Monitoring Dengue Virus in Aedes aegypti to Improve Dengue Surveillance and Control in Puerto Rico

<i>Aedes aegypti</i> is the primary urban vector for several important arboviruses, including dengue, chikungunya, yellow fever, and Zika viruses. Traditional dengue virus (DENV) surveillance relies on passive reporting of human cases, which often underestimates transmission due to asymptomatic or unreported infections. This study evaluated the utility of monitoring DENV in <i>Ae. aegypti</i> mosquitoes to improve detection of local dengue transmission and inform vector control strategies during the 2024 dengue epidemic in Puerto Rico. Mosquito surveillance was conducted in 15 neighborhoods within the San Juan metropolitan area where confirmed dengue cases had been recently reported. Adult female <i>Ae. aegypti</i> were collected weekly using Autocidal Gravid Ovitraps (AGO traps) placed within a 200 m radius of index cases. Pools of 1-20 mosquitoes were tested for DENV RNA and serotype using RT-PCR. Surveillance continued for up to 91 days in study areas, depending on virus detection. A total of 29,354 female <i>Ae. aegypti</i> were collected, of which 29,211 females were pooled (1-20 specimens per pool) into 3878 pools and analyzed. DENV was detected in 49 pools across 11 neighborhoods, with serotypes DENV-1, DENV-2, and DENV-3 identified. Multiple serotypes were sometimes detected in mosquitoes from the same neighborhood. Minimum infection rates and vector indices were higher during the epidemic than in previous inter-epidemic periods, and mosquito densities exceeded thresholds considered protective against outbreaks. Entomo-virological surveillance detected a greater variety and evenness of serotypes than passive human surveillance. These findings suggest that entomo-virological surveillance can complement passive case surveillance, providing a more comprehensive detection of DENV circulation. Integrating mosquito-based and human surveillance can improve outbreak detection, guide vector control, and aid in reducing dengue burden in affected communities.