Tuesday, November 11, 2008

Transmission of atypical bovine prions to mice transgenic for human prion protein

DOI: 10.3201/eid1412.080941

Suggested citation for this article: Béringue V, Herzog L, Reine F, Le Dur A, Casalone C, Vilotte J-L, et al.

Transmission of atypical bovine prions to mice transgenic for human prion protein.

Emerg Infect Dis. 2008 Dec; [Epub ahead of print]

Transmission of Atypical Bovine Prions to Mice Transgenic for Human Prion Protein

Vincent Béringue, Laëtitia Herzog, Fabienne Reine, Annick Le Dur, Cristina Casalone, Jean-Luc Vilotte, and Hubert Laude Author affiliations: Institut National de la Recherche Agronomique, Jouy-en-Josas, France (V. Béringue, L. Herzog, F. Reine, A. Le Dur, H. Laude, J.-L. Vilotte); and Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Turin, Italy (C. Casalone)

To assess risk for cattle-to-human transmission of prions that cause uncommon forms of bovine spongiform encephalopathy (BSE), we inoculated mice expressing human PrP Met129 with field isolates. Unlike classical BSE agent, L-type prions appeared to propagate in these mice with no obvious transmission barrier. H-type prions failed to infect the mice.

The epizootic of bovine spongiform encephalopathy (BSE) is under control in European countries >20 years after the first cases were diagnosed in the United Kingdom. Thus far, BSE is the only animal prion disease known to have been transmitted to humans, leading to a variant form of Creutzfeldt-Jakob disease (vCJD) (1). The large-scale testing of livestock nervous tissues for the presence of protease-resistant prion protein (PrPres) has enabled assessment of BSE prevalence and exclusion of BSE-infected animals from human food (2). This active surveillance has led to the recognition of 2 variant PrPres molecular signatures, termed H-type and L-type BSE. They differ from that of classical BSE by having protease-resistant fragments of a higher (H) or a slightly lower (L) molecular mass, respectively, and different patterns of glycosylation (3–5). Both types have been detected worldwide as rare cases in older animals, at a low prevalence consistent with the possibility of sporadic forms of prion diseases in cattle (6).

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Their experimental transmission to mice transgenic for bovine PrP demonstrated the infectious nature of such cases and the existence of distinct prion strains in cattle (5,7–9). Like the classical BSE agent, H- and L-type prions can propagate in heterologous species (7–11). Thus, both agents are transmissible to transgenic mice expressing ovine PrP (VRQ allele). Although H-type molecular properties are conserved on these mice (9), L-type prions acquire molecular and neuropathologic phenotypic traits undistinguishable from BSE or BSE-related agents that have followed the same transmission history (7). Similar findings have been reported in wild-type mice (8). An understanding of the transmission properties of these newly recognized prions when confronted with the human PrP sequence is needed. In a previous study, we measured kinetics of PrPres deposition in the brain to show that L-type prions replicate faster than BSE prions in experimentally inoculated mice that express human PrP (7). In a similar mouse model, the L-type agent (alternatively named BASE) was also shown to produce overt disease with an attack rate of ˜30% (12). However, no strict comparison with BSE agent has been attempted. As regards the H-type agent, its potential virulence for mice that express human PrP Met129 remains to be assessed. We now report comparative transmission data for these atypical and classical BSE prions.

The Study

The bovine isolates used in this study have been previously described; they all exhibited high infectivity levels in bovine PrP mice (4,7,9). The equivalent of 2 mg of infected bovine brain tissue was injected intracerebrally into tg650 mice. This line of mice overexpresses (˜6-fold) human PrP with methionine at codon 129 (Met129) on a Zurich mouse PrP null background and has been shown to be fully susceptible to vCJD agent (13). The resulting transmission data available to date are summarized in the Table. The primary transmission of classical BSE isolates was inefficient as judged by the absence of clear neurologic signs and by Western blot detection of PrPres in the brain of only 4/25 inoculated mice. The PrPres banding pattern was essentially similar to that of vCJD (low molecular mass fragments and predominance of diglycoform species;

