Biological control of the diamondback moth, Plutella xylostella: A review

The diamondback moth (DBM), Plutella xylostella (L.) (Lepidoptera: Plutellidae), is one of the most destructive cosmopolitan insect pests of brassicaceous crops. It was the first crop insect reported to be resistant to DDT and now, in many crucifer producing regions, it has shown significant resistance to almost every synthetic insecticide applied in the field. In certain parts of the world, economical production of crucifers has become almost impossible due to insecticidal control failures. Consequently, increased efforts worldwide have been undertaken to develop integrated pest management (IPM) programs, principally based on manipulation of its natural enemies. Although over 130 parasitoid species are known to attack various life stages of DBM, most control worldwide is achieved by relatively few hymenopteran species belonging to the ichneumonid genera Diadegma and Diadromus, the braconid genera Microplitis and Cotesia, and the eulophid genus Oomyzus. DBM populations native to different regions have genetic and biological differences, and specific parasitoid strains may be associated with the specific DBM strains. Therefore, accurate identification based on genetic studies of both host and parasitoid is of crucial importance to attaining successful control of DBM through inoculative or inundative releases. Although parasitoids of DBM larvae and pupae are currently its principal regulators, bacteria-derived products (e.g., crystal toxins from Bacillus thuringiensis) and myco-insecticides principally based on Zoophthora radicans and Beauveria bassiana are increasingly being applied or investigated for biological control. Viruses, nematodes and microsporidia also have potential as biopesticides for DBM. When an insect pest is exposed to more than one mortality factor, there is the possibility of interactions that can enhance, limit, or limit and enhance the various aspects of effectiveness of a particular control tactic. This paper reviews the effectiveness of various parasitoids and entomopathogens against DBM, interactions among them, and their possible integration into modern IPM programs.     

Advances in Transgenic Research for Insect Resistance in Sugarcane

The first phase of transgenic research in sugarcane concentrated on the development and evaluation of transgenic lines transformed for resistance to biotic stresses, particularly diseases and insect pests. Sugarcane is attacked by a range of insects including tissue borers, sucking pests and canegrubs. Losses due to these pests are estimated to be around 10%. Although chemical control and integrated pest management are regularly practiced for the control of insect pests, success is often limited due to practical difficulties. The genetic complexity of sugarcane coupled with the non-availability of resistance genes in the germplasm has made conventional breeding for insect resistance difficult. In this context, transgenic technology has become a handy tool for imparting insect resistance to an elite variety which is otherwise superior for most other agronomic traits. A number of transgenic sugarcane lines have been developed with genes expressing Cry proteins, proteinase inhibitors or lectins resistant to borers, sucking insects or grubs. While commercializing transgenic lines, issues such as higher and stable transgene expression, preparedness for resistance management and non-target effects need to be addressed. To manage the constant threat of resistance development in target insects, it is imperative to deploy field-level strategies taking clues from other crops coupled with the search for new potent replacement molecules for transformation.     

Tebufenozide resistance is associated with sex‐linked inheritance in Plutella xylostella

The diamondback moth (DBM), Plutella xylostella (L.), is a major pest of cruciferous crops. Tebufenozide, a novel nonsteroidal ecdysone agonist, exhibits good efficacy and has played an increasingly important role in the control of Lepidopteran pests in China. For its resistance management, the genetic basis of tebufenozide resistance was studied using a laboratory selected resistant strain of DBM (resistant ratio, RR = 268). A series of crosses with laboratory susceptible and resistant strains revealed that tebufenozide resistance in this pest was partially biased toward female heredity, with a large difference in RR for F₁ (RR = 29) and rF₁ progeny (RR = 147). The dominance calculated for these 2 cross progeny was ?0.788 and 0.09, respectively. Further analysis showed that the susceptible male and female larvae were similar in their sensitivity to tebufenozide, but the resistant female larvae showed significantly higher resistance than the resistant male larvae. The heredity of tebufenozide resistance in DBM might be linked with the W sex chromosome, which suggested that DBM has the ability to develop high levels of resistance to tebufenozide. This is the first report of sex‐linked inheritance of tebufenozide resistance in P. xylostella (L.).     

Behavioral Avoidance - Will Physiological Insecticide Resistance Level of Insect Strains Affect Their Oviposition and Movement Responses?

