Chitin Synthesis Inhibitor Research Articles

Fitness Costs in Diamondback Moth Plutella xylostella (L.) (Lepidoptera: Plutellidae) Resistant to Lufenuron, A Chitin-Synthesis Inhibitor Insecticide

The diamondback moth (DBM), Plutella xylostella, is the main pest of Brassicas crops worldwide, and its recorded resistance to 101 active ingredients indicates it is difficult to control. The purpose of this study was to investigate the hypothesis that P. xylostella has fitness costs associated with its resistance to lufenuron, a chitin-synthesis inhibitor insecticide. Thus, concentration–mortality bioassays were performed for susceptible (REC-S), resistant (BZR-R) populations, their progenies F1 and F1′, and one established population without selection pressure (BZR-Rns) after four generations. A fertility life table was used to assess the biological performance of the REC-S and BZR-R. BZR-Rns of P. xylostella. The larval stage, longevity, and survival differed between populations. The reproductive rate (R0) was significantly lower in the F1 (♀R × ♂S) (28.19) and F1′ (♀S × ♂R) (34.06) progenies compared with their parents, but not with the relaxed BZR-Rns (39.39). The mean generation time (T), intrinsic rate of population growth (rm), and doubling time (DT) differed between REC-S and progenies, with fitness of 0.52 and 0.64 for F1 and F1′, respectively. Overall, the results suggest that the resistance of P. xylostella to lufenuron is stable and that low fitness costs appear to be associated with resistance to lufenuron, although heterozygotes showed lower fitness than their parents. Strategies such as preserving refuge areas, rotation of modes of action, etc., are essential for resistance management and prolonging the efficacy of control agents; this highlights the importance of integrated insecticide resistance management.

Role of chitin synthases CHS1 and CHS2 in biosynthesis of the cyst wall of Cryptocaryon irritans

Cryptocaryon irritans, a protozoan parasite that infects marine fish, is characterized by a complex life cycle that includes a cyst-forming reproductive phase. However, the composition of the cyst wall and mechanism of its formation remain unclear. In this study, we identified chitin as a key component of the cyst wall using calcofluor white and wheat germ agglutinin, with Fourier-transform infrared spectroscopy confirming its β-form structure. Two chitin synthase genes, CHS1 and CHS2, were identified as being expressed throughout the life cycle and show close phylogenetic relationships with chitin synthase from ciliates. Incubation with specific anti-CHS1 and -CHS2 antibodies significantly reduced both the thickness and chitin content of the cyst wall, highlighting the critical role of these enzymes in chitin biosynthesis. Treatment with benzoylureas, which inhibit chitin synthesis, caused thinning of the cyst wall and downregulation of CHS gene expression, resulting in an 84 % reduction in the hatching rate after treatment with 0.01 mM CuSO4 compared with control tomonts. Western blot analysis demonstrated that recombinant CHS proteins are immunogenic, and tomonts from CHS-immunized grouper exhibited reduced size. These findings bridge a crucial knowledge gap in understanding of the C. irritans cyst wall and highlight promising targets for infection prevention and control strategies.

How do termite baits work? implication of subterranean termite colony demography on the successful implementation of baits.

In 1995, the launch of the first commercial chitin synthesis inhibitor (CSI) bait led to the transformation of the subterranean termite control industry around the world. Their slow mode of action, which relies on both their ability to be transferred among nestmates and termite molting biology, has made them cost-effective solutions for subterranean termite colony elimination while minimizing the introduction of pesticides into the soil toward an environmentally sustainable strategy. However, despite successful commercial implementations, the acceptance of their use varies within the pest control industry around the world. Notably, the nuanced complexity of how CSI baits lead to colony elimination upon feeding by termite foragers has, in part, remained elusive for the past 3 decades, allowing for long-lasting misconceptions to persist. A recent series of studies has since provided complementary elements of understanding how CSI baits utilize termites' inherent colony demography, behavior, and physiology to trigger colony elimination after a characteristic succession of events within the colony collapse process. I here provide a synthetic overview of subterranean termite colony characteristics when exposed to CSI baits using Coptotermes (Wasmann) (Blattodea: Heterotermitidae) as a primary model system. The changes in colony demography through the colony collapse reflect how the mode of action of CSI baits makes them a prime solution for sustainable subterranean termite pest management. Following decades of innovation, ongoing interactions among termite researchers, bait product manufacturers, and pest management providers must continue to bring solutions to existing and emerging termite pest problems around the world.

