Piperonyl Butoxide Research Articles

Resistance of Anopheles gambiae s.s. against commonly used insecticides and implication of cytochrome P450 monooxygenase in resistance to pyrethroids in Lambaréné (Gabon)

BackgroundInsecticides are a crucial component of vector control. However, resistance constitute a threat on their efficacy and the gains obtained over the years through malaria vector control. In Gabon, little data on phenotypic insecticide resistance in Anopheles vectors are published, compromising the rational implementation of resistance management strategies. We assessed the susceptibility to pyrethroids, carbamates and organophosphates of Anopheles gambiae sensu lato (s.l.) and discuss the mechanisms involved in the pyrethroid resistance-phenotype.MethodsA. gambiae s.l. larvae were collected from breeding sites in Lambaréné. Emerging adults were used in WHO tube assays at an insecticide concentration that defines resistance (diagnostic concentration). Subsequently, deltamethrin and permethrin were used at 5x and 10x diagnostic concentrations and after preexposure with the cytochrome p450 (and glutathione S-transferase) inhibitor piperonyl butoxide (PBO). A subset of mosquitoes was typed by molecular methods and screened using Taqman assays for mutations conferring target site resistance at the Voltage-gated sodium channel 1014 (Vgsc-1014) locus and the acetylcholinesterase (Ace-1) gene.ResultsAll mosquitoes were A. gambiae sensu stricto (s.s.) and resistant to permethrin, deltamethrin and alphacypermethrin (mortality less than 98%). However, mosquitoes were susceptible to malathion but resistant to bendiocarb. The level of resistance was high for permethrin and at least moderate for deltamethrin. Pre-exposure to PBO significantly increased the mortality of resistant mosquitoes (P < 0.0001). They became fully susceptible to deltamethrin and permethrin-induced mortality increased 4-fold. The G119S Ace-1 resistance allele, which confers resistance to both organophosphates and carbamates, was not present. All sampled mosquitoes were either homozygous for the Vgsc-L1014F or heterozygous for Vgsc-L1014F/L1014S, a marker for resistance to pyrethroids and organochlorides.ConclusionThese findings demonstrate a role of cytochrome P450 monooxygenases in the pyrethroid-resistance of A. gambiae s.s. from Lambaréné. Combining PBO with pyrethroids, as done in second generation bednets, may be used to revert resistance. In addition, malathion could also be used in combination with pyrethroids-based methods for resistance management.

Genome-wide analysis of detoxification genes conferring diamide insecticide resistance in Spodoptera exigua identifies CYP9A40.

For over a decade, diamide insecticides have been effective against lepidopteran pests like beet armyworm, Spodoptera exigua (Hübner, 1808). However, the evolution of resistance poses a challenge to their sustainable use. We identified an I4790M mutation in the S. exigua ryanodine receptor (RyR) gene, but its correlation with resistance varied across the field-collected Korean populations of S. exigua. RNA sequencing and differential gene expression analysis were performed to investigate other resistance mechanisms. Diamide-resistant and susceptible strains and F1 hybrids were compared by mapping RNA-seq reads to the S. exigua reference genome. CYP9A40 was identified as a critical gene in diamide resistance due to its high expression in the resistant strains. Synergist bioassays with piperonyl butoxide supported the role of P450s in diamide metabolic resistance in S. exigua. A strong positive correlation between CYP9A40 over-expression levels (up to 80-fold) and diamide LC50 values was obtained for field-collected populations uniformly showing a 100% frequency of the RyR I4790M target-site resistance allele. To validate the function of CYP9A40 in diamide detoxification, we recombinantly expressed the gene and tested its ability to bind and degrade chlorantraniliprole as a substrate. The results confirmed its catalytic role in diamide metabolism. CYP9A40 has been identified and validated to confer metabolic resistance in Korean S. exigua populations. It works alongside the RyR target-site I4790M mutation to enhance diamide resistance. These mechanisms offer insights for resistance monitoring and support insecticide resistance management programs to improve control strategies for S. exigua.

