Infectious prion diseases 1 – scrapie of sheep 2 and chronic wasting disease (CWD) of several species in the deer family 3,4 – are transmitted naturally within affected host populations. Although several possible sources of contagion have been identified in excretions and secretions from symptomatic animals 5–8, the biological importance of these sources in sustaining epidemics remains unclear. Here we show that asymptomatic CWD-infected mule deer (Odocoileus hemionus) excrete CWD prions in their feces long before they develop clinical signs of prion disease. Intracerebral (i.c.) inoculation of irradiated deer feces into transgenic (Tg) mice overexpressing cervid PrP revealed infectivity in 14 of 15 fecal samples collected from 5 deer at 7–11 months before the onset of neurological disease. Although prion concentrations in deer feces were considerably lower than in brain tissue from the same deer collected at the disease terminus, the estimated total infectious dose excreted in feces by an infected deer over the disease course may approximate the total contained in brain tissue. Prolonged fecal prion excretion by infected deer provides a plausible natural mechanism that might explain the high incidence and efficient horizontal transmission of CWD within deer herds 3,4,9, as well as prion transmission between susceptible deer species.

The utility of rectal lymphoid tissue sampling for the diagnosis of chronic wasting disease (CWD) infections in mule deer (Odocoileus hemionus) and white-tailed deer (Odocoileus virginianus) was evaluated. CWD-associated prion protein (PrPCWD) deposits were observed in the rectal mucosa from 19 orally inoculated mule deer by 381 days post-inoculation (p.i.); similarly, 45 out of 50 naturally infected mule deer had PrPCWD in their rectal mucosa. In orally inoculated white-tailed deer, the presence of glycine (G) or serine (S) at codon 96 of the native PrP (denoted 96GG, 96GS or 96SS) appeared to influence the temporal patterns of PrPCWD deposition: nine out of 11 infected 96GG individuals had PrPCWD in their rectal mucosa by 342 days p.i., whereas only three out of seven infected 96GS individuals had PrPCWD in their rectal mucosa by 381 days p.i. and none of three 96SS individuals had PrPCWD in their rectal mucosa by 751 days p.i. These findings support further evaluation of rectal mucosa sampling in CWD surveillance.

Chemical immobilization of wildlife often includes opioids or cyclohexamines. These substances are problematic as a result of their required storage, handling, and record-keeping protocols. A potentially useful alternative sedation protocol includes a combination of butorphanol, azaperone, and medetomidine (BAM: 0.43 mg/kg butorphanol, 0.36 mg/kg azaperone, 0.14 mg/kg medetomidine). One risk of wildlife immobilization with any drug combination is hypoxemia. This may be of particular importance when using an alpha 2 agonist such as medetomidine because of its powerful vasoconstrictive effect. In this prospective study, the BAM combination was evaluated for chemical immobilization of white-tailed deer. Additionally, selected physiologic parameters associated with BAM immobilization, including oxygen saturation via pulse oximetry and arterial blood gas measurement, with and without nasal insufflation of oxygen at a relatively low flow of 3 L/min, were evaluated. The BAM combination resulted in a predictable onset of sedation, with a mean induction time to lateral recumbency of 9.8 +/- 3.6 min. All deer recovered smoothly within a range of 5-20 min after reversal with intramuscular administration of naltrexone, atipamazole, and tolazoline (NAT). Clinically relevant decreases in arterial partial pressure of oxygen (PaO2) and oxygen saturation (SpO2) were observed in animals not receiving supplemental oxygen, while both parameters significantly improved for oxygen-supplemented deer. Pulse oximetry with this protocol was an unreliable indicator of oxygen saturation. In this study, altitude, recumbency, hypoventilation, butorphanol- and medetomidine-specific effects, as well as the potential for alpha 2 agonist-induced pulmonary changes all may have contributed to the development of hypoxemia. Overall, capture of white-tailed deer with the BAM/NAT protocol resulted in excellent chemical immobilization and reversal. Because the BAM combination caused significant hypoxemia that is unreliably detected by pulse oximetry but that may be resolved with nasal oxygen insufflation, routine use of oxygen supplementation is recommended.

Scrapie of sheep and chronic wasting disease (CWD) of cervids are transmissible prion diseases. Milk and placenta have been identified as sources of scrapie prions but do not explain horizontal transmission. In contrast, CWD prions have been reported in saliva, urine and feces, which are thought to be responsible for horizontal transmission. While the titers of CWD prions have been measured in feces, levels in saliva or urine are unknown. Because sheep produce ~17 L/day of saliva, and scrapie prions are present in tongue and salivary glands of infected sheep, we asked if scrapie prions are shed in saliva. We inoculated transgenic (Tg) mice expressing ovine prion protein, Tg(OvPrP) mice, with saliva from seven Cheviot sheep with scrapie. Six of seven samples transmitted prions to Tg(OvPrP) mice with titers of -0.5 to 1.7 log ID₅₀ U/ml. Similarly, inoculation of saliva samples from two mule deer with CWD transmitted prions to Tg(ElkPrP) mice with titers of -1.1 to -0.4 log ID₅₀ U/ml. Assuming similar shedding kinetics for salivary prions as those for fecal prions of deer, we estimated the secreted salivary prion dose over a 10-mo period to be as high as 8.4 log ID₅₀ units for sheep and 7.0 log ID₅₀ units for deer. These estimates are similar to 7.9 log ID₅₀ units of fecal CWD prions for deer. Because saliva is mostly swallowed, salivary prions may reinfect tissues of the gastrointestinal tract and contribute to fecal prion shedding. Salivary prions shed into the environment provide an additional mechanism for horizontal prion transmission.