Aerial Spraying Trials in West Africa for Blackfly Control

Abstract

Organochlorine, organophosphate (o.p.) and carbamate insecticides have been extensively used in Australia to combat Lucilia cuprina Wied. the main initiator of fly-strike of sheep occurring wherever sheep are run on the continent. The improved mules operation, mid-season crutching and insecticides are important in the management of Merino sheep. Insecticides are particularly valuable against body strike, especially in young sheep, in years with intermittent rain during the warmer months. Resistance to larvicides developed in the blowfly over a period of 10y from 1957. The time between the introduction of various insecticides and development of resistance is considered and a comparison made between the emergence of resistance problems to the newer synthetic insecticides in the housefly in Denmark, and in the blowfly in Australia. To provide a complete history of insecticides against the blowfly reference is made to arsenicals, to which there was no suspicion of resistance until low order cross resistance was diagnosed in o.p.-carbamate resistant strains. Some seven years after their introduction DDT and γ BHC were replaced by cyclodiene insecticides in 1954/1955. Reasons are advanced to explain the non-emergence of resistance to DDT and BHC in that period. Resistance to dieldrin and aldrin developed in late 1957 after which diazinon was introduced. The resistance has a typical BHC/dieldrin resistance spectrum and is due to a semi-dominant gene which has persisted in the field in the absence of pressure from cyclodiene insecticides. Non-specific resistance to o.p. insecticides developed in two steps. Low order resistance, diagnosed in 1965, was supplemented by an additional resistance mechanism in 1966, three alleles on two chromosomes are involved. The carbamate, butacarb, was effective against o.p. resistant strains of the blowfly when introduced in the 1966/67 season. In 1967, resistance to butacarb was diagnosed and rapidly became widespread. For the past three fly seasons larvae have been used to monitor resistance levels to o.p. and carbamate insecticides; resistance factors to both these classes of insecticide are significantly higher in larvae than adult females. Resistance levels to o.p. insecticides have stabilised. By contrast resistance levels to butacarb have doubled. A combined o p.-carbamate resistance generally applies in field samples. Larvae from o.p.-carbamate resistant strains form artificial strikes earlier than susceptible larvae, particularly on sheep treated with butacarb. It is concluded that registered o.p. insecticides, but not butacarb, will still give considerable protection against fly-strike, providing the maximum levels of o.p. resistance, reached in laboratory selection programmes, are not exceeded in the field. Investigation into o.p. resistance in the species suggests that the resistance mechanisms place their carriers at a disadvantage in the absence of selection pressure. Measures to minimise the amount of insecticide used against the blowfly are therefore strongly advocated—these include: the improved mules operation, mid season crutching and good animal husbandry.     

二斑叶螨抗甲氰菊酯、四螨嗪、螺虫乙酯、哒螨灵品系的微卫星DNA分析

采用微卫星标记法,对二斑叶螨（Tetranychus urticae Koch）敏感品系（S）和室内筛选的4个抗性品系（抗甲氰菊酯品系、抗四螨嗪品系、抗螺虫乙酯品系、抗哒螨灵品系）进行微卫星DNA多态性分析。结果表明,二斑叶螨4个抗性品系显示出微卫星DNA 多态性。引物Tu11在相似系数0.80处将5个二斑叶螨品系区分开。引物Tu1在相似系数0.70处将5个二斑叶螨品系区分开。微卫星DNA多态性虽不能定位抗性基因,但可结合聚类分析软件,为二斑叶螨对甲氰菊酯、四螨嗪、螺虫乙酯、哒螨灵抗性快速分子检测技术提供依据,指导二斑叶螨田间抗药性监测及抗药性治理。     

18g／L阿维菌素乳油防治棉花红蜘蛛试验

进行田间试验以明确18g/L阿维菌素乳油防治棉花红蜘蛛的效果。结果表明：18g,L阿维菌素乳油（8．1～10．8）g／hm^2能够很好的控制棉花红蜘蛛,且防治效果高于对照,对作物安全。     

