Long-term trends in housefly (Musca domestica L.) insecticide resistance in China.

Controlling housefly populations relies on the use of insecticides, which inevitably leads to the development of resistance. A better and more comprehensive understanding of the spatial and temporal distribution of resistance could guide the control of houseflies. However, most studies on housefly resistance in China are scattered and poorly coordinated. We collected resistance data from houseflies in the published literature and from the vector biomonitoring system of the Chinese Center for Disease Control and Prevention. A 5- or 10-year resolution was used to study the temporal dynamics of resistance to five commonly used insecticides: deltamethrin, permethrin, beta-cypermethrin, dichlorvos, and propoxur. ArcGIS was used to visualize their spatial distributions. The correlation between year and resistance coefficient was determined using SPSS 26.0 and RStudio to explore the changes in resistance over the years. A total of 2128 data were included in this study, ranging from 1982 to 2022, based on which we found significant increases in resistance over the past forty years for the five studied insecticides. Among them, pyrethroids had the most strikingly elevated resistance level and were mainly distributed in the northern and southeastern coastal areas. Dichlorvos and propoxur had intermediate increases in resistance, and most of these increases were identified in North China and the Yangtze River. Housefly resistance to commonly used insecticides in China is increasing and spatially heterogeneous. This finding also highlights the necessity of continuous routine surveillance of housefly resistance, which could guide future housefly control operations and slow the development of resistance.

Monitoring Aedes populations for arboviruses, Wolbachia, insecticide resistance and its mechanisms in various agroecosystems in Benin.

Aedes mosquitoes are the main vectors of arboviruses in Benin. Cases of dengue have been reported in Benin with all four serotypes of the virus actively circulating in this region. Some agricultural settings are known to harbor Aedes vectors responsible for the transmission of arboviruses. The massive use of certain insecticides in agricultural settings has probably contributed to insecticide resistance in these vectors. In Benin, the susceptibility of arbovirus vectors to insecticides is poorly studied. In addition, the distribution of Wolbachia spp., which is used against some arboviruses is unknown. Moreover, there is limited information regarding the vectors responsible for the transmission of arboviruses in Benin. This present study monitored the species composition, arboviruses, and Wolbachia symbiont status, as well as the phenotypic and molecular insecticide resistance profile of Aedes populations from three agroecosystems in Benin. Aedes species identification was performed morphologically and confirmed using qPCR. (RT)-qPCR assay was applied for monitoring the presence of DENV, CHIKV, ZIKV, and WNV pathogens as well as for naturally occurring Wolbachia symbionts. Insecticide resistance was assessed phenotypically, by permethrin (0.75%) exposure of Adults (F0) using World Health Organization (WHO) bioassay protocols, and at the molecular level, using TaqMan (RT)-qPCR assays for assessing knock-down resistance (kdr) mutations (F1534C, V1016G/I, and S989P) and the expression levels of eight detoxification genes (P450s from the CYP9 and CYP6 families, carboxylesterases and glutathione-S-transferases). Aedes aegypti (Ae. aegypti) mosquitoes were the most abundant (93.9%) in the three agroecosystems studied, followed by Aedes albopictus (Ae. albopictus) mosquitoes (6.1%). No arboviruses were detected in the study's mosquito populations. Naturally occurring Wolbachia symbionts were present in 7 pools out of 15 pools tested. This could influence the effectiveness of vector control strategies based on exogenously introduced Wolbachia, all present in the three agroecosystems. Full susceptibility to permethrin was observed in all tested populations of Ae. albopictus. On the contrary, Ae. aegypti were found to be resistant in all three agroecosystem sites except for banana plantation sites, where full susceptibility was observed. Molecular analysis revealed that individual target site resistance kdr mutations F1534C and V1016G/I were detected in most Ae. aegypti populations. Additionally, double mutant (F1534C + V1016G/I) mosquitoes were found in some populations, and in one case, triple mutant (F1534C + V1016G/I + S989P) mosquitoes were detected. Metabolic resistance, as reflected by overexpression of three P450 genes (CYP6BB2, CYP9J26, and CYP9J32), was also detected in Ae. aegypti mosquitoes. Our study provides information that could be used to strategize future vector control strategies and highlights the importance of continuing vector surveillance. Future studies should assess the effect of piperonyl butoxide (PBO) on metabolic resistance and identify the different strains of Wolbachia spp., to choose the best vector control strategies in Benin.