Putative APSES family transcription factor mbp1 plays an essential role in regulating cell wall synthesis in the agaricomycete Pleurotus ostreatus

The clade A APSES family transcription factors (Mbp1, Swi4, and Swi6) contribute to cell wall synthesis regulation in fungi. Herein, evolutionary relationships among these proteins were clarified by phylogenetic analysis using various ascomycetes and basidiomycetes, and then the detailed function of Mbp1 in cell wall synthesis regulation was analyzed in Pleurotus ostreatus. Our phylogenetic analysis revealed that Mbp1 and Swi6 are widely conserved among various fungi, whereas Swi4 is a protein specific for Saccharomycotina. In P. ostreatus, two putative clade A APSES family transcription factors, protein ID 83192 and 134090, were found and identified as Mbp1 and Swi6, respectively. The mbp1 gene was then disrupted through homologous recombination using P. ostreatus 20b strain (Δku80) as a host to obtain mbp1 disruption strains (Δmbp1). Disruption of mbp1 significantly decreased the growth rate and shortened aerial hyphae, suggesting that Mbp1 is involved in mycelial growth, especially aerial hyphal growth. Furthermore, thinner cell walls, decreased relative percentage of β-glucan, and downregulation of all β-glucan synthase genes were observed in Δmbp1 strains. Therefore, Mbp1 plays an essential role in β-glucan synthesis regulation in P. ostreatus. Disruption of mbp1 also impacted the expression profiles of chitin synthase genes, septum formation, and sensitivity to a chitin synthesis inhibitor, suggesting that Mbp1 also regulates chitin synthesis. In conclusion, Mbp1 is responsible for normal mycelial growth and regulates β-glucan and chitin synthesis in P. ostreatus. To the best of our knowledge, this is the first report on the detailed function of Mbp1 in cell wall synthesis regulation in fungi.

Chlorbenzuron downregulated HcLCP-17 expression by depressing two 20E-responsive transcription factors Br-C and βFTZ-F1 in Hyphantria cunea (Lepidoptera: Erebidae) larvae.

Cuticular proteins (CPs) play essential roles in forming cuticular structures in insects. However, the specific functions and regulatory mechanisms of CPs remain largely unexplored. In this study, the Larval cuticular protein 17 (HcLCP-17) gene was identified from Hyphantria cunea, a highly destructive and polyphagous forest pest. To investigate the role of HcLCP-17 in cuticular function and transcriptional regulation mediated by 20E-responsive transcription factors (ERTFs), we employed RNA interference (RNAi) and yeast one-hybrid assay techniques. Additionally, we examined the molecular mechanism by which chlorbenzuron, a type of benzoylphenylurea (BPU) that functions as a chitin synthesis inhibitor (CSI), affects the 20E signaling pathway and ultimately regulates HcLCP-17 expression. HcLCP-17 encodes a polypeptide consisting of 393 amino acids, which includes a chitin-binding domain. Silencing HcLCP-17 resulted in a disturbance in the structural organization of the larval cuticle and a notable reduction in chitin levels. HcLCP-17 expression was controlled by the interaction between Broad-Complex (Br-C)and beta Fushi Tarazu Factor-1 (βFTZ-F1) with its promoter fragment. Furthermore, the inhibitory effect of chlorbenzuron on HcLCP-17 expression was found to be potentially mediated by Br-C and βFTZ-F1. The study presents a novel mode of action for the 20E signaling pathway in regulating the expression of CPs and reveals the potential mode-of-action of BPUs in insect cuticles. These findings provide a theoretical basis for future utilization of LCP-17 as a pesticide target making a significant contribution to the development of effective pest management strategies. © 2024 Society of Chemical Industry.

