Wisconsin Farm-Raised Deer Farms and CWD

[flounder9]

Friday, February 03, 2012


Wisconsin Farm-Raised Deer Farms and CWD there from 2012 report Singeltary et al



chronic-wasting-disease.blogspot.com/2012/02/wisconsin-farm-raised-deer-farms-and.html




TSS

[flounder9]

Saturday, February 04, 2012


Wisconsin 16 age limit on testing dead deer Game Farm CWD Testing Protocol Needs To Be Revised


chronic-wasting-disease.blogspot.com/2012/02/wisconsin-16-age-limit-on-testing-dead.html



TSS

[tickman1961]

All this blogger is doing is spreading unfounded fear. Wisconsin did a great thing hiring James "Dr Deer" Kroll, he knows more about CWD than the blogger ever will.

"I have written in extensive detail about the known science of CWD versus the junk science of CWD. Unfortunately in this world gone mad where most bureaucrats have lost their souls, they have predictably embraced the junk science and mindlessly force policies based on presumptuous and totally unsupportable nonsense. The CWD scam is a bureaucrats wet dream." - Ted Nugent

[flounder9]

Feb 6, 2012 15:14:45 GMT -5 tickman1961 said:

All this blogger is doing is spreading unfounded fear. Wisconsin did a great thing hiring James "Dr Deer" Kroll, he knows more about CWD than the blogger ever will.

"I have written in extensive detail about the known science of CWD versus the junk science of CWD. Unfortunately in this world gone mad where most bureaucrats have lost their souls, they have predictably embraced the junk science and mindlessly force policies based on presumptuous and totally unsupportable nonsense. The CWD scam is a bureaucrats wet dream." - Ted Nugent

i kindly disagree with you and teddy. and i hate to bust your bubble, but the great white hunter (teddy), does not know everything, except his own ego. the science speaks for itself. and when one starts bragging about one knowing more than anyone else about TSE Prions, your in trouble. because nobody knows everything, especially about TSE prion disease. i have been working on the incident of CWD in Wisconsin game farms since around Christmas. wrote the Wisconsin DNR, asking for information on these CWD petri dishes. they put me off a time or two, enough time for Dr Bourie to put together some watered down story about it the night before he finally sent me the data i was looking for. take that with how every many grains of salt you wish Sir. to me, it smells. ...




wfbf.com/ag-newswire/bourie-and-deer-farmers-weather-the-cwd-storm/




i can assure you that Wisconsin has _weathered_ nothing when it comes to CWD. now with a new strain of CWD i.e. the 'Wisconsin CWD strain', the price of poker goes up. are you all in





Friday, February 03, 2012


Wisconsin Farm-Raised Deer Farms and CWD there from 2012 report Singeltary et al


chronic-wasting-disease.blogspot.com/2012/02/wisconsin-farm-raised-deer-farms-and.html





Saturday, February 04, 2012


Wisconsin 16 age limit on testing dead deer Game Farm CWD Testing Protocol Needs To Be Revised


chronic-wasting-disease.blogspot.com/2012/02/wisconsin-16-age-limit-on-testing-dead.html




P35

ADAPTATION OF CHRONIC WASTING DISEASE (CWD) INTO HAMSTERS, EVIDENCE OF A WISCONSIN STRAIN OF CWD

Chad Johnson1, Judd Aiken2,3,4 and Debbie McKenzie4,5 1 Department of Comparative Biosciences, University of Wisconsin, Madison WI, USA 53706 2 Department of Agriculture, Food and Nutritional Sciences, 3 Alberta Veterinary Research Institute, 4.Center for Prions and Protein Folding Diseases, 5 Department of Biological Sciences, University of Alberta, Edmonton AB, Canada T6G 2P5

The identification and characterization of prion strains is increasingly important for the diagnosis and biological definition of these infectious pathogens. Although well-established in scrapie and, more recently, in BSE, comparatively little is known about the possibility of prion strains in chronic wasting disease (CWD), a disease affecting free ranging and captive cervids, primarily in North America. We have identified prion protein variants in the white-tailed deer population and demonstrated that Prnp genotype affects the susceptibility/disease progression of white-tailed deer to CWD agent. The existence of cervid prion protein variants raises the likelihood of distinct CWD strains. Small rodent models are a useful means of identifying prion strains. We intracerebrally inoculated hamsters with brain homogenates and phosphotungstate concentrated preparations from CWD positive hunter-harvested (Wisconsin CWD endemic area) and experimentally infected deer of known Prnp genotypes. These transmission studies resulted in clinical presentation in primary passage of concentrated CWD prions. Subclinical infection was established with the other primary passages based on the detection of PrPCWD in the brains of hamsters and the successful disease transmission upon second passage. Second and third passage data, when compared to transmission studies using different CWD inocula (Raymond et al., 2007) indicate that the CWD agent present in the Wisconsin white-tailed deer population is different than the strain(s) present in elk, mule-deer and white-tailed deer from the western United States endemic region.


www.istitutoveneto.it/prion_09/Abstracts_09.pdf




PPo3-7:

Prion Transmission from Cervids to Humans is Strain-dependent

Qingzhong Kong, Shenghai Huang,*Fusong Chen, Michael Payne, Pierluigi Gambetti and Liuting Qing Department of Pathology; Case western Reserve University; Cleveland, OH USA *Current address: Nursing Informatics; Memorial Sloan-Kettering Cancer Center; New York, NY USA

Key words: CWD, strain, human transmission

Chronic wasting disease (CWD) is a widespread prion disease in cervids (deer and elk) in North America where significant human exposure to CWD is likely and zoonotic transmission of CWD is a concern. Current evidence indicates a strong barrier for transmission of the classical CWD strain to humans with the PrP-129MM genotype. A few recent reports suggest the presence of two or more CWD strains. What remain unknown is whether individuals with the PrP-129VV/MV genotypes are also resistant to the classical CWD strain and whether humans are resistant to all natural or adapted cervid prion strains. Here we report that a human prion strain that had adopted the cervid prion protein (PrP) sequence through passage in cervidized transgenic mice efficiently infected transgenic mice expressing human PrP, indicating that the species barrier from cervid to humans is prion strain-dependent and humans can be vulnerable to novel cervid prion strains. Preliminary results on CWD transmission in transgenic mice expressing human PrP-129V will also be discussed.

