嗜卷书虱和嗜虫书虱酯酶性质的比较研究

对嗜卷书虱Liposcelis bostrychophila和嗜虫书虱L. entomophila羧酸酯酶及乙酰胆碱酯酶(AChE)的生物化学性质进行了初步研究。结果表明:嗜虫书虱的羧酸酯酶活性明显高于嗜卷书虱,嗜卷书虱和嗜虫书虱羧酸酯酶对底物α-NA的Km值分别为0.665 7、0.285 7 mmol/L,Vmax值分别为1.672 2和2.463 1 mmol·(mg·30 min)-1。嗜卷书虱和嗜虫书虱乙酰胆碱酯酶的活性差异不显著,嗜虫书虱略高于嗜卷书虱。通过聚丙烯酰胺凝胶电泳,检测出8条嗜卷书虱、7条嗜虫书虱酯酶同工酶谱带,嗜卷书虱的酶谱分布比嗜虫书虱广。     

磷化氢熏蒸处理对嗜卷书虱不同虫态的致死作用

实验室条件下系统研究了磷化氢(PH3)对储物害虫嗜卷书虱Liposcelis bostrychophila卵、各龄若虫和成虫的致死作用,并选用PH3间歇熏蒸以及PH3与气调交替处理等措施对嗜卷书虱进行处理,比较了不同处理措施对嗜卷书虱种群的控制效果。结果表明,PH3熏蒸处理对嗜卷书虱各虫态有不同的致死效果。对卵而言,24、72和120 h熏蒸处理的LC50分别为0.137、0.045和0.035 mg/L;而24 h熏蒸处理对若虫的LC50在4.285~7.364 μg/L之间,对成虫的LC50为20.404 μg/L;采用25 μg/L的PH3进行24 h熏蒸处理,间隔10 d后再分别进行第2次和第3次熏蒸处理,可以完全控制嗜卷书虱的发生。采用PH3 (12 μg/L)和气调(体积比例为35% CO2,1% O2,64% N2)交替处理能够延缓嗜卷书虱种群抗性的发展,交替处理3~5次可以完全控制嗜卷书虱的发生。     

吡丙醚和烯虫酯对嗜卷书虱控制作用的研究

采用饲料混毒法系统测定了吡丙醚和烯虫酯两种昆虫生长调节剂(IGRs)对重要的储藏物害虫嗜卷书虱Liposcelis bostrychophila Badonnel若虫和成虫的致死作用,对若虫发育情况、成虫繁殖力和卵孵化率的影响。结果表明,以吡丙醚和烯虫酯10 mg/kg的混毒饲料饲喂嗜卷书虱2龄幼虫49 d后,若虫的死亡率分别为80.00%和31.11%;取食20、10和5 mg/kg吡丙醚的若虫最长存活天数分别为71.0、98.2和116.4 d,而对照仅11.3 d即全部羽化为成虫;取食20 mg/kg以下烯虫酯混毒饲料的若虫一部分仍可羽化成成虫,而且成虫仍可繁殖后代;取食吡丙醚混毒饲料的若虫,60 d后存活下来的若虫大多以超龄若虫的形式存在,而取食烯虫酯的若虫则很少出现超龄若虫。成虫取食烯虫酯(200 mg/kg)和吡丙醚(40 mg/kg)混毒饲料后7 d,死亡率分别为26.67%和23.33%,成虫产卵量分别是对照的39.77%和26.13%,产卵后15 d,卵的孵化率分别是65.71%和8.69%,而对照仅为93.18%。该项研究表明,采用40 mg/kg吡丙醚和200 mg/kg的烯虫酯对嗜卷书虱虫口数量有明显的控制作用。     

8种植物精油对嗜卷书虱的杀虫活性及香青兰精油的GC-MS成分分析

为筛选新型环保植物源仓储害虫杀虫剂,采用水蒸汽蒸馏法提取8种植物精油,滤纸药膜法、密闭空气熏蒸法测定精油对嗜卷书虱成虫的触杀、熏蒸活性,筛选出高活性植物,并用气相质谱-色谱联用仪（GC-MS）对高活性植物精油进行化学成分的分析和鉴定。结果表明：香青兰、窃衣、啤酒花精油对嗜卷书虱具有较强的触杀活性,LD₅₀分别为157.43、349.34和633.07µg/cm²;香青兰、蒌蒿和窃衣精油对嗜卷书虱具有较好的熏蒸活性,LC₅₀分别为0.20、0.70和1.66 mg/L。香青兰精油经GC-MS分析,检测并鉴定到23个化学成分,相对含量占总含量的84.22%,其主要成分为乙酸香叶酯（16.26%）、丁香酚（13.90%）、香叶醛（13.20%）、草蒿脑（7.89%）、丙烯酸-2-乙基己酯（8.55%）以及橙花醛（7.60%）。研究结果表明香青兰精油对嗜卷书虱的生物活性较好,值得进一步研究。     

