Salivary prions in sheep and deer

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.Key words: scrapie, chronic wasting disease, saliva, horizontal transmission, titers     

Intra-host mathematical model of chronic wasting disease dynamics in deer (Odocoileus)

Bioassays of native cervid hosts have established the presence of infectious chronic wasting disease (CWD) prions in saliva, blood, urine, and feces of clinically diseased and pre-clinical infected deer. The intra-host trafficking of prions from the time of initial infection to shedding has been less well defined. We created a discrete-time compartmentalized model to simulate the misfolding catalysis, trafficking, and shedding of infectious prions throughout the organ systems of CWD-infected cervids. Using parameter values derived from experimental infections of North American deer (Odocoileus spp.), the exponential-based model predicts prion deposition over time with: 1) nervous tissues containing the highest deposition of prions at 20 months post-infection and 2) excreted fluids containing low levels of prions throughout infection with the highest numbers of prions predicted to be shed in saliva and feces (as high as 10 lethal doses (1.34 × 10²⁹ prions) in 11–15 months). These findings are comparable to prion deposition described in literature as assayed by conventional and ultrasensitive amplification assays. The comparison of our model to published data suggests that highly sensitive assays (sPMCA, RT-QuIC, and bioassay) are appropriate for early prion detection in bodily fluids and secretions. The model provides a view of intra-host prion catalysis leading to pre-clinical shedding and provides a framework for continued development of antemortem diagnostic methods.     

Early detection of chronic wasting disease prions in urine of pre-symptomatic deer by real-time quaking-induced conversion assay

Chronic wasting disease (CWD) is a prion disease of captive and free-ranging deer (Odocoileus spp), elk (Cervus elaphus nelsonii) and moose (Alces alces shirasi). Unlike in most other prion diseases, in CWD prions are shed in urine and feces, which most likely contributes to the horizontal transmission within and between cervid species. To date, CWD ante-mortem diagnosis is only possible by immunohistochemical detection of protease resistant prion protein (PrP^Sc) in tonsil or recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsies, which requires anesthesia of animals. We report on detection of CWD prions in urine collected from pre-symptomatic deer and in fecal extracts by using real time quaking-induced conversion (RT-QuIC). This assay can be useful for non-invasive pre-symptomatic diagnosis and surveillance of CWD.     

Two different scrapie prions isolated in Japanese sheep flocks

Two different scrapie prion strains with different characteristics were obtained from two sheep naturally infected with scrapie in Japan. In mice transmission, one (Tsukuba-1) showed shorter incubation periods (133+/-2 days) than the other (Tsukuba-2) (288+/-5 days). Spongiform changes and accumulation of an abnormal isoform of prion protein (PrP(Sc)) were observed throughout the brain in Tsukuba-1 inoculated mice, while the lesions and the PrP(Sc) accumulation were localized in the brain stem of mice with Tsukuba-2. Western blot analysis showed that there was no strain-specific glycoform of PrP(Sc) within these two strains. A super-infection experiment revealed that neither strain interfered with the other's propagation.     

Cross-species transmission of CWD prions

Prions cause fatal neurodegenerative diseases in humans and animals and can be transmitted zoonotically. Chronic wasting disease (CWD) is a highly transmissible prion disease of wild deer and elk that affects cervids over extensive regions of the United States and Canada. The risk of cross-species CWD transmission has been experimentally evaluated in a wide array of mammals, including non-human primates and mouse models expressing human cellular prion protein. Here we review the determinants of cross-species CWD transmission, and propose a model that may explain a structural barrier for CWD transmission to humans.     

Tissue-specific biochemical differences between chronic wasting disease prions isolated from free-ranging white-tailed deer (Odocoileus virginianus)

