Immediate and ongoing detection of prions in the blood of hamsters and deer 
following oral, nasal, or blood inoculations
Alan M. Elder1, Davin M. Henderson1, Amy V. Nalls1, Edward A. Hoover1, 
Anthony E. Kincaid2,3, Jason C. Bartz2 and Candace K. Mathiason1# 
+ Author Affiliations 1Department of Microbiology, Immunology and 
Pathology, Colorado State University, Fort Collins, CO, 80523, United States of 
America 2Medical Microbiology and Immunology, Creighton University, Omaha, NE, 
68178, United States of America 3Department of Pharmacy Sciences, Creighton 
University, Omaha, NE, 68178, United States of America 
ABSTRACT
Infectious prions traverse epithelial barriers to gain access to the 
circulatory system, yet the temporal parameters of transepithelial transport and 
persistence in the blood over time remains unknown. We used wbRT-QuIC to analyze 
whole blood collected from TSE-inoculated deer and hamsters throughout the 
entire incubation period for the presence of PrPC-conversion competent amyloid 
(PrPC-CCA). We observed PrPC-CCA in the blood of TSE-inoculated hosts throughout 
disease course from minutes post exposure to terminal disease. 
 FOOTNOTES
↵#To whom any correspondence should be addressed: 
candace.mathiason@colostate.edu, 1619 Campus Delivery, Fort Collins, Co 
80523-1619, 970 491-3975 Copyright © 2015, American Society for Microbiology. 
All Rights Reserved. 
Prions circulate in the blood of prion-infected hosts, including humans 
with variant Creutzfeldt- Jakob disease. Determining the parameters of 
blood-borne prions during the long asymptomatic phase of disease characteristic 
of all prion diseases has been a long-standing problem in prion biology. Elder 
et. al (p. 7421–7424) have demonstrated amyloid formation, a biomarker for 
prions, in the blood of prion-infected rodent and cervid hosts as early as 15 
minutes post-mucosal or -intravenous infection. This prionemia persists 
throughout the disease course, indicating a role for hematogenous prions 
throughout the preclinical stage of illness. 
snip... 
UPDATED June 20, 2015 further into this study ;
***This is the first report of the detection of PrPC-CCA in the blood of 
animals within minutes of exposure to TSE inoculum. The use of PrPC-CCA to 
detect prions in tissues and bodily fluids of TSE-infected hosts has been shown 
to be as sensitive as a bioassay and has provided insight into a variety of 
prion diseases (15–18). The immediate detection of the point-source inoculum was 
seen following exposure to mucosal surfaces in the nose and gut; the same 
immediate detection of PrPC-CCA was seen following i.v. injection. This 
immediate detection indicates that the mucosae of the nasal cavity and the gut 
provide an inefficient anatomical barrier to prion entry—i.e., exposure to the 
mucosal surfaces does not differ significantly, in terms of temporal systemic 
spread, from injection directly into blood. These data, along with reports from 
Kincaid et al. (19) and Jeffrey et al. (20), demonstrate that mucosal surfaces 
are not capable of preventing the near-immediate passage of the inoculum into 
underlying lamina propria and blood. The absence of an efficient mucosal barrier 
to prion infection has significant implications for the pathogenesis of prion 
diseases. *** In addition, there are experimental and safety concerns, including 
the consideration that the blood of any animal exposed to prions may immediately 
contain prions. 
I don’t have access to the full text of this study, thus, not knowing 
exactly which TSE this study was speaking of ‘’We observed PrPC-CCA in the blood 
of TSE-inoculated hosts throughout disease course from minutes post exposure to 
terminal disease.’’ , but seems concerning to me. ...kind regards, terry
*** The potential impact of prion diseases on human health was greatly 
magnified by the recognition that interspecies transfer of BSE to humans by beef 
ingestion resulted in vCJD. While changes in animal feed constituents and 
slaughter practices appear to have curtailed vCJD, there is concern that CWD of 
free-ranging deer and elk in the U.S. might also cross the species barrier. 
Thus, consuming venison could be a source of human prion disease. Whether BSE 
and CWD represent interspecies scrapie transfer or are newly arisen prion 
diseases is unknown. Therefore, the possibility of transmission of prion disease 
through other food animals cannot be ruled out. There is evidence that vCJD can 
be transmitted through blood transfusion. There is likely a pool of unknown size 
of asymptomatic individuals infected with vCJD, and there may be asymptomatic 
individuals infected with the CWD equivalent. These circumstances represent a 
potential threat to blood, blood products, and plasma supplies. 
Circulation of prions within dust on a scrapie affected farm
Kevin C Gough1, Claire A Baker2, Hugh A Simmons3, Steve A Hawkins3 and Ben 
C Maddison2*
Abstract
Prion diseases are fatal neurological disorders that affect humans and 
animals. Scrapie of sheep/goats and Chronic Wasting Disease (CWD) of deer/elk 
are contagious prion diseases where environmental reservoirs have a direct link 
to the transmission of disease. Using protein misfolding cyclic amplification we 
demonstrate that scrapie PrPSc can be detected within circulating dusts that are 
present on a farm that is naturally contaminated with sheep scrapie. The 
presence of infectious scrapie within airborne dusts may represent a possible 
route of infection and illustrates the difficulties that may be associated with 
the effective decontamination of such scrapie affected premises. 
snip... 