An observational analysis of the impact of deltamethrin + piperonyl butoxide insecticide-treated nets on malaria case incidence and entomological indicators in Ebonyi State, Nigeria, 2017–2021

BackgroundIntense pyrethroid resistance threatens the effectiveness of the primary vector control intervention, insecticide-treated nets (ITNs), in Nigeria, the country with the largest malaria burden globally. In this study, the epidemiological and entomological impact of a new type of ITN (piperonyl-butoxide [PBO] ITNs) distributed in Ebonyi State were evaluated. The epidemiological impact was also compared to the impact of standard pyrethroid-only ITNs in Cross River State.MethodsA controlled interrupted time series analysis was conducted on monthly malaria incidence data collected at the health facility level, using a multilevel mixed-effects negative binomial model. Data were analysed two years before and after the PBO ITN campaign in Ebonyi State (December 2017 to November 2021). A pre-post analysis, with no comparison group, was used to assess the impact of PBO ITNs on human biting rates and indoor resting density in Ebonyi during the high transmission season immediately before and after the PBO ITN campaign.ResultsIn Ebonyi, PBO ITNs were associated with a 46.7% decrease (95%CI: -51.5, -40.8%; p < 0.001) in malaria case incidence in the 2 years after the PBO ITN distribution compared to a modelled scenario of no ITNs distributed, with a significant decrease from 269.6 predicted cases per 1000 population to 143.6. In Cross River, there was a significant 28.6% increase (95%CI: -10.4, 49.1%; p < 0.001) in malaria case incidence following the standard ITN distribution, with an increase from 71.2 predicted cases per 1000 population to 91.6. In Ebonyi, the human biting rate was 72% lower (IRR: 0.28; 95%CI 0.21, 0.39; p < 0.001) and indoor resting density was 73% lower (IRR: 0.27; 95%CI 0.21, 0.35; p < 0.001) after the PBO ITNs were distributed.ConclusionsThe epidemiological and entomological impact of the PBO ITNs underscore the impact of these ITNs in areas with confirmed pyrethroid resistance. These findings contribute to ongoing research on the impact of new types of ITNs in Nigeria, providing critical evidence for the Nigeria National Malaria Elimination Programme and other countries for future ITN procurement decisions as part of mass ITN campaign planning and malaria programming.

Investigation of resistance ratios and resistance mechanisms of Cydia pomonella (L.) (lepidoptera: Tortricidae) populations collected from apple orchards in Isparta (Türkiye) against some insecticides

The codling moth, Cydia pomonella is an important pest that causes significant economic losses in apples and walnuts in the world. The aim of this study was to investigate, for the first time in Türkiye, the resistance status and resistance mechanisms of codling moth, Cydia pomonella against indoxacarb, deltamethrin and emamectin benzoate. All apple orchard populations developed resistance ratios ranging from 3.38 to 22.37-fold to deltamethrin, 5.67–29.87-fold to indoxacarb and 1.46–3.05-fold to emamectin benzoate. The interaction of some synergists triphenyl phosphate (TPP), piperonyl butoxide (PBO) and diethyl maleate (DEM) with indoxacarb and deltamethrin was analyzed in the susceptible, MAREM and Tepeli populations with moderate resistance to indoxacarb and deltamethrin. While indoxacarb + TPP showed a significant synergistic effect only in MAREM population, a significant synergistic effect was observed with indoxacarb + TPP and indoxacarb + PBO in Tepeli population. The activities of detoxifying enzymes [esterase, glutathion –S– transferase (GST) and cytochrome P450 monooxygenase (P450)] studied by biochemical methods showed some variation depending on the population. The results of biochemical analyses showed that esterase and GST enzyme activities of all populations was between 0.51 and 0.94, 0.77 and1.29 mOD min−1mg−1 proteins, respectively. The P450 enzyme activities ranged from 0.53 to 0.78, RFU min−1mg−1 proteins. In addition, the L1014F knockdown mutation (CTT to TTT) corresponding to leucine to phenylalanine amino acid substitution of the voltage-gated sodium channel in Cydia pomonella was determined in MAREM and Tepeli populations. It was determined that while MAREM and Tepeli populations developed moderate resistance, the other populations developed a low level resistance to deltamethrin and indoxacarb. On the other hand, all populations developed a low level resistance to emamectin benzoate. The P450 and esterase enzyme activities were significantly higher in MAREM and Tepeli populations which were resistant to deltamethrin and indoxacarb than the susceptible population. In addition, a Kdr point mutation L1014F was detected in the deltamethrin resistant MAREM and Tepeli populations.

Efficacy of selected insecticide sprays against Indian meal moth (Lepidoptera: Pyralidae) wandering larvae on various surfaces

