Lufenuron treatment temporarily represses gene expression and affects the SUMO pathway in liver of Atlantic salmon.

Lufenuron is a benzoylurea insecticide currently in use to combat sea lice infestation in salmon aquaculture in Chile. With pending approval in Norway, the aim of this work was to study the uptake and toxicity of lufenuron in liver tissue of Atlantic salmon. Juvenile salmon weighing 40 g were given a standard 7-day oral dose, and bioaccumulation and transcriptional responses in the liver were examined 1 day after the end-of-treatment (day 8) and after 1 week of elimination (day 14). Bioaccumulation levels of lufenuron were 29 ± 3 mg/kg at day 8 and 14 ± 1 mg/kg at day 14, indicating relatively rapid clearance. However, residues of lufenuron were still present in the liver after 513 days of depuration. The exposure gave a transient inhibition of transcription in the liver at day 8 (2437 significant DEGs, p-adj < .05), followed by a weaker compensatory response at day 14 (169 significant DEGs). Pathways associated with RNA metabolism such as the sumoylation pathway were most strongly affected at day 8, while the apelin pathway was most profoundly affected at day 14. In conclusion, this study shows that lufenuron easily bioaccumulates and that a standard 7-day oral dose induces a transient inhibition of transcription in liver of salmon.

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.

Sublethal and transgenerational effects of lufenuron on the biological traits of Panonychus citri (McGregor) (Acari: Tetranychidae).

The citrus red mite, Panonychus citri (McGregor), is a globally important pest that has developed severe resistance to various pesticides. Lufenuron has been widely used in the control of the related pests in citrus orchard ecosystem. In this study, the susceptibilities of egg, larva, deutonymph and female adult of P. citri to lufenuron was determined, and the LC50 values were 161.354 mg/L, 49.595 mg/L, 81.580 mg/L, and 147.006 mg/L, respectively. Life-table analysis indicated that the fecundities were significantly increased by 11.86% and 26.84% after the mites were treated with LC20 concentrations of lufenuron at the egg or deutonymph stages, respectively. After eggs were treated with lufenuron, the immature stage and longevity were also affected, and resulted in a significant increase in r, R0 and λ. After exposure of female adults to LC20 of lufenuron, the fecundity and longevity of F0 generation significantly decreased by 31.99% and 10.94%, respectively. Furthermore, the expression level of EcR and Vg was significantly inhibited upon mites was treated with lufenuron. However, lufenuron exposure has a positive effect on fecundity and R0 in F1 generation, the expression of all reproduction-related genes was significantly up-regulated. In conclusion, there was a stimulating effect on the offspring population. Our results will contribute to the assessment of the resurgence of P. citri in the field after the application of lufenuron and the development of integrated pest control strategies in citrus orchards.

Sublethal and transgenerational effects of lufenuron on biological characteristics and expression of reproductive related genes in the fall armyworm, Spodoptera frugiperda.

The fall armyworm, Spodoptera frugiperda, is a notorious polyphagous pest that causes serious economic losses in crucial crops and has invaded Africa and Asia. Lufenuron is widely used for controlling S. frugiperda in China, owing to its high toxicity against this key pest, and less pollution and little impact on natural enemies. In the present study, the sublethal and transgenerational effects of lufenuron on S. frugiperda were investigated to provide in-depth information for the rational use of lufenuron. Results showed that the development time and pupae weight were not significantly affected following exposure of females to LC10 and LC25 and male S. frugiperda to the LC10 of lufenuron. However, LC25 exposure significantly reduced pupal and total development time and pupae weight of male S. frugiperda. The longevity of S. frugiperda adults was prolonged by lufenuron and the fecundity of S. frugiperda treated with LC10 of lufenuron was significantly increased by 40% compared to the control. In addition, our study demonstrated that the LC25 of lufenuron had transgenerational effects on the progeny generation. The development time of female S. frugiperda whose parents were exposed to LC25 of lufenuron was significantly decreased compared to the control. And then, the expression profiles of Vg, VgR, JHEH, JHE, JHAMT, JHBP, CYP307A1, CYP306A1, CYP302A1 and CYP314A1 genes involved in insect reproduction and development were analyzed using Quantitative Real-Time PCR (RT-qPCR). Results showed that Vg, VgR, JHE, JHAMT, and CYP306A1 were significantly upregulated at the LC10 of lufenuron, which revealed that these upregulated genes might be linked with increased fecundity of S. frugiperda. Taken together, these findings highlighted the importance of sublethal and transgenerational effects under laboratory conditions and these effects may change the population dynamics in the field. Therefore, our study provided valuable information for promoting the rational use of lufenuron for controlling S. frugiperda.

