Resistance monitoring and enzyme activity in three field populations of cowpea aphid (Aphis craccivora) from Egypt

The cowpea aphid (Aphis craccivora) is one of the most important sucking insect pests attacking certain legumes in Egypt particularly faba bean, cowpea and pea. In this study we monitored the resistance level of three field populations of A. craccivora to seven insecticides belonging to three different chemical classes (organophosphates, carbamates and neonicotinoids). The three populations were collected from three governorates in Egypt namely Dakahlia, Qalyobia and Beni Suef. Diagnostic concentrations (LC₉₀ values for susceptible strain) for each insecticide were established using a leaf dipping technique. Resistance monitoring showed that the field population from Dakahlia was highly resistant to all the tested insecticides. In a similar manner, the population from Qalyobia was also resistant to all insecticides except for fenitrothion to which only moderate resistance was observed. The field population from Beni Suef exhibited a lower level of resistance to all the seven tested insecticides.Biochemical assay showed that esterase activity in these three field populations was generally higher as the enzyme activity ratio ranged from 4.3 to 7.8 fold more than that for the susceptible strain. The activity of the other measured detoxifying enzymes (glutathione -S- transferase and mixed function oxidases) was moderate in the populations from Qalyobia and Dakahlia. Nevertheless, the enzyme activity in A. craccivora collected from Beni Suef was variable and differed slightly from the activity measured in the susceptible strain. Monitoring insecticide resistance among the three aphid populations was a proactive approach to detect any shift in insecticide efficiency. The possible occurrence of resistance in the cowpea aphid to the tested insecticides may be due to the higher activity of carboxylesterases. Further studies on the resistance mechanism to these insecticides are needed to provide insights in how to manage and delay the onset of the resistance and thus prolong the performance of insecticides against A. craccivora.     

增效剂对马尾松毛虫羧酸酯酶的抑制作用

对江西省十余个县（市）马尾松毛虫（ＤｅｎｄｒｏｌｉｍｕｓｐｕｎｃｔａｔｕｓＷａｌｋｅｒ）的测定表明，马尾松毛虫各种群对拟除虫菊酯的耐药力与羧酸酯酶活性有关；对溴氰菊酯、氰戊菊酯和氯氰菊酯耐药力强的种群，其羧酸酯酶活性亦强；应用五种增效剂对马尾松毛虫羧酸酯酶进行处理，ＳＶ＿１和ＴＰＰ表现出明显的抑制作用，且随ＳＶ＿１和ＴＰＰ剂量的增加，羧酸酯酶活性逐步降低；用相同剂量的ＳＶ＿１和ＴＰＰ处理马尾松毛虫后，羧酸酯酶均随时间表现出相似的变化规律。     

抗浓核病毒中国株家蚕品系中肠羧酸酯酶活力变化和基因表达差异的研究

 【目的】探讨家蚕中肠羧酸酯酶活力变化及羧酸酯酶基因表达差异与家蚕抗浓核病毒之间的关系，阐明其抗性的分子机理。【方法】以易感浓核病毒中国株家蚕品系华八和完全抗性品系BC8（华八的近等基因系）为材料，用BIO-TEK SYNERGY测定经口接种病毒(以下简称接种)后12 h、36 h、72 h 2个品系中肠的羧酸酯酶活力，并用实时荧光定量PCR研究接种后12 h、36 h、72 h中肠羧酸酯酶基因的表达差异。【结果】接种后12 h中肠羧酸酯酶活力BC8接种组分别是BC8对照组、华八接种组和华八对照组的3.28倍、2.26倍和3.02倍,差异达到极显著水平（P＜0.01）其它时间段的供试组之间无统计学差异；接种后12 h中肠羧酸酯酶基因的相对表达量BC8接种组分别是华八接种组、华八对照组、BC8对照组的17.714倍、21.76倍和15.09倍,差异达到极显著水平（P＜0.01），其它时间段的供试组之间也无统计学差异。【结论】接种后12 h抗性品系BC8中肠羧酸酯酶活力升高及中肠羧酸酯酶基因表达水平升高可能与家蚕品系是否有抗性基因(nsd/nsd)有关,也与浓核病毒中国株的感染刺激有关；抗性品系BC8中肠羧酸酯酶活力升高的分子基础可能是羧酸酯酶基因表达水平的改变。     

桑天牛成虫取食北京杨后血淋巴羧酸酯酶活力的变化

桑天牛成虫取食不同寄主植物以后,其寿命和产卵量明显不同。对羧酸酯酶活性测定表明,以桑树作为补充营养寄主的桑天牛成虫的血淋巴羧酸酯酶活力明显高于取食北京杨的天牛;而羧酸酯酶动力学研究表明,桑天牛取食北京杨后对其羧酸酯酶活性产生了一定的影响,因饲喂时间的不同,羧酸酯酶活性差异不同,5 d以内,酶分子的活力降低,至第7天时,除了活力降低外,酶分子的结构也发生了变化。     

A survey of mutations in the Cochliomyia hominivorax (Diptera: Calliphoridae) esterase E3 gene associated with organophosphate resistance and the molecular identification of mutant alleles

Cochliomyia hominivorax (Calliphoridae) is one of the most important myiasis-causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. In Brazil, C. hominivorax has been controlled mainly with organophosphate (OP) insecticides, although the inappropriate use of these chemicals can result in the selection of resistant flies. Changes in carboxylesterase activity have been associated with OP insecticides in some arthopodan species. In this work, we isolated and characterized part of the E3 gene in C. hominivorax (ChE7), which contained the same substitutions responsible for the acquisition of OP hydrolase activity in Lucilia cuprina (Calliphoridae). Digestion of the polymerase chain reaction products with a restriction enzyme that specifically recognized the mutation site unambiguously differentiated wild and mutated esterase alleles. The PCR-RFLP assay therefore provided a fast, reliable DNA-based method for identifying C. hominivorax individuals with a mutation in the esterase gene. Further bioassays to determine the association of this mutation with OP resistance in C. hominivorax should allow the development of more effective strategies for managing this species.     