The Indian meal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), is a worldwide pest found in food product warehouses. Presence of P. interpunctella wandering larvae causes customer complaints and product rejection from customers. Its prepupal stage is susceptible to insecticide treatment due to their wandering behavior. We evaluated the efficacy of three insecticide sprays, ExciteR (0.017% pyrethrins, 0.174% piperonyl butoxide), Suspend SC (0.053% deltamethrin), and Transport Mikron (0.047% acetamiprid, 0.056% bifenthrin) against P. interpunctella wandering larvae by forced exposure to porous (vinyl tile, unpainted pinewood, concrete, and paper) and non-porous (steel, glazed ceramic) surfaces, and exposure to treated harborages made of two pieces of vinyl tiles, unpainted pinewood, and concrete. Under continuous exposure conditions, both ExciteR and Suspend SC caused significantly higher mortality (90–96%) than Transport Mikron (73–75%) on non-porous surfaces after 4 days (P < 0.05). But on porous surfaces, the three insecticides caused ≤63% mortality with Suspend SC being the most effective. When using 2× amount of insecticides as that in the previous test (2.1 gallon/1000 ft2, which is 4.33, 1.20, and 2.33 mg a.i./m2 of ExciteR, Suspend SC, and Transport Mikron, respectively), Suspend SC was the only insecticide causing high mortality (≥99%) through both forced exposure to concrete surface and exposure to treated concrete harborage. Whereas other treatments caused <12% mortality. Among the porous surfaces, residual efficacy of Suspend SC on concrete was significantly higher than that on vinyl tiles. Application of an effective liquid insecticide (such as 0.053% deltamethrin) into harborages or directly to the possible pupation sites, such as corners, cracks, and crevices could be an effective strategy for managing P. interpunctella wandering larvae in warehouse environment.

Detoxification Mechanism of Hinokitiol by Alternaria alternata and Its Application in Agricultural Antifungal Control.

Alternaria alternata is a common plant pathogen that can infect crops and reduce their production. In this work, an antagonism experiment between A. alternata and the essential oil of arborvitae (Platycladus orientalis) was performed, and it was proven that A. alternata had developed resistance to this plant-derived fungicide. A. alternata facilitated the biotransformation of hinokitiol (1), the main antifungal compound in the essential oil of arborvitae, into (R)-2-hydroxy-β-methylbenzeneethanol (2), which does not have antifungal activity against A. alternata. This biotransformation is an unusual ring-contraction reaction that was verified to be catalyzed by P450 enzyme hydroxylation and Baeyer-Villiger oxidation. In addition, the P450 enzyme inhibitors 1-aminobenzotriazole and piperonyl butoxide effectively prevented the destruction of the hinokitiol structure by A. alternata, and the combined use of these P450 enzyme inhibitors significantly increased the antifungal activity of hinokitiol. This work provides a theoretical reference for the further development of botanical fungicides.

Resistance monitoring of diamide insecticides and characterization of field-evolved chlorantraniliprole resistance among Chinese populations of the tomato pinworm Phthorimaea (=Tuta) absoluta (Lepidoptera: Gelechiidae)

The tomato pinworm, Phthorimaea (=Tuta) absoluta, is considered one of the most destructive and invasive insect pests worldwide, having developed significant resistance to many popular insecticides. In this study, we monitored the field resistance of P. absoluta populations from China to three diamide insecticides: flubendiamide, chlorantraniliprole, and cyantraniliprole. We found that one field population from Wuzhong City (WZ) exhibited high level of resistance to chlorantraniliprole. Using the WZ population and a susceptible reference strain (YN-S), we established a near-isogenic line (WZ-NIL) of P. absoluta with resistance to chlorantraniliprole. This strain also showed substantial cross-resistance to flubendiamide, and cyantraniliprole. Genetic analysis revealed that the inheritance of resistance to chlorantraniliprole in the WZ-NIL strain was autosomal and incompletely dominant. Additionally, the pesticide synergist piperonyl butoxide significantly inhibited chlorantraniliprole resistance by compromising P450 monooxygenase activity, which was significantly higher in the resistant strain. Furthermore, WZ-NIL had significantly prolonged developmental stages, lower pupation rates, reduced female fecundity, and lower egg hatchability than YN-S individuals. The fitness of WZ-NIL relative to YN-S was estimated to be 0.73, indicating significant fitness cost associated with chlorantraniliprole resistance. Rotating chlorantraniliprole with other insecticides that have different modes of action and degradation may be particularly useful for managing chlorantraniliprole resistance in P. absoluta.

Roe deer exposure to trace metals and pesticides in forests and agricultural plains of North-eastern France.

