Permethrin Resistance Research Articles

Unveiling the Role of Two Rhodopsin-like GPCR Genes in Insecticide-Resistant House Flies, Musca domestica

Insecticide resistance in insects, driven by the overexpression of P450 enzymes, presents a significant challenge due to the enhanced metabolic detoxification of insecticides. Although the transcriptional regulation of P450 genes is not yet fully understood, G-protein-coupled receptor (GPCR) genes have emerged as key regulators in this process. This study is the first to associate GPCR genes with insecticide resistance in Musca domestica. We identified two key rhodopsin-like GPCR genes, ALHF_02706.g1581 and ALHF_04422.g2918, which were significantly overexpressed in the resistant ALHF strain compared to sensitive strains. Notably, both ALHF_02706.g1581 and ALHF_04422.g2918 were mapped to autosome 2, where critical but unidentified regulatory factors controlling resistance and P450 gene regulation are located. This supports our hypothesis that GPCRs function as trans-regulatory factors for P450-mediated resistance. Functional analysis using transgenic Drosophila demonstrated that overexpression of these rhodopsin-like GPCR genes increased permethrin resistance by approximately two-fold. Specifically, ALHF_02706.g1581 overexpression significantly upregulated the Drosophila resistance-related P450 genes CYP12D1, CYP6A2, and CYP6A8, while ALHF_04422.g2918 increased CYP6G1 and CYP6A2 expression, thereby enhancing insecticide detoxification in rhodopsin-like GPCR transgenic Drosophila lines. These findings suggest that these rhodopsin-like GPCR genes on autosome 2 may act as trans-regulatory factors for P450-mediated resistance, underscoring their critical role in insecticide detoxification and resistance development in M. domestica.

A severe head louse (Pediculus humanus capitis) infestation: Timeline, pathogen detection, kdr-gene screening and morphological anomalies of lice.

Lice are assuming an increasing importance in forensic investigations, given their capacity to provide information about an individual's care. Head louse pediculosis is a frequent condition in school-age children and can be properly controlled using topical treatments combined with good personal hygiene. Prolonged and chronic infestations may result in more serious outcomes including severe iron deficiency anaemia. We conducted entomological and laboratory investigations of a head louse infestation in a 12-year-old girl who experienced severe anaemia and subsequent death. Numerous lice were found postmortem on the head, face and neck of the patient, as well as on bedding and clothing. Analysis of nits on individual hairs determined that the louse infestation had been present for at least 166 days. The lice had some morphological traits characteristic of body lice: the third antennal segment in some specimens was distinctly longer than wide, and the apices of some paratergal plates did not extend into intersegmental membranes, while other morphological features were characteristic of head or body lice. All lice were heterozygous for the T917I kdr genotype, a marker of permethrin resistance. Nineteen (79.2%, 95%CI 59.5%-90.8%) louse DNA samples tested TaqMan positive for Acinetobacter (Moraxellales; Moraxellaceae) sp. Available information and laboratory findings are further discussed regarding their possible contribution to the negative outcome of this case. We stress the impact head louse pediculosis can have on children with limited parental attention, and how severe head louse infestation may serve as warning sign of neglect, and other high-risk situations.

Mutational Study of Voltage-Gated Sodium Channel in Permethrin Resistance in Mosquitoes: An in silico Study

Mutational Study of Voltage-Gated Sodium Channel in Permethrin Resistance in Mosquitoes: An in silico Study

First report of V1016I, F1534C and V410L kdr mutations associated with pyrethroid resistance in Aedes aegypti populations from Niamey, Niger.