Acknowledgement Supported by NINDS NS052319 and NIA AG14359.


PPo2-27:

Generation of a Novel form of Human PrPSc by Inter-species Transmission of Cervid Prions

Marcelo A. Barria,1 Glenn C. Telling,2 Pierluigi Gambetti,3 James A. Mastrianni4 and Claudio Soto1 1Mitchell Center for Alzheimer's disease and related Brain disorders; Dept of Neurology; University of Texas Houston Medical School; Houston, TX USA; 2Dept of Microbiology, Immunology & Molecular Genetics and Neurology; Sanders Brown Center on Aging; University of Kentucky Medical Center; Lexington, KY USA; 3Institute of Pathology; Case western Reserve University; Cleveland, OH USA; 4Dept of Neurology; University of Chicago; Chicago, IL USA

Prion diseases are infectious neurodegenerative disorders affecting humans and animals that result from the conversion of normal prion protein (PrPC) into the misfolded and infectious prion (PrPSc). Chronic wasting disease (CWD) of cervids is a prion disorder of increasing prevalence within the United States that affects a large population of wild and captive deer and elk. CWD is highly contagious and its origin, mechanism of transmission and exact prevalence are currently unclear. The risk of transmission of CWD to humans is unknown. Defining that risk is of utmost importance, considering that people have been infected by animal prions, resulting in new fatal diseases. To study the possibility that human PrPC can be converted into the infectious form by CWD PrPSc we performed experiments using the Protein Misfolding Cyclic Amplification (PMCA) technique, which mimic in vitro the process of prion replication. Our results show that cervid PrPSc can induce the pathological conversion of human PrPC, but only after the CWD prion strain has been stabilized by successive passages in vitro or in vivo. Interestingly, this newly generated human PrPSc exhibits a distinct biochemical pattern that differs from any of the currently known forms of human PrPSc, indicating that it corresponds to a novel human prion strain. Our findings suggest that CWD prions have the capability to infect humans, and that this ability depends on CWD strain adaptation, implying that the risk for human health progressively increases with the spread of CWD among cervids.



PPo2-7:

Biochemical and Biophysical Characterization of Different CWD Isolates

Martin L. Daus and Michael Beekes Robert Koch Institute; Berlin, Germany

Key words: CWD, strains, FT-IR, AFM

Chronic wasting disease (CWD) is one of three naturally occurring forms of prion disease. The other two are Creutzfeldt-Jakob disease in humans and scrapie in sheep. CWD is contagious and affects captive as well as free ranging cervids. As long as there is no definite answer of whether CWD can breach the species barrier to humans precautionary measures especially for the protection of consumers need to be considered. In principle, different strains of CWD may be associated with different risks of transmission to humans. Sophisticated strain differentiation as accomplished for other prion diseases has not yet been established for CWD. However, several different findings indicate that there exists more than one strain of CWD agent in cervids. We have analysed a set of CWD isolates from white-tailed deer and could detect at least two biochemically different forms of disease-associated prion protein PrPTSE. Limited proteolysis with different concentrations of proteinase K and/or after exposure of PrPTSE to different pH-values or concentrations of Guanidinium hydrochloride resulted in distinct isolate-specific digestion patterns. Our CWD isolates were also examined in protein misfolding cyclic amplification studies. This showed different conversion activities for those isolates that had displayed significantly different sensitivities to limited proteolysis by PK in the biochemical experiments described above. We further applied Fourier transform infrared spectroscopy in combination with atomic force microscopy. This confirmed structural differences in the PrPTSE of at least two disinct CWD isolates. The data presented here substantiate and expand previous reports on the existence of different CWD strains.


www.prion2010.org/bilder/prion_2010_program_latest_w_posters_4_.pdf?139&PHPSESSID=a30a38202cfec579000b77af81be3099




UPDATED DATA ON 2ND CWD STRAIN


Wednesday, September 08, 2010


CWD PRION CONGRESS SEPTEMBER 8-11 2010



chronic-wasting-disease.blogspot.com/2010/09/cwd-prion-2010.html





CJD9/10022

October 1994

Mr R.N. Elmhirst Chairman British Deer Farmers Association Holly Lodge Spencers Lane BerksWell Coventry CV7 7BZ

Dear Mr Elmhirst,

CREUTZFELDT-JAKOB DISEASE (CJD) SURVEILLANCE UNIT REPORT

Thank you for your recent letter concerning the publication of the third annual report from the CJD Surveillance Unit. I am sorry that you are dissatisfied with the way in which this report was published.

The Surveillance Unit is a completely independant outside body and the Department of Health is committed to publishing their reports as soon as they become available. In the circumstances it is not the practice to circulate the report for comment since the findings of the report would not be amended. In future we can ensure that the British Deer Farmers Association receives a copy of the report in advance of publication.

The Chief Medical Officer has undertaken to keep the public fully informed of the results of any research in respect of CJD. This report was entirely the work of the unit and was produced completely independantly of the the Department.

The statistical results reqarding the consumption of venison was put into perspective in the body of the report and was not mentioned at all in the press release. Media attention regarding this report was low key but gave a realistic presentation of the statistical findings of the Unit. This approach to publication was successful in that consumption of venison was highlighted only once by the media ie. in the News at one television proqramme.

I believe that a further statement about the report, or indeed statistical links between CJD and consumption of venison, would increase, and quite possibly give damaging credence, to the whole issue. From the low key media reports of which I am aware it seems unlikely that venison consumption will suffer adversely, if at all.


web.archive.org/web/20030511010117/http://www.bseinquiry.gov.uk/files/yb/1994/10/00003001.pdf




and why do we not want to do TSE transmission studies on chimpanzees $




snip...

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


collections.europarchive.org/tna/20080102222950/http://www.bseinquiry.gov.uk/files/yb/1990/09/23001001.pdf




Prions in Skeletal Muscles of Deer with Chronic Wasting Disease

Rachel C. Angers1,*, Shawn R. Browning1,*?, Tanya S. Seward2, Christina J. Sigurdson4,?, Michael W. Miller5, Edward A. Hoover4 and Glenn C. Telling1,2,3,§ + Author Affiliations

Abstract

The emergence of chronic wasting disease (CWD) in deer and elk in an increasingly wide geographic area, as well as the interspecies transmission of bovine spongiform encephalopathy to humans in the form of variant Creutzfeldt Jakob disease, have raised concerns about the zoonotic potential of CWD. Because meat consumption is the most likely means of exposure, it is important to determine whether skeletal muscle of diseased cervids contains prion infectivity. Here bioassays in transgenic mice expressing cervid prion protein revealed the presence of infectious prions in skeletal muscles of CWD-infected deer, demonstrating that humans consuming or handling meat from CWD-infected deer are at risk to prion exposure.

