Table 1.
Western Blot (WB), immunohistochemistry (IHC), and protein-misfolding cyclic amplification results and incubation periods of Tg[CerPrP] mouse bioassay.
Figure 1.
Spongiform degeneration and PrPCWD identified by histopathology and immunohistochemistry.
Vacuolated neurons and spongiform degeneration of the neuropil characteristic of a TSE is evident on H&E staining, with the colocalization of PrPCWD specific immunostaining of florid plaques in the cortices of mice inoculated with positive control inoculum and concentrated urine and saliva from CWD-infected cervids. Negative control mice showed no evidence of spongiform degeneration or PrPCWD immunostaining. HRP-conjugated BAR-224 was used as a primary antibody. (Measure bar, 50 µm).
Figure 2.
Western Blot detection of PrPCWD in urine and saliva-inoculated mice.
Western blotting analysis of control and test mice, demonstrating PrPCWD in positive control mice (lanes 1 and 2), as well as urine (lanes 3 and 4) and saliva (lanes 5 and 6) inoculated mice. Protease-resistant prions were not detected in negative control mice (lanes 7 and 8). Flanking lanes represent undigested PrPC.
Figure 3.
Histopathologic evaluation of renal tissues from donor cervids.
(A) Minimal, chronic and proliferative glomerular disease and (B) mild interstitial fibrosis and lymphocytic infiltration were observed in 4 out of 5 donor deer. The remaining deer showed evidence of mild lymphocytic glomerulonephritis (C) as well as “tubular proteinosis.” (D, arrows).
Figure 4.
Serial PMCA amplification of PrPCWD in concentrated deer urine and in the brains of urine-inoculated mice.
A) PrPCWD was detectable by serial PMCA (sPMCA) in control and urine inocula (lanes 1 and 2, respectively), while PrPCWD could not be identified in saliva and negative control inocula (lanes 3 and 4, respectively) after 3 rounds of amplification. B) Three rounds of sPMCA also amplified PrPCWD in the brains of CWD-infected mice, including positive-control inoculated mice and a single mouse inoculated with lyophilized urine (lanes 1 and 3, respectively). PrPCWD was not amplified in mice inoculated with negative control material (lanes 5 and 6) or in other mice inoculated with either urine (lane 2) or saliva (lane 4) from CWD+ deer. All flanking lanes represent undigested PrPC.