New techniques and methods are being sought to try to win the battle against mosquitoes. Recent advances in molecular techniques have led to the development of new and innovative methods of mosquito control based around the Sterile Insect Technique (SIT) [1][2][3] . A control method known as RIDL (Release of Insects carrying a Dominant Lethal) 4 , is based around SIT, but uses genetic methods to remove the need for radiationsterilization [5][6][7][8] . A RIDL strain of Ae. aegypti was successfully tested in the field in Grand Cayman 9,10

Major efforts are currently underway to develop novel, complementary methods to combat mosquito-borne diseases. Mosquito genetic control strategies (GCSs) have become an increasingly important area of research on account of their species-specificity, track record in targeting agricultural insect pests, and their environmentally non-polluting nature. A number of programs targeting Aedes and Anopheles mosquitoes, vectors of human arboviruses and malaria respectively, are currently being developed or deployed in many parts of the world. Operationally implementing these technologies on a large scale however, beyond proof-of-concept pilot programs, is hampered by the absence of adequate sex separation methods. Sex separation eliminates females in the laboratory from male mosquitoes prior to release. Despite the need for sex separation for the control of mosquitoes, there have been limited efforts in recent years in developing systems that are fit-for-purpose. In this special issue of Parasites and Vectors we report on the progress of the global Coordinated Research Program on “Exploring genetic, molecular, mechanical and behavioural methods for sex separation in mosquitoes” that is led by the Insect Pest Control Subprogramme of the Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture with the specific aim of building efficient sex separation systems for mosquito species. In an effort to overcome current barriers we briefly highlight what we believe are the three main reasons why progress has been so slow in developing appropriate sex separation systems: the availability of methods that are not scalable, the difficulty of building the ideal genetic systems and, finally, the lack of research efforts in this area.

In an attempt to control the mosquito-borne diseases yellow fever, dengue, chikungunya, and Zika fevers, a strain of transgenically modified Aedes aegypti mosquitoes containing a dominant lethal gene has been developed by a commercial company, Oxitec Ltd. If lethality is complete, releasing this strain should only reduce population size and not affect the genetics of the target populations. Approximately 450 thousand males of this strain were released each week for 27 months in Jacobina, Bahia, Brazil. We genotyped the release strain and the target Jacobina population before releases began for >21,000 single nucleotide polymorphisms (SNPs). Genetic sampling from the target population six, 12, and 27–30 months after releases commenced provides clear evidence that portions of the transgenic strain genome have been incorporated into the target population. Evidently, rare viable hybrid offspring between the release strain and the Jacobina population are sufficiently robust to be able to reproduce in nature. The release strain was developed using a strain originally from Cuba, then outcrossed to a Mexican population. Thus, Jacobina Ae. aegypti are now a mix of three populations. It is unclear how this may affect disease transmission or affect other efforts to control these dangerous vectors. These results highlight the importance of having in place a genetic monitoring program during such releases to detect un-anticipated outcomes.

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.

The number of mosquito populations resistant to insecticides is increasing along with the reemerging of vector-borne diseases. New technologies are under evaluation to complement the strategies used against these mosquitoes. Transgenic mosquitoes are one approach that some countries are considering and they are being evaluated to control the wild population. Although they have achieved success in population suppression of Aedes aegypti (L.) (Diptera: Culicidae), these studies have not demonstrated what the outcomes are when releases are interrupted (ceased). In this study, after demonstrating suppression of Ae. aegypti using transgenic technology, changes in the spatial distribution of the infestation and the abundance of the vector Ae. aegypti were assessed in the post-release period, along with fluctuation of transgenic mosquitoes in two areas of Brazil. In both pilot trials, there was an average suppression of ca. 70% of the wild population due to the release of transgenic males compared to the pre-release period. In Juazeiro (Mandacaru), in the post-release phase, the number of eggs per trap ranged between 0.06 and 14.41 (mean AE SE = 4.44 AE 0.44), and the ovitrap index (OI = number of ovitraps with eggs/total number of ovitraps recovered) ranged from 0.01 to 0.43 (0.13 AE 0.01). In Jacobina (Pedra Branca), during the post-release phase, the number of eggs per trap ranged between 1 and 7.2 (1.72 AE 0.72), and the OI ranged from 1 to 0.83 (0.095 AE 0.032). The mosquito population in Juazeiro (Mandacaru) remained suppressed for 17 weeks after the release interruption, whereas in Jacobina (Pedra Branca) suppression lasted 32 weeks. In Juazeiro, transgenic larvae were detected up to 5 months after the interruption of the over-flooding releases of transgenic males. In Jacobina, they were found up to 2 months after the release interruption. The number of eggs collected increased 4-5 months after the release interruption, which indicated that the Ae. aegypti population had been re-established after the interruption of releases. The results demonstrate that the technique requires a continuous release in the treated areas, and after suppression, the release rate can be decreased and used as a barrier against external migration.