Human activities related to agriculture and industrial development result in the emission of different types of pollutants in the environment. The phytosanitary pressure depends on the time of the year, the type of habitat, and the used treatments. Wildlife, particularly ungulates, can be exposed to pesticides and trace metals through their herbivorous diet directly impregnated by the environmental contamination. Wild game thus plays a sentinel role but can also represent a risk for human health by exposing venison consumers to these contaminants. The present study explored this dual problem in two types of habitat: forests and agricultural plains located in North-eastern France. Samples of liver and muscle were taken from ten roe deer, five caught on the forests and five others in the plains, to determine contaminant levels. There was no significant difference in contamination of roe deer livers according to the habitat for three trace metals (chromium, nickel, and copper), while lead concentrations were higher in samples from the plain (0.85 vs. 0.74mg·kg-1 dry matter DM; P < 0.01). Conversely, zinc concentrations were higher in forest roe deer (110 vs. 95mg·kg-1 DM; P < 0.05). Cadmium concentrations did not differ significantly between the two habitats, but forest samples showed strong inter-individual variability, with some values close to those observed on the plains and others higher (1.8 vs. 0.3mg·kg-1 DM for forest and plain, respectively; P = 0.11). Mercury was not detected in any sample. The majority of pesticides investigated were not detected. Only a few pesticides were in trace amount (from 1 to 8µg·kg-1 fresh matter), either in the liver (e.g., heptachlor and oxychlordane) or in both tissues, liver and muscle (e.g., pyridalyl). Some compounds were only detected in plain (e.g., cypermethrin and fenpropidin) and others in forest (piperonyl butoxide and pyridalyl) samples. Analyses mainly detected insecticides in samples taken from forests and agricultural plains. Fungicides and a few herbicides were mainly detected in plain sampled animals. Pesticides and copper concentrations in roe deer tissues were lower than maximum residue levels established for farm (copper) or wild terrestrial (pesticides) animals. Cadmium concentrations were also lower than the maximum level for most samples, except for some forest roe deer. However, both forest and plain roe deer exceeded the maximum level for lead in liver. Human exposure was simulated according to different amounts of meat consumed and compared with toxicological reference values. This study highlighted an overall low risk to human health for both pesticides, copper and lead, with more reservations about cadmium for some roe deer shot in forest. The differences in roe deer contamination between forests and agricultural plains for some trace elements and organic pollutants confirmed the sentinel role of this species.

Power calculation for mosquito bioassays: Quantifying variability in the WHO tube bioassay and developing sample size guidance for the PBO synergism assay using a Shiny application

Background The WHO tube bioassay is a method for exposing mosquitos to determine susceptibility to insecticides, with mortality to discriminating doses &lt;98% indicating possible resistance and &lt;90% confirming resistance. This bioassay is also used for synergism testing to assess if susceptibility is restored by pre-exposure to the synergist piperonyl butoxide. Methods Here we perform testing with pyrethroid-susceptible and pyrethroid-resistant An. gambiae to quantify the variability of the WHO tube bioassay and identify its sources. These estimates of within and between day variability are then used to evaluate the power of the bioassay to detect a mortality difference between pyrethroid-only and pyrethroid-PBO. Results We show that approximately two-thirds of variation occurs between days, with the pyrethroid-susceptible strain twice as variable as the pyrethroid-resistant strain. The total number of mosquitoes in the tube and their bodyweight contributes to approximately 10% of this variability. Changes in temperature and humidity, within a climate-controlled insectary, didn’t impact mortality. Using a simulation-based framework, we show that the current synergism guidelines, using a 4x4 design, can reliably detect a difference between 90% and 100% mortality (&gt;90% power). However, as the mortality of either group gets closer to 50%, a 10% difference between groups is more difficult to reliably detect. In the worst-case scenario where the mortality of either group is 50%, the mortality difference must be &gt;22.5% to be detected with 80% power. We provide an R shiny application to assess power for other comparisons. Conclusions Our findings indicate that detecting synergism with the WHO tube assay is more difficult than assumed by the current WHO guidelines. Additionally, we demonstrate the value of using a Shiny application to make the outputs of simulation-based power analysis readily available to end-users, allowing them to determine the number of tubes needed to detect a given mortality difference.

Bioaccumulation of Deltamethrin and Piperonyl butoxide in Labeo rohita fish

Deltamethrin (DLM), in combination with the synergist piperonyl butoxide (PBO), is extensively used in pest control programs due to its potent pesticidal properties and appreciable safety margin. However, various research studies report their adverse effects on non-target organisms. In this study, we investigated the toxicity of DLM, PBO, and a DLM-PBO (3:1) combination on Labeo rohita (L. rohita) fish fingerlings. Fish behavior and mortality rates were recorded at different time intervals up to 96 h for concentrations of 0.003, 0.007, 0.015, 0.031, and 0.062 µg/mL, respectively. Biochemical, hematological, and histopathological studies were carried out. High-performance liquid chromatography (HPLC) was used to detect and quantify residues in fish samples. The LC50 values after 48 h for DLM, PBO, and DLM-PBO exposed fish fingerlings were found to be 0.028, 0.066, and 0.007 µg/mL, respectively. At a concentration of 0.003 µg/mL of DLM, PBO, and DLM-PBO, the treated fish fingerlings exhibited similar behavior to the control group. Hematological parameters, such as red blood cell (RBC) and white blood cell (WBC) counts, were reduced in the treated groups compared to the control. Biochemical parameters showed increased levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), while total serum protein levels decreased in DLM, PBO, and DLM-PBO treated fingerlings. Histopathological examination of liver, gill, and heart tissues revealed lesions with hydropic degeneration in the liver and fusions of gill lamellae in the treated tissues. Fish fingerlings exposed to the DLM-PBO combination appeared highly prone to toxicity compared to those treated with DLM and PBO separately.