Insecticide resistance status of Aedes aegypti in border areas of Yunnan Province.

BACKGROUND: Aedes aegypti is a main vector of arboviral diseases, principally dengue, chikungunya, and Zika. Insecticides remain the most effective vector control method. Pyrethroid is the main insecticide currently used, and the long-term use of insecticides can cause mosquitoes to develop knockdown resistance. Studying the mutation sites and genotypes of Ae. aegypti can reveal the mutation characteristics and regional distribution of the kdr gene in an Ae. aegypti population. Testing for a correlation between the mutation rate in various populations and pyrethrin resistance can clarify the resistance mechanism. RESULTS: The bioassay results showed that all 15 populations are resistant. In the study of the kdr gene, three non-synonymous mutations were identified in the DNA of first generation females from the wild Ae. aegypti population: S989P (TCC-CCC), V1016G (GTA-GGA), and F1534C (TTC-TGC). The mortality rate of the various populations was correlated with the mutation rate at the V1016G + F1534C locus, but not the S989P + V1016G locus. CONCLUSION: Aedes aegypti populations in border regions of Yunnan Province are resistant to permethrin and beta-cyfluthrin. The insecticidal effect of beta-cyfluthrin is stronger than that of permethrin. The mutation rate at sites V1016G + F1534C is negatively correlated with the mortality of Ae. aegypti based on bioassays. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Lack of fitness costs associated with resistance to permethrin in Musca domestica.

Resistance to permethrin has been reported in Pakistani strains of Musca domestica. The present study explored the performance of biological traits and analyzed life tables to determine whether there is any detrimental effect of permethrin resistance on the fitness of permethrin-resistant strains [an isogenic resistant strain (Perm-R) and a field strain (Perm-F)] compared to a susceptible strain (Perm-S). Perm-R and Perm-F exhibited 233.93- and 6.87-fold resistance to permethrin, respectively. Life table analyses revealed that the Perm-R strain had a significantly shorter preadult duration, longer longevity, shorter preoviposition period, higher fecundity, finite rate of increase, intrinsic rate of increase, net reproductive rate and a shorter mean generation time, followed by the Perm-F strain when compared to the Perm-S strain. Data of the performance of biological traits reveled that permethrin resistance strains had a better fit than that of the Perm-S strain. The enhanced fitness of resistant strains of M. domestica may accelerate resistance development to permethrin and other pyrethroids in Pakistan. Some possible measures to manage M. domestica and permethrin resistance in situations of fitness advantage are discussed.

Differences in excito-repellent behavior between susceptible and permethrin-resistant strains of Aedes aegypti upon exposure to insecticide-treated fabric.

BACKGROUND: An important component of the biological activity of pyrethroids, when used in disease vector control, is excito-repellency. In this study, behavioral differences between insecticide susceptible (Orlando) and pyrethroid resistant (Puerto Rican) strains of Aedes aegypti were explored in a round glass arena using fabrics treated with permethrin, etofenprox, deltamethrin, or DDT. Repellency was evaluated across several variables, including the time to first flight (TFF), number of landings (NOL), total flight time (TFT), and maximum surface contact (MSC), all by video analysis. RESULTS: Results from the Orlando strain indicated they were less likely than the Puerto Rican strain to tolerate tarsal contact with the treated fabrics. All four response variables indicated that the mosquito flight and landing behavior was most affected by pyrethroid resistance [knockdown resistance (kdr)] status. In other experiments, mosquitoes were surgically altered, with antennae ablated bilaterally, and these mosquitoes were more likely to stay on the treated surfaces for longer periods of time, irrespective of any chemical exposure. There were also differences in the responses to antennal ablation between the two strains of mosquitoes, indicating that resistance factors, probably kdr, influence the reactivity of mosquitoes to pyrethroid and DDT treatments, and that it was not completely negated by antennal ablation. CONCLUSIONS: These findings confirm the role of antennal olfactory components in the expression of excito-repellent behaviors, and also support the hypothesis that excito-repellency from pyrethroid/DDT exposure is probably due to a combination of sublethal neurotoxic excitation and interactions with the olfactory system. © 2023 Society of Chemical Industry.

Prevalence of Permethrin Resistance in Culex Tarsalis Populations in Southern California.

