A Quantitative Assessment of the Amount of Prion Diverted to Category 1 
Materials and Wastewater During Processing 
Amie Adkin1,*, Neil Donaldson1, Louise Kelly1,2Article first published 
online: 24 DEC 2012
DOI: 10.1111/j.1539-6924.2012.01922.x
© 2012 Society for Risk Analysis 
 Keywords:Abattoir;bovine spongiform encephalopathy;QRA;scrapie;TSE
In this article the development and parameterization of a quantitative 
assessment is described that estimates the amount of TSE infectivity that is 
present in a whole animal carcass (bovine spongiform encephalopathy [BSE] for 
cattle and classical/atypical scrapie for sheep and lambs) and the amounts that 
subsequently fall to the floor during processing at facilities that handle 
specified risk material (SRM). BSE in cattle was found to contain the most oral 
doses, with a mean of 9864 BO ID50s (310, 38840) in a whole carcass compared to 
a mean of 1851 OO ID50s (600, 4070) and 614 OO ID50s (155, 1509) for a sheep 
infected with classical and atypical scrapie, respectively. Lambs contained the 
least infectivity with a mean of 251 OO ID50s (83, 548) for classical scrapie 
and 1 OO ID50s (0.2, 2) for atypical scrapie. The highest amounts of infectivity 
falling to the floor and entering the drains from slaughtering a whole carcass 
at SRM facilities were found to be from cattle infected with BSE at rendering 
and large incineration facilities with 7.4 BO ID50s (0.1, 29), intermediate 
plants and small incinerators with a mean of 4.5 BO ID50s (0.1, 18), and 
collection centers, 3.6 BO ID50s (0.1, 14). The lowest amounts entering drains 
are from lambs infected with classical and atypical scrapie at intermediate 
plants and atypical scrapie at collection centers with a mean of 3 × 10−7 OO 
ID50s (2 × 10−8, 1 × 10−6) per carcass. The results of this model provide key 
inputs for the model in the companion paper published here. 
it would have been interesting if CWD would have been included in this 
study. ...tss 
Saturday, December 15, 2012 
Bovine spongiform encephalopathy: the effect of oral exposure dose on 
attack rate and incubation period in cattle -- an update 5 December 2012
Detection of Protease-Resistant Prion Protein in Water from a CWD-Endemic 
Area 
65 
Tracy A. Nichols*1,2, Bruce Pulford1, Christy Wyckoff1,2, Crystal 
Meyerett1, Brady Michel1, Kevin Gertig3, Jean E. Jewell4, Glenn C. Telling5 and 
M.D. Zabel1 1Department of Microbiology, Immunology and Pathology, College of 
Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort 
Collins, CO 80523, USA 2National Wildlife Research Center, Wildlife Services, 
United States Department of Agriculture, Fort Collins, Colorado, 80521, USA 
3Fort Collins Water and Treatment Operations, Fort Collins, Colorado, 80521, USA 
4 Department of Veterinary Sciences, Wyoming State Veterinary Laboratory, 
University of Wyoming, Laramie, Wyoming, 82070, USA 5Department of Microbiology, 
Immunology, Molecular Genetics and Neurology, Sanders Brown Center on Aging, 
University of Kentucky, Lexington, Kentucky, 40536, USA * Corresponding author- 
tracy.a.nichols@aphis.usda.gov 
Chronic wasting disease (CWD) is the only known transmissible spongiform 
encephalopathy affecting free-ranging wildlife. Experimental and epidemiological 
data indicate that CWD can be transmitted horizontally and via blood and saliva, 
although the exact mode of natural transmission remains unknown. Substantial 
evidence suggests that prions can persist in the environment, implicating it as 
a potential prion reservoir and transmission vehicle. CWD- positive animals can 
contribute to environmental prion load via biological materials including 
saliva, blood, urine and feces, shedding several times their body weight in 
possibly infectious excreta in their lifetime, as well as through decomposing 
carcasses. Sensitivity limitations of conventional assays hamper evaluation of 
environmental prion loads in water. Here we show the ability of serial protein 
misfolding cyclic amplification (sPMCA) to amplify minute amounts of CWD prions 
in spiked water samples at a 1:1 x106 , and protease-resistant prions in 
environmental and municipal-processing water samples from a CWD endemic area. 
Detection of CWD prions correlated with increased total organic carbon in water 
runoff from melting winter snowpack. These data suggest prolonged persistence 
and accumulation of prions in the environment that may promote CWD transmission. 
snip... 
