Objective To investigate the species and prevalence of blood-sucking insects and their viruses in Qiongzhong county, Hainan province, China. Methods In August 2019, UV mosquito light-trapas (Gongfu Xiaoshuai) were used to collect specimens in the local environment (sheep pens and pig pens) where blood-sucking insects bred, and dry ice was used to transport the specimens to the laboratory. Virological and molecular biological methods were used to isolate viruses and identify the virus isolates. Results A total of 34 mosquitoes from 3 genera (including 26 from Culex, 1 from Anopheles, and 7 Armigeres subalbatus mosquitoes) and 1 450 midges (to be identified) were collected. A virus isolate (HNQZ1927) was isolated from the collected midge specimens, which could cause cytopathic effect in mosquito cells (C6/36), but did not cause cytopathic effect in BHK-21 cells. The gene amplification and nucleotide sequencing and analysis showed that HNQZ1927 was a 10-gene segment orbivirus. The nucleotides (amino acids) in the coding region of HNQZ1927 had 79.4%-98.7% (94.6%-100%) homology with those of Tibet orbivirus (TIBOV) from the genus Orbivirus, and had 1.2%-78.1% homology with those of other orbiviruses except TIBOV. The phylogenetic analysis of the viral genome showed that the HNQZ1927 virus was in the same evolutionary branch as TIBOV XZ0906, which was previously isolated from An. maculatus collected in Tibet, and the TIBOV isolated from Cx. tritaeniorhynchus, Cx. fatigans, and some species of midges in China. Conclusion The HNQZ1927 virus isolated from midges in Hainan province is TIBOV, which is the first time that TIBOV has been isolated from midge specimens collected from nature in Hainan province, China.
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