Discussion
We present biochemical data illustrating the airborne movement of scrapie 
containing material within a contaminated farm environment. We were able to 
detect scrapie PrPSc within extracts from dusts collected over a 70 day period, 
in the absence of any sheep activity. We were also able to detect scrapie PrPSc 
within dusts collected within pasture at 30 m but not at 60 m distance away from 
the scrapie contaminated buildings, suggesting that the chance of contamination 
of pasture by scrapie contaminated dusts decreases with distance from 
contaminated farm buildings. PrPSc amplification by sPMCA has been shown to 
correlate with infectivity and amplified products have been shown to be 
infectious [14,15]. These experiments illustrate the potential for low dose 
scrapie infectivity to be present within such samples. We estimate low ng levels 
of scrapie positive brain equivalent were deposited per m2 over 70 days, in a 
barn previously occupied by sheep affected with scrapie. This movement of dusts 
and the accumulation of low levels of scrapie infectivity within this 
environment may in part explain previous observations where despite stringent 
pen decontamination regimens healthy lambs still became scrapie infected after 
apparent exposure from their environment alone [16]. The presence of sPMCA 
seeding activity and by inference, infectious prions within dusts, and their 
potential for airborne dissemination is highly novel and may have implications 
for the spread of scrapie within infected premises. The low level circulation 
and accumulation of scrapie prion containing dust material within the farm 
environment will likely impede the efficient decontamination of such scrapie 
contaminated buildings unless all possible reservoirs of dust are removed. 
Scrapie containing dusts could possibly infect animals during feeding and 
drinking, and respiratory and conjunctival routes may also be involved. It has 
been demonstrated that scrapie can be efficiently transmitted via the nasal 
route in sheep [17], as is also the case for CWD in both murine models and in 
white tailed deer [18-20]. 
The sources of dust borne prions are unknown but it seems reasonable to 
assume that faecal, urine, skin, parturient material and saliva-derived prions 
may contribute to this mobile environmental reservoir of infectivity. This work 
highlights a possible transmission route for scrapie within the farm 
environment, and this is likely to be paralleled in CWD which shows strong 
similarities with scrapie in terms of prion dissemination and disease 
transmission. The data indicate that the presence of scrapie prions in dust is 
likely to make the control of these diseases a considerable challenge. 
Wednesday, April 22, 2015 
Circulation of prions within dust on a scrapie affected farm 
Saturday, March 15, 2014 
Potential role of soil properties in the spread of CWD in western Canada 
Sunday, November 3, 2013 
Environmental Impact Statements; Availability, etc.: Animal Carcass 
Management [Docket No. APHIS-2013-0044] 
Friday, February 08, 2013 
*** Behavior of Prions in the Environment: Implications for Prion Biology 
Sunday, September 01, 2013 
hunting over gut piles and CWD TSE prion disease 
*** Spraker suggested an interesting explanation for the occurrence of CWD. 
The deer pens at the Foot Hills Campus were built some 30-40 years ago by a Dr. 
Bob Davis. At or abut that time, allegedly, some scrapie work was conducted at 
this site. When deer were introduced to the pens they occupied ground that had 
previously been occupied by sheep. ...
also, see where even decades back, the USDA had the same thought as they do 
today with CWD, not their problem...see page 27 below as well, where USDA stated 
back then, the same thing they stated in the state of Pennsylvania, not their 
damn business, once they escape, and they said the same thing about CWD in 
general back then ; 
”The occurrence of CWD must be viewed against the contest of the locations 
in which it occurred. It was an incidental and unwelcome complication of the 
respective wildlife research programmes. Despite it’s subsequent recognition as 
a new disease of cervids, therefore justifying direct investigation, no specific 
research funding was forthcoming. The USDA veiwed it as a wildlife problem and 
consequently not their province!” ...page 26. 
”The occurrence of CWD must be viewed against the contest of the locations 
in which it occurred. It was an incidental and unwelcome complication of the 
respective wildlife research programmes. Despite it’s subsequent recognition as 
a new disease of cervids, therefore justifying direct investigation, no specific 
research funding was forthcoming. The USDA veiwed it as a wildlife problem and 
consequently not their province!” ...page 26. 
sound familiar $$$ 
Sunday, January 06, 2013 
USDA TO PGC ONCE CAPTIVES ESCAPE 
*** "it‘s no longer its business.” 
Sunday, October 27, 2013 
A Kiss of a Prion: New Implications for Oral Transmissibility 
***please read this*** 
98 | Veterinary Record | January 24, 2015
EDITORIAL
Scrapie: a particularly persistent pathogen
Cristina Acín
Resistant prions in the environment have been the sword of Damocles for 
scrapie control and eradication. Attempts to establish which physical and 
chemical agents could be applied to inactivate or moderate scrapie infectivity 
were initiated in the 1960s and 1970s,with the first study of this type focusing 
on the effect of heat treatment in reducing prion infectivity (Hunter and 
Millson 1964). Nowadays, most of the chemical procedures that aim to inactivate 
the prion protein are based on the method developed by Kimberlin and 
collaborators (1983). This procedure consists of treatment with 20,000 parts per 
million free chlorine solution, for a minimum of one hour, of all surfaces that 
need to be sterilised (in laboratories, lambing pens, slaughterhouses, and so 
on). Despite this, veterinarians and farmers may still ask a range of questions, 
such as ‘Is there an official procedure published somewhere?’ and ‘Is there an 
international organisation which recommends and defines the exact method of 
scrapie decontamination that must be applied?’
