PERSON TO PERSON TRANSMISSION OF THE TSE PRION DISEASE, never say never. as the disease mutates, it becomes more virulent in some cases, and cwd is efficiently transmitted from cervid to cervid. there are now multiple strains of CWD in cervids, as with the TSE prion disease in bovine, sheep and goats, and we now have the atypical TSE in these species, that have mutated, and some strains _have_ become more virulent. we now have younger humans dying from the TSE prion disease, with shorter incubation period, and that are much younger. human to human casual transmission of the TSE prion disease...again, never say never. ...TSS
A Kiss of a Prion: New Implications for Oral Transmissibility
Bianca Da Costa Dias and Stefan F. T. Weiss + Author Affiliations
School of Molecular and Cell Biology University of the Witwatersrand, 
Johannesburg South Africa Reprints or correspondence: Prof Stefan F. T. Weiss 
School of Molecular and Cell Biology University of the Witwatersrand Private Bag 
3 2050 Wits Johannesburg South Africa (stefan.weiss@wits.ac.za). There is no 
doubt about it: prions-infectious particles composed mainly if not entirely of 
misfolded protein (scrapie-type prion protein [PrPSc]), which are the causative 
agents of transmissible spongiform encephalopathies (TSE) such as scrapie, 
variant Creutzfeldt-Jakob Disease (vCJD), and bovine spongiform encephalopathy 
(BSE)-are transmissible [1–3]. These agents may be introduced via intracerebral, 
intravenous, intraperitoneal, or intraventricular infection, and recent research 
indicates that oral transmission may also occur. The last mode of transmission 
is of particular interest because it indicates that the consumption of meat and 
other products derived from animals experiencing prion disorders may pose a real 
risk to humans. Recent reports suggest that, in addition to meat, bodily fluids 
such as blood, saliva, feces, and milk may well be risk factors for possible 
transmission of TSEs to humans. Successful oral transmission among different 
animal species (interspecies) has been demonstrated. However, species 
specificity, the “ species barrier,” and the mode of transmission must be taken 
into account and may explain why cattle, sheep, goats, mink, and mice are 
successfully orally infected with bovine scrapie-type prion protein (bovPrPSc), 
whereas the ingestion of bovPrPSc by pigs, poultry, and cervids such as elk and 
deer fails to cause disease [1]. Humans are also thought to be susceptible to 
oral infection by bovPrPSc by means of contaminated bovine products (eg, meat 
pies), and this is believed to be the manner in which the zoonotic disease vCJD 
originated [4]. 
But where do prions hide in the body? They replicate primarily in the 
central nervous system, particularly in the brain and the lymphoreticular system 
[4], as well as in other tissues such as muscle [5]. Furthermore, the presence 
of these infectious agents in bodily excretions and secretions is a major cause 
for concern, because it enhances the risks of transmissibility. Prions have been 
identified in feces of asymptomatic deer [6] and in the blood, saliva, and urine 
of deer with chronic wasting disease [7, 8]. PrPSc has also been detected in the 
salivary glands of scrapie-affected sheep [9]. 
The report by Maddison et al [10] in this issue of theJournal describes for 
the first time, to our knowledge, the secretion of prions into the oral cavity 
of sheep. The authors used silicon dioxide (SiO2) to concentrate prions, in 
conjunction with serial protein misfolding cyclic amplification, or sPMCA, a 
method to amplify and detect the presence of very low concentrations of PrPSc. 
Serial protein misfolding cyclic amplification has numerous applications, such 
as the sensitive detection of pathological prions [11], later application for in 
vitro generation of prions [12], and detection of prions in body fluids such as 
blood from scrapie-infected hamsters [13]; the last example succeeded even in 
the presymptomatic phase [14]. Maddison et al [10] used this technique to 
demonstrate that prions are present in buccal swab samples obtained from sheep 
with preclinical scrapie infections. 
However, one must pose the following question: how do ingested infectious 
PrPSc prions reach the mucus and saliva? After oral ingestion, prions are 
thought to be taken up first by Peyer patches before they disseminate through 
gut-associated lymphoid tissues, the lymphoreticular system, the vagus nerve, 
and the enteric nervous system, after which they enter the central nervous 
system [15]. Internalization of prions in the intestine is thought to be 
performed by M-(microfold) cells [16] and by enterocytes, which internalize 
bovPrPSc dependent on the prion receptor LRP/LR [17]. 
Maddison et al [10] suggest, according to their data, that prions are able 
to spread from the small intestine to the oral salivary glands and epithelia 
within a period of 9 months. This route explains the occurrence of prions in 
saliva and the shedding of prions into the oral cavity. 
The transmissibility of scrapie among sheep (intraspecies) is well 
recognized. It must be emphasized that horizontal transfer (from one individual 
to another) of scrapie is the main route of infection, because vertical 
transmission of disease from mother to offspring via milk or placental tissue 
occurs infrequently. Thus, in view of the report by Maddison et al, the oral 
transmissibility of prions among sheep may serve as a major route for horizontal 
scrapie transfer. This occurrence is plausible because sheep often lick each 
other. Maddison et al [10] indicate that, because of the similarities in prion 
tissue distribution, their implications for the oral transmission of ovine 
scrapie might be true for other prion diseases, such as cervid chronic wasting 
disease and human vCJD. If this is true for humans, a kiss of a prion may 
sometimes have lethal consequences. 
 Next Section Acknowledgments We thank the Deutsche Forschungsgemeinschaft 
(DFG) grant WE 2664/2–1, Germany and the National Research Foundation (NRF), 
South Africa, for financial support. We thank Professor Juergen Richt, Kansas 
State University, United States, for a critical reading of this paper. 
Previous SectionNext Section Footnotes Potential conflicts of interest: 
none reported.
Financial support: Deutsche Forschungsgemeinschaft, Germany (grant WE 
2664/2–1) and the National Research Foundation, South Africa. 
 we are (in my opinion), exposed to the TSE prion in so many different ways 
in every day life, the potential for exposure and then becoming infected via 
taking care of a loved one with TSE prion disease, in my opinion risk factor 
there from is minimal, if proper precautions are taken, and even if they were 
not, the chance of becoming infected from a kiss, or casual contact is low, but 
I do not think it is zero, actually, far from it. I put this all together for a 
documentation of the known facts to date, of the potential casual human to human 
transmission. I did not put it together to scare anyone. with aerosol 
transmission of the TSE prion a reality now, infectivity in urine and feces and 
transmission there from being reality now with the TSE prion disease, I don’t 
see how anyone can rule _out_ the potential for transmission of the TSE prion 
via a kiss (a vehicle for transmission of the TSE prion via saliva), or even for 
a cut or open wound (all a cut is and transmission there from, is an crude 
inoculation of sorts, and inoculation has been proven to be an efficient mode of 
transmission for the TSE prion disease), even the eye, from either one of the 
body fluids now that how proven to be infections. I can’t see why we have such 
safety protocols for laboratory workers working with the TSE prion disease, but 
yet the same officials will say it’s o.k. for the public, friends, and or family 
members to do just the opposite with their loved ones when succumbing to the CJD 
TSE prion disease. don’t get me wrong, I did it too, and would probably do it 
again as far as kissing my mom. but science is science, and the transmission 
studies speak for themselves with the bodily fluids. simply put, which is all I 
was saying, we can’t say never, and or that none of these cases to date, have 
not been, and or will not be, a potential vehicle for transmission. I believe, 
and this is my opinion, that more concern for casual transmission with body 
fluids and materials there from, should be put forth to families with their 
loved ones, and I think that the safety protocols there from should be revised, 
to match that of the laboratory settings. again, this is my opinion. your 
opinion, and or others here, may read the same science and feel different out 
the findings. ...take care, kind regards, terry 
 SUBSTANCE DATA SHEET
HUMAN PRION AGENTS
FOR USE IN RESEARCH LABORATORIES
SECTION I - INFECTIOUS AGENT
Name: Creutzfeldt-Jakob agent, Kuru agent
Synonym or Cross Reference:: Subacute spongiform encephalopathy, 
Creutzfeldt-Jakob disease (CJD), Kuru, Transmissible Spongiform Encephalopathy 
(TSE).