Agricultural organisms, such as insect herbivores, provide unique opportunities for studies of adaptive evolutionary processes, including effects of insecticides on movement and oviposition behavior. In this study, Brassica leaves were treated with one of two non-systemic insecticides and exposed to two individual strains (referred to as single or double resistance) of diamondback moth (Plutella xylostella) (DBM) exhibiting physiological resistance. Behavioral responses by these two strains were compared as part of characterizing the relative effect of levels of physiological resistance on the likelihood of insects showing signs of behavioral avoidance. For each DBM strain, we used choice bioassays to quantify two possible types of behavioral avoidance: 1) females ovipositing predominantly on leaf surfaces without insecticides, and 2) larvae avoiding insecticide-treated leaf surfaces. In three-choice bioassays (leaves with no pesticide, 50% coverage with pesticide, or 100% coverage with pesticide), females from the single resistance DBM strain laid significantly more eggs on water treated leaves compared to leaves with 100% insecticide coverage (both gamma-cyhalothrin and spinetoram). Females from the double resistance DBM strain also laid significantly more eggs on water treated leaves compared to leaves with 100% gamma-cyhalothrin, while moths did not adjust their oviposition behavior in response to spinetoram. Larvae from the single resistance DBM strain showed a significant increase in mobility in response to both insecticides and avoided insecticide-treated portions of leaves when given a choice. On the other hand, DBM larvae from the double resistance strain showed a significant decrease in mobility in response to insecticides, and they did not avoid insecticide-treated portions of leaves when given a choice. Our results suggest that pest populations with physiological resistance may show behavioral avoidance, as resistant females avoided oviposition on leaves without gamma-cyhalothrin. Thus, physiological resistance and behavioral avoidance do not appear to be controlled by the same selection pressures, and the mechanisms responsible for behavioral avoidance may vary among life stages. Our analysis also suggested that a population with lesser physiological resistance to insecticides may be under a stronger selection pressure and therefore be more likely to develop avoidance behaviors than a population with higher levels of physiological resistance.     

Use of cowpea trypsin inhibitor (CpTI) to protect plants against insect predation

Transgenic tobacco plants expressing a cowpea trypsin inhibitor gene have enhanced levels of insect resistance to a variety of insect pests. Furthermore, insect bioassay has shown the cowpea trypsin inhibitor to have anti-metabolic activity to insect pests of the orders Lepidoptera, Coleoptera and Orthoptera. The advantages and disadvantages of this approach to developing insect resistant crops is discussed in relationship to other methods, including conventional plant breeding and chemical control. Eventually it is hoped that African farmers will benefit from this industrially sponsored research.     

华北棉区主要害虫抗药性监测与治理技术示范

【目的】华北地区转基因Bt棉大面积种植后,刺吸式口器害虫已成为棉花主要害虫。本研究旨在监测明确棉花主要害虫对田间常用杀虫剂抗性水平变化,以指导田间合理用药。【方法】2013-2015年分别采用叶片药膜法和点滴法系统监测了河北省邱县、山东省滨州市、河南省西华县棉铃虫Helicoverpa armigera、棉蚜Aphis gossypii、绿盲蝽Apolygus lucorum对常用杀虫药剂的抗性动态变化情况。【结果】棉铃虫对辛硫磷、高效氯氟氰菊酯抗性呈上升趋势,抗性倍数都在20倍以上,对甲氨基阿维菌素苯甲酸盐从敏感状态转为中等水平抗性,抗性倍数在10倍以上。棉蚜对杀虫剂抗性水平都比较高,特别是对氧化乐果、高效氯氰菊酯、吡虫啉都已产生了高水平抗性,抗性倍数都在100倍以上。绿盲蝽对吡虫啉从敏感状态转为中等水平抗性,抗性倍数在10倍以上,对马拉硫磷、灭多威等药剂抗性倍数还维持在10倍以下,对高效氯氟氰菊酯只监测到滨州种群产生了中等水平的抗性,抗性倍数达到了95倍。在明确棉花害虫抗药性水平的基础上,对山东滨州棉花害虫实施了以轮换用药为主的抗性治理示范,示范区比农户自防对照区减少3次用药,增加棉花产量7.53%,节本增收109.16元,取得了较好的示范效果和经济效益。【结论】当前华北棉区主要害虫抗性水平上升,急需开展以轮换用药为主的抗性治理措施。     

