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Tsetse flies are the sole vectors of human African trypanosomiasis throughout sub-Saharan Africa. Both sexes of adult tsetse feed exclusively on blood and contribute to disease transmission. Notable differences between tsetse and other disease vectors include obligate microbial symbioses, viviparous reproduction, and lactation. Here, we describe the sequence and annotation of the 366-megabase Glossina morsitans morsitans genome. Analysis of the genome and the 12,308 predicted protein–encoding genes led to multiple discoveries, including chromosomal integrations of bacterial (Wolbachia) genome sequences, a family of lactation-specific proteins, reduced complement of host pathogen recognition proteins, and reduced olfaction/chemosensory associated genes. These genome data provide a foundation for research into trypanosomiasis prevention and yield important insights with broad implications for multiple aspects of tsetse biology.

BackgroundDiarrhoeal disease is the third leading cause of death in children under 5 years old with domestic flies acting as important mechanical vectors of diarrhoeal pathogens. To assess the effectiveness of a novel house design, “Star home”, and improved toilets in reducing the abundance of domestic flies, potential carriers of diarrhoeal pathogens, a randomized controlled trial was carried out in rural Tanzania.MethodsDomestic fly populations were monitored in 28 randomly selected Star homes and 28 traditional thatched roofs and mud-walled houses over 2 years from January 2022 to December 2023. Flies were sampled in kitchens and toilets using baited-fly traps from 07.00 h to 17.30 h every 7 weeks. To assess the production of flies from toilets, traps were placed over drop holes to collect emerging flies. Duration of external door openings to the kitchens was recorded with data loggers.FindingsOf the 1527 flies collected, 76% were Chrysomya putoria, 16% Musca domestica and 8% Sarcophaga spp. In kitchen collections, there were 46% fewer C. putoria flies [adjusted mean rate ratio (RR) = 0.54] and 69% fewer Sarcophaga spp. (RR = 0.31) in Star homes compared to traditional houses. There was no difference in the abundance of M. domestica in the two study groups. In toilets, there was 49% fewer C. putoria (RR = 0.51), but no difference was observed for other domestic fly species. No flies emerged from Star home toilets compared with a mean of 4.2 flies/trap/day in traditional toilets. During the day, the external doors od Star homes were open for an average of 13.0 min/h less than in traditional houses.ConclusionsStar homes reduced the abundance of domestic flies, apart from houseflies, in the kitchen and there were fewer C. putoria, a putative vector of diarrhoeal diseases, in Star home toilets compared to traditional houses. Changing the design of buildings can contribute to a decline in domestic flies and may lead to a reduction in diarrhoeal diseases.

Viral diseases transmitted via Aedes mosquitoes are on the rise, such as Zika, dengue, and chikungunya. Novel tools to mitigate Aedes mosquitoes-transmitted diseases are urgently needed. We tested whether commercially insecticide-impregnated school uniforms can reduce dengue incidence in school children. We designed a cluster-randomised controlled trial in Thailand. The primary endpoint was laboratory-confirmed dengue infections. Secondary endpoints were school absenteeism; and impregnated uniforms' 1-hour knock-down and 24 hour mosquito mortality as measured by standardised WHOPES bioassay cone tests at baseline and after repeated washing. Furthermore, entomological assessments inside classrooms and in outside areas of schools were conducted. We enrolled 1,811 pupils aged 6-17 from 5 intervention and 5 control schools. Paired serum samples were obtained from 1,655 pupils. In the control schools, 24/641 (3.7%) and in the intervention schools 33/1,014 (3.3%) students had evidence of new dengue infections during one school term (5 months). There was no significant difference in proportions of students having incident dengue infections between the intervention and control schools, with adjustment for clustering by school. WHOPES cone tests showed a 100% knock down and mortality of Aedes aegypti mosquitoes exposed to impregnated clothing at baseline and up to 4 washes, but this efficacy rapidly declined to below 20% after 20 washes, corresponding to a weekly reduction in knock-down and mosquito mortality by 4.7% and 4.4% respectively. Results of the entomological assessments showed that the mean number of Aedes aegypti mosquitoes caught inside the classrooms of the intervention schools was significantly reduced in the month following the introduction of the impregnated uniforms, compared to those collected in classrooms of the control schools (p = 0.04). Entomological assessments showed that the intervention had some impact on the number of Aedes mosquitoes inside treatment schools immediately after impregnation and before insecticidal activity declined. However, there was no serological evidence of protection against dengue infections over the five months school term, best explained by the rapid washing-out of permethrin after 4 washes. If rapid washing-out of permethrin could be overcome by novel technological approaches, insecticide-treated clothes might become a potentially cost-effective and scalable intervention to protect against diseases transmitted by Aedes mosquitoes such as dengue, Zika, and chikungunya. ClinicalTrials.gov NCT01563640.