Figure 1). Secondary passages were performed by using PrPres-negative or PrPres-positive individual mouse brains. Every time brain homogenate from an aged mouse (>630 days of age) was

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inoculated, transmission was observed in >80% of the mice, as determined by clinical signs and PrPres accumulation. The mean survival time was ˜600 days (Table and data not shown). By the third passage, mean survival time approached ˜500 days, as is usually observed with vCJD cases (Table; 13). The vCJD-like PrPres profile was conserved in all 50 positive brains analyzed (Figure 1). Transmission of L-type isolates to tg650 mice produced markedly different results. First, the 4 L-type isolates induced neurologic disease in almost all mice; survival times averaged 600–700 days (Table). Second, PrPres accumulated in the brain of all 33 mice analyzed. The molecular profile was distinct from BSE or vCJD; prominent monoglycosylated PrPres species resembled those found in cattle (Figure 1). Third, we found neither shortening of the survival time on subpassage (1 isolate tested, 2 brains) (Table; data not shown) nor change in PrPres profile

(Figure 1).

In sharp contrast, BSE H-type isolates failed to transmit disease to or even infect tg650 mice. None of the inoculated mice had a detectable level of PrPres in the brain (22 analyzed). Secondary passages were performed with brains of mice that died at various time points. All inoculated mice survived, and none showed a PrPres signal in the brain (15 mice analyzed). To further compare the behavior of the 3 bovine prions in tg650 mice, we examined the regional distribution and intensity of PrPres deposition in the brain (14). Histoblot analyses (3 brains per infection) were performed on primary (L-type, H-type) and secondary passages (L-type, H-type, BSE). As shown in Figure 2, L-type and BSE agents showed clear differences according to both the aspect and localization of the PrP deposits. Granular PrP deposits were scattered throughout the brain with BSE, as has been previously observed with vCJD (13). The ventral nuclei of the thalamus, cerebral cortex, oriens layer of the hippocampus, and raphe and tegmentum nuclei of the brain stem were strongly stained. With BSE-L, the staining was finer and essentially confined to the habenular, geniculate, and dorsal nuclei of the thalamus; the lateral hypothalamus; the lacunosom moleculare layer of the hippocampus; the superficial gray layer of the superior colliculus; and the raphe nuclei of the brain stem. Finally, PrPres could not be detected on brain sections from mice inoculated with H-type isolates (data not shown), thus confirming the Western blot data.

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We found that atypical L-type bovine prions can propagate in human PrP transgenic mice with no significant transmission barrier. Lack of a barrier is supported by the 100% attack rate, the absence of reduction of incubation time on secondary passage, and the conservation of PrPres electrophoretic profile. In comparison, transmission of classical BSE agent to the same mice showed a substantial barrier. Indeed, 3 passages were necessary to reach a degree of virulence comparable to that of vCJD agent in these mice (13), which likely reflects progressive adaptation of the agent to its new host. At variance with the successful transmission of classical BSE and L-type agents, H-type agent failed to infect tg650 mice. These mice overexpress human PrP and were inoculated intracranially with a low dilution inoculum (10% homogenate). Therefore, this result supports the view that the transmission barrier of BSE-H from cattle to humans might be quite robust. It also illustrates the primacy of the strain over PrP sequence matching for cross-species transmission of prions (15). Extrapolation of our data raises the theoretical possibility that the zoonotic risk associated with BSE-L prions might be higher than that associated with classical BSE, at least for humans carrying the Met129 PrP allele. This information underlines the need for more intensive investigations, in particular regarding the tissue tropism of this agent. Its ability to colonize lymphoid tissues is a potential, key factor for a successful transmission by peripheral route. This issue is currently being explored in the tg650 mice. Although recent data in humanized mice suggested that BSE-L agent is likely to be lymphotropic (12), preliminary observations in our model suggested that its ability to colonize such tissues is comparatively much lower than that of classical BSE agent.