Characteristics: Filterable, self-replicating agent, slow infectious 
pathogen, prion protein (PrP)
SECTION II - RECOMMENDED PRECAUTIONS
Containment Requirements: Biosafety level 3 facilities, practices and 
containment equipment for activities involving these agents; also listed under 
biosafety level 2 with special precautions; level of containment will depend on 
the nature of the manipulations and the amount of sera, bio/necropsy materials 
handled
Protective Clothing: Gown and gloves when handling potentially infectious 
materials; eye protection may also be indicated
Other Precautions: Extreme care must be taken to avoid accidental 
autoinoculation or other parenteral inoculations of infectious tissues and 
fluids
SECTION III - HANDLING INFORMATION
Spills: Allow any potential aerosols to settle; wearing protective 
clothing, gently cover spill with paper towel and apply 1N sodium hydroxide, 
starting at perimeter and working towards the center; allow sufficient contact 
time (1 hour) before clean up
Disposal: Decontaminate before disposal; steam sterilization (132·C for 1 
hour), disinfection with 1N sodium hydroxide for 1 hour, incineration
Storage: In sealed containers that are appropriately labeled 
The main precaution to be taken by laboratorians working with 
prion-infected or contaminated material is to avoid accidental puncture of the 
skin.3 Persons handling contaminated specimens should wear cut-resistant gloves 
if possible. If accidental contamination of unbroken skin occurs, the area 
should be washed with detergent and abundant quantities of warm water (avoid 
scrubbing); brief exposure (1 minute to 1N NaOH or a 1:10 dilution of bleach) 
can be considered for maximum safety.6 Additional guidance related to 
occupational injury are provided in the WHO infection control guidelines.6 
Unfixed samples of brain, spinal cord, and other tissues containing human prions 
should be processed with extreme care in a BSL-2 facility utilizing BSL-3 
practices. 
Bovine Spongiform Encephalopathy Although the eventual total number of 
variant CJD cases resulting from BSE transmission to humans is unknown, a review 
of the epidemiological data from the United Kingdom indicates that BSE 
transmission to humans is not efficient.9 The most prudent approach is to study 
BSE prions at a minimum in a BSL-2 facility utilizing BSL-3 practices. When 
performing necropsies on large animals where there is an opportunity that the 
worker may be accidentally splashed or have contact with high-risk materials 
(e.g., spinal column, brain) personnel should wear full body coverage personal 
protective equipment (e.g., gloves, rear closing gown and face shield). 
Disposable plasticware, which can be discarded as a dry regulated medical waste, 
is highly recommended. Because the paraformaldehyde vaporization procedure does 
not diminish prion titers, BSCs must be decontaminated with 1N NaOH and rinsed 
with water. HEPA filters should be bagged out and incinerated. Although there is 
no evidence to suggest that aerosol transmission occurs in the natural disease, 
it is prudent to avoid the generation of aerosols or droplets during the 
manipulation of tissues or fluids and during the necropsy of experimental 
animals. It is further strongly recommended that impervious gloves be worn for 
activities that provide the opportunity for skin contact with infectious tissues 
and fluids. 
The main precaution to be taken when working with prion-infected or 
contaminated material is to avoid puncture of the skin. If accidental 
contamination of skin occurs, the area is swabbed with In sodium hydroxide 
(NaOH) for 5 minutes and then washed with copious amounts of water. Unfixed 
samples of brain, spinal cord, and other tissues containing human prions should 
be processed with extreme care at BSL 3.
Prions are characterized by extreme resistance to conventional inactivation 
procedures including irradiation, boiling, dry heat, and chemicals (formalin, 
betapropiolactone, alcohols). Sterilization of rodent brain extracts with high 
titers of prions requires autoclaving at 132C for 4.5 hours. Denaturing organic 
solvents such as phenol or chaotropic reagents such as guanidine isothiocyanate 
or alkali such as NaOH can also be used for sterilization. Disposable 
plasticware, which can be discarded as a dry waste, is highly recommended.
Although there is no evidence to suggest that aerosol transmission occurs 
in the natural disease, it is prudent to avoid the generation of aerosols or 
droplets during the manipulation of tissues or fluids and during the necropsy of 
experimental animals. Formaldehyde-fixed and paraffin-embedded tissues, 
especially of the brain, remain infectious. Some investigators recommend that 
formalin-fixed tissues from suspected cases of prion disease be immersed for 30 
min in 96% formic acid or phenol before histopathologic processing, but such 
treatment may severely distort the microscopic neuropathology. 
another interesting aspect of the TSE prion disease is KURU ; 
Figure 25. All cooking. including that of human flesh from diseased 
kinsmen. was done in pits with steam made by pouring water over the hot stones, 
or cooked in bamboo cylinders in the hot ashes. Children participated in both 
the butchery and the handling of cooked meat, rubbing their soiled hands in 
their armpits or hair, and elsewhere on their bodies. They rarely or never 
washed. Infection with the kuru virus was most probably through the cuts and 
abrasions of the skin. or from nose-picking, rye (eye...tss) rubbing, or mucosal 
injury.
These detailed descriptions will be published elsewhere but have reaffirmed 
the oral histories of endocannibalism in the Fore recorded previously12,22–24 
and that this practice ceased abruptly at the time of Australian administrative 
control over the kuru areas. Although isolated events might have occurred for a 
few years after this prohibition, we are confident that new exposures of 
individuals to kuru at mortuary feasts would not have occurred after 1960. Not 
only have no cases of kuru been recorded in people born after 1959 (and only 
nine were recorded in those born after 1956); but also all the 11 last recorded 
cases of kuru that we report here were born before 1950. If any source of 
infection remained, whether from surreptitious cannibalism, possible ground 
contam-ination with human prions at sites where food was prepared, or other 
lateral routes, we would expect individuals born after this period to have 
kuru—especially since children are thought to have had shorter incubation 
periods than adults. However, no such cases have been observed. Additionally, 
although a fraction of hamster-adapted scrapie prions have been shown to survive 
in soil for at least 3 years,25 the mortuary feast practices (during which the 
entire body would be consumed) were undertaken so that any substantial 
contamination of soil would not have occurred, and traditional bamboo knives and 
leaf plates were burned after the feast. Furthermore, no clusters of kuru cases, 
as seen earlier in the epidemic,26 have been recorded for many years.... 
Kuru: The Science and the Sorcery 
Special Jury Prize Winner, Pacific International Documentary Film Festival 
2011.
This is the true story of one of the most incredible and challenging 
medical detective stories of the 20th Century; a history of human tragedy, 
adventure and discovery. It is the story of the Fore, a Papuan community 
immersed in cannibalistic mortuary practices and sorcery in one of the most 
remote regions on the planet, and the tragic disease that threatened to wipe out 
their entire population. 
In 1961, a young Australian medical researcher, Michael Alpers, puts up his 
hand to work on a new and strange disease in the Eastern Highlands of Papua New 
Guinea. There, he teams up with an American outer, Dr Carleton Gajdusek, who has 
been in the local Fore region since 1957. For Michael it is the beginning of a 
lifelong obsession.
Together, they are amidst a major epidemic. It is killing over 200 people a 
year with devastating effects. It mainly targets women and children. The local 
people, the Fore, call the disease kuru, their word for shivering. They believe 
it is caused by sorcery.
Michael and Carleton are baffled by the disease. There are no scientific 
disciplines to guide them as they attempt to unravel its mysteries. By pure 
chance, a link is made to a strange transmissible animal disease in sheep, 
Scrapie. The two kuru researchers embark on a 10-year experiment to see if the 
fatal degenerative brain disease in humans could be transmissible like 
Scrapie.
The decision is made to perform an autopsy on a kuru victim and inoculate 
the kuru material into a chimpanzee. Kigea, ayoung girl in the village is 
identified as being in the early stages of kuru. Kigea’s family, gives Michael 
permission to perform an autopsy upon her death.
A brain sample taken from Kigea after her death is flown to the USA and 
injected into a chimpanzee called Daisy. While Michael follows the progress of 
the transmission experiment, he starts to collate all the recorded data on kuru 
and begins to suspect cannibalism as the cause of the spreadof the 
disease.
Within two years, he diagnoses Daisy with kuru. This is a defining moment. 
It confirms kuru is transmissible and can cross the species barrier. The 
revelation, together with epidemiological data collated with anthropologist 
Shirley Lindenbaum, links the Fore’s mortuary feasts (consumption of dead 
relatives) to the transmission of kuru. Cannibalism is the cause, and its origin 
is linked to a rare disease called Creutzfeldt Jakob Disease(CJD), but the story 
of kuru is far from over.