In the western United States, Culex tarsalis is the most important vector of West Nile virus. Insecticides containing permethrin or other pyrethroid compounds are commonly used to control these mosquitoes. Because of the range of environments where Cx. tarsalis are found, this species is under insecticide pressure from both vector control and agricultural spraying. Mosquito populations may evolve resistance through mechanisms such as target site insensitivity, including the frequently identified knockdown resistance (kdr) mutations. Prevalence of permethrin resistance was determined for Cx. tarsalis from 5 southern California field sites representing 2 distinct valley regions (Coachella Valley and Inland Valley), which are geographically separated by the north-south-running Peninsular Mountain Ranges. These two valley regions are >100 km apart and vary considerably in their environmental and habitat characteristics. Permethrin resistance in mosquito populations was determined by the Centers for Disease Control and Prevention (CDC) bottle bioassay, using glass bottles coated with permethrin at 0.19 µg/cm2 of internal surface. Permethrin resistance was evident in Cx. tarsalis populations from the Coachella Valley field sites with all sites showing similar mortality in the bottle bioassay, while Cx. tarsalis from the Inland Valley field sites were largely susceptible to permethrin, with mortality rates that were similar to a susceptible lab strain of Cx. tarsalis.

Resistance and inhibitor testing on Aedes aegypti (Linnaeus) (Culicidae: Diptera) populations in the Florida Keys.

Aedes aegypti is the species of greatest concern for mosquito-borne disease in the Florida Keys. Previous locally transmitted dengue outbreaks in Key West (2009-2010) and Key Largo (2020) illustrate the need for an immediate and effective response plan to maintain Ae. aegypti populations below threshold levels. An important part of the Florida Keys Mosquito Control District's vector response plan is adulticide application because it can provide an immediate reduction in Ae. aegypti adults in the community. It has become apparent that in the Florida Keys, and throughout Florida, Ae. aegypti resistance to the adulticide permethrin is prevalent. This study uses the CDC bottle bioassay method to look at resistance in Ae. aegypti collected from Key Largo, Vaca Key, and Key West, FL. Resistance was found in all three populations when exposed to permethrin and Sumithrin® but not malathion. Inhibitor testing revealed that esterase and glutathione transferase activity is involved in resistance to permethrin in Key Largo and Key West Ae. aegypti populations while oxidase activity is involved in resistance to permethrin in Ae. aegypti from Vaca Key. Lack of knockdown at the diagnostic time and previous studies detecting the presence of kdr-associated allele mutations suggest knockdown resistance in all three populations. Results from this study show that there are multiple factors involved with resistance in the Ae. aegypti populations in the Florida Keys and that resistance mechanisms vary between islands. Continued surveillance will remain important so the most effective active ingredients can be used in response to future disease transmission.

Vegetable oil-based surfactants are adjuvants that enhance the efficacy of neonicotinoid insecticides and can bias susceptibility testing in adult mosquitoes.

BACKGROUND: The standard operating procedure for testing the susceptibility of adult mosquitoes to neonicotinoid or butenolide insecticides recommends using a vegetable oil ester (Mero) as a surfactant. However, there is growing evidence that this adjuvant contains surfactants that can enhance insecticide activity, mask resistance and bias the bioassay. METHODOLOGY/PRINCIPAL FINDINGS: Using standard bioassays, we tested the effects of commercial formulations of vegetable oil-based surfactants similar to Mero on the activity of a spectrum of active ingredients including four neonicotinoids (acetamiprid, clothianidin, imidacloprid and thiamethoxam) and two pyrethroids (permethrin and deltamethrin). We found that three different brands of linseed oil soap used as cleaning products drastically enhanced neonicotinoid activity in Anopheles mosquitoes. At 1% (v/v), the surfactant reduced the median lethal concentration, LC50, of clothianidin more than 10-fold both in susceptible and in resistant populations of Anopheles gambiae. At 1% or 0.5% (v/v), linseed oil soap restored the susceptibility of adult mosquitoes fully to clothianidin, thiamethoxam and imidacloprid and partially to acetamiprid. By contrast, adding soap to the active ingredient did not significantly affect the level of resistance to permethrin or deltamethrin suggesting that vegetable oil-based surfactants specifically enhance the potency of some classes of insecticides. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that surfactants are not inert ingredients, and their use in susceptibility testing may jeopardize the ability to detect resistance. Further research is needed to evaluate the potential, the limitations and the challenges of using some surfactants as adjuvants to enhance the potency of some chemicals applied in mosquito control.