We thank A.G. Biacabe and T. Baron for providing the BSE-L and BSE-H isolates from France and the staff from Unité Expérimentale Animalerie Rongeurs of the Institut National de la Recherche Agronomique, Jouy-en-Josas, for excellent mouse care. This work was supported by a joint grant from Institut National de la Recherche Agronomique–Agence Française de Sécurité Sanitaire des Aliments and by the European Network of Excellence NeuroPrion. Page 4 of 8 Dr Béringue is a senior scientist at the Institut National de la Recherche Agronomique in Jouy-en-Josas. His primary research interests include the diversity, pathogenesis, and potential of interspecies transmission of animal and human prions. References 1. Wadsworth JD, Collinge J. Update on human prion disease. Biochim Biophys Acta. 2007;1772:598–609. 2. Grassi J, Creminon C, Frobert Y, Fretier P, Turbica I, Rezaei H, et al. Specific determination of the proteinase K-resistant form of the prion protein using two-site immunometric assays. Application to the post-mortem diagnosis of BSE. Arch Virol Suppl. 2000;16:197–205. PubMed 3. Biacabe AG, Laplanche JL, Ryder S, Baron T. Distinct molecular phenotypes in bovine prion diseases. EMBO Rep. 2004;5:110–5. PubMed DOI: 10.1038/sj.embor.7400054 4. Casalone C, Zanusso G, Acutis P, Ferrari S, Capucci L, Tagliavini F, et al. Identification of a second bovine amyloidotic spongiform encephalopathy: molecular similarities with sporadic Creutzfeldt-Jakob disease. Proc Natl Acad Sci U S A. 2004;101:3065–70. PubMed DOI: 10.1073/pnas.0305777101 5. Buschmann A, Gretzschel A, Biacabe AG, Schiebel K, Corona C, Hoffmann C, et al. Atypical BSE in Germany—proof of transmissibility and biochemical characterization. Vet Microbiol. 2006;117:103–16. PubMed DOI: 10.1016/j.vetmic.2006.06.016 6. Biacabe AG, Morignat E, Vulin J, Calavas D, Baron TG. Atypical bovine spongiform encephalopathies, France, 2001–2007. Emerg Infect Dis. 2008;14:298–300. PubMed DOI: 10.3201/eid1402.071141 7. Beringue V, Andreoletti O, Le Dur A, Essalmani R, Vilotte JL, Lacroux C, et al. A bovine prion acquires an epidemic bovine spongiform encephalopathy strain-like phenotype on interspecies transmission. J Neurosci. 2007;27:6965–71. PubMed DOI: 10.1523/JNEUROSCI.0693-07.2007 8. Capobianco R, Casalone C, Suardi S, Mangieri M, Miccolo C, Limido L, et al. Conversion of the BASE prion strain into the BSE strain: the origin of BSE? PLoS Pathog. 2007;3:e31. PubMed DOI: 10.1371/journal.ppat.0030031 9. Béringue V, Bencsik A, Le Dur A, Reine F, Lai TL, Chenais N, et al. Isolation from cattle of a prion strain distinct from that causing bovine spongiform encephalopathy. PLoS Pathog. 2006;2:e112. PubMed DOI: 10.1371/journal.ppat.0020112

