi-Effect Residual Analysis (MERA)
多效应残差法(MERA)表征二甲亚砜-农药二元混合物毒性相互作用

HUO Xiang-chen,LIU Shu-shen,ZHANG Jing,ZHANG Jin,
霍向晨
,刘树深,张晶,张瑾

环境科学 , 2013,
Abstract: Three groups of binary mixtures between dimethylsulfoxide (DMSO) and three widely used pesticides, dimethoate (DIM), dichlorvos (DIC), and metalaxyl (MET), were respectively constructed by using the direct equipartition ray design (EquRay). The luminescent inhibition toxicities of single chemical and binary mixtures to Vibrio qinghaiensis sp.-Q67 were determined by the microplate toxicity analysis (MTA). Selecting the concentration addition (CA) model as an additive reference, we developed a new multi-effect residual analysis (MERA) to quantitatively characterize the deviation of the observed toxicity from that predicted by the CA model, i. e. the degree of toxicity interaction. It was shown that the toxicity interactions between DMSO and pesticide were dominated by antagonism, and the highest antagonism distributed between -23% and -15%. The concentration ranges where antagonism existed and the degree of antagonism were influenced by the components in the mixture, the concentration ratios of the components, and the effect level. A comparison of the MERA with the conventional isobologram and the extended toxic unit summation revealed that the MERA characterizes the degree of toxicity interaction in the view of effect, with less limitation by different concentration ratios or effect levels. Therefore, the MERA can be used to evaluate the complex toxicity interactions taking place in binary mixtures.
Combined toxicity of ionic liquids and industrial wastewater on Vibrio qinghaiensis sp.-Q67
离子液体与废水对青海弧菌Q67的混合毒性研究

ZHANG Jin,LIU Shushen,WANG Chenglin,DENG Huiping,
张瑾
,刘树深,王成林,邓慧萍

环境科学学报 , 2013,
Abstract: By selecting two ionic liquids (ILs), 1-benzyl-3-methylimidazolium tetrafluoroborate (IL1) and 1-hexyl-3-methyl-imidazolium bis (trifluoromethylsulfony)imide (IL2), and wastewater (WW) as mixture components, two mixture systems, one between IL1 and IL2 and the other among IL1, IL2 and WW with different concentration ratios (pis), were designed by the uniform design ray procedure. The toxicities of single components and the mixtures to Vibrio qinghaiensis sp.-Q67 (Q67) were determined by the microplate toxicity analysis. The toxicity interaction within mixtures was analyzed by using the concentration addition and independent action as reference models. It was showed that the mixtures between IL1 and IL2 exhibited antagonistic or additive interaction and the combined toxicity (pEC50) was well correlated with the concentration percentage of IL1, while the mixtures among IL1, IL2 and WW displayed synergistic or additive interaction and the combined toxicity (pEC50) was well correlated with the concentration percentage of WW.
Quantitative Structure-Activity Relationship for Toxicity of Fifteen Substituted Phenols to Vibrio qinghaiensis sp.-Q67
15种取代酚对淡水发光菌Q67的毒性及定量构效分析

Zhang Hui,Li N,Ma Mei,Liu Guangbin,
张辉
,李娜,马梅,刘光斌

生态毒理学报 , 2012,
Abstract: 为了更加准确和便捷地预测各种取代酚类化合物的急性毒性,以淡水发光菌Q67(Vibrio qinghaiensis sp.-Q67)为受试生物,测定了15种典型取代酚的急性毒性;采用logD(正辛醇/水分配系数),LUMO(分子最低空轨道能)和MW(分子量)等取代酚的7种主要结构参数,利用偏最小二乘回归法建立了定量结构-活性相关(quantitative structure-activity relationships,QSAR)模型.结果表明,15种取代酚的EC50在5.76×10-6 ~ 1.27×10-3 mol·L-1之间,且有很好的剂量-效应关系;QSAR模型的主成分分析显示,-logEC50与logD、LUMO和MW值正相关,且logD对模型的贡献最大,即越容易与Q67菌结合的酚类化合物对其的急性毒性越大;建立的QSAR模型具有较好的预测能力(Q2EXT=0.91,RMSE=0.49)和较高的稳定性(Q2CUM=0.58),能够用于预测其他酚类化合物对Q67菌的急性毒性.
Cytotoxicity and genotoxicity evaluation of urban surface waters using freshwater luminescent bacteria Vibrio-qinghaiensis sp.-Q67 and Vicia faba root tip
Xiaoyan M,Xiaochang Wang,Yongjun Liu,
Xiaoyan M
,Xiaochang Wang,Yongjun Liu