Biological and physiological responses of two Bradysia pests, Bradysia odoriphaga and Bradysia difformis, to Dinotefuran and Lufenuron.

Bradysia odoriphaga and Bradysia difformis are destructive root maggots that cause severe losses to vegetables, flowers and edible fungi. Due to the long-term dependence on single pesticides, Bradysia resistance to insecticides has increased, and field control efficacy has decreased obviously. To screen alternative insecticides, and compare the insecticide susceptibility of these two species, we tested the toxicity of eight insecticides to B. odoriphaga and B. difformis, and measured the sublethal effects of Dinotefuran and Lufenuron on life-history parameters and detoxification enzyme activities. Bioassay results indicated that Dinotefuran and Lufenuron had relatively higher toxicity to B. odoriphaga and B. difformis compared to other neonicotinoid and insect growth regulator insecticides, respectively. Significant adverse impacts caused by sublethal concentrations (LC20) of Dinotefuran and Lufenuron on the life-history parameters of F0 and F1 generations of B. odoriphaga and B. difformis were observed. These included reduced survival, prolonged larval development and reduced adult longevity and fecundity. B. odoriphaga had greater resistance and adaptation to insecticides than B. difformis, and an LC20 concentration of Dinotefuran stimulated the reproduction of B. odoriphaga F1 generation and increased the life table parameters. Detoxifying enzymes (CarE and GSTs) and P450 activities fluctuated after a sublethal concentration (Dinotefuran and Lufenuron) treatment, and at the peak value of enzyme activities, the enhancement of detoxifying enzymes of B. odoriphaga was significantly higher than that of B. difformis. These results indicated that Dinotefuran and Lufenuron should be considered as alternatives to other insecticides for control of root maggots. B. odoriphaga exhibited stronger adaptation to insecticides than B. difformis. These data provide guidance for control of root maggots, and the basic information presented here can help reveal the differences in adaptive mechanisms between B. odoriphaga and B. difformis.

Toxic evaluation of Proclaim Fit® on adult and larval worker honey bees.

Impacts to honey bees due to exposure to agricultural pesticides is one of the most serious threats to the beekeeping industry. Our research evaluated toxicity of the formulated insecticides Lufenuron+Emamectin benzoate (Proclaim Fit®) on the European honey bee Apis mellifera L. at field-realistic concentration (worst-case scenario). Newly emerged (≤24-h old) and forager (unknown age) worker bees were treated with the field recommended concentration of Proclaim Fit® using three routes of exposure including residual contact, oral, and spray within the laboratory. We also assessed the effects of Proclaim Fit® on the specific activity of some well-known detoxifying enzymes including α-esterase, ß-esterase, and Glutathione S-transferase (GST) in the honey bees. In addition, toxicity of the formulation was tested on 4th instar larvae within the hive. Based on estimated median survival times (MSTs), Proclaim Fit® was highly toxic to the bees, especially when applied as spray. According to our estimated relative median potency (RMP) values, newly emerged bees were 1.72× more susceptible than foragers to Proclaim Fit® applied orally. Enzyme assays revealed the considerable involvement of the enzymes, especially GST and α-esterase, in detoxification of the Proclaim Fit®, but their activities were significantly influenced by route of exposure and age of bee. Notably, Proclaim Fit® was highly toxic to 4th instar honey bee larvae. Our results generally indicate a potent toxicity of Proclaim Fit® toward honey bees. Therefore, its application requires serious consideration and adherence to strict guidelines, especially during the flowering time of crops.

Ameliorative effect of avocado oil against lufenuron induced testicular damage and infertility in male rats.

