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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.

Background Leishmaniosis caused by Leishmania infantum is one of the most important vector-borne diseases affecting animals and humans worldwide. Dogs are considered main reservoirs of the zoonotic forms, though in the last years the role of cats as reservoirs has been increasingly investigated. Feline leishmaniosis (FeL) occurs in endemic areas and no specific preventive measures have been investigated so far. In this study the efficacy of a 10% imidacloprid/4.5% flumethrin polymer matrix collar, licensed for tick and flea prevention, has been assessed against FeL in a longitudinal study on 204 privately owned cats from the Aeolian islands (Sicily), an area highly endemic for the disease. From March to May 2015 [Study Day 0 (SD 0)], cats negative for FeL were collared (G1, n = 104) or left untreated (G2, n = 100). Diagnosis consisted of serology and qPCR on blood and conjunctival swabs, which were collected at baseline (SD 0) and at the end of the study (SD 360). Interim clinical examinations were performed on SD 210 (when collars were replaced in G1) and SD 270. Results Of the 159 cats which completed the study, 5 in G1 and 20 in G2 were positive for L. infantum infection, in at least one of the diagnostic tests leading to a yearly crude incidence of 6.3% and 25.0% in G1 and G2, respectively (P = 0.0026). This translates into an efficacy of the collar of 75.0% in preventing feline Leishmania infection. The collar was generally well tolerated with no systemic adverse reactions and few local skin reactions were observed in the application area in four out of 104 treated cats (3.8%). Conclusions The 10% imidacloprid/4.5% flumethrin collar significantly reduced the risk of L. infantum infection in cats. To our knowledge, this is the first study in which a preventative strategy against feline Leishmania infection is assessed under natural conditions. These findings close a gap in veterinary medicine, in that they confirm this collar as a tool in reducing the risk of Leishmania infection in cats. Such a preventative tool could contribute to the reduction of the risk of the disease in animal and in human populations when included in integrated leishmaniosis control programmes.

Background There is a paucity of data about the susceptibility status of malaria vectors to Public Health insecticides along the Thailand-Myanmar border. This lack of data is a limitation to guide malaria vector-control in this region. The aim of this study was to assess the susceptibility status of malaria vectors to deltamethrin, permethrin and DDT and to validate a simple molecular assay for the detection of knock-down resistance (kdr) mutations in the study area. Methods Anopheles mosquitoes were collected in four sentinel villages during August and November 2014 and July 2015 using human landing catch and cow bait collection methods. WHO susceptibility tests were carried out to measure the mortality and knock-down rates of female mosquitoes to deltamethrin (0.05%), permethrin (0.75%) and DDT (4%). DNA sequencing of a fragment of the voltage-gated sodium channel gene was carried out to identify knock-down resistance (kdr) mutations at position 1014 in mosquitoes surviving exposure to insecticides. Results A total of 6295 Anopheles belonging to ten different species were bioassayed. Resistance or suspected resistance to pyrethroids was detected in An. barbirostris (s.l.) (72 and 84% mortality to deltamethrin (n = 504) and permethrin (n = 493) respectively), An. hyrcanus (s.l.) (33 and 48% mortality to deltamethrin (n = 172) and permethrin (n = 154), respectively), An. jamesii (87% mortality to deltamethrin, n = 111), An. maculatus (s.l.) (85 and 97% mortality to deltamethrin (n = 280) and permethrin (n = 264), respectively), An. minimus (s.l.) (92% mortality, n = 370) and An. vagus (75 and 95% mortality to deltamethrin (n =148) and permethrin (n = 178), respectively). Resistance or suspected resistance to DDT was detected in An. barbirostris (s.l.) (74% mortality, n = 435), An. hyrcanus (s.l.) (57% mortality, n = 91) and An. vagus (97% mortality, n = 133). The L1014S kdr mutation at both heterozygous and homozygous state was detected only in An. peditaeniatus (Hyrcanus Group). Conclusion Resistance to pyrethroids is present along the Thailand-Myanmar border, and it represents a threat for malaria vector control. Further investigations are needed to better understand the molecular basis of insecticide resistance in malaria vectors in this area.