Received for publication 21 November 2005. Accepted for publication 13 January 2006.


www.sciencemag.org/cgi/content/abstract/sci;311/5764/1117




Journal of Virology, September 2009, p. 9608-9610, Vol. 83, No. 18 0022-538X/09/$08.00+0 doi:10.1128/JVI.01127-09 Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Prion Infectivity in Fat of Deer with Chronic Wasting Disease

Brent Race,# Kimberly Meade-White,# Richard Race, and Bruce Chesebro* Rocky Mountain Laboratories, 903 South 4th Street, Hamilton, Montana 59840

Received 2 June 2009/ Accepted 24 June 2009

ABSTRACT Top ABSTRACT TEXT REFERENCES

Chronic wasting disease (CWD) is a neurodegenerative prion disease of cervids. Some animal prion diseases, such as bovine spongiform encephalopathy, can infect humans; however, human susceptibility to CWD is unknown. In ruminants, prion infectivity is found in central nervous system and lymphoid tissues, with smaller amounts in intestine and muscle. In mice, prion infectivity was recently detected in fat. Since ruminant fat is consumed by humans and fed to animals, we determined infectivity titers in fat from two CWD-infected deer. Deer fat devoid of muscle contained low levels of CWD infectivity and might be a risk factor for prion infection of other species.

snip...

The highest risk of human contact with CWD might be through exposure to high-titer CNS tissue through accidental skin cuts or corneal contact at the time of harvest and butchering. However, the likelihood of a human consuming fat infected with a low titer of the CWD agent is much higher. It is impossible to remove all the fat present within muscle tissue, and fat consumption is inevitable when eating meat. Of additional concern is the fact that meat from an individual deer harvested by a hunter is typically consumed over multiple meals by the same group of people. These individuals would thus have multiple exposures to the CWD agent over time, which might increase the chance for transfer of infection.

In the Rocky Mountain region of North America, wild deer are subject to predation by wolves, coyotes, bears, and mountain lions. Although canines such as wolves and coyotes are not known to be susceptible to prion diseases, felines definitely are susceptible to BSE (9) and might also be infected by the CWD agent. Deer infected with the CWD agent are more likely to be killed by predators such as mountain lions (11). Peripheral tissues, including lymph nodes, muscle, and fat, which harbor prion infectivity are more accessible for consumption than CNS tissue, which has the highest level of infectivity late in disease. Therefore, infectivity in these peripheral tissues may be important in potential cross-species CWD transmissions in the wild.

The present finding of CWD infectivity in deer fat tissue raises the possibility that prion infectivity might also be found in fat tissue of other infected ruminants, such as sheep and cattle, whose fat and muscle tissues are more widely distributed in both the human and domestic-animal food chains. Although the infectivity in fat tissues is low compared to that in the CNS, there may be significant differences among species and between prion strains. Two fat samples from BSE agent-infected cattle were reported to be negative by bioassay in nontransgenic RIII mice (3, 6). However, RIII mice are 10,000-fold-less sensitive to BSE agent infection than transgenic mice expressing bovine PrP (4). It would be prudent to carry out additional infectivity assays on fat from BSE agent-infected cattle and scrapie agent-infected sheep using appropriate transgenic mice or homologous species to determine the risk from these sources.


jvi.asm.org/cgi/content/full/83/18/9608




stupid is, as stupid does, and some times you just can't fix stupid. ...




tss

[tickman1961]

CWD is not the death of whitetail deer hunting as we know it. Thanks for showing me which side you represent. Disease is part of mother natures plan, I am amazed at the sheer number of deer killed in the state of Wisconsin by hunters every year. As you are well aware, CWD was found in the state of Colorado in the 1960's and there are still thriving deer populations alive and well in that state. Obviously you have a hard on for those that disagree with you, claiming how much more you know and belittle others that take the opposite side. I appreciate your response and calling me out as stupid. Sounds like a typical hunter vs hunter agenda with disease being the main factor.

[hornharvester]

flounder,

Please make your point without the name calling. h.h.

[flounder9]

Feb 7, 2012 11:09:31 GMT -5 tickman1961 said:

CWD is not the death of whitetail deer hunting as we know it. Thanks for showing me which side you represent. Disease is part of mother natures plan, I am amazed at the sheer number of deer killed in the state of Wisconsin by hunters every year. As you are well aware, CWD was found in the state of Colorado in the 1960's and there are still thriving deer populations alive and well in that state. Obviously you have a hard on for those that disagree with you, claiming how much more you know and belittle others that take the opposite side. I appreciate your response and calling me out as stupid. Sounds like a typical hunter vs hunter agenda with disease being the main factor.

Sir, i don't know you, and did not call you out, or mean that comment ‘stupid is, as stupid does, and some times you can't fix stupid’ at the end of what I posted, as a direct statement to you as being stupid. i use that term in general, probably should not, but that's one of those stupid little quotes people use at the end of there comments, and i picked up that habit some times, when speaking of TSE prion regulations. my bad.

your comment 'typical hunter vs hunter agenda with disease being main factor'....

