Comparison of kinetic properties of a hydrophilic form of acetylcholinesterase purified from strains susceptible and resistant to carbamate and organophosphorus insecticides of green rice leafhopper (Nephotettix cincticeps Uhler)

A hydrophilic form of acetylcholinesterase (AChE) was purified from N-methyl carbamate susceptible (SA) and highly N-methyl carbamate-resistant (N3D) strains of the green rice leafhopper (GRLH), Nephotettix cincticeps Uhler. Both of purified AChE from SA and N3D strains displayed the highest activities toward acetylthiocholine (ATCh) at pH 8.5. In the SA strain, the optimum concentrations for ATCh, propionylthiocholine (PTCh), and butyrylthiocholine (BTCh) were about 1 × 10⁻³, 2.5 × 10⁻³, and 1 × 10⁻³ M, respectively. However, in the N3D strain, substrate inhibition was not identified for ATCh, PTCh, and BTCh to 1 × 10⁻² M. The K_m value in the SA strain was 51.1, 39.1, and 41.6 μM and that in the N3D strain was 91.8, 88.1, and 85.2 μM for ATCh, PTCh, and BTCh, respectively. The K_m value in the N3D strain indicated about 1.80-, 2.25-, and 2.05-fold lower affinity than that of the SA strain for ATCh, PTCh, and BTCh, respectively. The V_max value in the SA strain was 70.2, 30.5, and 4.6 U/mg protein and that in the N3D strain was 123.0, 27.0, and 14.5 U/mg protein for ATCh, PTCh, and BTCh, respectively. The V_max value in the N3D strain was 1.75- and 3.15-fold higher for ATCh and BTCh than that in the N3D strain. However, it was 1.13-fold lower for PTCh. The increased activity of AChE in the N3D strain is due to the qualitatively modified enzyme with a higher catalytic efficiency. The bimolecular rate constant (k_i) for propoxur was 27.1 × 10⁴ and 0.51 × 10⁴ M⁻¹ min⁻¹ in the SA and N3D strain and that for monocrotophos was 0.031 × 10⁴ and 2.0 × 10⁴ M⁻¹ min⁻¹ in the SA and N3D strain. AChE from the N3D strain was 53-fold less sensitive than SA strain to inhibition by propoxur. In contrast, AChE from the N3D strain was 65-fold more sensitive to inhibition by monocrotophos than AChE from the SA strain. This indicated negatively correlated cross-insensitivity of AChE to propoxur and monocrotophos.     

Non-target toxicology of a new mosquito larvicide, trypsin modulating oostatic factor

Trypsin modulating oostatic factor (TMOF), a peptide hormone originally isolated from the ovaries of adult Aedes aegypti, is currently under commercial development as a new pesticide chemistry with a novel mode of action for the control of larval mosquitoes. The objective of the current research is to evaluate potential risks of the use of TMOF as an insecticide on non-target organisms. TMOF (YDPAP₆) was degraded in vitro (as determined by HPLC and LC/MS) to DPAP₆, PAP₆, and then AP₆ by leucine aminopeptidase, a pancreatic enzyme found in the digestive system of vertebrates. The rate of degradation of TMOF and PAP₆ was significantly greater than that of DPAP₆, while no metabolism of AP₆ was found. TMOF technical insecticide was produced on a commercial scale by recombinant yeast (heat-killed before application). The technical TMOF when administered in a single dose by gavage to male and female mice at 2000 mg dry weight/kg body weight produced no negative effects as compared to controls up to 12 days after treatment. When male and female mallard ducks were treated by gavage with 1250 mg dry weight of technical TMOF/kg body weight each day for 5 days, again no toxic effects were noted through 35 days after the last treatment. TMOF technical insecticide was also applied to the shaved skin of male and female rabbits at the rate of 2000 mg/kg for 1-2 days, with no effect. The end point observations in these in vivo experiments were mortality; changes in growth rate, behavior, body structure, and color; and possible lesions observed during necropsy. Finally, Daphnia incubated with technical TMOF in rearing water at the level of 1.0 × 10⁶ yeast cells/ml (10 mg/ml) also demonstrated no negative effects on mortality, growth, molting, time to first brood, and production of viable neonates. It appears from these studies that TMOF can be degraded by vertebrate digestive proteases and technical TMOF is not toxic to the non-target organisms examined.     