Examination of piperonyl butoxide developmental toxicity as a Sonic hedgehog pathway inhibitor targeting limb and palate morphogenesis

Piperonyl butoxide (PBO) is a pesticide synergist with widespread use and human exposure that was discovered to inhibit Sonic hedgehog (Shh) signaling, a pathway required for numerous developmental processes. Previous examinations of PBO’s potential for developmental toxicity have generated seemingly conflicting results. We investigated the impact of acute PBO exposure targeting Shh pathway activity during palate and limb morphogenesis. Timed-pregnant C57BL/6 J mice were exposed to a single PBO dose (67–1800 mg/kg) at gestational day (GD) 9.75, and litters were collected at GD10.25 and GD10.75 to examine Shh pathway activity or GD17 for phenotypic assessment. PBO exposure induced dose-dependent limb malformations and cleft palate in the highest dose group. Following PBO exposure, reduced expression of the Shh pathway activity markers Gli1 and Ptch1 was observed in the embryonic limb buds and craniofacial processes. These findings provide additional evidence that prenatal PBO exposure targeting Shh pathway activity can result in malformations in mice that parallel common etiologically complex human birth defects.

Piperonyl butoxide elicits a robust transcriptional response in adult Drosophila melanogaster

While much attention has been devoted to understanding the transcriptional changes underlying resistance to insecticides, comparatively little is known about the transcriptional response of naive insects to agrochemicals. In this study, we analyze the transcriptomic response of an insecticide susceptible strain of Drosophila melanogaster to nine agrochemicals using a robust method that goes beyond classical replication standards. Our findings demonstrate that exposure to piperonyl butoxide (PBO), but not to eight other compounds, elicits a robust transcriptional response in a wild-type strain of Drosophila melanogaster. PBO exposure leads to the upregulation of a subset of Cyps, GSTs, UGTs and EcKls. This response is both time and concentration-dependent, suggesting that the degree of inhibition of P450 activity correlates with the magnitude of the transcriptional response. Furthermore, the upregulation of these enzymes is excluded from reproductive organs. Additionally, different sets of genes are regulated in the digestive/secretory tract and the carcass. Our results suggest that P450s play a role in metabolizing yet unidentified endogenous compounds and are involved in an as-yet-unknown physiological regulatory feedback loop.

Biochemical and molecular characterization of pyrifluquinazon resistance in Bemisia tabaci Asia I

One of the most devastating polyphagous pests in the world, Bemisia tabaci (Gennadius), causes significant damage to a variety of crops and has demonstrated the ability to develop resistance to different classes of insecticides rapidly. A novel pyridine azomethine derivative pyrifluquinazon exhibits exceptional insecticidal toxicity against B. tabaci by interrupting the function of chordotonal receptor neurons. Formulating resistance management strategies is crucial to ensure the long-term use of this insecticide for whitefly control; however, the characteristics and possible mechanisms of pyrifluquinazon resistance in B. tabaci remain unclear. By employing pyrifluquinazon selection for 22 successive generations, the pyrifluquinazon-resistant strain (PQZ-R) was generated from a laboratory-susceptible population of B. tabaci Asia I (Lab-WB) and exhibited 39.65-fold resistance to pyrifluquinazon. When the realized heritability (h2) of B. tabaci to pyrifluquinazon was assumed to be the laboratory-estimated value (h2 = 0.181) and the mortality was 70–90%, only 12.3–22.6 generations were expected to be required to obtain a 10-fold increase in pyrifluquinazon resistance. While there was no significant cross-resistance to cyantraniliprole, dinotefuran, flonicamid, flupyradifurone, pymetrozine, sulfoxaflor, or thiamethoxam, the PQZ-R strain displayed slight cross-resistance to afidopyropen (3.14-fold). Synergism tests indicated that piperonyl butoxide (PBO) inhibits (4.36-fold) pyrifluquinazon resistance in the PQZ-R strain. This impact may be attributed to enhanced detoxification (elevated 3.91-fold) mediated by cytochrome P450 monooxygenase (P450). Compared to Lab-WB, the PQZ-R strain exhibited no significant overexpression of the 13 published detoxification-related P450 genes from CYP303, CYP4, and CYP6 families in B. tabaci. The combined knowledge gained from this study will enable further investigations into the function of qualitative and quantitative variations in P450-encoded genes, leading to innovative approaches for efficiently managing B. tabaci.