朱砂叶螨抗甲氰菊酯品系选育及遗传分析

 从保留的室内相对敏感品系分出一个亚品系开始培育朱砂叶螨的甲氰菊酯抗药性品系。连续用药40代，抗性增长68.5倍。以此抗性品系（R）和相对敏感品系（S）杂交、回交，研究朱砂叶螨对甲氰菊酯的抗性遗传形式。正交（SR）和反交（RS）F1代的LC-P线介于S与R之间且偏向S方，显性度DSR和DRS分别为-0.83和-0.29，表明抗性由不完全隐性基因控制；两D值（DRS和DSR）95%置信限没有重叠，表明两个 D值（DSR和DRS）存在显著差异，也表明朱砂叶螨对甲氰菊酯的抗性遗传可能存在母体影响或核外效应；对F1杂合子与亲本的回交（SR♀×S♂和RS♀×R♂）F2代测定表明抗性由单基因控制。     

截形叶螨取食对马铃薯酶活性的影响

为明确截形叶螨(Tetranychus truncatus)取食刺激对不同耐旱型马铃薯(Solanum tuberosum)品种酶活性的影响,选择2个马铃薯品种（‘荷兰15号’,早熟,干旱敏感型品种;‘陇薯10号’,晚熟,耐旱型品种）,在人工接螨后,测定7天内马铃薯叶片5种防御酶(CAT、POD、PPO、PAL、LOX)活性的变化。CAT、POD、PPO、PAL、LOX等5种酶活性在受螨害刺激后均表现出不同程度的升高,在时序上都表现为先升高后下降的趋势,且在3天后显著高于CK（未接螨的健康植株）,5种酶均参与了植物机体对害螨的防御反应,以促进受害植株体内自由氧的消减,通过木质素、花青素、生物碱等苯丙素类化合物、氢过氧化物等含量的提高来增强机体防御能力;在螨害7天后,5种酶活性都显著下降,且显著低于CK,说明叶螨在长时间危害后可能导致植株机体紊乱,植株不能很好地持续维持高效的对螨害的防御响应。2个马铃薯品种在叶螨为害后同一时间内,从CAT、PPO和PAL活性变化角度分析,干旱敏感型马铃薯品种表现出较高的抗螨性;被叶螨取食为害后耐旱型马铃薯品种POD活性较高,但干旱敏感型马铃薯品种POD活性增长幅度更大;2个品种在LOX活性上无显著差异。总体分析可得,干旱敏感型马铃薯比耐旱型品种有更高的抗螨性。研究结果可为马铃薯叶螨的科学防治和品种选育提供理论依据。     

朱砂叶螨乙酰胆碱酯酶的表达纯化及其酶活测定条件优化

乙酰胆碱酯酶(Acetylcholinesterase,AChE)是生物体内保证神经信号正常传递的关键酶,是有机磷和氨基甲酸酯类农药的作用靶标。现研究拟采用发酵罐培养含pET-30a/ace的表达菌株E.coli BL21(DE3),大量表达纯化螨AChE,利用单因素试验和正交试验优化AChE活性的测定条件。结果表明:发酵培养的重组菌株大量表达约为68kD的朱砂叶螨AChE蛋白。底物浓度、温度、pH和酶浓度等各测定因素对朱砂叶螨乙酰胆碱酯酶活性存在显著影响。确定朱砂叶螨乙酰胆碱酯酶活性测定的最佳反应条件为:底物浓度1mmol/L,pH值7,温度35℃,酶浓度0.5mg/mL。通过体外发酵培养纯化获得了大量的朱砂叶螨AChE重组蛋白,确定了螨AChE活性测定的最佳条件,为进一步研究其酶学性质以及AChE抑制剂的体外筛选奠定基础。     

朱砂叶螨乙酰胆碱酯酶真核表达载体构建及在293T细胞中表达条件

乙酰胆碱酯酶(AChE;EC 3.1.1.7)是神经传导中调节神经递质乙酰胆碱水平的一种关键酶,是有机磷和氨基甲酸酯类农药的主要作用靶标。目前关于朱砂叶螨乙酰胆碱酯酶的真核表达还未见报道。本研究拟构建朱砂叶螨AChE的真核表达载体,进行在293T细胞中表达条件摸索。将朱砂叶螨野生型ace完整读框和去掉3’疏水区的ace基因分别插入到真核表达载体pcDNA3.1/V5-His-TOPO中,经PCR和双酶切鉴定及测序验证后,成功构建了真核表达载体pcDNA3.1/ace。采用GFP报告基因确定在293T细胞中表达条件摸索,结果表明293T细胞的最适转染时间为48h和LipofectamineTM2000转染试剂的最适加样量为20μL。本试验成功构建了朱砂叶螨AChE的真核表达载体,确定了在293T细胞中的表达条件,为后续筛选新型选择性AChE抑制剂奠定了基础。     

