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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.

The range of the mosquito Aedes aegypti continues to expand, putting more than two billion people at risk of arboviral infection. The sterile insect technique (SIT) has been used to successfully combat agricultural pests at large scale, but not mosquitoes, mainly because of challenges with consistent production and distribution of high-quality male mosquitoes. We describe automated processes to rear and release millions of competitive, sterile male Wolbachia-infected mosquitoes, and use of these males in a large-scale suppression trial in Fresno County, California. In 2018, we released 14.4 million males across three replicate neighborhoods encompassing 293 hectares. At peak mosquito season, the number of female mosquitoes was 95.5% lower (95% CI, 93.6–96.9) in release areas compared to non-release areas, with the most geographically isolated neighborhood reaching a 99% reduction. This work demonstrates the high efficacy of mosquito SIT in an area ninefold larger than in previous similar trials, supporting the potential of this approach in public health and nuisance-mosquito eradication programs.Mosquitoes are nearly eradicated in three suburbs of California using accurately sorted sterile male mosquitoes.

The radiation-based sterile insect technique (SIT) has successfully suppressed field populations of several insect pest species, but its effect on mosquito vector control has been limited. The related incompatible insect technique (IIT)—which uses sterilization caused by the maternally inherited endosymbiotic bacteria Wolbachia —is a promising alternative, but can be undermined by accidental release of females infected with the same Wolbachia strain as the released males. Here we show that combining incompatible and sterile insect techniques (IIT–SIT) enables near elimination of field populations of the world’s most invasive mosquito species, Aedes albopictus . Millions of factory-reared adult males with an artificial triple- Wolbachia infection were released, with prior pupal irradiation of the released mosquitoes to prevent unintentionally released triply infected females from successfully reproducing in the field. This successful field trial demonstrates the feasibility of area-wide application of combined IIT–SIT for mosquito vector control. A field trial succeeded in eliminating populations of the mosquito Aedes albopictus through inundative mass release of incompatible Wolbachia -infected males, which were also irradiated to sterilize any accidentally-released females, and so prevent population replacement.

Forecasting the impacts of climate change on Aedes-borne viruses-especially dengue, chikungunya, and Zika-is a key component of public health preparedness. We apply an empirically parameterized model of viral transmission by the vectors Aedes aegypti and Ae. albopictus, as a function of temperature, to predict cumulative monthly global transmission risk in current climates, and compare them with projected risk in 2050 and 2080 based on general circulation models (GCMs). Our results show that if mosquito range shifts track optimal temperature ranges for transmission (21.3-34.0°C for Ae. aegypti; 19.9-29.4°C for Ae. albopictus), we can expect poleward shifts in Aedes-borne virus distributions. However, the differing thermal niches of the two vectors produce different patterns of shifts under climate change. More severe climate change scenarios produce larger population exposures to transmission by Ae. aegypti, but not by Ae. albopictus in the most extreme cases. Climate-driven risk of transmission from both mosquitoes will increase substantially, even in the short term, for most of Europe. In contrast, significant reductions in climate suitability are expected for Ae. albopictus, most noticeably in southeast Asia and west Africa. Within the next century, nearly a billion people are threatened with new exposure to virus transmission by both Aedes spp. in the worst-case scenario. As major net losses in year-round transmission risk are predicted for Ae. albopictus, we project a global shift towards more seasonal risk across regions. Many other complicating factors (like mosquito range limits and viral evolution) exist, but overall our results indicate that while climate change will lead to increased net and new exposures to Aedes-borne viruses, the most extreme increases in Ae. albopictus transmission are predicted to occur at intermediate climate change scenarios.