Effects of chronic oral exposure to insecticide teflubenzuron on the midgut of the honey bee Apis mellifera workers: histopathological insights into pesticide toxicity.

The honey bee Apis mellifera plays a significant role as a pollinator of native and cultivated plants, by increasing the productivity of several cultures, preserving the flora, and producing forest seeds. However, bee populations are declining worldwide, including A. mellifera, due to Colony Collapse Disorder, mainly resulting from the constant use of pesticides in the crops. Teflubenzuron is a physiological insecticide that belongs to the benzoylurea group, which inhibits chitin synthesis, the main component of the insect integument classified as safe for non-target insects, including bees. However, its effect on non-target organs of insects remains unknown. The midgut is the main organ of the digestive tract, which works in digestion and absorption and may be exposed to pesticides that contaminate food resources. The present work aimed to verify if the insecticide teflubenzuron is toxic and has histopathological effects on the midgut of A. mellifera adult workers. Workers exposed orally and chronically to the field-realistic concentration of teflubenzuron present 81.54% mortality. The epithelium of the midgut of these bees presents high vacuolization, spherocrystals, cell fragments released to the organ lumen, apocrine secretion, nuclear pyknosis, loss of cell-cell contact, and damage to regenerative cell nests and to the peritrophic matrix. These results indicate that the chitin synthesis-inhibiting insecticide teflubenzuron is toxic to A. mellifera after chronic oral exposure, at realistic field concentration, although it is classified as non-toxic to adult and non-target insects.

Comparative effect of biopesticides against the fall armyworm Spodoptera frugiperda (J.E. Smith)

Summary The current study evaluated the effect of microbial-derived insecticides (abamectin and spinosad), two microbials (Beauveria bassiana (Balsamo) Vuillemin, Bacillus thuringiensis Berliner), and three chitin synthesis inhibitors (CSIs) (chlorfluazuron, hexaflumuron and lufenuron) against Spodoptera frugiperda larvae. Spinosad and abamectin caused pronounced mortality against second larval instar of S. frugiperda using the leaf dipping method. Spinosad induced higher toxicity (LC50 = 4.01 mg/L) than abamectin (LC50 = 8.33 mg/L) one day after treatment. The treatments with B. bassiana and B. thuringiensis caused higher mortality of S. frugiperda larvae 7 days after treatment with LC50 values of 3.0 × 105 spores/ml and 8.2 × 106 cells/ml, respectively. In the case of the CSIs, hexaflumuron showed higher toxicity than chlorfluazuron and lufenuron with LC50 values of 0.01, 0.009 and 0.005 mg/L 3, 7 and 10 days after treatment, respectively.

Exposure to teflubenzuron reduces drought tolerance of collembolans

Chitin synthesis inhibitors (CSIs) are commonly used insecticides compromising cuticle formation and structure in arthropods. Arthropods rely on intact cuticles to maintain water balance and cellular homeostasis to survive in different weather conditions. We hypothesized that physiological impacts of CSIs may make arthropods more vulnerable to harsh environmental conditions, such as extreme heat, cold or drought. The aim of this study was to investigate if pre-exposure to teflubenzuron (a common CSI) would influence Folsomia candida's (Collembola: Isotomidae) sensitivity to natural stressors. Here, we exposed adult collembolans to teflubenzuron through food for two weeks, then survivors were immediately divided into three groups for subsequent acute heat, cold, and drought exposure. After acute exposure to these natural stressors, the collembolans were moved to optimal conditions for a one-week recovery period during which their survival, time to regain reproduction, and egg production were examined. We analyzed the interaction between effects of teflubenzuron and natural stressors using a multiplicative model. No interaction between effects of teflubenzuron and heat was observed in any test endpoints. A synergistic interaction between effects of teflubenzuron and cold was observed in the time to regain reproduction. Both survival and egg production, on the other hand, showed synergistic interaction between effects of teflubenzuron and drought, as well as a tendency for longer reproduction recovery times. Our results suggest that pre-exposure to teflubenzuron reduces drought tolerance in F. candida, while its impact on heat or cold tolerance is minor or absent. This study is among the first to explore the combined effects of CSI and natural stressors on soil arthropods, providing more insight on potential risks posed by such chemicals in the environment.