Ae. aegypti is the vector of important μ arboviruses, including dengue, Zika, chikungunya and yellow fever. Despite not being specifically targeted by insecticide-based control programs in West Africa, resistance to insecticides in Ae. aegypti has been reported in countries within this region. In this study, we investigated the status and mechanisms of Ae. aegypti resistance in Niamey, the capital of Niger. This research aims to provide baseline data necessary for arbovirus outbreak prevention and preparedness in the country. Ovitraps were used to collect Ae. aegypti eggs, which were subsequently hatched in the insectary for bioassay tests. The hatched larvae were then reared to 3-5-day-old adults for WHO tube and CDC bottle bioassays, including synergist tests. The kdr mutations F1534C, V1016I, and V410L were genotyped using allele-specific PCR and TaqMan qPCR methods. Ae. aegypti from Niamey exhibited moderate resistance to pyrethroids but susceptibility to organophosphates and carbamates. The kdr mutations, F1534C, V1016I and V410L were detected with the resistant tri-locus haplotype 1534C+1016L+410L associated with both permethrin and deltamethrin resistance. Whereas the homozygote tri-locus resistant genotype 1534CC+1016LL+410LL was linked only to permethrin resistance. The involvement of oxidase and esterase enzymes in resistance mechanisms was suggested by partial restoration of mosquitoes' susceptibility to pyrethroids in synergist bioassays. This study is the first report of Ae. aegypti resistance to pyrethroid insecticides in Niamey. The resistance is underpinned by target site mutations and potentially involves metabolic enzymes. The observed resistance to pyrethroids coupled with susceptibility to other insecticides, provides data to support evidence-based decision-making for Ae. aegypti control in Niger.

Insecticide resistance level of Aedes aegypti L. (Diptera: Culicidae) in northern Mexico

Objective: To determine the response of the larvae of three Ae. aegypti populations from the Mexican North Pacific region to insecticides with different mode of action.
 Design/Methodology/Approach: Three colonies were obtained placing ovitramps in peridomestic sites in Guadalajara (Jalisco), Culiacan (Sinaloa), and La Paz (Baja California Sur). Based on the methodology proposed by WHO, the bioassays were carried out with F1 larvae in the early fourth instar.
 Results: The larvae from the three field colonies had high resistance to permethrin and low resistance to deltamethrin; however, they were susceptible to Spinosad and Bacillus thuringiensis var. Israelensis. The Culiacan strain showed a high resistance to the malathion and propoxur insecticides.
 Study Limitations/Implications: The results provided valuable information about the response of these populations to insecticides, which are useful to establish resistance in the lab. Consequently, further studies should be carried out to complement the information obtained in these field tests.
 Findings/Conclusions: The data indicated resistance levels to pyrethroid insecticides (mainly permethrin), as well as to organophosphates and carbamates.

Implications from an Ongoing Scabies Outbreak During the COVID-19 Isolation Period: Clues for Controlling Scabies Today

Background: Contrary to the expectation that the number of scabies cases will decrease during the COVID-19 isolation period, scabies has been frequently reported in Turkey. 
 Objective: The aim of this study is to investigate the epidemiological pattern of the scabies outbreak in the context of the pandemic to provide additional information for the control of the devastating scabies outbreak that continues today.
 Methods: Patients seen in our dermatology clinic before and during the pandemic were evaluated retrospectively. 
 Results: There was no decrease in the rate of scabies patients among all dermatology admissions during the pandemic period (n=143/2912; 4.9%) compared to the pre-pandemic period (n=526/11679; 4.5%) (p=0.348). The high incidence of the new scabies cases before the detection of COVID-19 in the country decreased in the quarantine period. However, the high recurrent admissions of old scabies cases, who already got scabies before pandemic but could not be cured despite repeated treatments, contributed to the ongoing high prevalance of scabies during the pandemic (one-third of all scabies cases seen during the pandemic, n=30/87).
 Conclusions: We suggest that self-quarantine and increased hygiene during COVID-19 reduced re-infestations and led to a decrease in new cases. Treatment failure of old cases has been observed to contribute to the high prevalence of scabies in the pandemic. Permethrin resistance may have led to inadequate treatment success, given that treatment application errors, which may also lead to treatment failure, are managed with active surveillance in our patient population. Studies addressing permethrin resistance are needed as a priority.

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.

Study of Resistance of Aedes aegypti Mosquitoes to Pyrethroids by Free-flight Method

Efforts to combat dengue hemorrhagic fever (DHF) in Bali are focused on controlling the vector of Aedes aegypti Mosquitoes. One method is fumigation (fogging) with insecticides. The use of insecticides for a long time and improper doses can trigger resistance. Many countries including Indonesia have reported incidents of insecticide resistance. Methods for determining the level of insecticide resistance are bioassay, biochemistry, and gene mutation. The method does not describe the actual environmental conditions of Mosquitoes. Queensland Institute of Medical Research (QIMR) Berghofer developed a free-flight test procedure to evaluate insecticide resistance in free-flying mosquito conditions. The study, which was conducted in June 2019-January 2020, aimed to measure the level of insecticide resistance with the free-flight method in the A. aegypti population in North Denpasar. A total of 20 female A. aegypti aged 5-10 days were released in the free-flight room for 24 hours with permethrin or cypermethrin insecticide boards. The level of resistance is determined based on the mosquito mortality rate using WHO standards. The results showed permethrin and cypermethrin resistance in the A. aegypti population of North Denpasar, with mortality of less than 80%.