Human arboviral diseases transmitted by Aedes aegypti such as dengue, Zika, chikungunya and yellow fever remain global public health threats to date. Of these diseases, dengue fever is particularly prevalent in Southeast Asia. Relentless vector control efforts are performed to curtail disease transmissions through which pyrethroid insecticides are broadly used as the first line of defense to control Ae. aegypti, especially in the course of disease outbreaks. Here, we compile the largest contemporary database for susceptibility profiles and underlying mechanisms involved in Ae. aegypti resistant to pyrethroids in Southeast Asia. The extensive use of pyrethroids inevitably elicit different levels of resistance to numerous populations despite the presence of geographical isolation. The most common mechanisms of pyrethroid resistance that have been identified in Ae. aegypti includes mutations in the voltage sensitive sodium channel gene (Vssc gene) and metabolic-mediated insecticide resistance. Aedes aegypti develops resistance to pyrethroids by acquisition of one or several amino acid substitution(s) in this Vssc gene. Enzymes involved in metabolic-mediated detoxification (i.e. monooxygenases, glutathione-S-transferases and esterases) have been reported to be related to pyrethroid resistance but many specific contributory enzymes are not completely studied. An inadequate amount of data from some countries indicates an urgent need for further study to fill the knowledge gaps. Perspectives and future research needs are also discussed.

International audience; Background: Mauritania is one of the African countries with ongoing malaria transmission where data on insecticide resistance of local malaria vectors are limited despite an increasing use of long-lasting insecticide-treated nets (LLINs) as the main intervention for vector control. This study presents an evaluation of the level of insecticide resistance of Anopheles arabiensis in Nouakchott. Methods: Anopheles gambiae (s.l.) larvae were collected in breeding sites during the rainy season (August-September) in 2015 and 2016 from two selected sites in Nouakchott and reared until emergence. Adult anopheline mosquitoes were tested against malathion (5%), bendiocarb (0.1%), permethrin (0.75%) and deltamethrin (0.05%) using standard World Health Organization (WHO) insecticide-impregnated papers. PCR assays were used for the identification of An. gambiae (s.l.) sibling species as well as knockdown resistance (kdr). Results: The mean knockdown times 50% (KDT50) and 95% (KDT95) were 66 +/- 17 and 244 +/- 13 min, respectively, for permethrin in 2015. The KDT50 and the KDT95 were 39 +/- 13 and 119 +/- 13 min, respectively, for deltamethrin. The KDT50 and the KDT95 doubled for both molecules in 2016. The mortality rates 24 h post-exposure revealed that An. arabiensis populations in Nouakchott were fully susceptible to bendiocarb and malathion in 2015 as well as in 2016, while they were resistant to permethrin (51.9% mortality in 2015 and 24.1% mortality in 2016) and to deltamethrin (83.7% mortality in 2015 and 39.1% mortality in 2016). The molecular identification showed that Anopheles arabiensis was the only malaria vector species collected in Nouakchott in 2015 and 2016. Both the West and East African kdr mutant alleles were found in An. arabiensis mosquitoes surviving exposure to pyrethroid insecticide, with a high rate of homozygous resistant genotypes (54.3% for the West African kdr mutation and 21.4% for the East African kdr mutation) and a significant departure from Hardy-Weinberg proportions (chi(2) 134, df = 3, P < 0.001). Conclusions: The study showed high levels of pyrethroid resistance in An. arabiensis populations in Nouakchott and presence of both West and East African kdr alleles in the resistant phenotype. These results highlight a need for routine monitoring of susceptibility of malaria vector populations to insecticides used in public health programs.

Background High coverage of long-lasting insecticidal nets (LLINs) is the cornerstone of the malaria control strategy of the national malaria control program (NMCP) in Cameroon, with a target of reducing malaria transmission to less than 10% by 2035. To this end, more than 20 million LLINs have been distributed to populations countrywide since 2011. The present study evaluated entomological indices and Anopheles susceptibility to pyrethroids in a rural forested area of south Cameroon with high coverage of LLINs. Methods The study was conducted between July 2014 and May 2016 in Obout, a village located in a rural forested area in south Cameroon. Resting mosquitoes were collected using electric aspirators and were identified to species using morphological criteria and PCR tools. Mosquito feeding preferences and infection status to Plasmodium falciparum were determined by ELISA and using TaqMan assays. The susceptibility of wild F1 adults to pyrethroids was monitored using WHO insecticide susceptibility bioassays. Results During the study period, 5,993 Anopheles mosquitoes were collected indoors both in rooms with and without nets. Two main vector species, namely An. funestus and An. gambiae, were identified in the locality, with An. funestus being by far the most abundant (89.68%). ELISA analysis revealed high percentage of blood meal taken exclusively on human (97.65–98.95%) supporting the high antropohilic behaviour of both species. Plasmodium falciparum infection rate detected by ELISA was high throughout the study period and varied between 3.28–14.04% (mean: 10.40%) in An. funestus, and between 5.55–22.22% (mean: 13.87%) in An. gambiae. This trend was confirmed by TaqMan assays, with P. falciparum infection prevalence of 23.33% in An. funestus. Significant decrease of mortality associated with high frequency of kdr mutation was observed in An. gambiae (deltamethrin: 36.6–56.45%; permethrin: 6–18.65%) indicating high level of resistance to pyrethroids. For An. funestus, resistance was marked for deltamethrin (mortality: 70.54–76.24%) than for permethrin (94.12–94.74%). Conclusions Our study showed that despite LLINs, the population of Obout remains exposed to bites of highly infected An. funestus and An. gambiae mosquitoes, highlighting the challenges to controlling malaria in forested areas, especially in the presence of insecticide resistance.