Background Deltamethrin 62.5 polymer-enhanced suspension concentrate (SC-PE) is one of the World Health Organization-approved insecticides for indoor residual spraying and was recommended to evaluate its residual activity for determination of appropriate spray cycles in different eco-epidemiologic settings. In the current study, efficacy of deltamethrin 62.5 SC-PE was evaluated against vectors of malaria and its impact on malaria incidence in a Plasmodium falciparum hyper-endemic area in Koraput district, Odisha State, India. Methods The trial had two comparable arms, arm 1 with residual spraying of deltamethrin 62.5 SC-PE and arm 2 with deltamethrin 2.5% WP (positive control). Comparative assessment of the impact of each intervention arm on entomological (density, parity, infection and human blood index), epidemiological (malaria incidence) parameters, residual efficacy and adverse effects were evaluated. Results Both the arms were comparable in terms of entomological and epidemiological parameters. While, deltamethrin 62.5 SC-PE was found to be effective for 150 days in mud and wood surfaces and 157 days in cement surfaces; deltamethrin 2.5% was effective only for 105 days on mud surfaces and 113 days on cement and wood surfaces. Conclusions Deltamethrin 62.5 SC-PE had prolonged killing effectiveness up to 5 months. Hence, one round of IRS with deltamethrin 62.5 SC-PE would be sufficient to cover two existing malaria peak transmission seasons (July–August and October–November) in many parts of India.

Insects enter every passable space on the planet. Despite our best efforts, flying insects infiltrate slightly open windows in domiciles, automobiles, storage spaces, and more. Is this ubiquitous experience a consequence of insect abundance and probability, or are flying insects adept at detecting passageways? There remains a lack of understanding of insect effectiveness in finding passage through the voids and imperfections in physical barriers in response to attractants, a topic particularly critical to the area of insect-borne disease control. In this study, we recorded the passage of Aedes aegypti mosquitoes through voids in vertically oriented bed net fabrics within a cylindrical flight arena. We model the probability mosquitoes will discover and navigate the void in response to a physical attractant by observing their search behavior and quantifying the region within a void that is physically navigable, constrained by body size. Void passage rates were lower than that would be expected by purely randomized search behaviors and decline rapidly as the void diameter approaches the in-flight width of the insect.