Genetics, genomics and mechanisms responsible for high levels of pyrethroid resistance in Musca domestica

Insecticide resistance is both economically important and evolutionarily interesting phenomenon. Identification of the mutations responsible for resistance allows for highly sensitive resistance monitoring and allows tools to study the forces (population genetics, fitness costs, etc.) that shape the evolution of resistance. Genes coding for insecticide targets have many well-characterized mutations, but the mutations responsible for enhanced detoxification have proven difficult to identify.We employed multiple strategies to identify the mutations responsible for the extraordinarily high permethrin resistance in the KS17-R strain of house fly (Musca domestica): insecticide synergist assays, linkage analysis, bulk segregant analyses (BSA), transcriptomics and long read DNA (Nanopore) sequencing. The >85,100-fold resistance in KS17-R was partially suppressed by the insecticide synergists piperonyl butoxide and S,S,S-tributylphosphorothionate, but not by diethyl maleate nor by injection. This suggests the involvement of target site insensitivity, CYP-mediated resistance, possibly hydrolase mediated resistance and potentially other unknown factors. Linkage analysis identified chromosomes 1, 2, 3 and 5 as having a role in resistance. BSA mapped resistance loci on chromosomes 3 and 5. The locus on chromosome 3 was centered on the voltage sensitive sodium channel. The locus on chromosome 5 was associated with a duplication of multiple detoxification genes. Transcriptomic analyses and long read DNA sequencing revealed overexpressed CYPs and esterases and identified a complex set of structural variants at the chromosome 5 locus.

National survey in Switzerland calls for improved diagnosis and treatment in children with scabies.

The global prevalence of scabies is estimated to be up to 200 million cases annually, with young children particularly affected. In Europe, most cases are thought to originate in migrant populations. Scabies management is challenging in children. To identify knowledge gaps and research needs, we aimed to descriptively evaluate the management of children with scabies by different Swiss healthcare providers. An invitation for an anonymous online survey (36 questions) was sent to members of Swiss societies of dermatologists, general practitioners, paediatricians, paediatric dermatologists, paediatric infectious diseases specialists, and tropical medicine specialists, inviting clinicians to participate from 25thMay to 8thAugust 2020. One reminder invitation was sent. Hospital pharmacies and the distributor of permethrin were contacted to report consumption trends of scabicides in 2018 and 2019. The survey was completed by 248 clinicians: 146 (59%) paediatricians, 47 (19%) dermatologists, 28 (11%) general practitioners, 6 (2%) paediatric dermatologists, 13 (5%) paediatric infectious diseases specialists, and 8 (3%) tropical medicine specialists. Most consulted up to 10 scabies cases within a 16-month period, with similar numbers in migrant and Swiss children. Dermoscopy was used by 24% of non-dermatologists. Non-dermatologists did not consider co-treatment of close contacts in up to 59% of cases. While permethrin was the first-line treatment, treatment failures were frequently reported in children aged <5 years. Up to 67% of paediatric dermatologists regularly used oral ivermectin off-label in children weighing <15 kg. None of the paediatric dermatologists, 15% of the dermatologists, and 9% of the non-dermatologists used only one treatment cycle.Scabicide consumption increased. Treatment studies on ivermectin use in children weighing <15 kg had the highest research priority. In Switzerland, scabies is a frequent dermatosis in migrant and Swiss children. While accessible, optimal diagnostics are underutilised, and treatment is suboptimal. Permethrin resistance appears to be an increasing problem. Dermatologists regularly use ivermectin off-label in children weighing <15 kg. Treatment studies on ivermectin use in children weighing <15 kg, user-friendly diagnostic tools, new treatment protocols, and child-friendly dosage forms are needed to improve the diagnosis and treatment of children with scabies.