From a European perspective, it is difficult to find a treatment that could 
be applied, especially in relation to the disinfection of surfaces in lambing 
pens of affected flocks. A 999/2001 EU regulation on controlling spongiform 
encephalopathies (European Parliament and Council 2001) did not specify a 
particular decontamination measure to be used when an outbreak of scrapie is 
diagnosed. There is only a brief recommendation in Annex VII concerning the 
control and eradication of transmissible spongiform encephalopathies (TSE 
s).
Chapter B of the regulation explains the measures that must be applied if 
new caprine animals are to be introduced to a holding where a scrapie outbreak 
has previously been diagnosed. In that case, the statement indicates that 
caprine animals can be introduced ‘provided that a cleaning and disinfection of 
all animal housing on the premises has been carried out following 
destocking’.
Issues around cleaning and disinfection are common in prion prevention 
recommendations, but relevant authorities, veterinarians and farmers may have 
difficulties in finding the specific protocol which applies. The European Food 
and Safety Authority (EFSA ) published a detailed report about the efficacy of 
certain biocides, such as sodium hydroxide, sodium hypochlorite, guanidine and 
even a formulation of copper or iron metal ions in combination with hydrogen 
peroxide, against prions (EFSA 2009). The report was based on scientific 
evidence (Fichet and others 2004, Lemmer and others 2004, Gao and others 2006, 
Solassol and others 2006) but unfortunately the decontamination measures were 
not assessed under outbreak conditions.
The EFSA Panel on Biological Hazards recently published its conclusions on 
the scrapie situation in the EU after 10 years of monitoring and control of the 
disease in sheep and goats (EFSA 2014), and one of the most interesting findings 
was the Icelandic experience regarding the effect of disinfection in scrapie 
control. The Icelandic plan consisted of: culling scrapie-affected sheep or the 
whole flock in newly diagnosed outbreaks; deep cleaning and disinfection of 
stables, sheds, barns and equipment with high pressure washing followed by 
cleaning with 500 parts per million of hypochlorite; drying and treatment with 
300 ppm of iodophor; and restocking was not permitted for at least two years. 
Even when all of these measures were implemented, scrapie recurred on several 
farms, indicating that the infectious agent survived for years in the 
environment, even as many as 16 years after restocking (Georgsson and others 
2006).
In the rest of the countries considered in the EFSA (2014) report, 
recommendations for disinfection measures were not specifically defined at the 
government level. In the report, the only recommendation that is made for sheep 
is repopulation with sheep with scrapie-resistant genotypes. This reduces the 
risk of scrapie recurrence but it is difficult to know its effect on the 
infection.
Until the EFSA was established (in May 2003), scientific opinions about TSE 
s were provided by the Scientific Steering Committee (SSC) of the EC, whose 
advice regarding inactivation procedures focused on treating animal waste at 
high temperatures (150°C for three hours) and high pressure alkaline hydrolysis 
(SSC 2003). At the same time, the TSE Risk Management Subgroup of the Advisory 
Committee on Dangerous Pathogens (ACDP) in the UK published guidance on safe 
working and the prevention of TSE infection. Annex C of the ACDP report 
established that sodium hypochlorite was considered to be effective, but only if 
20,000 ppm of available chlorine was present for at least one hour, which has 
practical limitations such as the release of chlorine gas, corrosion, 
incompatibility with formaldehyde, alcohols and acids, rapid inactivation of its 
active chemicals and the stability of dilutions (ACDP 2009).
In an international context, the World Organisation for Animal Health (OIE) 
does not recommend a specific disinfection protocol for prion agents in its 
Terrestrial Code or Manual. Chapter 4.13 of the Terrestrial Code, General 
recommendations on disinfection and disinsection (OIE 2014), focuses on 
foot-and-mouth disease virus, mycobacteria and Bacillus anthracis, but not on 
prion disinfection. Nevertheless, the last update published by the OIE on bovine 
spongiform encephalopathy (OIE 2012) indicates that few effective 
decontamination techniques are available to inactivate the agent on surfaces, 
and recommends the removal of all organic material and the use of sodium 
hydroxide, or a sodium hypochlorite solution containing 2 per cent available 
chlorine, for more than one hour at 20ºC.
The World Health Organization outlines guidelines for the control of TSE s, 
and also emphasises the importance of mechanically cleaning surfaces before 
disinfection with sodium hydroxide or sodium hypochlorite for one hour (WHO 
1999).
Finally, the relevant agencies in both Canada and the USA suggest that the 
best treatments for surfaces potentially contaminated with prions are sodium 
hydroxide or sodium hypochlorite at 20,000 ppm. This is a 2 per cent solution, 
while most commercial household bleaches contain 5.25 per cent sodium 
hypochlorite. It is therefore recommended to dilute one part 5.25 per cent 
bleach with 1.5 parts water (CDC 2009, Canadian Food Inspection Agency 
2013).