Pyrethroid insecticides susceptibility profiles and evaluation of L1014F kdr mutant alleles in Culex quinquefasciatus from lymphatic filariasis endemic communities.

This study investigated the dynamics in pyrethriod resistance and the presence/frequencies of L1014F knockdown resistance mutant allelles in Culex quinquefasciatus vector populations from Uruan Local Government Area of AkwaIbom State, Southern Nigeria between the months of March and November, 2021. Uruan LGA is among the endemic LGAs for lymphatic filariasis in AkwaIbomState. Female Anopheles mosquitoes from Eman Uruan, Ituk Mbang and Idu Uruan were exposed to permethrin, deltamethrin and alphacypermethrin in CDC insecticide coated bottles for susceptibility bioassay following standard protocols. The mosquitoes were obtained as aquatic forms from the study sites and reared under laboratory conditions to adults. The adult mosquitoes were used for this study. All the mosquitoes used for the insecticide susceptibility bioassay were morphologically identified. Standard Polymerase chain reaction (PCR) was used for authenticating the Culex quinquefasciatus species. A portion of the vgsc (917 bp) gene spanning the entire intron and the exon containing the L1014F mutation associated with knockdown resistance (kdr) in the vectorswere amplified using Allele-SPECIFIC POLYMERASE CHAIN REACTION (AS-PCR) in order to detect target site insensitivity in the vectors from the study sites. Results obtained revealed that vectors from all the study sites were resistant to permethrin insecticide (mortality rate: 18-23%). Suspected resistance (mortality rate: 90-93%) to deltamethrin and low resistance (mortality rate: 82-85%) to alphacypermethrin insecticides were detected. knockdown was more rapid with deltamethrin and alphacypermethrin than with permethrin across the study sites considering their KDT50 and KDT95. The frequency of the resistant phenotypes ranged from 35.14 to 55.3% across the study sites with a net of 45.1% resistant phenotype recorded in this study. The 1014F allelic frequency calculated from Hardy-Weinberg principle for vector populations across the study sites ranged from 0.500 (50.00%) to 0.7763 (77.63%). All populations witnessed significant (p < 0.05) deviations from Hardy-Weinberg equilibrium in the distribution of these alleles. The findings of this study show that there is a tendency to record an entire population of resistant vectors in this study area over time due to natural selection. The public health implication of these findings is that the use of pyrethroid based aerosols, coils, sprays, LLITNs and others for the purpose of controlling vectors of lymphatic filariasis and other diseases may be effort in futility.

Prevalence of permethrin-resistant kdr mutation in head lice (Pediculus humanus capitis) from elementary school students in Jeddah, Saudi Arabia.

Head lice (Pediculus humanus capitis) are a major global concern, and there is growing evidence of an increase in head lice prevalence among Saudi schoolchildren. The purpose of this study is to investigate the prevalence of an insecticidal resistance mutation in head lice collected from schoolchildren. A polymerase chain reaction (PCR) was used to amplify a segment of the voltage-gated sodium channel gene subunit to assess the prevalence and distribution of the kdr T917I mutation in head lice. Subsequently, the restriction fragment length polymorphism (RFLP) patterns revealed two genotypic forms: homozygous-susceptible (SS) and homozygous-resistant (RR). The results showed that 17 (37.80%) of the 45 samples were SS, whereas 28 (62.2%) were RR and T917I and L920F point mutations were found in the nucleotide and amino acid sequences of RR. Compared to other nations, the frequency of permethrin resistance mutation in the head louse population in Saudi Arabia was low. This study provides the first evidence of permethrin resistance mutation in human head lice in Saudi Arabia. The findings of this study will highlight the rising incidence of the kdr mutation in head lice in Saudi Arabia.

Susceptibility status of Aedes aegypti (Diptera: Culicidae) against insecticides of public health use in Delhi and NCR region, India.