Urbanization processes are increasing globally. Anthropogenic alterations in the environment have profound effects on biodiversity. Decreased biodiversity due to biotic homogenization processes as a consequence of urbanization often result in increased levels of mosquito vector species and vector-borne pathogen transmission. Understanding how anthropogenic alterations in the environment will affect the abundance, richness, and composition of vector mosquito species is crucial for the implementation of effective and targeted mosquito control strategies. We hypothesized that anthropogenic alterations in the environment are responsible for increasing the abundance of mosquito species that are adapted to urban environments such as Aedes aegypti and Culex quinquefasciatus . Therefore, our objective was to survey mosquito relative abundance, richness, and community composition in Miami-Dade County, Florida, in areas with different levels of urbanization. We selected 24 areas, 16 remote areas comprised of natural and rural areas, and 8 urban areas comprised of residential and touristic areas in Miami-Dade County, Florida. Mosquitoes were collected weekly in each area for 24 h for 5 consecutive weeks from August to October 2020 using BG-Sentinel traps baited with dry ice. A total of 36,645 mosquitoes were collected, from which 34,048 were collected in the remote areas and 2,597 in the urban areas. Our results show a clear and well-defined pattern of abundance, richness, and community composition according to anthropogenic modifications in land use and land cover. The more urbanized a given area the fewer species were found and those were primary vectors of arboviruses, Ae. aegypti and Cx. quinquefasciatus .

The development of CRISPR/Cas9 technologies has dramatically increased the accessibility and efficiency of genome editing in many organisms. In general, in vivo germline expression of Cas9 results in substantially higher activity than embryonic injection. However, no transgenic lines expressing Cas9 have been developed for the major mosquito disease vector Aedes aegypti. Here, we describe the generation of multiple stable, transgenic Ae. aegypti strains expressing Cas9 in the germline, resulting in dramatic improvements in both the consistency and efficiency of genome modifications using CRISPR. Using these strains, we disrupted numerous genes important for normal morphological development, and even generated triple mutants from a single injection. We have also managed to increase the rates of homology-directed repair by more than an order of magnitude. Given the exceptional mutagenic efficiency and specificity of the Cas9 strains we engineered, they can be used for high-throughput reverse genetic screens to help functionally annotate the Ae. aegypti genome. Additionally, these strains represent a step toward the development of novel population control technologies targeting Ae. aegypti that rely on Cas9-based gene drives.

Sterile Insect Technique (SIT), Incompatible Insect Technique (IIT) or a combination of the two has become alternative promising vector control approaches. In order to apply these approaches, the targeted mosquitoes need to be sterilized and released. So far, the irradiation of mosquitoes has been conducted at the pupae or adult stages. In this study, we investigated the possibility of applying X-ray irradiation at the egg stage and also assessed the effect on the development and sterility of both wild-type and Wolbachia trans-infected Aedes aegypti mosquito vectors. The eggs of both wild-type and Wolbachia trans-infected lines were irradiated using X-ray at the doses of 1, 3, 5 and 7 Gy. Development of immature stages was observed. For wild-type Ae. aegypti , X-ray irradiation at the doses from 3 Gy decreased the development of the first-instar larvae and increased the development of the third-instar larvae but there was no effect on pupae. For Wolbachia trans-infected ones, a irradiation dose as low as 1 Gy could increase the development of the forth-instar larvae while an irradiation dose of 7 Gy induced significantly high mortality to the pupae ( p < 0.05). To assess sterility, males and females that emerged from irradiated eggs were mated with the non-irradiated ones. Our results showed that an irradiation dose of 7 Gy significantly caused more than 90% sterility in both wild-type males and females ( p < 0.05). However, this irradiation dose could be reduced to 5 Gy to sterilize both males and females infected with Wolbachia . Our findings revealed, for the first time, that applying a low-dose X-ray irradiation at the egg stage could sterilize both wild-type and Wolbachia trans-infected Ae. aegypti when they become adults. Egg irradiation could make the implementation of SIT, IIT or combined SIT/IIT for vector control much more feasible as the sterile eggs are easier to distribute and operate when compared to other developmental stages of mosquitoes.

