Wolbachia are endosymbiotic bacteria that infect a wide range of arthropods and filarial nematodes, often manipulating host reproduction. The efficacy of Wolbachia-based interventions for dengue and chikungunya control has been validated through numerous field studies in recent years. This study aimed to investigate the diversity and prevalence of Wolbachia infections in sylvatic mosquitoes from two locations in Recife, Brazil. Multiple mosquito species were screened for Wolbachia using both target marker gene amplification coupled with Sanger sequencing and whole-genome sequencing (WGS) approaches. Phylogenetic analyses were conducted to classify Wolbachia strains into supergroups and assess their evolutionary relationships. Results revealed the presence of Wolbachia in eleven mosquito species examined, with different infection rates. Both supergroups A and B of Wolbachia strains were identified, with Aedes albopictus showing co-infection by both supergroups through the WGS approach. We also detected indirect evidence of Wolbachia horizontal transmission among mosquitoes and other distant host orders. This study provides valuable insights into the distribution and diversity of Wolbachia in sylvatic mosquitoes from Brazil and adds new important data about Wolbachia detection through target marker gene amplicon coupled with Sanger sequencing and WGS methods, highlighting its complementarity to ascertain the presence of Wolbachia in mosquito samples.

Mosquitoes of the Aedes and Culex genera are primary vectors of arboviruses such as the dengue, Zika, chikungunya (CHIKV), Oropouche, and West Nile viruses, causing millions of infections annually. Standard virus detection in mosquitoes requires capturing, transporting, and processing samples with a cold chain to preserve RNA, which is challenging in resource-limited areas. FTA cards preserve viral RNA at room temperature and have been used to collect mosquito saliva, a key sample for assessing transmission. However, most FTA-based traps require electricity or CO₂, limiting use in low-resource settings. This study adapted and evaluated the BR-ArboTrap, a low-cost trap derived from an oviposition trap, integrating a sugar-based attractant with FTA cards to collect mosquito saliva, without electricity or refrigeration. Aedes aegypti exposed to CHIKV were used in three experiments to evaluate: (i) RNA preservation under different conditions, (ii) the minimum number of positive mosquitoes for detection, and (iii) RNA amounts on FTA versus blood. RT-qPCR detected CHIKV RNA in 90% of FTA cards and 96% of exposed mosquitoes. RNA remained stable under varying conditions, with no significant difference compared to blood. BR-ArboTrap is an effective, affordable, and field-ready tool to enhance arbovirus surveillance in remote and low-resource areas.