So what should we do about disinfection against prions? First, it is 
suggested that a single protocol be created by international authorities to 
homogenise inactivation procedures and enable their application in all 
scrapie-affected countries. Sodium hypochlorite with 20,000 ppm of available 
chlorine seems to be the procedure used in most countries, as noted in a paper 
summarised on p 99 of this issue of Veterinary Record (Hawkins and others 2015). 
But are we totally sure of its effectiveness as a preventive measure in a 
scrapie outbreak? Would an in-depth study of the recurrence of scrapie disease 
be needed?
What we can conclude is that, if we want to fight prion diseases, and 
specifically classical scrapie, we must focus on the accuracy of diagnosis, 
monitoring and surveillance; appropriate animal identification and control of 
movements; and, in the end, have homogeneous and suitable protocols to 
decontaminate and disinfect lambing barns, sheds and equipment available to 
veterinarians and farmers. Finally, further investigations into the resistance 
of prion proteins in the diversity of environmental surfaces are required.
References
snip...
98 | Veterinary Record | January 24, 2015
Persistence of ovine scrapie infectivity in a farm environment following 
cleaning and decontamination 
Steve A. C. Hawkins, MIBiol, Pathology Department1, Hugh A. Simmons, BVSc 
MRCVS, MBA, MA Animal Services Unit1, Kevin C. Gough, BSc, PhD2 and Ben C. 
Maddison, BSc, PhD3 + Author Affiliations
1Animal and Plant Health Agency, Woodham Lane, New Haw, Addlestone, Surrey 
KT15 3NB, UK 2School of Veterinary Medicine and Science, The University of 
Nottingham, Sutton Bonington, Loughborough, Leicestershire LE12 5RD, UK 3ADAS 
UK, School of Veterinary Medicine and Science, The University of Nottingham, 
Sutton Bonington, Loughborough, Leicestershire LE12 5RD, UK E-mail for 
correspondence: ben.maddison@adas.co.uk Abstract Scrapie of sheep/goats and 
chronic wasting disease of deer/elk are contagious prion diseases where 
environmental reservoirs are directly implicated in the transmission of disease. 
In this study, the effectiveness of recommended scrapie farm decontamination 
regimens was evaluated by a sheep bioassay using buildings naturally 
contaminated with scrapie. Pens within a farm building were treated with either 
20,000 parts per million free chorine solution for one hour or were treated with 
the same but were followed by painting and full re-galvanisation or replacement 
of metalwork within the pen. Scrapie susceptible lambs of the PRNP genotype 
VRQ/VRQ were reared within these pens and their scrapie status was monitored by 
recto-anal mucosa-associated lymphoid tissue. All animals became infected over 
an 18-month period, even in the pen that had been subject to the most stringent 
decontamination process. These data suggest that recommended current guidelines 
for the decontamination of farm buildings following outbreaks of scrapie do 
little to reduce the titre of infectious scrapie material and that environmental 
recontamination could also be an issue associated with these premises. 
SNIP...
Discussion
Thorough pressure washing of a pen had no effect on the amount of 
bioavailable scrapie infectivity (pen B). The routine removal of prions from 
surfaces within a laboratory setting is treatment for a minimum of one hour with 
20,000 ppm free chlorine, a method originally based on the use of brain 
macerates from infected rodents to evaluate the effectiveness of decontamination 
(Kimberlin and others 1983). Further studies have also investigated the 
effectiveness of hypochlorite disinfection of metal surfaces to simulate the 
decontamination of surgical devices within a hospital setting. Such treatments 
with hypochlorite solution were able to reduce infectivity by 5.5 logs to lower 
than the sensitivity of the bioassay used (Lemmer and others 2004). Analogous 
treatment of the pen surfaces did not effectively remove the levels of scrapie 
infectivity over that of the control pens, indicating that this method of 
decontamination is not effective within a farm setting. This may be due to the 
high level of biological matrix that is present upon surfaces within the farm 
environment, which may reduce the amount of free chlorine available to 
inactivate any infectious prion. Remarkably 1/5 sheep introduced into pen D had 
also became scrapie positive within nine months, with all animals in this pen 
being RAMALT positive by 18 months of age. Pen D was no further away from the 
control pen (pen A) than any of the other pens within this barn. Localised hot 
spots of infectivity may be present within scrapie-contaminated environments, 
but it is unlikely that pen D area had an amount of scrapie contamination that 
was significantly different than the other areas within this building. 
Similarly, there were no differences in how the biosecurity of pen D was 
maintained, or how this pen was ventilated compared with the other pens. This 
observation, perhaps, indicates the slower kinetics of disease uptake within 
this pen and is consistent with a more thorough prion removal and 
recontamination. These observations may also account for the presence of 
inadvertent scrapie cases within other studies, where despite stringent 
biosecurity, control animals have become scrapie positive during challenge 
studies using barns that also housed scrapie-affected animals (Ryder and others 
2009). The bioassay data indicate that the exposure of the sheep to a farm 
environment after decontamination efforts thought to be effective in removing 
scrapie is sufficient for the animals to become infected with scrapie. The main 
exposure routes within this scenario are likely to be via the oral route, during 
feeding and drinking, and respiratory and conjunctival routes. It has been 
demonstrated that scrapie infectivity can be efficiently transmitted via the 
nasal route in sheep (Hamir and others 2008), as is the case for CWD in both 
murine models and in white-tailed deer (Denkers and others 2010, 2013). 