The infecting agent is the first new pathogen – prions – to be discovered 
in over 100 years. Research results in two Nobel prizes: it’s discoveries 
turning scientific understanding upside down, causing rifts in the beliefs ofthe 
science community.
Then Mad Cow Disease (Bovine Spongiform Encephalopathy or BSE) reared its 
head in the mid 1980s, and 10 years later the human variant CJD. All eyes turned 
to kuru, the only model of a prion epidemic in human populations. Many unknowns 
still surround prion diseases: there is no cure for kuru, or any of the prion 
diseases. The effects are devastating and unprecedented incubation periods can 
extend beyond 50 years.
Michael is the key and heart to this story, providing unique access to the 
Fore people, and the world’s other leading authorities on the matter; including 
Americans Prof. DC Gajdusek (Nobel Prize 1976), Prof. Stan Prusiner (Nobel Prize 
1997), Prof Shirley Lindenbaum (Anthropologist) and British Prof. John Collinge 
(Director, MRC Prion Unit, UK). 
Kuru: The Science and the Sorcery combines history, science and 
anthropology to tell a unique and ongoing ‘history of science’ documentary 
spanning five decades. It intertwines the thinking of great minds, locally and 
internationally, to reveal how this rare disease in the remote highlands of PNG 
exploded to international attention and how Prion research has now revealed we 
are all descendants of a remote past of cannibal practices. 
Kuru: The Science and the Sorcery Australian scientist Michael Alpers 
dedicated over 50 years to researching Kuru, an obscure and incurable brain 
disease unique to the Fore people of New Guinea. Kuru was once thought to be a 
psychosomatic illness, an infection, a genetic disorder, even a sorcerer's 
curse, but Alpers' findings pointed to cannibalism as the culprit. Yet a recent 
discovery has proven to be even more disturbing: the malady is linked to mad cow 
disease and its human equivalent, variant CJD. With a decades-long incubation 
period, could a larger outbreak be on its way?
human flesh taste very sweet
KURU EPIDEMIOLOGICAL PATROLS 
Michael Alpers
First Reports
People of the Kuru region part 1
boy playing with animal bladder, blowing it up like a balloon. ...
People of the Kuru region part 2 
Monday, November 19, 2012 
Prion in Saliva of Bovine Spongiform Encephalopathy–Infected Cattle 
 Aerosols
Prion transmission is usually not considered to be airborne like influenza 
or chicken pox. But we and others recently have found that prions can also be 
efficiently transmitted to mice through aerosols [5], [6]. Although 
aerosol-transmitted prions have never been found under natural conditions, this 
finding highlights the necessity of revising the current prion-related biosafety 
guidelines and health standards in diagnostic and scientific laboratories being 
potentially confronted with prion-infected materials.
Thursday, December 29, 2011 
Aerosols An underestimated vehicle for transmission of prion diseases? 
PRION 
please see more on Aerosols and TSE prion disease here ;
Monday, November 26, 2012 
Aerosol Transmission of Chronic Wasting Disease in White-tailed Deer 
Simultaneous Onset of Alzheimer's Disease in a Husband and Wife in Their 
Mid Fifties: What do We Really Know? 
 Jonathan Heath1, Lindsay Goicochea2, Mark Smith3, Rudy Castellani4. 
1Department of Pathology, University of Maryland; 2University; 3Case Western 
Reserve University; 4University of Maryland, Baltimore, Maryland 
 Whereas the genetic factors influencing the development and expression of 
Alzheimer's disease are well characterized, environmental factors are currently 
thought to play a marginal role. Such factors as prior closed head injury, 
post-menopausal estrogen deficiency, aluminum exposure, smoking, diabetes, 
atherosclerotic cardiovascular disease, and diet, among others, confer only a 
modest increased risk if any, and are only tangentially considered in the major 
pathogenic cascades that are presently hypothesized. We present the simultaneous 
onset of Alzheimer's disease in a husband and wife, with both subjects 
experiencing cognitive dysfunction within the same month. Both subjects were in 
their mid-fifties at the time of presentation, both subjects showed 
progressively neurological decline with prominent memory loss, both subjects 
experienced myoclonus late in their disease course prompting referral to the 
National Prion Disease Pathology Surveillance Center, and both subjects expired 
12 years after onset, within two months of each other. Review of the family 
pedigree revealed no family history of dementia or other neurologic illnesses in 
multiple first degree relatives. The only historical finding of note was that 
both subjects had moved out of their home briefly while it was being remodeled, 
and both became symptomatic shortly after moving back in. At autopsy, the 
subjects had classic advanced Alzheimer's disease, with Braak stage VI pathology 
that was otherwise identiical in quantity and distribution of amyloid-beta, 
cerebral amyloid angiopathy, and neurofibrillary degeneration. While no specific 
toxin or other environmental cause was discerned, these two cases raise the 
issue of epigenetic factors in Alzheimer's disease that may be more robust than 
current literature indicates. 
NEUROLOGY 1998;50:684-688 © 1998 American Academy of Neurology
Creutzfeldt-Jakob disease in a husband and wife 
 P. Brown, MD, L. Cervenáková, MD, L. McShane, PhD, L. G. Goldfarb, MD, K. 
Bishop, BS, F. Bastian, MD, J. Kirkpatrick, MD, P. Piccardo, MD, B. Ghetti, MD 
and D. C. Gajdusek, MD From the Laboratory of CNS Studies (Drs. Brown, 
Cervenáková, Goldfarb, and Gajdusek), NINDS, and Biometric Research Branch (Dr. 
McShane), NCI, National Institutes of Health, Bethesda, MD; the Department of 
Obstetrics (K. Bishop), Gynecology and Reproductive Sciences, University of 
Texas Houston Health Science Center, Houston, TX; the Department of Pathology 
(Dr. Bastian), University of South Alabama Medical Center, Mobile, AL; the 
Department of Pathology (Dr. Kirkpatrick), The Methodist Hospital, Houston, TX; 
and the Department of Pathology (Drs. Piccardo and Ghetti), Indiana University 
School of Medicine, Indianapolis, IN.
Address correspondence and reprint requests to Dr. Paul Brown, Building 36, 
Room 5B21, National Institutes of Health, Bethesda, MD 20892. 
A 53-year-old man died of sporadic Creutzfeldt-Jakob disease (CJD) after a 
1.5-year clinical course. Four and a half years later, his then 55-year-old 
widow died from CJD after a 1-month illness. Both patients had typical clinical 
and neuropathologic features of the disease, and pathognomonic 
proteinase-resistant amyloid protein ("prion" protein, or PrP) was present in 
both brains. Neither patient had a family history of neurologic disease, and 
molecular genetic analysis of their PrP genes was normal. No medical, surgical, 
or dietary antecedent of CJD was identified; therefore, we are left with the 
unanswerable alternatives of human-to-human transmission or the chance 
occurrence of sporadic CJD in a husband and wife. 
 --------------------------------------------------------------------------------
Received May 5, 1997. Accepted in final form September 10, 1997. 
SEE CJD IN MAN AND HIS CAT ; 
 [Image] Research letters Volume 352, Number 9134 [Image] 3 October 1998 
[Previous] [Next]
[Image][Image]
Simultaneous occurrence of spongiform encephalopathy in a man and his cat 
in Italy
[Image]
Gianluigi Zanusso, Ettore Nardelli, Anna Rosati, GianMaria Fabrizi, Sergio 
Ferrari, Antonella Carteri, Franco De Simone, Nicola Rizzuto, Salvatore 
Monaco
Transmissible spongiform encephalopathies (TSE) encompass inherited, 
acquired, and sporadic mammalian neurological disorders, and are characterised 
by the conversion of the cellular prion protein (PrP) in an insoluble and 
protease-resistant isoform (PrPres). In human TSE, four types of PrPres have 
been identified according to size and glycoform ratios, which may represent 
different prion strains. Type-1 and type-2 PrPres are associated with sporadic 
Creutzfeldt-Jakob disease (CJD), type 3 with iatrogenic CJD, and type 4 with 
variant CJD.1,2 There is evidence that variant CJD is caused by the bovine 
spongiform encephalopathy (BSE)-prion strain.2-4 The BSE strain has been 
identified in three cats with feline spongiform encephalopathy (FSE), a prion 
disease which appeared in 1990 in the UK.5 We report the simultaneous occurrence 
of sporadic CJD in a man and a new variety of FSE in his cat.
