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The landscape of mosquito-borne disease risk has changed dramatically in recent decades, due to the emergence and reemergence of urban transmission cycles driven by invasive Aedes aegypti and Ae. albopictus. Insecticide resistance is already widespread in the yellow fever mosquito, Ae. Aegypti; is emerging in the Asian tiger mosquito Ae. Albopictus; and is now threatening the global fight against human arboviral diseases such as dengue, yellow fever, chikungunya, and Zika. Because the panel of insecticides available for public health is limited, it is of primary importance to preserve the efficacy of existing and upcoming active ingredients. Timely implementation of insecticide resistance management (IRM) is crucial to maintain the arsenal of effective public health insecticides and sustain arbovirus vector control. This Review is one of a series being generated by the Worldwide Insecticide resistance Network (WIN) and aims at defining the principles and concepts underlying IRM, identifying the main factors affecting the evolution of resistance, and evaluating the value of existing tools for resistance monitoring. Based on the lessons taken from resistance strategies used for other vector species and agricultural pests, we propose a framework for the implementation of IRM strategies for Aedes mosquito vectors. Although IRM should be a fixture of all vector control programs, it is currently often absent from the strategic plans to control mosquito-borne diseases, especially arboviruses. Experiences from other public health disease vectors and agricultural pests underscore the need for urgent action in implementing IRM for invasive Aedes mosquitoes. Based on a plan developed for malaria vectors, here we propose some key activities to establish a global plan for IRM in Aedes spp.

Background Pyrethroid and organophosphate resistance in the malaria vector Anopheles gambiae has led to the search for not only alternative insecticides, but also repellent chemical compounds. However, little is known about the potential actions of repellents and the cross-resistance risk between insecticide and repellent compounds. Methods Here we show the action of permethrin, DEET, geraniol, carvacrol, culminaldehyde and cinnamaldehyde against three A. gambiae strains: ‘Kis’ (Kisumu susceptible strain), ‘KdrKis’ (pyrethroid resistant strain) and ‘AcerKis’ (organophosphate resistant strain), the last two differing from the first by a mutation on the kdr and ace1 genes, respectively. Conclusions Results from the DEET assays show it induced repellency for the resistant KdrKis and AcerKis strains but maintained irritancy for the susceptible strain. More generally, we show resistance genes modify the behavior of An. gambiae , increasing or decreasing the effectiveness of DEET and natural compounds, depending on the mutation. These findings offer a new avenue for research on the target and mechanism of repellent compounds. We discuss these findings in the context of vector control strategies.

To tackle the problem of insecticide resistance, all resistance mechanisms need to be studied. This study investigated the involvement of the cuticle in pyrethroid resistance in a strain of Anopheles gambiae , MRS, free of kdr mutations. Bioassays revealed MRS to be resistant to pyrethroids and DDT, indicated by increasing knockdown times and resistance ratios. Moreover, biochemical analysis indicated that metabolic resistance based on enhanced CYP450 activity may also play a role. Insecticide penetration assays showed that there were significantly lower amounts of insecticide in the MRS strain than in the susceptible control. Analysis of the levels of the selected transcripts by qPCR showed that CYP6M2, a major pyrethroid metaboliser, CYP4G16, a gene implicated in resistance via its contribution to the biosynthesis of elevated epicuticular hydrocarbons that delay insecticide uptake, and the cuticle genes CPAP3-E and CPLCX1 were upregulated after insecticide exposure. Other metabolic (CYP6P3, GSTe2) and cuticle (CPLCG3, CPRs) genes were also constitutively upregulated. Microscopic analysis showed that the cuticle layers of the MRS strain were significantly thicker than those of the susceptible strain. This study allowed us to assess the contribution made by the cuticle and metabolic mechanisms to pyrethroid resistance in Anopheles gambiae without target-site mutations.

Urban Aedes mosquitoes are vectors of many viruses affecting human health such as dengue, chikungunya and Zika viruses. Insecticide resistance and environmental toxicity risks hamper the effectiveness of chemical control against these mosquito vectors. Alternative control methods, such as the use of mosquito-specific entomopathogenic viruses should be explored. Numerous studies have focused on evaluating the potential of different densoviruses species as biological control agents. However, knowledge on the extent of inter- and intra-specific variations in the susceptibility of Aedes mosquitoes to infection by different densoviruses remains insufficient. In this study, we compared infection and mortality rates induced by the Aedes albopictus densovirus 2 in different strains of Aedes albopictus and Aedes aegypti mosquitoes. The two Aedes species were different in terms of susceptibility to viral infection. Under laboratory conditions, Aedes albopictus densovirus 2 appeared more virulent for the different strains of Aedes aegypti tested than for those of Aedes albopictus . In addition, we also found significant intra-specific variation in infection and mortality rates. Thus, although even if Aedes albopictus densoviruses could be powerful biocontrol agents used in the management of urban Aedes populations, our results also call into question the use of single viral isolate as biocontrol agents.