Recently, it has also been demonstrated that CWD prions presented as dust when 
bound to the soil mineral montmorillonite can be infectious via the nasal route 
(Nichols and others 2013). When considering pens C and D, the actual source of 
the infectious agent in the pens is not known, it is possible that biologically 
relevant levels of prion survive on surfaces during the decontamination regimen 
(pen C). With the use of galvanising and painting (pen D) covering and sealing 
the surface of the pen, it is possible that scrapie material recontaminated the 
pens by the movement of infectious prions contained within dusts originating 
from other parts of the barn that were not decontaminated or from other areas of 
the farm.
Given that scrapie prions are widespread on the surfaces of affected farms 
(Maddison and others 2010a), irrespective of the source of the infectious prions 
in the pens, this study clearly highlights the difficulties that are faced with 
the effective removal of environmentally associated scrapie infectivity. This is 
likely to be paralleled in CWD which shows strong similarities to scrapie in 
terms of both the dissemination of prions into the environment and the facile 
mode of disease transmission. These data further contribute to the understanding 
that prion diseases can be highly transmissible between susceptible individuals 
not just by direct contact but through highly stable environmental reservoirs 
that are refractory to decontamination.
The presence of these environmentally associated prions in farm buildings 
make the control of these diseases a considerable challenge, especially in 
animal species such as goats where there is lack of genetic resistance to 
scrapie and, therefore, no scope to re-stock farms with animals that are 
resistant to scrapie.
Scrapie Sheep Goats Transmissible spongiform encephalopathies (TSE) 
Accepted October 12, 2014. Published Online First 31 October 2014 
Monday, November 3, 2014 
Persistence of ovine scrapie infectivity in a farm environment following 
cleaning and decontamination
PPo3-22:
Detection of Environmentally Associated PrPSc on a Farm with Endemic 
Scrapie
Ben C. Maddison,1 Claire A. Baker,1 Helen C. Rees,1 Linda A. Terry,2 Leigh 
Thorne,2 Susan J. Belworthy2 and Kevin C. Gough3 1ADAS-UK LTD; Department of 
Biology; University of Leicester; Leicester, UK; 2Veterinary Laboratories 
Agency; Surry, KT UK; 3Department of Veterinary Medicine and Science; University 
of Nottingham; Sutton Bonington, Loughborough UK
Key words: scrapie, evironmental persistence, sPMCA
Ovine scrapie shows considerable horizontal transmission, yet the routes of 
transmission and specifically the role of fomites in transmission remain poorly 
defined. Here we present biochemical data demonstrating that on a 
scrapie-affected sheep farm, scrapie prion contamination is widespread. It was 
anticipated at the outset that if prions contaminate the environment that they 
would be there at extremely low levels, as such the most sensitive method 
available for the detection of PrPSc, serial Protein Misfolding Cyclic 
Amplification (sPMCA), was used in this study. We investigated the distribution 
of environmental scrapie prions by applying ovine sPMCA to samples taken from a 
range of surfaces that were accessible to animals and could be collected by use 
of a wetted foam swab. Prion was amplified by sPMCA from a number of these 
environmental swab samples including those taken from metal, plastic and wooden 
surfaces, both in the indoor and outdoor environment. At the time of sampling 
there had been no sheep contact with these areas for at least 20 days prior to 
sampling indicating that prions persist for at least this duration in the 
environment. These data implicate inanimate objects as environmental reservoirs 
of prion infectivity which are likely to contribute to disease transmission. 
2012 
PO-039: A comparison of scrapie and chronic wasting disease in white-tailed 
deer 
Justin Greenlee, Jodi Smith, Eric Nicholson US Dept. Agriculture; 
Agricultural Research Service, National Animal Disease Center; Ames, IA USA 
snip...
The results of this study suggest that there are many similarities in the 
manifestation of CWD and scrapie in WTD after IC inoculation including early and 
widespread presence of PrPSc in lymphoid tissues, clinical signs of depression 
and weight loss progressing to wasting, and an incubation time of 21-23 months. 
Moreover, western blots (WB) done on brain material from the obex region have a 
molecular profile similar to CWD and distinct from tissues of the cerebrum or 
the scrapie inoculum. However, results of microscopic and IHC examination 
indicate that there are differences between the lesions expected in CWD and 
those that occur in deer with scrapie: amyloid plaques were not noted in any 
sections of brain examined from these deer and the pattern of immunoreactivity 
by IHC was diffuse rather than plaque-like. 
*** After a natural route of exposure, 100% of WTD were susceptible to 
scrapie. 
Deer developed clinical signs of wasting and mental depression and were 
necropsied from 28 to 33 months PI. Tissues from these deer were positive for 
PrPSc by IHC and WB. Similar to IC inoculated deer, samples from these deer 
exhibited two different molecular profiles: samples from obex resembled CWD 
whereas those from cerebrum were similar to the original scrapie inoculum. On 
further examination by WB using a panel of antibodies, the tissues from deer 
with scrapie exhibit properties differing from tissues either from sheep with 
scrapie or WTD with CWD. Samples from WTD with CWD or sheep with scrapie are 
strongly immunoreactive when probed with mAb P4, however, samples from WTD with 
scrapie are only weakly immunoreactive. In contrast, when probed with mAb’s 6H4 
or SAF 84, samples from sheep with scrapie and WTD with CWD are weakly 
immunoreactive and samples from WTD with scrapie are strongly positive. This 
work demonstrates that WTD are highly susceptible to sheep scrapie, but on first 
passage, scrapie in WTD is differentiable from CWD. 