well Sir, you said a mouth full. and that is a disturbing factor with CWD. it should NOT be hunter against hunter, but that's what happens, as with baiting and feeding, natural baits vs piles of bait, etc. it's always hunter vs hunter. with fighting CWD, it seems the politics wins out over sound science. course that happens every time with human and animal TSE prion disease. updating hunters with the latest science on CWD should not be regarded as an attempt to split up hunters into different camps. seems when you intertwine business with politics and with sound science, the science gets buried. people have to interpret it as they can, but these same people need to have access to all the science, and that does not happen all the time. you call it junk science, because you do not agree with it, or it does not fit your agenda, and at the same time quote ted nugent as your source of sound science for cwd i.e. 'The CWD scam is a bureaucrats wet dream'. tell that to the 80% CWD infection rate of one game farm in Wisconsin, out of the 9 documented cwd infected game farms to date in Wisconsin, and that’s just the documented ones, and ponder what will happen if you continue down the same old road of allowing 500+ game farms in Wisconsin to continue to exist, or anywhere else, as far as i am concerned. but that does not matter of what i think, it's the science that others might not have been aware of, that matters, and in any future decision policy making. folks should at least have all the latest science to make sound decisions. at least that’s my opinion. my intentions are NOT, and never has been to split up the hunters. it’s just been to inform them of the rest of the science. I cannot stress enough, it’s not just about you, and what you consume as hunters. I don’t care what you eat. but once you expose yourself to the TSE Prion via CWD infected deer and or elk, and then go have medical procedures done, surgical, dental, donate blood, tissue, etc., then you expose everybody with the TSE Prion disease. then it becomes everybody’s business. ...



kind regards, terry

[tickman1961]

Bad apples exist in all walks of life. 9 out of 500 is approximately 2%, should we punish all because of the ignorance of the few? I understand your concern and welcome you to Hunting Indiana.

[Russ Koon]

I also appreciate your concern and your work in alerting everyone to the dangers and the progression of the spread of the disease. The truth isn't always what we WANT to hear, but it needs to be said and heard.

I had an uncle who was very fond of squirrel hunting, and always hunted them with a shotgun so he wouldn't destroy his favorite part, the brains, with a head shot from a rifle. His death happened years before I was aware of any suspected connection between the eating of squirrel brains and CJD.

His symptoms were consistent for the most part with CJD, but the diagnosis was the more commonly diagnosed "hardening of the arteries" causing early senile dementia and other related problems. He was seen only by local medical talent, no reason at the time to suspect anything more exotic than their diagnosis, and no sign of a cure for anything more exotic had it been suspected.

And no way of knowing for sure now, but I've always suspected CJD in his case, since hearing about it later. He was the only family member with the fondness for squirrel brains. Some of the rest of us who hunted with him tried them, but none of us liked them better than the meatier parts, so we were all content to let him have all the brains he desired form the entire bag.

And while I do agree with Ted's stance and enthusiasm on 2A issues and hunting, I also would agree with you on the wisdom of relying too heavily on his scientific expertise 8^).

[flounder9]

CWD TSE PRION UPDATE END OF YEAR DECEMBER 2013

Prion2013 Chronic Wasting Disease CWD risk factors, humans, domestic cats, blood, and mother to offspring transmission



HD.13: CWD infection in the spleen of humanized transgenic mice

Liuting Qing and Qingzhong Kong

Case Western Reserve University; Cleveland, OH USA

Chronic wasting disease (CWD) is a widespread prion disease in free-ranging and captive cervid species in North America, and there is evidence suggesting the existence of multiple CWD strains. The susceptibility of human CNS and peripheral organs to the various CWD prion strains remains largely unclear. Current literature suggests that the classical CWD strain is unlikely to infect human brain, but the potential for peripheral infection by CWD in humans is unknown. We detected protease-resistant PrpSc in the spleens of a few humanized transgenic mice that were intracerebrally inoculated with natural CWD isolates, but PrpSc was not detected in the brains of any of the CWD-inoculated mice. Our ongoing bioassays in humanized Tg mice indicate that intracerebral challenge with such PrpSc-positive humanized mouse spleen already led to prion disease in most animals.
*** These results indicate that the CWD prion may have the potential to infect human peripheral lymphoid tissues.


=====


HD.12: Comparative study of the distribution of the prion protein in the squirrel monkey (Saimiri sciureus) following experimental challenge with variant and sporadic CJD

Diane L. Ritchie,1 Paul Brown,2 Susan Gibson,3 Thomas R. Kreil,4 Christian Abee3 and James W. Ironside1

1National CJD Surveillance Unit; Edinburgh, UK; 2Bethesda; Bethesda, MD USA; 3Deparment of Comparative Medicine; University of South Alabama; Mobile, AL USA; 4Baxter Bioscience; Vienna, Austria

Introduction, Reports suggest that the number of tissues and organs showing the presence of the abnormal prion protein (PrPTSE) in variant CJD (vCJD) patients may be greater than previously thought. A limited peripheral involvement in some cases of sporadic CJD (sCJD) has also been reported. This accumulation of PrPTSE outside the brain has raised concerns about the possible iatrogenic transmission risk of vCJD. The squirrel monkey (Saimiri sciureus) has been shown to be highly susceptible to experimental challenge with human prion disease. Neuropathological and biochemical analyses of CNS tissue have shown that sCJD and vCJD can be distinguished in the squirrel monkey and that many of the strain characteristics that define these agents are conserved after transmission. Following on from these initial studies, immunohistochemistry and western blot analysis were performed on a wide range of peripheral tissues including, lymphoreticular tissues and peripheral neural tissue to establish the full-body distribution of PrPTSE in this primate animal model.

Materials and Methods. Brain homogenates from sCJD or vCJD patients were inoculated into the frontal cortex of squirrel monkeys. Animals were kept under constant clinical surveillance. At post-mortem, formalin fixed CNS tissue and a wide range of peripheral tissues were taken for immunohistochemical analysis together with frozen tissues taken for the biochemical detection of PrPTSE.

Results. Immunohistochemical analysis showed no evidence of PrPTSE deposition in peripheral tissues in either variant or sporadic CJD-infected animals. However, western blot assays detected PrPTSE in the spleen of a proportion of the vCJD- infected animals. The PrPTSE isotype resembled that detected in CNS tissue from the vCJD- infected animals and from human vCJD cases. ***In addition, western blot analysis detected PrPTSE in the spleen of a single animal following challenge with sporadic CJD. The PrPTSE type in this animal resembled that found in CNS tissue from the same animal, with a PrPTSE type similar to that found in human sCJD type 1 cases.

Conclusion. This study confirms the accumulation of PrPTSE in the CNS and spleen of a proportion of squirrel monkeys infected intra-cerebrally with human vCJD. Furthermore, this study extends the evidence that there may be a peripheral involvement in some cases of sCJD. PrPTSE typing confirms the conservation of PrPTSE type on transmission to the squirrel monkey and suggests that there are no tissue-specific adaptations in the biochemical phenotype of the agent strain following primate-to-primate transmission.


