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Head lice infestation (HLI), caused by Pediculus humanus capitis De Geer, 1767, has long been a common global problem of school children. Permethrin is an old pyrethroid derivative that has been used commonly for its treatment, and it exerts its activity over the voltage-sensitive calcium channels (VSCC) of the lice. There has been a growing list of persistent HLI cases lately in the world among patients using permethrin, and knockdown resistance (kdr)-related point mutations on VSCC have been identified and reported from those resistant lice samples. The aim of this study was to investigate the gene mutations associated with permethrin resistance in head lice collected from primary school children in Istanbul (Türkiye) and Nagarkot (Nepal) for the first time. A total of 192 P. h. capitis adults were collected from school children aged 6-12 years in two cities (96 lice each). Following DNA isolation, the fragment of the VSCC a-subunit gene, which contained the possible mutation sites ((kdr-like M815I (ATG > ATT), kdr T917I (ACA > ATA), and kdr-like L920F (CTT > TTT)), was amplified in each louse by PCR, and the PCR products were sequenced and aligned, followed by frequency calculations for alleles, genotypes, and haplotypes. Using nucleic acid sequence analysis, it was revealed that M815I, T917I, or L920F mutations were present on the VSCC genes in the lice samples from both Türkiye and Nepal. In addition, genotypic analyses indicated the presence of all three mutations in the lice samples from Türkiye, while the T917I mutation was detected in none of the lice collected in Nepal. This is the first report of gene mutations associated with permethrin resistance in head lice collected from a group of primary school children in the largest city of Türkiye (Istanbul) and Nagarkot. High mutation rates were identified in the lice, especially those from Istanbul, which is concordant with our previous unpublished study, in which almost 60% of the examined lice of the school children (in the same school selected in this study) remained alive despite long-term exposure to permethrin in the laboratory. These initial results show that gene mutations associated with permethrin resistance are common in lice samples in Istanbul and Nagarkot, which may suggest the current need for the selection of new pediculicidal agents in HLI treatment.

This study investigates the effect of mosquito sex and age on the survival and resistance determination of adult Culex tarsalis exposed to permethrin, a pyrethroid commonly used for mosquito control, using the Centers for Disease Control and Prevention (CDC) bottle bioassay method. A permethrin-susceptible strain of Cx. tarsalis (Bakersfield strain) was used in this study. Survival was compared for young adult females (2-5 days old) relative to older adult females (7-10 days old) and separately for female and male mosquitoes of the same age (2-5 days old). Mortality was slightly higher for males than females during the first observation period (0-5 min) following permethrin exposure, and higher for older females relative to younger females from 5 to 10 min following permethrin exposure, with no differences in mortality by either sex or age for observation periods during the remainder of the diagnostic period. When evaluated over the full diagnostic period, survival varied with mosquito age but not sex. However, all mosquitoes, regardless of sex or age, died within the 30-min diagnostic period for this species, confirming their permethrin susceptibility per the CDC bottle bioassay. This research contributes valuable insight into the potential impact of sex and age on mosquito susceptibility to insecticides in the context of insecticide resistance determination., (Copyright © 2024 by The American Mosquito Control Association, Inc.)

Pyrethroids are pesticides used in agriculture, the textile industry, wood processing, and human and animal medicine. Pyrethroids inhibit voltage-sensitive sodium channels (VSSCs) in insects and mammals. It results in the premature opening and/or delayed closing of the channels, causing a prolonged influx of Na + ions into the cell. Insects absorb pyrethroids throughout the entire body surface, while poisoning in humans most often occurs by inhalation and through the skin. In this study, 52 fragments of human skin taken from the eyelid fold were examined. A modified Ussing chamber was used to measure the active ion transport in epithelial tissue and quantify the tissue viability and integrity. Both permethrin and deltamethrin solutions induced changes in the transport of ions, mainly sodium, but by different mechanisms. Permethrin affected the transepithelial transport of sodium ions in a long-term mechanism, while deltamethrin affected the ability to respond to stimuli in an immediate mechanism. Contact with deltamethrin may cause a delay/slowness of sensation, inflammation, hypersensitivity, and/or allergy. The action of permethrin takes place in the intercellular spaces and is associated with the possibility of faster decomposition/metabolism, while deltamethrin interacts with receptors, channels, and the cell membrane, which translates into slower decomposition and longer action in the tissue.

Background: Females of Aedes aegypti transmit emerging arboviruses including Zika, dengue, yellow fever, and chikungunya. Control of these adult mosquitoes heavily relies on synthetic insecticides, including pyrethroids. However, insecticide resistance development in populations poses a significant challenge to vector control, particularly from knockdown resistance (kdr) mutations in the voltage-gated sodium channel (VGSC), the target of pyrethroids. This study investigated the field efficacy of Permanone, a pyrethroid-based insecticide, against Ae. aegypti by assessing the impact of three common kdr mutations (V410L, V1016I, F1534C) on mosquito survival under a real operational mosquito control scenario, by quantifying the pesticide delivered in the field., Results: Field cage tests (FCTs) were conducted while conducting a realistic mosquito control application. Female mosquitoes from six operational areas from Harris County, TX, USA were exposed to Permanone delivered with a handheld sprayer. Permanone deposited near the cages was estimated from aluminum boats placed in the field during FCTs using gas chromatography-mass spectrometry (GC-MS). Mortality rates were recorded, and individual mosquitoes were genotyped for kdr mutations. A probit regression model was used to analyze the factors influencing mosquito survivorship. As the distance from the application source route increased, the amount of Permanone deposited decreased, resulting in higher survivorship frequency of Ae. aegypti females with the triple-resistant kdr genotype (LL/II/CC). The L allele at the 410-site significantly contributed to an increased resistance level when co-occurring with other kdr mutations., Conclusion: This study linked the survival probabilities of mosquitoes with different kdr genotypes, and the amount of pesticide they received in the field. Pesticide quantification, control efficacy results and genotyping allowed us to empirically determine the impact of genotypic resistance on vector control in the field. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The black saltmarsh mosquito, Aedes taeniorhynchus, is a prominent nuisance mosquito within St. Johns County, Florida. Due to their characteristically large outbreaks, and the elevated amount of insecticide application correlated with the outbreaks, local populations of Ae. taeniorhynchus are at an increased risk of developing insecticide resistance. This study was established to form a baseline susceptibility of Ae. taeniorhynchus against two technical grade materials, permethrin, and chlorpyrifos. Centers for Disease Control and Prevention bottle bioassays were conducted with technical-grade materials during two outbreaks in the fall of 2023. Results indicated a baseline susceptibility against the materials tested, but most notably, the phenotypic expression of knockdown resistance (kdr) was observed. Results highlight the need for continued monitoring and investigation into the resistance status and resistance level of this common Florida species., (Copyright © 2024 by The American Mosquito Control Association, Inc.)

The cosmopolitan ectoparasite human head louse, Pediculus humanus capitis (De Geer)(Phthiraptera:Pediculidae), affects mostly school-aged children, with infestations reported every year mainly due to louse resistance to pyrethroids. One of the main resistance mechanisms of pyrethroids is the target site insensitivity (kdr), which is caused by single-nucleotide point mutations (SNPs) located in the voltage-sensitive sodium channel gene. In this study, we analyzed individual head lice toxicologically via the description of their susceptibility profile to permethrin and genetically through the genotypification of their kdr alleles as well as nuclear microsatellite loci. Lice were collected from 4 schools in the city of Buenos Aires, Argentina. The resistance ratios varied from 33.3% to 71.4%, with a frequency of the T917I kdr mutation of 87.31% and with 83.6% of the head lice being homozygous resistant to pyrethroids. Microsatellite data indicated that all the louse school populations had genotype proportions that deviated from Hardy-Weinberg expectations, with FIS > 0 reflecting a deficit of heterozygotes. Bottleneck analysis suggested that all louse school populations underwent a recent reduction in population sizes, while 3 of the 4 schools had gene flow values around 1, indicating ongoing gene flow among those schools. Our study suggests that school louse populations in the city of Buenos Aires may form a metapopulation, where each school represents a small population that undergoes extinction and recolonization processes under strong permethrin selection. This is the first multilevel analysis integrating toxicological, kdr-genotyping, and microsatellite data in human louse populations., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Ultra-low volume (ULV) space sprays aerosolize insecticide formulated products (FP) to contact flying mosquitoes, while barrier sprays expose mosquitoes to FP residue on vegetation and other surfaces. Centers for Disease Control and Prevention bottle bioassays used to assess insecticide resistance are based on residual active ingredient (AI) exposure and do not directly relate to FP efficacy. The current pilot study developed a novel compact wind tunnel for mosquito exposure to FP. Caged Aedes albopictus and Culex pipiens/quinquefasciatus were exposed to undiluted Biomist®3 + 15 FP (permethrin AI) or air (control) within the wind tunnel, transferred to new cages, and held in a 28 °C incubator. Separate mosquitoes were exposed to residual permethrin AI (8 μg mL -1 ) in bottle bioassays. Mortality was monitored 15, 30, 60, and 120 min post-exposure., Results: Chi-square tests (P < 0.05) showed significantly higher mortality in Aedes compared to Culex populations for most time points in both bioassay and wind tunnel exposure groups. As expected, mosquitoes exposed to Biomist®3 + 15 showed higher mortality rates than bottle bioassay exposure to permethrin. Two Culex colonies resistant to permethrin in bottle bioassays were susceptible to Biomist®3 + 15 in the wind tunnel., Conclusion: The novel compact wind tunnel developed here may be an alternative to field trials for testing FP efficacy, avoiding factors such as weather, logistical planning, and extended personnel hours. The wind tunnel could allow programs to conveniently test efficacy of multiple FP. Comparisons of different insecticide exposure methods provide practical information to inform operational decisions. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

