Automated misting systems are a convenient way for homeowners or small businesses to control adult mosquitoes. One such system was presented to the Anastasia Mosquito Control District (AMCD) for evaluation to control caged Aedes aegypti. The system consisted of 3 spray tanks, 2 pumps, water level sensors, and flow meters, and was controlled through an Android tablet loaded with dedicated control software. The evaluation of the system included calibration tests, droplet characterization, spray dispersion in the open field, and effectiveness testing using bio-assay cages for mortality assessment. For these tests, a loop of 14 nozzles 4 m apart was connected and held at 1 m height utilizing a total of 120 m tube. All nozzles were arranged in a 16 × 12 m rectangle laid in the East-West direction. Water was sprayed for calibration and droplet size measurements at pressures of 13.0, 15.5, and 18 bar; water and 10 % red dye solution for spray dispersion at 18 bar pressure, and 0.17 % solution of equalizer 20-20 was sprayed at 18 bar pressure for mortality tests. All 3 replicated tests were conducted in the morning between 9:00 and 11:30am. During this time, temperature ranged from 21 to 26 °C, relative humidity from 54 to 95%, and wind speed from 0 - 2 km/hr. The combined flow rate from all 14 nozzles was significantly affected by pressure and was in agreement with the machine-calculated flow rate. There was a similar flow rate from all nozzles, indicated by a standard error of 0.82 mL/min. The droplet characteristics represented by DV0.1, DV0.5, and DV0.9 were not affected by nozzles but decreased with an increase in pressure as expected. The percentage of coverage on the cards, an indicator of spray dispersion, ranged from 20 -100%, and it was found to increase in the direction of the wind. Mosquito mortality showed a similar trend of increasing in the wind direction and ranged from 30 to 100 %. There was no effect of the location of cages on mosquito mortality. These results indicate that the effectiveness of this spray depends upon wind direction. The results, however, may be different when there is no wind, which may be the case during the times these applications are made.

Aedes aegypti and Aedes albopictus are responsible for transmitting major human arboviruses such as Dengue, Zika, and Chikungunya, posing a global threat to public health. The lack of etiological treatments and efficient vaccines makes vector control strategies essential for reducing vector population density and interrupting the pathogen transmission cycle. This study evaluated the impact of long-term pyriproxyfen exposure on the genetic structure and diversity of Ae. aegypti and Ae. albopictus mosquito populations. The study was conducted in Manaus, Amazonas, Brazil, where pyriproxyfen dissemination stations have been monitored since 2014 up to the present day. Double digest restriction-site associated DNA sequencing was performed, revealing that despite significant local population reductions by dissemination stations with pyriproxyfen in various locations in Brazil, focal intervention has no significant impact on the population stratification of these vectors in urban scenarios. The genetic structuring level of Ae. aegypti suggests it is more stratified and directly affected by pyriproxyfen intervention, while for Ae. albopictus exhibits a more homogeneous and less structured population. The results suggest that although slight differences are observed among mosquito subpopulations, intervention focused on neighborhoods in a capital city is not efficient in terms of genetic structuring, indicating that larger-scale pyriproxyfen interventions should be considered for more effective urban mosquito control.

The growing resistance of Aedes aegypti (L.) to conventional insecticides presents a major challenge in arbovirus control, necessitating the exploration of alternative insecticidal chemistries. Spiromesifen, derived from spirocyclic tetronic acids, is widely used against agricultural pests and is crucial in resistance management due to its unique lipid synthesis inhibition. This study evaluates the insecticidal activity of spiromesifen against temephos-resistant Ae. aegypti populations, focusing on larval body weight, volume, biochemical composition, and adult female reproductive potential. Spiromesifen demonstrated effective larvicidal activity, significantly reducing adult emergence. Resistance to spiromesifen was not observed, with resistance ratios (RR50, RR90) ranging from 0.36- to 3.31-fold. Larvae exposed to LC50 showed significant reductions in body weight and volume, and reduced carbohydrate, lipid, and protein contents. Enhanced catalase activity and malondialdehyde levels indicated increased oxidative stress and lipid peroxidation, highlighting its effects on lipid metabolism. Spiromesifen also exhibited sterilizing effects, significantly reducing fecundity and fertility in adult females, thereby impacting Ae. aegypti reproductive capacity. These findings highlight the potential of spiromesifen as a component of integrated vector management strategies, especially in regions with prevalent insecticide resistance in Ae. aegypti, serving as an effective larvicide and impacting adult reproductive outcomes.