Quantitative structure-activity relationship of 2,6-dimethoxy-N-(3-(4-substituted phenyl)isoxazol-5-yl)benzamide for the inhibition of chitin synthesis.

Previously, we found that 5-(2,6-dimethoxybenzoylamino)-3-phenylisoxazoles (IOXs) inhibit chitin synthesis in the cultured integument of Chilo suppressalis. In this study, IOXs with various substituents at the para-position of the 3-phenyl ring were synthesized, and the concentrations required to inhibit chitin synthesis to 50% (IC50) were determined for all compounds. The introduction of halogens-such as F, Cl, and Br-and small alkyls-such as Me, Et, Pr, and n-Bu-at the 3-phenyl ring slightly enhanced the activity. However, the activity decreased drastically with the introduction of NO2, CF3, and t-Bu. The quantitative analysis of the substituent effect at the 3-phenyl ring on chitin-synthesis inhibition using the Hansch-Fujita method showed that the hydrophobic substituent with the optimum value was favored for the activity, but the bulky substituent in terms of E s was detrimental to the activity.

Dissecting the manipulation of lufenuron on chitin synthesis in Helicoverpa armigera

Lufenuron, a benzoylurea chitin synthesis inhibitor, is effective against many insect pests. However, the insecticidal activity of lufenuron has not been completely elucidated, nor has its disturbing effect on chitin synthesis genes. In this study, bioassay results demonstrated an outstanding toxicity of lufenuron against Helicoverpa armigera larvae. The treated larvae died from abortive molting and metamorphosis defects, and severe separation of epidermis and subcutaneous tissues was observed. Treatment of 3rd- and 4th-instar larvae with LC25 lufenuron significantly extended the duration of larval and pupal stage, reduced the rates of pupation and emergence, and adversely affected pupal weight. Besides, lufenuron can severely reduce chitin content in larval integument, and the lufenuron-treated larvae showed reduced trehalose content in their hemolymph. Further analysis using RNA sequencing revealed that five chitin synthesis genes were down-regulated, whereas the expressions of two chitin degradation genes were significantly enhanced. Knockdown of chitin synthase 1 (HaCHS1), uridine diphosphate-N-acetylglucosamine-pyrophosphorylase (HaUAP), phosphoacetyl glucosamine mutase (HaPGM), and glucosamine 6-phosphate N-acetyl-transferase (HaGNPAT) in H. armigera led to significant increase in larval susceptibilities to LC25 lufenuron by 75.48%, 65.00%, 68.42% and 28.00%, respectively. Our findings therefore revealed the adverse effects of sublethal doses of lufenuron on the development of H. armigera larvae, elucidated the perturbations on chitin metabolism, and proved that the combination of RNAi and lufenuron would improve the control effect of this pest.

Laboratory efficacy and performance of several type of chitin synthesis inhibitors (CSIs) towards three species of subterranean termite (Blattodea:Rhinotermitidae, Termitdae)

Laboratory efficacy and performance of several type of chitin synthesis inhibitors (CSIs) towards three species of subterranean termite (Blattodea:Rhinotermitidae, Termitdae)

Long term efficacy of three chitin synthesis inhibitors and oxymatrine as ecofriendly seed protectants against Callosobruchus maculatus and Callosobruchuschinensis