High concentrations of piperonyl butoxide (PBO) enhance toxicity of S-methoprene against the lesser grain borer, Rhyzopertha dominica.

Insect growth regulators (IGRs) have been playing a major role in the effective management of a range of stored product insect pests including species that have developed resistance to major groups of insecticides, such as organophosphates (OPs) and synthetic pyrethroids (SPs). In the present study, we evaluated the efficacy of S-methoprene alone and in combination with piperonyl butoxide (PBO), an adjuvant component of insecticides for synergy, against two strains, Lab-S (susceptible) and Met-R (Methoprene resistant) of an economically important pest species, the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae). Adults of both Lab-S and Met-R strains were exposed to wheat treated with multiple concentrations of S-methoprene ranging from 0.001 to 0.01 and 10 to 60 mg/kg, respectively, alone and in combination with PBO. A variety of concentrations, including 0.27, 0.53, 0.80, and 1.07 g/kg, were evaluated for PBO. Mortality of adults and percent reduction in progeny were assessed after 14 and 65 days of treatment, respectively. As anticipated, the adult mortality rates of both strains were not significantly affected by S-methoprene alone. However, the number of progeny produced confirmed that the Met-R strain exhibited a high level of resistance to S-methoprene alone, with over 50 F1 progeny adults surviving in wheat treated with the maximal rate, 10 mg/kg. In contrast, the toxicity of S-methoprene was increased against the same resistant strain (Met-R), by 0.80 or 1.07 g/kg of PBO in combination treatment, resulting in a significant reduction in progeny numbers (25 adults per container). Although the tested concentrations of S-methoprene and PBO were well above the currently registered rate globally, our results highlight the fact that PBO enhances the toxicity of S-methoprene to some extent, reaffirming that the mode of action of the latter involves the inhibition of mixed-function oxidases (MFOs) and highlights the need for further research into developing potential binary or triplet formulations containing these two active ingredients (AIs).

A novel mutation Trp-2027-Gly in acetyl-CoA carboxylase confers resistance to cyhalofop-butyl in Chinese sprangletop (Leptochloa chinensis).

Chinese sprangletop [Leptochloa chinensis (L.) Nees] control is threatened by resistance to acetyl-CoA carboxylase (ACCase)-inhibiting herbicides. In this study, a L. chinensis population, HFLJ18, that survived cyhalofop-butyl [aryloxyphenoxypropionate (APP) herbicide, CyB] treatment was collected from a rice field in Lujiang County, Anhui Province, China. This study aimed to evaluate the susceptibility of HFLJ18 to herbicides with different modes-of-action and investigate the potential mechanisms of resistance to CyB. The HFLJ18 population exhibited high levels of resistance to CyB (10.92-fold) and showed resistance to the ACCase inhibitors metamifop (4.63-fold) and fenoxaprop-P-ethyl (8.39-fold), but was susceptible to clethodim, pinoxaden, florpyrauxifen-benzyl, oxadiazon and pretilachlor. Target gene sequencing revealed a novel Trp-to-Gly substitution at codon position 2027 of ACCase in the resistant plants. Molecular docking revealed that the spatial structure of ACCase changed significantly following the substitution, as indicated by reduced H-bonds. A newly derived cleaved amplified polymorphic sequence (dCAPS) marker was subsequently developed to detect the Trp-2027-Gly mutation in the ACCase of L. chinensis. Additionally, pretreatment with the cytochrome P450 (P450) inhibitor piperonyl butoxide (PBO) and the glutathione S-transferase (GST) inhibitor 4-chloro-7-nitrobenzoxadiazole (NBD-Cl) did not reverse resistance to CyB, suggesting that nontarget-site resistance mechanisms were not involved in CyB resistance in the HFLJ18 population. Overall, the resistance to CyB in the HFLJ18 population derived from the mutation of ACCase gene, and to the best of our knowledge, this is the first report of the ACCase Trp-2027-Gly mutation conferring resistance to ACCase-inhibiting herbicides in grass species. © 2024 Society of Chemical Industry.