2011 
*** After a natural route of exposure, 100% of white-tailed deer were 
susceptible to scrapie. 
Thursday, March 26, 2015 
Increased Infectivity of Anchorless Mouse Scrapie Prions in Transgenic Mice 
Overexpressing Human Prion Protein 
Sunday, March 29, 2015 
Uncommon prion disease induced in macaque ten years after scrapie 
inoculation 
Tuesday, December 16, 2014 
Evidence for zoonotic potential of ovine scrapie prions 
Hervé Cassard,1, n1 Juan-Maria Torres,2, n1 Caroline Lacroux,1, Jean-Yves 
Douet,1, Sylvie L. Benestad,3, Frédéric Lantier,4, Séverine Lugan,1, Isabelle 
Lantier,4, Pierrette Costes,1, Naima Aron,1, Fabienne Reine,5, Laetitia 
Herzog,5, Juan-Carlos Espinosa,2, Vincent Beringue5, & Olivier Andréoletti1, 
Affiliations Contributions Corresponding author Journal name: Nature 
Communications Volume: 5, Article number: 5821 DOI: doi:10.1038/ncomms6821 
Received 07 August 2014 Accepted 10 November 2014 Published 16 December 2014 
Article tools Citation Reprints Rights & permissions Article metrics 
Abstract 
Although Bovine Spongiform Encephalopathy (BSE) is the cause of variant 
Creutzfeldt Jakob disease (vCJD) in humans, the zoonotic potential of scrapie 
prions remains unknown. Mice genetically engineered to overexpress the human 
prion protein (tgHu) have emerged as highly relevant models for gauging the 
capacity of prions to transmit to humans. These models can propagate human 
prions without any apparent transmission barrier and have been used used to 
confirm the zoonotic ability of BSE. Here we show that a panel of sheep scrapie 
prions transmit to several tgHu mice models with an efficiency comparable to 
that of cattle BSE. The serial transmission of different scrapie isolates in 
these mice led to the propagation of prions that are phenotypically identical to 
those causing sporadic CJD (sCJD) in humans. These results demonstrate that 
scrapie prions have a zoonotic potential and raise new questions about the 
possible link between animal and human prions.
Subject terms: Biological sciences• Medical research At a glance
why do we not want to do TSE transmission studies on chimpanzees $ 
5. A positive result from a chimpanzee challenged severly would likely 
create alarm in some circles even if the result could not be interpreted for 
man. I have a view that all these agents could be transmitted provided a large 
enough dose by appropriate routes was given and the animals kept long enough. 
Until the mechanisms of the species barrier are more clearly understood it might 
be best to retain that hypothesis. 
snip... 
R. BRADLEY 
Friday, January 30, 2015
Scrapie: a particularly persistent pathogen
Wednesday, March 18, 2015 
Chronic Wasting Disease CWD Confirmed Texas Trans Pecos March 18, 
2015
Wednesday, March 25, 2015 
Chronic Wasting Disease CWD Cases Confirmed In New Mexico 2013 and 2014 
UPDATE 2015
Tuesday, December 20, 2011
CHRONIC WASTING DISEASE CWD WISCONSIN Almond Deer (Buckhorn Flats) 
FarmUpdate DECEMBER 2011The CWD infection rate was nearly 80%, the highest ever 
in a North American captive herd. RECOMMENDATION: That the Board approve the 
purchase of 80acres of land for $465,000 for the Statewide Wildlife Habitat 
Program inPortage County and approve the restrictions on public use of the 
site.SUMMARY:
For Immediate Release Thursday, October 2, 2014 
Dustin Vande Hoef 515/281-3375 or 515/326-1616 (cell) or 
Dustin.VandeHoef@IowaAgriculture.gov
TEST RESULTS FROM CAPTIVE DEER HERD WITH CHRONIC WASTING DISEASE RELEASED 
79.8 percent of the deer tested positive for the disease
DES MOINES – The Iowa Department of Agriculture and Land Stewardship today 
announced that the test results from the depopulation of a quarantined captive 
deer herd in north-central Iowa showed that 284 of the 356 deer, or 79.8% of the 
herd, tested positive for Chronic Wasting Disease (CWD). The owners of the 
quarantined herd have entered into a fence maintenance agreement with the Iowa 
Department of Agriculture and Land Stewardship,which requires the owners to 
maintain the 8’ foot perimeter fence around the herd premises for five years 
after the depopulation was complete and the premises had been cleaned and 
disinfected CWD is a progressive, fatal, degenerative neurological disease of 
farmed and free-ranging deer, elk, and moose. There is no known treatment or 
vaccine for CWD. CWD is not a disease that affects humans.On July 18, 2012, USDA 
Animal and Plant Health Inspection Service’s (APHIS)National Veterinary Services 
Lab in Ames, IA confirmed that a male whitetail deer harvested from a hunting 
preserve in southeast IA was positive for CWD. An investigation revealed that 
this animal had just been introduced into the hunting preserve from the 
above-referenced captive deer herd in north-central Iowa.The captive deer herd 
was immediately quarantined to prevent the spread of CWD. The herd has remained 
in quarantine until its depopulation on August 25 to 27, 2014.The Iowa 
Department of Agriculture and Land Stewardship participated in a joint operation 
to depopulate the infected herd with USDA Veterinary Services, which was the 
lead agency, and USDA Wildlife Services.Federal indemnity funding became 
available in 2014. USDA APHIS appraised the captive deer herd of 376 animals at 
that time, which was before depopulation and testing, at $1,354,250. At that 
time a herd plan was developed with the owners and officials from USDA and the 
Iowa Department of Agriculture and Land Stewardship.Once the depopulation was 
complete and the premises had been cleaned and disinfected, indemnity of 
$917,100.00 from the USDA has been or will be paid to the owners as compensation 
for the 356 captive deer depopulated.The Iowa Department of Agriculture and Land 
Stewardship operates a voluntary CWD program for farms that sell live animals. 