两种赤眼蜂对小菜蛾卵的寄生能力和种间竞争

在实验室内研究了本地天敌拟澳洲赤眼蜂和外来天敌短管赤眼蜂在不同卵龄和不同空间条件下对小菜蛾卵的寄生和种间竞争.结果表明,在0～62 h的小菜蛾卵上,拟澳洲赤眼蜂和短管赤眼蜂单独接蜂时对小菜蛾卵的寄生率、子代羽化率、子代雌蜂百分率随卵龄下降,而后代发育死亡率则随卵龄增加.短管赤眼蜂对各龄小菜蛾卵的寄生率、子代羽化率、子代雌蜂百分率均较拟澳洲赤眼蜂高,而子代蜂的发育死亡率则较拟澳洲赤眼蜂低.两种蜂对小菜蛾卵龄的要求都不太严格,在小于48 h的小菜蛾卵上都能产卵寄生,且寄生率高于50%,但短管赤眼蜂寄生对小菜蛾卵龄的要求更宽松.在两种蜂混合接蜂时,后代中短管赤眼蜂的比例在各处理中都高于50%,且随寄主卵龄增加,当小菜蛾卵龄大于48 h后,短管赤眼蜂的比例高达100%,说明短管赤眼蜂对小菜蛾有更强的寄生能力和竞争能力.拟澳洲赤眼蜂和短管赤眼蜂单独接蜂时,在4～22cm^3空间范围内,各处理间对小菜蛾卵的寄生率没有显著差异,而短管赤眼蜂的寄生率(76.4%～86%)略低于拟澳洲赤眼蜂(88.7%～92.3%).当空间大于53 cm^3时,寄生率显著下降,在102cm^3空间时显著降低到50%.混合接蜂时各处理间寄生率差异不显著.两种接蜂方式对后代羽化率和雌蜂百分率没有太大影响,蜂的后代发育死亡率在两种蜂单独接蜂时随空间而增加.在4～102cm^3空间范围内,混合接蜂后代雄蜂中短管赤眼蜂从80%以上降低到20%以下,说明短管赤眼蜂的竞争能力随接蜂空间的加大而降低.     

Transgenic broccoli with high levels of Bacillus thuringiensis Cry1C protein control diamondback moth larvae resistant to Cry1A or Cry1C

A synthetic Bacillus thuringiensis (Bt) cry1C gene was introduced into broccoli (Brassica oleracea ssp. italica) by Agrobacterium-mediated transformation. Twenty-one Cry1C transgenic plants were regenerated from 400 hypocotyl and petiole explants. Variable amounts of stable steady- state cry1C mRNA accumulated in different transgenic plants. Cry1C protein (up to 0.4% of total soluble protein) was produced in correlation with the cry1C mRNA levels. Leaf section and whole-plant bioassays were done using diamondback moth (DBM) larvae from lines susceptible to Bt or resistant to Cry1A or Cry1C proteins (Cry1AR or Cry1CR, respectively). Plants with high levels of Cry1C protein caused rapid and complete mortality of all three types of DBM larvae with no defoliation. Plants with lower levels of Cry1C protein showed an increasing differential between control of susceptible of Cry1AR DBM. This study demonstrated that high production of Cry1C protein can protect transgenic broccoli not only from susceptible or Cry1AR DBM larvae but also from DBM selected for moderate levels of resistance of Cry1C. The Cry1C- transgenic broccoli were also resistant to two other lepidopteran pests of crucifers (cabbage looper and imported cabbage worm). These plants will be useful in studies of resistance management strategies involving multiple transgenes. This revised version was published online in June 2006 with corrections to the Cover Date.     

A hybrid Bacillus thuringiensis delta-endotoxin gives resistance against a coleopteran and a lepidopteran pest in transgenic potato

Expression of Bacillus thuringiensis delta-endotoxins has proven to be a successful strategy for obtaining insect resistance in transgenic plants. Drawbacks of expression of a single resistance gene are the limited target spectrum and the potential for rapid adaptation of the pest. Hybrid toxins with a wider target spectrum in combination with existing toxins may be used as tool to mitigate these problems. In this study, Desiree potato plants were genetically modified to resist attack by insect species belonging to the orders Coleoptera and Lepidoptera, through the insertion of such a hybrid gene, SN19. Transgenic plants were shown to be resistant against Colorado potato beetle larvae and adults, potato tuber moth larvae, and European corn borer larvae. These are the first transgenic plants resistant to pests belonging to two different insect orders. In addition, the target receptor recognition of this hybrid protein is expected to be different from Cry proteins currently in use for these pests. This makes it a useful tool for resistance management strategies.     

