Objective:To elucidate the complete genomic characteristics of the Akabane virus (AKV) DHL10M117 strain and the Tibet orbivirus (TIBOV) DH10M1019 strain, isolated from mosquito specimens collected in 2010 from Dehong prefecture, Yunnan province.Methods:The complete RNA virus sequences were obtained using metatranscriptomics and high-throughput sequencing.Results:The complete genome sequences of the DHL10M117 strain, consisting of the S, M, and L gene segments with lengths of 856 bp, 4 309 bp, and 6 869 bp, respectively, were acquired. Phylogenetic analysis indicated that the DHL10M117 strain is an AKV strain, closely related to the AKV strain DHL10M110, isolated from Yunnan. The S and M segment phylogenetic trees revealed that the strain is closely related to AKV strains circulating in southern China and Japan, and distantly related to strains from Australia, suggesting that this strain belongs to the Asian lineage with distinct regional characteristics. Homology analysis confirmed that the gene sequences of the S, M, and L segments of the DHL10M117 strain showed 100% nucleotide and amino acid identity with the DHL10M110 strain. Additionally, the complete genome sequences of the DH10M1019 strain, comprising ten gene segments (Seg-1 to Seg-10) with lengths of 3 950 bp, 2 904 bp, 2 769 bp, 1 978 bp, 1 772 bp, 1 638 bp, 1 165 bp, 1 142 bp, 1 103 bp, and 832 bp respectively, were obtained. Phylogenetic analysis showed that DH10M1019 is a TIBOV strain, with eight gene segments (Seg-1 to Seg-4, Seg-6, Seg-8 to Seg-10) clustering with known strains in the same major branch, while Seg-5 and Seg-7 formed distinct branches, independent of known reference strains, suggesting that DH10M1019 may represent a new serotype of TIBOV.Conclusions:Through complete genome sequence analysis, it was confirmed that DHL10M117 and DH10M1019 are AKV and TIBOV, which provided a scientific basis for the epidemiological characterisation, pathogenicity analysis and investigation of the two viruses.

Objective:The complete genome sequence characteristics of Quang Binh virus (QBV) and Manglie virus (MaV) isolated from mosquitoes in Daluo Town, Menghai county.Methods:Mosquitoes collected in Daluo Town, Menghai county in July 2012 were used for virus isolation. The nucleic acid was extracted from the supernatant of Aedes albopictus cell line (C6/36) showing cytopathic effect (CPE) positivity, and was sent to a company for sequencing after library construction. Phylogenetic and nucleotide/amino acid sequence similarity analysis was performed using DNAStar, Maff, and other softwares.Results:RNA libraries of strains BNDL1205 and BNDL1227 yielded 67 336 692 and 61 259 266 qualified gene sequences (reads) respectively. After assembly and alignment, sequences of lengths 10 865 bp and 10 864 bp were obtained. Sequence analysis indicated that they belong to QBV, with strains BNDL1205 and BNDL1227 clustering with QBV (strain VN180) isolated from Vietnam on the same evolutionary branch, sharing nucleotide similarity of 84.2% and 84.1%, and amino acid similarity of 94.6% and 94.4% respectively. RNA library of strain BNDL1223 yielded 48 622 610 qualified reads. After assembly and alignment, three gene fragments (Contigs) matched MaV. Further merging using SeqMan produced a complete nucleotide sequence of 9 219 bp. Analysis revealed that strain BNDL1223 is closely related to MaV isolated in Yunnan in 2018, sharing nucleotide similarity of 97.8% and amino acid similarity of 99.2%.Conclusions:During the investigation of arboviruses in mosquitoes in Daluo Town, Menghai county, three strains of viruses were identified: 2 strains of QBV and 1 strain of MaV. Local mosquitoes play a significant role in the transmission of QBV and MaV, necessitating enhanced monitoring and detection of local vector mosquitoes.

Objective:To elucidate the complete genomic characteristics of the Akabane virus (AKV) DHL10M117 strain and the Tibet orbivirus (TIBOV) DH10M1019 strain, isolated from mosquito specimens collected in 2010 from Dehong prefecture, Yunnan province.Methods:The complete RNA virus sequences were obtained using metatranscriptomics and high-throughput sequencing.Results:The complete genome sequences of the DHL10M117 strain, consisting of the S, M, and L gene segments with lengths of 856 bp, 4 309 bp, and 6 869 bp, respectively, were acquired. Phylogenetic analysis indicated that the DHL10M117 strain is an AKV strain, closely related to the AKV strain DHL10M110, isolated from Yunnan. The S and M segment phylogenetic trees revealed that the strain is closely related to AKV strains circulating in southern China and Japan, and distantly related to strains from Australia, suggesting that this strain belongs to the Asian lineage with distinct regional characteristics. Homology analysis confirmed that the gene sequences of the S, M, and L segments of the DHL10M117 strain showed 100% nucleotide and amino acid identity with the DHL10M110 strain. Additionally, the complete genome sequences of the DH10M1019 strain, comprising ten gene segments (Seg-1 to Seg-10) with lengths of 3 950 bp, 2 904 bp, 2 769 bp, 1 978 bp, 1 772 bp, 1 638 bp, 1 165 bp, 1 142 bp, 1 103 bp, and 832 bp respectively, were obtained. Phylogenetic analysis showed that DH10M1019 is a TIBOV strain, with eight gene segments (Seg-1 to Seg-4, Seg-6, Seg-8 to Seg-10) clustering with known strains in the same major branch, while Seg-5 and Seg-7 formed distinct branches, independent of known reference strains, suggesting that DH10M1019 may represent a new serotype of TIBOV.Conclusions:Through complete genome sequence analysis, it was confirmed that DHL10M117 and DH10M1019 are AKV and TIBOV, which provided a scientific basis for the epidemiological characterisation, pathogenicity analysis and investigation of the two viruses.

