Institute for Multidisciplinary Applied Biology Research (IMAB), Universidad Pública de Navarra, Mutilva, Spain.
Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR), Universidade do Porto, Matosinhos, Portugal.
PLoS One. 2019 Feb 7;14(2):e0210545. doi: 10.1371/journal.pone.0210545. eCollection 2019.
Quantitative laboratory bioassay methods are required to evaluate the toxicity of novel insecticidal compounds for pest control and to determine the presence of resistance traits. We used a radioactive tracer based on 32P-ATP to estimate the volume of a droplet ingested by two dipteran pests: Ceratitis capitata (Tephritidae) and Drosophila suzukii (Drosophilidae). Using blue food dye it was possible to distinguish between individuals that ingested the solution from those that did not. The average volume ingested by C. capitata adults was 1.968 μl. Females ingested a ~20% greater volume of solution than males. Adults of D. suzukii ingested an average of 0.879 μl and females ingested ~30% greater volume than males. The droplet feeding method was validated using the naturally-derived insecticide spinosad as the active ingredient (a.i.). For C. capitata, the concentration-mortality response did not differ between the sexes or among three different batches of insects. Lethal dose values were calculated based on mean ingested volumes. For C. capitata LD50 values were 1.462 and 1.502 ng a.i./insect for males and females, respectively, equivalent to 0.274 and 0.271 ng a.i./mg for males and females, respectively, when sex-specific variation in body weight was considered. Using the same process for D. suzukii, the LD50 value was estimated at 2.927 ng a.i./insect, or 1.994 ng a.i./mg based on a mean body weight of 1.67 mg for both sexes together. We conclude that this technique could be readily employed for determination of the resistance status and dose-mortality responses of insecticidal compounds in many species of pestiferous Diptera.
需要定量实验室生物测定方法来评估新型杀虫剂化合物对害虫防治的毒性,并确定抗性特征的存在。我们使用基于 32P-ATP 的放射性示踪剂来估计两种双翅目害虫——地中海实蝇(Tephritidae)和果蝇(Drosophilidae)摄入的液滴体积。使用蓝色食物染料,可以区分摄入溶液和未摄入溶液的个体。成虫摄入的平均体积为 1.968 μl。雌性摄入的溶液量比雄性多约 20%。果蝇成虫平均摄入 0.879 μl,雌性摄入的量比雄性多约 30%。使用天然衍生杀虫剂多杀菌素作为活性成分(a.i.)验证了点滴喂食法。对于地中海实蝇,性别之间或三个不同批次的昆虫之间的浓度致死反应没有差异。根据平均摄入量计算致死剂量值。对于地中海实蝇,雄性和雌性的 LD50 值分别为 1.462 和 1.502 ng a.i./昆虫,相当于雄性和雌性的体重特异性变化时分别为 0.274 和 0.271 ng a.i./mg。对于果蝇,使用相同的过程,LD50 值估计为 2.927 ng a.i./昆虫,或基于两性平均体重为 1.67 mg 时为 1.994 ng a.i./mg。我们得出结论,该技术可用于快速确定许多害虫双翅目物种中杀虫剂化合物的抗性状态和剂量致死反应。