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.

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.

Chagas heart disease (CHD), caused by Trypanosoma cruzi infection, is a significant cause of morbidity and mortality in South and Central America. Enalapril, an angiotensin converting enzyme (ACE) inhibitor, is an important drug used to ameliorate heart functional capacity and its remodelling in individuals presenting CHD. In this study, we evaluated the effects of enalapril on systemic and cardiac immune response during experimental acute CHD. C57BL/6 mice infected with 50 trypomastigote forms of T. cruzi (Colombian strain) were treated daily with enalapril (25 mg/kg) and, after 30 days, a reduction in seric levels of IFN-gamma, TNF-alpha, CCL5/RANTES and nitric oxide, but not in that of IL-10, was detected. This imbalance of cytokines reflects in a reduction of heart mononuclear infiltration and in an increasing of cardiac mast cells. Enalapril also presents a new and interesting in vitro and in vivo anti-T. cruzi activity probably acting on parasite oxidative pathway via cytochrome-P450. Our data show that enalapril exerts an important anti-T. cruzi and anti-inflammatory activity during acute CHD reducing inflammatory cells and, possibly, preventing fibrotic process in the chronic phase. Nevertheless, further studies are still necessary to clarify the mechanisms by which this drug is acting on the parasites and on the immune pathways.

The sterile insect technique (SIT) application, as an alternative tool for conventional mosquito control methods, has recently gained prominence. Nevertheless, some SIT components require further development, such as protocols under large-scale conditions, focusing on packing and shipping mosquitoes, and considering transporting time. Immobilization of Aedes aegypti males was tested at temperatures 4, 7, 10, and 14 °C, and each temperature was assessed for 60, 90, and 120 min. The recovery after 24 h was also studied. Chilled and control-reared males had comparable survival rates for all conditions, although 4 °C for 120 min impacted male survival. The male escape rate was affected after 60 min of exposure at 4 °C; this difference was not significant, with 24 h of recovery. First, we defined the successful immobilization at 4 °C for 60 min, thus enabling the evaluation of two transportation intervals: 6 and 24 h, with the assessment of different compaction densities of 100 and 150 mosquitoes/cm3 at 10 °C to optimize the shipment. Compaction during simulated mosquito shipments reduced survival rates significantly after 6 and 24 h. In the mating propensity and insemination experiments, the sterile males managed to inseminate 40 to 66% for all treatments in laboratory conditions. The male insemination propensity was affected only by the highest compaction condition concerning the control. The analysis of the densities (100 and 150 males/cm3) showed that a higher density combined with an extended shipment period (24 h) negatively impacted the percentage of inseminated females. The results are very helpful in developing and improving the SIT packing and shipment protocols. Further studies are required to evaluate all combined parameters’ synergetic effects that can combine irradiation to assess sexual competitiveness when sterile males are released into the field.

We provide the first evidence of a recent invasion of Aedes aegypti (Linnaeus in Hasselquist, 1762) and Aedes albopictus (Skuse 1894), followed by dengue virus, in tropical montane cities in south-eastern Brazil, Mariana, and Ouro Preto, at mid and high altitudes, respectively. Long-term temperature variation, dengue public data, and sampling of immature and adult mosquitoes (ovitraps and mosquitraps) in contrasting habitats were used to explain the distribution of Aedes in what in these two cities. From 1961 to 2014, the annual temperature increased significantly due to increases in winter temperatures. In the 1990s/2000s, the winter temperature was 1.3°C warmer than in the 1960s, when it varied from 21.2 to 18.9°C. After 2007, the winter temperatures increased and ranged from 21.6 to 21.3°C. The first autochthonous dengue cases in Mariana and Ouro Preto were in 2007, followed by few occurrences until in 2012, when the mean numbers increased three-fold, and peak at 2013. The continuous ‘warmer winter’ may have trigged the Aedes invasion. Aedes species benefited from higher winter temperatures, which was an important driver of their invasion of the state of Minas Gerais in the 1980s and, more recently, in the remaining montane urban habitats in this region. In both 2009 and 2011, we found more Aedes in Mariana than Ouro Preto, and more Ae. albopictus in green areas and Ae. aegypti in houses, the expected pattern for well-established populations.