In addition to killing, mosquito nets treated with permethrin have been claimed to repel mosquitoes, reducing their success in passing through a holed net. We have tested this hypothesis by tracking mosquitoes in a modified World Health Organization tunnel test. In the original assay, mosquitoes are released at one end of the tunnel and have to pass through a holed piece of net to reach the bait at the other end. The mosquitoes are left in the tunnel overnight, while mortality and feeding rates are scored the following morning. Since the original test does not reveal how mosquitoes move within the tunnel, we combined the tunnel with a 3D video camera system. We tracked susceptible and permethrin-resistant Anopheles gambiae s.s. as they moved in the tunnel and interacted with an untreated or a permethrin-treated net (Olyset Net ® ). Surprisingly, while permethrin increased the mortality and reduced blood-feeding rates, twice as many mosquitoes passed through the holes of the permethrin-treated net. The flight trajectories reveal that upon exposure to the permethrin-treated net, both mosquito colonies showed increased 'excitation', thereby augmenting their chance of getting through the holes in the net. The study underlines the complexity of behavioural modes of action of insecticides., (© 2024. The Author(s).)

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.

Furin is a potential target protein associated with numerous diseases; especially closely related to tumors and multiple viral infections including SARS-CoV-2. Most of the existing efficient furin inhibitors adopt a substrate analogous structure, and other types of small molecule inhibitors need to be discovered urgently. In this study, a high-throughput screening combining virtual and physical screening of natural product libraries was performed, coupled with experimental validation and preliminary mechanistic assays at the molecular level, cellular level, and molecular simulation. A novel furin inhibitor, permethrin, which is a derivative from pyrethrin I generated by Pyrethrum cinerariifolium Trev. was identified, and this study confirmed that it binds to a novel allosteric pocket of furin through non-competitive inhibition. It exhibits a very favorable protease-selective inhibition and good cellular activity and specificity. In summary, permethrin shows a new parent nucleus with a new mode of inhibition. It could be used as a highly promising lead compound against furin for targeting related tumors and various resistant viral infections, including SARS-CoV-2.

Long lasting insecticidal nets (LLINs) are a proven tool to reduce malaria transmission, but in Africa efficacy is being reduced by pyrethroid resistance in the major vectors. A previous study that was conducted in Muleba district, Tanzania indicated possible involvement of cytochrome P450 monooxygenases in a pyrethroid resistance in An. gambiae population where pre-exposure to piperonyl butoxide (PBO) followed by permethrin exposure in CDC bottle bioassays led to partial restoration of susceptibility. PBO is a synergist that can block pyrethroid-metabolizing enzymes in a mosquito. Insecticide resistance profiles and underlying mechanisms were investigated in Anopheles gambiae and An. funestus from Muleba during a cluster randomized trial. Diagnostic dose bioassays using permethrin, together with intensity assays, suggest pyrethroid resistance that is both strong and very common, but not extreme. Transcriptomic analysis found multiple P450 genes over expressed including CYP6M2, CYP6Z3, CYP6P3, CYP6P4, CYP6AA1 and CYP9K1 in An. gambiae and CYP6N1, CYP6M7, CYP6M1 and CYP6Z1 in An. funestus. Indeed, very similar suites of P450 enzymes commonly associated with resistant populations elsewhere in Africa were detected as over expressed suggesting a convergence of mechanisms across Sub-Saharan African malaria vectors. The findings give insight into factors that may correlate with pyrethroid PBO LLIN success, broadly supporting model predictions, but revision to guidelines previously issued by the World Health Organization is warranted., Competing Interests: The authors have declared that no competing interests exist.

Aims: A substantial contingent of veterans from the first Gulf War continues to suffer from a number of Gulf War-related illnesses (GWI) affecting the neurological and musculoskeletal systems; the most common symptoms include chronic pain and fatigue. Although animal models have recapitulated several aspects of cognitive impairments in GWI, the pain and fatigue symptoms have not been well documented to allow examination of potential pathogenic mechanisms., Main Methods: We used a mouse model of GWI by exposing mice repeatedly to a combination of Gulf War chemicals (pyridostigmine bromide, permethrin, DEET, and chlorpyrifos) and mild immobilization stress, followed by investigating their pain susceptibilities and fatigue symptoms. To assess whether enhanced antioxidant capacity can counter the effects of GW agents, transgenic mice overexpressing extracellular superoxide dismutase (SOD3OE) were also examined., Key Findings: The mouse model recapitulated several aspects of the human illness, including hyperalgesia, impaired descending inhibition of pain, and increased tonic pain. There is a close association between chronic pain and fatigue in GWI patients. Consistent with this observation, the mouse model showed a significant reduction in physical endurance on the treadmill. Examination of skeletal muscles suggested reduction in mitochondrial functions may have contributed to the fatigue symptoms. Furthermore, the negative impacts of GW agents in pain susceptibilities were largely diminished in SOD3OE mice, suggesting that increased oxidative stress was associated with the emergence of these Gulf War symptoms., Significance: the mouse model will be suitable for delineating specific defects in the pain pathways and mechanisms of fatigue in GWI., (Published by Elsevier Inc.)

Background: The tropical bed bug, Cimex hemipterus, is an important ectoparasite causing various health problems. This species is mainly confined to tropical regions; however, insecticide resistance, global warming, and globalization have changed its distribution map. Molecular information on pyrethroid resistance, which is essential for the development of control programs, is unknown for C. hemipterus in expanded areas. The present study was designed to determine the permethrin resistance status, characterize the pyrethroid receptor sites in voltage-gated sodium channel (vgsc) gene, and identify the resistance-related mutations in the populations of tropical bed bug in Iran., Methods: Live bed bugs were collected, and adults of C. hemipterus were selected for bioassay and molecular surveys. Bioassay was performed by tarsal contact with permethrin 0.75% in mixed-sex of samples. Conventional and quantitative TaqMan and SYBR Green real-time PCR assays were conducted to characterize the vgsc gene and genotypes of studied populations., Results: In the bioassay tests, the mortality rates were in the range of 30.7-38.7% and 56.2-77.4% in 24 and 48 h, respectively. The knockdown rates of studied populations were in the range of 32.2-46.6% and 61.5-83.8% in the first and second days, respectively. The KT 50 and KT 90 values in the Cimex lectularius Kh1 strain were presented as 5.39 and 15.55 h, respectively. These values in the selected populations of C. hemipterus varied from 27.9 to 29.5 and from 82.8 to 104.4 h, respectively. Knockdown time ratios (KR 50 and KR 90 ) in these populations varied from 5.17 to 6.17-fold compared with those of the C. lectularius Kh1 strain. Fragments of vgsc gene with 355 bp and 812 bp were amplified. Analysis of sequences revealed the A468T substitution, kdr-associated D953G, and super-kdr M918I and L1014F mutations in all populations., Conclusions: The specific/sensitive, safe, and rapid diagnostic assays developed in this study are recommended for detection of kdr/super-kdr mutations and frequency of mutant alleles. The presence of super-kdr mutations and high resistance to permethrin in all the populations necessitate the reconsideration of control approaches against C. hemipterus., (© 2021. The Author(s).)

