Clinical presentation of proventricular dilatation disease (PDD) cases and controls. A. Necropsy view of control (left panel) African gray parrot (Psittacus erithacus) that died of other causes. The normal-sized proventriculus is not visible in this view as it lies under the left liver lobe (L). Necropsy view of a great green macaw (Ara ambiguus) with PDD (right panel). The proventriculus (PV) is markedly distended and extends laterally well beyond the left lobe of L. The heart (H) is marked for orientation. B. Contrast fluoroscopy view of control (left panel) African gray parrot (Psittacus erithacus) 1.5 hours after administration of barium sulfate. The kidney (K) is marked for orientation. The outline of both the PV and V is clearly visible, with normal size and shape. Within the intestinal loops (IL), wider and thinner sections represent active peristalsis. Right panel, representative PDD case, Eclectus parrot (Eclectus roratus) 18 hours after administration of barium. The PV is markedly distended and contains most of the contrast material, with less in the V and within the IL. A large filling defect (*) representing impacted food material. The kidney (K) is shown for orientation. These findings are typical for PDD; however PDD was not confirmed by histology in this case. C. Proventriculus histopathology. Hematoxylin and eosin staining of proventriculus histological sections from a blue and yellow macaw (Ara ararauna) with PDD. Proventricular gland (G) is shown for orientation. Left panel, normal appearing myenteric ganglion detected within the proventriculus of this case (arrow); right panel, marked lymphoplasmacytic infiltration present within a myenteric ganglion (arrows). Right panel inset, higher magnification. D. CNS histopathology. Hematoxylin and eosin staining of a cerebral section from a control (left panel) African gray parrot (Psittacus erithacus) that died of other causes. Right panel, African gray parrot (Psittacus erithacus) with PDD. Perivascular cuffing is evident around blood vessels (arrows). Inset, higher magnification.

Avian bornavirus (ABV) genome sequence recovery and comparative analysis to Borna disease virus (BDV) genomes. A. Bornaviridae genome schematic. Grey bar at base, non-segmented negative sense viral RNA (vRNA) of Bornaviridae genome; coordinates of major sequence landmarks highlighted below. Green bars and dashed lines, transcription initiation sites (TISs); red bars, transcription termination sites. Distinct ORF-encoding transcription products and the gene products they encode are diagrammed above: TIS1 transcripts encoding nucleocapsid (N) gene, pink; TIS2 transcripts encoding phosphoprotein (P) and X genes, green; TIS3 transcripts encoding the matrix (M), glycoprotein (G) and polymerase (large or 'L') gene, blue. Exons, thick solid black lines; introns, thin solid black lines; dashed black lines, 3'ends of transcripts generated transcription termination read-through; shaded boxes, location of ORFs in transcripts; reading frames for ORFs from multiple genes generated from TIS3 indicated at right. Array probes track, Bornaviridae oligonucleotide 70 mer probes from the Virochip array. PCR primers track, primers generated for PCR follow up and screening of specimens in this study for detection of Bornaviridae species with expected product diagrammed below. vRNA RT-PCR track, overlapping vRNA clones and RACE products recovered directly from RNA extracted from crop tissue of a histologically confirmed case of PDD. Solexa reads track shows distribution of 33 mer reads with at least 15 bp sequence identity to recovered ABV genome sequence. Sequence identity with BDV genomes track shows scanning average pairwise nucleotide sequence identity (window size of 100 nucleotides, advanced in single nucleotide steps) shared between ABV and all BDV genome sequences in NCBI. A dashed line on the graph indicates 50% identity threshold for reference. B. Phylogenetic analysis of ABV genome and the 4 representative BDV genome isolates. Neighbor-joining phylogenetic trees based on nucleotide sequences of the ABV genome sequence [GenBank:EU781967] and the following representative BDV genome sequences: H1766 [GenBank:AJ311523], V/Ref [GenBank:NC_001607], He/80 [GenBank:L27077], and No/98 [GenBank:AJ311524)] Scale bar, genetic distance.

Comparison of sequences recovered from ABV PCR screening to 4 representative genetic isolates of BDV. Neighbor-joining Phylogenetic tree of ABV nucleotide sequences recovered by PCR screening with ABV consensus primers for subsequences within the L gene (A), the M gene (B), or the N gene (C).