A 60-year-old man, with no unusual dietary habits, was admitted in 
November, 1993, because of dysarthria, cerebellar ataxic gait, visual agnosia, 
and myoclonus. An electroencephalogram (EEG) showed diffuse theta-delta 
activity. A brain magnetic resonance imaging scan was unremarkable. 10 days 
later, he was speechless and able to follow only simple commands. Repeat EEGs 
showed periodic triphasic complexes. 2 weeks after admission, he was mute, 
akinetic, and unable to swallow. He died in early January, 1994.
His 7-year-old, neutered, female shorthaired cat presented in November, 
1993, with episodes of frenzy, twitching of its body, and hyperaesthesia. The 
cat was usually fed on canned food and slept on its owner's bed. No bites from 
the cat were recalled. In the next few days, the cat became ataxic, with 
hindquarter locomotor dysfunction; the ataxia got worse and there was diffuse 
myoclonus. The cat was killed in mid-January, 1994.
No pathogenic mutations in the patient's PrP gene were found. The patient 
and the cat were methionine homozygous at codon 129. Histology of the patient's 
brain showed neocortical and cerebellar neuronal loss, astrocytosis, and 
spongiosis (figure A). PrP immunoreactivity showed a punctate pattern and 
paralleled spongiform changes (figure B). The cat's brain showed mild and focal 
spongiosis in deeper cortical layers of all four lobes (figure C), vacuolated 
cortical neurons (figure D), and mild astrogliosis. The cerebellar cortex and 
the dentate nucleus were gliosed. Immunoreactive PrP showed a punctate pattern 
in neocortex, allocortex, and caudate nucleus (figure E). Western blot analysis 
of control and affected human and cat brain homogenates showed 3 PrP bands of 
27-35 kDa. After digestion with proteinase K and deglycosylation, only samples 
from the affected patient and cat showed type-1 PrPres, with PrP glycoform 
ratios comparable to those observed in sporadic CJD1 (details available from 
author).
[Image]
Microscopic sections of patient and cat brains
A: Occipital cortex of the patient showing moderate spongiform degeneration 
and neuronal loss (haematoxylin and eosin) and B: punctate perineuronal pattern 
of PrP immunoreactivity; peroxidase immunohistochemistry with monoclonal 
antibody 3F4. C: cat parietal cortex showing mild spongiform degeneration 
(haematoxylin and eosin).D: vacuolated neurons (arrow, haematoxylin and eosin), 
E: peroxidase immunohistochemistry with antibody 3F4 shows punctate perineuronal 
deposition of PrP in temporal cortex.
This study shows a spatio-temporal association between human and feline 
prion diseases. The clinical features of the cat were different from previously 
reported cases of FSE which were characterised by gradual onset of behavioural 
changes preceding locomotor dysfunction and ataxia.5 Neuropathological changes 
were also at variance with the diffuse spongiosis and vacuolation of brainstem 
neurons, seen in FSE.5 The synaptic pattern of PrP deposition, similar in the 
cat and in the patient, was atypical for a BSE-related condition. Evidence of a 
new type of FSE was further provided by the detection of a type-1 PrPres, other 
than the BSE-associated type 4.2 Taken together, our data suggest that the same 
agent strain of sporadic CJD was involved in the patient and in his cat.
It is unknown whether these TSE occurred as the result of horizontal 
transmission in either direction, infection from an unknown common source, or 
the chance occurrence of two sporadic forms.
1 Parchi P, Castellani R, Capellari S, et al. Molecular basis of phenotypic 
variablity in sporadic Creutzfeldt-Jakob disease. Ann Neurol 1996; 39: 767-78 
[PubMed].
2 Collinge J, Sidle KCL, Meads J, Ironside J, Hill AF. Molecular analysis 
of prion strain variation and the aetiology of 'new variant' CJD. Nature 1996; 
383: 685-90 [PubMed].
3 Bruce ME, Will RG, Ironside JW, et al. Transmissions to mice indicate 
that 'new variant' CJD is caused by the BSE agent. Nature 1997; 389: 498-501 
[PubMed].
4 Hill AF, Desbruslais M, Joiner S, et al. The same prion strain causes 
vCJD and BSE. Nature 1997; 389: 448-50 [PubMed].
5 Pearson GR, Wyatt JM, Henderson JP, Gruffydd-Jones TJ. Feline spongiform 
encephalopathy: a review. Vet Annual 1993; 33: 1-10.
------------------------------------------------------------------------ 
Sezione di Neurologie Clinica, Dipartimento di Scienze Neurologiche e della 
Visione, Università di Verona, Policlinico Borgo Roma, 37134 Verona, Italy (S 
Monaco; e mail rizzuto@Gorgorna.univr.it); and Istituto Zooprofilattico 
Sperimentale della Lombardia e dell' Emilia, Brescia 
======================================= 
Terry S. Singeltary Sr. wrote: 
######## Bovine Spongiform Encephalopathy #########
Greetings list members,
ODD that some FELINE in Italy seem to have this same or maybe very similar 
phenotype of TSE;
In October 1998 the simultaneous occurrence of spongiform encephalopathy in 
a man and his pet cat was reported. The report from Italy noted that the cat did 
not display the same clinical features as FSE cases previously seen. Indeed, the 
presence of a new type of FSE was suggested. The man was diagnosed as having 
sporadic CJD, and neither case (man nor cat) appeared to be affected by a 
BSE-related condition. 
Case‐to‐case transmission in humans: case reports and series in which 
spread through everyday human contact is suggested There are six reports in 
which this possible mode of transmission is considered. The most recent is that 
of a couple from the USA who had been married for 30 years.47 The husband died 
at age 53. He had no relevant family history, but had had a rotator cuff repair 
one year before disease onset. His wife developed symptoms four and half years 
after her husband's death. She was morbidly obese and had had a previous 
hysterectomy, hernia repair and cholecystectomy. Both occasionally ate brains in 
the form of ‘kizka’, a type of sausage. 
Immunocytochemistry confirmed pathogenic prion protein deposition in brain 
tissue from both husband and wife. Full sequencing of the open reading frame of 
the PRNP failed to demonstrate any pathogenic mutations. Another suspected 
conjugal case has recently been shown not to be CJD. The histopathological 
specimens did not stain for prion protein despite the microscopic appearance of 
spongiform change.48
Sporadic CJD has been described in two co‐workers who shared a school wing 
for 9 months.49 The first was a 48‐year‐old Californian‐born man of Hispanic 
American descent who had had a traumatic leg amputation at age 23, but was 
otherwise well. The second was a 48‐year‐old Chilean‐born male who had a blood 
transfusion 6 months before onset of symptoms, and was known to eat lambs' 
brains. The first patient developed symptoms 5 months after the last contact 
with his colleague and was confirmed to have spCJD 2 months after this. The 
second patient developed symptoms months later and died 9 months after the last 
contact with his colleague. 
An English woman, who died of CJD, histologically confirmed at post mortem, 
was known to have contact with several affected members of a family with 
familial CJD and was related to them by marriage.39 She had known one of the 
family, who later died of CJD and had afternoon tea with her at family 
gatherings, twice a year, for 20 years, as well as visiting in her final 
illness. The woman herself died 12 years later. There is another similar case of 
probable CJD, reported in a Chilean woman who died 13 years after contact with a 
family with familial CJD. No details of contact are given. A third case of death 
from CJD in someone related in marriage to a family with familial CJD has been 
reported in France, in a Tunisian family. No details are given with regards to 
family history or contact.21 What is notable about these last three incidents of 
supposed infection by social contact is that all have occurred in association 
with familial CJD. Although these patients were not known to have been 
genetically related to their spouses, the possibility that they came from the 
same gene pool cannot be dismissed. 