Increasing temperature can enhance the geographical spread and behavior of disease vector mosquitoes, exposing vulnerable populations to Aedes-borne viruses and infections. To address this risk, cost-effective and sustained intervention vector control tools are required, such as volatile pyrethroid spatial repellents. This study used a high-throughput screening system toxicity bioassay to determine the discriminating concentrations of transfluthrin-treated filter papers with variable air-drying times exposed to pyrethroid-susceptible Aedes aegypti mosquitoes. At the highest transfluthrin concentration (0.01706%), a significant reduction in mosquito mortality was observed in filter papers air-dried for 24 h compared to those air-dried for 1 h (odds ratio = 0.390, p < 0.001, 95% confidence interval: 0.23-0.66). Conversely, no significant difference in mortality was found between filter papers air-dried for 1 h and those air-dried for 12 h (odds ratio = 0.646, p = 0.107, 95% confidence interval: 0.38-1.10). The discriminating concentration was 2.8-fold higher for transfluthrin-treated filter papers air-dried for 24 h than it was for papers air-dried for 1 h, and it increased 5-fold from 1 h to 336 h of air-drying. These results show that the optimal air-drying period of transfluthrin-treated filter paper is critical, as higher discriminating concentration values may lead to underestimations of insecticide resistance. The instability of transfluthrin-treated papers necessitates the use of the World Health Organization (WHO) bottle bioassay, which is the preferred method for determining mosquito susceptibility to volatile insecticides.

Mosquito Sterile Insect Technique (SIT) programs can be developed in smaller agencies through synchronization of the colony development to take advantage of the natural male early emergence. This paper examined key aspects of Ae. aegypti colony synchronization work, including egg hatching, larval development, and adult emergence to produce sufficient numbers of adult male mosquitoes within a specific timeframe for irradiation and release. Our data indicated that a relatively low percentage of males are required for colony propagation. Additional results highlighted that fresher Ae. aegypti eggs could yield as high as a 93 % hatching success than older eggs when placed under vacuum pressure in yeast infused water for 1.5 h. Eggs that were one-month old hatched (93 %) better than older eggs (0-32 %). A higher egg density in the hatching flask was correlated to a lower hatch rate, and higher larval density was related to unsynchronized pupae and delayed adult emergence. By keeping Ae. aegypti larvae at reasonable density, over 95 % of adults emerged on the first two days of emergence - indicating a high synchronicity. A standardized colony maintenance protocol therefore renders a synchronized larval development and adult male emergence which are critical in SIT programs.

The Dengue virus (DENV), primarily spread by Aedes aegypti and also by Aedes albopictus in some regions, poses significant global health risks. Alternative techniques are urgently needed because the current control mechanisms are insufficient to reduce the transmission of DENV. Introducing Wolbachia pipientis into Ae. aegypti inhibits DENV transmission, however, the underlying mechanisms are still poorly understood. Innate immune effector upregulation, the regulation of autophagy, and intracellular competition between Wolbachia and DENV for lipids are among the theories for the mechanism of inhibition. Furthermore, mainly three immune pathways Toll, IMD, and JAK/STAT are involved in the host for the suppression of the virus. These pathways are activated by Wolbachia and DENV in the host and are responsible for the upregulation and downregulation of many genes in mosquitoes, which ultimately reduces the titer of the DENV in the host. The functioning of these immune pathways depends upon the Wolbachia, host, and virus interaction. Here, we summarize the current understanding of DENV recognition by the Ae. aegypti\'s immune system, aiming to create a comprehensive picture of our knowledge. Additionally, we investigated how Wolbachia regulates the activation of multiple genes associated with immune priming for the reduction of DENV.

