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LC50分别为9.3284、2.1440和7.5007 μg/cm2,但杀螟硫磷的LC95为459.4949 μg/cm2,远高于敌敌畏的8.2453μg/cm2和溴氰菊酯(含增效醚)的14.5274 μg/cm2.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.     

电子束辐照对嗜虫书虱存活与繁殖力的影响

采用电子加速器产生电子束辐照处理嗜虫书虱Liposcelis entomophila(Enderlein),分别以0、50、100、200、300、400、500、1000Gy的剂量处理成虫和若虫,观察受辐照后的存活情况和成虫的繁殖情况;以0、25、50、100、200、300、400、500Gy的剂量辐照卵,观察受辐照后卵的孵化情况。结果表明:经50Gy及以上剂量辐照后嗜虫书虱的卵不能孵化,也不能存活;在300Gy及其以上剂量经过不到8周的时间后成虫和若虫都不能存活,且在300、400、500、1000Gy的剂量范围内,嗜虫书虱成虫和若虫受辐照后的剂量越大,存活率降低幅度越大。嗜虫书虱各虫态对电子束辐照的敏感性由高至低依次为卵、若虫、成虫;经300Gy辐照后的成虫和若虫分别在8周和6周内完全死亡,300Gy的剂量可作为电子束有效防治嗜虫书虱的参考剂量。所有50Gy以上剂量下都可明显降低此种害虫的繁殖力,300Gy以上剂量的处理可导致试虫零产卵。     

两种书虱微卫星富集文库的构建及比较

利用链霉亲和素与生物素之间的强亲和性原理,将链霉亲和素偶联的磁珠与微卫星探针(AC)12、(TC)12、(ATC)8、(ATG)8、(AAC)8、(ATAC)6及(GATA)6退火结合后,再亲和捕捉含接头和微卫星序列的单链书虱基因组DNA限制性酶切目的片段,经PCR扩增形成双链后进行克隆、建库。结果表明本研究成功构建了嗜卷书虱和嗜虫书虱共13个微卫星富集文库,包括6个嗜卷书虱文库,7个嗜虫书虱文库,其平均阳性克隆率为71.17%。经检测发现共得到两种书虱260个微卫星位点。这两种书虱微卫星富集文库的建立和高多态性微卫星位点的筛选将为嗜卷书虱和嗜虫书虱的种群遗传与进化、基因连锁图谱构建、分子系统发育研究等提供大量分子遗传标记,对其在实仓中的持续控制提供遗传学信息。     

嗜虫书虱性信息素的提取方法和生物活性测定方法

首次利用嗜虫书虱Liposcelis entomophila雄虫分别爬向雌、雄虫的时间确定了嗜虫书虱是雌虫分泌的性信息素,而不是雄虫分泌的聚集信息素。首次在国内外对微小个体的啮齿目昆虫嗜虫书虱L.entomophila性信息素的提取方法和生物活性测定方法进行了研究。在自制圆盘诱捕器的生物活性测定试验中,雄虫爬向雌虫平均只需要(20.00±2.44)min(N=10),而雄虫爬向雄虫的时间平均需要(70.50±6.56)min,两者差异极显著(p<0.01),说明了是雌虫分泌的性信息素。采用有机溶剂(正己烷和二氯甲烷)浸泡法和顶空气体收集方法并用两种溶剂(正己烷和二氯甲烷)进行淋洗收集嗜虫书虱未交尾雌成虫性信息素,分别用自制的圆盘诱捕器和模拟仓诱捕器和“Y”型嗅觉仪三种方法对不同方法提取的性信息素粗提物进行了生物活性测定,结果表明:顶空气体收集法得到的正己烷淋洗液在自制的圆盘诱捕器诱捕法中对雄虫的引诱活性最强,其诱集系数和诱虫数分别是46.3%±4.2%和14.0±1.0,与未交配的雌虫的相当,两者差异不显著(p>0.05),与顶空气体收集法得到的二氯甲烷淋洗液及正己烷浸泡液和二氯甲烷浸泡液的差异均显著(p<0.05)。说明顶空气体收集法用正己烷淋洗方法可以用来收集嗜虫书虱雌成虫性信息素。不同生物活性测定方法研究表明“Y”型嗅觉仪是在理想的状态下进行的选择性试验,其对正己烷淋洗液的诱集系数和诱虫数均较高,分别为55.3%±6.3%和15.0±1.8,与圆盘诱捕法相比差异不显著(p>0.05),和模拟仓诱捕法相比差异显著(p<0.05)。因此在实际应用中主要考虑利用圆盘诱捕法对嗜虫书虱进行诱捕。     