The residual efficacy of three chitin synthesis inhibitors (CSIs) (chlorfluazuron, hexaflumuron and lufenuron) and oxymatrine at an application rate of 5.0 mg/kg was evaluated against Callosobruchus maculatus and Callosobruchus chinensis monthly for 5 consecutive months (6 bioassays). In each bioassay, the mortality of two insect adults was counted after 7 days, progeny reduction and cowpea weight loss percentages were recorded after 45 days. Our results revealed that CSIs caused high level of adult mortality of both insects and oxymatrine caused a complete adult mortality (100%) for five months storage. The tested CSIs produced a significant progeny reduction of C. maculatus and C. chinensis but did not achieve a complete reduction of F1 progeny during all storage periods. A complete progeny suppression of two insects and full cowpea seed protection were observed in all oxymatrine treatments during the entire storage periods. A remarkable protection of cowpea seeds was also achieved in the treatments with CSIs particularly chlorfluazuron during the first 3 months. These findings indicate CSIs and oxymatrine could be implemented in integrated pest management programs of both insect species.

Impact of temperature and soybean genotypes on biology of tobacco caterpillar and its mortality assessment with chitin synthesis inhibitors under in vitro condition

Impact of temperature and soybean genotypes on biology of tobacco caterpillar and its mortality assessment with chitin synthesis inhibitors under in vitro condition

Effects of solid and aqueous dietary diflubenzuron ingestion on some biological parameters in synthetic pyrethroid-resistant German cockroach, Blattella germanica L. (Blattodea: Ectobiidae).

Cockroaches, widespread pests found in metropolitan areas, are known as vectors of various disease agents, including viruses, fungi and antibiotic-resistant bacteria, as well as causing allergies in humans. Insect growth regulators have been used in pest management for several decades. These insecticides disrupt insect development and reproduction. Chitin synthesis inhibitors interfere with chitin biosynthesis in insects, causing abortive moulting and mortality, as well as inhibiting egg fertility, and larval hatching in insects. In this research, we evaluated the various effects of diflubenzuron, a chitin synthesis inhibitor, on synthetic pyrethroid-resistant German cockroach (Blattella germanica L. Blattodea: Ectobiidae), including ootheca production, oothecal viability, ootheca incubation time, the number of nymphs emerging from the ootheca and survivorship of nymphs. The cockroaches were fed diets that contained diflubenzuron, which was added to solid bait (impregnated fish food) and ingestible aqueous bait (impregnated cotton). Three concentrations (0.5%, 1% and 2%) were used in the experiments. As a result, diflubenzuron treatment led to ootheca production ranging from 60% to 100%; statistically, no difference was found between the treatment and the control groups. The number of nymphs emerging from the first and second ootheca was reduced by 40%-100% in the diflubenzuron-treated groups compared with the control. Nymphs exposed to diflubenzuron-impregnated solid bait and ingestible aqueous bait experienced mortality exceeding 92.1% and 66.27% within 15 days, respectively. In conclusion, diflubenzuron is a potential insecticide for use in cockroach baits to control B. germanica, as it caused high nymphal and embryonic mortality in the synthetic pyrethroid-resistant population and decreased the number of nymphs emerging from the ootheca.

Symbiotic bacteria confer insecticide resistance by metabolizing buprofezin in the brown planthopper, Nilaparvata lugens (Stål)

Buprofezin, a chitin synthesis inhibitor, is widely used to control several economically important insect crop pests. However, the overuse of buprofezin has led to the evolution of resistance and exposed off-target organisms present in agri-environments to this compound. As many as six different strains of bacteria isolated from these environments have been shown to degrade buprofezin. However, whether insects can acquire these buprofezin-degrading bacteria from soil and enhance their own resistance to buprofezin remains unknown. Here we show that field strains of the brown planthopper, Nilaparvata lugens, have acquired a symbiotic bacteria, occurring naturally in soil and water, that provides them with resistance to buprofezin. We isolated a symbiotic bacterium, Serratia marcescens (Bup_Serratia), from buprofezin-resistant N. lugens and showed it has the capacity to degrade buprofezin. Buprofezin-susceptible N. lugens inoculated with Bup_Serratia became resistant to buprofezin, while antibiotic-treated N. lugens became susceptible to this insecticide, confirming the important role of Bup_Serratia in resistance. Sequencing of the Bup_Serratia genome identified a suite of candidate genes involved in the degradation of buprofezin, that were upregulated upon exposure to buprofezin. Our findings demonstrate that S. marcescens, an opportunistic pathogen of humans, can metabolize the insecticide buprofezin and form a mutualistic relationship with N. lugens to enhance host resistance to buprofezin. These results provide new insight into the mechanisms underlying insecticide resistance and the interactions between bacteria, insects and insecticides in the environment. From an applied perspective they also have implications for the control of highly damaging crop pests.