Lufenuron is a benzoylurea pesticide that causes significant histological and histochemical damage in mammals. Avocado is a common food in the human diet that contains antioxidant and antitumor properties. In male rats, avocado oil's protection against lufenuron-induced reproductive deterioration, oxidative stress, and DNA damages was investigated. Twenty-eight mature male rats were selected and distributed into four groups: Group 1, control group were administered distilled water orally; Group 2 received 4 ml/kg avocado; Group 3 was given lufenuron (1.6 mg/kg), and Group 4 was given avocado oil/lufenuron. The findings show that lufenuron treatment reduces reproductive hormone levels, sperm count, motility, viability and causes negative histopathological changes in testicular tissue, such as decreased epithelial height and increased luminal diameter degenerated spermatogenesis. Furthermore, lufenuron reduced the content of antioxidant enzymes while increasing the level of malondialdehyde, nitric oxide and corresponding DNA damage. Results showed that lufenuron is associated with testicular function impairment, which leads to infertility. Treatment with avocado oil improved reproductive hormone secretions, enzymatic activity, histological and DNA damage parameters in testis tissues, reducing the negative effects of lufenuron, proving that it may have a therapeutic role against lufenuron-mediated testicular toxicity.

Knockdown of UDP-N-acetylglucosamine pyrophosphorylase and chitin synthase A increases the insecticidal efficiency of Lufenuron to Spodoptera exigua.

Spodoptera exigua (Lepidoptera, Noctuidae) has been responsible for causing considerable and widespread agricultural losses worldwide. Owing to strong selective pressure, S. exigua showed increased resistance to Lufenuron (LUF). Consequently, RNA interference (RNAi)-based insecticides had more benefits than chemical insecticides. Therefore, to enhance the insecticidal activity of LUF to S. exigua, in the present study, we aimed to elucidate the impact of double-stranded RNAs (dsRNAs) on S. exigua larval susceptibility to LUF. First, the transcriptome of S. exigua was sequenced following the treatment with LUF. By comparing the upregulated and downregulated GO enrichment, chitin binding and chitin metabolic processes were the significantly enriched pathways. According to transcriptome sequencing, 8 genes associated with chitin biosynthesis, 8 chitin degradation genes, and 17 cuticle protein genes were obtained. UDP-N-acetylglucosamine pyrophosphorylase (UAP) and Chitin synthase A (CHSA) showed significantly downregulated expression after treatment with different sublethal doses of LUF. Downregulation of UAP increased mortality from 31.97% to 47.91% when the larvae were exposed to LUF. A significant increase in the mortality of S. exigua from 30.63% to 50.19% was observed following LUF administration after dsCHSA. In addition, the expression analysis of genes associated with chitin biosynthesis was significantly changed after LUF treatment, dsRNAs-RNAi, and their combination (LUF-dsRNAs). Significant differences were observed in the chitin content between the control group at 72 h after treatments. Results of the present study can help further elucidate the understanding of the combined effects of RNAi and LUF on S. exigua. Additionally, this research provides a suitable foundation for future studies with the aim to develop an efficient method of delivery for large-scale pest control in the fields.

[Two case report of death from acute compound chlorfenapyr poisoning].

Chlorfenapyr is a moderately dangerous insecticide widely used in agriculture. The mortality of acute poisoning patients is high, and there is no effective treatment. This paper retrospectively analyzes the clinical data of two cases of compound chlorfenapyr poisoning. The main symptoms of the patients were high fever, sweating, gradual coma, increased creatine kinase and myoglobin, with delayed poisoning symptoms. Despite comprehensive treatment, both patients died eventually. It indicated that chlorfenapyr was highly toxic and had a high mortality. In addition to routine symptomatic treatment for patients with acute poisoning, blood purification treatment should be actively carried out in the early stage.

Residue behavior and dietary risk assessment of six pesticides in pak choi using QuEChERS method coupled with UPLC-MS/MS.