Background Metofluthrin reduces biting activity in Aedes aegypti through the confusion, knockdown, and subsequent kill of a mosquito. A geographical spread in dengue, chikungunya, and Zika viruses, increases intervention demands. Response to a Zika outbreak may require a different strategy than dengue, as high-risk individuals, specifically pregnant women, need to be targeted. Methods In semi-field conditions within a residential property in Cairns, Queensland, the impacts of metofluthrin on biting behaviour of free-flying Wolbachia-infected Ae. aegypti were evaluated. Results Mortality in Ae. aegypti exposed to metofluthrin over a 22 h period was 100% compared to 2.7% in an untreated room. No biting activity was observed in mosquitoes up to 5 m from the emanator after 10 min of metofluthrin exposure. Use of metofluthrin reduced biting activity up to 8 m, regardless of the host’s proximity (near or far) to a dark harbourage area (HA) (P

Background Pyrethroid resistance in African vector mosquitoes is a threat to malaria control. Resistant mosquitoes can survive insecticide doses that would normally be lethal. We studied effects of such doses on Plasmodium falciparum development inside kdr-resistant Anopheles gambiae s.s. in Uganda. Methods We collected An. gambiae s.s. homozygous for kdr-L1014S mutation, fed them on blood samples from 42 P. falciparum-infected local patients, then exposed them either to nets treated with sub-lethal doses of deltamethrin or to untreated nets. After seven days, we dissected 692 mosquitoes and examined their midguts for oocysts. Prevalence (proportion infected) and intensity of infection (number of oocysts per infected mosquito) were recorded for each group. Results Both prevalence and intensity of infection were significantly reduced in deltamethrin-exposed mosquitoes, compared to those exposed to untreated nets. With low doses (2.5–5.0 mg/m2), prevalence was reduced by 59 % (95 % CI = 22 %-78 %) and intensity by 41 % (95 % CI = 25 %-54 %). With high doses (10–16.7 mg/m2), prevalence was reduced by 80 % (95 % CI = 67 %-88 %) and intensity by 34 % (95 % CI = 20 %-46 %). Conclusions We showed that, with locally-sampled parasites and mosquitoes, doses of pyrethroids that are sub-lethal for resistant mosquitoes can interfere with parasite development inside mosquitoes. This mechanism could enable pyrethroid-treated nets to prevent malaria transmission despite increasing vector resistance.

Background Sarcoptes scabiei infection is a contagious disease affecting both humans and animals. The transmission occurs either by direct contact or from the environment where mites could survive several days remaining infective. The number of products available for environmental control of S. scabiei is very limited. The objective of the present study was to assess the efficacy of biocides or repellents against S. scabiei var suis. Methods Tested products included pyrethroids: permethrin, esdepallethrin and bioresmethrin, bifenthrin, cypermethrin and imiprothrin, cyfluthrin, tetramethrin and sumithrin. We also tested repellents: DEET, icaridin and IR3535. Sarcoptes scabiei var suis mites were collected from experimentally-infected pigs. For each test, 20 live mites of all motile stages were placed in a plastic Petri dish and sprayed uniformly by each product. Control mites were sprayed by distilled water. The study was performed in triplicate under room conditions and the mites were inspected under a stereomicroscope at intervals (5, 10, 15, 20, 25, 30, 40, 50, 60 min, 2, 3, 4, 5 and 24 h) after exposure to the products. Results All the products, except the combination of tetramethrin and sumithrin (A-PAR®), were able to kill all mites within 24 h. The median survival time was 50 ± 30.4 min, 120 ± 309 min, 10 ± 5.9 min, 40 ± 36.8 min, 15 ± 7.3 min, 180 ± 417 min and 1440 ± 600 min when mites were exposed to permethrin 4 %, permethrin 0.6 %, esdepallethrin and bioresmethrin, bifenthrin, cypermethrin and imiprothrin, cyfluthrin, tetramethrin and sumithrin, respectively. The median survival time was 20 ± 6.5 min, 15 ± 4.3 min, 30 ± 42.1 min and 15 ± 4.9 min for DEET 25, DEET 50, icaridin 20 and IR3535 20 %, respectively. Conclusions The results of the present study could support evidence-based use of biocides and repellents in households, hospitals and farms.

