降解乙酰甲胺磷农药的芽孢杆菌分离鉴定与降解性能分析

[目的]探究微生物对农药乙酰甲胺磷的生物降解作用。[方法]利用筛选培养基从土壤中分离乙酰甲胺磷降解性菌株,对其中一株降解性能较好的菌株YJ1进行分析,扩增菌株16S r DNA,测序并构建系统演化树,结合生理生化性质,初步确定其分类地位,并检测该菌株对乙酰甲胺磷的降解性能。[结果]乙酰甲胺磷降解菌株YJ1属于芽孢杆菌属,可以在乙酰甲胺磷为唯一碳源的环境中存活;该菌株在乙酰甲胺磷含量为1 200 mg/L的培养基中生长良好;在p H7~10碱性条件下,该菌株在乙酰甲胺磷液体培养基中生长较好,酸性条件不利于其生长;该菌株在72 h对乙酰甲胺磷降解率可达100%,具有很高的乙酰甲胺磷降解活性。[结论]所发现的芽孢杆菌YJ1可用于乙酰甲胺磷的生物降解。     

黑土和棕壤中甲胺磷的根际降解脱毒模拟研究

利用塑料根际盒研究了甲胺磷在黑土和棕壤大豆根际和非根际环境中的降解脱毒行为.结果表明,甲胺磷虽是急性毒性较高的农药,但在土壤环境中能很快降解,并且同等条件下,甲胺磷在黑土中的残留量普遍低于棕壤.在无大豆种植情况下(对照处理),培养试验第2天,棕壤甲胺磷残留量约为33%,黑土只有26%.在大豆根际圈中,甲胺磷的降解明显加快,尤其是在黑土中.第9天,根际盒中层黑土和棕壤的农药残留分别比无植物对照低87.5%和76.0%.甲胺磷的土壤降解过程符合一级动力学方程,降解半衰期为2 d左右.     

细菌表面展示技术在有机磷农药降解中的应用

通过微生物代谢作用来生物降解有机磷农药被认为是安全有效的途径,而近年来发展的细胞表面展示技术,由于是将农药降解酶定位在细胞表面,具有提高反应效率、可再生和减少对细胞的毒害等多种优点,从而为有机磷农药的生物降解与转化提供了一种新的技术对策,已显示出广阔的应用前景。     

甲胺磷降解细菌的分离鉴定及其降解效能的研究

从土壤中分离筛选获得两株对甲胺磷农药有较强降解效能的细菌,经鉴定分别为头状葡萄球菌(Staphylococcus capitis)(称为D菌)和粪产碱菌(Alcaligenes faecalis)(称为J菌)。D菌和J菌在甲胺磷浓度为500mg.L-1,30℃,180r.min-1摇床上用基础培养基中培养72h,对甲胺磷的降解率分别达到58.49%和65.80%。D菌和J菌混合培养可提高对甲胺磷的降解效能,对甲胺磷72h的降解率达到72.93%。     

利用海洋微生物降解有机磷农药MAP的研究

目的分离及筛选降解海水养殖区甲胺磷的降解菌,并确定最适的降解条件。方法从被有机磷污染的海水样中分离,以有机磷为唯一碳源反复驯化,分离筛选出1株高效降解甲胺磷的菌株M-1,并对其降解能力和所需条件进行测试。通过离子交换层析、凝胶过滤层析等方法从发酵液中分离纯化了有机磷农药降解酶。结果初步鉴定菌株M-1属于腊样芽胞杆菌。菌株M-1最适生长温度和pH分别为25℃和8．0。Zn^2＋（200mg／L）、Cd^2＋（50mg／L）与Pb^2＋（200mg／L）不影响菌株M-1对甲胺磷的降解作用,但Cu^2＋（50mg／L）、Cr^2＋（50mg／L）对菌株M-1有毒性作用。SDS-PAGE测得降解菌的有机磷农药降解酶的分子质量约为45kD。结论海洋微生物在甲胺磷污染的海水养殖区自净中起着重要作用。     