Objectives: To assess the effectiveness of nanopermethrin as a potential new formulation for pest and vector control., Methods: Permethrin nanoparticles were prepared by the ionic gelation method and its structure and the formulations were designed using Box-Behnken statistical technique. The effect of independent variables (Chitosan/Permethrin ratio, tripolyphosphate quantity, sonication time) on the properties of nanoparticles was investigated to determine the optimal formulation., Results: The size of the nanoparticles ranged from 135.27 ± 5.88 to 539.5 ± 24.01 nm and the insecticide entrapment efficiency per cent (EE%) ranged from 7.72 ± 1.36 to 63.59 ± 3.17%. Anopheles stephensi larvae were then bioassayed with the nanopermethrin and compared with the results of the bioassay with the mother molecule of permethrin using a standard WHO-recommended mosquito larval bioassay kit. LC 50 with permethrin and nanopermethrin on larvae of An. stephensi were 0.125 and 0.026 ppm showing a 4.8 times difference. The LC 50 for permethrin and nanopermethrin on Culex pipiens were 0.003 and 0.00032 ppm, respectively, showing a 9.4-fold difference., Conclusion: Nanopermethrin is much more potent than its mother molecule against larvae of An. stephensi and Cx. pipiens., (© 2021 John Wiley & Sons Ltd.)

Background: The selection and the spread of insecticide resistance in malaria vectors to the main classes of insecticides used in vector control tools are a major and ongoing challenge to malaria vector control programmes. This study aimed to determine the intensity of vector resistance to insecticides in three regions of Benin with different agro-ecological characteristics., Methods: Larvae of Anopheles gambiae sensu lato (s.l.) were collected from September to November 2017 in different larval sites in three northern Benin communes: Parakou, Kandi and Malanville. Two to five-day-old, non-blood-fed, female mosquitoes were exposed to papers impregnated with deltamethrin, permethrin and bendiocarb at dosages of 1 × the diagnostic dose, 5 × and 10 × to determine the intensity of resistance in these vectors. Molecular frequencies of the kdr L1014F and ace-1R G119S insecticide resistance mutations and levels of detoxification enzymes were determined for mosquitoes sampled at each study site., Results: Resistance to pyrethroids (permethrin and deltamethrin) was recorded in all three communes with mortality rates below 60% using the diagnostic dose (1x). The results obtained after exposure of An. gambiae to permethrin 10 × were 99% in Kandi, 98% in Malanville and 99% in Parakou. With deltamethrin 10x, mortality rates were 100% in Kandi, 96% in Malanville and 73% in Parakou. For the diagnostic dose of bendiocarb, suspected resistance was recorded in the communes of Malanville (97%) and Kandi (94%) while sensitivity was observed in Parakou (98%).Using the 10 × dose, mortality was 98% in Kandi, 100% in Malanville and 99% in Parakou. The frequencies of the kdr L1014F allele varied between 59 and 83% depending on the sites and species of the An. gambiae complex, while the frequency of the ace-1R G119S gene varied between 0 and 5%. Biochemical tests showed high levels of oxidase and esterase activity compared to the susceptible colony strain of An. gambiae sensu stricto (Kisumu strain)., Conclusion: Anopheles gambiae showed a generalized loss of susceptibility to permethrin and deltamethrin but also showed moderate to high intensity of resistance in different regions of Benin. This high intensity of resistance is a potential threat to the effectiveness of vector control., (© 2021. The Author(s).)

Pesticide resistance in medically significant disease vectors can negatively impact the efficacy of control efforts. Resistance research on ticks has focused primarily on species of veterinary significance that experience relatively high degrees of control pressure. Resistance in tick vectors of medical significance has received little attention, in part because area-wide pesticide applications are not used to control these generalist tick species. One of the few effective methods currently used for area-wide control of medically important ticks, including Ixodes scapularis Say (Acari: Ixodidae), is deployment of 4-poster devices. Deer self-apply a topical acaricide (permethrin) while feeding on corn from the devices. A 4-poster program using permethrin has been deployed on Shelter Island, NY to control I. scapularis populations since 2008. We collected engorged female ticks from deer in this management area and a location in the Mid-Hudson River Valley, NY without area-wide tick control. Larvae were reared from egg masses and their susceptibility to permethrin was tested. Larvae originating from a long-term laboratory colony were used as a susceptible baseline for comparison. Compared against the laboratory colony, resistance ratios at LC-50 for Shelter Island and Hudson Valley I. scapularis were 1.87 and 1.51, respectively. The susceptibilities of the field populations to permethrin were significantly lower than that of the colony ticks. We provide the first data using the larval packet test to establish baseline susceptibility for I. scapularis to permethrin along with information relevant to understanding resistance emergence in tick populations under sustained control pressure from 4-poster devices., (Published by Oxford University Press on behalf of Entomological Society of America 2021.)

Insecticide resistance has emerged as a persistent threat to the fight against vector-borne diseases. We compared the gut microbiota of permethrin-selected (PS) strain of Aedes aegypti relative to the parent (KW) strain from Key West, Florida. Bacterial richness but not diversity was significantly higher in PS strain compared to KW strain. The two mosquito strains also differed in their gut microbial composition. Cutibacterium spp., Corynebacterium spp., Citricoccus spp., Leucobacter spp., Acinetobacter spp., Dietzia spp., and Anaerococcus spp. were more abundant in PS strain than in KW strain. In contrast, Sphingomonas spp., Aquabacterium spp., Methylobacterium spp., Flavobacterium spp., Lactobacillus spp., unclassified Burkholderiaceae and unclassified Nostocaceae were more abundant in KW strain compared to PS strain. PS strain was enriched with propionate metabolizers, selenate reducers, and xylan, chitin, and chlorophenol degraders while KW strain was enriched with sulfur oxidizers, sulfur metabolizers, sulfate reducers and naphthalene and aromatic hydrocarbons degraders. These findings demonstrate an association between the gut microbiota and insecticide resistance in an important vector species and sets the foundation for future studies to investigate the contribution of gut microbiota to evolution of insecticide resistance in disease vectors., (© 2021. The Author(s).)

Pyrethroids are one of the few classes of insecticides available to control Aedes aegypti, the major vector of dengue, chikungunya, and Zika viruses. Unfortunately, evolving mechanisms of pyrethroid resistance in mosquito populations threaten our ability to control disease outbreaks. Two common pyrethroid resistance mechanisms occur in Ae. aegypti: 1) knockdown resistance, which involves amino acid substitutions at the pyrethroid target site-the voltage-gated sodium channel (VGSC)-and 2) enhanced metabolism by detoxification enzymes. When a heterogeneous population of mosquitoes is exposed to pyrethroids, different responses occur. During exposure, a proportion of mosquitoes exhibit immediate knockdown, whereas others are not knocked-down and are designated knockdown resistant (kdr). When these individuals are removed from the source of insecticide, the knocked-down mosquitoes can either remain in this status and lead to dead or recover within a few hours. The proportion of these phenotypic responses is dependent on the pyrethroid concentration and the genetic background of the population tested. In this study, we sequenced and performed pairwise genome comparisons between kdr, recovered, and dead phenotypes in a pyrethroid-resistant colony from Tapachula, Mexico. We identified single-nucleotide polymorphisms (SNPs) associated with each phenotype and identified genes that are likely associated with the mechanisms of pyrethroid resistance, including detoxification, the cuticle, and insecticide target sites. We identified high association between kdr and mutations at VGSC and moderate association with additional insecticide target site, detoxification, and cuticle protein coding genes. Recovery was associated with cuticle proteins, the voltage-dependent calcium channel, and a different group of detoxification genes. We provide a list of detoxification genes under directional selection in this field-resistant population. Their functional roles in pyrethroid metabolism and their potential uses as genomic markers of resistance require validation., Competing Interests: The authors have declared that no competing interests exist. The findings and conclusions in this paper are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention or the National Institutes of Health. Author Americo Rodriguez was unable to confirm their authorship contributions. On their behalf, the corresponding author has reported their contributions to the best of their knowledge.

The effects of permethrin (PRM) and deltamethrin (DLM) on acoustic or light prepulse inhibition of the acoustic startle response (ASR) and tactile startle response (TSR) were studied in adult male Sprague Dawley rats. Preliminary studies were conducted to optimize the parameters of light and acoustic prepulse inhibition of ASR and TSR. Once these parameters were set, a new group of rats was administered PRM (0 or 90 mg/kg) or DLM (0 or 25 mg/kg) by gavage in 5 mL/kg corn oil. ASR and TSR were assessed using acoustic or light prepulses 6, 8, and 12 h after PRM and 2, 4, and 6 h after DLM exposure. PRM increased ASR 6 h post-treatment with no interaction with acoustic prepulse levels and with no effect on TSR. When light was used as the prepulse, PRM increased ASR and TSR at 6 h with no interaction with prepulse levels. DLM decreased ASR and TSR on trials without prepulses but not on trials with acoustic prepulses. DLM also decreased ASR when light prepulses were present 4 h post-treatment. A final experiment assessed whether the house light in the test cabinet affected ASR and TSR after PRM or DLM exposure. Rats had increased ASR and TSR when house lights were on compared with when they were off, but lighting did not differentially interact with PRM or DLM. Light and acoustic prepulses of ASR and TSR have different effects depending on the test agent and the test parameters.

The brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.) Latreille (Acari: Ixodidae), is a peridomestic, cosmopolitan parasite of dogs known to vector numerous pathogens of veterinary and medical importance. Recent phylogenetic analyses separate this tick into temperate and tropical lineages. Populations of Rh. sanguineus s.l. have been reported to exhibit sodium channel target site insensitivity to permethrin and etofenprox, which is likely due to the prolonged use of pyrethroids against many pests in and around the home. In this study, populations collected in the Caribbean, Africa, Asia, Europe and North America, were tested to identify the distribution of a known resistance mechanism, pathogen-vector interactions and phylogeny in relation to latitude. Using molecular assays, populations from 29 distinct locations were simultaneously geographically typed and screened for bacterial infection by Rickettsia, Ehrlichia, Babesia and Hepatozoon species, and for the presence of a sodium channel single nucleotide polymorphism known to confer permethrin resistance. Implications of these results on Rh. sanguineus s.l. management in association with geographical distribution will be discussed., (© 2020 The Royal Entomological Society.)

Insecticides, especially pyrethroids, are the most important in the insect pest control and preventing insect vector-borne human diseases. However, insect pests, including mosquitoes, have developed resistance in the insecticides that used against them. Cytochrome P450s are associated with insecticide resistance through overexpression and detoxification mechanisms in insect species. In this study, we utilized a powerful tool, the RNAi technique, to determine the roles of key P450 genes overexpressed in permethrin resistant mosquitoes that confer insecticide resistance to unravel the molecular basis of resistance mechanisms in the mosquito Culex quinquefasciatus. The results showed that knockdown of 8 key P450 genes using RNAi techniques significantly decreased resistance to permethrin in resistant mosquitoes. In silico modeling and docking analysis further revealed the potential metabolic function of overexpressed P450 genes in the development of insecticide resistance in mosquitoes. These findings not only highlighted the functional importance of these P450 genes in insecticide resistance, but also revealed that overexpression of multiple P450 genes was responsible for the high levels of insecticide resistance in a mosquito population of Culex quinquefasciatus.

Background: Insecticide resistance is threatening the effectiveness of efforts to control malaria vectors in Benin. This study explores the levels and mechanisms of insecticide resistance in An. gambiae s.l. to pyrethroids., Methods: Larvae were collected from August 2017 to July 2018 in five communes in southern Benin (Adjohoun, Allada, Bohicon, Cotonou, and Porto-Novo) representing diverse ecological regions, and were reared in Benin's insectary. Two- to five-day-old female mosquitoes from each district were exposed to multiple doses of deltamethrin and permethrin (1×, 2×, 5×, and 10×) using the WHO insecticide resistance intensity bioassay. The effect of pre-exposure to the synergist, piperonyl butoxide (PBO), was also tested at different pyrethroid doses. Molecular allele frequencies of kdr (1014F) and ace-1R (119S) insecticide resistance mutations and levels of detoxification enzymes were determined for mosquitoes sampled from each study area., Results: An. gambiae s.l. were resistant to pyrethroid-only exposure up to 10× the diagnostic doses in all the study sites for both deltamethrin and permethrin. Mortality was significantly higher in An. gambiae s.l. pre-exposed to PBO followed by exposure to deltamethrin or permethrin compared to mosquitoes exposed to deltamethrin or permethrin only (p < 0.001). The difference in mortality between deltamethrin only and PBO plus deltamethrin was the smallest in Cotonou (16-64%) and the greatest in Bohicon (12-93%). The mortality difference between permethrin only and PBO plus permethrin was the smallest in Cotonou (44-75%) and the greatest in Bohicon (22-72%). In all the study sites, the kdr resistance allele (1014F) frequency was high (75-100%), while the ace-1 resistance allele (G119S) frequency was low (0-3%). Analysis of the metabolic enzymatic activity of An. gambiae s.l. showed overexpression of nonspecific esterases and glutathione S-transferases (GST) in all study sites. In contrast to the PBO results, oxidase expression was low and was similar to the susceptible An. gambiae s.s. Kisumu strain in all sites., Conclusion: There is high-intensity resistance to pyrethroids in southern Benin. However, pre-exposure to PBO significantly increased susceptibility to the pyrethroids in the different An. gambiae s.l. populations sampled. The use of PBO insecticide-treated bed nets may help maintain the gains in An. gambiae (s.l.) control in southern Benin.

There are major public health concerns regarding the spread of mosquito-borne diseases such as dengue, Zika, and chikungunya, which are mainly controlled by using insecticides against the vectors, Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse). Pyrethroids are the primary class of insecticides used for vector control, due to their rapid knockdown effect and low toxicity to vertebrates. Unfortunately, continued use of pyrethroids has led to widespread insecticide resistance in Ae. aegypti; however, we lack information for Ae. albopictus-a sympatric species in Chiapas since 2002. In this study, we evaluated the permethrin resistance status of Ae. albopictus collected from Mexico and Texas. We also selected for permethrin resistance in the laboratory and investigated the potential mechanisms conferring resistance in this species. Knockdown resistance mutations, specifically F1534C, in the voltage-gated sodium channel gene, and increased activity of detoxifying enzymes were evaluated. Low levels of permethrin resistance (<2.4-fold) were observed in our field populations of Ae. albopictus and the F1534C mutation was not detected in any of the sites. Low levels of resistance were also observed in the artificially selected strain. There was significantly higher cytochrome P450 activity in our permethrin-selected and nonselected strains from Mexico compared to the control strain. Our results suggest the Ae. albopictus sampled from 2016 are mostly susceptible to pyrethroids. These results contrast with the high levels of permethrin resistance (>58-fold) found in Ae. aegypti from the same sites in Mexico. This research indicates the importance of continued monitoring of Ae. albopictus populations to prevent resistance from developing in the future., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: The risk of tick-borne diseases is decreased by increasing awareness and knowledge through prevention education. The aim of the present study was to evaluate the effect of long-lasting permethrin impregnated (LLPI) socks for tick bites., Methods: A randomized open label study was conducted to determine the effectiveness of LLPI socks for prevention of tick bites among 367 adults living in a rural area. Participants completed questionnaires at the start of follow-up (July 2014) and at the end of follow-up (December 2014), and tick bites were reported., Results: A total of 332 subjects completed the follow-up survey. The tick bite rate of the two groups was not significantly different (3.6% vs. 3.1%). But the tick bite rate of lower extremities of subjects wearing LLPI socks was significantly lower compared to that of subjects wearing general socks., Conclusion: The tick bite rate was not different between the two groups, but the tick bite rate of lower extremities of LLPI was significantly lower than general groups. Further study is needed to investigate the effect of LLPI clothes with larger populations., Competing Interests: The authors have no potential conflicts of interest to disclose., (© 2021 The Korean Academy of Medical Sciences.)

Background: Vector control for malaria prevention relies most often on the use of insecticide-treated bed net (ITNs) and indoor residual spraying. Little is known about the longevity of long-lasting insecticidal nets (LLINs) in the Americas. The physical integrity and insecticide retention of LLINs over time were monitored after a bed net distribution campaign to assess community practices around LLIN care and use in Waspam, northeastern Nicaragua., Methods: At least 30 nets were collected at 6, 12, 24, and 36 months post distribution. Physical integrity was measured by counting holes and classifying nets into categories (good, damaged, and too torn) depending on a proportionate hole index (pHI). Insecticide bioefficacy was assessed using cone bioassays, and insecticide content measured using a cyanopyrethroid field test (CFT)., Results: At 6 months, 87.3 % of LLINs were in good physical condition, while by 36 months this decreased to 20.6 %, with 38.2 % considered 'too torn.' The median pHI increased from 7 at the 6-month time point to 480.5 by 36 months. After 36 months of use, median mortality in cone bioassays was 2 % (range: 0-6 %) compared to 16 % (range: 2-70 %) at 6 months. There was a decrease in the level of deltamethrin detected on the surface of the LLINs with 100 % of tested LLINs tested at 12 months and 24 months crossing the threshold for being considered a failed net by CFT., Conclusions: This first comprehensive analysis of LLIN durability in Central America revealed rapid loss of chemical bioefficacy and progressive physical damage over a 36-month period. Use of these findings to guide future LLIN interventions in malaria elimination settings in Nicaragua, and potentially elsewhere in the Americas, could help optimize the successful implementation of vector control strategies.

