Mosquito oviposition site selection is essential for vector population dynamics and malaria epidemiology. Irrigated rice cultivations provide ideal larval habitats for malaria mosquitoes, which has resulted in increased prevalence of the malaria vector, Anopheles arabiensis, in sub-Saharan Africa. The nature and origin of the cues regulating this behaviour are only now being elucidated. We show that gravid Anopheles arabiensis are attracted and oviposit in response to the odour present in the air surrounding rice. Furthermore, we identify a synthetic rice odour blend, using electrophysiological and chemical analyses, which elicits attraction and oviposition in laboratory assays, as well as attraction of free-flying gravid mosquitoes under semi-field conditions. This research highlights the intimate link between malaria vectors and agriculture. The identified volatile cues provide important substrates for the development of novel and cost-effective control measures that target female malaria mosquitoes, irrespective of indoor or outdoor feeding and resting patterns.

BackgroundAnopheles arabiensis is a key vector for the transmission of human malaria in sub-Saharan Africa. Over the past 10,000 years, humans have successfully cultivated grasses and altered the landscape, creating An. arabiensis favourable environments that contain excellent habitats for both larvae and adults. Sugarcane is the most expanding agricultural system in sub-Saharan Africa, and is linked to the increased threat of malaria in rural communities. The prolific production and wind dispersal of sugarcane pollen, together with standing pools of water, often provide, as a result of irrigation, a nutrient-rich environment for the offspring of gravid malaria mosquitoes.ResultsIn the present study, sugarcane pollen-associated volatiles from two cultivars are shown to attract gravid An. arabiensis in a still air two-port olfactometer and stimulate egg laying in an oviposition bioassay. Through combined gas chromatography and electroantennographic detection, as well as combined gas chromatography and mass spectrometric analyses, we identified the bioactive volatiles and generated a synthetic blend that reproduced the full behavioural repertoire of gravid mosquitoes in the Y-tube assay. Two subtractive odour blends, when compared with the full blend, were significantly more attractive. These three and four-component subtractive blends share the compounds (1R)-(+)-α-pinene, nonanal and benzaldehyde, of which, (1R)-(+)-α-pinene and nonanal are found in the attractive odour blends from rice plants and maize pollen. In pairwise comparisons, the rice synthetic odour blend was more attractive to gravid mosquitoes than either of the pollen blends, whereas the pollen blends did not differ in attraction.ConclusionsThe attraction of gravid females to sugarcane pollen volatiles demonstrated in this study, together with the previously found grass-associated volatiles, raise the potential of developing a bioactive chimeric blend to attract gravid malaria mosquitoes. This is discussed in relation to the development of novel and cost-effective vector control measures.

BackgroundMaize cultivation contributes to the prevalence of malaria mosquitoes and exacerbates malaria transmission in sub-Saharan Africa. The pollen from maize serves as an important larval food source for Anopheles mosquitoes, and females that are able to detect breeding sites where maize pollen is abundant may provide their offspring with selective advantages. Anopheles mosquitoes are hypothesized to locate, discriminate among, and select such sites using olfactory cues, and that synthetic volatile blends can mimic these olfactory-guided behaviours.MethodsTwo-port olfactometer and two-choice oviposition assays were used to assess the attraction and oviposition preference of gravid Anopheles arabiensis to the headspace of the pollen from two maize cultivars (BH-660 and ZM-521). Bioactive compounds were identified using combined gas chromatography and electroantennographic detection from the headspace of the cultivar found to be most attractive (BH-660). Synthetic blends of the volatile compounds were then assessed for attraction and oviposition preference of gravid An. arabiensis, as above.ResultsHere the collected headspace volatiles from the pollen of two maize cultivars was shown to differentially attract and stimulate oviposition in gravid An. arabiensis. Furthermore, a five-component synthetic maize pollen odour blend was identified, which elicited the full oviposition behavioural repertoire of the gravid mosquitoes.ConclusionsThe cues identified from maize pollen provide important substrates for the development of novel control measures that modulate gravid female behaviour. Such measures are irrespective of indoor or outdoor feeding and resting patterns, thus providing a much-needed addition to the arsenal of tools that currently target indoor biting mosquitoes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-016-1656-0) contains supplementary material, which is available to authorized users.