Detection of parasitism in diamondback moth, Plutella xylostella (L.), using differential melanization and coagulation reactions

Diamondback moth (DBM), Plutella xylostella, is known for causing damage to Brassica crops and developing resistance to chemical and biological pesticides; it has become one of the most difficult pests to manage in many regions around the world. The only way to reduce reliance on pesticides is to maximize the role of natural control agents for integrated pest management programs and be able to incorporate the mortality from control agents into pest control decision-making. More than 90 hymenopterous parasitoids are associated with DBM worldwide; among them, Diadegma semiclausum, is a major endoparasitoid of P. xylostella. To optimize parasitism of pests in pest control decision-making, it is necessary to develop rapid and simple methods for distinguishing parasitized from non-parasitized larvae in the field. Here we report on a number of diagnostic tools to identify parasitized larvae. One is based on differential melanization reactions in hemolymph due to immune suppression in parasitized larvae. The lack of coagulation reactions in hemolymph provides a simple initial test, where squashing a non-parasitized larva onto nitrocellulose membrane traps chlorophyll-containing gut content on the membrane leaving a green dot of clotted gut material. However, in immune-suppressed parasitized larvae, the gut content was washed away in absence of coagulation reactions and the membrane lacks a green dot. This tool alone or combined with others, allows quick detection of parasitized caterpillars in the field. We further showed that the antibody MAb 9A5 can be used to detect D. semiclausum parasitized larvae of DBM in Western blots.     

转抗虫基因植物生态安全性研究进展

转抗虫基因植物如Bt棉花等已在美国、中国和澳大利亚等国家大规模商业化种植 ,有关转抗虫基因植物潜在的生态风险已引起广泛的关注。该文综述了转抗虫基因植物研究应用现状与安全性研究进展。主要内容包括 :转抗虫基因植物的种类及其对靶标害虫的抗性 ,对非靶标害虫和天敌发生的影响 ,对农田生态系统生物多样性的影响 ,靶标昆虫的抗性治理及转抗虫基因植物的基因漂移等     

Sustained susceptibility of pink bollworm to Bt cotton in the United States

Evolution of resistance by pests can reduce the benefits of transgenic crops that produce toxins from Bacillus thuringiensis (Bt) for insect control. One of the world's most important cotton pests, pink bollworm (Pectinophora gossypiella), has been targeted for control by transgenic cotton producing Bt toxin Cry1Ac in several countries for more than a decade. In China, the frequency of resistance to Cry1Ac has increased, but control failures have not been reported. In western India, pink bollworm resistance to Cry1Ac has caused widespread control failures of Bt cotton. By contrast, in the state of Arizona in the southwestern United States, monitoring data from bioassays and DNA screening demonstrate sustained susceptibility to Cry1Ac for 16 y. From 1996-2005, the main factors that delayed resistance in Arizona appear to be abundant refuges of non-Bt cotton, recessive inheritance of resistance, fitness costs associated with resistance and incomplete resistance. From 2006-2011, refuge abundance was greatly reduced in Arizona, while mass releases of sterile pink bollworm moths were made to delay resistance as part of a multi-tactic eradication program. Sustained susceptibility of pink bollworm to Bt cotton in Arizona has provided a cornerstone for the pink bollworm eradication program and for integrated pest management in cotton. Reduced insecticide use against pink bollworm and other cotton pests has yielded economic benefits for growers, as well as broad environmental and health benefits. We encourage increased efforts to combine Bt crops with other tactics in integrated pest management programs.     

Current status and future perspectives on natural enemies for pest control in Taiwan

ABSTRACT

In Taiwan, the agricultural policy, ‘Reduce the consumption of pesticide to half in the next 10 years’, was launched in 2017. Pesticide application, which results in contamination of food by chemical residues, pest resistance, and other adverse ecological effects, is a growing public and environmental concern. Pest control by natural predators is, thus, the best alternative. Biological control methods implemented based on insights obtained from studies on pest behaviour, rearing, and various crop management modes, increase the possibility of controlling pests in modern organic agricultural systems. More than a decade has passed since the first introduction of a predatory insect in Taiwan for pest control (in the 1990s). Predatory and parasitic natural enemies, including lacewing, predatory stink bugs, Orius, and parasitic wasps, were initially used for controlling thrips, aphids, spider mites, whiteflies, and lepidopteran pests. At present, there exists a wide range of integrated pest management (IPM) methods incorporating other non-chemical, biological, and agricultural methods. However, recently, there has been an increase in research and development on the utilisation of natural enemies of insects and the associated food safety issues. Mass production and release, storage, and handling techniques of insect predators and parasitoids have been successful in recent years. The final goal of present day research is to develop natural enemy products and provide an IPM-based model to farmers for using natural enemies in agricultural production systems, thereby reducing pesticide application and ensuring food security.     