Evaluation of Adulticidal Efficacy of Cypermethrin, Tetramethrin, and Piperonyl Butoxide in Housefly (Musca Domestica)

Introduction: Houseflies serve as vectors of diseases in animals and humans. Chemical pesticides are a practical way of eliminating insects; however, resistance to insecticides has been reported. This study aims to evaluate the adulticidal efficacy of cypermethrin, tetramethrin, and piperonyl butoxide in housefly. Methods: Adult flies were cultured within net enclosures using completely randomized design with four replications each treatment. Treatment A was water and served as the negative control, treatment B was cypermethrin + tetramethrin + piperonyl butoxide 32% EC and served as the positive control, and treatment C was cypermethrin + tetramethrin + piperonyl butoxide 48% EC. The number of dead houseflies were counted 15, 30, 60, and 120 minutes post- application of treatments. The results were analyzed through analysis of variance (P≤0.05) followed by Tukey’s HSD (honestly significant difference). Results and Discussion: The values for the cumulative mean and percentage of killed houseflies were all significantly different from each other, with treatment C showing the highest values, followed by treatment B, then treatment A. The maximum adulticidal effect of treatment C was apparent within 15 minutes post-application, while the maximum adulticidal effect of treatment B was observed within 30 minutes after application. Conclusion: Results suggest that treatment C was a more effective adulticidal than other groups. Using cypermethrin + tetramethrin + piperonyl butoxide 48% EC can be the solution to the problem, but it should be noted that future resistance may occur. Increased concentrations may also imply environmental, human, and animal health hazards.

The sublethal impacts of five insecticidal formulations on Oryzaephilus surinamensis behavioral traits.

Mortality caused by various pyrethroids, and neonicotinoids has been studied for stored-product insects in the past, yet limited information exists on the sublethal effects they can induce to Oryzaephilus surinamensis. In the current study, the sublethal effects of deltamethrin, λ-cyhalothrin, α-cypermethrin, etofenprox, and the mixture of acetamiprid with d-tetramethrin and piperonyl butoxide on the mobility of O. surinamensis in the presence or the absence of a food source was investigated. Lethal concentrations (LCs) were lower for deltamethrin, α-cypermethrin, and λ-cyhalothrin (LC10 = 0.000233, 0.000211, and 0.000271 mg active ingredient (a.i.) cm-2, LC30 = 0.000413, 0.000398, and 0.000447 mg a.i. cm-2, respectively), followed by etofenprox, and the mixture of acetamiprid with d-tetramethrin and piperonyl butoxide (LC10 = 0.00228 and 0.003267 mg a.i. cm-2, LC30 = 0.00437 and 0.01188 mg a.i. cm-2, respectively). Deltamethrin and λ-cyhalothrin negatively impacted adult walking behavior, increasing stop durations compared to controls. Adults exposed to LC10 and LC30 of λ-cyhalothrin, and LC30 of deltamethrin exhibited prolonged periods on their backs compared to the remaining treatments and the controls. The α-cypermethrin LC30-exposed adults exhibited significantly shorter walking and stopping durations than controls but demonstrated prolonged climbing on the arena walls compared to adults exposed to the remaining a.i. and the control. A similar trend was observed for etofenprox. Under sublethal concentrations, λ-cyhalothrin and deltamethrin increased stop intervals and reduced the duration of climbing attempts of O. surinamensis versus α-cypermethrin. These findings advance comprehension of the underexplored sublethal impacts of the tested a.i. on O. surinamensis adults, holding potential for leveraging insecticide-induced behavioral effects to enhance warehouse pest management. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Biochemical and genetic mechanisms in Pieris rapae (Lepidoptera: Pieridae) resistance under emamectin benzoate stress

Pieris rapae (Lepidoptera: Pieridae) poses a significant threat to Brassicaceae crops, leading to substantial losses annually. Repeated insecticide applications are widely used to protect crops and increase the resistance of P. rapae. Exploring the biochemical and molecular basis of insecticide tolerance in P. rapae is crucial for achieving effective insect suppuration and implementing resistance control strategies. In our research, emamectin benzoate (EBZ) resistance was developed in P. rapae strain through selective pressure over 15 generations. Moreover, the biochemical mechanisms underlying resistance to EBZ and its potential cross-resistance to other insecticides were studied. Additionally, the expression levels of cytochrome P450 (CYP450) and glutathione-s-transferase (GST) genes in P. rapae were quantitatively assessed upon exposure to EBZ using real-time PCR. Our data exhibited that the LC50 value of susceptible strain (Sus) and EBZ resistance strain (EBZ-R) were 0.009 and 8.09 mg/L, with a resistance ratio (RR) reaching 898.8-fold. The EBZ-R stain displayed notably low cross-resistance to lambda-cyhalothrin, spinetoram, and cypermethrin. However, it demonstrated a moderate level of cross-resistance to deltamethrin. Conversely, no cross-resistance was noted to chlorantraniliprole and indoxacarb. Notably, enzyme inhibitors of detoxification enzymes revealed that piperonyl butoxide (PBO) and diethyl maleate (DEM) enhanced the EBZ toxicity to the resistant strain, indicating the potential involvement of CYP450 and GST in avermectin resistance. A remarkable enhancement in CYP450 and GST activity was observed in the EBZ-R stain. CYP450 and GST genes are upregulated in the EBZ-R stain compared to the Sus strain, which serves as a basis for comprehending the mechanism behind P. rapae resistance to EBZ. The molecular docking analysis demonstrated that EBZ has a high binding affinity with CYP6AE120 and PrGSTS1 with docking energy values of −20.19 and −22.57 kcal/mol, respectively. Our findings offer valuable insights into crafting efficient strategies to monitor and manage resistance in P. rapae populations in Egypt.

