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Behavioral responses of Aedes aegypti females to the different in vitro community models and emitted volatile compounds by the different community models, in competitive environments. (A) Number of landings in the arena by Ae. aegypti. (B) Time spent in the arena per individual (s). (C) Dendrogram and heatmap of the different volatiles emitted by the community models with increased complexity. (D) Dendrogram and heatmap of the different volatiles emitted by the community models reflecting the skin bacterial composition of highly and poorly attractive individuals. Volatile organic compounds appear in the order of elution. Blue shades should be read as traces or absent. Staphylococcus epidermidis was the reference control grown as a single model. Double model was composed of S. epidermidis, Pseudomonas aeruginosa in a 1:1 ratio; Triple was composed of S. epidermidis, P. aeruginosa, Corynebacterium minutissimum in a 1:1:1 ratio; Quadruple was composed of S. epidermidis, P. aeruginosa, C. minutissimum, Brevibacterium epidermidis in a 1:1:1:1 ratio; Highly Attractive (HA) community was composed of S. epidermidis, P. aeruginosa, C. minutissimum, B. epidermidis in a 3:3:2:8 ratio and the Poorly Attractive community was composed of S. epidermidis, P. aeruginosa, C. minutissimum, B. epidermidis in a 1:6:1:8 ratio. Behavioral data was analyzed by general linear model with a negative binomial distribution, using likelihood function and chi-square test (A) or by generalized linear model with gamma distribution using likelihood function and chi-square test (B). Tukey’s post-hoc test was used for pairwise comparisons at the 0.05 significance level. Lowercase letters indicate significant differences between treatments at the 0.05 level. Outliers are represented by circles (out) and stars (far out). Dendrogram clustering was performed using Ward’s clustering algorithm with Euclidean distances (C,D). We had 8 biological replicates per treatment.
Original Research
25 May 2023
Competition matters: using in vitro community models to study the impact of human skin bacteria on mosquito attraction
Dani Lucas-Barbosa
6 more and 
Niels O. Verhulst

The human skin bacteria play an important role in the production of volatiles that attract mosquitoes. Using some of the most abundant human skin bacterial species, we created in vitro community models to assess whether increased microbial biodiversity could reduce human attractiveness to females of the dengue fever mosquito Aedes aegypti and whether co-culturing bacterial commensals affects overall attraction. More complex bacterial models were less attractive to female mosquitoes than the simplest models. For instance, the triple bacterial community model was approximately three times less attractive than Staphylococcus epidermidis alone. Our data show, for instance, that an in vitro community model mimicking the skin composition of a highly attractive individual to the anthropophilic Anopheles gambiae was also more attractive to anthropophilic Ae. aegypti than a community model mimicking the skin composition of a poorly attractive individual to An. gambiae. In line with these results, volatile analyses of the blends emitted by the different in vitro community models showed that the more complex models had lower emission overall. Effects on mosquito responses differed sharply when the different bacteria species were sharing the same resources used for growth, showing that either competition or commensalism may influence their relative growth, and that this consequently can influence mosquito responses. We conclude that studies on mosquito responses to skin volatiles need to take the microbial community into account.

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Microbes communicate with each other using a wide array of chemical compounds, including volatile organic compounds (VOCs). Usually, such volatile-mediated interactions are studied by growing two different microbes in a shared, confined environment and by subsequently collecting and analyzing the emitted VOCs by gas chromatography. This procedure has several drawbacks, including artificial volatile overaccumulation and potential oxygen limitation, as well as the impossibility to assign a producer to the compounds newly emitted during the interaction. To address these challenges, we have developed a novel system specifically designed to analyze volatile-mediated interactions allowing for sequential unidirectional exposure of a “receiver” microorganism to the VOCs of an “emitter” microorganism. Using hermetically sealed systems connected to an air compressor, a constant unidirectional airflow could be generated, driving emitted volatiles to be absorbed by a collection charcoal filter. Thus, our developed system avoids artificial overaccumulation of volatile compounds and lack of oxygen in the headspace and enables the univocal assignment of VOCs to their producers. As a proof of concept, we used this newly developed experimental setup to characterize the reaction of plant growth-promoting and biocontrol fungus (Trichoderma simmonsii) to the perception of VOCs emitted by two plant pathogens, namely Botrytis cinerea and Fusarium oxysporum. Our results show that the perception of each pathogen's volatilome triggered a specific response, resulting in significant changes in the VOCs emitted by Trichoderma. Trichoderma's volatilome modulation was overall stronger when exposed to the VOCs from Fusarium than to the VOCs from Botrytis, which correlated with increased siderophore production when co-incubated with this fungus. Our newly developed method will not only help to better understand volatile-mediated interactions in microbes but also to identify new molecules of interest that are induced by VOC exposure, as well as the putative-inducing signals themselves.

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5 citations
3,743 views
4 citations
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Frontiers in Ecology and Evolution

Research Advances on Drosophila suzukii - Volume II
Edited by John Abraham, Sergio Angeli
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17 June 2025
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