DNA Sequencing Reveals the Midgut Microbiota of Diamondback Moth,Plutella xylostella (L.) and a Possible Relationship with Insecticide Resistance

Background

Insect midgut microbiota is important in host nutrition, development and immune response. Recent studies indicate possible links between insect gut microbiota and resistance to biological and chemical toxins. Studies of this phenomenon and symbionts in general have been hampered by difficulties in culture-based approach. In the present study, DNA sequencing was used to examine the midgut microbiota of diamondback moth (DBM), Plutella xylostella (L.), a destructive pest that attacks cruciferous crops worldwide. Its ability to develop resistance to many types of synthetic insecticide and even Bacillus thuringiensis toxins makes it an important species to study.

Methodology/Principal Findings

Bacteria of the DBM larval midgut in a susceptible and two insecticide (chlorpyrifos and fipronil) resistant lines were examined by Illumina sequencing sampled from an insect generation that was not exposed to insecticide. This revealed that more than 97% of the bacteria were from three orders: Enterobacteriales, Vibrionales and Lactobacillales. Both insecticide-resistant lines had more Lactobacillales and the much scarcer taxa Pseudomonadales and Xanthomonadales with fewer Enterobacteriales compared with the susceptible strain. Consistent with this, a second study observed an increase in the proportion of Lactobacillales in the midgut of DBM individuals from a generation treated with insecticides.

Conclusions/Significance

This is the first report of high-throughput DNA sequencing of the entire microbiota of DBM. It reveals differences related to inter- and intra-generational exposure to insecticides. Differences in the midgut microbiota among susceptible and insecticide-resistant lines are independent of insecticide exposure in the sampled generations. While this is consistent with the hypothesis that Lactobacillales or other scarcer taxa play a role in conferring DBM insecticide resistance, further studies are necessary to rule out other possibilities. Findings constitute the basis for future molecular work on the functions of insect midgut microbiota taxa and their possible role in conferring host resistance to toxins.     

The evolutionary origins of pesticide resistance

Durable crop protection is an essential component of current and future food security. However, the effectiveness of pesticides is threatened by the evolution of resistant pathogens, weeds and insect pests. Pesticides are mostly novel synthetic compounds, and yet target species are often able to evolve resistance soon after a new compound is introduced. Therefore, pesticide resistance provides an interesting case of rapid evolution under strong selective pressures, which can be used to address fundamental questions concerning the evolutionary origins of adaptations to novel conditions. We ask: (i) whether this adaptive potential originates mainly from de novo mutations or from standing variation; (ii) which pre‐existing traits could form the basis of resistance adaptations; and (iii) whether recurrence of resistance mechanisms among species results from interbreeding and horizontal gene transfer or from independent parallel evolution. We compare and contrast the three major pesticide groups: insecticides, herbicides and fungicides. Whilst resistance to these three agrochemical classes is to some extent united by the common evolutionary forces at play, there are also important differences. Fungicide resistance appears to evolve, in most cases, by de novo point mutations in the target‐site encoding genes; herbicide resistance often evolves through selection of polygenic metabolic resistance from standing variation; and insecticide resistance evolves through a combination of standing variation and de novo mutations in the target site or major metabolic resistance genes. This has practical implications for resistance risk assessment and management, and lessons learnt from pesticide resistance should be applied in the deployment of novel, non‐chemical pest‐control methods.     