=====


Oral.15: Molecular barriers to zoonotic prion transmission: Comparison of the ability of sheep, cattle and deer prion disease isolates to convert normal human prion protein to its pathological isoform in a cell-free system

Marcelo A.Barria,1 Aru Balachandran,2 Masanori Morita,3 Tetsuyuki Kitamoto,4 Rona Barron,5 Jean Manson,5 Richard Kniqht,1 James W. lronside1 and Mark W. Head1

1National CJD Research and Surveillance Unit; Centre for Clinical Brain Sciences; School of Clinical Sciences; The University of Edinburgh; Edinburgh, UK; 2National and OIE Reference Laboratory for Scrapie and CWD; Canadian Food Inspection Agency; Ottawa Laboratory; Fallowfield. ON Canada; 3Infectious Pathogen Research Section; Central Research Laboratory; Japan Blood Products Organization; Kobe, Japan; 4Department of Neurological Science; Tohoku University Graduate School of Medicine; Sendai. Japan; 5Neurobiology Division; The Roslin Institute and R(D)SVS; University of Edinburgh; Easter Bush; Midlothian; Edinburgh, UK

Background. Bovine spongiform encephalopathy (BSE) is a known zoonotic prion disease, resulting in variant Creurzfeldt- Jakob disease (vCJD) in humans. In contrast, classical scrapie in sheep is thought to offer little or no danger to human health. However, a widening range of prion diseases have been recognized in cattle, sheep and deer. The risks posed by individual animal prion diseases to human health cannot be determined a priori and are difficult to assess empirically. The fundamemal event in prion disease pathogenesis is thought to be the seeded conversion of normal prion protein (PrPC) to its pathological isoform (PrPSc). Here we report the use of a rapid molecular conversion assay to test whether brain specimens from different animal prion diseases are capable of seeding the conversion of human PrPC ro PrPSc.

Material and Methods. Classical BSE (C-type BSE), H-type BSE, L-type BSE, classical scrapie, atypical scrapie, chronic wasting disease and vCJD brain homogenates were tested for their ability to seed conversion of human PrPC to PrPSc in protein misfolding cyclic amplification (PMCA) reactions. Newly formed human PrPSc was detected by protease digestion and western blotting using the antibody 3F4.

Results. C-type BSE and vCJD were found to efficiently convert PrPC to PrPSc. Scrapie failed to convert human PrPC to PrPSc. Of the other animal prion diseases tested only chronic wasting disease appeared to have the capability ro convert human PrPC to PrPSc. The results were consistent whether the human PrPC came from human brain, humanised transgenic mouse brain or from cultured human cells and the effect was more pronounced for PrPC with methionine at codon 129 compared with that with valine.

Conclusion. Our results show that none of the tested animal prion disease isolates are as efficient as C-type BSE and vCJD in converting human prion protein in this in vitro assay. *** However, they also show that there is no absolute barrier ro conversion of human prion protein in the case of chronic wasting disease.


=====


Invited.16: Studies of chronic wasting disease transmission in cervid and non-cervid species

Edward A, Hoover,1 Candace K. Mathiason,1 Davin M. Henderson,1 Nicholas J. Haley,1 Davis M. Seelig,1 Nathaniel D. Denkers,1 Amy V. Nalls,1 Mark D. Zabe,1 Glenn C. Telling,1 Fernando Goni2 and Thomas Wisniewski,2

1Prion Research Center; Colorado State University; Fort Collins, CO USA; 2New York University School of Medicine; New York, NY USA

How and why some misfolded proteins become horizontally transmitted agents and occasionally cross species barriers are issues fundamental to understanding prion disease. Chronic wasting disease (CWD) of cervids is perhaps a prototype of horizontal prion transmission, encompassing efficient mucosal uptake, lymphoid amplification, neuroinvasion, peripheralization, and dissemination via mucosal excretion. Efficient mucosal transmission of CWD in deer has been demonstrated by oral, nasal, aerosol, and indirect contact exposure. In addition, other studies (Mathiason CK, et al.) reported at the symposium support a significant role for pre- and/or postnatal transmission of CWD from doe to offspring. Accumulating, yet still incomplete, evidence also suggests that the period of relatively covert CWD infection may be longer than originally thought. Given the above, minimally invasive sensitive assays based on body fluids from live animals would aid substantially in understanding the biology of CWD. We have been applying seeded realtirne quaking-induced amplification of recombinant PrP substrates (i.e., RT-QuIC methodology) to:

(1) investigate antemortem CWD detection, and

(2) model PrP-based species barriers and trans-species adaptation-topics we previously explored using sPMCA and in vivo bioassays.

*** At this symposium, we report sensitive and specific detection CWD prions in saliva, urine, blood (Mathiason lab), and rectal and pharyngeal lymph node samples (Haley NJ, et al.) from pre-symptomatic and symptomatic experimentally and naturally exposed deer.

Other ongoing studies are employing RT-QuIC methodology to model amplification barriers among CWD, FSE, BSE, and CJD prions using cervine, feline, bovine, human, and promiscuous rPrP substrates and the above species prion seeds, cellular co-factors, and transgenic mice. Finally, in collaboration with the Wisniewski laboratory, we are conducting of experimental CWD vaccination studies in deer employing oral administration of an attenuated Salmonella vector expressing cervid PrP epitopes.


=====


AD.06: Detecting prions in the brain and blood of TSE-infected deer and hamsters

Alan Elder,1 Davin Henderson,1 Anca Selariu,1 Amy Nalls,1 Byron Caughey,2 Richard Bessen,1 Jason Bartz3 and Candace Mathiason1

1Colorado State University; Fort Collins, CO USA; 2NIH Rocky Mountain Laboratories; Hamilton, MT USA; 3Creighton University; Omaha, NE USA

While large quantities of protease resistant prion protein (PrPres) can be demonstrated by western blot or IHC in lymphoid biopsies or post-mortem brain tissues harvested from prion-infected animals, these conventional assays are less reliable as means to detect the small quantities of prions thought to be present in bodily fluids or associated with early and asymptomatic phases of TSE disease. The Real Time-Quaking Induced Conversion (RT-QuIC) assay is capable of detecting prions at concentrations below the level of sensitivity of conventional assays and provides a real-time fluorescent readout negating the use of proteases. We have made modifications to the RT-QuIC assay to utilize it for the detection of PrPres in brain and blood harvested from various species infected with prions. In this study, we analyzed CWD-infected deer and CWD/TME-infected hamster whole blood to determine the effect of:

(1) various anticoagulants,

(2) freezing and

(3) NaPTA precipitation.