Olfactory response of the ladybird beetle <Emphasis Type="Italic">Stethorus gilvifrons</Emphasis> to two preys and herbivore-induced plant volatiles

The spider mites Tetranychus urticae Koch and Panonychus ulmi (Koch) (Tetranychidae) cause severe economic losses to vegetable farms and deciduous fruit orchards in Turkey. One of their predators, the ladybird beetle Stethorus gilvifrons (Muls.) (Col., Coccinellidae), aggregates on mite-infested patches of plants. The present study assessed whether there is a role for herbivore-induced plant volatiles (HIPVs) and/or odors emitted directly from these two mite species in the aggregative response of ladybird beetles. The olfactory responses of the predator females to volatiles from T. urticae- and/or P. ulmi-infested sweet pepper (four cultivars, viz. ‘Demre’, ‘Yalova Carliston’, ‘Kandil Dolma’ and ‘Yag Biberi’), kidney bean (cv. ‘Barbunya’) and apple (M9 rootstock) were investigated using a two-choice olfactometer. Our results showed that HIPVs emitted from both T. urticae- and P. ulmi-infested plants significantly attracted S. gilvifrons adults for all plants except the sweet pepper cv. Yag Biberi. In addition, it was found that volatiles from apple plants infested by T. urticae and, especially, P. ulmi are more attractive for S. gilvifrons females than those emitted by other infested plants. The results also suggest that the odors of T. urticae adults and their products might influence the attraction of S. gilvifrons females.     

9种杀螨剂对朱砂叶螨不同发育阶段的室内毒力测定

室内采用FAO推荐的玻片浸渍法和叶碟法分别测定了9种杀螨剂对朱砂叶螨雌成螨和卵的毒力。结果表明,杀螨活性最高的是阿维菌素,其次是溴虫腈,卡死克、噻螨酮、哒螨灵、三唑锡也具有较好的杀螨效果,炔螨特的杀螨效果较差,甲氰菊酯和氧化乐果的杀螨效果最差;阿维菌素对朱砂叶螨卵的毒力最高,毒力次之的是噻螨酮、哒螨灵、溴虫腈、三唑锡和卡死克,甲氰菊酯、炔螨特和氧化乐果对卵的毒力最差。另外,对朱砂叶螨雌成螨和卵毒力相差最大的是噻螨酮和哒螨灵,毒力分别相差150.27和134.28倍;其次为三唑锡,相差69.80倍;溴虫腈、卡死克、阿维菌素、氧化乐果、甲氰菊酯、炔螨特对卵和雌成螨的毒力差别相对较小。     

假蒟提取物对皮氏叶螨的生物活性测定

通过索氏抽提和乙醇浸提法提取假蒟(Rper.saormentosum Roxb.)杀虫活性成分,即获得乙醇浸提物、乙醇索氏提取物和石油醚索氏提取物,研究其对皮氏叶螨(Tetranychus pieroei McGregor)各龄虫的触杀及对成螨驱避、产卵抑制等方面的生物活性.结果表明:乙醇浸提物和石油醚索氏提取物对皮氏叶螨卵的毒杀效果均较好,用20mg/g的浓度处理后,杀卵效果均在95%以上:对若螨的生物活性以乙醇索氏提取物最好,当浓度为15mg/g时,校正死亡率达95.19%,明显强于石油醚索氏提取物;乙醇浸提物、乙醇索氏提取物和石油醚索氏提取物对成螨的LC50分别为3.2141,6.093 1,3.518 8 mg/g,LC95分别为8.5750,18.700 7,9.311 4 mg/g;用质量分数0.4%乙醇浸提物和石油醚索氏提取液处理的皮氏叶螨成螨寿命明显短于乙醇索氏提取物和对照,产卵抑制指数以石油醚索氏提取物最高,为0.1465,乙醇索氏提取物最低,为0.0825;3种提取物对皮氏叶螨均有良好的驱避作用.