环境科学学报(英文版) , 2012,
Abstract: The freshwater luminescent bacteria Vibrio-qinghaiensis sp.-Q67 test and the Vicia faba root tip test associated with solid-phase extraction were applied for cytotoxicity and genotoxicity assessment of organic substances in three rivers, two lakes and effluent flows from two wastewater treatment plants (WWTPs) in Xi'an, China. Although the most seriously polluted river with high chemical oxygen demand (COD) and total organic carbon (TOC) showed high cytotoxicity (expressed as TII50, the toxicity impact index) and genotoxicity (expressed as RMCN, the relative frequency of micronucleus), no correlative relation was found between the ecotoxicity and organic content of the water samples. However, there was a linear correlative relation between TII50 and RMCN for most water samples except that from the Zaohe River, which receives discharge from WWTP and untreated industrial wastewaters. The ecotoxicity of the organic toxicants in the Chanhe River and Zaohe River indicated that cytotoxic and genotoxic effects were related to the pollutant source. The TII50 and RMCN were also found to correlate roughly to the dissolved oxygen concentration of the water. Sufficient dissolved oxygen in surface water is thus proved to be an indicator of a healthy water environmental condition.
Toxicities of Selected Ionic Liquids and Their Mixtures to Photobacteria (Vibrio-qinghaiensis sp. -Q67)
部分离子液体及其混合物对发光菌的毒性作用

LIU Fang,LIU Shu-shen,LIU Hai-ling,
刘芳
,刘树深,刘海玲

生态毒理学报 , 2007,
Abstract: Ionic liquids(ILs)are claimed to be environmentally safe and very good non-volatile solvents for a wide range of applications. Although the information about physical, thermodynamic, kinetic or engineering data has been extended continuously, only little data with regard to the toxicity and ecotoxicity of ILs have been available until now. Using the microplate luminometer as testing equipment and the freshwater photobacteria--Vibrio-qinghaiensis sp.-Q67 as indication organism, the luminescence inhibition toxicities of 12 ionic liquids including C6H11BF4N2(S1), C8H15ClN2(S2), C8H15BF4N2(S3), C9H14BF4N(S4), C9H17BF4N2(S5), C9H17BrN2(S6), C11H13BF4N2(S7), C11H13ClN2(S8), C12H23BrN2(S9), C14H27BF4N2(S10), C14H27ClN2(S11), and C16H31ClN2(S12) and their mixtures to Q67 were determined. Obviously, there were 2 groups of mixtures, a high-toxicity group(H-set) with pEC50 > 4.5, including S9, S10, S11 and S12, and a low-toxicity group (L-set) with pEC50 < 3.5, including S1, S2, S3, S4, S5, S6, S7 and S8. The EC50 values of single compounds(from S1 to S12)revealed that the toxic order was S10>S12>S11>S9>S7>S8>S6>S3>S4>S2>S5>S1. The test mixtures were prepared using equivalent-effect concentration ratio(EECR)and uniform design concentration ratio(UDCR)method. It has been found that the toxicities of 8 test mixtures designed by the EECR and the H-set or L-set ILs can be accurately predicted by the dose-addition(DA)model. Also, the toxicities of 6 test mixtures constructed by the UDCR and the H-set ILs can be predicted by the DA model. However, the toxicities of 6 test mixtures constructed by the UDCR and the L-set ILs can't be effectively predicted by the DA or independent action(IA)model because there is an upwarp in the low-dose area of the dose-response curves.
Toxicities of Selected Heavy Metal Compounds and Their Mixtures to Photobacteria( Vibrio-qinghaiensis sp. —Q67)
部分重金属化合物对淡水发光菌的毒性研究