硅藻土及其混配剂对书虱的防治效果

采用拌粮法 ,用硅藻土单剂、配方 1 (硅藻土 +95 %马拉硫磷EC)、配方 2 (硅藻土 +2 . 5 %溴氰菊酯EC)、配方 3 (硅藻土 +80 %敌敌畏EC)、配方 4(硅藻土 +0 . 4%天惠虫清EC)、配方 5 (硅藻土 +5 %双氧威WP)、配方 6(硅藻土 +5 %抑太保EC)对嗜卷书虱Liposcelisbostrychophila进行防治研究 ,结果表明 :配方 4为优选配方 ,处理 2 4h后 ,3种浓度的致死率均达到 1 0 0 % ;配方 1、配方 2、配方 3、配方 5、配方 6对嗜卷书虱的致死率处理 48h后达到 1 0 0 % ;单用硅藻土处理 96h后达到 1 0 0 % ,各配方处理间差异极显著。     

不同食物对嗜卷书虱发育和繁殖的影响

通过单头饲养方法综合比较了嗜卷书虱取食９种基本食物或食物组合的发育和繁殖情况,结果表明全麦粉最有利于该虫的发育和繁殖;在全麦粉中加入少量酵母粉和脱脂奶粉后,该虫发育明显加快、存活率及繁殖力均显著提高。此外还就该虫嗜食不同食物的原因进行了探讨。     

4种昆虫病原线虫对草地贪夜蛾的致死作用

【目的】探讨昆虫病原线虫对草地贪夜蛾幼虫的侵染效果。【方法】在室内条件下采用生测试验法,测定了小卷蛾斯氏线虫AⅡ、长尾斯氏线虫X-7、夜蛾斯氏线虫SN和嗜菌异小杆线虫H06等4种昆虫病原线虫对草地贪夜蛾2龄和5龄幼虫的致死作用。【结果】小卷蛾斯氏线虫AⅡ与草地贪夜蛾数量比为50∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为92%和100%。长尾斯氏线虫X-7与草地贪夜蛾数量比为50∶1时,36 h后草地贪夜蛾2龄幼虫的死亡率为80%;在数量比30∶1时,36 h后草地贪夜蛾5龄幼虫死亡率为100%。夜蛾斯氏线虫SN与草地贪夜蛾数量比为400∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为88%和85%。嗜菌异小杆线虫H06与草地贪夜蛾在400∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为32%和67.5%。【结论】小卷蛾斯氏线虫AⅡ具有较好的草地贪夜蛾生物防治潜质,其次是长尾斯氏线虫X-7。     