Science 
Coincident Scrapie Infection and Nephritis Lead to Urinary Prion 
Excretion
Seeger, Harald; Heikenwalder, Mathias; Zeller, Nicolas; Kranich, Jan; 
Schwarz, Petra; Gaspert, Ariana; Seifert, Burkhardt; Miele, Gino; Aguzzi, 
Adriano
Date of publication 2005 Language English Notes Includes references Medium 
text Pagination p. 324-326. Journal Title Science ISSN 0036-8075 Volume/Issue 
2005 Oct. 14, v. 310, no. 5746 Abstract Prion infectivity is typically 
restricted to the central nervous and lymphatic systems of infected hosts, but 
chronic inflammation can expand the distribution of prions. We tested whether 
chronic inflammatory kidney disorders would trigger excretion of prion 
infectivity into urine. Urinary proteins from scrapie-infected mice with 
lymphocytic nephritis induced scrapie upon inoculation into noninfected 
indicator mice. Prionuria was found in presymptomatic scrapie-infected and in 
sick mice, whereas neither prionuria nor urinary PrP[superscript Sc] was 
detectable in prion-infected wild-type or PrP[superscript C]-overexpressing 
mice, or in nephritic mice inoculated with noninfectious brain. Thus, urine may 
provide a vector for horizontal prion transmission, and inflammation of 
excretory organs may influence prion spread. 
--------------------------------------------------------------------------------
Source: Information Systems Division, National Agricultural Library (United 
States of America) NAL/USDA 10301 Baltimore Avenue Beltsville, Md. 20705 Email: 
access@nal.usda.gov; URL: http://www.nal.usda.gov/ 
-------------------------------------------------------------------------------- 
AGRIS 2013 - FAO of the United Nations 
Research Article 
Detection of Prion Protein in Urine-Derived Injectable Fertility Products 
by a Targeted Proteomic Approach 
Alain Van Dorsselaer mail, * E-mail: neil.cashman@vch.ca (NRC); 
vandors@unistra.fr (AVD)
Affiliation: Laboratoire de Spectrométrie de Masse Bio-Organique, 
Université de Strasbourg, IPHC, CNRS, UMR7178, Strasbourg, France 
X Christine Carapito, Affiliation: Laboratoire de Spectrométrie de Masse 
Bio-Organique, Université de Strasbourg, IPHC, CNRS, UMR7178, Strasbourg, France 
X François Delalande, Affiliation: Laboratoire de Spectrométrie de Masse 
Bio-Organique, Université de Strasbourg, IPHC, CNRS, UMR7178, Strasbourg, France 
X Christine Schaeffer-Reiss, Affiliation: Laboratoire de Spectrométrie de 
Masse Bio-Organique, Université de Strasbourg, IPHC, CNRS, UMR7178, Strasbourg, 
France 
X Daniele Thierse, Affiliation: Laboratoire de Spectrométrie de Masse 
Bio-Organique, Université de Strasbourg, IPHC, CNRS, UMR7178, Strasbourg, France 
X Hélène Diemer, Affiliation: Laboratoire de Spectrométrie de Masse 
Bio-Organique, Université de Strasbourg, IPHC, CNRS, UMR7178, Strasbourg, France 
X Douglas S. McNair, Affiliation: Cerner Corporation, Kansas City, 
Missouri, United States of America 
X Daniel Krewski, Affiliation: McLaughlin Centre for Population Health Risk 
Assessment, University of Ottawa, Ontario, Canada 
X Neil R. Cashman mail 
snip...
Conclusions/Significance The presence of protease-sensitive prion protein 
in urinary-derived injectable fertility products containing hCG, hMG, and hMG-HP 
suggests that prions may co-purify in these products. Intramuscular injection is 
a relatively efficient route of transmission of human prion disease, and young 
women exposed to prions can be expected to survive an incubation period 
associated with a minimal inoculum. The risks of urine-derived fertility 
products could now outweigh their benefits, particularly considering the 
availability of recombinant products.
Citation: Van Dorsselaer A, Carapito C, Delalande F, Schaeffer-Reiss C, 
Thierse D, et al. (2011) Detection of Prion Protein in Urine-Derived Injectable 
Fertility Products by a Targeted Proteomic Approach. PLoS ONE 6(3): e17815. 
doi:10.1371/journal.pone.0017815
Editor: Jean-Luc Darlix, Institut National de la Santé et de la Recherche 
Médicale, France 
Received: November 12, 2010; Accepted: February 10, 2011; Published: March 
23, 2011 
Discussion Using classical proteomic analyses, prion protein was detected 
for the first time in two u-hCG preparations, and was among 33 different 
non-gonadotropin proteins identified as contaminants of these pharmaceutical 
products. In contrast, r-hCG preparations were negative for prion 
proteins.
In one of the two u-hCG products tested, human prion protein was among the 
ten major contaminants. The fact that prion protein sequences were identified in 
several spots on our 2D electrophoresis gels is likely due to the presence of 
heterogeneous glycosylation and degraded prion protein forms. It is also worth 
noting that in the u-hCG preparation of manufacturer A, plasminogen was 
identified among the urinary impurities; plasminogen has been identified as a 
binding protein for disease-associated prion protein [17].
Both hMG and hMG-HP were tested using 2D gel electrophoresis. The results 
confirmed that the two hMGs tested were less pure than hMG-HP and contained a 
large number of total proteins (mainly represented by urinary impurities). 
Non-gonadotropin proteins present in hMG-HP products were identified by MS, 
resulting in 34 co-purified contaminants. Only gonadotropin proteins were seen 
in all the recombinant preparations analyzed by 2D gel electrophoresis. Using 
this 2D-gel/LC-MS/MS proteomic workflow, prion proteins were identified only in 
u-hCG and not in hMG-HP preparations. In parallel, a targeted proteomic approach 
(LC-SRM) was developed to detect human prion proteins which are sensitive to 
proteases. The method was optimized to provide quantitative data in each 
container of product. This is the most sensitive MS-based quantification 
technique currently available with a limit of detection in the low femtomolar 
range. This approach for prion protein detection, identification and 
quantification was used on all gonadotropin pharmaceutical preparations included 
in our study, including both urinary (hCG, hMG, hMG-HP) and recombinant 
products.
All urine-derived preparations tested, produced by different manufacturers, 
showed the presence of human prion proteins in varying amounts. These findings 
demonstrate that the purification processes for different urine-derived 
preparations are unable to remove prion proteins from the source material and 
that the process controls employed do not permit the identification of this 
contaminant.
Do the prion protein peptides detected in this study originate from 
infectious prions? Preparation of tryptic peptides is preceded by solubilization 
in 8M urea, which is adequate to disaggregate and denature the 
disease-associated isoform of the prion protein rendering it susceptible to 
trypsin digestion. It is also clear that native and diseased isoforms of the 
prion protein share affinity for chromatography substrates utilized to purify 
peptide hormones [3]. Finally, infectious prions can range down in size to 
oligomers of a few dozen prion protein molecules [18], which would be 
undetectable by existing biochemical methodologies including MS methods employed 
in this study.
Although no cases of human prion disease due to the use of urinary 
gonadotropins have been recognized to date, the epidemiological signal for 
transmission may be difficult to detect. Each year, more than 300,000 young 
women in the US and Canada are prescribed urine-derived gonadotropins for 
infertility. Although the Food and Drug Administration and Health Canada once 
considered these products to be in the lowest category of risk for prion disease 
transmission, the discovery of full infectivity in the urine of nephritic 
scrapie-infected mice in 2005 led to new requirements for product labeling and a 
review of donor procedures and manufacturing processes. Additional recent 
findings suggest that urinary prion excretion can occur without renal pathology 
[6], [7]. These results warrant a reassessment as to whether the risks of 
urine-derived fertility products could now outweigh their benefits, particularly 
considering the availability of recombinant products that do not require human 
urine as a substrate.
Although urinary gonadotropins have been previously characterized as safe 
[19], [20], this opinion may be overly optimistic in view of the present 
findings, supported by results from other recently published studies. Notably, 
blood products were once also considered ‘safe’, based on the lack of detectable 
prions in vCJD using an inadequately sensitive mouse bioassay [21]. In line with 
recent published studies, the 2010 updated World Health Organization tables on 
‘Tissue infectivity distribution in transmissible spongiform encephalopathy’ 
moved urine from the category of ‘Tissues with no detectable infectivity’ to the 
category of ‘Lower-infectivity tissue’ (the latter category includes blood) 
[22].