Chronic wasting disease (CWD) is an invariably fatal prion disease affecting cervid species worldwide. Prions can manifest as distinct strains that can influence disease pathology and transmission. CWD is profoundly lymphotropic, and most infected cervids likely shed peripheral prions replicated in lymphoid organs. However, CWD is a neurodegenerative disease, and most research on prion strains has focused on neurogenic prions. Thus, a knowledge gap exists comparing neurogenic prions to lymphogenic prions. In this study, we compared prions from the obex and lymph nodes of naturally exposed white-tailed deer to identify potential biochemical strain differences. Here, we report biochemical evidence of strain differences between the brain and lymph node from these animals. Conformational stability assays, glycoform ratio analyses, and immunoreactivity scanning across the structured domain of the prion protein that refolds into the amyloid aggregate of the infectious prion reveal significantly more structural and glycoform variation in lymphogenic prions than neurogenic prions. Surprisingly, we observed greater biochemical differences among neurogenic prions than lymphogenic prions across individuals. We propose that the lymphoreticular system propagates a diverse array of prions from which the brain selects a more restricted pool of prions that may be quite different than those from another individual of the same species. Future work should examine the biological and zoonotic impact of these biochemical differences and examine more cervids from multiple locations to determine if these differences are conserved across species and locations.     

CWD prions remain infectious after passage through the digestive system of coyotes (Canis latrans)

ABSTRACT

Chronic wasting disease (CWD) is a geographically expanding prion disease of wild and captive cervids in North America. Disease can be transmitted directly, animal to animal, or indirectly via the environment. CWD contamination can occur residually in the environment via soil, water, and forage following deposition of bodily fluids such as urine, saliva, and feces, or by the decomposition of carcasses. Recent work has indicated that plants may even take up prions into the stems and leaves. When a carcass or gut pile is present in the environment, a large number of avian and mammalian species visit and consume the carrion. Additionally, predators like coyotes, likely select for disease-compromised cervids. Natural cross-species CWD transmission has not been documented, however, passage of infectious prion material has been observed in the feces of crows. In this study we evaluated the ability of CWD-infected brain material to pass through the gastrointestinal tract of coyotes (Canis latrans) following oral ingestion, and be infectious in a cervidized transgenic mouse model. Results from this study indicate that coyotes can pass infectious prions via their feces for at least 3 days post ingestion, demonstrating that mammalian scavengers could contribute to the translocation and contamination of CWD in the environment.     

Interaction among deer in a chronic wasting disease endemic zone

Although it is known that chronic wasting disease (CWD) can be transmitted by both direct animal-to-animal contact and contact with contaminated environments, the relative role of each mechanism in the spread of CWD in free-ranging populations has yet to be defined. We investigated patterns of interaction between mule deer (Odocoileus hemionus) in order to understand how factors such as season and landscape may influence patterns of disease spread in these populations. Using location data from male and female Global Positioning System (GPS)-collared mule deer in 5 study areas located in and around a CWD-endemic zone in southern Saskatchewan, Canada, we quantified close proximity events, or events involving both spatial and temporal overlap of individuals. We defined close proximity events as occurrences in which 2 deer were located <25 m apart at the same point in time. We looked at seasonal variation in the probability of close proximity events, as well as landscape factors associated with these events when compared to areas of shared space use, or spatial overlap alone. Overall probability of an individual GPS-collared deer being located in close proximity to another GPS-collared deer was 0.092 (n = 107). The early gestation (16 Dec–31 Mar) and late gestation (1 Apr–15 May) seasons had the highest probability of close proximity events occurring, and same-sex pairs were more likely to be found in close proximity than between-sex pairs during all seasons aside from the rut (1 Nov–15 Dec). High probability of close proximity events during the gestation seasons agrees with the tendency of mule deer to aggregate into large groups during late winter and suggests that this may be an important time period for CWD transmission to occur. Close proximity events occurred more in cropland and wetland than expected based on availability, whereas close proximity events occurred less than expected in grassland. The opposite was true for spatial overlap between individuals, which occurred more than expected in areas of low elevation and rugged terrain and in grassland or shrub–wood habitats. These results suggest that cropland may be a higher risk habitat for direct and indirect CWD transmission between individuals and that, although coulees and other areas of rugged topography are less likely to be associated with close proximity events, those areas may be more likely to contain environmental contamination in CWD-affected areas due to common use by multiple deer. © 2011 The Wildlife Society.     

