Proactive One-Health surveillance reveals novel orthobunyaviruses and their potential vectors and reservoirs

Mosquito-borne viruses pose a significant, global threat to human and animal health. Traditionally, arthropod-borne virus (arbovirus) surveillance has been retroactive following an outbreak, which limits the ability to predict and prevent outbreaks. We implemented a proactive arboviral surveillance framework in Panama between 2011 and 2024, combining human, virological, entomological, ecological, and genomic surveillance. We screened 599 rodents, 14,586 mosquitoes, and 398 humans with febrile disease, which resulted in the identification and characterization of two novel orthobunyaviruses, provisionally named Aguas Calientes and Aruza viruses. Additionally, we identified active circulation of Madrid virus and a novel reassortant virus (i.e., Matusagaratí) likely derived from small and large genomic segments of the Madrid virus and the medium segment of an unknown virus. We also identified mosquitoes from the Culex genus (i.e., Cx spissipes , Cx. venezuelensis , Cx. pedroi , Cx. vomerifer , and Cx. adamesi ) and Coquillettidia venezuelensis as potential candidate vectors for these viruses. We developed a real-time RT-PCR method to detect these orthobunyaviruses, incorporating it into human, animal, and mosquito surveillance systems. Host-feeding patterns of these mosquitoes suggest that they may act as a bridge between wildlife and livestock, raising concerns about the potential for pathogen spillover between the two. We propose a new framework for Proactive One Health surveillance of potential arbovirus threats and demonstrate its effectiveness in discovering novel arboviruses and identifying their likely transmission vectors and amplifying hosts.