Brain tissue and blood collected from naive deer and hamsters served as negative controls.

We were able to demonstrate amplifiable prions in

(1) brain and blood samples harvested from CWD/TME-infected animals,

(2) heparinized blood,

(3) frozen vs. fresh blood and

(4) NaPTA treated samples.

The RT-QuIC assay is able to detect PrPres in various species of animals and shows promise as an antemortem diagnostic tool for blood-borne TSEs.


=====


Oral.08: Mother to offspring transmission of chronic wasting disease in Reeve's Muntjac deer

Amy Nalls,1 Erin McNulty,1 Jenny Powers,2 Davis Seelig,1 Clare Hoover,1 Nicholas Haley,1 Jeanette Hayes-Klug,1 Kelly Anderson,1 Paula Stewart,3 Wilfred Goldmann,3 Edward A. Hoover1 and Candace K. Mathiason1

1Colorado State University; Fort Collins, CO USA; 2National Park Service; Fort Collins, CO USA; 3The Roslin Institute and Royal School of Veterinary Studies; Edinburgh, UK

To investigate the role mother to offspring transmission plays in chronic wasting disease (CWD), we have developed a cervid model employing the Reeve's muntjac deer (Muntiacus reevesi). Eight muntjac doe were orally inoculated with CWD and tested PrPCWD lymphoid positive by 4 mo post infection. Fourteen fawns were born to these eight CWD-infected doe-3 were born viable, 6 were born non-viable and 5 were harvested as fetuses from early or end-stage CWD-infected doe. All three viable fawns have demonstrated CWD IHC lymphoid biopsy positivity between 43 d post birth and 11 mo post birth. Two of these three CWD positive viable offspring have developed clinical signs consistent with TSE disease (28-33 mo post birth). Moreover, CWD prions have been detected by sPMCA in 11 of 16 tissues harvested from 6 full-term non-viable fawns and in 7 of 11 fetal tissues harvested in utero from the second and third trimester fetuses. Additional tissues and pregnancy related fluids from doe and offspring are being analyzed for CWD prions. *** In summary, using the muntjac deer model we have demonstrated CWD clinical disease in offspring born to CWD-infected doe, and in utero transmission of CWD from mother to offspring. These studies provide basis to further investigate the mechanisms of maternal transfer of prions.


=====


AD.63: Susceptibility of domestic cats to chronic wasting disease

Amy V.Nalls,1 Candace Mathiason,1 Davis Seelig,2 Susan Kraft,1 Kevin Carnes,1 Kelly Anderson,1 Jeanette Hayes-Klug1 and Edward A. Hoover1

1Colorado State University; Fort Collins, CO USA; 2University of Minnesota; Saint Paul, MN USA

Domestic and nondomestic cats have been shown to be susceptible to feline spongiform encephalopathy (FSE), almost certainly caused by consumption of bovine spongiform encephalopathy (BSE)-contaminated meat. Because domestic and free-ranging nondomestic felids scavenge cervid carcasses, including those in areas affected by chronic wasting disease (CWD), we evaluated the susceptibility of the domestic cat (Felis catus) to CWD infection experimentally. Cohorts of 5 cats each were inoculated either intracerebrally (IC) or orally (PO) with CWD-infected deer brain. At 40 and 42 mo post-inoculation, two IC-inoculated cats developed signs consistent with prion disease, including a stilted gait, weight loss, anorexia, polydipsia, patterned motor behaviors, head and tail tremors, and ataxia, and progressed to terminal disease within 5 mo. Brains from these two cats were pooled and inoculated into cohorts of cats by IC, PO, and intraperitoneal and subcutaneous (IP/SC) routes. Upon subpassage, feline-adapted CWD (FelCWD) was transmitted to all IC-inoculated cats with a decreased incubation period of 23 to 27 mo. FelCWD was detected in the brains of all the symptomatic cats by western blotting and immunohistochemistry and abnormalities were seen in magnetic resonance imaging, including multifocal T2 fluid attenuated inversion recovery (FLAIR) signal hyper-intensities, ventricular size increases, prominent sulci, and white matter tract cavitation. Currently, 3 of 4 IP/SQ and 2 of 4 PO inoculared cats have developed abnormal behavior patterns consistent with the early stage of feline CWD. *** These results demonstrate that CWD can be transmitted and adapted to the domestic cat, thus raising the issue of potential cervid-to- feline transmission in nature.


www.prion2013.ca/tiny_uploads/forms/Scientific-Program.pdf


www.landesbioscience.com


Monday, August 8, 2011

Susceptibility of Domestic Cats to CWD Infection

felinespongiformencephalopathyfse.blogspot.com/2011/08/susceptibility-of-domestic-cats-to-cwd.html


Envt.07:

Pathological Prion Protein (PrPTSE) in Skeletal Muscles of Farmed and Free Ranging White-Tailed Deer Infected with Chronic Wasting Disease

Martin L. Daus,1,† Johanna Breyer,2 Katjs Wagenfuehr,1 Wiebke Wemheuer,2 Achim Thomzig,1 Walter Schulz-Schaeffer2 and Michael Beekes1 1Robert Koch Institut; P24 TSE; Berlin, Germany; 2Department of Neuropathology, Prion and Dementia Research Unit, University Medical Center Göttingen; Göttingen, Germany †Presenting author; Email: [e-mail]

Chronic wasting disease (CWD) is a contagious, rapidly spreading transmissible spongiform encephalopathy (TSE) occurring in cervids in North America. Despite efficient horizontal transmission of CWD among cervids natural transmission of the disease to other species has not yet been observed. Here, we report a direct biochemical demonstration of pathological prion protein PrPTSE and of PrPTSE-associated seeding activity in skeletal muscles of CWD-infected cervids. The presence of PrPTSE was detected by Western- and postfixed frozen tissue blotting, while the seeding activity of PrPTSE was revealed by protein misfolding cyclic amplification (PMCA). The concentration of PrPTSE in skeletal muscles of CWD-infected WTD was estimated to be approximately 2000- to 10000-fold lower than in brain tissue. Tissue-blot-analyses revealed that PrPTSE was located in muscle- associated nerve fascicles but not, in detectable amounts, in myocytes. *** The presence and seeding activity of PrPTSE in skeletal muscle from CWD-infected cervids suggests prevention of such tissue in the human diet as a precautionary measure for food safety, pending on further clarification of whether CWD may be transmissible to humans.