Background Mosquito lifespan can influence the circulation of disease causing pathogens because it affects the time available for infection and transmission. The life-cycle of mosquitoes is determined by intrinsic and environmental factors, which can include the availability of hosts and suitable resting environments that shelter mosquitoes from extreme temperature and desiccating conditions. This study determined the parity rates (an indirect measure of survival) and plant resting preference of vectors of Rift Valley fever (RVF) in northeastern Kenya. Methods Resting mosquitoes were trapped during the rainy and the dry season using a Prokopack aspirator from vegetation, whereas general adult populations were trapped using CDC light traps. At each site, sampling was conducted within a 1 km2 area, subdivided into 500 × 500 m quadrants and four 250 × 250 m sub-quadrants from which two were randomly selected as sampling units. In each sampling unit, plants were randomly selected for aspiration of mosquitoes. Only Aedes mcintoshi and Ae. ochraceus were dissected to determine parity rates while all mosquito species were used to assess plant resting preference. Results Overall, 1124 (79 %, 95 % CI = 76.8–81.1 %) mosquitoes were parous. There was no significant difference in the number of parous Ae. mcintoshi and Ae. ochraceus. Parity was higher in the rainy season than in the dry season. Daily survival rate was estimated to be 0.93 and 0.92 among Ae. ochraceus and Ae. mcintoshi, respectively. Duosperma kilimandscharicum was the most preferred plant species with the highest average capture of primary (3.64) and secondary (5.83) vectors per plant, while Gisekia africana was least preferred. Conclusion Survival rate of each of the two primary vectors of RVF reported in this study may provide an indication that these mosquitoes can potentially play important roles in the circulation of diseases in northern Kenya. Resting preference of the mosquitoes in vegetation may influence their physiology and enhance longevity. Thus, areas with such vegetation may be associated with an increased risk of transmission of arboviruses to livestock and humans.

Wolbachia used to counter dengue fever The mosquito-borne viral disease dengue fever is an increasing problem in tropical and subtropical regions. Traditional control measures aimed at reducing populations of the main transmission vector, Aedes aegypti , have had little success. Two papers in this issue report an alternative approach to mosquito population control using the bacterium Wolbachia pipientis , natural insect symbionts that facilitate their own transmission through a process called cytoplasmic incompatibility. In the first paper, Scott O'Neill and colleagues describe a Wolbachia strain derived from fruitflies that significantly reduces dengue virus carriage in mosquitoes without imposing a fitness cost. In the second paper, they demonstrate in a controlled field trial that the release of colonized mosquitoes leads to successful invasion of natural mosquito populations. These results suggest a viable strategy to control dengue fever. Dengue fever is the most important mosquito-borne viral disease of humans with more than 50 million cases estimated annually in more than 100 countries 1 , 2 . Disturbingly, the geographic range of dengue is currently expanding and the severity of outbreaks is increasing 2 , 3 , 4 . Control options for dengue are very limited and currently focus on reducing population abundance of the major mosquito vector, Aedes aegypti 5 , 6 . These strategies are failing to reduce dengue incidence in tropical communities and there is an urgent need for effective alternatives. It has been proposed that endosymbiotic bacterial Wolbachia infections of insects might be used in novel strategies for dengue control 7 , 8 , 9 . For example, the w MelPop-CLA Wolbachia strain reduces the lifespan of adult A. aegypti mosquitoes in stably transinfected lines 8 . This life-shortening phenotype was predicted to reduce the potential for dengue transmission. The recent discovery that several Wolbachia infections, including w MelPop-CLA, can also directly influence the susceptibility of insects to infection with a range of insect and human pathogens 9 , 10 , 11 has markedly changed the potential for Wolbachia infections to control human diseases. Here we describe the successful transinfection of A. aegypti with the avirulent w Mel strain of Wolbachia , which induces the reproductive phenotype cytoplasmic incompatibility with minimal apparent fitness costs and high maternal transmission, providing optimal phenotypic effects for invasion. Under semi-field conditions, the w Mel strain increased from an initial starting frequency of 0.65 to near fixation within a few generations, invading A. aegypti populations at an accelerated rate relative to trials with the w MelPop-CLA strain. We also show that w Mel and w MelPop-CLA strains block transmission of dengue serotype 2 (DENV-2) in A. aegypti , forming the basis of a practical approach to dengue suppression 12 .

