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Wild migratory birds are considered the central reservoirs of avian influenza viruses. H12 viruses are one of the 16 hemagglutinin (HA) subtypes of avian influenza viruses and are rarely reported because they are infrequently detected in birds. Consequently, the ecological and genetic profiles of H12 viruses and their adaptation in domestic birds and mammals remain unclear. Here, we found that H12N5 viruses were predominant in the nine identified H12NX subtypes, with the HA (H12) and neuraminidase (NA) (N5) genes showing combination bias in the categorized analysis of subtype combinations (H12 and N1–N9; H1–H12, H14, H15, and N5). These identified H12N5 viruses were primarily detected in birds of Anatidae and Scolopacidae in North America, excluding their possible characterization as chicken or mammalian viruses. The H12N5 viruses were divided into the North American lineage and Eurasian lineage according to their genetic differences, including the HA and NA surface genes and internal genes, although reassortment was observed between the two lineages. We isolated an Eurasian-lineage H12N5 virus from wild ducks in Eastern China, which was one of the 12 identified H12 viruses in China. Infectivity studies indicated that the H12N5 virus is poorly adapted to domestic ducks and chickens, although viral shedding could be detected in both inoculated and contact birds. Additionally, the naturally isolated H12N5 virus did not achieve good replication in mice. These results indicate that the rare subtype of H12 viruses was mainly pooled in wild migratory birds and has an established phylogeography, with low risks of spillover into domestic birds and mammals. [ABSTRACT FROM AUTHOR]

The emergence of novel avian influenza reassortants in wild birds in recent years is a public health concern. However, the viruses that circulate in migratory birds are not fully understood. In this study, we summarized and categorized global H11 avian influenza viruses and reported that waterfowl and shorebirds are the major reservoirs of the identified H11 viruses. The surveillance data of the 35,749 faecal samples collected from wild bird habitats in eastern China over the past seven years revealed a low prevalence of H11 viruses in birds, with a positive rate of 0.067% (24 isolates). The phylogenetic analysis of the twenty viruses indicated that H11 viruses have undergone complex reassortment with viruses circulating in waterfowl and shorebirds. These tested viruses do not acquire mammalian adaptive mutations in their genomes and preferentially bind to avian-type receptors. Experimental infection studies demonstrated that the two tested H11N9 viruses of wild bird origin replicated and transmitted more efficiently in ducks than in chickens, whereas the pigeon H11N2 virus isolated from a live poultry market was more adapted to replicate in chickens than in ducks. In addition, some H11 isolates replicated efficiently in mice and caused body weight loss but were not lethal. Our study revealed the role of waterfowl and shorebirds in the ecology and evolution of H11 viruses and the potential risk of introducing circulating H11 viruses into ducks or chickens, further emphasizing the importance of avian influenza surveillance at the interface of migratory birds and poultry.

Objective: To construct a prokaryotic expression plasmid of DNA G-quadruplex antibody, express it in E.coli BL21 (DE3) bacterial expression system, purify and identify the antibody., Methods: Chemically synthesized BG4 gene of DNA G-quadruplex antibodies was inserted into pSANG10 plasmid to construct DNA G-quadruplex antibody expression vector pSANG10-BG4. BL21 (DE3) as the host strain was utilized for self-induced expression of the protein. Osmotic lysis method was used for collecting this protein. Thereafter, the protein was purified by histidine tag affinity chromatography and identified by SDS-PAGE and Western blotting. The function of this protein was verified in SW480 colon cancer cells., Results: Double enzyme digestion and gene sequencing confirmed that DNA G-quadruplex antibody expression vector was successfully constructed. The relative molecular mass (Mr) of this protein was 30 000 to 37 000. The protein in a soluble form was expressed in the periplasm of BL21. The protein was of the same size as expected., Conclusion: The DNA G-quadruplex antibody has been successfully prepared.