www.landesbioscience.com/journals/prion/Prion5-Supp-PrionEnvironment.pdf?nocache=1333529975


Sunday, July 21, 2013

*** As Chronic Wasting Disease CWD rises in deer herd, what about risk for humans?

chronic-wasting-disease.blogspot.com/2013/07/as-chronic-wasting-disease-cwd-rises-in.html


Wednesday, September 04, 2013

***cwd - cervid captive livestock escapes, loose and on the run in the wild...

chronic-wasting-disease.blogspot.com/2013/09/cwd-cervid-captive-livestock-escapes.html


Saturday, October 19, 2013

***ACA Council Meets to Endorse Several Proposed USAHA Resolutions (CWD TSE PRION DISEASE)

chronic-wasting-disease.blogspot.com/2013/10/aca-council-meets-to-endorse-several.html


Thursday, September 19, 2013

Illinois Chronic Wasting Disease: 2012-2013 Surveillance and Management Report

chronic-wasting-disease.blogspot.com/2013/09/illinois-chronic-wasting-disease-2012.html


Monday, December 02, 2013

WISCONSIN CHRONIC WASTING DISEASE CWD DISCOVERED MARATHON COUNTY HUNTING PRESERVE

chronic-wasting-disease.blogspot.com/2013/12/wisconsin-chronic-wasting-disease-cwd.html


Tuesday, December 17, 2013

Wisconsin Second CWD positive deer found in Grant County

chronic-wasting-disease.blogspot.com/2013/12/wisconsin-second-cwd-positive-deer.html


Sunday, December 08, 2013

IOWA DNR to Continue Surveillance for Chronic Wasting Disease CWD TSE PRION DISEASE

chronic-wasting-disease.blogspot.com/2013/12/iowa-dnr-to-continue-surveillance-for.html


Wednesday, August 21, 2013

IOWA DNR EMERGENCY CONSENT ORDER IN THE MATTER OF TOM & LINDA BRAKKE D/B/A PINE RIDGE HUNTING LODGE UPDATE AUGUST 21, 2013

chronic-wasting-disease.blogspot.com/2013/08/iowa-dnr-emergency-consent-order-in.html


Wednesday, October 23, 2013

Steve Lightfoot: West Texas Mule Deer rules CWD Management Plan mandatory check stations for harvested mule deer taken inside the CWD Containment Zone

chronic-wasting-disease.blogspot.com/2013/10/steve-lightfoot-west-texas-mule-deer.html



Thursday, October 03, 2013



TAHC ADOPTS CWD RULE THAT the amendments REMOVE the requirement for a specific fence height for captives



Texas Animal Health Commission (TAHC)



ANNOUNCEMENT



October 3, 2013



chronic-wasting-disease.blogspot.com/2013/10/tahc-adopts-cwd-rule-that-amendments.html



From: [e-mail]
Sent: Tuesday, November 26, 2013 8:16 AM
To: [e-mail]
Subject: RE: Wasting disease is threat to the entire UK deer population





Description: Description: Description: Description: Description: Description: Description: Description: Description: Description: Corp ID for Signatures



Dear Mr Singeltary,



Thank you for your email, which was received by Public Information at the Scottish Parliament. We provide impartial information about the Scottish Parliament, its membership, business and procedures.



The Scottish Parliament’s Rural Affairs, Climate Change and Environment Committee has been looking into deer management, as you can see from the following press release, ***and your email has been forwarded to the committee for information:



www.scottish.parliament.uk/parliamentarybusiness/CurrentCommittees/29878.aspx



You may be interested in following the committee’s work. Papers for committee meetings and transcripts of past meetings (called Official Reports) appear on the following webpage:



www.scottish.parliament.uk/parliamentarybusiness/CurrentCommittees/29878.aspx



I hope this information is of use to you. Please contact us again if you have any questions about the Scottish Parliament.



Yours sincerely,

George Clark
Public Information and Publications
The Scottish Parliament


snip...see full text ;



Friday, November 22, 2013


*** Wasting disease is threat to the entire UK deer population Chronic Wasting Disease CWD TSE prion

chronic-wasting-disease.blogspot.com/2013/11/wasting-disease-is-threat-to-entire-uk.html


Friday, November 29, 2013

Identification of Misfolded Proteins in Body Fluids for the Diagnosis of Prion Diseases

International Journal of Cell Biology

transmissiblespongiformencephalopathy.blogspot.com/2013/11/identification-of-misfolded-proteins-in.html


Sunday, November 3, 2013

Environmental Impact Statements; Availability, etc.: Animal Carcass Management [Docket No. APHIS-2013-0044]

transmissiblespongiformencephalopathy.blogspot.com/2013/11/environmental-impact-statements.html



Sunday, December 15, 2013

*** FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEED VIOLATIONS OFFICIAL ACTION INDICATED OAI UPDATE DECEMBER 2013 UPDATE

madcowusda.blogspot.com/2013/12/fda-part-589-substances-prohibited-from.html




Merry Christmas !



kind regards, terry

[flounder9]

*** PRICE OF CWD TSE PRION POKER GOES UP 2014 ***


Transmissible Spongiform Encephalopathy TSE PRION update January 2, 2014



*** chronic wasting disease, there was no absolute barrier to conversion of the human prion protein.


*** Furthermore, the form of human PrPres produced in this in vitro assay when seeded with CWD, resembles that found in the most common human prion disease, namely sCJD of the MM1 subtype.


wwwnc.cdc.gov/eid/article/20/1/13-0858_article.htm


chronic-wasting-disease.blogspot.com/2014/01/molecular-barriers-to-zoonotic.html


*** 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.


cdmrp.army.mil/prevfunded/nprp/NPRP_Summit_Final_Report.pdf




FINALLY, 12 years later, the OIE becomes concerned with CWD to humans, not that I did not try and warn them 12 years ago. ...kind regards, terry


Friday, January 17, 2014

Annual report of the Scientific Network on BSE-TSE EFSA, Question No EFSA-Q-2013-01004, approved on 11 December 2013


*** Further, it was addressed that recently discussions have being held at OIE level on Chronic Wasting Disease of cervids.