Current urine collection systems pool the urine of thousands of donors and, 
unlike the blood collection system, do not allow for donor tracing. There is 
also no mechanism of ensuring that the designated donor is actually the one who 
provides the urine, as donation is normally done at home. However, even if donor 
management and tracing were flawless, the fact that prionuria may exist well 
before the onset of clinically overt prion disease, without being detectable by 
current methods, remains a cause for concern. Furthermore, the now indisputable 
detection of prions in urine of experimental animals, the lack of a species 
barrier for human-to-human transmission, the relative efficiency of the 
intramuscular injection route for prion transmission, and the young age of 
fertility drug recipients all support application of the ‘precautionary 
principle’ for urinary derived pharmaceuticals. As risk management paradigms 
shift towards more proactive approaches intended to ‘anticipate and prevent’ 
emerging risks [23]–[26], a careful examination of the risk of transmission of 
human prion disease through the use of urine-derived hormones and peptides would 
appear to be warranted.
Friday, March 25, 2011
Detection of Prion Protein in Urine-Derived Injectable Fertility Products 
by a Targeted Proteomic Approach
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. 
Monday, June 17, 2013 
Early detection of chronic wasting disease prions in urine of 
pre-symptomatic deer by real-time quaking-induced conversion assay 
Thursday, April 12, 2012
Health professions and risk of sporadic Creutzfeldt–Jakob disease, 1965 to 
2010 
Eurosurveillance, Volume 17, Issue 15, 12 April 2012 
Research articles
Tuesday, May 28, 2013 
Late-in-life surgery associated with Creutzfeldt-Jakob disease: a 
methodological outline for evidence-based guidance 
Sunday, June 9, 2013 
TSEAC March 14, 2013: Transmissible Spongiform Encephalopathies Advisory 
Committee Meeting Webcast 
Tuesday, May 21, 2013 
CJD, TSE, PRION, BLOOD Abstracts of the 23rd Regional Congress of the 
International Society of Blood Transfusion, Amsterdam, The Netherlands, June 
2-5, 2013 
Tuesday, March 5, 2013 
Use of Materials Derived From Cattle in Human Food and Cosmetics; Reopening 
of the Comment Period FDA-2004-N-0188-0051 (TSS SUBMISSION) 
FDA believes current regulation protects the public from BSE but reopens 
comment period due to new studies 
Thursday, October 25, 2012 
Current limitations about the cleaning of luminal endoscopes and TSE prion 
risk factors there from 
Article in Press 
Saturday, January 16, 2010 
Evidence For CJD TSE Transmission Via Endoscopes 1-24-3 re-Singeltary to 
Bramble et al 
Evidence For CJD/TSE Transmission Via Endoscopes 
From Terry S. Singletary, Sr flounder@wt.net 1-24-3 
Tuesday, July 31, 2012 
11 patients may have been exposed to fatal disease Creutzfeldt-Jakob 
Disease CJD Greenville Memorial Hospital 
Thursday, August 02, 2012 
CJD case in Saint John prompts letter to patients Canada CJD case in Saint 
John prompts letter to patients 
Saturday, February 12, 2011 
Another Pathologists dies from CJD, another potential occupational death ? 
another happenstance of bad luck, a spontaneous event from nothing, or 
friendly fire ??? 
Thursday, July 08, 2010 
GLOBAL CLUSTERS OF CREUTZFELDT JAKOB DISEASE - A REVIEW 2010 
Sunday, May 10, 2009 
Meeting of the Transmissible Spongiform Encephalopathies Committee On June 
12, 2009 (Singeltary submission) 
Thursday, January 29, 2009 
Medical Procedures and Risk for Sporadic Creutzfeldt-Jakob Disease, Japan, 
1999-2008 (WARNING TO Neurosurgeons and Ophthalmologists) Volume 15, Number 
2-February 2009 Research 
Wednesday, August 20, 2008 
Tonometer disinfection practice in the United Kingdom: A national survey 
Tuesday, August 12, 2008 
Biosafety in Microbiological and Biomedical Laboratories Fifth Edition 2007 
(occupational exposure to prion diseases) 
Monday, December 31, 2007 
Risk Assessment of Transmission of Sporadic Creutzfeldt-Jakob Disease in 
Endodontic Practice in Absence of Adequate Prion Inactivation 
Subject: CJD: update for dental staff 
Date: November 12, 2006 at 3:25 pm PST 
1: Dent Update. 2006 Oct;33(8):454-6, 458-60. 
CJD: update for dental staff. 
Friday, July 19, 2013 
Beaumont Hospital in Dublin assessing patients for CJD 
Saturday, September 21, 2013 
CJD CONFIRMED in patient at New Hampshire Department of Health and Human 
Services (DHHS), Catholic Medical Center (CMC), and the Manchester Health 
Department (MHD) 
Thursday, September 26, 2013 
Minimise transmission risk of CJD and vCJD in healthcare settings Guidance 
Wednesday, June 19, 2013 
Spreading of tau pathology in Alzheimer's disease by cell-to-cell 
transmission 
MAD COW TESTING ONLY CATCHES SOME MAD COWS 
SPREADING IT ALL AROUND
Saturday, October 19, 2013 
***A comparative study of modified confirmatory techniques and additional 
immuno-based methods for non-conclusive autolytic Bovine spongiform 
encephalopathy cases 
Wednesday, September 25, 2013 
Inspections, Compliance, Enforcement, and Criminal Investigations BSE TSE 
PRION 2013 
Wednesday, October 09, 2013 
WHY THE UKBSEnvCJD ONLY THEORY IS SO POPULAR IN IT'S FALLACY, £41,078,281 
in compensation REVISED
Thursday, October 10, 2013 
CJD REPORT 1994 increased risk for consumption of veal and venison and lamb 
Monday, October 14, 2013 
Researchers estimate one in 2,000 people in the UK carry variant CJD 
proteins 
Friday, August 16, 2013 
*** Creutzfeldt-Jakob disease (CJD) biannual update August 2013 U.K. and 
Contaminated blood products induce a highly atypical prion disease devoid of 
PrPres in primates 
WHAT about the sporadic CJD TSE proteins ?
WE now know that some cases of sporadic CJD are linked to atypical BSE and 
atypical Scrapie, so why are not MORE concerned about the sporadic CJD, and all 
it’s sub-types $$$ 
Sunday, August 11, 2013 
Creutzfeldt-Jakob Disease CJD cases rising North America updated report 
August 2013 
Creutzfeldt-Jakob Disease CJD cases rising North America with Canada seeing 
an extreme increase of 48% between 2008 and 2010 
Sunday, July 21, 
2013 
Welsh Government and Food Standards Agency Wales 
Joint Public Consultation on the Proposed Transmissible Spongiform 
Encephalopathies (Wales) Regulations 2013 Singeltary Submission WG18417 
Sunday, October 13, 2013 
CJD TSE Prion Disease Cases in Texas by Year, 2003-2012
Tuesday, September 24, 2013 
NORDION (US), INC., AND BIOAXONE BIOSCIENCES, INC., Settles $90M Mad Cow 
TSE prion Contamination Suit Cethrin(R) 
Case 0:12-cv-60739-RNS Document 1 Entered on FLSD Docket 04/26/2012 Page 1 
of 15 
Monday, October 21, 2013 
WTO Mad cow disease (No 193) 
Saturday, July 6, 2013 
Small Ruminant Nor98 Prions Share Biochemical 
Features with Human Gerstmann-Sträussler-Scheinker Disease and Variably 
Protease-Sensitive Prionopathy 
Research Article 
Thursday, March 29, 2012
atypical Nor-98 Scrapie has spread from coast to 
coast in the USA 2012 
NIAA Annual Conference April 11-14, 2011San Antonio, 
Texas
Monday, April 25, 2011
Experimental Oral Transmission of Atypical Scrapie to 
Sheep
Volume 17, Number 5-May 2011 However, work with 
transgenic mice has demonstrated the potential susceptibility of pigs, with the 
disturbing finding that the biochemical properties of the resulting PrPSc have 
changed on transmission (40). 