定量结构-生物降解性能关系模型在21世纪农药发展中的应用

简要概述农药的概念、环保型农药的主要特点、农药的主要生产和设计途径以及农药在粮食增产的同时给环境带来的负面影响。定量结构一生物降解性能关系（QSBR）能够定量预测有机物的生物降解性能,可作为一种预测有机物降解过程和性能的模型。本文提出将其应用于环保型农药的设计和筛选中,从而减少由于农药生物降解给环境带来的影响。     

甲胺磷降解菌的筛选及降解特性研究

从长期受有机磷农药污染的土壤中分离到1株能降解甲胺磷的菌株B15,经生理生化鉴定为巨大芽孢杆菌(Bacillus megaterium)。在甲胺磷无机盐培养基(甲胺磷浓度为0.5%)生长时,最适生长温度为28℃,最适pH为7.0,摇床培养(28℃190 r/min)48 h降解率达到83%。菌株在甲胺磷浓度为1%的无机盐培养基上能生长,但是在甲胺磷浓度为0.5%的无机盐培养基上生长最好,降解率最高。外加碳氮源对菌株的降解率有所提高,但是超过某一浓度降解率随着浓度的增加反而下降。     

拟除虫菊酯类农药降解菌及降解酶的研究概况

拟除虫菊酯类农药目前已成为我国出口的蔬菜、水果中主要的3类农药残留之一,引起急慢性中毒事件也越来越多,对人类、水生生物和自然环境造成很大危险.而农药生物降解作为去除农药污染的有效手段,逐渐成为环境科学研究的热点.重点综述了拟除虫菊酯类农药降解茵的分离、降解酶的提取、纯化、降解机理、固定化研究,并对以后要解决的问题进行了展望.     

拟除虫菊酯类农药降解菌及降解酶的研究概况

拟除虫菊酯类农药目前已成为我国出口的蔬菜、水果中主要的3类农药残留之一, 引起急慢性中毒事件也越来越多, 对人类、水生生物和自然环境造成很大危险。而农药生物降解作为去除农药污染的有效手段, 逐渐成为环境科学研究的热点。重点综述了拟除虫菊酯类农药降解菌的分离、降解酶的提取、纯化、降解机理、固定化研究, 并对以后要解决的问题进行了展望。     

餐厨废弃物资源化利用的微生物技术研究进展

简单介绍餐厨废弃物的特征和危害,综述微生物技术处理餐厨废弃物资源化的途径,如发酵提取生物降解塑料技术、厌氧发酵处理技术、微生物堆肥技术、微生物农药技术、微生物产电技术,介绍利用复合微生物菌剂降解餐厨废弃物的研究进展,分析这一新技术的发展趋势。     

甲胺磷对拟环纹豹蛛中肠蛋白消化酶活性影响的压电传感监测（英文）

利用压电体声波阻抗分析法实时动态监测了拟环纹豹蛛Pardosa pseudoannulata中肠蛋白消化酶对酪蛋白的酶促水解过程及不同浓度梯度（0.008%、0.016%、0.024%和0.032%）的甲胺磷农药对酶活性的影响,并用紫外分光光度法进行了验证。结果表明,合适低剂量(0.008%)的甲胺磷农药可显著地增强拟环纹豹蛛蛋白消化酶的活性,较高剂量(0.032%)的农药却显著抑制蛋白消化酶的活性,且水解过程稳态频移值可准确地反映酶活性与农药浓度的关系。本工作为研究低剂量农药增强蜘蛛控虫力的机理及酶活性的快速检测提供了一种可行的新方法。     

Positive correlation of methamidophos resistance between Lipaphis erysimi and Diaeretilla rapae and effects of methamidophos ingested by host insect on the parasitoid

