Molecular confirmation of an adenovirus in brushtail possums (Trichosurus vulpecula)

Abstract

Partial genome characterisation of a non-cultivable marsupial adenovirus is described. Adenovirus-like particles were found by electron microscopy (EM) in the intestinal contents of brushtail possums (Trichosurus vulpecula) in New Zealand. Using degenerate PCR primers complementary to the most conserved genome regions of adenoviruses, the complete nucleotide sequence of the penton base gene, and partial nucleotide sequences of the DNA polymerase, hexon, and pVII genes were obtained. Phylogenetic analysis of the penton base gene strongly suggested that the brushtail possum adenovirus (candidate PoAdV-1) belongs to the recently proposed genus Atadenovirus. Sequence analysis of the PCR products amplified from the intestinal contents of brushtail possums originating from different geographical regions of New Zealand identified a single genotype. This is the first report of molecular confirmation of an adenovirus in a marsupial.

Fig. 1

Regions of the possum adenovirus genome that were amplified, cloned, and sequenced, compared with the genome of ovine adenovirus isolate 287, the proposed type species of the proposed genus Atadenovirus. (a) PCR products obtained with the degenerate primers (represented by solid lines). (b) The polymerase–penton base clone (represented by a dashed line), from which the 5′-end of the penton base gene was sequenced.

Fig. 2

Distance matrix analysis of penton base amino acid sequences. The final edited alignment had a length of 436 residues. The two established and two proposed genera are designated. Bootstrap values were calculated for 100 data sets and are shown. Virus types are represented by shortened forms of the ICTV abbreviations (Benkö et al., 2000) containing the host designation and type (or strain) number (B, bovine; C, canine; D, duck; F, fowl; Fr, frog; H, human; M, murine; O, ovine; P, porcine; Po, possum; T, turkey). Accessed data and their accession number in GenBank/EMBL database: B3, AF030154; B4, AF036092; C1, U55001; C2, U77082; D1, Y09598; F1, U46933; F9, AF083975; F10, M87008; Fr1, AF224336; H2, J01917; H3, Z29487; H5, M22141; H7, AD001675; H8, AJ249343; H9, AF217407; H12, X73487; H17, AF108105; H19, AF118438, H37, AF118437; H40, L19443; H41, AF105145; M1, U95438; O287, U40837; P3, AJ237815, Po, AF249332 (present paper); T3, AF074946.