Background Carapa procera products, particularly the vegetable oil, are prized for their cosmetic, medicinal and especially insect repellent/insecticide properties. The latter property raises questions about the use of this plant against mosquitoes that transmit pathogens within Ivorian communities, given that Côte d’Ivoire is endemic for malaria. There is no scientific report on the use of C. procera and the plants traditionally used to prevent mosquito bites in Côte d’Ivoire. The aim of this study is therefore to determine the virtues of C. procera known to the people of Côte d’Ivoire and to identify the plants they use to protect themselves from mosquito bites. Methods A descriptive qualitative study was conducted in 16 localities in 6 regions of Côte d’Ivoire. The surveys were carried out using non-probability sampling and semi-structured interviews. The sample for each locality was selected on the basis of response saturation. Samples of available or accessible plants cited by local people were collected for identification in the herbarium. Results A total of 121 people were interviewed. The study showed that 68.6% of respondents were familiar with C. procera . Carapa procera vegetable oil has been designated as an insect repellent, particularly for flies, mosquitoes and ants. The cosmetic, antimalarial, anti-fatigue and mystical uses of the plant have been listed. The study showed that 47.1% of respondents used plants to protect themselves against mosquitoes. Twelve plant species traditionally used against mosquitoes were identified: Nicotiana tabacum, Hyptis suaveolens , Ocimum basilicum , Citrus sinensis, Azadirachta indica, Elaeis guineensis , Chromolaena odorata, Hoslundia opposita , Ocimum gratissimum , Piliostigma thonningii , Alstonia boone i and Phyllanthus amarus. The most common method of use was to fumigate fresh or dried plants inside homes. Conclusion The study presented the reasons for the use of C. procera and other plants used against mosquitoes in Côte d’Ivoire. It shows the importance of plant substances in protecting communities against mosquitoes, despite the deployment of long-lasting insecticidal bed nets (LLINs) and the spraying of insecticides by health programmes.

Dengue is an important mosquito borne viral disease in Martinique Island (French West Indies). The viruses responsible for dengue are transmitted by Aedes aegypti, an indoor day-biting mosquito. The most effective proven method for disease prevention has been by vector control by various chemical or biological means. Unfortunately insecticide resistance has already been observed on the Island and recently showed to significantly reduce the efficacy of vector control interventions. In this study, we investigated the distribution of resistance and the underlying mechanisms in nine Ae. aegypti populations. Statistical multifactorial approach was used to investigate the correlations between insecticide resistance levels, associated mechanisms and environmental factors characterizing the mosquito populations. Bioassays revealed high levels of resistance to temephos and deltamethrin and susceptibility to Bti in the 9 populations tested. Biochemical assays showed elevated detoxification enzyme activities of monooxygenases, carboxylesterases and glutathione S-tranferases in most of the populations. Molecular screening for common insecticide target-site mutations, revealed the presence of the \"knock-down resistance\" V1016I Kdr mutation at high frequency (>87%). Real time quantitative RT-PCR showed the potential involvement of several candidate detoxification genes in insecticide resistance. Principal Component Analysis (PCA) performed with variables characterizing Ae. aegypti from Martinique permitted to underline potential links existing between resistance distribution and other variables such as agriculture practices, vector control interventions and urbanization. Insecticide resistance is widespread but not homogeneously distributed across Martinique. The influence of environmental and operational factors on the evolution of the resistance and mechanisms are discussed.

Background Insecticide resistance monitoring is essential to help national programmers to implement more effective and sustainable malaria control strategies in endemic countries. This study reported the spatial and seasonal variations of insecticide resistance in malaria vectors in Benin, West Africa. Methods Anopheles gambiae s.l populations were collected from October 2008 to June 2010 in four sites selected on the basis of different use of insecticides and environment. WHO susceptibility tests were carried out to detect resistance to DDT, fenitrothion, bendiocarb, permethrin and deltamethrin. The synergist piperonyl butoxide was used to assess the role of non-target site mechanisms in pyrethroid resistance. Anopheles gambiae mosquitoes were identified to species and to molecular M and S forms using PCR techniques. Molecular and biochemical assays were carried out to determine kdr and Ace.1 R allelic frequencies and activity of the detoxification enzymes. Results Throughout the surveys very high levels of mortality to bendiocarb and fenitrothion were observed in An. gambiae s.l . populations. However, high frequencies of resistance to DDT and pyrethroids were seen in both M and S form of An. gambiae s.s . and Anopheles arabiensis . PBO increased the toxicity of permethrin and restored almost full susceptibility to deltamethrin. Anopheles gambiae s.l . mosquitoes from Cotonou and Malanville showed higher oxidase activity compared to the Kisumu susceptible strain in 2009, whereas the esterase activity was higher in the mosquitoes from Bohicon in both 2008 and 2009. A high frequency of 1014F kdr allele was initially showed in An. gambiae from Cotonou and Tori-Bossito whereas it increased in mosquitoes from Bohicon and Malanville during the second year. For the first time the L1014S kdr mutation was found in An. arabiensis in Benin. The ace.1 R mutation was almost absent in An. gambiae s.l . Conclusion Pyrethroid and DDT resistance is widespread in malaria vector in Benin and both metabolic and target site resistance are implicated. Resistance was not correlated with a change of malaria species and/or molecular forms. The 1014S kdr allele was first identified in wild population of An. arabiensis hence confirming the expansion of pyrethroid resistance alleles in Africa.