Temporal and spatial correlated variations on the methamidophos resistance and biomolecular rate constant of acetylcholinesterase to insecticides were found between the turnip aphid, Lipaphis erysimi and its endoparasitoid, Diaeretilla rapae , collected from field colonies and an insecticide-free field insectarium in Fujian, China. Compared to the related susceptible insectarium population, L. erysimi and D. rapae displayed 7.4–29.2- and 2.6–9.2-fold resistance ratios, respectively. In addition, two populations of L. erysimi with different methamidophos resistance levels, that is, a field (with 5.8-fold resistance ratio) and an insectarium population, were used to study the effects of methamidophos ingested by the host insect on D. rapae development. The percentage of D. rapae cocoon formation decreased significantly when the parasitized L. erysimi were fed on cauliflower leaves treated with methamidophos at lethal concentration dosages 10 (LC₁₀) or LC₅₀. At LC₅₀ dosages the percentage of D. rapae cocoon formation and adult emergence decreased significantly. When the parasitized L. erysimi were fed on methamidophos at LC₉₀ dosage, no D. rapae cocoons were found. When the field or insectarium L. erysimi were treated with methamidophos at LC₁₀, the susceptibility to methamidophos in the adult D. rapae emerged from the treated host insect was similar to the control. However, the susceptibility to methamidophos in the adult D. rapae became lower than the control when the host insects were treated at LC₅₀ dosages. The data thus suggested that the methamidophos ingested by the host insect L. erysimi could be an important factor in the endoparasitoids' insecticide resistance development. The natural selectivity would favor the parasitoids that had developed an insensitivity to the insecticide(s).     

假单胞菌S-2降解甲胺磷性能的研究

从甲胺磷生产车间分离到一株假单胞菌编号为S-2。S-2可利用甲胺磷为唯一氮源,但不能利用甲胺磷为唯一磷源。该文对S-2体内具有的降解甲胺磷的酶类进行了研究,初步断定：S-2可代谢产生酸性磷酸酶,主要在胞外降解甲胺磷。S-2在甲胺磷诱导的情况下,这些降解酶类可大量聚积。用诱导过的菌液降解甲胺磷比未经诱导的快了2d左右。     

Effect of methamidophos on the growth rate and esterase activity of the common carp Cyprinus carpio L.

The acute toxicity of methamidophos and three other compounds to fingerlings of the common carp Cyprinus carpio L. was determined. The 96-h LC₅₀ values were 68 mg/litre for methamidophos, 1.7 mg/litre for carbaryl, 0.21 mg/litre for lindane and 50 mg/litre for diquat.The acetylcholinesterase (AChE) and carboxylesterase (CarE) activities of the brain and liver were monitored over six weeks in fish poisoned with sublethal doses of methamidophos. Brain CarE was more sensitive than AChE but for the liver the reverse was true. In either case the degree of enzyme inhibition increased with increasing insecticide concentrations in the water.After exposure to methamidophos at 20 mg/litre for 48h liver AChE and CarE recovered faster than those of the brain. For both organs CarE recovered faster than AChE.At sublethal doses methamidophos affected the growth rate of the fish but no direct relationship between growth and insecticide concentrations could be established.     

不同地理种群小菜蛾对甲胺磷的抗药性

采用浸叶法研究了福州建新镇、福建农林大学校园内、闽侯县上街镇和闽侯县甘蔗镇等地田间小菜蛾种群对甲胺磷的抗性水平及抗性机制。结果表明,与敏感小菜蛾相比,上述4个田间小菜蛾种群已对甲胺磷产生了34-82倍的抗性水平。敏感小菜蛾乙酰胆碱酯酶(AChE)的Ki值是田间抗性小菜蛾AChE的Ki值的17.3-39.1倍。不同小菜蛾种群对甲胺磷的抗性水平也存在着差异,小菜蛾种群AChE对甲胺磷的敏感性大小与其对甲胺磷的抗性水平高低显著相关。     

Selective solid-phase extraction using molecularly imprinted polymer for analysis of methamidophos in water and soil samples

