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Invasive mosquito species are key vectors of arboviral diseases, like dengue, zika and chikungunya, posing significant public health challenges worldwide. These issues are worsened by urbanization, climate change and insecticide resistance, driving research into new control methods. Adult mosquitoes are attracted to plant nutrient sources essential for flight and reproduction. However, few studies have examined the odours emitted by these sources, and little is known about the olfactory neurobiology of mosquito–plant interactions. This review synthesizes current knowledge on the broad classes of volatile compounds that modulate mosquito behaviour, focusing on the olfactory processes underlying mosquito responses to plant nutrient sources. We also discuss the application of neurogenetic tools for investigating the role of olfactory receptor genes and neural circuits in mosquito ecology. Finally, we explore how insights from these studies can inform and enhance mosquito control strategies, including developing synthetic lures for attractive toxic sugar baits and improved trapping and surveillance technologies. Defining the olfactory receptors, sensory neurons and neural circuits mediating attraction or repellency to plant odours is crucial for optimizing mosquito monitoring and control interventions.more » « less
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The most devastating disease affecting the global citrus industry is Huanglongbing (HLB), caused by the pathogen Candidatus Liberibacter asiaticus. HLB is primarily spread by the insect vector Diaphorina citri (Asian Citrus Psyllid). To counteract the rapid spread of HLB by D. citri, traditional vector control strategies such as insecticide sprays, the release of natural predators, and mass introductions of natural parasitoids are used. However, these methods alone have not managed to contain the spread of disease. To further expand the available tools for D. citri control through generating specific modifications of the D. citri genome, we have developed protocols for CRISPR-Cas9-based genetic modification. Until now, genome editing in D. citri has been challenging due to the general fragility and size of D. citri eggs. Here we present optimized methods for collecting and preparing eggs to introduce the Cas9 ribonucleoprotein (RNP) into early embryos and alternative methods of injecting RNP into the hemocoel of adult females for ovarian transduction. Using these methods, we have generated visible somatic mutations, indicating their suitability for gene editing in D. citri. These methods represent the first steps toward advancing D. citri research in preparation for future genetic-based systems for controlling HLB.more » « less
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Abstract MultipleWolbachiastrains can block pathogen infection, replication and/or transmission inAedes aegyptimosquitoes under both laboratory and field conditions. However,Wolbachiaeffects on pathogens can be highly variable across systems and the factors governing this variability are not well understood. It is increasingly clear that the mosquito host is not a passive player in whichWolbachiagoverns pathogen transmission phenotypes; rather, the genetics of the host can significantly modulateWolbachia‐mediated pathogen blocking. Specifically, previous work linked variation inWolbachiapathogen blocking to polymorphisms in the mosquito alpha‐mannosidase‐2 (αMan2) gene. Here we use CRISPR‐Cas9 mutagenesis to functionally test this association. We developed αMan2 knockouts and examined effects on bothWolbachiaand virus levels, using dengue virus (DENV;Flaviviridae) and Mayaro virus (MAYV;Togaviridae).Wolbachiatitres were significantly elevated in αMan2 knockout (KO) mosquitoes, but there were complex interactions with virus infection and replication. InWolbachia‐uninfected mosquitoes, the αMan2 KO mutation was associated with decreased DENV titres, but in aWolbachia‐infected background, the αMan2 KO mutation significantly increased virus titres. In contrast, the αMan2 KO mutation significantly increased MAYV replication inWolbachia‐uninfected mosquitoes and did not affectWolbachia‐mediated virus blocking. These results demonstrate that αMan2 modulates arbovirus infection inA. aegyptimosquitoes in a pathogen‐ andWolbachia‐specific manner, and thatWolbachia‐mediated pathogen blocking is a complex phenotype dependent on the mosquito host genotype and the pathogen. These results have a significant impact for the design and use ofWolbachia‐based strategies to control vector‐borne pathogens.more » « less
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