Genetic control methods of mosquito vectors of malaria, dengue, yellow fever, and Zika are becoming increasingly popular due to the limitations of other techniques such as the use of insecticides. The sterile insect technique is an effective genetic control method to manage insect populations. However, it is crucial to release sterile mosquitoes by air to ensure homogeneous coverage, especially in large areas. Here, we report a fully automated adult mosquito release system operated from an uncrewed aerial vehicle or drone. Our system, developed and tested in Brazil, enabled a homogeneous dispersal of sterile male Aedes aegypti while maintaining their quality, leading to a homogeneous sterile-to-wild male ratio due to their aggregation in the same sites. Our results indicate that the released sterile males were able to compete with the wild males in mating with the wild females; thus, the sterile males were able to induce sterility in the native female population. The use of drones to implement the sterile insect technique will lead to improvements in areal coverage and savings in operational costs due to the requirement of fewer release sites and field staff.

Aedes aegypti is the primary vector of arthropod-borne viruses including dengue, chikungunya and Zika. Vector population control methods are reviving to impede disease transmission. An efficient sex separation for male-only releases is crucial for area-wide mosquito population suppression strategies. Here, we report on the construction of two genetic sexing strains using red- and white-eye colour mutations as selectable markers. Quality control analysis showed that the Red-eye genetic sexing strains (GSS) is better and more genetically stable than the White-eye GSS. The introduction of an irradiation-induced inversion (Inv35) increases genetic stability and reduces the probability of female contamination of the male release batches. Bi-weekly releases of irradiated males of both the Red-eye GSS and the Red-eye GSS/Inv35 fully suppressed target laboratory cage populations within six and nine weeks, respectively. An image analysis algorithm allowing sex determination based on eye colour identification at the pupal stage was developed. The next step is to automate the Red-eye-based genetic sexing and validate it in pilot trials prior to its integration in large-scale population suppression programmes. This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.

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BackgroundSeveral mosquito population suppression strategies based on the rearing and release of sterile males have provided promising results. However, the lack of an efficient male selection method has hampered the expansion of these approaches into large-scale operational programmes. Currently, most of these programmes targeting Aedes mosquitoes rely on sorting methods based on the sexual size dimorphism (SSD) at the pupal stage. The currently available sorting methods have not been developed based on biometric analysis, and there is therefore potential for improvement. We applied an automated pupal size estimator developed by Grupo Tragsa with laboratory samples of Anopheles arabiensis, Aedes albopictus, Ae. polynesiensis, and three strains of Ae. aegypti. The frequency distribution of the pupal size was analyzed. We propose a general model for the analysis of the frequency distribution of mosquito pupae in the context of SSD-sorting methods, which is based on a Gaussian mixture distribution functions, thus making possible the analysis of performance (% males recovery) and purity (% males on the sorted sample).ResultsFor the three Aedes species, the distribution of the pupae size can be modeled by a mixture of two Gaussian distribution functions and the proposed model fitted the experimental data. For a given population, each size threshold is linked to a specific outcome of male recovery. Two dimensionless parameters that measure the suitability for SSD-based sorting of a specific batch of pupae are provided. The optimal sorting results are predicted for the highest values of SSD and lowest values of intra-batch variance. Rearing conditions have a strong influence in the performance of the SSD-sorting methods and non-standard rearing can lead to increase pupae size heterogeneity.ConclusionsSex sorting of pupae based on size dimorphism can be achieved with a high performance (% males recovery) and a reasonably high purity (% males on the sorted sample) for the different Aedes species and strains. The purity and performance of a sex sorting operation in the tested Aedes species are linked parameters whose relation can be modeled. The conclusions of this analysis are applicable to all the existing SSD-sorting methods. The efficiency of the SSD-sorting methods can be improved by reducing the heterogeneity of pupae size within rearing containers. The heterogeneity between batches does not strongly affect the quality of the sex sorting, as long as a specific separation threshold is not pre-set before the sorting process. For new developments, we recommend using adaptive and precise threshold selection methods applied individually to each batch or to a mix of batches. Adaptive and precise thresholds will allow the sex-sorting of mixed batches in operational conditions maintaining the target purity at the cost of a reduction in performance. We also recommend a strategy whereby an acceptable level of purity is pre-selected and remains constant across the different batches of pupae while the performance varies from bat...

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