A reliable and simple modified QuEChERS method with UPLC-MS/MS was developed for the simultaneous determination of six pesticides (dimethomorph, imidaclothiz, lufenuron, methoxyfenozide, pyridaben, spinetoram) and their metabolites in pak choi. Method validation indicated good linearity (R2 ≥ 0.99), accuracy (recoveries of 75%-112%), sensitivity (limits of quantification, 0.002-0.01 mg kg-1), and precision (relative standard deviations ≤ 21%), and matrix effects were -36-28%. The half-lives of the six pesticides in pak choi were 2.2-12 d under open field and greenhouse conditions. Considering the short growth cycle of pak choi, the terminal residue levels (0.046-7.8 mg kg-1) and the relevant maximum residue limits (MRLs) of some countries, 5 d was recommended as the pre-harvest interval for the six pesticides on pak choi. Dietary risk assessment revealed that the risk quotients were 3.1%-58% for different gender and age groups in China, indicating none unacceptable public health risk for general population. The results showed that all the six pesticides degraded faster and the terminal residues were much lower under open field conditions than those under greenhouse conditions, which was mainly due to the influence of rainfall, sunlight and other environmental factors. This work was thus significant in assessing the dissipation fate and food safety risks of the six pesticides on pak choi and facilitated the establishment of maximum residue limits.

SAFETY AND EFFICACY OF MILBEMYCIN OXIME AND LUFENURON TO TREAT ARGULUS SPP. INFESTATION IN SMOOTH BACK RIVER STINGRAYS (POTAMOTRYGON ORBIGNYI) AND MAGDALENA RIVER STINGRAYS (POTAMOTRYGON MAGDALENAE).

Fish lice, ectoparasites of the genus Argulus, are branchurian crustaceans that can significantly impact fish health by causing mechanical damage to cutaneous barriers and increasing susceptibility to other infections. While many treatments have been reported in teleosts and invertebrates, there are no published treatments for elasmobranchs. This study evaluated the safety and efficacy of a commercial formulation of milbemycin oxime and lufenuron in freshwater stingrays for treatment of Argulus spp. Seven juvenile Magdalena river stingrays (Potamotrygon magdalenae) and 10 juvenile smooth back river stingrays (Potamotrygon orbignyi) had severe infestations of Argulus spp. that were identified visually and microscopically. Animals were treated with milbemycin oxime and lufenuron (at 0.015 mg/L and 0.30 mg/L, respectively) in a 6-hr immersion once weekly for two treatments. They were visually examined for skin lesions as well as behavior and appetite daily by animal care staff. A subset of animals was euthanized and necropsied on days 8, 9, 43, and 78 after treatment initiation. There were no Argulus spp. detected at the time of the second treatment. Complete gross and histologic evaluations were completed for all animals. At all time points, no gross abnormalities were detected with the exception of thin body condition in some animals; no Argulus spp. were noted. Histologic lesions were all attributed to poor nutritional state at the time of acquisition. No histologic evidence of acute or chronic toxicosis was detected. The commercial formulation of milbemycin oxime and lufenuron, applied at the dose and for the exposure time used in this study, effectively eradicated Argulus spp. in a population of juvenile P. magdalenae and P. orbignyi, and did not cause mortality or clinical gross or histologic evidence of acute or chronic toxicity.

Hematological, physiological and genotoxicological effects of Match 5% EC insecticide on Biomphalaria alexandrina snails.

Freshwater snails are used as brilliant biomarkers of aquatic ecosystem pollution by chemical compounds. The objective of this study is to highlight the ecotoxicological impacts of the insecticide Match 5%EC (its active ingredient is lufenuron 5% EC) on Biomphalaria alexandrina snails the intermediate host of Schistosoma mansoni in Egypt. The present investigation recorded a remarkable molluscicidal effect of lufenuron 5% EC on these snails and there was a decrease in total number of their hemocytes after exposure. Three morphologically distinct populations of circulating hemocytes were identified (round small cells, granulocytes and hyalinocytes) and results showed that some hyalinocytes had a shrunk nucleus and some were degenerated. Significant increase of transaminases (ALT and AST), while, a decrease of the total protein and albumin content in hemolymph was recorded. The results of alkaline comet assay in the present study demonstrated that lufenuron 5% EC has a genotoxic effect especially when its concentration increases. It can be concluded that Biomphalaria alexandrina snails can be used as bio monitor to screen the deleterious effects of lufenuron 5% EC insecticide as a cause of the environmental pollution, and this insecticide can be used in controlling schistosomiasis because of its molluscicidal effects on B. alexandrina snails.