Background: Insecticide resistance poses a growing challenge to malaria vector control in Kenya and around the world. Following evidence of associations between the mosquito microbiota and insecticide resistance, the microbiota of Anopheles gambiae sensu stricto (s.s.) from Tulukuyi village, Bungoma, Kenya, with differing permethrin resistance profiles were comparatively characterized., Methods: Using the CDC bottle bioassay, 133 2-3 day-old, virgin, non-blood fed female F 1 progeny of field-caught An. gambiae s.s. were exposed to five times (107.5 µg/ml) the discriminating dose of permethrin. Post bioassay, 50 resistant and 50 susceptible mosquitoes were subsequently screened for kdr East and West mutations, and individually processed for microbial analysis using high throughput sequencing targeting the universal bacterial and archaeal 16S rRNA gene., Results: 47 % of the samples tested (n = 133) were resistant, and of the 100 selected for further processing, 99 % were positive for kdr East and 1 % for kdr West. Overall, 84 bacterial taxa were detected across all mosquito samples, with 36 of these shared between resistant and susceptible mosquitoes. A total of 20 bacterial taxa were unique to the resistant mosquitoes and 28 were unique to the susceptible mosquitoes. There were significant differences in bacterial composition between resistant and susceptible individuals (PERMANOVA, pseudo-F = 2.33, P = 0.001), with presence of Sphingobacterium, Lysinibacillus and Streptococcus (all known pyrethroid-degrading taxa), and the radiotolerant Rubrobacter, being significantly associated with resistant mosquitoes. On the other hand, the presence of Myxococcus, was significantly associated with susceptible mosquitoes., Conclusions: This is the first report of distinct microbiota in An. gambiae s.s. associated with intense pyrethroid resistance. The findings highlight differentially abundant bacterial taxa between resistant and susceptible mosquitoes, and further suggest a microbe-mediated mechanism of insecticide resistance in mosquitoes. These results also indicate fixation of the kdr East mutation in this mosquito population, precluding further analysis of its associations with the mosquito microbiota, but presenting the hypothesis that any microbe-mediated mechanism of insecticide resistance would be likely of a metabolic nature. Overall, this study lays initial groundwork for understanding microbe-mediated mechanisms of insecticide resistance in African mosquito vectors of malaria, and potentially identifying novel microbial markers of insecticide resistance that could supplement existing vector surveillance tools.

Human head lice are blood-sucking insects causing an infestation in humans called pediculosis capitis. The infestation is more prevalent in the school-aged population. Scalp itching, a common presenting symptom, results in scratching and sleep disturbance. The condition can lead to social stigmatization which can lead to loss of self-esteem. Currently, the mainstay of treatment for pediculosis is chemical insecticides such as permethrin. The extended use of permethrin worldwide leads to growing pediculicide resistance. The aim of this study is to demonstrate the presence of the knockdown resistance (kdr) mutation in head lice populations from six different localities of Thailand. A total of 260 head lice samples in this study were collected from 15 provinces in the 6 regions of Thailand. Polymerase chain reaction (PCR) was used to amplify the α subunit of voltage-sensitive sodium channel (VSSC) gene, kdr mutation (C→T substitution). Restriction fragment length polymorphism (RFLP) patterns and sequencing were used to identify the kdr T917I mutation and demonstrated three genotypic forms including homozygous susceptible (SS), heterozygous genotype (RS), and homozygous resistant (RR). Of 260 samples from this study, 156 (60.00%) were SS, 58 (22.31%) were RS, and 46 (17.69%) were RR. The overall frequency of the kdr T917I mutation was 0.31. Genotypes frequencies determination using the exact test of Hardy-Weinberg equilibrium found that northern, central, northeastern, southern, and western region of Thailand differed from expectation. The five aforementioned localities had positive inbreeding coefficient value (Fis > 0) which indicated an excess of homozygotes. The nucleotide and amino acid sequences of RS and RR showed T917I and L920F point mutations. In conclusion, this is the first study detecting permethrin resistance among human head lice from Thailand. PCR-RFLP is an easy technique to demonstrate the kdr mutation in head louse. The data obtained from this study would increase awareness of increasing of the kdr mutation in head louse in Thailand., Competing Interests: The authors have declared that no competing interests exist.

Culex quinquefasciatus is one of the most important mosquito vectors of arboviruses. Currently, the fastest approach to control disease transmission is the application of synthetic adulticide insecticides. However, in highly populated urban centers the development of insecticide resistance in mosquito populations could impair insecticide efficacy and therefore, disease control. To assess the effect of resistance on vector control, females of Cx. quinquefasciatus collected from six mosquito control operational areas in Harris County, Texas, were treated in field cage tests at three different distances with the pyrethroid Permanone® 31-66 applied at the operational rate. Females were analyzed by sequencing and/or diagnostic PCR using de novo designed primers for detecting the kdr-like mutation in the voltage-gated sodium channel (L982F; TTA to TTT) (house fly kdr canonical mutation L1014F). Females from the Cx. quinquefasciatus susceptible Sebring strain and those from the six operational areas placed at 30.4 m from the treatment source were killed in the tests, while 14% of field-collected mosquitoes survived at 60.8 m, and 35% at 91.2 m from the source. The diagnostic PCR had a with 97.5% accuracy to detect the kdr-like mutation. Pyrethroid resistant mosquitoes carrying the L982F mutation were broadly distributed in Harris County at high frequency. Among mosquitoes analyzed (n = 1,028), the kdr-kdr genotype was prevalent (81.2%), the kdr-s genotype was 18%, and s-s mosquitoes were less than 1% (n = 8). A logistic regression model estimated an equal probability of survival for the genotypes kdr-kdr and kdr-s in all areas analyzed. Altogether, our results point to a high-risk situation for the pyrethroid-based arboviral disease control in Harris County., Competing Interests: The authors have declared that no competing interests exist.

Background: The correct treatment and management of scabies is expensive and time-consuming, and may have a negative impact on patients and their families., Aim: To investigate the effects of permethrin 5% cream on scabies mites, and explore mite survival times outside the human body., Methods: We performed a nonrandomized controlled study. In total, 20 petri dishes were coated with permethrin 5% cream (treatment group) and 20 plain petri dishes (control group) each had one scabies mite placed in them, and were then observed at baseline and 3, 4, 5, 6, 7, 8 and 12 h from baseline. In the second part of our study, 30 scabies mites from infested patients were investigated in an observational experiment in 30 plain petri dishes at days 0, 3 and 4., Results: Our data showed that 65% of scabies mites survived after 8 h in the treatment group compared with 75% of mites in the control group. After 12 h, 25% of mites in the treatment group and 60% in the control group were still alive. Data from the observational survival study showed that one mite was alive on day 3, but all mites were dead by day 4., Conclusions: This study showed no significant effects of mite survival times with 5% topical permethrin after 8 h, while its efficacy was stronger and significant after 12 h. We recommend the isolation of all mite-infested items for at least 4 days., (© 2020 British Association of Dermatologists.)

Antibiotic-resistant strains of bacteria such as methicillin-resistant Staphylococcus aureus are a threat to human health, and effective treatment options against them are needed. This study aimed to determine whether the insecticide permethrin was capable of inhibiting the growth of S. aureus or if some other component of a permethrin cream was responsible for a decrease in scabies associated bacterial infection previously observed. Ten S. aureus strains were grown in the presence of permethrin and formaldehyde both alone and in combination with percent inhibition determined by viable counts. Also, a time-kill assay was conducted on S. aureus exposed to the same conditions. Finally, the morphology of S. aureus grown in the presence of permethrin was examined by scanning electron microscopy. Bacterial inhibition by permethrin ranged from 0% to 41% whereas inhibition by formaldehyde was 100%. The time-kill curves of permethrin exposed cells were very similar to the positive growth control while the formaldehyde and combination exposure showed complete inhibition even at the 0-hr time point. The scanning electron micrographs of permethrin grown S. aureus showed healthy cocci cells with no sign of cell damage. Our results show that permethrin is not capable of inhibiting the growth of bacteria enough for it to be termed bactericidal. Formaldehyde is a known antiseptic and therefore was responsible for the antibacterial effect observed after the use of permethrin cream., (© 2020 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

The use of insecticides has been a central approach to control disease-transmitting mosquitoes for the last century. The high prevalence of pyrethroid use as public health insecticides has resulted in the evolution of pyrethroid resistance in many populations of Aedes aegypti (Linnaeus) (Diptera: Culicidae), throughout its global distribution range. Insecticide resistance is often correlated with an associated fitness cost. In this project, we studied the phenotypes of hybrid mosquitoes derived from crossing a pyrethroid-resistant strain of Ae. aegypti (Puerto Rico [PR]) with a more susceptible one (Rockefeller [ROCK]). We first sequenced and compared the para gene of both original strains. We then crossed males from one strain with females of the other, creating two hybrids (Puertofeller, Rockorico). We used a Y-tube choice assay to measure the attraction of these strains towards a human host. We then compared the levels of pyrethroid resistance in the different strains. We found three known resistance mutations in the para gene sequence of the PR strain. In our attraction assays, PR females showed lower attraction to humans, than the ROCK females. Both hybrid strains showed strong attraction to a human host. In the insecticide resistance bottle assays, both hybrid strains showed marginal increases in resistance to permethrin compared to the more susceptible ROCK strain. These results suggest that hybrids of sensitive and permethrin-resistant mosquitoes have an incremental advantage compared to more susceptible mosquitoes when challenged with permethrin. This explains the rapid spread of permethrin resistance that was observed many times in the field., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.)

