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Understanding host utilization by mosquito vectors is essential to assess the risk of vector-borne diseases. Many studies have investigated the feeding patterns of Culex mosquitoes by molecular analysis of blood-meals from field collected mosquitoes. However, these individual small-scale studies only provide a limited understanding of the complex host-vector interactions when considered in isolation. Here, we analyze the Culex blood-feeding data from 109 publications over the last 15 years to provide a global insight into the feeding patterns of Culex mosquitoes, with particular reference to vectors of currently emerging Culex-borne viruses such as West Nile and Usutu. Data on 29990 blood-meals from 70 different Culex species were extracted from published literature. The percentage of blood-meals on amphibian, avian, human, non-human mammalian, and reptilian hosts was determined for each Culex species. Our analysis showed that feeding patterns were not significantly explained by mosquito species-level phylogeny, indicating that external factors play an important role in determining mosquito feeding patterns. For Cx. quinquefasciatus, 'Cx. pipiens pooled', and Cx. tritaeniorhynchus, feeding patterns were compared across the world's seven biogeographical realms. Culex tritaeniorhynchus, 'Cx. pipiens pooled' and Cx. quinquefasciatus all had significantly varied feeding patterns between realms. These results demonstrate that feeding patterns of Culex mosquitoes vary between species but can also vary between geographically distinct populations of the same species, indicating that regional or population-level adaptations are major drivers of host utilization. Ultimately, these findings support the surveillance of vector-borne diseases by specifying which host groups are most likely to be at risk., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Griep 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.)

The Asian bush mosquito Aedes japonicus is rapidly invading North America and Europe. Due to its potential to transmit multiple pathogenic arthropod-borne (arbo)viruses including Zika virus, West Nile virus, and chikungunya virus, it is important to understand the biology of this vector mosquito in more detail. In addition to arboviruses, mosquitoes can also carry insect-specific viruses that are receiving increasing attention due to their potential effects on host physiology and arbovirus transmission. In this study, we characterized the collection of viruses, referred to as the virome, circulating in Ae. japonicus populations in the Netherlands and France. Applying a small RNA-based metagenomic approach to Ae. japonicus , we uncovered a distinct group of viruses present in samples from both the Netherlands and France. These included one known virus, Ae. japonicus narnavirus 1 (AejapNV1), and three new virus species that we named Ae. japonicus totivirus 1 (AejapTV1), Ae. japonicus anphevirus 1 (AejapAV1) and Ae. japonicus bunyavirus 1 (AejapBV1). We also discovered sequences that were presumably derived from two additional novel viruses: Ae. japonicus bunyavirus 2 (AejapBV2) and Ae. japonicus rhabdovirus 1 (AejapRV1). All six viruses induced strong RNA interference responses, including the production of twenty-one nucleotide-sized small interfering RNAs, a signature of active replication in the host. Notably, AejapBV1 and AejapBV2 belong to different viral families; however, no RNA-dependent RNA polymerase sequence has been found for AejapBV2. Intriguingly, our small RNA-based approach identified an ∼1-kb long ambigrammatic RNA that is associated with AejapNV1 as a secondary segment but showed no similarity to any sequence in public databases. We confirmed the presence of AejapNV1 primary and secondary segments, AejapTV1, AejapAV1, and AejapBV1 by reverse transcriptase polymerase chain reaction (PCR) in wild-caught Ae. japonicus mosquitoes. AejapNV1 and AejapTV1 were found at high prevalence (87-100 per cent) in adult females, adult males, and larvae. Using a small RNA-based, sequence-independent metagenomic strategy, we uncovered a conserved and prevalent virome among Ae. japonicus mosquito populations. The high prevalence of AejapNV1 and AejapTV1 across all tested mosquito life stages suggests that these viruses are intimately associated with Ae. japonicus ., Competing Interests: None declared., (© The Author(s) 2023. Published by Oxford University Press.)