BACKGROUND & OBJECTIVES: Aedes (Stegomyia) aegypti is a primary vector responsible for the transmission of various arboviral diseases in India. Without an effective drug or vaccine against these diseases, chemical insecticide-based vector control supplemented with source reduction remains the best option for their effective management. The development of insecticide resistance due to the continuous use of insecticides might affect the control operations. METHODS: Adults and larvae of Aedes aegypti were collected from different localities in Delhi. Larvae were exposed to discriminating (0.02mg/l) and application (1mg/l) doses of temephos. WHO tube assay was conducted for F1 adults using impregnated insecticide papers of dichlorodiphenyltrichloroethane (DDT), malathion, deltamethrin, permethrin, cyfluthrin, and lambda-cyhalothrin. RESULTS: Larvae of Ae. aegypti were found resistant (76.0%) to the discriminating dose of temephos, whereas suscep-tible (100.0%) to the application dose of the temephos. Adult Aedes (Fl) mosquitoes were resistant to DDT (23.7%), malathion (90.5%), deltamethrin (76.0%), permethrin (96.2 %) cyfluthrin (85.5%), and lambda-cyhalothrin (94.0%). INTERPRETATION & CONCLUSION: Indoor residual spray is not used in Delhi for vector control. Resistance in Aedes might be due to pesticide usage for agricultural activities in peripheral regions of Delhi. There is a need to investigate more on the insecticide resistance mechanisms for indirect resistance development. Understanding the insecticide susceptibility status of urban vectors is critical for planning effective control strategies.

Overexpression of cytochrome P450 and esterase genes involved in permethrin resistance in larvae and adults of Culex quinquefasciatus.

Mosquitoes are important vectors of several arthropod-borne diseases, which remain a priority for epidemiological research. Mosquito vector control strategies have traditionally relied on chemical insecticides such as synthetic pyrethroids. However, the indiscriminate use of pesticides has resulted in the development of resistance in many mosquito species. In insects, resistance evolves primarily through the overexpression of one or more gene products from the cytochrome P450, carboxylesterase, and glutathione superfamilies. The current study examined the expression of cytochrome P450 CYP6M2, CYP6AA7, CYP6Z2, CYP9J34, α-Esterase, Esterase B1, and neuroactin genes in larvae and adults of a permethrin-resistant (PerRes) and susceptible (Sus) Culex quinquefasciatus strains. The results showed that the CYP6AA7 gene was overexpressed (10-fold) in larvae and adults with PerRes (p < 0.01) followed by CYPJ34 (9.0-fold) and CYP6Z2 (5.0-fold) compared to the Sus, whereas fewer changes in CYP6M gene expression were observed in PerRes adults (p < 0.05), and no expression was found in larvae. The esterase gene was overexpressed in PerRes larvae (9.0-fold) followed by adults (2.5-fold) compared to the susceptible strain. Based on data, the present study suggests that cytochrome P450, CYP6AA7, CYP6Z2, CYP9J34, α-Esterase, Esterase B1, and neuroactin genes were involved in permethrin resistance in larval and adult Cx. quinquefasciatus.

Quantitative proteomics analysis of permethrin and temephos-resistant Ae. aegypti revealed diverse differentially expressed proteins associated with insecticide resistance from Penang Island, Malaysia.

