Gold immunochromatographic assay for simultaneous detection of carbofuran and triazophos in water samples

Using a simple test for rapid identification and quantification of pesticide multiresidues in food and environmental samples is a long-cherished approach for practical monitoring purposes. Here two gold-based lateral-flow strips (strip A and strip B) were investigated for simultaneous detection of carbofuran and triazophos. For the strip A format, a bispecific monoclonal antibody (BsMcAb) against both carbofuran and triazophos was employed to prepare the immunogold probe. For the strip B format, anti-carbofuran monoclonal antibody (McAb) and anti-triazophos McAb separately labeled with colloidal gold were combined as detector reagents. By comparison of visual results from pesticide standard tests between the two formats, the strip B assay manifested higher sensitivities for both pesticides. Analysis of spiked water samples by the preferable strip indicated that the detection limits for carbofuran and triazophos were 32 and 4 μg/L, respectively. The strength of the portable one-step strip assay was in the simultaneous screening for two pesticides within a short time (8-10 min) without any equipment.     

三唑磷降解菌的筛选及其降解途径研究

从三唑磷生产厂周围的土壤中用土壤富集的方法筛选分离出一株三唑磷降解菌Klebsiella sp.,它能以三唑磷为唯一碳源、唯一氮源、唯一磷源生长同时实现对三唑磷的降解,三唑磷作为唯一氮源时的降解速度最快,是实现三唑磷降解的最佳营养方案。在三唑磷为唯一氮源时,1000 mg/L的三唑磷浓度对菌体降解无抑制作用。此降解菌首先通过水解作用实现对TAP 的降解,之后把水解产物进一步降解为无毒的无机物质。降解菌的这些降解特性表明了它用于生物降解消除三唑磷污染的巨大潜力。     

三种农药对斑马鱼的急性毒性和生物浓缩系数

采用斑马鱼(Brachydanio rerio)为试验生物,研究了吡虫啉、三唑磷和哒螨灵的急性毒性和生物浓缩系数．通过毒性试验,获得了这3种农药对斑马鱼的24、48、72和96h LC50值．这3种农药的96h LC50值分别为281．37mg·L^-1、8．37mg·L^-1和11．66μg·L^-1．根据急性毒性数据和有关技术指南,确定生物浓缩试验浓度和持续暴露时间为14d．经0．5和5mg·L^-1吡虫啉暴露后的生物浓缩系数(BCFs)分别为1．52和0．97．经0．02和0．2mg·L^-1三唑磷暴露后的BCFs分别为9．00和8．45．经0．1和1μg·L^-1哒螨灵暴露后的BCFs分别为5600和4920．     

Mutation in acetylcholinesterase1 associated with triazophos resistance in rice stem borer, Chilo suppressalis (Lepidoptera: Pyralidae)

Two full-length genes encoding different acetylcholinesterases (AChEs), designated as Ch-ace1 and Ch-ace2, were cloned from strains of the rice stem borer (Chilo suppressalis) susceptible and resistant to the organophosphate insecticide triazophos. Sequence analysis found an amino acid mutation A314S in Ch-ace1 (corresponding to A201 in Torpedo californica AChE) that was consistently associated with the occurrence of resistance. This mutation removed an MspA1 I restriction site from the wild type allele. An assay based on restriction fragment length polymorphism (RFLP) analysis was developed to diagnose A314S genotypes in field populations. Results showed a strong correlation between frequencies of the mutation and phenotypic levels of resistance to triazophos. The assay offers a prospect for rapid monitoring of resistance and assisting with the appropriate choice of insecticide for combating damage caused by C. suppressalis.     

三唑磷降解菌株GS-1的分离鉴定及其降解特性的研究

从有机磷农药污水处理池污泥中分离到一株能高效降解三唑磷的菌株GS-1, 通过生理生化实验和16S rDNA序列同源性分析, 将该菌株鉴定为Diaphorobacter sp.。菌株GS-1能以三唑磷为唯一碳源生长, 能在12 h内降解100 mg/L的三唑磷至检测不出的水平。菌株GS-1降解三唑磷的过程中会产生中间代谢产物苯唑醇(1-苯基-3-羟基-1,2,4-三唑), 36 h后苯唑醇被完全转化。菌株GS-1降解三唑磷的最适pH值为8.0, 最适温度为30°C, 且对杀螟硫磷、辛硫磷、毒死蜱和甲基对硫磷     

Hapten synthesis for enzyme-linked immunoassay of the insecticide triazophos

Two haptens of the insecticide triazophos (O,O-diethyl O-[1-phenyl-1H-1,2,4-triazol-3-yl] phosphorothioate) were synthesized by introducing appropriate spacers in the O-ethyl site of the analyte molecular structure. First, thiophosphoryl chloride (PSCl(3)) reacts with methanol at low temperature to give O-ethyl dichlorothiophosphate. After reacting with 1-phenyl-3H-1,2,4-triazol, the O-ethyl dichlorothiophosphate was transformed into the intermediate O-ethyl O-(1-phenyl-1H-1,2,4-triazol-3-yl) chlorothiophosphate. Then the intermediate reacts with 4-aminobutyric acid and 6-aminobutyric acid to produce hapten I and hapten II, respectively. The molecule structures of the two haptens were identified by (1)H nuclear magnetic resonance spectrum and mass spectrum. An enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibody was also developed to evaluate the two haptens. Results showed that the monoclonal antibodies with high titers were obtained after immunizing with protein conjugates of these haptens and that the immunoassay has high affinity and specificity to triazophos. These results suggested that the haptens were synthesized successfully and could be used for immunoassay for the rapid screening and sensitive determination of this insecticide.     

三唑磷、氟虫腈及其复配剂对四种miRNAs在斑马鱼组织中表达的影响

MicroRNAs(miRNAs)是一类长约22 nt的非编码小RNA,在基因表达中起重要调控作用.已有研究表明,农药三唑磷和氟虫腈能影响斑马鱼全鱼组织中部分miRNAs的正常表达,但未见对miRNA表达的组织特异性的研究.本研究采用荧光定量PCR技术研究了经三唑磷微乳剂、氟虫腈微乳剂及其复配剂处理后,四种miRNA(...     

Development of a one-step strip for the detection of triazophos residues in environmental samples

Environmental and food safety issues now are recognized internationally, and pesticide residues play key roles as environment and food pollutants. It is crucial to develop methods for rapid determination of pesticide residues in environments and foods. A one-step strip based on nanocolloidal-gold-labeled monoclonal antibodies for detection of triazophos residue was developed. The nanocolloidal gold, with an average particle diameter of 25 nm (G25), was labeled to an antitriazophos monoclonal antibody. This conjugate was dispensed on the conjugate pad of a porous glass fiber. Ovalbumin hapten and goat anti-mouse IgG were dispensed on the nitrocellulose membrane and served as the test line (T-line) and control line (C-line), respectively. After conditions optimization, the one-step strip was finally developed for the residue determination of triazophos. The limit of detection (LOD) of the strip was 4 ng/mL for standard. The detection was not affected by the pH of the liquid sample but low total ion concentration will induce illegible C-line and T-line. The LOD for spiked samples of soil and water was 5 ng/mL, with run time of no more than 10 min.