Laboratory evaluation on a potential birth control diet for fruit fly sterile insect technique (SIT).

Sterile insect technique (SIT) is one of the most effective fruit fly control technologies. Irradiation has been used to sterilize male fruit flies before release to the field to compete with the wild males for females. Imagine an environmental and cost effective method using a rearing diet that can make insects sterile indefinitely, by feeding for 7days before release. This could replace costly irradiation process. A potential birth control diet was evaluated on fertility, mating, survival, and protein analysis for fruit fly species in Hawaii. Insects were continuously fed an agar diet with lufenuron (LFN) for 7d after emergence and then switched to a control diet to simulate the actual field condition. The influence on egg hatch was dose dependent. With dose of 2-4mg/g in the diet, egg hatch from LFN-fed was almost 100% suppressed for 24 experimental days if adults of Ceratitia capitate (Widemann), Bactrocera dorsalis (Hendel), and B. latifrons (Hendel) continued to feed on LFN diet. B. cucurbitae (Coquillett) was not affected by LFN. However, egg hatch from LFN fed B. latifrons and B. dorsalis were suppressed for at least 2weeks after switching to the control diet at 7d. Egg hatch did not recover >4% up to 24d. Proteome analysis revealed that ABD-4 protein was under expressed by 70-83% on LFN fed females and males of B. latifrons and B. dorsalis while Pbprp2 protein was significantly over expressed by 6-12 fold on LFN fed males only. These two proteins were not expressed in C. capitata and B. cucurbitae. Therefore, this report focused more on B. latifrons and B. dorsalis. This finding suggested a great potential for one alternative to sterilize fruit flies for SIT without irradiation.

Dissipation kinetics of emamectin benzoate and lufenuron residues in cabbage grown under field conditions.

Residue analysis of emamectin benzoate and lufenuron in cabbage matrices and soil was developed using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) method and ultra high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The samples were extracted with 1% acetic acid in acetonitrile (v/v) or 1% acetic acid in acetonitrile/water (5:1, v/v) and cleaned up by dispersive solid-phase extraction. Mean recoveries and relative standard deviations (RSDs) in all samples ranged 87.8-100.0 % and 3.6-12.6% for emamectin benzoate and 87.8-104.8 % and 6.2-11.5% for lufenuron, respectively. The validated method was used to evaluate the dissipation rate of emamectin benzoate and lufenuron in cabbage and soil as well as the residual levels in harvested cabbage and soil at different preharvest intervals (PHI). The half-lives of emamectin benzoate and lufenuron were 1.08-2.70 and 1.74-5.04 days in cabbage, and 1.42-4.01 and 0.94-6.18 days in soil, respectively. The terminal residues were below the China maximum residue limits (MRLs) at 3 days for emamectin benzoate (0.1 mg kg(-1)) and European Union MRLs at 5 days for lufenuron (0.5 mg kg(-1)), which suggested that 5 days could be recommended as the PHI for the commercial formulation of emamectin benzoate and lufenuron application in the Chinese cabbage field.

Dietary lufenuron reduces egg hatch and influences protein expression in the fruit fly Bactrocera latifrons (Hendel).

Lufenuron (LFN), a chitin synthase inhibitor, impacts the fertility of Ceratitis capitata, Bactrocera dorsalis, B. cucurbitae, and B. latifrons. We posed the hypothesis that LFN curtails egg hatch in the solanaceous fruit fly, B. latifrons. In this study, newly emerged virgin adults were sexed and fed for 12 days with varying concentrations of LFN-laced agar diets until sexual maturation. Eggs were collected from 12-d-old adults and the egg hatch was assessed. Egg hatch decreased in adults reared on LFN-treated diets. LFN-treated media did not influence fertility after one gender was reared on experimental and the other on control media before mating. Exposure to LFN-treated medium after mating led to reduced egg hatch. We infer that LFN is not a permanent sterilant, and reduced egg hatch depends on continuous exposure to dietary LFN after mating. Proteomic analysis identified two differentially expressed proteins, a pheromone binding protein and a chitin binding protein, between adults maintained on LFN-treated and control diets. Expression of two genes encoding chitin synthase 2, and chitin binding protein, was altered in adults exposed to dietary LFN. LFN treatments also led to increased expression of two odorant binding proteins one in females and one in males. We surmise these data support our hypothesis and provide insight into LFN actions.

