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.

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.

Vector competence of Aedes aegypti from Havana, Cuba, for dengue virus type 1, chikungunya, and Zika viruses.

BACKGROUND: Like many countries from the Americas, Cuba is threatened by Aedes aegypti-associated arboviruses such as dengue (DENV), Zika (ZIKV), and chikungunya (CHIKV) viruses. Curiously, when CHIKV was actively circulating in the region in 2013-2014, no autochthonous transmission of this virus was detected in Havana, Cuba, despite the importation of chikungunya cases into this city. To investigate if the transmission ability of local mosquito populations could explain this epidemiological scenario, we evaluated for the first time the vector competence of two Ae. aegypti populations (Pasteur and Párraga) collected from Havana for dengue virus type 1 (DENV-1), CHIKV, and ZIKV. METHODOLOGY/PRINCIPAL FINDINGS: Mosquito populations were fed separately using blood containing ZIKV, DENV-1, or CHIKV. Infection, dissemination, and transmission rates, were estimated at 3 (exclusively for CHIKV), 7, and 14 days post exposure (dpe) for each Ae. aegypti population-virus combination. Both mosquito populations were susceptible to DENV-1 and ZIKV, with viral infection and dissemination rates ranging from 24-97% and 6-67% respectively. In addition, CHIKV disseminated in both populations and was subsequently transmitted. Transmission rates were low (<30%) regardless of the mosquito population/virus combination and no ZIKV was detected in saliva of females from the Pasteur population at any dpe. CONCLUSIONS/SIGNIFICANCE: Our study demonstrated the ability of Ae. aegypti from Cuba to transmit DENV, ZIKV, and CHIKV. These results, along with the widespread distribution and high abundance of this species in the urban settings throughout the island, highlight the importance of Ae. aegypti control and arbovirus surveillance to prevent future outbreaks.

Experimental Assessment of Zika Virus Mechanical Transmission by Aedes Aegypti.

The pandemic emergence of several mosquito-borne viruses highlights the need to understand the different ways in which they can be transmitted by vectors to human hosts. In this study, we evaluated the propensity of Aedes aegypti to transmit mechanically Zika virus (ZIKV) using an experimental design. Mosquitoes were allowed to feed on ZIKV-infected blood and were then rapidly transferred to feed on ZIKV-free blood until they finished their meal. The uninfected blood meals, the mosquito abdomens, as well as the mouthparts dissected from fully and partially engorged mosquitoes were analyzed using RT-qPCR and/or virus titration. All the fully engorged mosquito abdomens were ZIKV-infected, whereas their mouthparts were all ZIKV-negative. Nonetheless, one of the partially engorged mosquitoes carried infectious particles on mouthparts. No infectious virus was found in the receiver blood meals, while viral RNA was detected in 9% of the samples (2/22). Thus, mechanical transmission of ZIKV may sporadically occur via Ae. aegypti bite. However, as the number of virions detected on mouthparts (2 particles) is not sufficient to induce infection in a naïve host, our results indicate that mechanical transmission does not impact ZIKV epidemiology.

Transmission potential of African, Asian and American Zika virus strains by Aedes aegypti and Culex quinquefasciatus from Guadeloupe (French West Indies).

Zika virus (ZIKV) is an arbovirus that has dramatically spread in South America and the Caribbean regions since 2015. The majority of vector incrimination studies available for ZIKV showed that Aedes aegypti mosquitoes are important vectors for this virus. However, several reports suggest that Culex quinquefasciatus mosquitoes may be implicated in ZIKV transmission in certain urban settings. In the present study, we evaluated the vector competence for ZIKV of Cx. quinquefasciatus and Ae. aegypti mosquitoes from Guadeloupe using African, American and Asian strains. The results demonstrated that Cx. quinquefasciatus is refractory to ZIKV infection whatever the strain tested at 7, 14 or 21 days post-infection (dpi), while ZIKV transmission was recorded in Ae. aegypti for all the three strains. The African ZIKV strain was better transmitted by Ae. aegypti (∼ 50% mean transmission efficiency) and with a shorter incubation period (7 dpi) when compared to the Asian and American strains (<14% transmission efficiency; incubation period of 14-21 dpi). Taken together, these results suggest that only Ae. aegypti mosquitoes are involved in urban ZIKV transmission in Guadeloupe and highlight a higher infectiousness of the African ZIKV strain in this mosquito species when compared to the Asian and American ones.

