Expression of insect α6-like nicotinic acetylcholine receptors in Drosophila melanogaster highlights a high level of conservation of the receptor:spinosyn interaction

Insecticide research has often relied on model species for elucidating the resistance mechanisms present in the targeted pests. The accuracy and applicability of extrapolations of these laboratory findings to field conditions varies but, for target site resistance, conserved mechanisms are generally the rule rather than the exception (Perry et al., 2011). The spinosyn class of insecticides appear to fit this paradigm and are a pest control option with many uses in both crop and animal protection. Resistance to spinosyns has been identified in both laboratory-selected and field-collected pest insects.Studies using the model insect, Drosophila melanogaster, have identified the nicotinic acetylcholine receptor subunit, Dα6 as an important target of the insecticide spinosad (Perry et al., 2007, Watson et al., 2010). Field-isolated resistant strains of several agricultural pest insects provide evidence that resistance cases are often associated with mutations in orthologues to Dα6 (Baxter et al., 2010, Puinean et al., 2013).The expression of these receptors is difficult in heterologous systems. In order to examine the biology of the Dα6 receptor subunit further, we used Drosophila as a model and developed an in vivo rescue system. This allowed us to express four different isoforms of Dα6 and show that each is able to rescue the response to spinosad. Regulatory sequences upstream of the Dα6 gene able to rescue the resistance phenotype were identified. Expression of other D. melanogaster subunits revealed that the rescue phenotype appears to be Dα6 specific. We also demonstrate that expression of pest insect orthologues of Dα6 from a variety of species are capable of rescuing the spinosad response phenotype, verifying the relevance of this receptor to resistance monitoring in the field. In the absence of a robust heterologous expression system, this study presents an in vivo model that will be useful in analysing many other aspects of these receptors and their biology.     

Acaricidal activity of spinosad and abamectin against two-spotted spider mites

Spinosad is a bioinsecticide with a high degree of selective toxicity towards insects of different orders, but its toxicity towards the two-spotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae) is under debate. In this study, we compared the acaricidal properties of spinosad with the commercial bioacaricide abamectin on the life stages of TSSM. Adulticide and ovicide bioassays were performed on a susceptible laboratory strain using direct spraying of leaf disks with five rates of spinosad (20, 25, 30, 35 and 40 mg/l), five rates of abamectin (0.125, 0.25, 0.5, 1 and 2.5 mg/l), sublethal concentrations or a combination of spinosad and abamectin. Both adulticidal and ovicidal effects of spinosad against T. urticae in the laboratory were apparent, based on morality rates of the adults, reduction of female fecundity and death of offspring. Abamectin was also found to significantly reduce female fecundity and killed offspring when applied directly on the eggs. Interestingly, sublethal concentrations of spinosad reduced female fecundity stronger than abamectin. When a mixture of spinosad and abamectin was applied at LC_50, mortality was 74%, fecundity reduction was comparable to abamectin alone and egg hatching rate was lower than by either compound alone. In conclusion, spinosad was more harmful than abamectin for TSSM life stages and the combined application is recommended.     

Lethal and sublethal effects of the naturally derived insecticide spinosad on parasitoids of Spodoptera frugiperda (Lepidoptera: Noctuidae)

Laboratory studies were performed on the lethal and sublethal effects of spinosad on three important species of parasitoids attacking Spodoptera frugiperda (J.E. Smith) in Mexico. Reproduction of the braconid Chelonus insularis (Cresson), on treated egg masses was completely eliminated at 200 parts per million (ppm) and reduced by ∼70% at 20 ppm compared to the controls. Adult C. insularis did not avoid contact with residues on maize (200 ppm), but suffered a 7-day reduction in longevity after contact with residues. Initial toxicity of spinosad applied to a natural host of S. frugiperda was concentration dependent and resulted in 23 to 100% mortality of the eulophid Euplectrus plathypenae Howard at 25 to 200 ppm, respectively. The survival of Eu. plathypenae was initially reduced, especially in males, following contact with field weathered residues on maize (200 ppm). However, survival of both sexes rapidly returned to control values on foliage sampled after rainfall. A similar effect was observed in the mortality response of female Eu. plathypenae exposed to residues on sorghum. The ichneumonid Eiphosoma vitticolle Cresson did not avoid reproduction in S. frugiperda larvae that were externally contaminated with 200 ppm spinosad, although all spinosad-treated hosts died before the parasitoid progeny could develop. We use these results to predict the impact of spinosad applications on the foraging and reproduction of these parasitoids in the field. Such predictions require validation by field studies.     