Dissipation kinetics and decontamination of novaluron residues in cowpea, eggplant, and okra

Novaluron, a chitin synthesis inhibitor that belongs to the benzoylphenyl urea insecticides, is used against pests of vegetables. The dissipation kinetics of novaluron in cowpea, eggplant, and okra was studied in a field trial to determine its safety for use. The fruit samples drawn periodically were extracted and cleaned up using a modified QuEChERS method, and analyzed by liquid chromatography and tandem mass detection. Dissipation of novaluron applied on to fields of cowpea, eggplant, and okra at 100 gram of active ingredient per hectare followed first order kinetics with waiting period of 3.4 − 3.6 days. Processing of novaluron-contaminated fruit with different household methods also reduced the levels of novaluron residues to various extents.

Buprofezin affects the molting process by regulating nuclear receptors SfHR3 and SfHR4 in Sogatella furcifera

Nuclear receptors play a crucial role in various signaling and metabolic pathways, such as insect molting and development. Buprofezin (2-tert-butylimino-3-isopropyl-5-phenyl-perhydro-1, 3, 5-thiadiazin-4-one), a chitin synthesis inhibitor, causes molting deformities and slow death in insects by inhibiting chitin synthesis and interfering with their metabolism. This study investigated whether buprofezin affects insect ecdysteroid signaling pathway. The treatment of buprofezin significantly suppressed the transcription levels of SfHR3 and SfHR4, two nuclear receptor genes, in third-instar nymphs of Sogatella furcifera. Meanwhile, the transcription levels of SfHR3 and SfHR4 in first-day fifth-instar nymphs were induced at 12 h after 20E treatment. In addition, the silencing of SfHR3 and SfHR4 genes in first-day fifth-instar nymphs caused severe developmental delay and molting failure, resulting in a significant reduction of survival rates at 7.36% and 2.99% on the eighth day, respectively. Further analysis showed that the silencing SfHR3 and SfHR4 significantly inhibited the transcription levels of chitin synthesis and degradation-related genes. These results indicate that buprofezin can inhibits chitin synthesis and degradation by suppressing the signal transduction of 20E through SfHR3 and SfHR4, leading to molting failure and death. This study not only expands our understanding of the molecular mechanism of buprofezin in pest control but also lays a foundation for developing new control strategies of RNAi by targeting SfHR3 and SfHR4.

Feeding cessation after feeding on 20-hydroxyecdysone in the Formosan subterranean termite.

To control subterranean termite pests, chitin synthesis inhibitor (CSI) baits have been widely applied. Despite CSI baits having low impacts on the environment, they require a lengthy time period to eliminate colonies. 20-hydroxyecdysone (20E) was proposed to speed up the baiting process as it showed faster mortality than CSI baits. However, the efficacy of 20E has previously not been tested at the colony level prior to applying in the field. We compared the effect of 20E, 20E + noviflumuron, noviflumuron and untreated control using colonies of Coptotermes formosanus. Our result revealed that both 20E and 20E + noviflumuron did not accelerate colony elimination and termite activity remained relatively stable during the observation periods. To determine the limited effects of 20E, we further investigated feeding duration and consumption amount of 20E with different concentrations (control, 100 and 1000 ppm) for 10 days. Termites ceased feeding after 1 day in 100 and 1000 ppm treatment and 100% mortality was observed within 10 days in 1000 ppm 20E, while mortality in the 100 ppm 20E treated group was much lower than that in the 1000 ppm group. Furthermore, no termites molted in the control and termites died from hyperecdysonism in 1000 ppm 20E treatment, whereas about 20% of termites molted in 100 ppm 20E. This study demonstrated that 20E may not be suitable as a sole active ingredient to accelerate elimination of a subterranean termite colony, while CSI baits and lower concentrations of 20E may reduce the lengthy time period in colony elimination. © 2023 Society of Chemical Industry.