2002 Singeltary vs O.I.E. on CWD to human risk factor ;


Subject: Re: CWD AMERICA

Date: Fri, 12 Jul 2002 19:10:18 +0200

From: "INFORMATION DEPT"

Organization: O.I.E

To: "Terry S. Singeltary Sr."

References: <[e-mail]> <012901c229b2$ad43bb90$7f00000a@HPKB> [e-mail]

I agree with you Dr Terry. The OIE, namely the International Animal Health Code Commission is working on making proposals to Member Countries to change the OIE lists so to avoid some the problems mentioned in you e-mail. This will take at least two years before adoption by the International Committee. For BSE, countries asked the OIE to post information on BSE on the OIE web site.

Personally, I am interested in Chronic Wasting Disease and I follow what is distributed through ProMed. Delegates of OIE Member Countries can propose diseases to be added to the list.

Kind regards.

Karim Ben Jebara


transmissiblespongiformencephalopathy.blogspot.com/2012/07/oie-bse-cwd-scrapie-tse-prion-disease.html


----- Original Message -----

From: "Terry S. Singeltary Sr."

To: "INFORMATION DEPT"

Sent: Friday, July 12, 2002 8:43 PM

Subject: Re: CWD AMERICA


>>> *** Further, it was addressed that recently discussions have being held at OIE level on Chronic Wasting Disease of cervids. <<<


> hello Dr. Jebara,
>
> many thanks for your swift and kind reply.
>
> if i am not mistaken, it was the same email address.
> it was 3 or 4 weeks ago i wrote, as it is, i don't
> save 'sent' emails anymore, unless very important.
>
> my main concern (besides the fact that a potential TSE
> has been in the USA cattle for some time, but the APHIS
> do not test to find), is that the CWD could very well be
> transmitting to humans, and i just did not see to much
> posted about it on OIE site.
>
> > Coming back to your question, Chronic Wasting Disease is not an OIE
>
> > listed disease. Please see OIE disease lists at
>
> www.oie.int/eng/maladies/en_classification.htm#ListeA).
>
> why is this TSE (CWD) not listed and followed as with BSE ?
>
> Article 1.1.3.2.
> 1. Countries shall make available to other countries, through the
> OIE, whatever information is necessary to minimise the spread of
> important animal diseases and to assist in achieving better worldwide
> control of these diseases.
>
> www.oie.int/eng/normes/MCode/A_00005.htm
>
> The USA CWD is an important animal disease.
>
> why is it not followed?
>
> > The decision to add or delete a disease from the OIE lists, come
>
> > through proposals made by Member Countries and it has to be adopted by
>
> > the International Committee.
>
> i _urgently_ suggest a proposal to the OIE to follow this disease very
> closely, and to propose _more_ testing in the USA for TSEs in the USA
> cattle...
>
> kindest regards,
> terry
>
> INFORMATION DEPT wrote:
>
> > Dear Sir,
> >
> > This is the first time that I receive your e-mail. To whom have you written
> > in the OIE or to which address?
> >
> > Coming back to your question, Chronic Wasting Disease is not an OIE listed
> > disease. Please see OIE disease lists at
> > www.oie.int/eng/maladies/en_classification.htm#ListeA).
> >
> > Countries should report to the OIE any disease even is not listed in the
> > OIE's lists in some conditions (example: an exceptional epidemiological
> > event). Please read Chapter 1.1.3 of the International animal health code to
> > have more information on disease notification and epidemiological
> > information agreed by OIE Member Countries at :
> > www.oie.int/eng/normes/MCode/A_00005.htm
> >
> > The decision to add or delete a disease from the OIE lists, come through
> > proposals made by Member Countries and it has to be adopted by the
> > International Committee.
> >
> > Hope that I answered to your question.
> >
> > Best regards.
> >
> > Dr Karim Ben Jebara
> > Head
> > Animal Health Information Department
> > OIE
> >
> >
> >
> > ----- Original Message -----
> > From: "Terry S. Singeltary Sr."
> > To:
> > Sent: Friday, July 12, 2002 6:18 PM
> > Subject: CWD AMERICA
> >
> >
> >
> >>I WROTE TO OIE RECENTLY ASKING 'WHY OIE DOES NOT FOLLOW CWD IN
> >>AMERICA' ? with no reply ? i am still seeking an answer ?
> >>
> >>many thanks,
> >>and kind regards,
> >>terry
=====================


transmissiblespongiformencephalopathy.blogspot.com/2012/07/oie-bse-cwd-scrapie-tse-prion-disease.html


for anyone interested, please see full report here ;

Friday, January 17, 2014

Annual report of the Scientific Network on BSE-TSE EFSA, Question No EFSA-Q-2013-01004, approved on 11 December 2013

efsaopinionbseanimalprotein.blogspot.com/2014/01/annual-report-of-scientific-network-on.html


Thursday, January 2, 2014

*** CWD TSE Prion in cervids to hTGmice, Heidenhain Variant Creutzfeldt-Jacob Disease MM1 genotype, and iatrogenic CJD ***

transmissiblespongiformencephalopathy.blogspot.com/2014/01/cwd-tse-prion-in-cervids-to-htgmice.html


Wednesday, January 01, 2014

APHIS-2006-0118-0100 Chronic Wasting Disease Herd Certification Program and Interstate Movement of Farmed or Captive Deer, Elk, and Moose

chronic-wasting-disease.blogspot.com/2014/01/aphis-2006-0118-0100-chronic-wasting.html


Wednesday, January 15, 2014

INFECTION PREVENTION AND CONTROL OF CJD, VCJD AND OTHER HUMAN PRION DISEASES IN HEALTHCARE AND COMMUNITY SETTINGS Variably Protease-Sensitive Prionopathy (VPSPr) January 15, 2014

transmissiblespongiformencephalopathy.blogspot.com/2014/01/infection-prevention-and-control-of-cjd.html




kind regards,
terry