Chikungunya Virus Infection and Gonotrophic Cycle Shape Aedes aegypti Oviposition Behavior and Preferences.

Targeting gravid females through chemical lures is a promising strategy in vector control; however, it requires the understanding of the factors susceptible to alter female oviposition behavior. Here, we evaluated the effect of infection with chikungunya virus (CHIKV) and the number of gonotrophic cycles (GCs) on oviposition activity in A. aegypti. Dual choice oviposition assays were performed, where dodecanoic acid, pentadecanoic acid, n-heneicosane and a Sargasssum fluitans (Børgesen) Børgesen extract were tested in uninfected females and females infected with CHIKV, at the 1st and 2nd GC. Infected females displayed a lower percentage of oviposition and a higher number of eggs laid at the 1st GC. Then, the combined effects of GC and CHIKV were observed on oviposition preferences, with a chemical-dependent effect. For instance, the deterrent effect of n-heneicosane and pentadecanoic acid increased at the 2nd GC in infected females. These results allow for a deeper understanding of the mechanisms involved in oviposition site selection and highlight the need for taking into account physiological stage changes to increase the control programs' efficacy.

Semiochemical oviposition cues to control Aedes aegypti gravid females: state of the art and proposed framework for their validation.

In the fight against mosquito-borne diseases, odour-based lures targeting gravid females represent a promising alternative to conventional tools for both reducing mosquito populations and monitoring pathogen transmission. To be sustainable and effective, they are expected to use semiochemicals that act specifically against the targeted vector species. In control programmes directed against Aedes aegypti, several candidates of different origins (conspecifics, plants) have already been identified as potential oviposition attractants or repellents in laboratory experiments. However, few of these candidates have received validation in field experiments, studies depicting the active molecules and their mode of perception are still scarce, and there are several methodological challenges (i.e. lack of standardization, differences in oviposition index interpretation and use) that should be addressed to ensure a better reproducibility and accelerate the validation of candidates. In this review, we address the state of the art of the compounds identified as potential candidates for trap development against Ae. aegypti and their level of validation. We also offer a critical methodological analysis, highlight remaining gaps and research priorities, and propose a workflow to validate these candidates and to increase the panel of odours available to specifically trap Ae. aegypti.

Controlled human malaria infection with a clone of Plasmodium vivax with high-quality genome assembly.

Controlled human malaria infection (CHMI) provides a highly informative means to investigate host-pathogen interactions and enable in vivo proof-of-concept efficacy testing of new drugs and vaccines. However, unlike Plasmodium falciparum, well-characterized P. vivax parasites that are safe and suitable for use in modern CHMI models are limited. Here, 2 healthy malaria-naive United Kingdom adults with universal donor blood group were safely infected with a clone of P. vivax from Thailand by mosquito-bite CHMI. Parasitemia developed in both volunteers, and prior to treatment, each volunteer donated blood to produce a cryopreserved stabilate of infected RBCs. Following stringent safety screening, the parasite stabilate from one of these donors (PvW1) was thawed and used to inoculate 6 healthy malaria-naive United Kingdom adults by blood-stage CHMI, at 3 different dilutions. Parasitemia developed in all volunteers, who were then successfully drug treated. PvW1 parasite DNA was isolated and sequenced to produce a high-quality genome assembly by using a hybrid assembly method. We analyzed leading vaccine candidate antigens and multigene families, including the vivax interspersed repeat (VIR) genes, of which we identified 1145 in the PvW1 genome. Our genomic analysis will guide future assessment of candidate vaccines and drugs, as well as experimental medicine studies.

Mosquito Attractants.

Vector control and personal protection against anthropophilic mosquitoes mainly rely on the use of insecticides and repellents. The search for mosquito-attractive semiochemicals has been the subject of intense studies for decades, and new compounds or odor blends are regularly proposed as lures for odor-baited traps. We present a comprehensive and up-to-date review of all the studies that have evaluated the attractiveness of volatiles to mosquitoes, including individual chemical compounds, synthetic blends of compounds, or natural host or plant odors. A total of 388 studies were analysed, and our survey highlights the existence of 105 attractants (77 volatile compounds, 17 organism odors, and 11 synthetic blends) that have been proved effective in attracting one or several mosquito species. The exhaustive list of these attractants is presented in various tables, while the most common mosquito attractants - for which effective attractiveness has been demonstrated in numerous studies - are discussed throughout the text. The increasing knowledge on compounds attractive to mosquitoes may now serve as the basis for complementary vector control strategies, such as those involving lure-and-kill traps, or the development of mass trapping. This review also points out the necessity of further improving the search for new volatile attractants, such as new compound blends in specific ratios, considering that mosquito attraction to odors may vary over the life of the mosquito or among species. Finally, the use of mosquito attractants will undoubtedly have an increasingly important role to play in future integrated vector management programs.

Behavioural and antennal responses of Aedes aegypti (l.) (Diptera: Culicidae) gravid females to chemical cues from conspecific larvae.