The cattle tick, Rhipicephalus annulatus (Say) is a vector of bovine babesiosis and responsible for direct and indirect losses to cattle producing areas located in temperate and subtropical dry regions. Resistance against pyrethroids has been reported for this species in Asia and Africa, but never before in North America. An outbreak strain, Rio Lado, collected close to the border between Mexico and the United States, in Maverick County, Texas, showed low level of resistance to permethrin, a pyrethroid pesticide. We used genomic material from different strains of cattle ticks collected within the Permanent Quarantine Zone (Rio Lado, Vega and Klein Grass strains) to partially characterize the coding gene of the voltage-gated sodium channel (Na-channel), target-site of pyrethroid pesticides, and search for putative mutations associated with resistance using quantitative PCR high resolution melt (HRM) analysis. The two amplified fragments, corresponding to domains II and III of the Na-channel, were 100 % identical to its ortholog in Rhipicephalus microplus (Canestrini). No nucleotide polymorphisms in the Na-channel gene were observed in the pyrethroid-resistant Rio Lado strain, when compared to the susceptible strains Klein Grass and Vega. This study reports the first case of pyrethroid resistance in R. annulatus collected in the United States. Also, we provide new genomic data for this species of tick that allows for the development of a new method to screen for mutations associated with pyrethroid resistance., Competing Interests: Declaration of Competing Interest None to declare., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Pyrethroid pesticides are extensively used to manage animal and human disease vectors including the southern cattle tick Rhipicephalus microplus (Canestrini). The indiscriminate and incorrect use of pyrethroids has led to the almost ubiquitous development of resistance to this pesticide class for this tick species. Voltage-gated sodium channels (Na-channels) are the primary target-site of pyrethroids and several studies on the involvement of mutations in the coding gene among pyrethroid-resistant R. microplus populations from different parts of the world have shown that there are various single nucleotide polymorphisms (SNPs) that are associated with resistance to pyrethroids. Identification of the exact location of the mutations in the protein coding regions of the targeted gene facilitates the design of various molecular tools for genotyping the resistant populations and thus promotes the rapid detection of resistance. This review aims to provide an update on the identification of pyrethroid resistance-associated Na-channel mutations from R. microplus.The database of diverse mutations from different regions of the world helped us to develop the molecular markers for resistance monitoring in a rapid and efficient manner. Their role and the development of different forms of molecular tools for genotyping ticks for mutations in the Na-channel gene are also discussed. In this review, the word mutation is used interchangeably with SNP., (Copyright © 2020. Published by Elsevier GmbH.)

Head lice infestation caused by Pediculus humanus capitis De Geer, 1767 is one of the most common public health problems. The relationship between humans and head lice dates back millions of years ago that differentiated into different phylogenetic clades. Treatment of head lice infestation usually based on insecticide-based products, which promotes the resistance in the head lice populations. In the present study, we aimed to screen the presence of permethrin resistance among collected P. h. capitis specimens in Turkey. Three mutation sites (T917I, L920F, and M815I) were screened using real-time PCR and resistance was identified by melt analysis. Of the studied specimens, resistance allele frequency (RAF) was found 0.98 for T917I, 0.99 for L920F, and 1.00 for M815I. The phylogenetic study revealed that Clade A and Clade B are present and overlap in Turkey. The present study is first to screen the resistance among Turkish head lice specimens. To not stimulate the pyrethroids resistance in head lice populations, early detection of resistance is crucial and will help the health professionals to choose suitable formula in the treatment. We suggest that the resistance status needs to be screened in randomly selected populations before any treatment application is given., Competing Interests: Declaration of Competing Interest On behalf of all authors, the corresponding author states that there is no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Mosquitoes and other vectors are often exposed to sublethal doses of insecticides. Larvae can be exposed to the run-off of agricultural use, and adults can be irritated by insecticides used against them and move away before they have picked up a lethal dose. This sublethal exposure may affect the success of control of insect-borne diseases, for it may affect the competence of insects to transmit parasites, in particular if the insects are undernourished., Methods: We assessed how exposure of larvae and adults to a sublethal dose of permethrin (a pyrethroid) and how larval competition for food affect several aspects of the vector competence of the mosquito Anopheles gambiae for the malaria parasite Plasmodium berghei. We infected mosquitoes with P. berghei and measured the longevity and the prevalence and intensity of infection to test for an effect of our treatments., Results: Our general result was that the exposure to the insecticide helped mosquitoes deal with infection by malaria. Exposure of either larvae or adults decreased the likelihood that mosquitoes were infected by about 20%, but did not effect the parasite load. Exposure also increased the lifespan of infected mosquitoes, but only if they had been reared in competition. Larval competition had no effect on the prevalence of infection, but increased parasite load. These effects may be a consequence of the machinery governing oxidative stress, which underlies the responses of mosquitoes to insecticides, to food stress and to parasites., Conclusions: We conclude that insecticide residues are likely to affect the ability of mosquitoes to carry and transmit pathogens such as malaria, irrespective of the stage at which they are exposed to the insecticide. Our results stress the need for further studies to consider sublethal doses in the context of vector ecology and vector-borne disease epidemiology.

The insecticides we use for agriculture and for vector control often arrive in water bodies, where mosquito larvae may be exposed to them. Not only will they then likely affect the development of the larvae, but their effects may carry over to the adults, potentially affecting their capacity at transmitting infectious diseases. Such an impact may be expected to be more severe when mosquitoes are undernourished. In this study, we investigated whether exposing larvae of the mosquito Anopheles gambiae to a sub-lethal dose of permethrin (a pyrethroid) and forcing them to compete for food would affect the immune response of the adults. We found that a low dose of permethrin increased the degree to which individually reared larvae melanised a negatively charged Sephadex bead and slowed the replication of injected Escherichia coli. However, if mosquitoes had been reared in groups of three (and thus had been forced to compete for food) permethrin had less impact on the efficacy of the immune responses. Our results show how larval stressors can affect the immune response of adults, and that the outcome of exposure to insecticides strongly depends on environmental conditions.

In addition to the direct effect of insecticide-treated nets (ITNs), there has been evidence for spatial indirect effects. Spatial analyses in cluster randomized trials (CRTs) are rare, but a large-scale CRT from 1993 was one of the first to conduct a spatial analysis of ITNs in CRTs. We revisit these data by applying a broader range of contemporary spatial methods to further explore spatial spillover. We conducted three analyses: 1) exploratory spatial analysis, considering spatial patterns and spillover in the data; 2) spatial modeling, estimating the intervention effect considering spatial effects; and 3) analysis of distance-based spillover and interaction with the intervention, characterizing the functional distance over which the spillover effect was present. There were consistent indications of spatial patterns from the exploratory analysis. Bed nets were associated with a 17% reduction in all-cause mortality for children aged 6-59 months, and the intervention estimate remained robust when allowing for the spatial structure of the data. There was strong evidence of a spatial spillover effect: for every additional 100 m that a control household was from an intervention household (and vice versa), the standardized mortality ratio (SMR) increased by 1.7% (SMR 1.017, 95% credible interval 1.006-1.026). Despite evidence of a spatial spillover effect, the conclusions of the trial remain unaffected by spatial model specifications. Use of ITNs was clearly beneficial for individuals, and there was compelling evidence that they provide an indirect benefit to individuals living nearby. This article demonstrates the extra utility that spatial methods can provide when analyzing a CRT.

