BackgroundMosquitoes 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.MethodsExperiments 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.ResultsWe 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.ConclusionsThese 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 materialThe online version of this article (doi:10.1186/s13071-016-1634-y) contains supplementary material, which is available to authorized users.
BackgroundInsect repellents are prophylactic tools against a number of vector-borne
diseases. There is growing demand for repellents outperforming DEET in cost
and safety, but with the current technologies R&D of a new product takes
almost 10 years, with a prohibitive cost of $30 million dollar in
part due to the demand for large-scale synthesis of thousands of test
compounds of which only 1 may reach the market. R&D could be expedited
and cost dramatically reduced with a molecular/physiological target to
streamline putative repellents for final efficacy and toxicological
tests.MethodologyUsing olfactory-based choice assay we show here that the fruit fly is
repelled by not only DEET, but also IR3535 and picaridin thus suggesting
they might have “generic repellent detector(s),” which may be of
practical applications in new repellent screenings. We performed single unit
recordings from all olfactory sensilla in the antennae and maxillary palps.
Although the ab3A neuron in the wild type flies responded to picaridin, it
was unresponsive to DEET and IR3535. By contrast, a neuron housed in the
palp basiconic sensilla pb1 responded to DEET, IR3535, and picaridin, with
apparent sensitivity higher than that of the DEET detectors in the
mosquitoes Culex quinquefasciatus and Aedes
aegypti. DmOr42a was transplanted from pb1 to the “empty
neuron” and showed to be sensitive to the three insect repellents.ConclusionsFor the first time we have demonstrated that the fruit fly avoids not only
DEET but also IR3535 and picaridin, and identified an olfactory receptor
neuron (ORN), which is sensitive to these three major insect repellents. We
have also identified the insect repellent-sensitive receptor, DmOr42a. This
generic detector fulfils the requirements for a simplified bioassay for
early screening of test insect repellents.
It is suggested that the different mechanisms of action of the larvicidal actives A. muricata acetogenins and P. nigrum piperamides explain the observed synergism. The combination of inexpensive botanicals and a low-cost organosolvent such as ethanol leads to a simple and efficient phytolarvicidal formulation.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.