Construction and biological effects of a redox-enzyme dual-responsive lufenuron nano-controlled release formulation.

BACKGROUND: Pesticide formulations based on nanotechnology can effectively improve the efficiency of pesticide utilization and reduce pesticide residues in the environment. In this study, mesoporous silica nanoparticles containing disulfide bonds were synthesized by the sol-gel method, carboxylated and adsorbed with lufenuron, and grafted with cellulose to obtain a lufenuron-loaded nano-controlled release formulation (Luf@MSNs-ss-cellulose). RESULTS: The structure and properties of Luf@MSNs-ss-cellulose were characterized. The results showed that Luf@MSNs-ss-cellulose exhibits a regular spherical shape with 12.41% pesticide loading. The highest cumulative release rate (73.46%) of this pesticide-loaded nanoparticle was observed at 7 days in the environment of glutathione and cellulase, which shows redox-enzyme dual-responsive performance. As a result of cellulose grafting, Luf@MSNs-ss-cellulose had a small contact angle and high adhesion work on corn leaves, indicating good wetting and adhesion properties. After 14 days of spraying with 20 mg L-1 formulations in the long-term control efficacy experiment, the mortality of Luf@MSNs-ss-cellulose against Ostrinia furnacalis larvae (56.67%) was significantly higher than that of commercial Luf@EW (36.67%). Luf@MSNs-ss-cellulose is safer for earthworms and L02 cells. CONCLUSION: The nano-controlled release formulation obtained in this study achieved intelligent pesticide delivery in time and space under the environmental stimulation of glutathione and cellulase, providing an effective method for the development of novel pesticide delivery systems. © 2023 Society of Chemical Industry.

Residue and dietary intake risk assessment of lufenuron and chlorfenapyr and its corresponding metabolite in cabbage under field conditions.

A simple, low-cost, and highly sensitive method using a modified QuECHERS procedure based on a liquid chromatography-tandem mass spectrometer (LC-MS/MS) was established to simultaneously quantify lufenuron and chlorfenapyr and the corresponding metabolite tralopyril in cabbage for the first time. On the basis of this method, terminal residue and dietary risk of lufenuron and chlorfenapyr in cabbage were investigated. The recoveries of lufenuron, chlorfenapyr, and tralopyril ranged from 88 to 110%, with relative standard deviation of less than 12.4%. The field trial results showed that at the pre-harvest interval (PHI) of 21 days, the terminal residues of lufenuron, chlorfenapyr, and tralopyril in the supervised trials were not higher than 0.02 mg/kg, and the highest detected residue levels of lufenuron, chlorfenapyr, and tralopyril were 0.047, 0.055, and <0.02 mg·kg-1 at 14-day pre-harvest respectively, which were lower than the maximum residue limits (MRLs) for cabbage established in China. For the dietary risk assessment, the national estimated daily intakes (NEDIs) as proportion of acceptable daily intakes (ADIs) were 80.4% and 29.9% for chlorfenapyr and lufenuron respectively indicating an acceptable dietary risk to Chinese population.

Identification and Functional Characterization of Two Chitin Synthases in the Black Cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae).