氯虫苯甲酰胺对非靶标害虫褐飞虱实验种群的亚致死效应

在稻田中,氯虫苯甲酰胺是以鳞翅目幼虫为主要防治对象的新型杀虫剂,而褐飞虱 是该药剂的重要非靶标害虫.本文采用稻茎浸渍法测定氯虫苯甲酰胺对其非靶标害虫褐飞虱3龄若虫和成虫的毒力.结果表明:氯虫苯甲酰胺对褐飞虱3龄若虫和成虫的LC50分别为26.85和35.53 mg·L-1;以氯虫苯甲酰胺亚致死浓度LC10和LC25分别处理褐飞虱3龄若虫后,对当代褐飞虱雌虫寿命无显著影响,但LC25剂量处理后,当代褐飞虱雌虫产卵量显著降低45.6粒.亚致死剂量处理褐飞虱3龄若虫后,显著影响F1代褐飞虱的产卵量和雌虫寿命,雌虫产卵量分别减少43.5和72.9粒,雌虫寿命分别缩短1.35和2.87 d;两个剂量处理后F1代的各虫态发育历期均有所延长;施药后各项种群参数也发生了变化,种群内禀增长率rm分别降低12.8％和23.5％,净增殖率R0分别降低37.4％和68.7％,而世代平均历期T和种群加倍时间t均延长.表明氯虫苯甲酰胺亚致死剂量对褐飞虱种群增长具有一定的抑制作用.     

四种杀虫剂对两种书虱羧酸酯酶和乙酰胆碱酯酶的抑制作用

选用有机磷类杀虫剂(敌敌畏、毒死蜱、对氧磷)和氨基甲酸酯类杀虫剂(丁硫克百威),通过生物测定(药膜法)和生化测定(比色法)比较了嗜卷书虱和嗜虫书虱对所选药剂的敏感差异性。根据LC50可知嗜虫书虱对所选药剂比嗜卷书虱敏感。离体酶活性分析结果显示嗜卷书虱和嗜虫书虱的羧酸酯酶只对敌敌畏敏感,且嗜卷书虱比嗜虫书虱更敏感;4种药剂对乙酰胆碱酯酶均有强烈的抑制作用,同样是嗜卷书虱比嗜虫书虱敏感。乙酰胆碱酯酶的动力学研究结果和离体酶活性测定相一致。聚丙烯酰胺凝胶电泳分析显示,4种杀虫剂离体条件下对2种书虱的酯酶同工酶的抑制能力有明显差异,其中敌敌畏的抑制力最强;但对不同同功酶的抑制趋势(对大分子的抑制似乎较强)是一致的。酶的敏感性分析结果与生测结果比较表明,2种书虱的耐药力差异与其乙酰胆碱酯酶和酯酶对药剂的敏感性无关。如要弄清耐药力机制,需做进一步研究。     

山东省不同地区灰飞虱对溴氰虫酰胺的敏感性及亚致死剂量溴氰虫酰胺对灰飞虱解毒酶活性的影响

为了探究山东不同地区灰飞虱种群对溴氰虫酰胺的敏感性水平差异,在室内采用稻苗浸渍法和点滴法分别测定了溴氰虫酰胺对泰安、莱芜、鱼台(分别采自小麦和水稻)、济南和济阳6个灰飞虱种群3龄若虫和成虫的毒力,同时测定了亚致死剂量(LD 10和LD 30)溴氰虫酰胺对灰飞虱成虫体内的酯酶(ESTs)、谷胱甘肽-S-转移酶(GSTs)和多功能氧化酶(MFO)活性的影响。稻苗浸渍法结果表明,溴氰虫酰胺对6个灰飞虱种群3龄若虫的LC 50值介于8.5193~11.0524 mg/L,对成虫的LC 50值介于10.4245~12.4904 mg/L。点滴法测定结果表明,溴氰虫酰胺对6个灰飞虱种群3龄若虫的LD 50值在0.9239×10-3~1.1318×10-3μg/头之间,对成虫的LD 50值在1.0933×10-3~1.2619×10-3μg/头之间。在溴氰虫酰胺亚致死剂量(LD 10和LD 30)处理下,灰飞虱体内3种解毒酶活性均被诱导上升,其中GSTs酶活力上升最显著。结果表明,山东不同地区田间灰飞虱种群对溴氰虫酰胺的敏感性差异不大,同一地区灰飞虱种群不同虫态对溴氰虫酰胺的敏感性也无明显差异。GSTs可能会在灰飞虱对溴氰虫酰胺后续抗性形成中起作用。     