A rare 300 kilometer dispersal by an adult male white‐tailed deer

Despite the key roles that dispersal plays in individual animal fitness and meta‐population gene flow, it remains one of the least understood behaviors in many species. In large mammalian herbivores, dispersals might span long distances and thereby influence landscape‐level ecological processes, such as infectious disease spread. Here, we describe and analyze an exceptional long‐distance dispersal by an adult white‐tailed deer (Odocoileus virginianus) in the central United States. We also conducted a literature survey to compare the dispersal to previous studies. This dispersal was remarkable for its length, duration, and the life history stage of the dispersing individual. Dispersal is typical of juvenile deer seeking to establish postnatal home ranges, but this dispersal was undertaken by an adult male (age = 3.5). This individual dispersed ~300 km over a 22‐day period by moving, on average, 13.6 km/day and achieving a straight‐line distance of ~215 km, which was ~174 km longer than any other distance recorded for an adult male deer in our literature survey. During the dispersal, which occurred during the hunting season, the individual crossed a major river seven times, an interstate highway, a railroad, and eight state highways. Movements during the dispersal were faster (mean = 568.1 m/h) and more directional than those during stationary home range periods before and after the dispersal (mean = 56.9 m/h). Likewise, movements during the dispersal were faster (mean = 847.8 m/h) and more directional at night than during the day (mean = 166.4 m/h), when the individual frequently sheltered in forest cover. This natural history event highlights the unpredictable nature of dispersal and has important implications for landscape‐level processes such as chronic wasting disease transmission in cervids. More broadly, our study underscores how integrating natural history observations with modern technology holds promise for understanding potentially high impact but rarely recorded ecological events.     

Wildlife disease elimination and density dependence

Disease control by managers is a crucial response to emerging wildlife epidemics, yet the means of control may be limited by the method of disease transmission. In particular, it is widely held that population reduction, while effective for controlling diseases that are subject to density-dependent (DD) transmission, is ineffective for controlling diseases that are subject to frequency-dependent (FD) transmission. We investigate control for horizontally transmitted diseases with FD transmission where the control is via culling or harvest that is non-selective with respect to infection and the population can compensate through DD recruitment or survival. Using a mathematical model, we show that culling or harvesting can eradicate the disease, even when transmission dynamics are FD. Eradication can be achieved under FD transmission when DD birth or recruitment induces compensatory growth of new, healthy individuals, which has the net effect of reducing disease prevalence by dilution. We also show that if harvest is used simultaneously with vaccination, and there is high enough transmission coefficient, application of both controls may be less efficient than vaccination alone. We illustrate the effects of these control approaches on disease prevalence for chronic wasting disease in deer where the disease is transmitted directly among deer and through the environment.     

Metals in obex and retropharyngeal lymph nodes of Illinois white-tailed deer and their variations associated with CWD status

ABSTRACT

Prion proteins (PrP^C) are cell membrane glycoproteins that can be found in many cell types, but specially in neurons. Many studies have suggested PrP^C‘s participation in metal transport and cellular protection against stress in the central nervous system (CNS). On the other hand PrP^Sc, the misfolded isoform of PrP^C and the pathogenic agent in transmissible spongiform encephalopathies (TSE), has been associated with brain metal dyshomeostasis in prion diseases. Thus, changes in metal concentration associated with protein misfolding and aggregation have been reported for human and animal prion diseases, as well as for other neurodegenerative disorders, such as Parkinson's and Alzheimer's disease. The use of metal concentrations in tissues as surrogate markers for early detection of TSEs has been suggested. Studies on the accumulation of metals in free-ranging white-tailed deer have not been conducted. This study established concentrations of copper, iron, manganese, and magnesium in 2 diagnostic tissues used for CWD testing (obex and retropharyngeal lymph nodes (RLN)). We compared these concentrations between tissues and in relation to CWD status. We established reference intervals (RIs) for these metals and explored their ability to discriminate between CWD-positive and CWD-negative animals. Our results indicate that independent of CWD status, white-tailed deer accumulate higher concentrations of Fe, Mn and Mg in RLN than in obex. White-tailed deer infected with CWD accumulated significantly lower concentrations of Mn and Fe than CWD-negative deer. These patterns differed from other species infected with prion diseases. Overlapping values between CWD positive and negative groups indicate that evaluation of these metals in obex and RLN may not be appropriate as a diagnostic tool for CWD infection in white-tailed deer. Because the CWD-negative deer were included in constructing the RIs, high specificities were expected and should be interpreted with caution. Due to the low sensitivity derived from the RIs, we do not recommend using metal concentrations for disease discrimination.