Can low concentration flufenoxuron treatment increase the pathogenicity or production of nucleopolyhedrovirus occlusion bodies in Spodoptera exigua (Hübner) or Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae)?

Nucleopolyhedroviruses (Baculoviridae) are lethal pathogens that naturally infect lepidopteran larvae. The chitin synthesis inhibitor flufenoxuron has been reported to potentiate the insecticidal properties of nucleopolyhedrovirus occlusion bodies (OBs) by degrading the integrity of the peritrophic matrix in lepidopteran larvae. In the present study we examined the effect of flufenoxuron treatment on susceptibility of Spodoptera exigua (Hübner) and Spodoptera frugiperda (JE Smith) fifth instars to their homologous nucleopolyhedroviruses, SeMNPV and SfMNPV, respectively. Preliminary bioassays indicated that a 1 mg/l concentration of flufenoxuron resulted in 6.6 - 13.3% mortality of larvae, which could be used in combination with OB inoculum concentrations of 1.5 × 104 OBs/ml of SeMNPV OBs or 1.3 × 10⁶ OBs/ml of SfMNPV OBs. Treatment of larvae with flufenoxuron did not significantly affect virus-induced mortality of either S. exigua (16.7 - 17.2%) or S. frugiperda (28.9 - 34.4%) and had no significant effect on the mean yield of OBs from virus-killed insects of either species. The identity of both SeMNPV and SfMNPV in experimental insects was confirmed by examination of the restriction endonuclease profiles of each virus. These findings contrast with the results of previous studies in which high concentrations of flufenoxuron (100 mg/l) greatly potentiated virus-induced mortality in the silkworm Bombyx mori L., although insect mortality due to flufenoxuron treatment alone was not reported, for reasons that are unclear. We conclude that flufenoxuron (1 mg/l) failed to potentiate the insecticidal activity of SeMNPV or SfMNPV in their respective hosts.

Management of khapra beetle, Trogoderma granarium Everts, using binary combinations of chitin synthesis inhibitors and inert dusts

Control of Trogoderma granarium is difficult due to its high tolerance to all common insecticides. This problem can be partially overcome using binary combinations of insecticides with other control measures and products. In this study, three chitin synthesis inhibitors (CSIs), chlorfluazuron (CH), hexaflumuron (HE) and lufenuron (LU), and three inert dusts, diatomaceous earth (DE), zeolite (ZE) and kaolin (KA) and their combinations were assessed on stored wheat for the control of T. granarium. The three CSIs were tested at 2.5 and 5.0 mg/kg and the three inert dusts at 500 and 1000 mg/kg. The mortality of T. granarium larvae was counted after 7 and 14 days. The adult emergence (F1 progeny) from treated larvae was examined after 30 days, while F2 progeny and wheat weight loss were recorded after 80 days of exposure. The results showed that the treatments of wheat grains with individuals and combinations of CSIs and inert dusts caused remarkable mortality of T. granarium larvae after 14 days of exposure. Also, the combinations of CSIs and inert dusts were significantly more effective than those of products alone. The combinations (5.0 mg/kg of CH + 1000 mg/kg of DE and 5.0 mg/kg of LU + 1000 mg/kg of DE) induced a complete larval mortality after 14 days. Furthermore, individual and binary combinations decreased the adult emergence (F1 and F2 progeny) from T. granarium larvae and the complete inhibition of progeny was obtained in the treatments with CH or LU alone or with their combinations with the three inert dusts. After 80 days of exposure, all individual and binary treatments of CSIs and inert dusts particularly CH and LU, induced a complete wheat protection with weight loss of 0.0%. Our findings indicate that the binary combinations could be successfully applied in IPM strategies of this invasive insect species.