Development of resistance to insecticides in the invasive mealybug Phenacoccus solenopsis (Hemiptera: Pseudococcidae) in Pakistan

The mealybug Phenacoccus solenopsis has been a serious pest of cotton, vegetables, ornamentals and other plants since its invasion into Pakistan in 2005. Its susceptibility to commonly-used insecticides was monitored during 2005–2013 by a nymphal dip bioassay. Initially, P. solenopsis was found susceptible to a range of insecticide classes. Lethal concentration values were particularly low for organophosphates and pyrethroids. After a year of its exposure to insecticides, P. solenopsis developed moderate to high levels of resistance to pyrethroids. After two years, resistance to organophosphates methidathion and chlorpyrifos also rose to moderate to high levels. After five years of use, resistance to pyrethroids, organophosphates, neonicotinoids, endosulfan, carbosulfan, and thiocyclam was generally high to very high. Acetamiprid resistance was slow to develop, as it reached moderate level of resistance after seven years of its extensive applications. Insecticide resistance in P. solenopsis could have been managed in Pakistan if the effective and diverse insecticides were used in rotation, along with other integrated pest management tactics, at the initial stages of resistance development.     

海南不同地区香蕉假茎象甲对几种杀虫剂的敏感性测定

采用虫体浸渍法，测定了海南澄迈和儋州两地的香蕉假茎象甲成虫对480克/升毒死蜱EC、77.5%敌敌畏EC、40%辛硫磷EC、40%丙溴磷EC、25%丙溴?辛硫磷EC、20%三唑磷EC等6种杀虫剂对的敏感性。结果表明，6种药剂对澄迈和儋州两地区的香蕉假茎象甲的毒力大小顺序相同，毒死蜱＞丙溴?辛硫磷＞丙溴磷＞三唑磷＞敌敌畏＞辛硫磷；儋州地区的香蕉假茎象甲对6种药剂的敏感性均高于澄迈地区。     

一硫代磷酸酯类杀虫剂的微生物降解

在纯培养条件下测定了分离株 YF0 5对对硫磷等一硫代磷酸酯类杀虫剂的降解作用。在接种量 (OD4 15nm)为 0.2、pH7.0、30℃条件下 ,测得 10、25、50、100、200、500 mg/L对硫磷的降解符合一级动力学特征 ,其降解速率常数分别为 0.077、0.123、0.128、0.119、0.084、0 .013,高浓度对硫磷对 YF05菌有抑制作用 ;分离株 YF05在不同温度及 pH下对对硫磷的降解作用为 40℃ >30℃ >20℃ ,pH8.0 >pH7.0 >pH6 .0。同样条件下 ,YF05对杀螟硫磷和甲基对硫磷也有较好的降解作用 ,对甲基对硫磷的降解速率常数与对硫磷相仿 ,对杀螟硫磷的降解速率常数略高于对硫磷。     

北京地区家蝇乙酰胆碱酯酶不敏感基因表现型的检测

在1998～1999年对北京地区室外家蝇种群乙酰胆碱酯酶 (ACh E)的不敏感基因表现型进行了检测。以敏感品系为标准 ,海淀区 (A)、海淀区 (B)、丰台区、朝阳区家蝇种群具有的对敌敌畏不敏感基因表现型频率分别为 11%、34%、56%和24% ;对灭多威不敏感基因表现型频率为4%、4%、8%和20 %。ACh E对敌敌畏和灭多威的不敏感基因表现型 ,北京地区家蝇可分成4种 :1)对敌敌畏和灭多威均敏感型 ;2 )对敌敌畏不敏感 ,对灭多威敏感型 ;3)对敌敌畏敏感 ,对灭多威不敏感型 ;4 )对敌敌畏和灭多威均不敏感型。目前 ,在北京地区以对敌敌畏和灭多威均敏感的 ACh E表现型为主 ,其次为第二种类型 ,第三和第四种类型的频率不高。     

Cytochrome P-based resistance mechanism and pyrethroid resistance in the field Anopheles albimanus resistance management trial

The relative rates of cytochrome P⁴⁵⁰ selection in southern Mexican Anopheles albimanus populations were investigated during a 3 years indoor residual house spraying intervention with a pyrethroid (PYR) or DDT, a mosaic of organophosphate (OP)-PYR, and the annual rotation of OP-PYR-carbamate (CARB). This insecticide resistance mechanism, initially evenly spread in the mosquito population, correlated with PYR resistance during the second treated year, when cytochrome P⁴⁵⁰ contents increased in most villages of the PYR, rotation and mosaic schemes. However, by the third year, mean cytochrome P⁴⁵⁰ contents declined to susceptible levels in mosquitoes of the rotation and one mosaic group but not in the PYR-treated villages. In DDT-treated villages, a continuous decrement of cytochrome P⁴⁵⁰ levels occurred since the first treatment year, and susceptible levels were observed at the end of the intervention. Most correlations of cytochrome P⁴⁵⁰ levels and PYR resistance were lost during the third year, indicating that another mechanism evolved in resistant mosquito populations.     