储藏物害虫嗜卷书虱对DDVP熏蒸的行为反应与致死剂量

1 引言 嗜卷书虱(Liposcelis bostrychophila)属啮虫目虱啮科。其体形微小,食性复杂,栖息场地多在粮食仓库及食品加工厂、图书馆、标本馆等地。大量发生时可造成严重损失,传播微生物,并以其尸体、排泄物等污染储藏的粮食和其它储藏物品。王进军等调查发现,在我国“双低”和“三低”贮粮的粮仓中,书虱已成为优势种群,对贮粮安全构成了威胁。使用病原真菌和细菌来防治嗜卷书虱,很难达到理想效果,即使采用菊酯类杀虫剂也难以奏效,贮粮熏蒸中普遍使用的溴甲烷(CH_3Br)和磷化氢(PH_3)对嗜卷书虱的防治效果也不很理想。利用昆虫生长调节剂防治嗜卷书虱的研究已有一些报道,但在具体应用上还存在着一些问题。气调措施可应用于控制书虱种群的增长,但书虱的抗气性发展很快,     

嗜虫书虱种群增长模型的研究

【目的】构建嗜虫书虱Liposcelis entomophila (Enderlein)在不同环境条件下的种群增长模型,为粮库书虱种群预测预报提供基础数据和理论模型。【方法】通过构建嗜虫书虱实验种群生命表,分析温度、湿度、食物破碎率和种群初始密度对嗜虫书虱种群数量动态的影响,验证种群的指数增长模型,并拟合温湿度与内禀增长率的函数关系。【结果】在温度26-34℃,湿度65%-85%,食物破碎率4%-12%的条件下,温度和湿度对发育历期有显著影响,三者对种群数量趋势指数、净增殖率、内禀增长率的影响均不显著。三者对种群增长影响的主次顺序为:温度>湿度和食物破碎率。嗜虫书虱种群增长的最适条件为温度34℃、相对湿度85%、食物破碎率12%。当种群初始密度为0.26-1.04头/cm2时,嗜虫书虱种群数量呈指数增长。【结论】在22-34℃,相对湿度65%-85%的环境中,嗜虫书虱种群数量动态与温度和湿度的关系可以拟合为Nt=N0EXP[(0.002 96T+0.021 67RH-0.037 2)t]。     

三色书虱体内共生微生物Wolbachia的wsp基因的分子检测

采用常规PCR和巢式PCR方法对三色书虱Liposcelis tricolor体内的共生微生物Wolbachia的wsp基因进行分子检测;通过Wolbachia的通用引物以及A、B亚群引物分别比较了常规PCR和巢式PCR对wsp基因扩增的灵敏性。从三色书虱体内扩增出了610bp的Wolbachia的wsp基因片段,500bp的WolbachiaA亚群的wsp基因片段和450bp的Wolbachia B亚群的wsp基因片段。扩增结果说明三色书虱被A和B两个亚群的Wolbachia混合感染;巢式PCR比常规PCR更为灵敏。     

Molecular characterization of two nicotinic acetylcholine receptor subunits from Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae)

Two nicotinic acetylcholine receptor (nAChR) subunit genes, Lbα1 and Lbα8, were isolated and characterized from psocid, Liposcelis bostrychophila Badonnel, using the rapid amplification of cDNA ends (RACE) technique. They are the first two nAChR family members isolated from the insect order of Psocoptera. The full‐length cDNAs of Lbα1 (GenBank accession number: EU871527) and Lbα8 (EU871526) consist of 2,025 and 1,763 nucleotides, respectively, and an open reading frame of 1,644 and 1,608 bp encoding 547 and 535 amino acid proteins, respectively. Both genes have typical features of nAChR family members, though they share only 56% identity in amino acid sequence. The dendrogram generated by the MEGA 3.1 program shows that the protein deduced by Lbα1 had the closest phylogenetic relationship to Agamα1 from Anopheles gambiae and Amelα1 from Apis mellifera, and Lbα8 shares the highest identity with Agamα8 from An. gambiae and Amelα8 from A. mellifera. Quantitative real‐time PCR analysis showed that Lbα1 was expressed 2.03–6.54‐fold higher than Lbα8 at the different developmental stages of L. bostrychophila. The highest expression levels of Lbα1 and Lbα8 were both detected at adult stage and the lowest were at the third and fourth nymphal stages, respectively. There was a stable and relatively low expression level for Lbα1, whereas there was a descending expression pattern for Lbα8 in the 1st through the 4th nymphal stadia. © 2009 Wiley Periodicals Inc.     