Aedes mosquitoes are the vectors of dengue viruses and other arboviruses, which threaten billions of people all over the world. The boosted sterile insect technique (boosted SIT) is a version of SIT in which irradiated sterile males also transmit a biocide to immature stages. We describe three field trials that were run in 2021: one against Aedes aegypti in La Reunion and two against Aedes albopictus in Spain, each using pyriproxyfen as a biocide. The relative density of adults (compared to their density in control sites: without sterile male release) decreased from 1.00 to 0.09, 95% credible interval [0.06, 0.15] (La Reunion, July) and to 0.02 [0.01, 0.03] and 0.11 [0.08, 0.16] (Spain, July and October). The success rate, corresponding to the proportion of traps with suppression greater than 80%, ranged from 0.43 to 0.71 in La Reunion, from 0.26 to 1.00, and from 0.50 to 0.70 in Spain. In Spain, suppression with boosted SIT was higher than with non-boosted SIT, in 2020 and 2022. This work is in line with the predictions of the model of a better efficacy of boosted SIT compared to SIT, together with partial protection from invasion of treated areas by fertile females, paving the way for larger-scale field trials.

Vector control is the mainstay of malaria control programmes. Successful vector control profoundly relies on accurate information on the target mosquito populations in order to choose the most appropriate intervention for a given mosquito species and to monitor its impact. An impediment to identify mosquito species is the existence of morphologically identical sibling species that play different roles in the transmission of pathogens and parasites. Currently PCR diagnostics are used to distinguish between sibling species. PCR based methods are, however, expensive, time-consuming and their development requires a priori DNA sequence information. Here, we evaluated an inexpensive molecular proteomics approach for Anopheles species: matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). MALDI-TOF MS is a well developed protein profiling tool for the identification of microorganisms but so far has received little attention as a diagnostic tool in entomology. We measured MS spectra from specimens of 32 laboratory colonies and 2 field populations representing 12 Anopheles species including the A. gambiae species complex. An important step in the study was the advancement and implementation of a bioinformatics approach improving the resolution over previously applied cluster analysis. Borrowing tools for linear discriminant analysis from genomics, MALDI-TOF MS accurately identified taxonomically closely related mosquito species, including the separation between the M and S molecular forms of A. gambiae sensu stricto. The approach also classifies specimens from different laboratory colonies; hence proving also very promising for its use in colony authentication as part of quality assurance in laboratory studies. While being exceptionally accurate and robust, MALDI-TOF MS has several advantages over other typing methods, including simple sample preparation and short processing time. As the method does not require DNA sequence information, data can also be reviewed at any later stage for diagnostic or functional patterns without the need for re-designing and re-processing biological material.

A shift towards early morning biting behavior of the major malaria vector Anopheles funestus have been observed in two villages in south Benin following distribution of long-lasting insecticidal nets (LLINs), but the impact of these changes on the personal protection efficacy of LLINs was not evaluated. Data from human and An. funestus behavioral surveys were used to measure the human exposure to An. funestus bites through previously described mathematical models. We estimated the personal protection efficacy provided by LLINs and the proportions of exposure to bite occurring indoors and/or in the early morning. Average personal protection provided by using of LLIN was high (≥80% of the total exposure to bite), but for LLIN users, a large part of remaining exposure occurred outdoors (45.1% in Tokoli-V and 68.7% in Lokohoué) and/or in the early morning (38.5% in Tokoli-V and 69.4% in Lokohoué). This study highlights the crucial role of LLIN use and the possible need to develop new vector control strategies targeting malaria vectors with outdoor and early morning biting behavior. This multidisciplinary approach that supplements entomology with social science and mathematical modeling illustrates just how important it is to assess where and when humans are actually exposed to malaria vectors before vector control program managers, policy-makers and funders conclude what entomological observations imply.