Residual contact vial method for the rapid on-site detection of insecticide resistance in Thrips palmi

A residual contact vial plus water (RCVpW) bioassay method, in which water was supplemented to minimize control mortality, was established to monitor insecticide resistance in field populations of the melon thrips, Thrips palmi. In the RCVpW, median lethal doses (LD₅₀) of six insecticides commonly used in T. palmi control, were determined at 8 h post-treatment, using a susceptible RDA strain according to the RCVpW protocol. Diagnostic doses for on-site resistance monitoring of the six insecticides, which were determined as doses two-fold higher than required to achieve LD₉₀ in the RDA strain, were in the range of 0.299 to 164.3 μg^?1 cm². Insecticide resistance levels in five field populations of T. palmi were evaluated to test the applicability of RCVpW in monitoring the pest. Although the RDA strain exhibited 100% mortality to diagnostic doses, field populations showed a reduced mortality in response to all test insecticides, indicating different degrees of resistance. In particular, all test field populations exhibited a significantly low mortality in response to spinosad, suggesting a wide distribution of spinosad resistance. Synergistic bioassay revealed that cytochrome P450-mediated metabolic factor is involved in spinosad resistance in the Korean population. Interestingly, an apparently reduced mortality to emamectin benzoate and chlofenapyr was observed in some field populations, perhaps suggesting uneven distribution of resistance to these insecticides in field populations. Our study showed that the RCVpW protocol can be employed both as an on-site resistance monitoring method for major thrip species, and in the selection of appropriate insecticides for their control.     

新型生物农药-多杀菌素

多杀菌素是由刺糖多孢菌(Saccharopolyspora spinosa)经有氧发酵后的次级代谢产物,是一种新颖的大环内酯类抗生素,兼有生物农药的安全性和化学农药的速效性。本文主要就多杀菌素的研究历史、结构、理化性质、生物合成、作用机制、杀虫活性和非靶标生物毒性及降解途径进行了综述。     

Lethal and sublethal effects of spinosad-based GF-120 bait on the tephritid parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae)

Experiments were performed to determine the likely compatibility of spinosad-based GF-120 applications and mass-releases of the braconid parasitoid Diachasmimorpha longicaudata (Ashmead) for control of tephritid fruit flies. Severe effects on D. longicaudata survival and reproduction in the laboratory contrasted with mild effects in semi-field studies. Topical application and contact with dried residues on plastic surfaces in the laboratory generally resulted in high mortality of wasps and significantly reduced survival times compared to controls. Survival times of both sexes were more severely affected when exposure to dried residues on mango leaves compared to wasps that were offered GF-120 mixed with honey. Brief (24 h) exposure to GF-120 in honey or dried residues on mango leaves did not affect total progeny production by wasps, whereas continuous exposure over a 10 d period resulted in reductions in female survival, progeny production and net fecundity (∑L_xM_x) compared to honey-fed controls. During the rainy season, wasp survival times in field cages with young mango trees that had been sprayed with GF-120 or treated with GF-120 mixed with honey were only reduced by 1 or 2 days, respectively, compared to average 10.1 d survival of honey-fed controls. During the dry season, wasp mortality was not significantly affected by exposure to GF-120 residues on mango leaves that had been subjected to natural weathering in the field, compared to controls. Our results suggest that applications of GF-120 are unlikely to adversely influence the effectiveness of mass-releases of D. longicaudata. This prediction requires validation from comparative field studies on parasitoid populations in treated and untreated areas.     