Improving the Monitoring of the Walnut Husk Fly (Diptera: Tephritidae) Using Male-Produced Lactones.

It is important to monitor fruit flies (Diptera: Tephritidae) efficiently to implement sustainable means of control. Attractants are often used to increase the efficiency of sticky traps deployed in orchards to monitor Lepidopterans, but remains to be developed to monitor fruit flies. Rhagoletis completa Cresson (Diptera: Tephritidae) is an invasive species in the walnut orchards of Europe, and is commonly monitored with yellow sticky traps. In this study, we collected the volatile compounds released by male and female R. completa, and identified two lactones released exclusively by males. We then formulated both lactones in long-lasting volatile dispensers, and we quantified their release rate over a 26-d period. Finally, during the entire period when female flies are present in the field, we compared the efficiency of the conventional monitoring method using unbaited yellow sticky traps with yellow sticky traps associated with a dispenser releasing both male-produced lactones. These assays were conducted in 54 walnut orchards in France, in 2017. The number of fruit flies caught with sticky traps associated with lactones dispensers was increased by up to 10 times each week. Lactone-baited traps also allowed earlier detection in the season. These field results are promising for R. completa monitoring. A complete chiral identification of these lactones should be performed along with a clarification of their role in the sexual communication of R. completa.

Elevated CO2 Concentrations Impact the Semiochemistry of Aphid Honeydew without Having a Cascade Effect on an Aphid Predator.

Honeydew is considered a cornerstone of the interactions between aphids and their natural enemies. Bacteria activity occurring in aphid honeydew typically results in the release of volatile organic compounds (VOCs) that are used by the natural enemies of aphids to locate their prey. Because atmospheric carbon dioxide (CO2) concentration directly impacts the physiology of plants, we raise the hypothesis that elevated CO2 concentrations impact the quantity of honeydew produced by aphids, as well as the diversity and quantity of honeydew VOCs, leading to cascade effects on the foraging behavior of aphids' natural enemies. Using solid-phase microextraction, we analyzed the VOCs emitted by honeydew from pea aphids (Acyrthosiphon pisum Harris) reared under 450 ± 50 ppm of CO2 (aCO2) or 800 ± 50 ppm CO2 (eCO2). While the total amount of honeydew excreted was only slightly reduced by eCO2 concentrations, we detected qualitative and quantitative differences in the semiochemistry of aphid honeydew between CO2 conditions. Three VOCs were not found in the honeydew of eCO2 aphids: 3-methyl-2-buten-1-ol, 2-methyl-1-butanol, and isobutanol. However, no difference was observed in the searching and oviposition behaviors of hoverfly (Episyrphus balteatus (De Geer)) females exposed to plants covered with honeydew originating from the different CO2 conditions. The present work showed the effect of a particular aspect of atmospheric changes, and should be extended to other abiotic parameters, such as temperature.

Identification of the Alarm Pheromone of Cowpea Aphid, and Comparison With Two Other Aphididae Species.

In response to a predator attack, many Aphidinae species release an alarm pheromone, which induces dispersal behavior in other individuals within the colony. The major component of this pheromone is the sesquiterpene (E)-ß-farnesene (Eßf), but variations occur between aphid species. In the present work, we collected, identified, and quantified the alarm pheromone of Aphis craccivora Koch (Hemiptera: Aphididae), before quantifying the escape behavior induced in the neighboring individuals. We compared the semiochemistry and associated behavior of alarm signaling with two other aphid species: Myzus persicae (Sulzer) (Hemiptera: Aphididae) and Aphis fabae Scopoli (Hemiptera: Aphididae). Eßf was the only volatile found for each species. M. persicae produces a higher quantity of Eßf (8.39 ± 1.19 ng per individual) than A. craccivora (6.02 ± 0.82 ng per individual) and A. fabae (2.04 ± 0.33 ng per individual). Following exposure to natural doses of synthetic Eßf (50 ng and 500 ng), A. craccivora respond more strongly than the two other Aphidinae species with 78% of the individuals initiated alarm behavior for 500 ng of Eßf.

