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Background: Ae. aegypti mosquitoes are considered a global threat to public health due to its ability to transmit arboviruses such as yellow fever, dengue, Zika and Chikungunya to humans. The lack of effective arboviral vaccines and etiological treatments make vector control strategies fundamental in interrupting the transmission cycle of these pathogens. This study evaluated Ae. aegypti mosquito populations pre- and post-intervention period with disseminating stations of the larvicide pyriproxyfen to understand its potential influence on the genetic structure and population diversity of these vectors., Methodology/principal Findings: This study was conducted in Manacapuru city, Amazonas, Brazil, where 1,000 pyriproxyfen dissemination stations were deployed and monitored from FEB/2014 to FEB/2015 (pre-intervention) and AUG/2015 to JAN/2016 (post-intervention). Low-coverage whole genome sequencing of 36 individuals was performed, revealing significant stratification between pre- and post-intervention groups (pairwise FST estimate of 0.1126; p-value < 0.033). Tajima's D estimates were -3.25 and -3.07 (both p-value < 0.01) for pre- and post-intervention groups, respectively. Molecular diversity estimates (Theta(S) and Theta(Pi)) also showed divergences between pre- and post-intervention groups. PCA and K-means analysis showed clustering for SNP frequency matrix and SNP genotype matrix, respectively, being both mainly represented by the first principal component. PCA and K-means clustering also showed significant results that corroborate the impact of pyriproxyfen intervention on genetic structure populations of Ae. aegypti mosquitoes., Conclusions/significance: The results revealed a bottleneck effect and reduced mosquito populations during intervention, followed by reintroduction from adjacent and unaffected populations by this vector. We highlighted that low-coverage whole genome sequencing can contribute to genetic and structure population data, and also generate important information to aid in genomic and epidemiological surveillance., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Leles et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Proposing substitutes for Pyriproxyfen (PPF) in the auto-dissemination strategy is essential to ensure the continuity of the strategy in the field, especially in the case of the emergence of populations resistant to this larvicide. One possible substitute among the compounds already in use in Brazil is the larvicide Diflubenzuron (DFB). The equation that defines the proportion of oviposition sites (habitats) contaminated by the auto-dissemination strategy was modified to account for the number of visits required to reach the necessary concentration of DFB for contamination, considering scenarios with varying numbers of oviposition sites and mosquito densities. The dissemination was evaluated in oviposition sites of 2 L, 1.5 L, 1 L, 0.5 L, 0.2 L, and 0.1 L. The minimum concentration of active ingredient (a.i) of DFB required for a commercial product to contaminate at least 50% of oviposition sites was also investigated, along with the impact of other vector control methods, such as the removal/destruction of oviposition sites and the use of insecticides to kill adult 'females, on the auto-dissemination approach. The use of pure DFB compounds enabled contamination efficiency of more than 50% in oviposition sites with a volume of less than 2 L in scenarios with fewer oviposition sites. On the other hand, with the use of the commonly used concentration of the product, similar efficacy was only achieved in oviposition sites of 0.1 L and 0.2 L in medium and high infestation scenarios. Strategies that reduce the number of available oviposition sites work synergistically with the auto-dissemination strategy, making it possible to use less concentrated products and contaminated sites of larger volume. The strategy proved to be resilient in situations of insecticide application according to the concentration of DFB used, abundance of females, and low number of oviposition sites. Increasing the number of dissemination traps on the field also contributes to better results, especially for oviposition sites of 0.5 L and 1 L. The results of the model obtained under the stipulated conditions provide further support for the potential use of DFB as a substitute for PPF in the auto-dissemination strategy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Aedes aegypti and Culex quinquefasciatus are the main urban vectors of arthropod-borne viruses causing human disease, including dengue, Zika, or West Nile. Although key to disease prevention, urban-mosquito control has met only limited success. Alternative vector-control tactics are therefore being developed and tested, often using entomological endpoints to measure impact. Here, we test one promising alternative and assess how three such endpoints perform at measuring its effects., Methods: We conducted a 16-month, two-arm, cluster-randomized controlled trial (CRCT) of mosquito-disseminated pyriproxyfen (MD-PPF) in central-western Brazil. We used three entomological endpoints: adult-mosquito density as directly measured by active aspiration of adult mosquitoes, and egg-trap-based indices of female Aedes presence (proportion of positive egg-traps) and possibly abundance (number of eggs per egg-trap). Using generalized linear mixed models, we estimated MD-PPF effects on these endpoints while accounting for the non-independence of repeated observations and for intervention-unrelated sources of spatial-temporal variation., Results: On average, MD-PPF reduced adult-mosquito density by 66.3% (95% confidence interval, 95% CI: 47.3-78.4%); Cx. quinquefasciatus density fell by 55.5% (95% CI: 21.1-74.8%), and Ae. aegypti density by 60.0% (95% CI: 28.7-77.5%). In contrast, MD-PPF had no measurable effect on either Aedes egg counts or egg-trap positivity, both of which decreased somewhat in the intervention cluster but also in the control cluster. Egg-trap data, therefore, failed to reflect the 60.0% mean reduction of adult Aedes density associated with MD-PPF deployment., Conclusions: Our results suggest that the widely used egg-trap-based monitoring may poorly measure the impact of Aedes control; even if more costly, direct monitoring of the adult mosquito population is likely to provide a much more realistic and informative picture of intervention effects. In our CRCT, MD-PPF reduced adult-mosquito density by 66.3% in a medium-sized, spatially non-isolated, tropical urban neighborhood. Broader-scale trials will be necessary to measure MD-PPF impact on arboviral-disease transmission.