不同书虱品系的磷化氢半数击倒时间与抗性相关性研究

参照FAO推荐方法测定了磷化氢对嗜卷书虱(Liposcelis bostrychophila(Badonnel))2个品系、嗜虫书虱(L.entomophila(Enderlein))3个品系和小眼书虱(L.paeta(Pearman))3个品系等共8个不同来源书虱的毒力和抗性系数Rf,测定了磷化氢浓度为2mg/mL条件下供试各书虱品系的半数击倒时间KT50,分析了其LC50值与KT50值的相关性。结果为:3种书虱的8个品系对磷化氢的抗性差异明显,高抗性的嗜虫书虱的磷化氢抗性比敏感的嗜卷书虱高1000倍以上,各品系书虱的KT50值与Rf值呈正相关,相应LC50值之对数与KT50值的对数呈直线相关。结果表明,磷化氢对书虱成虫的半数击倒时间KT50值可以用来判断书虱抗性大小,并可用于快速判断抗性以指导现场熏蒸。     

嗜卷书虱和嗜虫书虱体内磷酸酯酶的生化与毒理学特性比较

采用药膜法对嗜卷书虱和嗜虫书虱进行生物测定,从5种杀虫剂对两种书虱的LC50可以看出,嗜卷书虱对两种拟除虫菊酯类药剂较嗜虫书虱敏感,嗜虫书虱对丁硫克百威较为敏感,而对有机磷类药剂的作用两种书虱的敏感性相似.两种书虱体内酸性磷酸酯酶(ACP)和碱性磷酸酯酶(ALP)的活力比较结果表明:嗜卷书虱和嗜虫书虱体内ACP的比活力为分别14μmol/mg·30 min和2.2 μmol/mg·30 min,ALP的比活力分别为0.092μmol/mg·30 min和0.046μmol/mg·30 min.经方差分析,两种书虱ACP和ALP的比活力之间均存在显著性差异(p<0.05).动力学参数比较结果显示,两种书虱ACP的Vmax值之间的差异达到显著水平,而ALP的Km值之间存在显著性差异.离体试验结果表明,5种药剂对磷酸酯酶的作用各不相同,个别药剂在低浓度下表现诱导作用,说明磷酸酯酶在不同外源毒物的解毒代谢过程中所起的作用有所不同.     

嗜卷书虱和嗜虫书虱体内磷酸酯酶的生化与毒理学特性比较研究

采用药膜法对嗜卷书虱和嗜虫书虱进行生物测定,从5种杀虫剂对两种书虱的LC50可以看出,嗜卷书虱对两种拟除虫菊酯类药剂较嗜虫书虱敏感,嗜虫书虱对丁硫克百威较为敏感,而对有机磷类药剂的作用两种书虱的敏感性相似。两种书虱体内酸性磷酸酯酶（ACP）和碱性磷酸酯酶（ALP）的活力比较结果表明：嗜卷书虱和嗜虫书虱体内ACP的比活力为分别14 μmol/mg&#8226;30 min和2.2 μmol/mg&#8226;30 min,ALP的比活力分别为0.092 μmol/mg&#8226;30 min和0.046 μmol/mg&#8226;30 min。经方差分析,两种书虱ACP和ALP的比活力之间均存在显著性差异(p<0.05)。动力学参数比较结果显示,两种书虱ACP的V _max值之间的差异达到显著水平,而ALP的K _m值之间存在显著性差异。离体试验结果表明,5种药剂对磷酸酯酶的作用各不相同,个别药剂在低浓度下表现诱导作用,说明磷酸酯酶在不同外源毒物的解毒代谢过程中所起的作用有所不同。     