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Background Odour-based tools targeting gravid malaria vectors may complement existing intervention strategies. Anopheles arabiensis are attracted to, and stimulated to oviposit by, natural and synthetic odours of wild and domesticated grasses associated with mosquito breeding sites. While such synthetic odour lures may be used for vector control, these may have limited efficacy when placed in direct competition with the natural source. In this study, workflows developed for plant-feeding pests was used to design and evaluate a chimeric odour blend based on shared attractive compounds found in domesticated grass odours. Methods Variants of a synthetic odour blend, composed of shared bioactive compounds previously identified in domesticated grasses, was evaluated sequentially in a two-choice olfactometer to identify a ratio-optimized attractive blend for malaria vectors. During this process, blends with ratios that were significantly more attractive than the previously identified synthetic rice blend were compared to determine which was most attractive in the two-choice olfactometer. To determine whether all volatile components of the most attractive blend were necessary for maximal attraction, subtractive assays were then conducted, in which individual components were removed for the most attractive blend, to define the final composition of the chimeric blend. Binary logistic regression models were used to determine significance in all two-choice assays. The chimeric blend was then assessed under field conditions in malaria endemic villages in Ethiopia, to assess the effect of dose, trap type, and placement relative to ground level. Field data were analyzed both descriptively and using a Welch-corrected t-test. Results A ratio-optimized chimeric blend was identified that significantly attracted gravid An. arabiensis under laboratory conditions. In the field, trap captures of An. arabiensis and Anopheles pharoensis were dependent on the presence of the lure, trap type (CDC, BG Sentinel and Suna traps), placement relevant to ground level, with low release rates generally luring more mosquitoes. Conclusions The workflow designed for the development of chimeric lures provides an innovative strategy to target odour-mediated behaviours. The chimeric lure identified here can be used in existing trapping systems, and be customized to increase sustainability, in line with goals of the Global Vector Control Response Group.

Collecting blood‐fed mosquitoes to monitor pathogen presence or to gather information on the host blood meal is often challenging. Fermenting molasses can be used to produce carbon dioxide to attract host‐seeking mosquitoes, however, earlier work indicated that it may also attract blood‐fed mosquitoes in the field. In the current study, these field results were validated in an experimental setting using a large cage setup with Anopheles coluzzii (Diptera, Culicidae). Blood‐fed mosquitoes were indeed attracted to fermenting molasses with the highest attraction at 72 hours post feeding, which was used for subsequent experiments. Next, it was tested if fermentation of molasses is required for attraction, and whether it acts as an oviposition attractant, increases egg laying, or increases mosquito survival. The compounds that could be responsible for attraction were identified by combined electrophysiology and chemical analyses and formulated into a synthetic blend. Fermenting molasses attracted blood‐fed mosquitoes in the large cage study, while fermenting sugar and non‐fermenting molasses did not. The fecundity of blood‐fed mosquitoes increased after feeding on fermenting molasses, however, compounds emanating from molasses did not trigger oviposition. The synthetic blend attracted blood‐fed mosquitoes and may be used to determine mosquito host selection and for xenomonitoring, as ‘flying syringes’ to detect non‐vector borne pathogens.

Mosquitoes use chemical codes to locate and discriminate among vertebrate hosts to obtain a blood meal. Recent advances have allowed for the identification of the chemical codes used by mosquitoes to locate and discriminate humans from other vertebrate hosts. Humans are incidental “dead-end” hosts for the West Nile virus, which is maintained in an enzootic cycle, primarily through its transmission between infected birds by Culex mosquitoes. Host-seeking Culex mosquitoes are attracted to the odor of chicken, which are used in sentinel traps to monitor West Nile virus transmission. Using combined gas chromatography and electroantennography and mass spectrometry we identify a blend of volatile organic compounds present in chicken emanates, including mostly salient bioactive compounds previously identified in human emanates. When released at their identified ratios, this blend elicits behavioral responses of Culex pipiens molestus and Culex quinquefasciatus similar to that to the natural chicken odor. Tested under field conditions, this blend attract Culex spp. and other species of mosquitoes using birds among their hosts. This study provides evidence for conserved chemical codes for resource location by mosquitoes, and highlights the intricate role of CO2 for host-seeking mosquitoes. The identification of conserved chemical codes, which drive innate preference behaviors that are fundamental for survival and reproduction, provides important substrates for future control interventions targeting disease vector mosquitoes.

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