Objective:The complete genome sequence characteristics of Quang Binh virus (QBV) and Manglie virus (MaV) isolated from mosquitoes in Daluo Town, Menghai county.Methods:Mosquitoes collected in Daluo Town, Menghai county in July 2012 were used for virus isolation. The nucleic acid was extracted from the supernatant of Aedes albopictus cell line (C6/36) showing cytopathic effect (CPE) positivity, and was sent to a company for sequencing after library construction. Phylogenetic and nucleotide/amino acid sequence similarity analysis was performed using DNAStar, Maff, and other softwares.Results:RNA libraries of strains BNDL1205 and BNDL1227 yielded 67 336 692 and 61 259 266 qualified gene sequences (reads) respectively. After assembly and alignment, sequences of lengths 10 865 bp and 10 864 bp were obtained. Sequence analysis indicated that they belong to QBV, with strains BNDL1205 and BNDL1227 clustering with QBV (strain VN180) isolated from Vietnam on the same evolutionary branch, sharing nucleotide similarity of 84.2% and 84.1%, and amino acid similarity of 94.6% and 94.4% respectively. RNA library of strain BNDL1223 yielded 48 622 610 qualified reads. After assembly and alignment, three gene fragments (Contigs) matched MaV. Further merging using SeqMan produced a complete nucleotide sequence of 9 219 bp. Analysis revealed that strain BNDL1223 is closely related to MaV isolated in Yunnan in 2018, sharing nucleotide similarity of 97.8% and amino acid similarity of 99.2%.Conclusions:During the investigation of arboviruses in mosquitoes in Daluo Town, Menghai county, three strains of viruses were identified: 2 strains of QBV and 1 strain of MaV. Local mosquitoes play a significant role in the transmission of QBV and MaV, necessitating enhanced monitoring and detection of local vector mosquitoes.

Objective:To investigate the infection status of Kaeng Khoi virus (KKV) in bat flies in Yunnan province.Methods:Specimens of the ectoparasitic bat flies on bats in Baoshan city in 2014 and Ruili city in 2017 were collected and identified, virus isolation was performed by cell culture. The positive isolates were amplified and sequenced to obtain the complete genome sequences, and homology comparison and phylogenetic analysis were carried out.Results:Three positive isolates (WDBC1704, WDBC1710 and BSBC1406) were identified from bat flies in Ruili city and Baoshan city. The supernatant was inoculated into BHK-21 cells for virus isolation, all of which could cause cytopathic effects (CPE) included shrinking, rounding and falling off of BHK-21 cells. The complete genome sequences of the three positive isolates were produced by RT-PCR and high-throughput sequencing. Phylogenetic analysis showed that the S, M, and L genes of all three positive isolates were in the same evolutionary branch as the original KKV strain PSC-19 isolated in Thailand. In this study, three KKV strains isolated were most closely related to the previous isolate WDBC1403 from Yunnan province, of which the BSBC1406 strain formed a small independent branch. The sequence comparison showed that there were significant differences in the M gene among the three isolates. The similarity between BSBC1406 and the other two strains (WDBC1704 and WDBC1710) was only 78.0%-78.1%, the amino acid similarity of the Gc protein in the open reading frame region was 85.3%.Conclusions:Three KKV strains from bat flies were quite different from the prototype strain, especially the BSBC1406 strain was somewhat different from other isolates and may have undergone variation. Surveillance of KKV carried by vector insects in Yunnan province and its relationship with human diseases should be strengthened.

Objective:To study the whole genome and genetic evolution characteristics of the Banna virus carried by Culicoides in Cangyuan County. Methods:Culicoides were collected in Cangyuan county in 2018. After grinding, the supernatant was inoculated onto BHK-21 cells for virus isolation. At the same time, the virus carrying status of Culicoides was detected by metatranscriptomics technology. Perl, Mafft and other bioinformatics softwares were used for sequence alignment, homology and molecular genetic evolution analysis. Results:Through metatranscriptomics method, the whole genome of Banna virus CYC1801 strain was obtained. Analysis of the whole genome showed that segment 4 and segment 7 are closely related to the Vietnamese strains, other segments are closely related to China strains. According to the phylogenetic analysis of the segment 12 gene, CYC1801 and the A2 gene subtype are in the same evolutionary branch and belong to the A2 gene subtype of Banna virus.Conclusions:Banna virus was carried by Culicoides collected in Cangyuan county, which may cause disease epidemic as a vector, so monitoring and prevention should be strengthened.