Background Mosquito control based on chemical insecticides is considered as an important element in the current global strategies for the control of mosquito-borne diseases. Unfortunately, the development of pyrethroid resistance in important vector mosquito species jeopardizes the effectiveness of insecticide-based mosquito control. To date, the mechanisms of pyrethroid resistance are still unclear. Recent advances in proteomic techniques can facilitate to identify pyrethroid resistance-associated proteins at a large-scale for improving our understanding of resistance mechanisms, and more importantly, for seeking some genetic markers used for monitoring and predicting the development of resistance. Methods We performed a quantitative proteomic analysis between a deltamethrin-susceptible strain and a deltamethrin-resistant strain of laboratory population of Culex pipiens pallens using isobaric tags for relative and absolute quantitation (iTRAQ) analysis. Gene Ontology (GO) analysis was used to find the relative processes that these differentially expressed proteins were involved in. One differentially expressed protein was chosen to confirm by Western blot in the laboratory and field populations of Cx. pipiens pallens. Results We identified 30 differentially expressed proteins assigned into 10 different categories, including oxidoreductase activity, transporter activity, catalytic activity, structural constituent of cuticle and hypothetical proteins. GO analysis revealed that 25 proteins were sub-categorized into 35 hierarchically-structured GO classifications. Western blot results showed that CYP6AA9 as one of the up-regulated proteins was confirmed to be overexpressed in the deltamethrin-resistant strains compared with the deltamethrin-susceptible strains both in the laboratory and field populations. Conclusions This is the first study to use modern proteomic tools for identifying pyrethroid resistance-related proteins in Cx. pipiens. The present study brought to light many proteins that were not previously thought to be associated with pyrethroid resistance, which further expands our understanding of pyrethroid resistance mechanisms. CYP6AA9 was overexpressed in the deltamethrin-resistant strains, indicating that CYP6AA9 may be involved in pyrethroid resistance and may be used as a potential genetic marker to monitor and predict the pyrethroid resistance level of field populations. Electronic supplementary material The online version of this article (doi:10.1186/s13071-015-0709-5) contains supplementary material, which is available to authorized users.

Constant and extensive use of chemical insecticides has created a selection pressure and favored resistance development in many insect species worldwide. One of the most important pyrethroid resistance mechanisms is classified as target site insensitivity, due to conformational changes in the target site that impair a proper binding of the insecticide molecule. The voltage-gated sodium channel (NaV) is the target of pyrethroids and DDT insecticides, used to control insects of medical, agricultural and veterinary importance, such as anophelines. It has been reported that the presence of a few non-silent point mutations in the NaV gene are associated with pyrethroid resistance, termed as ‘kdr’ (knockdown resistance) for preventing the knockdown effect of these insecticides. The presence of these mutations, as well as their effects, has been thoroughly studied in Anopheles mosquitoes. So far, kdr mutations have already been detected in at least 13 species (Anopheles gambiae, Anopheles arabiensis, Anopheles sinensis, Anopheles stephensi, Anopheles subpictus, Anopheles sacharovi, Anopheles culicifacies, Anopheles sundaicus, Anopheles aconitus, Anopheles vagus, Anopheles paraliae, Anopheles peditaeniatus and Anopheles albimanus) from populations of African, Asian and, more recently, American continents. Seven mutational variants (L1014F, L1014S, L1014C, L1014W, N1013S, N1575Y and V1010L) were described, with the highest prevalence of L1014F, which occurs at the 1014 site in NaV IIS6 domain. The increase of frequency and distribution of kdr mutations clearly shows the importance of this mechanism in the process of pyrethroid resistance. In this sense, several species-specific and highly sensitive methods have been designed in order to genotype individual mosquitoes for kdr in large scale, which may serve as important tolls for monitoring the dynamics of pyrethroid resistance in natural populations. We also briefly discuss investigations concerning the course of Plasmodium infection in kdr individuals. Considering the limitation of insecticides available for employment in public health campaigns and the absence of a vaccine able to brake the life cycle of the malaria parasites, the use of pyrethroids is likely to remain as the main strategy against mosquitoes by either indoor residual spraying (IR) and insecticide treated nets (ITN). Therefore, monitoring insecticide resistance programs is a crucial need in malaria endemic countries. Electronic supplementary material The online version of this article (doi:10.1186/1756-3305-7-450) contains supplementary material, which is available to authorized users.

