CWD bioassay inocula sources.

a<p>Water, feed buckets and bedding from CWD+ deer suites.</p>b<p>Colorado State University deer number 112 (PO brain inoculated deer from previous study<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005916#pone.0005916-Mathiason1" target="_blank">[27]</a>).</p>c<p>Colorado State University deer number 121 (IC brain inoculated deer from previous study <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005916#pone.0005916-Mathiason1" target="_blank">[27]</a>).</p>d<p>Wisconsin Department of Natural Resources, Madison, WI, USA.</p>e<p>Colorado State Veterinary Diagnostic Laboratory, Fort Collins, CO, USA.</p>f<p>Transitioning from pre-clinical to clinical.</p>g<p>Contains brain tissue from deer of pre-clinical and clinical status at terminal field collection.</p

Study design to investigate CWD vertical/maternal transmission.

*<p>Dam inoculated orally (PO) only. All other dams inoculated PO and subcutaneously (SC).</p><p>Late = > ½ time to dam terminal TSE disease.</p><p>Early = < ½ time to dam terminal TSE disease.</p><p>Late gestation = >3.5 months gestation.</p><p>Early gestation = <3.5 months gestation.</p><p>NCD = No clinical disease.</p><p>dpb = days post birth.</p>1<p>Dam’s first conception.</p>2<p>Dam’s second conception.</p>3<p>Dam’s third conception.</p

Summary of naïve deer exposed to inoculum from CWD+ deer—combined with our previous published findings

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005916#pone.0005916-Mathiason1" target="_blank">[27]</a>.</p

Amplifiable CWD prions detected in muntjac dam and fetal tissues.

<p>All panels show representative Western blot results after 5 rounds of sPMCA. Samples considered CWD positive are denoted with an asterisk (*). (A-H) Complete PK digestion of PrP^C is shown in negative deer brain controls (-C) without and with PMCA. PrP^CWD detection is shown in positive deer controls (+C) following PK digestion without and with PMCA. Tissue abbreviations are defined as follows: C = control, MED = mediastinal lymph node, PAR = parotid lymph node, POP = popliteal lymph node, MES = mesenteric lymph node, PLA = placenta, HLN = hepatic lymph node, TE = third eyelid, MAN = mandibular lymph node, TON = tonsil, ILN = ileocecocolic lymph node, RAM = recto-anal mucosa associated lymphoid tissue, SP = spleen, RLN = retropharyngeal lymph node, IL = ileum, TLN = tracheobronchial lymph node, SLN = sublumbar lymph node, OB = obex, MAM = mammary lymph node, CA = caruncle.</p

Western blot results of deer exposed to body secretions and excreta from CWD+ deer.

<p>Western blot demonstration of the typical PK digestion band shift (28–35 kD) associated with prion infection in the brain (medulla oblongata at obex) of deer receiving blood, brain, saliva or environmental exposure, but not urine and feces from CWD-infected donors. Lanes 1–4 represent CWD+/− deer controls (10% brain homogenate of medulla at obex) without and with PK digestion at 25 µg/ml. Lanes 5–12, 10% brain homogenate of blood, brain, urine/feces or saliva inoculated deer without and with PK digestion at 25 µg/ml. Lanes 13–15, brain homogenate from deer environmentally exposed to CWD+ fomites without and with PK digestion at 25 and 50 µg/ml.</p

PrP^CWD detection in CerTgPrP mice inoculated with muntjac CWD.

<p>Western blot detection of PrP^CWD in brains of CerTgPrP mice (n = 9) IC-inoculated with obex from a CWD+ white-tailed deer (lane 2) or CWD+ muntjac (lanes 4–12) following PK digestion. Complete PK digestion of PrP^C is shown in a mouse inoculated with negative deer obex (lane 2).</p

IHC and PMCA CWD detection in offspring born to mock and CWD-infected muntjac.

<p><b>cl</b> = clinical; <b>fair</b> = health condition; <b>*</b> = tonsil biopsy; ^∧ = RAMALT biopsy; <b>term</b> = terminated due to clinical TSE disease; <b>ND</b> = not done; <b>Pla</b> = placenta; <b>RLN</b> = retropharyngeal lymph node; <b>Sp</b> = spleen; <b>Ton</b> = tonsil; <b>Pop</b> = popliteal lymph node; <b>Mam</b> = mammary lymph node; <b>Ob</b> = obex; <b>Mes</b> = mesenteric lymph node; <b>Iln</b> = Ileocecocolic lymph node; <b>RAMALT</b> =  recto-anal mucosa associated lymphoid tissue; <b>I</b> = ileum; <b>Par</b> = parotid lymph node.</p

CWD bioassay inoculation cohorts.

a<p>Water, feed buckets and bedding from CWD+ deer suites.</p

IHC PrP^CWD detection in viable muntjac fawns born to CWD+ dams.

<p>PrP^CWD is demonstrated (red deposits) in tonsilar biopsies collected at 40 days post birth (dpb, A), 465 dpb (C) and 504 dpb (E) and RAMALT biopsies collected at 103 dpb (B), 386 dpb (D) and 504 dpb (F) and confirmed in the terminal tissues of 1 fawn (G, H, I). Absence of PrP^CWD is shown in negative control tissues (J, K, L). Picture objective is 40X (scale bar = 100 µm) with insets at 20X (scale bar = 200 µm).</p

PrP^CWD detection by longitudinal tonsil biopsy and terminal necropsy.

<p>PrP^CWD detection by longitudinal tonsil biopsy and terminal necropsy.</p