Mass trapping of gravid females represents one promising strategy for the development of sustainable tools against Aedes aegypti. However, this technique requires the development of effective odorant lures that can compete with natural breeding sites. The presence of conspecific larvae has been shown to stimulate oviposition. Hence, we evaluated the role of four major molecules previously identified from Ae. aegypti larvae (isovaleric, myristoleic, myristic [i.e. tetradecanoic], and pentadecanoic acids) on the oviposition of conspecific females, as well as their olfactory perception to evaluate their range of detection. Using flight cage assays, the preference of gravid females to oviposit in water that previously contained larvae (LHW) or containing the four larval compounds was evaluated. Then, compounds and doses inducing the highest stimulation were challenged for their efficacy against LHW. Only isovaleric acid elicited antennal response, suggesting that the other compounds may act as taste cues. Pentadecanoic acid induced significant oviposition stimulation, especially when dosed at 10 ppm. Myristoleic acid and isovaleric acid deterred oviposition at 10 and 100 ppm, while no effect on oviposition was observed with myristic acid irrespectively of the dose tested. When the four compounds were pooled to mimic larvae's chemical signature, they favored oviposition at 1 ppm but negatively affected egg-laying at higher concentrations. When properly dosed, pentadecanoic acid and the blend of compounds may be promising lures for ovitraps as they could compete with LHW. Due to their low volatility, their effect should be further evaluated under field conditions, in addition with long-range attractants for developing effective tools against gravid females.

Prior contact with permethrin decreases its irritancy at the following exposure among a pyrethroid-resistant malaria vector Anopheles gambiae.

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.

Efficacy of vector control tools against malaria-infected mosquitoes.

Within mosquito vector populations, infectious mosquitoes are the ones completing the transmission of pathogens to susceptible hosts and they are, consequently, of great epidemiological interest. Mosquito infection by malaria parasites has been shown to affect several traits of mosquito physiology and behavior, and could interplay with the efficacy of control tools. In this study, we evaluated, in pyrethroid resistant Anopheles gambiae, the effect of mosquito infection with the human malaria parasite Plasmodium falciparum on the efficacy of nets treated with either the insecticide deltamethrin or the repellent DEET, measuring (i) mosquito success to pass through the net, (ii) blood-feeding on a host and (iii) chemicals-induced mortality. Infection of mosquitoes at non-infectious stage did not affect their success to pass through the net, to blood-feed, nor chemicals-induced mortality. At infectious stage, depending on replicates, infected mosquitoes had higher mortality rates than uninfected mosquitoes, with stronger effect in presence of DEET. This data evidenced a cost of infection on mosquito survival at transmissible stages of infection, which could have significant consequences for both malaria epidemiology and vector control. This stresses the need for understanding the combined effects of insecticide resistance and infection on the efficacy on control tools.

DEET Efficacy Increases With Age in the Vector Mosquitoes Anopheles gambiae s.s. and Aedes albopictus (Diptera: Culicidae).

Old mosquitoes are the most likely to transmit pathogens due to the higher probability that they will be exposed to pathogens, and the time required before a mosquito becomes infectious, the extrinsic incubation period (EIP). However, old mosquitoes are rarely considered in the evaluation of control tools. This study evaluated the effect of mosquito aging on the repellent efficacy of N,N-diethyl-3-methylbenzamide (DEET) in two vector mosquitoes, Anopheles gambiae s.s. (Giles) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae), respective vectors of malaria parasites and arboviruses such as dengue, chikungunya, and Zika viruses. For both mosquito species, DEET-induced inhibition of blood-feeding was assessed in three age classes. Life-history traits related to mosquito fecundity and survival following DEET exposure were also measured. Results showed that, in both species, bloodmeal inhibition induced by DEET was significantly higher in old females (>18 d old) than in younger ones (<13 d old). Life history traits recording showed no combined effects of DEET and aging on mosquito blood engorgement, oviposition rate, number of eggs laid nor survival; however, age effects are reported for all these traits. These results highlight the need for taking into account mosquito age in the evaluation of integrated mosquito management tools. They also suggest that the control of vector-borne pathogens with a long EIP could be improved by targeting old mosquitoes and supports the efficacy of repellents in the fight against mosquito-borne diseases.

Effect of DEET-multiple exposures on behavior and life history traits in the malaria mosquito Anopheles gambiae (s.s.).

BACKGROUND: Vector-borne diseases are major public health concerns, and their control is threatened by the spread of insecticide resistance in vector populations. In this context, the use of repellents is an alternative approach to limit vector-host interactions. However, prior exposure to repellents is suspected to affect mosquito behavior at the subsequent exposure, possibly reducing the efficacy of the compound. Despite this, the effect of mosquito experience on repellent efficacy remains poorly documented. In the present study, we tested whether a first blood meal successfully obtained upon a DEET-treated net would affect the success at taking a second blood meal in spite of DEET in the malaria mosquito Anopheles gambiae (s.s.). The impact of DEET on mosquito life history traits after the first and the second exposure was also measured, in order to assess the long-term consequences of multiple exposures to DEET in vector insects. RESULTS: A first blood meal obtained upon a DEET-treated net did not influence the success of An. gambiae females to take a second blood meal in spite of DEET. However, data showed that a prior exposure to DEET negatively affected all life history traits tested in this study related to fecundity and fertility. DEET pre-exposed females displayed a reduction in blood engorgement at the second exposure, as well as a reduction in the number of eggs laid and in the proportion of offspring that reach adult stage. Also, an increase of mosquito activity was observed during the second blood meal in DEET-pre-exposed females. Taken together, these data suggest an overall impact of DEET exposure on mosquito fitness. CONCLUSIONS: Our results did not evidence any effect of a prior exposure to DEET on its efficacy during the second exposure. However, data show a negative impact of DEET exposure on mosquito fitness. These results give insights to understand the long-term efficacy of the most used mosquito repellent, and highlight that DEET induces deleterious effects on mosquito fitness in addition to repellency, potentially increasing its efficacy for controlling vector-borne diseases.