Abstract
The family Birnaviridae are a group of non-enveloped double-stranded RNA viruses which infect poultry, aquatic animals and insects. This family includes agriculturally important pathogens of poultry and fish. Recently, next-generation sequencing technologies have identified closely related birnaviruses in Culex, Aedes and Anopheles mosquitoes. Using a broad-spectrum system based on detection of long double-stranded RNA, we have discovered and isolated a birnavirus from Aedes notoscriptus mosquitoes collected in northern New South Wales, Australia. Phylogenetic analysis of Aedes birnavirus (ABV) showed that it is related to Rotifer birnavirus, a pathogen of microscopic aquatic animals. In vitro cell infection assays revealed that while ABV can replicate in Aedes-derived cell lines, the virus does not replicate in vertebrate cells and displays only limited replication in Culex- and Anopheles-derived cells. A combination of SDS-PAGE and mass spectrometry analysis suggested that the ABV capsid precursor protein (pVP2) is larger than that of other birnaviruses and is partially resistant to trypsin digestion. Reactivity patterns of ABV-specific polyclonal and monoclonal antibodies indicate that the neutralizing epitopes of ABV are SDS sensitive. Our characterization shows that ABV displays a number of properties making it a unique member of the Birnaviridae and represents the first birnavirus to be isolated from Australian mosquitoes.
Highlights
The family Birnaviridae comprises a group of non-enveloped viruses with bi-segmented, double-stranded RNA genomes
This analysis identified 14 isolates of Liao ning virus (LNV), including one pool co-infected with Alphamesonivirus 1 (Nam Dinh virus, NDiV), which was first reported in Prow et al [28] (Figure 1a)
Next-generation sequencing of the two pools identified a birnavirus-like sequence with 37.52% identity to the RNA-dependent RNA polymerase (RdRP) of Rotifer birnavirus (RBV) by Blastx analysis (e-value 4e-138)
Summary
The family Birnaviridae comprises a group of non-enveloped viruses with bi-segmented, double-stranded RNA (dsRNA) genomes. Birnaviruses are unique among dsRNA viruses as they appear to share structural and phylogenetic characteristics with positive-sense single-stranded RNA ( + ssRNA) viruses [1,2]. Viruses 2020, 12, 1056 virus (BSNV); Aquabirnavirus, encompassing infectious pancreatic necrosis virus (IPNV) and its relatives; and Entomobirnavirus, comprising type species Drosophila X virus (DXV) and related viruses from insects [3]. Rotifer birnavirus (RBV), a pathogen of the rotifer Branchionus plicatilis [4] and Drosophila birnavirus (DBV) which was identified in small RNA sequencing from Drosophila melanogaster, but does not phylogenetically group with the entomobirnaviruses [5]. Segment A encodes a polyprotein which contains the capsid protein precursor pVP2, endopeptidase VP4 and ribonucleoprotein VP3. VP4 is responsible for autoproteolytic processing of the polypeptide to release the three individual proteins
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