Oviposition deterrent activity of basil plants and their essentials oils against Tuta absoluta (Lepidoptera: Gelechiidae).

The leafminer Tuta absoluta Meyrick (Lepidoptera: Gelechiidae) is one of the most important pests of tomato, reducing crop yields by up to 100% in greenhouses and fields, in several countries globally. Because synthetic insecticides lead to resistance and have adverse effects on natural enemies and the health of producers, alternative control methods are needed. In this study, we assessed the oviposition-deterring effect of basil plants, Ocimum gratissimum L. and O. basilicum L. (Lamiaceae), using dual-choice behavioural assays performed in flight tunnels. We found that both plants significantly reduced T. absoluta oviposition behaviour on a tomato plant located nearby. To evaluate the potential effect of basil volatile organic compounds, we formulated essential oils of both plant species in paraffin oil, and observed a similar oviposition-deterring effect. Gas chromatography analyses detected 18 constituents in these essential oils which the major constituents included thymol (33.3%), p-cymene (20.4%), γ-terpinene (16.9%), myrcene (3.9%) in O. gratissimum and estragol (73.8%), linalool (8.6%), ß-elemene (2.9%) and E-ß-ocimene (2.6%) in O. basilicum. Twenty and 33 compounds were identified of the volatiles collected on O. gratissimum and O. basilicum plants, respectively. The main components include the following: p-cymene (33.5%), γ-terpinene (23.6%), α-terpinene (7.2%), α-thujene (6.7%) and E-α-bergamotene (38.9%) in O. gratissimum, and methyl eugenol (26.1%), E-ß-ocimene (17.7%), and linalool (9.4%) in O. basilicum. Four compounds (α-pinene, ß-pinene, Myrcene, Limonene) were common in essential oils and plants. Our results suggest the valuable potential of basil and associated essential oils as a component of integrated management strategies against the tomato leafminer.

First Characterisation of Volatile Organic Compounds Emitted by Banana Plants.

Banana (Musa sp.) ranks fourth in term of worldwide fruit production, and has economical and nutritional key values. The Cavendish cultivars correspond to more than 90% of the production of dessert banana while cooking cultivars are widely consumed locally around the banana belt production area. Many plants, if not all, produce Volatile Organic Compounds (VOCs) as a means of communication with their environment. Although flower and fruit VOCs have been studied for banana, the VOCs produced by the plant have never been identified despite their importance in plant health and development. A volatile collection methodology was optimized to improve the sensitivity and reproducibility of VOCs analysis from banana plants. We have identified 11 VOCs for the Cavendish, mainly (E,E)-α-farnesene (87.90 ± 11.28 ng/µl), methyl salicylate (33.82 ± 14.29) and 6-methyl-5-hepten-2-one (29.60 ± 11.66), and 14 VOCs for the Pacific Plantain cultivar, mainly (Z,E)-α-farnesene (799.64 ± 503.15), (E,E)-α-farnesene (571.24 ± 381.70) and (E) ß ocimene (241.76 ± 158.49). This exploratory study paves the way for an in-depth characterisation of VOCs emitted by Musa plants.

Identification of walnut husk (Juglans regia L.) volatiles and the behavioural response of the invasive Walnut Husk Fly, Rhagoletis completa Cresson.