Currently 145 Iowa farms participate in the voluntary program. The 
above-referenced captive deer facility left the voluntary CWD program prior to 
the discovery of the disease as they had stopped selling live animals. All deer 
harvested in a hunting preserve must be tested for CWD. -30-
*** see history of this CWD blunder here ; 
On June 5, 2013, DNR conducted a fence inspection, after gaining approval 
from surrounding landowners, and confirmed that the fenced had beencut or 
removed in at least four separate locations; that the fence had degraded and was 
failing to maintain the enclosure around the Quarantined Premises in at least 
one area; that at least three gates had been opened;and that deer tracks were 
visible in and around one of the open areas in the sand on both sides of the 
fence, evidencing movement of deer into the Quarantined Premises.
Tuesday, October 07, 2014 
*** Wisconsin white-tailed deer tested positive for CWD on a Richland 
County breeding farm, and a case of CWD has been discovered on a Marathon County 
hunting preserve 
*** Wisconsin 16 age limit on testing dead deer Game Farm CWD Testing 
Protocol Needs To Be Revised 
Approximately 4,200 fawns, defined as deer under 1 year of age, were 
sampled from the eradication zone over the last year. The majority of fawns 
sampled were between the ages of 5 to 9 months, though some were as young as 1 
month. 
*** Two of the six fawns with CWD detected were 5 to 6 months old. 
All six of the positive fawns were taken from the core area of the CWD 
eradication zone where the highest numbers of positive deer have been 
identified. ... 
snip...
"Finding CWD prions in both lymph and brain tissues of deer this young is 
slightly surprising," said Langenberg, "and provides information that CWD 
infection and illness may progress more rapidly in a white-tailed deer than 
previously suspected. Published literature suggests that CWD doesn't cause 
illness in a deer until approximately 16 months of age. Our fawn data shows that 
a few wild white-tailed deer may become sick from CWD or may transmit the 
disease before they reach that age of 16 months." ... see full text and more 
here ; Saturday, February 04, 2012 
Wisconsin 16 MONTH age limit on testing dead deer Game Farm CWD Testing 
Protocol Needs To Be Revised 
Thursday, July 03, 2014 
*** How Chronic Wasting Disease is affecting deer population and what’s the 
risk to humans and pets? 
Tuesday, July 01, 2014 
*** CHRONIC WASTING DISEASE CWD TSE PRION DISEASE, GAME FARMS, AND 
POTENTIAL RISK FACTORS THERE FROM 
=========================================== 
spreading cwd around...
Between 1996 and 2002, chronic wasting disease was diagnosed in 39 herds of 
farmed elk in Saskatchewan in a single epidemic. All of these herds were 
depopulated as part of the Canadian Food Inspection Agency’s (CFIA) disease 
eradication program. Animals, primarily over 12 mo of age, were tested for the 
presence CWD prions following euthanasia. Twenty-one of the herds were linked 
through movements of live animals with latent CWD from a single infected source 
herd in Saskatchewan, 17 through movements of animals from 7 of the secondarily 
infected herds. 
***The source herd is believed to have become infected via importation of 
animals from a game farm in South Dakota where CWD was subsequently diagnosed 
(7,4). A wide range in herd prevalence of CWD at the time of herd depopulation 
of these herds was observed. Within-herd transmission was observed on some 
farms, while the disease remained confined to the introduced animals on other 
farms. 
spreading cwd around... 
Friday, May 13, 2011 
Chronic Wasting Disease (CWD) outbreaks and surveillance program in the 
Republic of Korea 
Hyun-Joo Sohn, Yoon-Hee Lee, Min-jeong Kim, Eun-Im Yun, Hyo-Jin Kim, 
Won-Yong Lee, Dong-Seob Tark, In- Soo Cho, Foreign Animal Disease Research 
Division, National Veterinary Research and Quarantine Service, Republic of Korea 
Chronic wasting disease (CWD) has been recognized as an important prion 
disease in native North America deer and Rocky mountain elks. The disease is a 
unique member of the transmissible spongiform encephalopathies (TSEs), which 
naturally affects only a few species. CWD had been limited to USA and Canada 
until 2000. 
On 28 December 2000, information from the Canadian government showed that a 
total of 95 elk had been exported from farms with CWD to Korea. These consisted 
of 23 elk in 1994 originating from the so-called “source farm” in Canada, and 72 
elk in 1997, which had been held in pre export quarantine at the “source 
farm”.Based on export information of CWD suspected elk from Canada to Korea, CWD 
surveillance program was initiated by the Ministry of Agriculture and Forestry 
(MAF) in 2001. 
