Clinical disease following experimental inoculation.

<p>Experimentally inoculated animals were graded by clinical observation for the development of disease consistent with infection with PPRV. Individual animal scores are shown for each sampling point with the standard deviation around the mean being illustrated.</p

PPRV IHC on sections of facial epithelial tissue. a)

<p>Nasal mucosal epithelium (7 dpi). Immunolabelling of lymphoid (arrow) and epithelial cells (open arrow) indicating early PPRV infection of the nasal lamina propria (LP) and mucosal epithelium. Epithelial immunolabelling is noted almost exclusively in basal layers, surrounding LP papillae, with extension into the stratum spinosum; <b>b)</b> Lingual mucosal epithelium (9 dpi) Immunolabelling indicative of early infection in both the basal epithelium and stratum spinosum (arrow) alongside positively labelled immune cells in the LP (open arrow); <b>c)</b> Conjunctival mucosal epithelium (9 dpi). Evidence of advanced epithelial and proprial infection involving a mixed population of inflammatory and epithelial cells around an exocrine gland (arrows). Note in particular the immunolabelling within the proprial lymphoid follicle circumscribed by this gland (open arrow); <b>d)</b> Nasal skin (9 dpi) - marked epithelial infection and erosion (arrow) in and around two hair follicles; <b>e)</b> Labial mucosal epithelium (9 dpi) Following epithelial infection, lymphoid follicles were often seen to have formed in the LP of facial mucosae. Here a large lumber of positively immunolabelled lymphoid cells are seen (arrows); <b>f)</b> Labial mucosal epithelium (9 dpi) with a large epithelial syncytium (arrow) seen in the lower stratum spinosum layer. All scale bars represent 100 µm.</p

Clinical score sheet for assessment of animals infected with PPRV.

<p>When animal reaches a score of 20 they need to be killed on ethical grounds. The decision to euthanase would be based on the following criteria: 1) A score of 4 is achieved in "General Signs“; 2).</p><p>A score of 3 is achieved in "General Signs" for 2 complete, consecutive days and a score of 10 or greater is achieved in other categories; 3) A.</p><p>score of 2 is achieved in "General Signs" for 2 complete, consecutive days and a score of 15 or greater is achieved in other categories. (*Hecker.</p><p>(1983) The sheep as an experimental animal. Academic Press, London; **Smith and Sherman (2009) Goat Medicine, Wiley-Blackwell. Ames,</p><p>Iowa, USA).</p

Molecular detection of viral nucleic acid by RT-PCR.

<p>RNA was extracted from eye swabs or PBLs and subjected to RT-PCR. The PCR product obtained by using the diagnostic primer set F1b-F2d was used as template for the nested PCR (primer set F1–F2). P =  Number of animals positive for the presence of viral nucleic acid; T =  Number of animals tested.</p

Antigen detection within lymphoid tissues at different days post inoculation following challenge with the CI/89 strain of PPRV.

<p>Tissues were taken on days 2, 5, 7, 9 and 21 and antigen detection was assessed by immunohistochemistry (IHC). Results for days 2 and 21 are omitted as all tissues analysed were negative for virus antigen. Average immunolabelling grades are given following analysis of tissues from 3 animals euthanased at each timepoint. Grades are formulated on a result of viral antigen density throughout a uniform tissue type. Sections were graded on three separate occasions, without referring to previous recorded results to help standardise the classification. Immunolabelling grades are defined as: 0 =  No immunolabelling seen; + =  Mild immunolabelling; ++ =  Moderate immunolabelling; +++ =  Marked immunolabelling. Intermediate grades exist between the above four categories to give the analysis a greater degree of flexibility. /− Tissue type not present within section.</p

Conventional, gel-based PCR-assays for the generic detection of all lyssavirus species.

<p>Conventional, gel-based PCR-assays for the generic detection of all lyssavirus species.</p

Real-time PCR-assays for the detection of RABV.

<p>Real-time PCR-assays for the detection of RABV.</p

Conventional, gel-based PCR-assays for the detection rabies virus.

<p>Conventional, gel-based PCR-assays for the detection rabies virus.</p

Conventional, gel-based PCR-assays for the detection of lyssavirus species other than RABV.

<p>Conventional, gel-based PCR-assays for the detection of lyssavirus species other than RABV.</p

Real-time PCR-assays for the detection of lyssavirus species other than RABV.

<p>Real-time PCR-assays for the detection of lyssavirus species other than RABV.</p