温度、湿度对嗜虫书虱生长发育和繁殖的影响

嗜虫书虱以全麦粉及酵母粉作饲料时,其各期发育速率与温度呈逻辑斯蒂曲线关系,世代发育起点和有效积温分别为15.16℃,306.75日度。在20—35℃范围内,世代存活率、种群内禀增长率以及成虫产卵量较高,高低温区均呈下降趋势。理论上29.55℃时周限增长率(λ)最大,达1.0628倍/天。高湿有利于其生长发育和繁殖,湿度低于60％时,各虫态均不能发育存活。结果表明,嗜虫书虱生长发育、繁殖最适温区为28—30℃,最适相对湿度在80％左右。     

几种杀虫剂对嗜虫书虱的触杀作用

采用滤纸药膜法比较了常用8种杀虫剂对嗜虫书虱Liposcelis entomophila的触杀作用,在药剂的推荐浓度下有机磷类杀虫剂对嗜虫书虱的急性触杀作用强于其它药剂。同时测定了杀螟硫磷、敌敌畏和溴氰菊酯(含增效醚)3种药剂对嗜虫书虱的触杀毒力,其24h内触杀作用的LC_(50)分别为9．3284、2．1440和7．5007μg/cm~2,但杀螟硫磷的LC_(95)为459．4949μg/cm~2,远高于敌敌畏的8．2453μg/cm~2和溴氰菊酯(含增效醚)的14．5274μg/cm~2。此外,48h内敌敌畏等有机磷杀虫剂对嗜虫书虱的毒杀速度较其它供试药剂快。     

斜纹夜蛾对茚虫威的抗药性汰选及交互抗性测定

为评估茚虫威抗性风险,在室内进行了斜纹夜蛾对茚虫威的抗性选育和交互抗性测定。经过10代6次室内抗性选育,获得了斜纹夜蛾对茚虫威抗性种群,与选育前相比,斜纹夜蛾对茚虫威的敏感性降低了15.63倍。抗性风险评估结果表明,斜纹夜蛾具有对茚虫威产生高水平抗性的风险。交互抗性测定发现,辛硫磷、高效氯氰菊酯和氟虫腈对茚虫威抗性种群的LC50值分别是同源对照种群的1.53、2.42和1.53倍,溴虫腈和灭多威对茚虫威抗性种群的LC50值分别是同源对照种群的0.78和0.96倍,表明茚虫威抗性种群对这几种杀虫剂未产生交互抗性。     

Influence of concentration, temperature and humidity on the toxicity of phosphine to the strongly phosphine-resistant psocid Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae)

BACKGROUND: The psocid Liposcelis bostrychophila Badonnel, is a widespread, significant pest of stored commodities, has developed strong resistance to phosphine, the major grain disinfestant. The aim was to develop effective fumigation protocols to control this resistant pest. RESULTS: Time to population extinction of all life stages (TPE) in days was evaluated at a series of phosphine concentrations and temperatures at two relative humidities. Regression analysis showed that temperature, concentration and relative humidity all contributed significantly to describing TPE (P<0.001, R(2)=0.95), with temperature being the dominant variable, accounting for 74.4% of the variation. Irrespective of phosphine concentration, TPE was longer at lower temperatures and high humidity (70% RH) and shorter at higher temperatures and low humidity (55% RH). At any concentration of phosphine, a combination of higher temperature and lower humidity provides the shortest fumigation period to control resistant L. bostrychophila. For example, 19 and 11 days of fumigation are required at 15 degrees C and 70% RH at 0.1 and 1.0 mg L(-1) of phosphine respectively, whereas only 4 and 2 days are required at 35 degrees C and 55% RH for the same respective concentrations. CONCLUSIONS: The developed fumigation protocols will provide industry with flexibility in application of phosphine.     

Toxicological and biochemical characterizations of AChE in Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae)

The toxicological and biochemical characteristics of acetylcholinesterases (AChE) in the resistant and susceptible strains (SS) of Liposcelis bostrychophila were investigated. The two resistant strains were the dichlorvos-resistant strain (DDVP-R) and the phosphine-resistant strain (PH₃-R) with resistance ratios of 22.36 and 4.51, respectively. Compared to their susceptible counterpart, the AChE activity per insect and the specific activity of AChE in DDVP-R and PH₃-R were significantly higher. There were also significant kinetic differences between DDVP-R and PH₃-R. The apparent Michaelis-Menten constant (K_m) for acetylthiocholine iodide (ATChI) was obviously lower in SS than that in PH₃-R, indicating a higher affinity to the substrate ATChI in the susceptible strains. The affinity for the substrate ATChI in DDVP-R and SS were not significantly different. The V_max value of the PH₃-R was significantly greater when compared to the V_max for the SS suggesting a possible over expression of AChE in this resistant strain. The inhibition of AChE to insecticide exposure in vitro revealed that all six insecticides were inhibitory for the extracted AChE’s. Based on the I₅₀ values, AChE of the SS were more sensitive to dichlorvos, paraoxon-ethyl, malaoxon and demeton-S-methyl than those of the two resistant strains. As for carbaryl and eserine, the PH₃-R suggested a significantly higher I₅₀s compared to the susceptible strain, while, no significant differences were found between SS and DDVP-R.     