Correlation between pyrethroid knockdown resistance and mutation frequency of voltage-gated sodium channel and its application in Aedes aegypti management

Aedes aegypti, the primary vector responsible for transmitting dengue fever in southern Taiwan, has developed a relatively high resistance to synthetic pyrethroids. It has evolved four amino acid substitutions in the voltage-gated sodium channel (VGSC), namely S996P, V1023G, F1565C, and D1794Y. To unveil the distribution and correlation of VGSC mutations and pyrethroid resistance among different field populations, Ae. aegypti collected from various districts in Kaohsiung and Tainan Cities underwent tests for resistance development against different pyrethroids and frequency of S996P, V1023G, F1565C, and D1794Y substitutions. The adult knockdown assay revealed a relatively high knockdown resistance in the Ae. aegypti populations from Kaohsiung and Tainan against permethrin, cypermethrin, and fenvalerate (averaging >50-fold). Conversely, less resistance was observed against α-cypermethrin, deltamethrin, λ-cyhalothrin, cyfluthrin, and etofenprox (averaging <35-fold). Using Polymerase Chain Reaction/restriction fragment length polymorphism analysis, four mutant haplotypes were identified in these field populations. Notably, the SIAVFD and SIBVFD wild haplotypes were absent. Analysis utilizing IBM SPSS Statistics 20.0 and Spearman's rank correlation coefficient indicated that Haplotype C (PIAGFD), especially P allele, frequency displayed a significant positive correlation with five Type II pyrethroid resistance, while 1023G and 1023G/G exhibited a significant association with permethrin and fevalerate resistance. Conversely, Haplotype E (SIBVCD) negatively correlated with pyrethroid resistance, particularly fenvalerate resistance (−0.776). Haplotype C and E were the most prevalent and widely distributed among the investigated field populations. This prevalence of haplotype C is likely tied to the extensive and excessive use of Type II pyrethroids for dengue control over the past three decades. Given the significant positive correlation, the best-fit lines and R2 values were established to facilitate the swift prediction of knockdown resistance levels to various pyrethroids based on VGSC mutation frequency. This predictive approach aims to guide insecticide usage and the management of pyrethroid resistance in the field populations of Ae. aegypti in Taiwan.

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.

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.

Resistance risk assessment of six pyrethroids and acephate toward the resistant adult tarnished plant bug, Lygus lineolaris.

Due to rapidly developed resistance, pest management relies less on pyrethroids to control economically damaging infestations of the tarnished plant bug (TPB), Lygus lineolaris (Palisot de Beauvois) in cotton fields of Mississippi. Yet, pyrethroid resistance remains prevalent in TPB populations. This study assessed the resistance levels in adult TPB to six common pyrethroids and acephate. Resistant TBPs were collected from wild host plants in late October after harvest in the Mississippi Delta region of the United States. Based on LC50 values, the field-resistant TPBs displayed higher resistance to permethrin, esfenvalerate, and bifenthrin (approximately 30 fold) and moderate resistance to λ-cyhalothrin, β-cyfluthrin, ζ-cypermethrin, and acephate (approximately 15 fold). Further investigations showed that the inhibitors of three detoxification enzyme, triphenyl phosphate (TPP), diethyl maleate (DEM), and piperonyl butoxide (PBO) had synergistic effects on permethrin, λ-cyhalothrin, and bifenthrin in resistant TPBs. Furthermore, elevated esterase, GST, and P450 activities were significantly expressed in field-resistant TPBs. Additionally, GST and esterase were reduced after 48 h exposure to certain pyrethroids at LC50 dose. The synergistic and biochemical assays consistently indicated that P450 and esterase were involved in pyrethroid detoxification in TPBs. This study provides valuable information for the continued use of pyrethroids and acephate in controlling TPBs in cotton fields in the Mississippi Delta region of the United States.

Permethrin-unresponsive scabies in London, United Kingdom - a wake up call.

Failure to respond to permethrin, the most widely prescribed first-line therapy for scabies, has been increasingly reported in recent years in a number of countries. However, its true scale and relation to verifiable permethrin resistance is currently unknown. We present a case of scabies associated with scabetic nodules in which a lack of response to topical permethrin treatment was evident.

Toxicity, resistance and metabolic mechanism of resistance to permethrin in larvae of laboratory and field strains of Musca domestica L.