BACKGROUND: Increasing pyrethroid resistance in malaria vectors has been reported in western Kenya where long lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) are the mainstays of vector control. To ensure the sustainability of insecticide-based malaria vector control, monitoring programs need to be implemented. This study was designed to investigate the extent and distribution of pyrethroid resistance in 4 Districts of western Kenya (Nyando, Rachuonyo, Bondo and Teso). All four Districts have received LLINs while Nyando and Rachuonyo Districts have had IRS campaigns for 3-5 years using pyrethroids. This study is part of a programme aimed at determining the impact of insecticide resistance on malaria epidemiology. METHODS: Three day old adult mosquitoes from larval samples collected in the field, were used for bioassays using the WHO tube bioassay, and mortality recorded 24 hours post exposure. Resistance level was assigned based on the 2013 WHO guidelines where populations with

INTRODUCTION:\ud The voltage gated sodium channel mutation Vgsc-1014S (kdr-east) was first reported in Kenya in 2000 and has since been observed to occur at high frequencies in the local Anopheles gambiae s.s.\ud POPULATION:\ud The mutation Vgsc-1014F has never been reported from An. gambiae Complex complex mosquitoes in Kenya.\ud FINDINGS:\ud Molecularly confirmed An. gambiae s.s. (hereafter An. gambiae) and An. arabiensis collected from 4 different parts of western Kenya were genotyped for kdr from 2011 to 2013. Vgsc-1014F was observed to have emerged, apparently, simultaneously in both An. gambiae and An. arabiensis in 2012. A portion of the samples were submitted for sequencing in order to confirm the Vgsc-1014F genotyping results. The resulting sequence data were deposited in GenBank (Accession numbers: KR867642-KR867651, KT758295-KT758303). A single Vgsc-1014F haplotype was observed suggesting, a common origin in both species.\ud CONCLUSION:\ud This is the first report of Vgsc-1014F in Kenya. Based on our samples, the mutation is present in low frequencies in both An. gambiae and An. arabiensis. It is important that we start monitoring relative frequencies of the two kdr genes so that we can determine their relative importance in an area of high insecticide treated net ownership.

Background Mosquitoes host and pass on to humans a variety of disease-causing pathogens such as infectious viruses and other parasitic microorganisms. The emergence and spread of insecticide resistance is threatening the effectiveness of current control measures for common mosquito vector borne diseases, such as malaria, dengue and Zika. Therefore, the emerging resistance to the widely used pyrethroid insecticides is an alarming problem for public health. Herein we demonstrated the use of RNA interference (RNAi) to increase susceptibility of adult mosquitoes to a widely used pyrethroid insecticide. Methods Experiments were performed on a field-collected pyrethroid resistant strain of Ae. aegypti (Rio de Janeiro; RJ). Larvae from the resistant Ae. aegypti population were soaked with double-stranded RNAs (dsRNAs) that correspond either to voltage-gate sodium channel (VGSC), P-glycoprotein, or P450 detoxification genes and reared to adulthood. Adult mortality rates in the presence of various Deltamethrin pyrethroid concentrations were used to assess mosquito insecticide susceptibility. Results We characterized the RJ Ae. aegypti strain with regard to its level of resistance to a pyrethroid insecticide and found that it was approximately 6 times more resistant to Deltamethrin compared to the laboratory Rockefeller strain. The RJ strain displayed a higher frequency of Val1016Ile and Phe1534Cys substitutions of the VGSC gene. The resistant strain also displayed a higher basal expression level of VGSC compared to the Rockefeller strain. When dsRNA-treated mosquitoes were subjected to a standard pyrethroid contact bioassay, only dsRNA targeting VGSC increased the adult mortality of the pyrethroid resistant strain. The dsRNA treatment proved effective in increasing adult mosquito susceptibility over a range of pyrethroid concentrations and these results were associated with dsRNA-specific small interfering RNAs in treated adults, and the corresponding specific down regulation of VGSC gene expression level. Finally, we demonstrated that the efficiency of our approach was further improved by ‘tiling’ along the VGSC gene in order to identify the most potent dsRNA sequences. Conclusions These results demonstrate that dsRNA applied to mosquito larvae retains its biological activity into adulthood. Thus, the RNAi system reported here could be a useful approach to control the widespread insecticide resistance in mosquitoes and other insect vectors of human diseases. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1634-y) contains supplementary material, which is available to authorized users.