Permethrin (PM) is a synthetic pyrethroid insecticide widely used as domestic repellent. Damage effects to nontarget organisms have been reported, particularly in the early stages of development. Studies indicate redox unbalance as secondary PM effect. Therefore, our goal was to investigate the acute PM effects on larval zebrafish. Larvae (6 days postfertilization) were exposed to PM (25-600 μ g/L) during 24 hours, and 50% lethal concentration was estimated. For subsequent assays, the sublethal PM concentrations of 25 and 50 μ g/L were used. PM increased anxiety-like behaviors according to the Novel Tank and Light-Dark tests. At the molecular level, PM induced increased ROS, which may be related to the increased lipid peroxidation, DNA damage, and apoptosis detected in PM-exposed organisms. In parallel, upregulation of the antioxidant system was detected after PM exposure, with increased superoxide dismutase, glutathione S-transferase and glutathione reductase activities, and thiol levels. The increased of Nrf2 target genes and the activation of an electrophile response element-driven reporter Tg( EPRE :LUC-EGFP) suggest that the Nrf2 pathway can mediate a fast response to PM, leading to antioxidant amplification. By using high-resolution respirometry, we found that exposure to PM decreased the oxygen consumption in all respiratory stages, disrupting the oxidative phosphorylation and inhibiting the electron transfer system, leading to decrease in bioenergetics capacity. In addition, PM led to increases of residual oxygen consumption and changes in substrate control ratio. Glucose metabolism seems to be affected by PM, with increased lactate dehydrogenase and decreased citrate synthase activities. Taken together, our results demonstrated the adverse effects of acute sublethal PM concentrations during larval development in zebrafish, causing apparent mitochondrial dysfunction, indicating a potential mechanism to redox unbalance and oxidative stress, which may be linked to the detected cell death and alterations in normal behavior patterns caused by acute PM exposure., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2019 Mauro Eugênio Medina Nunes et al.)

Background: Permethrin (a pyrethroid) has been recommended for the management of a number of insect pests, including Musca domestica. Recently, resistance to permethrin has been reported from the Punjab province, Pakistan, which necessitates the need to manage the resistance problem. For this reason, a field strain of M. domestica was selected with permethrin for ten consecutive generations (G1-G10) to determine the risk assessment, the rapidity with which permethrin resistance can develop, and the presence or absence of the cross-resistance phenomenon., Results: The results revealed that permethrin resistance increased very quickly in response to selection experiments. The resistance ratio (RR) value increased by 16.04- to 605.92-fold and the LD 50 value increased by 100.22 to 3787.08 ng fly -1 after selection experiments. Moreover, the increase in LD 50 value remained stable when the selected strain (Perm-SEL) was cultured in the absence of permethrin for the next five generations (G11-G15). The results revealed the possibility of minor cross-resistance in the Perm-SEL strain with β-cyfluthrin and deltamethrin, and lack of cross-resistance with imidacloprid and spinosad. The realized heritability (h 2 ) value for the Perm-SEL strain was 0.23. Assuming a mean slope value of 1.27 and an h 2 value of 0.23, then ten to four generations would be estimated for a 10-fold increase in the LD 50 values at 50-90% selection intensity, respectively., Conclusion: The results of present study revealed high risk for permethrin resistance development under continuous selection pressure. Permethrin resistance developed very quickly under laboratory conditions, which might reflect the likelihood of resistance evolution in field conditions if permethrin selection pressure holds for a long time. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Background: Increasing resistance of head lice against neurotoxic agents and safety concerns have led to the search for treatment alternatives. Dimeticones with a physical mode of action are safe, and bear a reduced risk for the development of resistance., Methods: We performed in vitro bioassays to assess pediculicidal and ovicidal activities of a new dimeticone-based product, and a randomized controlled clinical trial to assess efficacy, following 10 min application. Of 153 individuals screened, 100 participants with active head louse infestations were randomly assigned to treatment with either a dimeticone-based test product, or a 0.5% permethrin-based reference product (50 participants per group). Participants received two topical applications of either the test (10 min) or reference products (45 min) at days 0 and 7 or 8. Outcome measures included the efficacies of treatment and their safety, as well as global and local tolerability at baseline, and days 1, 7, and 10., Results: After 10 min exposure, all lice treated with the dimeticone test product were classified as non-viable in the in vitro assay. Ovicidal activity after treatment of eggs with the dimeticone test product was 96.8%. In the clinical trial, 96 patients completed all study visits. In the full analysis set (FAS) population, on day 1 after one application, 98% of patients were cured in the test group, as compared to 84% cured in the reference group. All participants in both groups were free of head lice on day 10, following two applications (100% cure rate). In total, 42 adverse events (AEs) in 23 patients of both treatment groups were recorded, with the majority of AEs classified as mild., Conclusions: We have shown a high level of pediculicidal and ovicidal activity, and clinical efficacy and safety, of a brief application of a new dimeticone-based product. The short application time and reduced risk for the development of resistance are key drivers for improved patients' compliance., Trial Registration: EU Clinical Trials Register EudraCT  2016-004635-20 . Registered 14 November 2016.

Background: The brown dog tick, Rhipicephalus sanguineus sensu lato (Latreille; Acari: Ixodidae), is a peridomestic ectoparasite of dogs and occasionally humans. In some populations, lack of integrated pest management practices and overuse of pesticides has resulted in high levels of resistance to multiple active ingredients. In this study, we established the etofenprox discriminating concentration (DC) and three additional screening concentrations to evaluate resistance status. Using mortality results, cross-resistance was investigated in brown dog tick populations from five geographically disparate regions, including Asia, Africa, Europe, North America, and the Caribbean, following exposure to the DC for both etofenprox and permethrin separately. Subsequently, using both larval packet tests and molecular methods, etofenprox resistance was investigated., Results: The etofenprox DC was set at 0.51%, which allows for the rapid screening of peridomestic brown dog tick populations for resistance to this active ingredient. Cross-tolerance to two sodium channel-inhibiting pesticides, permethrin and etofenprox, was observed in one population., Conclusion: Our study has provided a single etofenprox concentration that can be used in larval packet tests to determine resistance status in collected peridomestic brown dog ticks, which is particularly important when a single to a few engorged adult ticks are provided for evaluation. Although metabolic resistance is presumed to be the primary resistance mechanism, a sodium channel mutation also confers tolerance to etofenprox at the DC. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Background: The highly invasive mosquito species Aedes albopictus has become a major health concern in temperate areas due to its role as vector of exotic arboviruses. Pyrethroid insecticides represent the main tools for limiting the circulation of such mosquito-borne viruses. The present work aim to extend previous reports on phenotypic pyrethroid-resistance in European Ae. albopictus, to identify its genetic basis and to monitor the geographical distribution of resistant genotypes, with a particular focus on sites experiencing the 2017 chikungunya outbreak in Italy., Results: Bioassays, performed according to World Health Organization protocols, showed full susceptibility to deltamethrin (concentration = 0.05%) and varying levels of resistance to permethrin (0.75%) and/or α-cypermethrin (0.05%) across Italy, with highest levels in the core of the 2017 chikungunya outbreak. Partial genotyping of the VSSC gene revealed widespread distribution of V1016G mutation and confirmed its association with pyrethroid resistance., Conclusion: The results obtained show that the condition for the spread of pyrethroid resistance in Ae. albopictus in Europe exists under strong selective pressure due to intensive insecticide spraying to control exotic arbovirus outbreak or high levels of nuisance. The results draw attention to the need for an evidence-based implementation of mosquito nuisance control, taking insecticide resistance management into consideration. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

