Butcherbird polyomavirus isolated from a grey butcherbird (Cracticus torquatus) in Queensland, Australia
Introduction
Members of the family Polyomaviridae are non-enveloped, icosahedral viruses measuring 40–45 nm in diameter (Imperiale and Major, 2007). Their capsids contain a single circular molecule of double stranded DNA of approximately 5000 bp, in which they encode structural proteins (VP1, VP2, VP3, and sometimes others) on one strand and transforming proteins (large, small and occasionally other T-antigens) on the opposite strand (Imperiale and Major, 2007, Johne et al., 2011). Between the start codons of these two genome regions lies a non-coding control region (NCCR), which contains the origin of DNA replication and regulatory sequences (Imperiale and Major, 2007, Johne et al., 2011).
Over the last few years, there has been a significant increase in the number of known polyomavirus (PyV) isolates from a wide range of vertebrate hosts including birds (Johne et al., 2006, Johne and Müller, 2007, Halami et al., 2010), bats (Misra et al., 2009, Fagrouch et al., 2012, Tao et al., 2013), humans (Allander et al., 2007, Gaynor et al., 2007, Feng et al., 2008, van der Meijden et al., 2010, Scuda et al., 2011, Buck et al., 2012, Lim et al., 2012, Siebrasse et al., 2012, Yu et al., 2012, Korup et al., 2013), non-human primates (Johne et al., 2005, Verschoor et al., 2008, Deuzing et al., 2010, Groenewoud et al., 2010, Leendertz et al., 2011), carnivores (Wellehan et al., 2011, Duncan et al., 2013), horses (Renshaw et al., 2012), rodents (Orba et al., 2011), and a dolphin (Anthony et al., 2013). Three genera have now been erected within the family: Orthopolyomavirus and Wukipolyomavirus for the known PyVs of mammals, and Avipolyomavirus for the known PyVs of birds (Johne et al., 2011).
Butcherbirds are mid-sized, black-grey and white, passerine birds classified within the genus Cracticus (Gillies, 1984). The genus includes seven species, all of which are native to Australasia (Kearns et al., 2013). The grey butcherbird, Cracticus torquatus, is an inhabitant of every State and Territory of Australia (Kearns et al., 2013). It is regarded as common, and listed by the IUCN as a species of least concern (http://www.iucnredlist.org).
This manuscript reports the complete genetic characterisation of Butcherbird polyomavirus, a novel member of the genus Avipolyomavirus, in the context of co-infection with an unknown Avipoxvirus species.
Section snippets
Clinical sample
A wild adult grey butcherbird (C. torquatus) of undetermined sex was found at Morayfield, Queensland, Australia (27.13°S; 152.91°E) on 12th July 2009, and taken to the Australia Zoo Wildlife Hospital for treatment of bilateral peri-ocular proliferative cutaneous lesions. Veterinary treatment of this patient (no. 19840) included antibiotic therapy (enrofloxacin 10 mg/kg intramuscularly, every 12 h), analgesia (meloxicam 0.5 mg/kg intramuscularly, every 12 h) and surgical removal under general
Genome amplification and sequencing
The nested PCR for amplifying a wide range of PyVs yielded a single bright band of approximately 230 bp, which Sanger sequencing revealed to be similar, but not identical, to Crow PyV.
Restriction enzyme digestion of the RCA product with BamHI produced no bands, whereas with EcoRI and KpnI, several faint bands were visible (Fig. 1). Although the results were not clear enough to accurately estimate the sizes of the restriction digest fragments, they indicated that a small circular DNA genome was
Discussion
The Butcherbird PyV genome organisation was typical of members of the genus Avipolyomavirus. It had an ORF-X gene with a predicted splice site, which encoded a proline-rich product between the NCCR and late ORFs; an avian PyV large T-antigen binding site in the NCCR, CC(A/T)6GG, rather than the mammalian motif GAGGC; and lacked both the N-terminal nuclear localization signal within VP1, and CxCxxC PP2A binding site within the small T-antigen (Halami et al., 2010). In silico prediction tools
Conflict of interest statement
None to declare.
Acknowledgments
This study was funded by the Murdoch University School of Veterinary and Life Sciences. The authors would like to acknowledge Claude Lacasse for her assistance with collecting the sample for analysis, and Frances Brigg and David Berryman for assistance with sequencing and bioinformatics analysis.
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