Synthetic insecticides are the primary vector control method used globally. However, the widespread use of insecticides is a major cause of insecticide-resistance in mosquitoes. Hence, this study aimed at elucidating permethrin and temephos-resistant protein expression profiles in Ae. aegypti using quantitative proteomics. In this study, we evaluated the susceptibility of Ae. aegypti from Penang Island dengue hotspot and non-hotspot against 0.75% permethrin and 31.25 mg/l temephos using WHO bioassay method. Protein extracts from the mosquitoes were then analysed using LC-ESI-MS/MS for protein identification and quantification via label-free quantitative proteomics (LFQ). Next, Perseus 1.6.14.0 statistical software was used to perform differential protein expression analysis using ANOVA and Student's t-test. The t-test selected proteins with≥2.0-fold change (FC) and ≥2 unique peptides for gene expression validation via qPCR. Finally, STRING software was used for functional ontology enrichment and protein-protein interactions (PPI). The WHO bioassay showed resistance with 28% and 53% mortalities in adult mosquitoes exposed to permethrin from the hotspot and non-hotspot areas. Meanwhile, the susceptibility of Ae. aegypti larvae revealed high resistance to temephos in hotspot and non-hotspot regions with 80% and 91% mortalities. The LFQ analyses revealed 501 and 557 (q-value <0.05) differentially expressed proteins in adults and larvae Ae. aegypti. The t-test showed 114 upregulated and 74 downregulated proteins in adult resistant versus laboratory strains exposed to permethrin. Meanwhile, 13 upregulated and 105 downregulated proteins were observed in larvae resistant versus laboratory strains exposed to temephos. The t-test revealed the upregulation of sodium/potassium-dependent ATPase ß2 in adult permethrin resistant strain, H15 domain-containing protein, 60S ribosomal protein, and PB protein in larvae temephos resistant strain. The downregulation of troponin I, enolase phosphatase E1, glucosidase 2ß was observed in adult permethrin resistant strain and tubulin ß chain in larvae temephos resistant strain. Furthermore, the gene expression by qPCR revealed similar gene expression patterns in the above eight differentially expressed proteins. The PPI of differentially expressed proteins showed a p-value at <1.0 x 10-16 in permethrin and temephos resistant Ae. aegypti. Significantly enriched pathways in differentially expressed proteins revealed metabolic pathways, oxidative phosphorylation, carbon metabolism, biosynthesis of amino acids, glycolysis, and citrate cycle. In conclusion, this study has shown differentially expressed proteins and highlighted upregulated and downregulated proteins associated with insecticide resistance in Ae. aegypti. The validated differentially expressed proteins merit further investigation as a potential protein marker to monitor and predict insecticide resistance in field Ae. aegypti. The LC-MS/MS data were submitted into the MASSIVE database with identifier no: MSV000089259.

MiR-96-5p is involved in permethrin-promoted proliferation and migration of breast cancer cells.

MicroRNAs (miRNAs) are major players in cellular responses to xenobiotic compounds and toxins. However, the role of miRNAs in pyrethroid pesticide-induced cancer progression remains unclear. This study aimed to investigate the function of miR-96-5p in permethrin-induced proliferation and migration in breast cancer cells. In our study, the expression of miR-96-5p was upregulated in permethrin-treated MCF-7 cells. MiR-96-5p promoted MCF-7 cell proliferation and migration, accompanied bychanges in the expression of proteins involved in cell proliferation, migration, and apoptosis. Homeobox A5 (HOXA5) was identified as a direct target of miR-96-5p. HOXA5 silencing had the opposite effects with miR-96-5p inhibition. In conclusion, these results suggest that miR-96-5p is involved in permethrin-promoted proliferation and migration of breast cancer cells by targeting HOXA5.

Effects of next-generation, dual-active-ingredient, long-lasting insecticidal net deployment on insecticide resistance in malaria vectors in Tanzania: an analysis of a 3-year, cluster-randomised controlled trial.