Wolbachia used to counter dengue fever The mosquito-borne viral disease dengue fever is an increasing problem in tropical and subtropical regions. Traditional control measures aimed at reducing populations of the main transmission vector, Aedes aegypti , have had little success. Two papers in this issue report an alternative approach to mosquito population control using the bacterium Wolbachia pipientis , natural insect symbionts that facilitate their own transmission through a process called cytoplasmic incompatibility. In the first paper, Scott O'Neill and colleagues describe a Wolbachia strain derived from fruitflies that significantly reduces dengue virus carriage in mosquitoes without imposing a fitness cost. In the second paper, they demonstrate in a controlled field trial that the release of colonized mosquitoes leads to successful invasion of natural mosquito populations. These results suggest a viable strategy to control dengue fever. Genetic manipulations of insect populations for pest control have been advocated for some time, but there are few cases where manipulated individuals have been released in the field and no cases where they have successfully invaded target populations 1 . Population transformation using the intracellular bacterium Wolbachia is particularly attractive because this maternally-inherited agent provides a powerful mechanism to invade natural populations through cytoplasmic incompatibility 2 . When Wolbachia are introduced into mosquitoes, they interfere with pathogen transmission and influence key life history traits such as lifespan 3 , 4 , 5 , 6 . Here we describe how the w Mel Wolbachia infection, introduced into the dengue vector Aedes aegypti from Drosophila melanogaster 7 , successfully invaded two natural A. aegypti populations in Australia, reaching near-fixation in a few months following releases of w Mel-infected A. aegypti adults. Models with plausible parameter values indicate that Wolbachia -infected mosquitoes suffered relatively small fitness costs, leading to an unstable equilibrium frequency <30% that must be exceeded for invasion. These findings demonstrate that Wolbachia -based strategies can be deployed as a practical approach to dengue suppression with potential for area-wide implementation.

The sterile insect technique (SIT) is an environmentally safe and proven technology to suppress wild populations. To further advance its utility, a novel CRISPR-based technology termed precision guided SIT (pgSIT) is described. PgSIT mechanistically relies on a dominant genetic technology that enables simultaneous sexing and sterilization, facilitating the release of eggs into the environment ensuring only sterile adult males emerge. Importantly, for field applications, the release of eggs will eliminate burdens of manually sexing and sterilizing males, thereby reducing overall effort and increasing scalability. Here, to demonstrate efficacy, we systematically engineer multiple pgSIT systems in Drosophila which consistently give rise to 100% sterile males. Importantly, we demonstrate that pgSIT-generated sterile males are fit and competitive. Using mathematical models, we predict pgSIT will induce substantially greater population suppression than can be achieved by currently-available self-limiting suppression technologies. Taken together, pgSIT offers to potentially transform our ability to control insect agricultural pests and disease vectors. Sterile Insect Technique (SIT) is used to suppress wild populations. Here the authors integrate CRISPR-based technology and SIT to develop a precision guided SIT (pgSIT), and demonstrate its proof-of-principle by generating 100% sterile males.

Cucumber mosaic virus (CMV) often accompanies a short RNA molecule called a satellite RNA (satRNA). When infected with CMV in the presence of Y-satellite RNA (Y-sat), tobacco leaves develop a green mosaic, then turn yellow. Y-sat has been identified in the fields in Japan. Here, we show that the yellow leaf colour preferentially attracts aphids, and that the aphids fed on yellow plants, which harbour Y-sat-derived small RNAs (sRNAs), turn red and subsequently develop wings. In addition, we found that leaf yellowing did not necessarily reduce photosynthesis, and that viral transmission was not greatly affected despite the low viral titer in the Y-sat-infected plants. Y-sat-infected plants can therefore support a sufficient number of aphids to allow for efficient virus transmission. Our results demonstrate that Y-sat directly alters aphid physiology via Y-sat sRNAs to promote wing formation, an unprecedented survival strategy that enables outward spread via the winged insect vector. The cucumber mosaic virus is accompanied by short RNA molecules, satellite RNAs. This study shows that leaves infected with Y-satellite RNA preferentially attract aphids and manipulate aphid physiology to promote their spread to neighbouring plants.