Metabolism of a fungicide propiconazole by Cunninghamella elegans ATCC36112.

The metabolic breakdown of propiconazole by fungi was examined, and it was found that the microbial model (Cunninghamella elegans ATCC36112) efficiently degrades the triazole fungicide propiconazole through the action of cytochrome P450. This enzyme primarily facilitates the oxidation and hydrolysis processes involved in phase I metabolism. We observed major metabolites indicating hydroxylation/oxidation of propyl groups of propiconazole. Around 98% of propiconazole underwent degradation within a span of 3days post-treatment, leading to the accumulation of five metabolites (M1-M5). The experiments started with a preliminary identification of propiconazole and its metabolites using GC-MS. The identified metabolites were then separated and identified by in-depth analysis using preparative UHPLC and MS/MS. The metabolites of propiconazole are M1 (CGA-118245), M2(CGA-118244), M3(CGA-136735), M4(GB-XLIII-42-1), and M5(SYN-542636). To further investigate the role of key enzymes in potential fungi, we treated the culture medium with piperonyl butoxide (PB) and methimazole (MZ), and then examined the kinetic responses of propiconazole and its metabolites. The results indicated a significant reduction in the metabolism rate of propiconazole in the medium treated with PB, while methimazole showed weaker inhibitory effects on the metabolism of propiconazole in the fungus C. elegans.

Efficacy of Interceptor G2, Royal Guard and PermaNet 3.0 against pyrethroid-resistant Anopheles gambiae s.l. from Za-Kpota, southern Benin: an experimental hut trial

BackgroundThe widespread use of insecticide-treated nets (ITNs) has significantly contributed to the reduction in malaria cases and deaths observed across Africa. Unfortunately, this control strategy is threatened by the rapid spread of pyrethroid resistance in malaria vectors. Dual-active-ingredient insecticidal nets are now available to mitigate the impact of pyrethroid resistance. To facilitate evidence-based decisions regarding product selection in specific use settings, data are needed on the efficacy of these different nets against local mosquito populations.MethodsTwo experimental hut trials were performed in Za-Kpota, southern Benin in 2021 to evaluate the performance of Interceptor G2 (BASF), Royal Guard (Disease Control Technologies) and PermaNet 3.0 (Vestergaard Frandsen), all dual-active-ingredient bednets, in comparison to untreated or standard pyrethroid-treated bednets, against free-flying wild Anopheles gambiae mosquitoes. The performance of some of these next-generation nets was compared to the same type of nets that have been in use for up to 2 years. Mosquitoes collected in the huts were followed up after exposure to assess the sublethal effects of treatments on certain life-history traits.ResultsThe predominant species in the study site was Anopheles gambiae sensu stricto (An. gambiae s.s.). Both Anopheles coluzzii and An. gambiae s.s. were resistant to pyrethroids (deltamethrin susceptibility was restored by piperonyl butoxide pre-exposure). In the experimental hut trials, the highest blood-feeding inhibition (5.56%) was recorded for the Royal Guard net, relative to the standard PermaNet 2.0 net (44.44% inhibition). The highest 72-h mortality rate (90.11%) was recorded for the Interceptor G2 net compared to the PermaNet 2.0 net (56.04%). After exposure, the risk of death of An. gambiae sensu lato (An. gambiae s.l.) was 6.5-fold higher with the Interceptor G2 net and 4.4-fold higher with the PermaNet 3.0 net compared to the respective untreated net. Lower mosquito mortality was recorded with an aged Interceptor G2 net compared to a new Interceptor G2 net. Oviposition rates were lower in mosquitoes collected from huts containing ITNs compared to those of untreated controls. None of the mosquitoes collected from huts equipped with Royal Guard nets laid any eggs.ConclusionsThe Royal Guard and Interceptor G2 nets showed a potential to significantly improve the control of malaria-transmitting vectors. However, the PermaNet 3.0 net remains effective in pyrethroid-resistant areas.Graphical