Purification and partial characterization of glutathione S-transferase from insecticide-resistant field populations of Liposcelis paeta Pearman (Psocoptera: Liposcelididae)

Enzymes that possess glutathione S-transferase (GST) activity were purified to homogeneity by glutathione-agarose affinity chromatography from three field populations of Liposcelis paeta (Pearman). These populations were collected from Nanyang city of Henan Province (NY), Wuzhou (WZ) and Hezhou (HZ) cities of Guangxi Province, China, and had different susceptibilities to dichlorvos [LC₅₀s of the NY (281.48 mg/m²), the WZ (285.07 mg/m²), and the HZ (243.52 mg/m²), respectively]. The specific activities of purified enzymes from these three populations increased 32.24-, 99.81-, and 42.52-fold, respectively. Kinetic analyses showed that the catalytic activity of purified GST from NY population towards GSH was much higher than the others, while WZ population reached the highest in V. SDS–polyacrylamide electrophoresis revealed that the purified GST had two subunits with a molecular mass of 23.31 and 20.43 kDa for NY, 53.14 and 20.13 kDa for WZ, and 50.79 and 19.42 kDa for HZ, respectively. The in vitro inhibition studies of GSTs indicated that three kinds of insecticides (chlorpyrifos, carbosulfan, and cypermethrin) and five metallic ions (Zn²⁺, Ba²⁺, Ca²⁺, Hg²⁺, Mn²⁺, and Mg²⁺) all possessed inhibitory effects on purified GST, and ethacrynic acid (EA, a specific inhibitor of GST) expressed inhibitory effects. In the bioassay, three populations of L. paeta had different susceptibilities to different insecticides, even after they were reared on diets consisting of 25% EA. The GST activities of L. paeta from different areas also showed different temperature and pH stabilities. The differences in GST among the three populations may be attributed partially to the differences in control practices for psocids between Henan and Guangxi Provinces. © 2009 Wiley Periodicals, Inc.     

Toxicological and biochemical characterizations of GSTs in Liposcelis bostrychophila Badonnel (Psocop., Liposcelididae)

Abstract: The toxicological and biochemical characteristics of glutathione S‐transferases (GSTs) in the resistant and susceptible strains of Liposcelis bostrychophila were investigated. The two resistant strains were the dichlorvos‐resistant strain (DDVP‐R) and PH₃‐resistant strain (PH₃‐R), and the resistance factors were 22.36 and 4.51, respectively. Compared with their susceptible counterparts, the activities per insect and specific activities of GSTs in DDVP‐R and PH₃‐R were significantly higher. The apparent Michaelis–Menten constant values (K_m) for 1‐chloro‐2,4‐dinitrobenzene (CDNB) were obviously lower in DDVP‐R and PH₃‐R (i.e. lower K_m values, 1.5625 mm for DDVP‐R and 0.6230 mm for PH₃‐R) when compared with their susceptible counterpart (K_m = 3.5520), indicating a higher affinity to the substrate CDNB in resistant strains. In contrast, the catalytic activity of GSTs towards CDNB in the susceptible strain was significantly higher than those in resistant strains. It was noticeable that when reduced glutathione (GSH) was used as substrate, GSTs from resistant strains both indicated a significantly declined affinity. For the catalytic activity of GSTs towards GSH, only the V_max value in DDVP‐R increased significantly compared with that from the susceptible strain, suggesting an overexpression of GST in this resistant strain. The inhibition kinetics of insecticides to GSTs in vitro revealed that dichlorvos and paraoxon possessed excellent inhibition effects on GSTs. The susceptible strain showed higher sensitivity (I₅₀ = 0.9004 mm ) to dichlorvos than DDVP‐R and PH₃‐R (higher I₅₀s, 8.0955 mm for DDVP‐R and 9.3346 mm for PH₃‐R). As for paraoxon, there was a similar situation. The resistant strains both suggested a higher I₅₀ (1.8735 mm for DDVP‐R, and 0.4291 mm for PH₃‐R) compared with the susceptible strain (0.2943 mm ). These suggested that an elevated detoxification ability of GSTs developed in the resistant strains.     