不同水稻品种对稻水象甲羧酸酯酶活性的影响

不同水稻品种对稻水象甲Lissorbqptrus oryzqphilus Kusche1羧酸酯酶活性具有明显的影响,在试验的4个水稻品种（系）中,取食辽粳454的稻水象甲α-NA羧酸酯酶活性最高（1.20μmol/mg?min）,是取食T03的4.65倍。β-NA羧酸酯酶活性以取食辽粳294的稻水象甲最高（2.11μmol/mg?min）,是取食T03的4.48倍。     

小菜蛾对辛硫磷的抗性选育及其乙酰胆碱酯酶和羧酸酯酶活力变化

在室内模拟田间药剂的选择压力,用辛硫磷对小菜蛾（Plutella xylostella）逐代处理,以选育其抗性种群,并测定乙酰胆碱酯酶（acetylcholinesterase,AChE）和羧酸酯酶（carboxylesterase,CarE）的活性变化。选育至12代,对辛硫磷抗性增长到4.524倍。生化分析表明,抗性品系AChE和CarE活性均显著高于敏感品系。在抗性选育过程中,小菜蛾AChE酶活性增长迅速,其比活力由F0的0.035μmol/L·mg·min上升为F12的0.209μmol/L·mg·min;小菜蛾不同选育世代CarE酶活性差异显著,F12比活力增长到F0的12.944倍。     

野桑蚕羧酸酯酶基因(BmmCarE-2)的克隆及表达分析

羧酸酯酶参与昆虫对有机磷和氨基甲酸酯等杀虫剂抗性的产生。通过反转录聚合酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)方法克隆了一个野桑蚕羧酸酯酶全长基因(BmmCarE-2)。序列分析表明该基因包含1个1 623 bp的开放读码框,有57 bp的cDNA5′端非翻译区序列(5′UTR)和79 bp的cDNA3′端非翻译区序列(3′UTR),编码540个氨基酸,GenBank登录号为EU328351。序列比对分析表明BmmCarE-2与家蚕羧酸酯酶基因BmCarE-2(GenBank登录号:DQ311250)的氨基酸序列相似性最高,达98.9%。利用半定量RT-PCR进行组织表达分析表明,BmmCarE-2在野桑蚕幼虫的头部和脂肪体表达量较高,在丝腺和中肠稍低,而在血液中的表达量最低。     

接种BmNPV后抗性品种和非抗性品种家蚕组织中2种酶活性的比较研究

[目的]探讨抗性品种和非抗性品种家蚕接种家蚕核型多角体病毒(Bm NPV)后体内羧酸酯酶(Car E)和乙酰胆碱酯酶(Ach E)活性的变化规律。[方法]对抗性品种及其对照品种家蚕添食Bm NPV,测定并比较中肠和血液中羧酸酯酶和乙酰胆碱酯酶的活性变化。[结果]家蚕接种Bm NPV后,抗性品种的中肠中羧酸酯酶活性上升,而对照品种羧酸酯酶活性先升高,然后从第3天开始活性降低。抗性品种前2 d中肠的乙酰胆碱酯酶活性降低,而对照品种与之相反。家蚕接种Bm NPV后,抗性品种在第2天血液羧酸酯酶活性有所升高,此后基本保持不变,而非抗性品种在第3天羧酸酯酶活性升高后下降;抗性品种血液乙酰胆碱酯酶活性升高,对照品种前2 d乙酰胆碱酯酶升高后降低。[结论]抗性品种在感染Bm NPV病毒后其中肠中羧酸酯酶活性高于未感染的抗性品种,表明抗性品种对感染病毒做出了有效应答和防御;而在感染病毒后抗性品种的中肠与血液中乙酰胆碱酯酶活性均变化相对较小。据此推测,抗性品种在感染病毒后仍保持了相对稳定的代谢过程。     

Characterisation of the carboxylesterase enzyme cauxin in the seminal fluid of the cat

The carboxylesterase cauxin is a major urinary protein in cats that is also found in seminal fluid (SF). This study investigated cauxin in feline SF including biochemical features, concentration, distribution and gene expression in epididymal tissue, and its reaction with acylglycerol substrates.Monomeric, dimeric, and/or multimeric forms of cauxin carrying N-glycosylations were detected on Western blots of feline SF but most were monomeric. Cauxin concentrations were markedly lower in SF (0.042 ± 0.020 mg/mL) than in urine (∼0.5 mg/mL) and cauxin gene expression was 60-fold lower in the epididymis than in the kidney. Immunohistochemical examination localised cauxin within the stereocilia and cytoplasm of epithelial cells lining the caput and corpus epididymis. Cauxin-positive spermatozoa were detected in the lumen of the cauda epididymis but not in the cytoplasm of the epithelial cell lining. Using an in vitro assay, cauxin hydrolysed saturated 1-mono- but not di- and tri-acylglycerols. The results suggest that cauxin secreted from the caput and corpus epididymis acts as an esterase on lipid within feline SF.