Employing oxford nanopore technologies (ONT) for understanding the ecology and transmission dynamics of flaviviruses in mosquitoes (Diptera: Culicidae) from Eastern Colombia.

Studies focused on identifying the viral species of Flavivirus in vectors are scarce in Latin America and particularly in Colombia. Therefore, the frequency of infection of the Flavivirus genus and its feeding preferences were identified in the mosquito species circulating in the municipality of Puerto Carreño-Vichada, located in the Eastern Plains of Colombia. This was done by sequencing the viral NS5 and vertebrate 12S rRNA genes, respectively, using Oxford Nanopore Technologies (ONT). A total of 1,159 mosquitoes were captured, with the most abundant species being Aedes serratus at 73.6% (n = 853). All the mosquitoes were processed in 230 pools (2-6 individuals) and 51 individuals, where 37.01% (n = 104) were found to be infected with Flavivirus. In these samples, infection by arboviruses of epidemiological importance, such as dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV), was ruled out by PCR. However, through sequencing, infection by different insect-specific viruses (ISFVs) and a medically important virus, West Nile virus (WNV), were identified in a mosquito of the Culex browni species. Additionally, the feeding patterns showed that most species present a generalist behavior. Given the above, conducting entomovirological surveillance studies is crucial, especially in areas of low anthropogenic intervention, due to the high probability that potentially pathogenic viruses could generate spillover events under deforestation scenarios.

Characterizing viral species in mosquitoes (Culicidae) in the Colombian Orinoco: insights from a preliminary metagenomic study.

Mosquitoes (Diptera: Culicidae) are primary vectors of arthropod-borne viruses (arboviruses) that pose significant public health threats. Recent advances in sequencing technology emphasize the importance of understanding the arboviruses and insect-specific viruses (ISVs) hosted by mosquitoes, collectively called the "virome". Colombia, a tropical country with favorable conditions for the development and adaptation of multiple species of Culicidae, offers a favorable scenario for the transmission of epidemiologically important arboviruses. However, entomovirological surveillance studies are scarce in rural areas of the country, where humans, mosquitoes, and animals (both domestic and wild) coexist, leading to a higher risk of transmission of zoonotic diseases to humans. Thus, our study aimed to perform a preliminary metagenomic analysis of the mosquitoes of special relevance to public health belonging to the genera Ochlerotatus, Culex, Limatus, Mansonia, Psorophora, and Sabethes, within a rural savanna ecosystem in the Colombian Orinoco. We employed third-generation sequencing technology (Oxford Nanopore Technologies; ONT) to describe the virome of mosquitoes samples. Our results revealed that the virome was primarily shaped by insect-specific viruses (ISVs), with the Iflaviridae family being the most prevalent across all mosquito samples. Furthermore, we identified a group of ISVs that were common in all mosquito species tested, displaying the highest relative abundance concerning other groups of viruses. Notably, Hanko iflavirus-1 was especially prevalent in Culex eknomios (88.4%) and Ochlerotatus serratus (88.0%). Additionally, other ISVs, such as Guadalupe mosquito virus (GMV), Hubei mosquito virus1 (HMV1), Uxmal virus, Tanay virus, Cordoba virus, and Castlerea virus (all belonging to the Negevirus genus), were found as common viral species among the mosquitoes, although in lower proportions. These initial findings contribute to our understanding of ISVs within mosquito vectors of the Culicidae family in the Eastern Plains of Colombia. We recommend that future research explore deeper into ISV species shared among diverse vector species, and their potential interactions with arboviruses. In addition, we also showed the need for a thorough exploration of the influence of local rural habitat conditions on the shape of the virome in mosquito vectors.