Management of Musca domestica is desirable globally in order to manage public health issues. However, development of insecticide resistance is one of major hindrances in the management of M. domestica. Here, we explored toxicity of permethrin against larvae of laboratory and field strains of M. domestica, resistance comparison, synergistic effect of enzyme inhibitors on permethrin toxicity, and activities of metabolic enzymes in both strains. In case of the Lab-susceptible strain, LC50 values of permethrin were 0.14, 0.52 and 0.94 µg g-1 for first, second and third instars, respectively. In case of the field strain, LC50 values were 28.09, 66.86 and 112.24 µg g-1 for first, second and third instars, respectively. First, second and third instars of the field strain exhibited 200.64, 128.58 and 119.40 fold resistance to permethrin when compared with the first, second and third instars of the Lab-susceptible strain, respectively. Toxicity of permethrin in the field strain synergized in the presence of enzyme inhibitors (PBO or DEF). Enzyme analysis revealed significantly higher activities of MFO, CarE and GST in the field strain than those of the Lab-susceptible strain, suggesting metabolic-based mechanism of permethrin resistance. In conclusion, the field strain exhibited resistance to permethrin, which could be managed by adopting alternate measures for the management of M. domestica.

Molecular drivers of insecticide resistance in the Sahelo-Sudanian populations of a major malaria vector Anopheles coluzzii

BackgroundInformation on common markers of metabolic resistance in malaria vectors from countries sharing similar eco-climatic characteristics can facilitate coordination of malaria control. Here, we characterized populations of the major malaria vector Anopheles coluzzii from Sahel region, spanning four sub-Saharan African countries: Nigeria, Niger, Chad and Cameroon.ResultsGenome-wide transcriptional analysis identified major genes previously implicated in pyrethroid and/or cross-resistance to other insecticides, overexpressed across the Sahel, including CYP450s, glutathione S-transferases, carboxylesterases and cuticular proteins. Several, well-known markers of insecticide resistance were found in high frequencies—including in the voltage-gated sodium channel (V402L, I940T, L995F, I1527T and N1570Y), the acetylcholinesterase-1 gene (G280S) and the CYP4J5-L43F (which is fixed). High frequencies of the epidemiologically important chromosomal inversion polymorphisms, 2La, 2Rb and 2Rc, were observed (~80% for 2Rb and 2Rc). The 2La alternative arrangement is fixed across the Sahel. Low frequencies of these inversions (<10%) were observed in the fully insecticide susceptible laboratory colony of An. coluzzii (Ngoussou). Several of the most commonly overexpressed metabolic resistance genes sit in these three inversions. Two commonly overexpressed genes, GSTe2 and CYP6Z2, were functionally validated. Transgenic Drosophila melanogaster flies expressing GSTe2 exhibited extremely high DDT and permethrin resistance (mortalities <10% in 24h). Serial deletion of the 5′ intergenic region, to identify putative nucleotide(s) associated with GSTe2 overexpression, revealed that simultaneous insertion of adenine nucleotide and a transition (T->C), between Forkhead box L1 and c-EST putative binding sites, were responsible for the high overexpression of GSTe2 in the resistant mosquitoes. Transgenic flies expressing CYP6Z2 exhibited marginal resistance towards 3-phenoxybenzylalcohol (a primary product of pyrethroid hydrolysis by carboxylesterases) and a type II pyrethroid, α-cypermethrin. However, significantly higher mortalities were observed in CYP6Z2 transgenic flies compared with controls, on exposure to the neonicotinoid, clothianidin. This suggests a possible bioactivation of clothianidin into a toxic intermediate, which may make it an ideal insecticide against populations of An. coluzzii overexpressing this P450.ConclusionsThese findings will facilitate regional collaborations within the Sahel region and refine implementation strategies through re-focusing interventions, improving evidence-based, cross-border policies towards local and regional malaria pre-elimination.