BACKGROUND: Insecticide resistance among malaria-vector species is a pervasive problem that might jeopardise global disease-control efforts. Novel vector-control tools with different modes of action, including long-lasting insecticidal nets (LLINs) incorporating new active ingredients, are urgently needed to delay the evolution and spread of insecticide resistance. We aimed to measure phenotypic and genotypic insecticide-resistance profiles among wild Anopheles collected over 3 years to assess the longitudinal effects of dual-active-ingredient LLINs on insecticide resistance. METHODS: For this analysis, data nested in a 3-year, four parallel-arm, superiority cluster-randomised controlled trial (cRCT) in Tanzania, collected from 84 clusters (39 307 households) formed of 72 villages in the Misungwi district, were used to measure insecticide-resistance profiles among female Anopheles mosquitoes via insecticide-resistance bioassays and quantitative RT-PCR of metabolic-resistance genes. Wild, blood-fed, indoor-resting mosquitoes were collected annually during the rainy seasons from house walls in clusters from all four trial groups. Mosquitoes were morphologically identified as An gambiae sensu lato (SL) or An funestus SL before separate bioassay testing. The primary outcomes were lethal-dose values for α-cypermethrin, permethrin, and piperonyl butoxide pre-exposure plus permethrin-resistance intensity bioassays, mortality 72 h after insecticidal exposure for chlorfenapyr bioassays, fertility reduction 72 h after insecticidal exposure for pyriproxyfen bioassays, and fold change in metabolic-enzyme expression relative to an insecticide-susceptible laboratory strain. All primary outcomes were measured in An funestus SL 1 year, 2 years, and 3 years after LLIN distribution. Primary outcomes were also assessed in An gambiae SL if enough mosquitoes were collected. The cRCT is registered with ClinicalTrials.gov (NCT03554616). FINDINGS: Between May 24, 2019, and Oct 25, 2021, 47 224 female Anopheles were collected for resistance monitoring. In the pyrethroid (PY)-LLIN group, there were significant increases in α-cypermethrin-resistance intensity (year 1 LD50=9·52 vs year 2 76·20, p<0·0001) and permethrin-resistance intensity (year 1 13·27 vs year 2 35·83, p=0·0019) in An funestus SL. In the pyriproxyfen PY-LLIN group, there was similar increase in α-cypermethrin-resistance intensity (year 1 0·71 vs year 2 81·56, p<0·0001) and permethrin-resistance intensity (year 1 5·68 vs year 2 50·14, p<0·0001). In the piperonyl butoxide PY-LLIN group, α-cypermethrin-resistance intensity (year 1 33·26 vs year 3 70·22, p=0·0071) and permethrin-resistance intensity (year 1 47·09 vs year 3 2635·29, p<0·0001) also increased over time. In the chlorfenapyr PY-LLIN group, there were no effects on α-cypermethrin-resistance intensity (year 1 0·42 vs year 3 0·99, p=0·54) or permethrin-resistance intensity (data were not estimable due to nearly 100% mortality). There were also minimal reductions in chlorfenapyr susceptibility. However, in the chlorfenapyr PY-LLIN group, a significant decline in piperonyl-butoxide synergy was seen by year 3 (year 1 0·02 vs year 3 0·26, p=0·020). Highly over-expressed detoxification enzymes showed dynamic patterns of selection throughout the trial. INTERPRETATION: Our phenotypic data supports trial epidemiological findings; chlorfenapyr PY-LLINs provided superior protection from malaria across multiple transmission seasons, with few effects on insecticide-resistance selection. Rapid pyrethroid-resistance intensification in the piperonyl butoxide PY-LLIN group and pre-existing tolerance of pyriproxyfen in vector populations might explain the poorer performance of these two interventions regarding malaria outcomes. Further work is required to elucidate the potential mechanisms driving cross-resistance between pyrethroids and novel active ingredients to better inform the design of pre-emptive resistance-management strategies. FUNDING: UK Department for International Development; UK Medical Research Council; Wellcome Trust; UK Department of Health and Social Care; UK Foreign, Commonwealth and Development Office; and The Bill and Melinda Gates Foundation via the Innovative Vector Control Consortium.

Effects of permethrin on ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) in ornamental nurseries.

Exotic ambrosia beetles (Coleoptera: Curculionidae: Scolytinae), such as Xylosandrus crassiusculus (Motschulsky), Xylosandrus germanus (Blandford), and Xylosandrus compactus (Eichoff) are serious pests in southeastern ornamental nurseries. Preventative pyrethroid trunk sprays effectively reduce boring damage. However, it is unclear how pyrethroids such as permethrin prevent attack. Thus, the objective was to determine how permethrin-treated bolts interact with invading ambrosia beetles. In 2022, a study with 2 independent trials was conducted in a nursery on red maple (Acer rubrum L.), bolts during March and April, respectively. The treatments were (i) nonbaited, nontreated bolt, (ii) ethanol baited bolt, (iii) nonbaited bolt + glue [painted on bolt], (iv) ethanol baited bolt + glue, (v) ethanol baited bolt + glue + permethrin, (vi) ethanol baited bolt + glue + permethrin + verbenone, and (vii) ethanol baited bolt + glue + verbenone. Ambrosia beetles trapped on glue, beetles which fell into the pail with soap solution under the bolts, and entry holes on bolts were quantified. Permethrin prevented beetle attacks but did not reduce the number of ambrosia beetles landing on the treated bolts. Verbenone reduced ambrosia beetles from landing on the bolts but did not prevent boring into bolts. The numbers of ambrosia beetles in soapy water were not significantly different among treatments. Ambrosia beetles are landing on permethrin-treated bolts but not boring into the bolts, implying that fresh permethrin residues may not be necessary for ambrosia beetle management.

Pseudoresistance to permethrin in scabies.