The data presented here demonstrate that sPMCA can detect low levels of 
PrPCWD in the environment, corroborate previous biological and experimental data 
suggesting long term persistence of prions in the environment2,3 and imply that 
PrPCWD accumulation over time may contribute to transmission of CWD in areas 
where it has been endemic for decades. This work demonstrates the utility of 
sPMCA to evaluate other environmental water sources for PrPCWD, including 
smaller bodies of water such as vernal pools and wallows, where large numbers of 
cervids congregate and into which prions from infected animals may be shed and 
concentrated to infectious levels. 
snip...end...full text at ; 
PLoS ONE. 2008; 3(8): e2969. 
Prion Infected Meat-and-Bone Meal Is Still Infectious after Biodiesel 
Production 
The epidemic of bovine spongiform encephalopathy (BSE) has led to a 
world-wide drop in the market for beef by-products, such as Meat-and-Bone Meal 
(MBM), a fat-containing but mainly proteinaceaous product traditionally used as 
an animal feed supplement. While normal rendering is insufficient, the 
production of biodiesel from MBM has been suggested to destroy infectivity from 
transmissible spongiform encephalopathies (TSEs). In addition to producing fuel, 
this method simultaneously generates a nutritious solid residue. In our study we 
produced biodiesel from MBM under defined conditions using a modified form of 
alkaline methanolysis. We evaluated the presence of prion in the three resulting 
phases of the biodiesel reaction (Biodiesel, Glycerol and Solid Residue) in 
vitro and in vivo. Analysis of the reaction products from 263K scrapie infected 
MBM led to no detectable immunoreactivity by Western Blot. Importantly, and in 
contrast to the biochemical results the solid MBM residue from the reaction 
retained infectivity when tested in an animal bioassay. Histochemical analysis 
of hamster brains inoculated with the solid residue showed typical spongiform 
degeneration and vacuolation. Re-inoculation of these brains into a new cohort 
of hamsters led to onset of clinical scrapie symptoms within 75 days, suggesting 
that the specific infectivity of the prion protein was not changed during the 
biodiesel process. The biodiesel reaction cannot be considered a viable prion 
decontamination method for MBM, although we observed increased survival time of 
hamsters and reduced infectivity greater than 6 log orders in the solid MBM 
residue. Furthermore, results from our study compare for the first time prion 
detection by Western Blot versus an infectivity bioassay for analysis of 
biodiesel reaction products. We could show that biochemical analysis alone is 
insufficient for detection of prion infectivity after a biodiesel process. 
New studies on the heat resistance of hamster-adapted scrapie agent: 
Threshold survival after ashing at 600°C suggests an inorganic template of 
replication 
Abstract 
One-gram samples from a pool of crude brain tissue from hamsters infected 
with the 263K strain of hamster-adapted scrapie agent were placed in covered 
quartz-glass crucibles and exposed for either 5 or 15 min to dry heat at 
temperatures ranging from 150°C to 1,000°C. Residual infectivity in the treated 
samples was assayed by the intracerebral inoculation of dilution series into 
healthy weanling hamsters, which were observed for 10 months; disease 
transmissions were verified by Western blot testing for proteinase-resistant 
protein in brains from clinically positive hamsters. Unheated control tissue 
contained 9.9 log10LD50/g tissue; after exposure to 150°C, titers equaled or 
exceeded 6 log10LD50/g, and after exposure to 300°C, titers equaled or exceeded 
4 log10LD50/g. Exposure to 600°C completely ashed the brain samples, which, when 
reconstituted with saline to their original weights, transmitted disease to 5 of 
35 inoculated hamsters. No transmissions occurred after exposure to 
1,000°C.
see full text: 
Friday, December 14, 2012 
Susceptibility of domestic cats to chronic wasting disease 
Friday, December 14, 2012 
Susceptibility Chronic Wasting Disease (CWD) in wild cervids to Humans 2005 
- December 14, 2012 
Friday, December 14, 2012 
DEFRA U.K. What is the risk of Chronic Wasting Disease CWD being introduced 
into Great Britain? A Qualitative Risk Assessment October 2012 
Tuesday, December 18, 2012 
A Growing Threat How deer breeding could put public trust wildlife at risk 
Wednesday, March 28, 2012 
VARIABLY PROTEASE-SENSITVE PRIONOPATHY IS TRANSMISSIBLE, price of prion 
poker goes up again $ 
Tuesday, December 18, 2012
Bioassay Studies Support the Potential for Iatrogenic Transmission of 
Variant Creutzfeldt Jakob Disease through Dental Procedures 
Wednesday, May 16, 2012 
Alzheimer’s disease and Transmissible Spongiform Encephalopathy prion 
disease, Iatrogenic, what if ? 
Proposal ID: 29403 
RIP MOM 12/14/97 confirmed hvCJD...TSS December 25, 2012
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