Monday, December 14, 2009
Similarities between Forms of Sheep Scrapie and 
Creutzfeldt-Jakob Disease Are Encoded by Distinct Prion Types
(hmmm, this is getting interesting now...TSS)
Sporadic CJD type 1 and atypical/ Nor98 scrapie are 
characterized by fine (reticular) deposits,
see also ;
All of the Heidenhain variants were of the 
methionine/ methionine type 1 molecular subtype.
see full text ;
Monday, December 14, 2009
Similarities between Forms of Sheep Scrapie and 
Creutzfeldt-Jakob Disease Are Encoded by Distinct Prion Types
Friday, March 09, 2012 
Experimental H-type and L-type bovine spongiform 
encephalopathy in cattle: observation of two clinical syndromes and diagnostic 
challenges 
Research article 
Thursday, June 23, 2011 
Experimental H-type bovine spongiform encephalopathy 
characterized by plaques and glial- and stellate-type prion protein deposits 
Tuesday, August 22, 2006
BSE ATYPICAL TEXAS AND ALABAMA UPDATE JANUARY 20, 2007 
 2009 UPDATE ON ALABAMA MAD COW FOUND IN 2006
http://bse-atypical.blogspot.com/2006/08/bse-atypical-texas-and-alabama-update.html 
I ask Professor Kong ; 
Thursday, December 04, 2008 3:37 PM 
Subject: RE: re--Chronic Wating Disease (CWD) and Bovine Spongiform 
Encephalopathies (BSE): Public Health Risk Assessment
''IS the h-BSE more virulent than typical BSE as well, or the same as cBSE, 
or less virulent than cBSE? just curious.....''
Professor Kong reply ; 
.....snip 
''As to the H-BSE, we do not have sufficient data to say one way or 
another, but we have found that H-BSE can infect humans. I hope we could publish 
these data once the study is complete. Thanks for your interest.''
Best regards,
Qingzhong Kong, PhD Associate Professor Department of Pathology Case 
Western Reserve University Cleveland, OH 44106 USA
END...TSS 
Thursday, December 04, 2008 2:37 PM
"we have found that H-BSE can infect humans."
personal communication with Professor Kong. ...TSS 
BSE-H is also transmissible in our humanized Tg mice. 
The possibility of more than two atypical BSE strains will be discussed. 
Supported by NINDS NS052319, NIA AG14359, and NIH AI 77774. 
please see below from PRION2013 ; 
*** This study imply the possibility that the novel BSE prions with high 
virulence in cattle will be emerged during intraspecies transmission. 
AD.56: The emergence of novel BSE prions by serial passages of H-type BSE 
in bovinized mice 
Kentaro Masujin, Naoko Tabeta, Ritsuko Miwa, Kohtaro Miyazawa, Hiroyuki 
Okada, Shirou Mohri and Takashi Yokoyama 
National Institute of Animal Health; Tsukuba, Japan 
H-type bovine spongiform encephalopathy (BSE) is an atypical form of BSE, 
and has been detected in several European countries, and North America. 
Transmission studies of H-type BSE led to the emergence of the classical BSE 
(C-BSE) phenotypes during passages in inbred wild type and bovinized 
PrP-overexpressing transgenic mice. In this study, we conducted serial passages 
of Canadian H-type BSE isolate in bovinized PrP-overexpressing transgenic mice 
(TgBoPrP). H-type BSE isolate was transmitted to TgBoPrP with incubation periods 
of 320 ± 12.2 d at primary passage. The incubation period of 2nd and 3rd passage 
were constant (~= 220 d), no clear differences were observed in their biological 
and biochemical properties. However, at the forth passage, 2 different BSE 
phenotypes were confirmed; one is shorter survival times (109 ± 4 d) and the 
other is longer survival times. TgBoPrP mice with longer incubation period 
showed the H-type phenotype of PrPsc profile and pathology. However, those of 
shorter incubation period were different phenotypes from previously existed BSE 
prions (C-BSE, L-type BSE, and H-type BSE). *** This study imply the possibility 
that the novel BSE prions with high virulence in cattle will be emerged during 
intraspecies transmission. 
please see ; 
Thursday, August 15, 2013 
The emergence of novel BSE prions by serial passages of H-type BSE in 
bovinized mice 
Wednesday, September 25, 2013 
Presence of subclinical infection in gene-targeted human prion protein 
transgenic mice exposed to atypical BSE 
*** TESTING MAD COWS ONLY CONFIRMS _SOME_ MAD COWS ***
HOW MANY WERE MISSED, AND CONSUMBED ???
Saturday, October 19, 2013 
A comparative study of modified confirmatory techniques and additional 
immuno-based methods for non-conclusive autolytic Bovine spongiform 
encephalopathy cases 
Friday, October 25, 2013 
*** UK FSA TSE BSE Board meeting agenda: 5 November 2013 ***
P03.141
Aspects of the Cerebellar Neuropathology in 
Nor98
Gavier-Widén, D1; Benestad, SL2; Ottander, L1; 
Westergren, E1 1National Veterinary Insitute, Sweden; 2National Veterinary 
Institute,
Norway Nor98 is a prion disease of old sheep and 
goats. This atypical form of scrapie was first described in Norway in 1998. 
Several features of Nor98 were shown to be different from classical scrapie 
including the distribution of disease associated prion protein (PrPd) 
accumulation in the brain. The cerebellum is generally the most affected brain 
area in Nor98. The study here presented aimed at adding information on the 
neuropathology in the cerebellum of Nor98 naturally affected sheep of various 
genotypes in Sweden and Norway. A panel of histochemical and immunohistochemical 
(IHC) stainings such as IHC for PrPd, synaptophysin, glial fibrillary acidic 
protein, amyloid, and cell markers for phagocytic cells were conducted. The type 
of histological lesions and tissue reactions were evaluated. The types of PrPd 
deposition were characterized. The cerebellar cortex was regularly affected, 
even though there was a variation in the severity of the lesions from case to 
case. Neuropil vacuolation was more marked in the molecular layer, but affected 
also the granular cell layer. There was a loss of granule cells. Punctate 
deposition of PrPd was characteristic. It was morphologically and in 
distribution identical with that of synaptophysin, suggesting that PrPd 
accumulates in the synaptic structures. PrPd was also observed in the granule 
cell layer and in the white matter. The pathology features of Nor98 in the 
cerebellum of the affected sheep showed similarities with those of sporadic 
Creutzfeldt-Jakob disease in humans.
***The pathology features of Nor98 in the cerebellum 
of the affected sheep showed similarities with those of sporadic 
Creutzfeldt-Jakob disease in humans.
PR-26
NOR98 SHOWS MOLECULAR FEATURES REMINISCENT OF 
GSS
R. Nonno1, E. Esposito1, G. Vaccari1, E. Bandino2, M. 
Conte1, B. Chiappini1, S. Marcon1, M. Di Bari1, S.L. Benestad3, U. Agrimi1 1 
Istituto Superiore di Sanità, Department of Food Safety and Veterinary Public 
Health, Rome, Italy (romolo.nonno@iss.it); 2 Istituto Zooprofilattico della 
Sardegna, Sassari, Italy; 3 National Veterinary Institute, Department of 
Pathology, Oslo, Norway
Molecular variants of PrPSc are being increasingly 
investigated in sheep scrapie and are generally referred to as "atypical" 
scrapie, as opposed to "classical scrapie". Among the atypical group, Nor98 
seems to be the best identified. We studied the molecular properties of Italian 
and Norwegian Nor98 samples by WB analysis of brain homogenates, either 
untreated, digested with different concentrations of proteinase K, or subjected 
to enzymatic deglycosylation. The identity of PrP fragments was inferred by 
means of antibodies spanning the full PrP sequence. We found that undigested 
brain homogenates contain a Nor98-specific PrP fragment migrating at 11 kDa 
(PrP11), truncated at both the C-terminus and the N-terminus, and not 
N-glycosylated. After mild PK digestion, Nor98 displayed full-length PrP 
(FL-PrP) and N-glycosylated C-terminal fragments (CTF), along with increased 
levels of PrP11. Proteinase K digestion curves (0,006-6,4 mg/ml) showed that 
FL-PrP and CTF are mainly digested above 0,01 mg/ml, while PrP11 is not entirely 
digested even at the highest concentrations, similarly to PrP27-30 associated 
with classical scrapie. Above 0,2 mg/ml PK, most Nor98 samples showed only PrP11 
and a fragment of 17 kDa with the same properties of PrP11, that was tentatively 
identified as a dimer of PrP11. Detergent solubility studies showed that PrP11 
is insoluble in 2% sodium laurylsorcosine and is mainly produced from 
detergentsoluble, full-length PrPSc. Furthermore, among Italian scrapie 
isolates, we found that a sample with molecular and pathological properties 
consistent with Nor98 showed plaque-like deposits of PrPSc in the thalamus when 
the brain was analysed by PrPSc immunohistochemistry. Taken together, our 
results show that the distinctive pathological feature of Nor98 is a PrP 
fragment spanning amino acids ~ 90-155. This fragment is produced by successive 
N-terminal and C-terminal cleavages from a full-length and largely 
detergent-soluble PrPSc, is produced in vivo and is extremely resistant to PK 
digestion.