Reports of permethrin resistance of Sarcoptes scabiei var. hominis are emerging. We propose that this may be pseudoresistance. The resistance is due to inadequate counselling by physicians, incorrect treatment (insufficient quantity of permethrin; too short length of treatment), and poor adherence and compliance by patients. Other reasons include single application of permethrin, suggestion to apply the drug for 6-8 hours, failed application on subungual folds, irritant contact dermatitis, in particular on genitals, for which some patients stop the treatment, and unexplainable use of permethrin in post-scabies prurigo. Thus, we believe that several cases of resistance to permethrin are actually cases of pseudoresistance.

Detection of a knockdown mutation in the voltage-sensitive sodium channel associated with permethrin tolerance in Sarcoptes scabiei var. hominis mites.

BACKGROUND: Increasing evidence has sparked a debate on the loss of sensitivity of scabies mites to conventional permethrin therapy. Mutations in the voltage-sensitive sodium channels (VSSC) were associated with knockdown resistance (kdr) in many arthropods, but have never been identified in Sarcoptes scabiei variatio (var.) hominis mites. OBJECTIVES: To identify factors contributing to therapy failure. METHODS: Sixty-seven mites were collected from 64 scabies-infested patients in Vienna, Austria, of whom 85.9% were refractory to prior permethrin-based treatments, and genotyped for the presence of nucleotide polymorphisms in Domain II of the VSSC, known to be associated with kdr. Information regarding previous antiscabietic therapies, decontamination procedures and possible re-infestations by contacts as well as the response to re-imposed therapies were obtained. RESULTS: Sequence alignment comparisons revealed previously unidentified mutations in the coding region of Domain II of the VSSC. A novel A1663T transversion was detected in 97.0% of the mites, resulting in a non-synonymous substitution from methionine to leucine, M918L, a mutation known to confer kdr in other arthropods. In addition, a synonymous G1659A transition was identified in one mite, which otherwise showed a nucleotide sequence identical to the wild-type reference. No major inconsistencies were observed within the previous therapeutic and decontamination procedures, which could have accounted for the observed non-responsiveness to permethrin-based therapies. Subsequent cure of infestation was achieved in 65.6% of the participants, predominantly by combination therapies with topical permethrin and systemic ivermectin. However, in 14.6% of the cured cases, permethrin monotherapy sufficed for eradication of scabies, albeit in some cases prolonged exposure was necessary. CONCLUSIONS: The kdr-associated M918L mutation in the VSSC gene has now emerged in S. scabiei var. hominis mites. Hence, loss of sensitivity to permethrin due to kdr-type resistance may be more prevalent than anticipated and may be decisive for the therapy responsiveness of scabies-infested patients.

Microbial Diversity and Enzyme Activity as Indicators of Permethrin-Exposed Soil Health.

Owing to their wide range of applications in the control of ticks and insects in horticulture, forestry, agriculture and food production, pyrethroids pose a significant threat to the environment, including a risk to human health. Hence, it is extremely important to gain a sound understanding of the response of plants and changes in the soil microbiome induced by permethrin. The purpose of this study has been to show the diversity of microorganisms, activity of soil enzymes and growth of Zea mays following the application of permethrin. This article presents the results of the identification of microorganisms with the NGS sequencing method, and of isolated colonies of microorganisms on selective microbiological substrates. Furthermore, the activity of several soil enzymes, such as dehydrogenases (Deh), urease (Ure), catalase (Cat), acid phosphatase (Pac), alkaline phosphatase (Pal), ß-glucosidase (Glu) and arylsulfatase (Aryl), as well as the growth of Zea mays and its greenness indicators (SPAD), after 60 days of growth following the application of permethrin, were presented. The research results indicate that permethrin does not have a negative effect on the growth of plants. The metagenomic studies showed that the application of permethrin increases the abundance of Proteobacteria, but decreases the counts of Actinobacteria and Ascomycota. The application of permethrin raised to the highest degree the abundance of bacteria of the genera Cellulomonas, Kaistobacter, Pseudomonas, Rhodanobacter and fungi of the genera Penicillium, Humicola, Iodophanus, Meyerozyma. It has been determined that permethrin stimulates the multiplication of organotrophic bacteria and actinomycetes, decreases the counts of fungi and depresses the activity of all soil enzymes in unseeded soil. Zea mays is able to mitigate the effect of permethrin and can therefore be used as an effective phytoremediation plant.