DENG Fu-cai,LIU Shu-shen,LIU Hai-ling,MO Ling-yun,
邓辅财
,刘树深,刘海玲,莫凌云

生态毒理学报 , 2007,
Abstract: 应用微板毒性分析方法,分别测定了CdCl2·2.5H2O、CoSO4·5H2O、Cr(NO3)3·3H2O、Cu(NO3)2·3H2O、Fe(NO3)3·3H2O、MnCl2·9H2O、Na2SeO3、ZnSO4·7H2O、Ni(NO3)2·6H2O 9种重金属离子化合物及其混合物对淡水发光菌-青海弧菌Q67(Vibrio-qinghaiensis sp.-Q67)的发光抑制毒性.结果表明,9种重金属离子化合物对Q67的剂量-效应关系均可用Weibull或Logit模型有效描述.由拟合剂量-效应曲线得到这9种重金属离子化合物的半数效应浓度EC50的负对数值(-logEC50)分别为4.35、3.08、2.39、3.83、3.34、2.39、3.32、3.93和2.76,说明其毒性顺序为:CdCl2·2.5H2O>ZnSO4·7H2O>Cu(NO3)2·3H2O>Fe(NO3)3·3H2O>Na2SeO3>CoSO4·5H2O>Ni(NO3)2·6H2O>Cr(NO3)3·3H2O≈MnCl2·9H2O.为了研究重金属混合物的毒性规律,设计了4组等效应浓度(EC50、EC15、EC10和EC5)比混合物,测试了其混合物毒性,并应用剂量加和(DA)、独立作用(IA)原理及经典联合毒性评价方法进行了分析.DA与IA分析表明,所研究的4种混合物的毒性具有拮抗特征,而毒性单位法(TU)和混合指数法(MTI)的评价结果均为部分相加作用,相加指数法(AI)的评价结果则为拮抗作用.所选评价方法不同,混合物毒性评价结果可能也不同.
Hormesis response of marine and freshwater luminescent bacteria to metal exposure
SHEN,KAILI; SHEN,CHAOFENG; LU,YUAN; TANG,XIANJIN; ZHANG,CONGKAI; CHEN,XINCAI; SHI,JIYAN; LIN,QI; CHEN,YINGXU;
Biological Research , 2009, DOI: 10.4067/S0716-97602009000200006
Abstract: the stimulatory effect of low concentrations of toxic chemicals on organismal metabolism, referred to as hormesis, has been found to be common in the widely used luminescence bioassay. this paper aims to study the hormesis phenomenon in both marine and freshwater luminescent bacteria, named photobacterium phosphorem and vibrio qinghaiensis. the effects of cu (ii), zn (ii), cd (ii) and cr (vi) on luminescence of these two bacteria were studied for 0 to 75 minutes exposure by establishing dose- and time-response curves. a clear hormesis phenomenon was observed in all four testing metals at low concentrations under the condition of luminescence assays.
Hormesis response of marine and freshwater luminescent bacteria to metal exposure
KAILI SHEN,CHAOFENG SHEN,YUAN LU,XIANJIN TANG
Biological Research , 2009,
Abstract: The stimulatory effect of low concentrations of toxic chemicals on organismal metabolism, referred to as hormesis, has been found to be common in the widely used luminescence bioassay. This paper aims to study the hormesis phenomenon in both marine and freshwater luminescent bacteria, named Photobacterium phosphorem and Vibrio qinghaiensis. The effects of Cu (II), Zn (II), Cd (II) and Cr (VI) on luminescence of these two bacteria were studied for 0 to 75 minutes exposure by establishing dose- and time-response curves. A clear hormesis phenomenon was observed in all four testing metals at low concentrations under the condition of luminescence assays.
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