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Fig 1.

Taxonomy of the Bornaviridae family.

Complete P gene sequences of representative bornavirus sequences analysed using Neighbor-Joining algorithm and Jukes-Cantor distance model in Geneious R8. Values at branches represent support in 1,000 bootstrap replicates. Only bootstrap values ≥70 at major branches are shown. Bornaviruses marked with asterisks are not classified following the currently accepted taxonomy [2]. ABBV = aquatic bird bornavirus, BoDV = Borna disease virus, CnBV = canary bornavirus, EsBV = estrildid finch bornavirus, GaVV = Gabon viper virus, LGSV = Loveridge´s garter snake virus, MuBV = munia bornavirus, PaBV = parrot bornavirus, VSBV = variegated squirrel bornavirus.

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Table 1.

Sequences of avian bornaviruses available in GenBank.

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Table 2.

Detection of ABBV-1 by RT-PCR from aquatic birds in Germany, 2009–2016.

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Fig 2.

ABBV-1 sequences from wild aquatic birds form continent-associated clusters.

Partial sequences of N (A, 320 bp) or M (B, 200 bp) genes of ABBV-1 were analysed. Phylogenetic analysis was performed using Neighbor-Joining algorithm and Jukes-Cantor distance model in Geneious R8. The trees were rooted with Bornaviridae references sequences (not shown; see Fig 1). Values at branches represent support in 1,000 bootstrap replicates. Only bootstrap values ≥70 at major branches are shown. Nodes with bootstrap support of ≥90 in additionally performed ML analysis are indicated by asterisks. Sequences depicted in bold were generated during this study. Clusters 3 and 4 were confirmed to be independent by alignment of short overlapping partial M gene sequences (113 bp) from both clusters (data not shown).

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Fig 3.

Clusters of PaBV-4 sequences from captive psittacines are not associated with regional origin or host species.

Partial PaBV-4 N gene sequences (326 bp) were analysed using Neighbor-Joining algorithm and Jukes-Cantor distance model in Geneious R8. The tree was rooted with Bornaviridae references sequences (not shown; see Fig 1). Values at branches represent support in 1,000 bootstrap replicates. Only bootstrap values ≥70 at major branches are shown. Nodes with bootstrap support of ≥90 in additionally performed ML analysis are indicated by asterisks. Sequences depicted in bold were generated during this study. Sequences from African grey parrots (Psittacus erithacus) are underlined. (◇) flock A, Germany, 2015; () flock B, Canada, 2009 [43]; (■) flock C, Germany, 2012.

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Fig 4.

PaBV-2 sequences from captive psittacines do not form clear clusters.

Partial PaBV-2 N gene sequences (342 bp) were analysed using Neighbor-Joining algorithm and Jukes-Cantor distance model in Geneious R8. The tree was rooted with Bornaviridae references sequences (not shown; see Fig 1). Values at branches represent support in 1,000 bootstrap replicates. Only bootstrap values ≥70 at major branches are shown. Nodes with bootstrap support of ≥90 in additionally performed ML analysis are indicated by asterisks. Sequences depicted in bold were generated during this study. (*) flock B, Canada, 2009 [43]; (●) flock D1, USA, 2008; (○) flock D2, USA, 2008; flocks D1 and D2 are epidemiologically linked [26].

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Table 3.

Bornavirus detection by RT-PCR from wild and captive Passeriformes in Germany, 2011–2016.

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Fig 5.

Clustering of CnBV-1, CnBV-2 and CnBV-3 detected in common canaries.

Partial N gene sequences (343 bp) were analysed using Neighbor-Joining algorithm and Jukes-Cantor distance model in Geneious R8. The tree was rooted with Bornaviridae references sequences (not shown; see Fig 1). Values at branches represent support in 1,000 bootstrap replicates. Only bootstrap values ≥70 at major branches are shown. Nodes with bootstrap support of ≥90 in additionally performed ML analysis are indicated by asterisks. Sequences depicted in bold were generated during this study. (▲) flock E, Germany, 2014–2015.

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