BACKGROUND: Several European countries are important walnut (Juglans regia L.) producers. However, these countries must contend with the recent introduction of the Walnut Husk Fly, Rhagoletis completa Cresson (Diptera, Tephritidae), which is causing severe economic losses, especially in organic production. Because most Tephritid fruit flies use kairomones in their search for host plants, we hypothesise that this highly specialist species orients toward the volatile blend released by walnut husks. RESULTS: We collected, identified, and quantified the volatile organic chemicals (VOCs) released by walnut husks from the most commonly cultivated variety in France (Franquette). Then, the behavioural response of R. completa toward synthetic odour blends was recorded in dual choice assays conducted in net cages. A total of 26 VOCs were identified, with α-pinene, ß-pinene, trans-linalool, eugenol, and tetradecane representing the major constituents. In the dual choice assay, male and female R. completa were strongly attracted to synthetic blend that included most of the identified husk VOCs. CONCLUSION: When searching for a host plant, R. completa use host fruit kairomones. The potential of these semiochemicals in monitoring and management of this quarantine pest is discussed. © 2017 Society of Chemical Industry.

Elevated Carbon Dioxide Concentration Reduces Alarm Signaling in Aphids.

Insects often rely on olfaction to communicate with conspecifics. While the chemical language of insects has been deciphered in recent decades, few studies have assessed how changes in atmospheric greenhouse gas concentrations might impact pheromonal communication in insects. Here, we hypothesize that changes in the concentration of atmospheric carbon dioxide affect the whole dynamics of alarm signaling in aphids, including: (1) the production of the active compound (E)-ß-farnesene (Eßf), (2) emission behavior when under attack, (3) perception by the olfactory apparatus, and (4) the escape response. We reared two strains of the pea aphid, Acyrthosiphon pisum, under ambient and elevated CO2 concentrations over several generations. We found that an increase in CO2 concentration reduced the production (i.e., individual content) and emission (released under predation events) of Eßf. While no difference in Eßf neuronal perception was observed, we found that an increase in CO2 strongly reduced the escape behavior expressed by an aphid colony following exposure to natural doses of alarm pheromone. In conclusion, our results confirm that changes to greenhouse gases impact chemical communication in the pea aphid, and could potentially have a cascade effect on interactions with higher trophic levels.

Will climate change affect insect pheromonal communication?

Understanding how climate change will affect species interactions is a challenge for all branches of ecology. We have only limited understanding of how increasing temperature and atmospheric CO2 and O3 levels will affect pheromone-mediated communication among insects. Based on the existing literature, we suggest that the entire process of pheromonal communication, from production to behavioural response, is likely to be impacted by increases in temperature and modifications to atmospheric CO2 and O3 levels. We argue that insect species relying on long-range chemical signals will be most impacted, because these signals will likely suffer from longer exposure to oxidative gases during dispersal. We provide future directions for research programmes investigating the consequences of climate change on insect pheromonal communication.

Climate Change and Tritrophic Interactions: Will Modifications to Greenhouse Gas Emissions Increase the Vulnerability of Herbivorous Insects to Natural Enemies?

Insects are highly dependent on odor cues released into the environment to locate conspecifics or food sources. This mechanism is particularly important for insect predators that rely on kairomones released by their prey to detect them. In the context of climate change and, more specifically, modifications in the gas composition of the atmosphere, chemical communication-mediating interactions between phytophagous insect pests, their host plants, and their natural enemies is likely to be impacted. Several reports have indicated that modifications to plants caused by elevated carbon dioxide and ozone concentrations might indirectly affect insect herbivores, with community-level modifications to this group potentially having an indirect influence on higher trophic levels. The vulnerability of agricultural insect pests toward their natural enemies under elevated greenhouse gases concentrations has been frequently reported, but conflicting results have been obtained. This literature review shows that the higher levels of carbon dioxide, as predicted for the coming century, do not enhance the abundance or efficiency of natural enemies to locate hosts or prey in most published studies. Increased ozone levels lead to modifications in herbivore-induced volatile organic compounds (VOCs) released by damaged plants, which may impact the attractiveness of these herbivores to the third trophic level. Furthermore, other oxidative gases (such as SO2 and NO2) tend to reduce the abundance of natural enemies. The impact of changes in atmospheric gas emissions on plant-insect and insect-insect chemical communication has been under-documented, despite the significance of these mechanisms in tritrophic interactions. We conclude by suggesting some further prospects on this topic of research yet to be investigated.