The yellow fever mosquito, Aedes aegypti, vectors human pathogens. Juvenile hormones (JH) control almost every aspect of an insect’s life, and JH analogs are currently used to control mosquito larvae. Since RNA interference does not work efficiently during the larval stages of this insect, JH regulation of larval development and mode of action of JH analogs are not well studied. To overcome this limitation, we used a multiple single guide RNA-based CRISPR/Cas9 genome-editing method to knockout the methoprene-tolerant (Met) gene coding for a JH receptor. The Met knockout larvae exhibited a black larval phenotype during the L3 (third instar larvae) and L4 (fourth instar larvae) stages and died before pupation. However, Met knockout did not affect embryonic development or the L1 and L2 stages. Microscopy studies revealed the precocious synthesis of a dark pupal cuticle during the L3 and L4 stages. Gene expression analysis showed that Krüppel homolog 1, a key transcription factor in JH action, was down-regulated, but genes coding for proteins involved in melanization, pupal and adult cuticle synthesis, and blood meal digestion in adults were up-regulated in L4 Met mutants. These data suggest that, during the L3 and L4 stages, Met mediates JH suppression of pupal/adult genes involved in the synthesis and melanization of the cuticle and blood meal digestion. These results help to advance our knowledge of JH regulation of larval development and the mode of action of JH analogs in Ae. aegypti.

Over a century has passed since the discovery that the female anopheline mosquito is a vector of malaria, prompting extensive efforts to combat the disease. Vector control remains the primary malaria prevention strategy, accounting for a remarkable 65% reduction in malaria cases between 2000 and 2015. A thorough understanding of the fundamental biology of malaria vectors is essential for national malaria control programmes, governments, and local communities to implement sustainable and effective vector control measures. This article explores the key aspects of mosquito biology, such as adult and juvenile life stages, the intrinsic incubation period, and blood-seeking behaviour, within the context of the mosquito life cycle. By emphasizing the importance of this knowledge, we illustrate how it can inform community-level control initiatives and guide national programmes in designing more effective interventions for vector-borne disease management. Furthermore, the article underscores how researchers can harness these biological insights to develop and deploy innovative tools that have the potential to revolutionize malaria control efforts. Graphical Abstract Illustration of how the mosquito lifecycle could be exploited to control mosquito populations by targeting mosquito developmental stages to support effective malaria control initiatives.

Background In rural south-eastern Tanzania, Anopheles funestus is a major malaria vector, and has been implicated in nearly 90% of all infective bites. Unfortunately, little is known about the natural ecological requirements and survival strategies of this mosquito species. Methods Potential mosquito aquatic habitats were systematically searched along 1000 m transects from the centres of six villages in south-eastern Tanzania. All water bodies were geo-referenced, characterized and examined for presence of Anopheles larvae using standard 350 mLs dippers or 10 L buckets. Larvae were collected for rearing, and the emergent adults identified to confirm habitats containing An. funestus . Results One hundred and eleven habitats were identified and assessed from the first five villages (all < 300 m altitude). Of these, 36 (32.4%) had An. funestus co-occurring with other mosquito species. Another 47 (42.3%) had other Anopheles species and/or culicines, but not An. funestus , and 28 (25.2%) had no mosquitoes. There were three main habitat types occupied by An. funestus, namely: (a) small spring-fed pools with well-defined perimeters (36.1%), (b) medium-sized natural ponds retaining water most of the year (16.7%), and (c) slow-moving waters along river tributaries (47.2%). The habitats generally had clear waters with emergent surface vegetation, depths > 0.5 m and distances < 100 m from human dwellings. They were permanent or semi-permanent, retaining water most of the year. Water temperatures ranged from 25.2 to 28.8 °C, pH from 6.5 to 6.7, turbidity from 26.6 to 54.8 NTU and total dissolved solids from 60.5 to 80.3 mg/L. In the sixth village (altitude > 400 m), very high densities of An. funestus were found along rivers with slow-moving clear waters and emergent vegetation. Conclusion This study has documented the diversity and key characteristics of aquatic habitats of An. funestus across villages in south-eastern Tanzania, and will form an important basis for further studies to improve malaria control. The observations suggest that An. funestus habitats in the area can indeed be described as fixed, few and findable based on their unique characteristics. Future studies should investigate the potential of targeting these habitats with larviciding or larval source management to complement malaria control efforts in areas dominated by this vector species.