The black cutworm, Agrotis ipsilon (Hufnagel), a seasonal migrant and a prolific generalist, can feed on nearly all vegetables and grain crops, causing considerable economic impacts on a global scale. Given its cryptic nature, A. ipsilon management has been extremely challenging. Chitin synthase (CHS), a key enzyme involved in chitin biosynthetic pathway and crucially important for the growth and development of insects, is the molecular target of chitin synthesis inhibitors, a group of broad-spectrum insecticides that is compatible with Integrated Pest Management practices. In this study, we investigated the potential of targeting chitin synthases to control A. ipsilon. As a result, two chitin synthases, AiCHS1 and AiCHS2, were identified and cloned from A. ipsilon. The temporal-spatial distribution study showed that AiCHS1 was predominantly expressed at the pupal stage and most abundant among tissues of head capsule and integument, while AiCHS2 was mainly expressed at the sixth instar larval stage and tissues of foregut and midgut. RNAi-based functional study confirmed gene silencing caused significant reduction in the expression levels of the corresponding mRNA, as well as resulted in abnormal pupation and mortality, respectively. Furthermore, under the treatment of lufenuron, a chitin synthesis inhibitor, A. ipsilon responded with an elevated expression in AiCHS1 and AiCHS2, while larvae showed difficulty in shedding old cuticle, and a cumulative mortality of 69.24% at 48 h. In summary, chitin synthases are crucial for chitin biosynthesis in A. ipsilon and can be targeted for the control (e.g., RNAi-based biopesticides) of this devastating insect pest.

EVALUATING BAITS WITH LUFENURON AND NITENPYRAM FOR FLEA CONTROL ON PRAIRIE DOGS (CYNOMYS SPP.) TO MITIGATE PLAGUE.

Plague, caused by Yersinia pestis, is a widespread threat to endangered black-footed ferrets (Mustela nigripes) and their primary prey, prairie dogs (Cynomys spp.). Wildlife biologists most commonly manage plague using insecticides to control fleas, the primary vectors of Y. pestis. We tested edible baits containing the insecticides lufenuron and/or nitenpyram in prairie dogs. During a laboratory study, we treated 26 white-tailed prairie dogs (Cynomys leucurus) with lufenuron at 300 mg/kg body mass. All animals remained clinically healthy over the 9 wk monitoring period. Although serum lufenuron concentrations were >130 ppb in two treatment groups at week 1, concentrations declined to ≤60 ppb after 3 wk in non-torpid prairie dogs and after 7 wk in torpid prairie dogs. In a field experiment, we tested baits containing a combination of 75 mg lufenuron and 6 mg nitenpyram, respectively, in black-tailed prairie dogs (Cynomys ludovicianus). We uniformly distributed baits at 125 baits/ha on two plots (treated once) and 250 baits/ha on two plots (each treated twice 4.4 wk apart). Following treatments, flea abundance increased on prairie dogs and remained stable in burrows. Our findings indicate that baits containing lufenuron and nitenpyram, at the reported treatment rates, are ineffective tools for flea control on prairie dogs. Future experiments might evaluate efficacy of higher doses of lufenuron and nitenpyram, and repetitive treatments at differing intervals over time to evaluate potentially therapeutic treatments.

Effects of lufenuron treatments on the growth and development of Spodoptera frugiperda (Lepidoptera: Noctuidae).

Lufenuron is an effective benzoylurea insecticide that inhibits the synthesis of chitin and regulates the growth of insects. However, little is known about the effects of lufenuron treatment on the development of Spodoptera frugiperda (J. E. Smith). In this study, we assessed the toxicity of lufenuron on S. frugiperda and evaluated the effects of lufenuron treatment on the growth and development of S. frugiperda. The results showed that lufenuron exhibits high insecticidal activity against S. frugiperda, with the LC50 value of 0.99 mg L-1. Lufenuron treatments can significantly prolong the larval developmental duration and reduce the rates of pupation and emergence. To further explore the underlying mechanism of this observation, the expression profiles of the chitin synthase gene (SfCHS) and chitinase gene (SfCHT), two key enzyme genes involved in the molting of S. frugiperda, were determined after exposure to lufenuron for 96 h. The results of qRT-PCR demonstrated that lufenuron treatments can significantly reduce the expression of SfCHT, while the expression of SfCHS remained relatively stable. Furthermore, we found that lufenuron strongly interacted with chitinase (SfCHT) (-10.8 kcal/mol) and chitin synthase (SfCHS) (R1: -9.7 kcal/mol; R2: -10.2 kcal/mol). Our results indicated that lufenuron has significant effects on the development of S. frugiperda that might be attributed to the differential expression of SfCHT and SfCHS.