Effective control of yellow stem borer and rice leaf folder in transgenic rice indica varieties Basmati 370 and M 7 using the novel δ-endotoxin cry2A Bacillus thuringiensis gene

Transgenic rice indica varieties Basmati 370 and M 7 expressing the novel cry2A (Bt) insecticidal gene were generated by particle bombardment. Molecular and biochemical analyses in R0 and R1 populations confirmed stable integration and expression of this novel Bt transgene. We estimated that the gene product was expressed up to 5% of total leaf protein. Insect feeding bioassays demonstrated that the Cry2A protein was effective against the yellow stem borer and the rice leaf folder, two major rice pests in the Indian Subcontinent. This is the first report of the control of major rice pests using this specific Bt gene. The cry2A gene can now be used in combination with other insecticidal genes for pyramiding resistance against insect pests. This will delay, or perhaps in combination with integrated pest management practices, prevent evolution of insect populations resistant to single insecticidal genes.     

Transgenic Plants for Insect Pest Control: A Forward Looking Scientific Perspective

One of the first successes of plant biotechnology has been the creation and commercialisation of transgenic crops exhibiting resistance to major insect pests. First generation products encompassed plants with single insecticidal Bt genes with resistance against major pests of corn and cotton. Modelling studies predicted that usefulness of these resistant plants would be short-lived, as a result of the ability of insects to develop resistance against single insecticidal gene products. However, despite such dire predictions no such collapse has taken place and the acreage of transgenic insect resistance crops has been increasing at a steady rate over the 9 years since the deployment of the first transgenic insect resistant plant. However, in order to assure durability and sustainability of resistance, novel strategies have been contemplated and are being developed. This perspective addresses a number of potentially useful strategies to assure the longevity of second and third generation insect resistant plants.     

害虫及害螨对阿维菌素抗药性研究进展

阿维菌素(avermectins)是一类新型高效广谱的生物源农药,对多种害虫及害螨具有极好的防效。随着阿维菌素在害虫及害螨防治中的广泛应用,害虫和害螨对其的抗性问题日益受到关注。文章综述国内外的最新研究结果表明:小菜蛾Pluttella xylostella(L.)、二斑叶螨Tetranychus urticate Koch等已对阿维菌素产生抗性,对阿维菌素产生抗性的害虫和螨并不总是表现适合度劣势,且抗性一旦产生敏感性较难以恢复;抗性遗传多数由多基因、不完全隐性控制;抗性机理涉及多种因素。综合分析发现害虫和螨对阿维菌素存在较大的、潜在抗性风险。     

Efficacy of genetically modified Bt toxins against insects with different genetic mechanisms of resistance

Transgenic crops that produce Bacillus thuringiensis (Bt) toxins are grown widely for pest control, but insect adaptation can reduce their efficacy. The genetically modified Bt toxins Cry1AbMod and Cry1AcMod were designed to counter insect resistance to native Bt toxins Cry1Ab and Cry1Ac. Previous results suggested that the modified toxins would be effective only if resistance was linked with mutations in genes encoding toxin-binding cadherin proteins. Here we report evidence from five major crop pests refuting this hypothesis. Relative to native toxins, the potency of modified toxins was >350-fold higher against resistant strains of Plutella xylostella and Ostrinia nubilalis in which resistance was not linked with cadherin mutations. Conversely, the modified toxins provided little or no advantage against some resistant strains of three other pests with altered cadherin. Independent of the presence of cadherin mutations, the relative potency of the modified toxins was generally higher against the most resistant strains.     

Baseline responses of Alphitobius diaperinus (Coleoptera: Tenebrionidae) to spinosad, and susceptibility of broiler populations in Eastern and Southern Australia

Spinosad was proposed as a potential chemical for control of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in Australian broiler houses after the detection of strong cyfluthrin resistance in many beetle populations. In 2004-2006, spinosad susceptibility of 13 beetle populations from eastern and southern Australian broiler houses and a cyfluthrin/fenitrothion-resistant reference population was determined using topical application, and was compared with the susceptibility of an insecticide-susceptible reference population. Comparisons of dose-response curves and baseline data showed that all populations, including the insecticide-susceptible population, were roughly equivalent in their response to spinosad, indicating no preexisting spinosad resistance. Two field populations, including the resistant reference population, which had confirmed cyfluthrin/fenitrothion-resistance, showed no cross-resistance to spinosad. There was no significant correlation between beetle weight and LC9.9. A discriminating concentration of 3% spinosad was set to separate resistant and susceptible individuals. Considering the levels of spinosad resistance that have been recorded in other insect pests, the sustained future usefulness of spinosad as a broiler house treatment will rely on effective integrated beetle management programs combined with carefully planned chemical use strategies.