All elks imported in 1997 were traced back, however elks imported in 1994 
were impossible to identify. CWD control measures included stamping out of all 
animals in the affected farm, and thorough cleaning and disinfection of the 
premises. In addition, nationwide clinical surveillance of Korean native 
cervids, and improved measures to ensure reporting of CWD suspect cases were 
implemented. 
Total of 9 elks were found to be affected. CWD was designated as a 
notifiable disease under the Act for Prevention of Livestock Epidemics in 2002. 
Additional CWD cases - 12 elks and 2 elks - were diagnosed in 2004 and 
2005. 
Since February of 2005, when slaughtered elks were found to be positive, 
all slaughtered cervid for human consumption at abattoirs were designated as 
target of the CWD surveillance program. Currently, CWD laboratory testing is 
only conducted by National Reference Laboratory on CWD, which is the Foreign 
Animal Disease Division (FADD) of National Veterinary Research and Quarantine 
Service (NVRQS). 
In July 2010, one out of 3 elks from Farm 1 which were slaughtered for the 
human consumption was confirmed as positive. Consequently, all cervid – 54 elks, 
41 Sika deer and 5 Albino deer – were culled and one elk was found to be 
positive. Epidemiological investigations were conducted by Veterinary 
Epidemiology Division (VED) of NVRQS in collaboration with provincial veterinary 
services. 
Epidemiologically related farms were found as 3 farms and all cervid at 
these farms were culled and subjected to CWD diagnosis. Three elks and 5 
crossbreeds (Red deer and Sika deer) were confirmed as positive at farm 2. 
All cervids at Farm 3 and Farm 4 – 15 elks and 47 elks – were culled and 
confirmed as negative. 
Further epidemiological investigations showed that these CWD outbreaks were 
linked to the importation of elks from Canada in 1994 based on circumstantial 
evidences. 
In December 2010, one elk was confirmed as positive at Farm 5. 
Consequently, all cervid – 3 elks, 11 Manchurian Sika deer and 20 Sika deer – 
were culled and one Manchurian Sika deer and seven Sika deer were found to be 
positive. This is the first report of CWD in these sub-species of deer. 
Epidemiological investigations found that the owner of the Farm 2 in CWD 
outbreaks in July 2010 had co-owned the Farm 5. 
In addition, it was newly revealed that one positive elk was introduced 
from Farm 6 of Jinju-si Gyeongsang Namdo. All cervid – 19 elks, 15 crossbreed 
(species unknown) and 64 Sika deer – of Farm 6 were culled, but all confirmed as 
negative. 
”The occurrence of CWD must be viewed against the contest of the locations 
in which it occurred. It was an incidental and unwelcome complication of the 
respective wildlife research programmes. Despite it’s subsequent recognition as 
a new disease of cervids, therefore justifying direct investigation, no specific 
research funding was forthcoming. The USDA veiwed it as a wildlife problem and 
consequently not their province!” ...page 26. 
Sunday, January 06, 2013 
USDA TO PGC ONCE CAPTIVES ESCAPE 
*** "it‘s no longer its business.” 
Sunday, July 13, 2014 
Louisiana deer mystery unleashes litigation 6 does still missing from CWD 
index herd in Pennsylvania Great Escape 
Saturday, June 29, 2013 
PENNSYLVANIA CAPTIVE CWD INDEX HERD MATE YELLOW *47 STILL RUNNING LOOSE IN 
INDIANA, YELLOW NUMBER 2 STILL MISSING, AND OTHERS ON THE RUN STILL IN LOUISIANA 
Tuesday, June 11, 2013 
*** CWD GONE WILD, More cervid escapees from more shooting pens on the 
loose in Pennsylvania 
Wednesday, September 04, 2013 
***cwd - cervid captive livestock escapes, loose and on the run in the 
wild... 
Friday, January 30, 2015
*** Scrapie: a particularly persistent pathogen ***
Tuesday, January 06, 2015 
APHIS Provides Additional Information on Chronic Wasting Disease (CWD) 
Indemnity Requests January 5, 2015 05:26 PM EST
31 Jan 2015 at 20:14 GMT 
*** Singeltary reply ; 
ruminant feed ban for cervids in the United States ? 
31 Jan 2015 at 20:14 GMT 
Friday, April 17, 2015 
Assessing Transmissible Spongiform Encephalopathy Species Barriers with an 
In Vitro Prion Protein Conversion Assay 
>>> show some preliminary results suggesting that bobcats (Lynx 
rufus) may be susceptible to white-tailed deer (Odocoileus virginianus) chronic 
wasting disease agent. 
Monday, March 09, 2015 
Chronic Wasting Disease CWD TSE prion and human animal risk factor there 
from 
Sunday, April 12, 2015 
*** Research Project: Transmission, Differentiation, and Pathobiology of 
Transmissible Spongiform Encephalopathies 2014 Annual Report *** 
 http://transmissiblespongiformencephalopathy.blogspot.com/2015/04/research-project-transmission.html 
Wednesday, April 15, 2015 
KURU Transmissible Spongiform Encephalopthy TSE Prion Disease 
TSS