Evaluation of biorational insecticides and DNA barcoding as tools to improve insect pest management in lablab bean (Lablab purpureus) in Bangladesh

Lablab bean (Lablab purpureus) is a popular vegetable crop in Bangladesh, but farmers growing this crop experience significant losses to insect pests despite heavy reliance on conventional insecticides. We conducted field studies to improve pest management in lablab bean by testing biorational insecticides as alternatives to conventional insecticides for the control of pod borers (Maruca vitrata) and aphids (Aphis craccivora), and characterizing flower-inhabiting thrips as an emerging pest in this crop. In field experiments, spinosad was the most promising biorational we tested, suppressing pod-boring caterpillars more effectively than thiamethoxam or quinalphos. In contrast, azadirachtin (neem) did not significantly suppress any of the insect pests we measured, although target aphid populations were generally low at our research site. Using DNA barcoding at the coxI locus combined with morphological identification, we found eight thrips taxa inhabiting lablab bean flowers, dominated by Megalurothrips usitatus and M. distalis/peculiaris. A preliminary regression analysis indicated that these flower-inhabiting thrips reduced lablab bean yield. Our results suggest that spinosad may be useful within lablab bean IPM programs, and that these programs will likely need to incorporate tactics against thrips to effectively protect yield. Finally, we found that DNA barcoding was a valuable tool for pest identification in an understudied crop and region, but that to reach its full potential will require an investment in more comprehensive reference libraries.     

Toxicity and kinetics of spinosad in different developmental stages of the endoparasitoid <Emphasis Type="Italic">Hyposoter didymator</Emphasis> (Hymenoptera: Ichneumonidae) and its host <Emphasis Type="Italic">Spodoptera littoralis</Emphasis> larvae (Lepidoptera: Noctuidae)

The toxicity of spinosad was evaluated using the RaPID Assay^® Spinosad immunosorbent assay in different developmental stages of the parasitoid, Hyposoter didymator, and in its host, fourth-instar larvae of the cotton leafworm Spodoptera littoralis. Spinosad was applied directly to pupae and adults of H. didymator (ingestion or topical application) or to the immature stages of the parasitoid via the host larvae. Low amounts of spinosad were recovered from S. littoralis host larvae after topical treatment, and the compound was mainly retained in the hemolymph. Amounts of spinosad detected in third-instar larvae of H. didymator, pulled out from the hemolymph of parasitized S. littoralis larvae, were 85 pg (3.57 ng a.i./g body weight) in dead larvae, and 82 pg (3.42 ng a.i./g body weight) in alive individuals. After topical treatment of H. didymator cocoons, most of the compound was retained in the silken cocoon, preventing contamination of the pupa. Also in the parasitoid adults, relatively low amounts of spinosad were accumulated in the body overall, but half of all the insecticide recovered was found in the ovaries. The kinetic results obtained help to better understand the toxicity of spinosad in the complex S. littoralis–H. didymator, and to ascertain the compatibility between spinosad and the parasitoid for optimizing the control of lepidopteran pests.     

Semi-field assessment of Spinosad in combination with Altosid briquet and Dudim DT tablets against Aedes aegypti mosquito larvae reared in pond water

This study aims at evaluating the larvicidal efficacy of slow-release formulations of bacterial insecticide Spinosad blended with two insect growth regulators (IGRs), Altosid XR–briquets and Dudim DT tablets against mosquito larvae of Aedes aegypti. This insect is the cause of dengue arthropod-borne viral disease. Treatment based on vector control using chemicals poses risks to human and environment, while the strategy based on the use of natural products poses lower risks. The results of the present study indicated that the mixture of Spinosad plus Dudim or Altosid tablets continued their fatal activity against the 3rd instar larvae for a longer period than the treatment with Spinosad, Altosid or Dudim tablets alone. The treatment of Spinosad plus either Altosid or Dudim has given effect to female mosquitoes in laying eggs in egg cups containing the treated pond water compared to the control egg cups. The results also showed that the treatment of Spinosad plus Altosid did not affect the egg hatching rate in the treated pond water, while the treatment of Spinosad combination with Dudim showed a significant decrease in the percentage of egg hatching. Use of natural products with larvicidal activities offers better approaches to integrated pest management and insecticide resistance management.