Association of 410L, 1016I and 1534C kdr mutations with pyrethroid resistance in Aedes aegypti from Ouagadougou, Burkina Faso, and development of a one-step multiplex PCR method for the simultaneous detection of 1534C and 1016I kdr mutations

BackgroundSince 2000, Burkina Faso has experienced regular dengue cases and outbreaks, making dengue an increasingly important health concern for the country. Previous studies in Burkina Faso reported that resistance of Aedes aegypti to pyrethroid insecticides was associated with the F1534C and V1016I kdr mutations. The current study reports high resistance of Ae. aegypti populations to pyrethroid insecticides, likely supported by mutations in the voltage-gated sodium channel, here evidenced by genotyping the kdr SNPs V410L, V1016I and F1534C. We also describe a new multiplex PCR-based diagnostic of F1534C and V1016I kdr SNPs.MethodsLarvae of Ae. aegypti were collected from three health districts of Ouagadougou in 2018. The resistance status of Ae. aegypti to permethrin (15 μg/ml) and deltamethrin (10 μg/ml) was tested using bottles and to malathion (5%) using WHO tube tests. All bioassays used 1-h exposure and mortality recorded 24 h post-exposure. Bioassay results were interpreted according to WHO thresholds for resistance diagnosis. The kdr mutations were screened using AS-PCR and TaqMan methods in exposed and non-exposed Aedes mosquitoes.ResultsFemales from all health districts were resistant to permethrin and deltamethrin (< 20% mortality) but were fully susceptible to 5% malathion. The F1534C and V1016I kdr mutations were successfully detected using a newly developed multiplex PCR in perfect agreement with TaqMan method. The 1534C/1016I/410L haplotype was correlated with permethrin resistance but not with deltamethrin resistance; however, the test power was limited by a low frequency of dead individuals in deltamethrin exposure.ConclusionsResistance to pyrethroid insecticides is associated with kdr mutant haplotypes, while the absence of substantial resistance to malathion suggests that it remains a viable option for dengue vector control in Ouagadougou.

Contrasting Patterns of Asaia Association with Pyrethroid Resistance Escalation between the Malaria Vectors Anopheles funestus and Anopheles gambiae

Microbiome composition has been associated with insecticide resistance in malaria vectors. However, the contribution of major symbionts to the increasingly reported resistance escalation remains unclear. This study explores the possible association of a specific endosymbiont, Asaia spp., with elevated levels of pyrethroid resistance driven by cytochrome P450s enzymes and voltage-gated sodium channel mutations in Anopheles funestus and Anopheles gambiae. Molecular assays were used to detect the symbiont and resistance markers (CYP6P9a/b, 6.5 kb, L1014F, and N1575Y). Overall, genotyping of key mutations revealed an association with the resistance phenotype. The prevalence of Asaia spp. in the FUMOZ_X_FANG strain was associated with the resistance phenotype at a 5X dose of deltamethrin (OR = 25.7; p = 0.002). Mosquitoes with the resistant allele for the markers tested were significantly more infected with Asaia compared to those possessing the susceptible allele. Furthermore, the abundance correlated with the resistance phenotype at 1X concentration of deltamethrin (p = 0.02, Mann-Whitney test). However, for the MANGOUM_X_KISUMU strain, findings rather revealed an association between Asaia load and the susceptible phenotype (p = 0.04, Mann-Whitney test), demonstrating a negative link between the symbiont and permethrin resistance. These bacteria should be further investigated to establish its interactions with other resistance mechanisms and cross-resistance with other insecticide classes.

Multiple-P450 Gene Co-Up-Regulation in the Development of Permethrin Resistance in the House Fly, Musca domestica

This paper reports a study conducted at the whole transcriptome level to characterize the P450 genes involved in the development of pyrethroid resistance, utilizing expression profile analyses of 86 cytochrome P450 genes in house fly strains with different levels of resistance to pyrethroids/permethrin. Interactions among the up-regulated P450 genes and possible regulatory factors in different autosomes were examined in house fly lines with different combinations of autosomes from a resistant house fly strain, ALHF. Eleven P450 genes that were significantly up-regulated, with levels > 2-fold those in the resistant ALHF house flies, were in CYP families 4 and 6 and located on autosomes 1, 3 and 5. The expression of these P450 genes was regulated by trans- and/or cis-acting factors, especially on autosomes 1 and 2. An in vivo functional study indicated that the up-regulated P450 genes also conferred permethrin resistance in Drosophila melanogaster transgenic lines. An in vitro functional study confirmed that the up-regulated P450 genes are able to metabolize not only cis- and trans-permethrin, but also two metabolites of permethrin, PBalc and PBald. In silico homology modeling and the molecular docking methodology further support the metabolic capacity of these P450s for permethrin and substrates. Taken together, the findings of this study highlight the important function of multi-up-regulated P450 genes in the development of insecticide resistance in house flies.