An analytical methodology for the analysis of methamidophos in water and soil samples incorporating a molecularly imprinted solid-phase extraction process using methamidophos-imprinted polymer was developed. Binding study demonstrated that the polymer exhibited excellent affinity and high selectivity to the methamidophos. Evidence was also found by FT-IR analysis that hydrogen bonding between the CO(2)H in the polymer cavities and the NH(2) and P=O of the template was the origin of methamidophos recognition. The use of molecularly imprinted solid-phase extraction improved the accuracy and precision of the GC method and lowered the limit of detection. The recovery of methamidophos extracted from a 10.0 g soil sample at the 100 ng/g spike level was 95.4%. The limit of detection was 3.8 ng/g. The recovery of methamidophos extracted from 100 mL tap and river water at 1 ng/mL spike level was 96.1% and 95.8%, and the limits of detection were 10 and 13 ng/L respectively. These molecularly imprinted solid-phase extraction procedures enabled selective extraction of polar methamidophos successfully from water and soil samples, demonstrating the potential of molecularly imprinted solid-phase extraction for rapid, selective, and cost-effective sample pretreatment.     

Seasonal changes of methamidophos susceptibility and biochemical properties in Plutella xylostella (Lepidoptera: Yponomeutidae) and its parasitoid Cotesia plutellae (Hymenoptera: Braconidae)

Methamidophos resistance and acetylcholinesterase (AChE) insensitivity to methamidophos, dichlorvos, and carbofuran were determined in the field populations of Plutella xylostella (L.) (Lepidoptera: Yponomeutidae) and its parasitoid Cotesia plutellae Kurdjumov (Hymenoptera: Braconidae) collected from the corresponding hosts between October 1998 and December 2003 in Fuzhou and Minhou, Fijian, China. Resistance levels to methamidophos and AChE insensitivity to the three insecticides in the two species of insects were high during autumn and spring and low during summer. Resistance to methamidophos was 15.3- and 12.6-fold higher in resistant F0 parents of P. xylostella and C. plutellae than in their susceptible F11 progeny, respectively. The bimolecular rate constant (k(i)) values of AChE to methamidophos, dichlorvos, and carbofuran were 4.6-, 6.3-, and 7.7-fold higher in F11 progeny of P. xylostella, and 3.7-, 4.5-, and 3.7-fold higher in F11 progeny of C. plutellae than those in their F0 parents, respectively. Compared with susceptible F11 progeny, the resistance ratios for methamidophos were 4.2-29.8 and 3.8-13.1 in 21 field populations of P. xylostella and C. plutellae, respectively. The k(i) values of AChE to methamidophos, dichlorvos, and carbofuran were 2.0-21.6-, 3.6-9.5-, and 2.6-9.2-fold higher in F11 progeny of P. xylostella, and 1.8-7.6-, 1.9-4.6-, and 2.2-7.6-fold higher in F11 progeny of C. plutellae than those in 21 field populations, respectively. Significant correlative variations of methamidophos resistance as well as significant correlative variations of k(i) values of AChE to insecticides between the two species of insects also were found in space and time. The k(i) values of AChE to insecticides in C. plutellae were far higher than those in P. xylostella. There were no obvious differences in the Km and Vmax of AChE between F0 parents and F11 progeny of P. xylostella and C. plutellae, respectively. But carboxylesterase activity was 1.6-fold higher in F0 parents of C. plutellae than in F11 progeny, and glutathione S-transferase activity was 1.5-fold higher in F0 parents of P. xylostella than in F11 progeny. The results suggested that the AChE insensitivity to insecticides might play the most important role in methamidophos resistance in the two species of insects. From these results, a spatial and temporal correlative evolution of methamidophos resistance and insensitive AChE was found to exist between P. xylostella and C. plutellae.     

甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态的影响

应用3天种群增长实验方法,在(25±1)℃、无光照、以3.0×106 cells/mL的斜生栅藻为轮虫的食物等条件下,研究了亚致死浓度(0.01、0.1、1.0、10.0、100.0、1000.0和10000.0 µg/L)的甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态的影响。结果显示,甲胺磷和乙酰甲胺磷显著地影响萼花臂尾轮虫的种群增长率、混交雌体数与非混交雌体数的比值和混交率。甲胺磷显著地影响萼花臂尾轮虫种群中的带卵雌体数与不带卵雌体数的比值,但乙酰甲胺磷对其无显著的影响。和对照组相比,浓度为100.0 µg/L 的甲胺磷和浓度为1.0－10,000.0 µg/L 的乙酰甲胺磷均使轮虫种群增长率显著增大,而浓度为1000.0 µg/L和10000.0 µg/L的甲胺磷却使之显著减小;1000.0 µg/L 的甲胺磷使轮虫种群中的带卵雌体数与不带卵雌体数的比值显著上升,0.1 µg/L的甲胺磷和10.0－10000.0 µg/L的乙酰甲胺磷均使轮虫种群中的混交雌体数与非混交雌体数的比值显著上升,0.1µg/L 的甲胺磷和10 µg/L的乙酰甲胺磷均使轮虫的混交率显著增大,10.0－0000.0 µg/L 的乙酰甲胺磷使轮虫休眠卵产量显著提高。上述结果表明,亚致死浓度的甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态具有显著的影响。     

褐飞虱抗甲胺磷品系的交互抗性和抗性生化机制

用甲胺磷筛选的褐飞虱Nilaparvata lugens品系（R）,对甲胺磷的抗性达到43.74倍,对马拉硫磷、二嗪磷、异丙威、仲丁威及醚菊酯都表现出一定的交互抗性,而对氰戊菊酯和吡虫啉的交互抗性不显著。为了研究褐飞虱对甲胺磷抗性和对其它药剂交互抗性产生的机制,进行了活体增效试验和离体生化实验。用2 μg/头的增效剂预处理试虫的活体增效实验结果显示,在甲胺磷筛选品系（R）中, TPP(triphenyl phosphate, 磷酸三苯酯)对甲胺磷的增效倍数达到4.54,TPP对马拉硫磷、二嗪磷、仲丁威、异丙威都表现出一定的增效作用,增效比分别为2.76、2.07、2.17和1.64;PBO(piperonyl butoxide,胡椒基丁醚)对甲胺磷、马拉硫磷和醚菊酯有一定的增效作用;DEM(diethyl meteate, 顺丁烯二酸二乙酯)的增效作用不明显。研究离体情况下增效剂对三种解毒酶活性的影响发现,TPP对R品系酯酶活力抑制作用很强（抑制率69.04%）,PBO对多功能氧化酶（MFO）具有一定的抑制作用（抑制率29.30%）,而TPP和PBO在F品系和S品系中对酯酶和MFO的抑制作用都较小;DEM在三个品系中对谷胱甘肽-S-转移酶的抑制作用都很小。由此可见,酯酶在褐飞虱对甲胺磷的抗性中起最主要作用,在马拉硫磷、二嗪磷、异丙威和仲丁威的交互抗性中起很重要作用;MFO可能在甲胺磷抗性和醚菊酯、马拉硫磷的交互抗性中起一定作用。     

Insecticide odour interference with food-searching behaviour of Microplitis croceipes (Hymenoptera: Braconidae) in a laboratory arena

Response to odours plays an important role in resource location by natural enemies, particularly by parasitoid wasps. While a considerable research effort has been dedicated to studying the effects of insecticide intoxication on natural enemy search behaviour, it is yet unknown if the odours themselves interfere with distant chemoreception. We investigated this issue using the food-searching behaviour of Microplitis croceipes (Cresson)(Hymenoptera: Braconidae) in laboratory arenas as a model system. Odours of imidacloprid (Genesis®), spinosad (Entrust®), esfenvalerate (Asana®), methamidophos (Monitor®), and vanilla were tested for their ability to interfere with wasp response to the odour of honey. The wasps did not contact the chemicals. Honey odour was generally effective in triggering food-searching behaviour in both honey-fed (and thus conditioned to associate honey odour with food) and unfed, naïve wasps. Mixing honey with imidacloprid and spinosad did not affect wasp responses. The remaining compounds (esfenvalerate, methamidophos, and vanilla) significantly reduced the proportion of positively responding fed wasps, but only methamidophos had such an effect on the unfed wasps. Negative methamidophos effects became completely reversed when wasps were forced to feed on honey in the presence of methamidophos odour. Our results suggest that odours and provision of food may potentially be used to keep beneficial natural enemies away from insecticide-treated areas.