Comparative studies of acetylcholinesterase purified from three field populations of Liposcelis entomophila (enderlein) (psocoptera: liposcelididae)

Acetylcholinesterace (AChE) is known to be the major target for organophophate and carbamate insecticides and biomolecular changes to AChE have been demonstrated to be an important mechanism for insecticide resistance in many insect species. In this study, AChE from three field populations of Liposcelis entomophila (Enderlein) (Psocoptera: Liposcelididae) was purified by affinity chromatography and subsequently characterized by its Michaelis‐Menten kinetics to determine if detectable changes to AChE have occurred. Bioassays revealed that the potential resistance threat of psocids in Sichuan Province (GH) was greater than either Hubei Province (WH) or Chongqing Municipality (BB). Compared to the other two populations, the WH population possessed the highest specific activity of purified AChE. Kinetic analyses indicated that the purified AChE from GH population expressed a significantly lower affinity to the substrate and a higher catalytic activity toward acetylthiocholine iodide (ATChI) (i.e., higher K_m and V_max values) than BB and WH populations. In vitro studies of AChE suggest that five inhibitors (aldicarb, eserine, BW284C51, omethoate, and propoxur) all possess strong inhibitory effects with eserine having the strongest inhibitory effect against purified AChE. According to bimolecular rate constants (k_i), the purified AChE from GH population was least sensitive to all inhibitors except for omethoate. The differences in AChE among the three populations may be partially attributed to the differences in pesticide application and control practices for psocids among the three locations. © 2010 Wiley Periodicals, Inc.     

嗜虫书虱触角的超微结构及其在交配行为中的作用

研究嗜虫书虱触角的超微结构及其在交配行为中的作用,对于其生物学和行为学具有重要的意义。本研究利用Quanta 250型扫描电镜对嗜虫书虱触角超微结构进行研究,在Lei CAEZ4HD型解剖镜下用WPI500374型显微剪刀分别剪去雌、雄虫触角及触角不同部位后,利用Smartzoom 5型超景深显微镜观察其交配行为。研究结果表明,嗜虫书虱雌、雄虫触角形态类似,均属于线状触角,由柄节、梗节和鞭节组成,鞭节分13个亚节,成虫触角感受器主要有微毛形感受器、刺形感受器(SC1、SC2)、锥形感受器(SB1、SB2)和B9hm氏鬃毛4类感受器6种形态,绝大部分触角感受器分布在雌雄触角的背面、腹面和外侧面,两性间触角感器的类型和分布无明显差异,但数量差异明显(雌、雄虫感受器的数量分别约为8280和7240)。嗜虫书虱触角在交配行为中起到重要的作用,其中雌虫触角的梗节在交配过程中起主要作用,其次是柄节,再次是鞭节;雄虫触角的鞭节部位起着主要的作用,其次是柄节和梗节。结合其交配行为并参考其他昆虫触角感受器的研究结果,推测雌虫触角柄节和梗节上分布的B9hm氏鬃毛和雄虫触角鞭节上分布的微毛形感受器在两性识别中起着主要作用,雌、雄虫触角上分布的两种刺形感受器(SC1、SC2)和两种锥形感受器(SB1、SB2)在交配过程中有机械感受的作用。本研究结果为嗜虫书虱的行为生物学、化学生态学和电生理学的研究提供参考。