The use of insecticides such as deltamethrin, permethrin, and temephos has been a primary tool to manage Aedes aegypti (Linnaeus) in Punjab province, Pakistan; however, recent reports of control failure necessitate monitoring insecticide resistance. For this reason, we evaluated 12 field strains of Ae. aegypti from Punjab for resistance against deltamethrin, permethrin, and temephos along with underlying resistance mechanisms. For deltamethrin, high level of resistance was observed in Rawalpindi, Faisalabad, Sheikhupura, Lahore, Pattoki, and Kasur strains (RRLC50 > 10-fold); moderate level of resistance in Sargodha, Gujranwala, and Sialkot strains (RRLC50 = 5- to 10-fold), and low level of resistance in Okara, Multan, and Sahiwal strains (RRLC50 < 5-fold). In the case of permethrin, high level of resistance was found in all the field strains, except the Okara strain that exhibited moderate resistance. For temephos, five field strains, viz. Faisalabad, Rawalpindi, Kasur, Lahore, and Gujranwala, showed high level of resistance; five strains, viz. Sheikhupura, Sialkot, Pattoki, Sahiwal, and Okara, showed moderate resistance, and two strains from Multan and Sargodha showed low resistance to temephos. Synergism bioassays implementing piperonyl butoxide and S,S,S-tributylphosphorotrithioate exhibited a nonsignificant effect on synergizing toxicity of deltamethrin and permethrin in all field strains except the Lahore strain, suggesting the possible role of target-site resistance mechanism. However, both synergists had a significant effect on synergizing toxicity of temephos in all field strains, suggesting the possibility of metabolic-based mechanism of insecticide resistance. In conclusion, the study confirmed the presence of resistance to deltamethrin, permethrin, and temephos in the studied field strains of Ae. aegypti from Punjab, Pakistan., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The rising threat of vector-borne diseases and environmental pollution has instigated the investigation of nanotechnology-based applications. The current study deals with a nanotechnological application involving the usage of nanometric pesticides such as permethrin nanoemulsion. The mean droplet diameter and zeta potential of the prepared permethrin nanoemulsion were found to be 12.4 ± 1.13 nm and -20.4 ± 0.56 mV, respectively. The temporal stability of permethrin nanoemulsion was found to be 4 days when checked in the external environment. The permethrin nanoemulsion exhibited LC 50 values of 0.038 and 0.047 mgL-1 and 0.049 and 0.063 mgL -1 against larval and pupal stages of Culex tritaeniorhynchus and Aedes aegypti, respectively. The results obtained from the larvicidal and pupicidal assay were corroborated with the histopathological and biochemical profiles of hosts upon treatment with nanometric pesticide. Further, the biosafety studies of the nanopesticide were carried out against different non-target species like freshwater algae (Closterium), Cicer arietinum (Chickpea) and Danio rerio (Zebrafish), and the mosquitocidal concentration of nanopesticide was found to be non-toxic. The following study, therefore, describes the mosquitocidal efficacy of nanometric pesticide formulated in a greener approach, which can become a substitute for conventional pesticide application in an eco-benign manner., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Resistance to insecticides can hamper the control of mosquitoes such as Culex quinquefasciatus, known to vector arboviruses such as West Nile virus and others. The strong selective pressure exerted on a mosquito population by the use of insecticides can result in heritable genetic changes associated with resistance. We sought to characterize genetic differences between insecticide resistant and susceptible Culex quinquefasciatus mosquitoes using targeted DNA sequencing. To that end, we developed a panel of 122 genes known or hypothesized to be involved in insecticide resistance, and used an Ion Torrent PGM sequencer to sequence 125 unrelated individuals from seven populations in the southern U.S. whose resistance phenotypes to permethrin and malathion were known from previous CDC bottle bioassay testing. Data analysis consisted of discovering SNPs (Single Nucleotide Polymorphism) and genes with evidence of copy number variants (CNVs) statistically associated with resistance. Ten of the seventeen genes found to be present in higher copy numbers were experimentally validated with real-time PCR. Of those, six, including the gene with the knock-down resistance (kdr) mutation, showed evidence of a ≥ 1.5 fold increase compared to control DNA. The SNP analysis revealed 228 unique SNPs that had significant p-values for both a Fisher's Exact Test and the Cochran-Armitage Test for Trend. We calculated the population frequency for each of the 64 nonsynonymous SNPs in this group. Several genes not previously well characterized represent potential candidates for diagnostic assays when further validation is conducted., Competing Interests: The authors have declared that no competing interests exist.

Insecticide-treated nets (ITNs) remain major components for vector control despite the spread of resistance mechanisms among mosquito populations. Multiple exposures to pyrethroids may induce physiological and behavioral changes in mosquitoes, possibly reducing efficacy of control tools. Despite epidemiological relevance, the effects of multiple exposures to pyrethroids on their efficacy against pyrethroid-resistant mosquitoes has received little interest. In the present study, we assessed the effects of a blood-meal successfully obtained upon a permethrin-treated net on the success at taking a second blood-meal in presence of permethrin in Anopheles gambiae, carrying pyrethroid resistance alleles. We also measured the impact of exposure to permethrin on life-history traits to address the delayed efficacy of ITNs. Our results showed that females that successfully blood-fed upon a permethrin-treated net were no longer inhibited by permethrin at the following exposure. Blood-meal inhibition due to permethrin was not affected by female size nor by exposure of mothers when testing the offspring, allowing to discard the effect of genetic or physiological selection. Besides, in our assays, exposure to permethrin did not affect mosquito fecundity, fertility nor survival. These results give insights to understand the long-term efficacy of ITNs, and allow to reevaluate the criteria used when choosing compounds for fighting malaria mosquitoes.

Human infestation with head lice, Pediculus humanus capitis De Geer, is the most prevalent ectoparasitic condition in the modern world. The purpose of this study was to test human head lice from Madagascar for infection with 2 louse-borne bacteria, Bartonella quintana and Acinetobacter spp. including Acinetobacter baumannii , to assess the potential risk of exposure to these pathogens in rural populations experiencing head-louse pediculosis. A second aim was to determine the occurrence of a biomarker for permethrin resistance in head lice from 6 isolated human communities in Madagascar. Deoxyribonucleic acid (DNA) of B. quintana was detected using species-specific Fab3 gene TaqMan in 12.6% of lice from 4 villages. DNA of Acinetobacter spp. was detected using rpo B TaqMan in 42.1% of lice collected from all locations; 58.3% of rpo B-positive lice had the bla OXA51 -like enzyme gene specific for A. baumannii . The kdr -resistant allele was detected in 70% of lice tested and was found in lice from each location. These results provide the first information regarding these combined characteristics of head-louse infestations in Madagascar. This approach can be applied to larger and broader surveys of lice from pediculosis capitis occurring in other geographic locations.

Background: Insecticide-treated bed nets (ITNs) help to control malaria by mechanically impeding the biting of mosquitoes, by repelling and irritating them and by killing them. In contrast to spatial repellency, irritancy implies that mosquitoes contact the ITN and are exposed to at least a sub-lethal dose of insecticide, which impedes their further blood-seeking. This would weaken the transmission of malaria, if mosquitoes are infectious., Methods: It was therefore tested whether sub-lethal exposure to permethrin impedes blood-feeding differently in uninfected mosquitoes and in mosquitoes carrying the non-transmissible stage (oocysts) or the infectious stage (sporozoites) of the malaria parasite Plasmodium berghei. In addition, as the degree of irritancy determines the dose of insecticide the mosquitoes may receive, the irritancy to permethrin of infected and uninfected mosquitoes was compared., Results: In this laboratory setting, sub-lethal exposure to permethrin inhibited the blood-seeking behaviour of Anopheles gambiae mosquitoes for almost 48 h. Although infection by malaria did not affect the irritancy of the mosquitoes to permethrin at either the developmental stage or the infectious stage, both stages of infection shortened the duration of inhibition of blood-seeking., Conclusions: The results suggest that the impact of ITNs may be weaker for malaria-infected than for uninfected mosquitoes. This will help to understand the global impact of ITNs on the transmission of malaria and gives a more complete picture of the effectiveness of that vector control measure.

A 2-year longitudinal study of enzyme-linked immunosorbent assay (ELISA) antibodies against Phlebotomus perniciosus and Phlebotomus papatasi (Diptera: Psychodidae) sandfly saliva was performed in 32 Beagle dogs treated preventively with an imidacloprid-permethrin topical insecticide in an endemic area in Spain. Dogs were grouped into three sandfly exposure groups according to the time of inclusion in the study. Assays analysed immunoglobulin G (IgG) against salivary gland homogenates (SGH) of both species and recombinant P. papatasi rSP32 and P. perniciosus rSP03B proteins in serum. The dogs were participating in a Leishmania infantum (Kinetoplastida: Trypanosomatidae) vaccine trial and were experimentally infected with the parasite in the second year. No dog acquired natural L. infantum infections during the first year, but most developed anti-saliva antibodies, and median log-transformed optical densities (LODs) were seasonal, mimicking those of local sandflies. This indicates that the repellent efficacy of the insecticide used is below 100%. Multi-level modelling of LODs revealed variability among dogs, autocorrelation and differences according to the salivary antigen and the dog's age. However, dog seroprevalence, estimated using pre-exposure LODs as cut-offs, was relatively low. This, and the fact that dogs did not become naturally infected with L. infantum, would support the efficacy and usefulness of this imidacloprid-permethrin topical insecticide in canine leishmaniasis control., (© 2018 The Royal Entomological Society.)

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 diabetes, which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid metabolism. The effects of 12 insecticides on triglycerides (TG) accumulation were tested using palmitic acid (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with palmitic acid to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty acid synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates fatty acid oxidation. Results in this study suggested that hepatic lipid metabolism may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD., (Copyright © 2019 Elsevier Ltd. All rights reserved.)