The mosquito Aedes aegypti is the primary vector for the fastest growing infectious disease in the world, dengue fever. Disease transmission heavily relies on the ability of female mosquitoes to locate their human hosts. Additionally, males may be found in close proximity to humans, where they can find mates. Host seeking behaviour of both sexes is dependent on adult sexual maturation. Identifying the molecular basis for the onset of host seeking may help to determine targets for future vector control. In this study, we investigate modulation of the host seeking behaviour and the transcript abundance of the main chemoreceptor families between sexes and across ages in newly-emerged mosquitoes. Attraction to human odour was assessed using a Y-tube olfactometer, demonstrating that both males and females display age-dependent regulation of host seeking. The largest increase in transcript abundance was identified for select chemosensory genes in the antennae of young adult Ae. aegypti mosquitoes and reflects the increase in attraction to human odour observed between 1 and 3 day(s) post-emergence in both males and females. Future functional characterisation of the identified differentially abundant genes may provide targets for the development of novel control strategies against vector borne diseases.

Pathogens may manipulate their human and mosquito hosts to enhance disease transmission. Dengue, caused by four viral serotypes, is the fastest-growing transmissible disease globally resulting in 50-100 million infections annually. Transmission of the disease relies on the interaction between humans and the vector Aedes aegypti and is largely dependent on the odor-mediated host seeking of female mosquitoes. In this study, we use activity monitors to demonstrate that dengue virus-1 affects the locomotion and odor-mediated behavior of Ae. aegypti, reflecting the progression of infection within the mosquito. Mosquitoes 4-6 days post-infection increase locomotion, but do not alter their odor-driven host-seeking response. In contrast, females 14-16 days post-infection are less active, yet more sensitive to human odors as assessed by behavioral and electrophysiological assays. Such an increase in physiological and behavioral sensitivity is reflected by the antennal-specific increase in abundance of neural signaling transcripts in 14 days post-infection females, as determined by transcriptome analysis. This suggests that the sensitivity of the mosquito peripheral olfactory system is altered by the dengue virus by enhancing the overall neural responsiveness of the antenna, rather than the selective regulation of chemosensoryrelated genes. Our study reveals that dengue virus-1 enhances vector-related behaviors in the early stages post-infection that aid in avoiding predation and increasing spatial exploration. On the other hand, at the later stages of infection, the virus enhances the host-seeking capacity of the vector, thereby increasing the risk of virus transmission. A potential mechanism is discussed.

The Queensland fruit fly (Bactrocera tryoni) is a major polyphagous pest widespread in Australia and several Pacific Islands. Bacteria present on the host plant phyllosphere supply proteins, essential for egg development and female sexual maturity. We investigated the role of microbial volatile organic compounds (MVOCs) in attracting virgin females. Four Enterobacteriaceae commonly found on the host plant and in the gut of B. tryoni were cultured on different media (artificial and natural fruits), and their MVOCs were collected using two different headspace volatile absorbent materials (Tenax® filter and solid phase micro-extraction fibre; SPME), allowing to test for time and pH variations. The olfactory responses of virgin females to bacterial MVOCs were assessed via electrophysiology and behavioural assays. The production of MVOCs was strongly influenced qualitatively by the bacterial strain, the type of media and quantitatively varied with pH and time. Phenol was detected only from Citrobacter freundii (CF), while indole was found in all bacteria except CF. Indole production increased overtime in inoculated kiwifruits. D-limonene was detected from Enterobacter cloacae and Klebsiella oxytoca (KO) and associated with oranges and inoculated kiwifruit. Tenax extracts from KO invoked the strongest antennal response and attracted virgin females the most. Among the identified compounds triggering an olfactory response, D-limonene and 2-nonanone were both significantly behaviourally attractive whereas phenol, nonanal, isoamyl alcohol and some pyrazines appeared repulsive. This study provides a new foundation for developing a synthetic lure based on microbial volatiles to replace food baits targeting specifically virgin females.

The Queensland fruit fly (Bactrocera tryoni) is a major polyphagous pest widespread in Australia and several Pacific Islands. Bacteria present on the host plant phyllosphere supply proteins, essential for egg development and female sexual maturity. We investigated the role of microbial volatile organic compounds (MVOCs) emitted by Enterobacteriaceae commonly found on the host plant and in the fly gut in attracting virgin females. Bacteria were cultured on artificial media and natural fruits, at various pH, and MVOCs were collected using different headspace volatile absorbent materials. The olfactory responses of virgin females to bacterial MVOCs were assessed via electrophysiology and behavioral assays. The production of MVOCs was strongly influenced qualitatively by the bacterial strain and the type of media, and it semi-quantitatively varied with pH and time. MVOCs emitted by Klebsiella oxytoca invoked the strongest antennal response and were the most attractive. Among the identified compounds triggering an olfactory response, D-limonene and 2-nonanone were both significantly behaviorally attractive, whereas phenol, nonanal, isoamyl alcohol, and some pyrazines appeared to be repulsive. This study deepens our understanding of the chemical ecology between fruit flies and their bacterial symbionts and paves the way for novel synthetic lures based on specifically MVOCs targeting virgin females.