Aedes aegypti mosquitoes are major vectors of dengue, chikungunya, and other arboviral diseases. Ae. aegypti\'s capacity to reproduce and to spread disease depends on the female mosquitoes\' ability to obtain blood meals and find water-filled containers in which to lay eggs (oviposit). While humidity sensation (hygrosensation) has been implicated in these behaviors, the specific hygrosensory pathways involved have been unclear. Here, we establish the distinct molecular requirements and anatomical locations of Ae. aegypti Dry Cells and Moist Cells and examine their contributions to behavior. We show that Dry Cell and Moist Cell responses to humidity involve different ionotropic receptor (IR) family sensory receptors, with dry air-activated Dry Cells reliant upon the IR Ir40a, and humid air-activated Moist Cells upon Ir68a. Both classes of hygrosensors innervate multiple antennal sensilla, including sensilla ampullacea near the antennal base as well as two classes of coeloconic sensilla near the tip. Dry Cells and Moist Cells each support behaviors linked to mosquito reproduction but contribute differently: Ir40a-dependent Dry Cells act in parallel with Ir68a-dependent Moist Cells to promote blood feeding, while oviposition site seeking is driven specifically by Ir68a-dependent Moist Cells. Together these findings reveal the importance of distinct hygrosensory pathways in blood feeding and oviposition site seeking and suggest Ir40a-dependent Dry Cells and Ir68a-dependent Moist Cells as potential targets for vector control strategies.

Background: Dengue is a mosquito-borne tropical disease, caused by the Dengue virus (DENV). It has become a severe problem and is a rising threat to public health. In this study, we have evaluated commercial Merilisa i Dengue NS1 Antigen kit (Meril LifeSciences India Pvt. Ltd.) to detect recombinant dengue virus 2 NS1 antigen (rDNS1Ag) and secreted forms of NS1 antigen (sDNS1Ag). Methods: To determine the detection limit of the kit, 100 nanogram (ng) to 0.001 ng rDNS1Ag was tested. The sensitivity and specificity of the kit was determined using recombinant NS1 antigens of all serotypes of DENV and other flaviviruses. For testing sDNS1Ag, the culture supernatant of the Vero cell lines infected with DENV-2 was tested. Further, a spiking experiment was carried out to check the sensitivity of the kit to detect rDNS1Ag in the pools of Aedes aegypti mosquitoes. Results: It was observed that the kit can detect the rDNS1Ag at 1 ng concentration. The kit was sensitive to detect NS1 antigen of DENV-1, DENV-2 and DENV-3 serotypes and specific for detection of only DNS1Ag as it did not cross-react with NS1 antigen of flaviviruses. The kit was sensitive to detect rDNS1Ag in the mosquito pools as well. In addition, the kit was able to detect the sDNS1Ag in Vero cell culture supernatant. Conclusions: Overall, we observed that the Merilisa i Dengue NS1 Ag kit is sensitive and specific for the detection of DNS1Ag both in recombinant and secretory forms.

Herein, we focused on the larvicidal effects and potential mechanisms of 5-ethenyl-2,2\'-bithiophene (5 EB), a compound isolated from Echinops ritro L. on Aedes aegypti larvae. Our results show that 5 EB exhibits pronounced larvicidal activity against A. aegypti larvae, with an LC50 = 0.24 mg/L, considerably lesser than that of the traditional insecticide, rotenone. Observations using fluorescence microscopy, electron microscopy, and imaging flow cytometry demonstrated that 5 EB targets the hemocytes of larvae, leading to the disruption of their intracellular membrane systems. This disruption leads to considerable damage to the cellular structure and function, leading to the death of test subjects. Note that additional investigation into the molecular mechanism of 5 EB\'s action was conducted using transcriptomic analysis. Both GO and KEGG enrichment analyses reported that the differentially expressed genes were predominantly associated with membranes, lysosomes, and catalytic activities. To summarize, this study provides new options for developing new, environmentally friendly, plant-based larvicides for mosquito control.

Cry toxins, produced by the bacterium Bacillus thuringiensis, are of significant agronomic value worldwide due to their potent and highly specific activity against various insect orders. However, some of these pore-forming toxins display specific activity against a range of human cancer cells whilst possessing no known insecticidal activity; Cry41Aa is one such toxin. Cry41Aa has similarities to its insecticidal counterparts in both its 3-domain toxic core structure and pore-forming abilities, but how it has evolved to target human cells is a mystery. This work shows that some insecticidal Cry toxins can enhance the toxicity of Cry41Aa against hepatocellular carcinoma cells, despite possessing no intrinsic toxicity themselves. This interesting crossover is not limited to human cancer cells, as Cry41Aa was found to inhibit some Aedes-active Cry toxins in mosquito larval assays. Here, we present findings that suggest that Cry41Aa shares a receptor with several insecticidal toxins, indicating a stronger evolutionary relationship than their divergent activities might suggest.