Fig 1.
Experimental design describing the administration of CWD-contaminated tissues to coyotes, and sample collection.
A) Schematics of the experimental design described in Nichols et al. [23]. Three coyotes (Canis latrans) were fed a brain extract containing CWD prions from elk, and feces were collected one day before and five days after treatments. Two additional coyotes were treated with a CWD-free elk brain and feces collected as explained before. B) Following 5 days of feces collection, coyotes were sacrificed and multiple tissues were collected and tested for prion content by PMCA.
Fig 2.
PMCA analyses of fecal samples and tissues from coyotes exposed to CWD prions.
A) Western blots depicting PMCA products (third round) of representative fecal samples and coyote tissues exposed (+), or not (-), to CWD prions. All samples were treated with proteinase K, as explained in Material and Methods, with the exception of “PrPC” (brain extracts from tg1536 mice) that were used to control molecular weights and antibody reactivity. Numbers at the right represent molecular weights. The samples depicted in this panel are representative of coyotes exposed (#133, upper panel; #137, middle and lower panel) or not (#134) to CWD prions. B) Summary of the results collected for this study. Each sample was tested in triplicate, by two researchers. “CWD +” and “CWD –“represents animals exposed (+), or not (-), to CWD prions. Positive results in PMCA are color-filled in the table (red: 3/3; orange: 2/3; yellow: 1/3; no fill: 0/3). Fecal samples are denoted by “Day”, and numbers indicate the collection day after treatment (Day 0 represent control samples collected before treatments). N/T: not tested. Coyotes #133, #135 and #137 were fed a brain homogenate containing CWD prions. Coyotes #132 and #134 were fed a CWD-free elk brain homogenate. Raw data for this figure are available in S1 Raw Image. Additional replicates for the samples tested here are shown in S1 Fig.
Fig 3.
Representative data of serial PMCA for fecal samples from a coyote exposed to CWD prions.
Representative analysis of fecal samples collected from Coyote #133 one day before (Day 0) and daily up to 5 days (Day 1-5) after exposing coyotes to CWD prions. All samples were treated with proteinase K, as explained in Material and Methods, with the exception of “PrPC” (brain extracts from tg1536 mice) that were used to control molecular weights and antibody reactivity. PMCA rounds are depicted at the left of each panel (R01-03). Numbers at the right represent molecular weights. The raw data for this figure is available in S2 Raw Image.