小菜蛾对丁烯氟虫腈的抗性遗传力及风险评估

采用丁烯氟虫腈对小菜蛾敏感品系进行室内抗性筛选,饲养15代,汰选13代,平均成活率为35.12%,获得抗性品系(R),与敏感品系(S)比较,抗性指数为90.27倍。应用域性状分析法,研究小菜蛾对丁烯氟虫腈的抗性现实遗传力,并对不同杀死率下抗性发展速率进行预测。结果表明,连续筛选前7代(F₀~F₆),小菜蛾对丁烯氟虫腈的抗性现实遗传力为0.2332;不连续筛选后6代(F₇~F₁₄,8代中有2代未筛选),现实遗传力为0.0203。整个13代筛选期间,现实遗传力为0.2206。假设遗传力为室内筛选估算值的一半、死亡率为50%~90%,预计小菜蛾对丁烯氟虫腈抗性增长10倍,需要约22.8~10.3代。     

朱砂叶螨对两种杀螨剂的抗性遗传力及风险评估

在室内抗性培育的基础上,应用数量遗传学中的域性状分析法研究了朱砂叶螨对甲氰菊酯和阿维菌素两种杀螨剂的抗性现实遗传力,并对两种药剂在不同杀死率下,朱砂叶螨抗性发展的速率进行了预测。结果表明,用甲氰菊酯、阿维菌素分别连续汰选16、18代后,朱砂叶螨对两者的抗性分别为28.61和4.36倍,抗性现实遗传力分别为0.2685和0.1385。在室内选择条件下,杀死率为50％～90％时,要获得10倍抗性,甲氰菊酯需要约13～6代,阿维菌素需要约28～13代。生物源农药阿维菌素的抗性风险明显低于菊酯类药剂甲氰菊酯。试验结果为朱砂叶螨抗性治理提供了理论依据。     

四种杀虫剂对两种书虱谷胱甘肽-S-转移酶和超氧化物歧化酶的影响

谷胱甘肽-S-转移酶（GSTs）能够参与多种类型杀虫剂的解毒代谢，如有机磷类、氨基甲酸酯类、拟除虫菊酯类和有机氯类。其介导的抗性与一种或多种GSTs的过量表达有关。超氧化物歧化酶（SOD）的功能是清除超氧阴离子自由基而形成H2O2，H2O2能与O2^-形成毒性更强的HO^-。书虱是一类个体微小、行动活泼的重要储藏物害虫，书虱对化学药剂的抗性发展很快，目前已发展为储粮中的害虫优势种群，但有关书虱体内酶系的研究较少。     

Purification and biochemical characterization of glutathione S-transferases from three strains of Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae): Implication of insecticide resistance

Glutathione S-transferases (GSTs) catalyzing the conjugation of reduced glutathione (GSH) to a vast range of xenobiotics including insecticides were investigated in the psocid Liposcelis bostrychophila Badonnel. GSTs from susceptible and two resistant strains (DDVP-R for dichlorvos-resistant strain and PH₃-R for phosphine-resistant strain) of L. bostrychophila were purified by glutathione-agarose affinity chromatography and characterized by their Michaelis-Menten kinetics towards artificial substrates, i.e., 1-chloro-2,4-dinitrobenzene (CDNB), in a photometric microplate assay. The specific activities of GSTs purified from two resistant strains were significantly higher than their susceptible counterpart. For the resistant strains, GSTs both showed a significantly higher affinity to the substrate GSH while a declined affinity to CDNB than those of susceptible strain. The inhibitory potential of ethacrynic acid was very effective with highest I₅₀ value (the concentration required to inhibit 50% of GSTs activity) of 1.21 μM recorded in DDVP-R. Carbosulfan also exhibited excellent inhibitory effects on purified GSTs. The N-terminus of the purified enzyme was sequenced by Edman degradation, and the alignment of first 13 amino acids of the N-terminal sequence with other insect GSTs suggested the purified protein was similar to those of Sigma class GSTs.