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10. Baron T, Bencsik A, Biacabe AG, Morignat E, Bessen RA. Phenotypic similarity of transmissible mink encephalopathy in cattle and L-type bovine spongiform encephalopathy in a mouse model. Emerg Infect Dis. 2007;13:1887–94. PubMed 11. Baron TG, Biacabe AG, Bencsik A, Langeveld JP. Transmission of new bovine prion to mice. Emerg Infect Dis. 2006;12:1125–8. PubMed 12. Kong Q, Zheng M, Casalone C, Qing L, Huang S, Chakraborty B, et al. Evaluation of the human transmission risk of an atypical bovine spongiform encephalopathy prion strain. J Virol. 2008;82:3697–701. PubMed DOI: 10.1128/JVI.02561-07 13. Béringue V, Le Dur A, Tixador P, Reine F, Lepourry L, Perret-Liaudet A, et al. Prominent and persistent extraneural infection in human PrP transgenic mice infected with variant CJD. PLoS One. 2008;3:e1419. PubMed DOI: 10.1371/journal.pone.0001419 14. Hecker R, Taraboulos A, Scott M, Pan KM, Yang SL, Torchia M, et al. Replication of distinct scrapie prion isolates is region specific in brains of transgenic mice and hamsters. Genes Dev. 1992;6:1213–28. PubMed DOI: 10.1101/gad.6.7.1213 15. Béringue V, Vilotte JL, Laude H. Prion agent diversity and species barrier. Vet Res. 2008;39:47. PubMed DOI: 10.1051/vetres:2008024 Address for correspondence: Vincent Béringue, Institut National de la Recherche Agronomique, UR892, Virologie Immunologie Moléculaires, F-78350 Jouy-en-Josas, France; email: vincent.beringue@jouy.inra.fr Table. Transmission of classical and atypical BSE isolates to transgenic mice expressing human prion protein (Met129)* 1st passage 2nd passage 3rd passage Isolate Origin (identification no.) Total affected† Mean survival time‡ Total affected† Mean survival time‡ Total affected† Mean survival time‡ BSE France (3) 1/6§ 872 6/7 568 ± 65 8/8 523 ± 22 France (3) 2/6 627, 842 Germany¶ 1/4§ 802 6/6 677 ± 54 8/8 555 ± 24 Italy (128204) 0/5 606–775 Belgium 0/4 696–829 L-type Italy (1088) 9/9 607 ± 23 11/11 653 ± 13 NA France (7) 7/7 574 ± 35 0/8# 450 France (10) 8/8 703 ± 19 France (11) 9/9 647 ± 26 H-type France (1) 0/6 376–721 0/7 350–850 NA France (2) 0/6 313–626 0/8 302–755 NA France (5) 0/10 355–838 NA *BSE, bovine spongiform encephalopathy; NA, not available (experiments still ongoing). †Mice with neurologic signs and positive for protease-resistant prion protein by Western blotting. ‡Days ± SE of the mean. For mice with negative results, only the range of survival time is given. §First passage performed on hemizygous mice. Note that the primary transmission of France (3) isolate was performed on both hemizygous and homozygous mice. ¶One passage on transgenic mice expressing bovine prion protein (7). #Ongoing experiment. Page 6 of 8 Figure 1. Protease-resistant prion protein (PrPres) in the brains of human PrP transgenic mice infected with atypical or zoonotic bovine spongiform encephalopathy (BSE) agents. A) Representative Western blot analysis of PrPres extracted (for detailed protocol, see 7) from brain homogenates of mice at terminal stage of disease or at end of lifespan after serial transmission of atypical (L-type and H-type) or classical BSE isolates. The amount of equivalent brain tissue loaded onto the gels was 0.01 mg (BSE; Fr3 isolate, 2nd and 3rd passage), 0.3 mg (L-type and 1st passage of BSE), and 10 mg (PrPres-negative samples). Anti-PrP monoclonal antibody Sha31 was used for PrPres detection. Immunoreactivity was determined by chemiluminescence. B) Ratio of diglycosylated and monoglycosylated PrPres species in the brains of mice after serial transmission of L-type or BSE isolates (data plotted as means ± standard error of the mean). Primary passage of L-type isolates are represented as triangles (orange, It; blue, Fr7; green, Fr10; pink, Fr11) and BSE as squares (light blue, Fr3; red, Ge). Passages are indicated by unfilled symbols of the same color (solid line, second passage; broken line, third passage). The ratio was determined after acquisition of PrPres chemiluminescent signals with a digital imager as previously described (7). Note the distinct glycoform ratio between L-type and BSE groups. It, Italy; Fr, France; Ge, Germany. Page 7 of 8 Figure 2. Representative histoblots in 4 different anteroposterior sections showing the distribution of disease-specific PrPres deposits in the brains of tg650 mice infected with bovine spongiform encephalopathy (BSE) or L-type BSE. Panels A–D show infection with BSE (second passage of France 3). Panels E–H show infection with L-type BSE (first passage of France 7). Panels I–L show infection with L-type BSE (second passage of Italy). Panels M–P show brain sections of an age-matched, mock-infected mouse, euthanized while healthy at 700 days postinfection, for comparison. Note the differing aspect and distribution of PrPres deposits between brain of mice infected with BSE and BSE-L (arrowheads). Assignment of the positive brain regions has been made according to a mouse brain atlas after digital acquisition. Page 8 of 8