Essential oil from the leaves of Cryptomeria japonica acts as a silverfish (Lepisma saccharina) repellent and insecticide

This is the first article to report the evaluation of a natural product used as an antisilverfish agent. Silverfish (Lepisma saccharina), primitive wingless insects, feed on a variety of materials, including paper, cotton, starch, and cereals. They can be a problem in libraries and other places where books, documents, and papers are stored. In this pilot study, the essential oil from leaves of Cryptomeria japonica was investigated to test its properties as a silverfish repellent and insecticide. The results from a repellency bioassay show that the essential oil significantly repelled silverfish. The repellent activity was 80% at a dosage of 0.01 mg/cm³. When silverfish were exposed to a concentration of 0.16 mg/cm³ of essential oil, they were killed within 10h. The chemical composition of essential oil, the emissions from a test chamber, and the residue left on filter papers previously soaked with the essential oil in a chamber were analyzed by gas chromatography-mass spectrometry. The components of the essential oil were found to be: elemol (18.22%), 16-kaurene (11.63%), 3-carene (9.66%), sabinene (9.37%), 4-terpineol (9.06%), β-eudesmol (5.70%), α-pinene (5.62%), and limonene (5.26%). Only some constituents of the essential oil compounds collected by solid-phase microextraction were found to be emitted in the test chamber. The main constituents were: 3-carene (21.03%), p-cymene (10.95%), limonene (9.49%), β-myrcene (9.39%), γ-terpinene (9.10%), α-terpinene (8.57%), and 4-terpineol (7.97%).     

Field evolution of insecticide resistance in the brown planthopper (Nilaparvata lugens Stål) in China

The brown planthopper (Nilaparvata lugens Stål) is one of the most destructive pests of rice crops in Asian countries including China, Vietnam, Thailand, etc. Evolution of resistance in this pest insect to isoprocarb, buprofezin, pymetrozine, imidacloprid and other neonicotinoid insecticides has been reported. In order to investigate the current status of resistance to commonly used insecticides, nine field populations of N. lugens were collected from Central China, East China and South China, and resistance to insecticides was monitored from 2009 to 2012. All the 9 field populations collected in 2012 had developed extremely high resistance to imidacloprid, with resistance ratios (RR) ranging from 209.3 to 616.6. Resistance to imidacloprid was much higher in 2012 than in 2009. The RR of thiamethoxam varied from 17.4 to 47.1, and the RR of nitenpyram varied from 1.4 to 3.7 in 2012. Of the 9 field populations, six populations showed higher resistance to nitenpyram in 2012 than in 2011. RR for buprofezin varied from 110.1 to 221.6 in 2012 whereas resistance was at a medium level (RR 20.4 ∼ 39.4-fold) in 2009. RR for pymetrozine ranged from 34.9 to 46.8 in 2012. As for isoprocarb, RR ranged from 21.7 to 38.1 in 2012. The obvious increase in resistance to widely applied insecticides indicates that insecticide resistance management strategies are urgently needed to prevent or delay further increase of insecticide resistance in N. lugens.     

几种杀虫剂对豆大蓟马的毒力测定及复配增效作用

豆大蓟马是海南冬种豇豆的重要害虫,严重影响豇豆产品质量和商品价值。本文采用离心管药膜法,测定了毒死蜱、阿维菌素、高效氯氰菊酯、甲氨基阿维菌素苯甲酸盐、吡虫啉和啶虫脒等6种杀虫剂对豆大蓟马的毒力,并测定了甲氨基阿维菌素苯甲酸盐与其它5种杀虫剂复配的增效作用及最佳配比。结果表明:供试药剂对豆大蓟马毒力(LC50)大小顺序为:甲维盐(0.000 5 g/L)阿维菌素(0.056 1 g/L)毒死蜱(0.125 3 g/L)高效氯氰菊酯(0.165 4 g/L)啶虫脒(0.228 3 g/L)吡虫啉(19.803 5 g/L);以甲氨基阿维菌素苯甲酸盐与毒死蜱复配的共毒系数为776;甲氨基阿维菌素苯甲酸盐与毒死蜱配比为1∶9时的共毒系数为1 733。本研究为开发豆大蓟马防治药剂和田间化学防治提供了基本资料。