*** Intriguingly, these conclusions suggest that some 
pathological features of Nor98 are reminiscent of Gerstmann-Sträussler-Scheinker 
disease.
119
A newly identified type of scrapie agent can 
naturally infect sheep with resistant PrP genotypes
Annick Le Dur*,?, Vincent Béringue*,?, Olivier 
Andréoletti?, Fabienne Reine*, Thanh Lan Laï*, Thierry Baron§, Bjørn Bratberg¶, 
Jean-Luc Vilotte?, Pierre Sarradin**, Sylvie L. Benestad¶, and Hubert Laude*,?? 
+Author Affiliations
*Virologie Immunologie Moléculaires and ?Génétique 
Biochimique et Cytogénétique, Institut National de la Recherche Agronomique, 
78350 Jouy-en-Josas, France; ?Unité Mixte de Recherche, Institut National de la 
Recherche Agronomique-Ecole Nationale Vétérinaire de Toulouse, Interactions Hôte 
Agent Pathogène, 31066 Toulouse, France; §Agence Française de Sécurité Sanitaire 
des Aliments, Unité Agents Transmissibles Non Conventionnels, 69364 Lyon, 
France; **Pathologie Infectieuse et Immunologie, Institut National de la 
Recherche Agronomique, 37380 Nouzilly, France; and ¶Department of Pathology, 
National Veterinary Institute, 0033 Oslo, Norway
***Edited by Stanley B. Prusiner, University of 
California, San Francisco, CA (received for review March 21, 2005)
Abstract Scrapie in small ruminants belongs to 
transmissible spongiform encephalopathies (TSEs), or prion diseases, a family of 
fatal neurodegenerative disorders that affect humans and animals and can 
transmit within and between species by ingestion or inoculation. Conversion of 
the host-encoded prion protein (PrP), normal cellular PrP (PrPc), into a 
misfolded form, abnormal PrP (PrPSc), plays a key role in TSE transmission and 
pathogenesis. The intensified surveillance of scrapie in the European Union, 
together with the improvement of PrPSc detection techniques, has led to the 
discovery of a growing number of so-called atypical scrapie cases. These include 
clinical Nor98 cases first identified in Norwegian sheep on the basis of unusual 
pathological and PrPSc molecular features and "cases" that produced discordant 
responses in the rapid tests currently applied to the large-scale random 
screening of slaughtered or fallen animals. Worryingly, a substantial proportion 
of such cases involved sheep with PrP genotypes known until now to confer 
natural resistance to conventional scrapie. Here we report that both Nor98 and 
discordant cases, including three sheep homozygous for the resistant PrPARR 
allele (A136R154R171), efficiently transmitted the disease to transgenic mice 
expressing ovine PrP, and that they shared unique biological and biochemical 
features upon propagation in mice. *** These observations support the view that 
a truly infectious TSE agent, unrecognized until recently, infects sheep and 
goat flocks and may have important implications in terms of scrapie control and 
public health.
Monday, December 1, 2008
When Atypical Scrapie cross species barriers
Authors
Andreoletti O., Herva M. H., Cassard H., Espinosa J. 
C., Lacroux C., Simon S., Padilla D., Benestad S. L., Lantier F., Schelcher F., 
Grassi J., Torres, J. M., UMR INRA ENVT 1225, Ecole Nationale Veterinaire de 
Toulouse.France; ICISA-INlA, Madrid, Spain; CEA, IBiTec-5, DSV, CEA/Saclay, Gif 
sur Yvette cedex, France; National Veterinary Institute, Postboks 750 Sentrum, 
0106 Oslo, Norway, INRA IASP, Centre INRA de Tours, 3738O Nouzilly, 
France.
Content
Atypical scrapie is a TSE occurring in small 
ruminants and harbouring peculiar clinical, epidemiological and biochemical 
properties. Currently this form of disease is identified in a large number of 
countries. In this study we report the transmission of an atypical scrapie 
isolate through different species barriers as modeled by transgenic mice (Tg) 
expressing different species PRP sequence.
The donor isolate was collected in 1995 in a French 
commercial sheep flock. inoculation into AHQ/AHQ sheep induced a disease which 
had all neuro-pathological and biochemical characteristics of atypical scrapie. 
Transmitted into Transgenic mice expressing either ovine or PrPc, the isolate 
retained all the described characteristics of atypical scrapie.
Surprisingly the TSE agent characteristics were 
dramatically different v/hen passaged into Tg bovine mice. The recovered TSE 
agent had biological and biochemical characteristics similar to those of 
atypical BSE L in the same mouse model. Moreover, whereas no other TSE agent 
than BSE were shown to transmit into Tg porcine mice, atypical scrapie was able 
to develop into this model, albeit with low attack rate on first passage.
Furthermore, after adaptation in the porcine mouse 
model this prion showed similar biological and biochemical characteristics than 
BSE adapted to this porcine mouse model. Altogether these data indicate.
(i) the unsuspected potential abilities of atypical 
scrapie to cross species barriers
(ii) the possible capacity of this agent to acquire 
new characteristics when crossing species barrier
These findings raise some interrogation on the 
concept of TSE strain and on the origin of the diversity of the TSE agents and 
could have consequences on field TSE control measures.
Friday, February 11, 2011 
Atypical/Nor98 Scrapie Infectivity in Sheep 
Peripheral Tissues 
RESEARCH 
Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 
17, No. 5, May 2011 
Experimental Oral Transmission of Atypical Scrapie to 
Sheep 
Marion M. Simmons, S. Jo Moore,1 Timm Konold, Lisa 
Thurston, Linda A. Terry, Leigh Thorne, Richard Lockey, Chris Vickery, Stephen 
A.C. Hawkins, Melanie J. Chaplin, and John Spiropoulos 
To investigate the possibility of oral transmission 
of atypical scrapie in sheep and determine the distribution of infectivity in 
the animals’ peripheral tissues, we challenged neonatal lambs orally with 
atypical scrapie; they were then killed at 12 or 24 months. Screening test 
results were negative for disease-specifi c prion protein in all but 2 
recipients; they had positive results for examination of brain, but negative for 
peripheral tissues. Infectivity of brain, distal ileum, and spleen from all 
animals was assessed in mouse bioassays; positive results were obtained from 
tissues that had negative results on screening. These fi ndings demonstrate that 
atypical scrapie can be transmitted orally and indicate that it has the 
potential for natural transmission and iatrogenic spread through animal feed. 
Detection of infectivity in tissues negative by current surveillance methods 
indicates that diagnostic sensitivity is suboptimal for atypical scrapie, and 
potentially infectious material may be able to pass into the human food chain. 
SNIP... 
Although we do not have epidemiologic evidence that 
supports the effi cient spread of disease in the fi eld, these data imply that 
disease is potentially transmissible under fi eld situations and that spread 
through animal feed may be possible if the current feed restrictions were to be 
relaxed. Additionally, almost no data are available on the potential for 
atypical scrapie to transmit to other food animal species, certainly by the oral 
route. However, work with transgenic mice has demonstrated the potential 
susceptibility of pigs, with the disturbing fi nding that the biochemical 
properties of the resulting PrPSc have changed on transmission (40). The 
implications of this observation for subsequent transmission and host target 
range are currently unknown. 
How reassuring is this absence of detectable PrPSc 
from a public health perspective? The bioassays performed in this study are not 
titrations, so the infectious load of the positive gut tissues cannot be 
quantifi ed, although infectivity has been shown unequivocally. No experimental 
data are currently available on the zoonotic potential of atypical scrapie, 
either through experimental challenge of humanized mice or any meaningful 
epidemiologic correlation with human forms of TSE. However, the detection of 
infectivity in the distal ileum of animals as young as 12 months, in which all 
the tissues tested were negative for PrPSc by the currently available screening 
and confi rmatory diagnostic tests, indicates that the diagnostic sensitivity of 
current surveillance methods is suboptimal for detecting atypical scrapie and 
that potentially infectious material may be able to pass into the human food 
chain undetected. 
Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 
17, No. 5, May 2011 
Saturday, October 
19, 2013 
ACA Council Meets 
to Endorse Several Proposed USAHA Resolutions (CWD TSE PRION DISEASE) 
*** 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. 
TSS

 
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