Tuesday, August 19, 2008

Atypical BSE (BASE) Transmitted from Asymptomatic Aging Cattle to a Primate


Review on the epidemiology and dynamics of BSE epidemics

Vet. Res. (2008) 39:15 www.vetres.org DOI: 10.1051/vetres:2007053 c INRA, EDP Sciences, 2008 Review article


And last but not least, similarities of PrPres between Htype BSE and human prion diseases like CJD or GSS have been put forward [10], as well as between L-type BSE and CJD [17]. These findings raise questions about the origin and inter species transmission of these prion diseases that were discovered through the BSE active surveillance.


Cases of atypical BSE have only been found in countries having implemented large active surveillance programs. As of 1st September 2007, 36 cases (16 H, 20 L) have been described all over the world in cattle: Belgium (1 L) [23], Canada (1 H)15, Denmark (1 L)16, France (8 H, 6 L)17, Germany (1 H, 1 L) [13], Italy (3 L)18, Japan (1 L) [71], Netherlands (1 H, 2 L)19, Poland (1 H, 6 L)20, Sweden (1 H)21, United Kingdom (1 H)22, and USA (2 H)23. Another H-type case has been found in a 19 year old miniature zebu in a zoological park in Switzerland [56]. It is noteworthy that atypical cases have been found in countries that did not experience classical BSE so far, like Sweden, or in which only few cases of classical BSE have been found, like Canada or the USA.

And last but not least, similarities of PrPres between Htype BSE and human prion diseases like CJD or GSS have been put forward [10], as well as between L-type BSE and CJD [17]. These findings raise questions about the origin and inter species transmission of these prion diseases that were discovered through the BSE active surveillance.

full text 18 pages ;


please see full text ;


***Atypical forms of BSE have emerged which, although rare, appear to be more virulent than the classical BSE that causes vCJD.***

Progress Report from the National Prion Disease Pathology Surveillance Center

An Update from Stephen M. Sergay, MB, BCh & Pierluigi Gambetti, MD

April 3, 2008


Sunday, March 16, 2008

MAD COW DISEASE terminology UK c-BSE (typical), atypical BSE H or L, and or Italian L-BASE


Wednesday, October 08, 2008

Idiopathic Brainstem Neuronal Chromatolysis (IBNC): a novel prion protein related disorder of cattle?


OIE Recognition of the BSE Status of Members RESOLUTION No. XXI (Adopted by the International Committee of the OIE on 27 May 2008)

snip...SEE FULL TEXT with facts and sources @ ;



Friday, April 25, 2008

Substances Prohibited From Use in Animal Food or Feed [Docket No. 2002N-0273] (Formerly Docket No. 02N-0273) RIN 0910-AF46


HUMAN and ANIMAL TSE Classifications i.e. mad cow disease and the UKBSEnvCJD only theory JUNE 2008


Tissue infectivity and strain typing of the many variants of the human and animal TSEs are paramount in all variants of all TSE. There must be a proper classification that will differentiate between all these human TSE in order to do this. With the CDI and other more sensitive testing coming about, I only hope that my proposal will some day be taken seriously. ...




Oct 23, 2008 at 9:00 AM



***The pathology features of Nor98 in the cerebellum of the affected sheep showed similarities with those of sporadic Creutzfeldt-Jakob disease in humans.


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.

Edited by Stanley B. Prusiner, University of California, San Francisco, CA, and approved September 12, 2005 (received for review March 21, 2005)




NOR-98 ATYPICAL SCRAPIE 5 cases documented in USA in 5 different states USA 007





Sunday, September 07, 2008

CWD LIVE TEST, and the political aspects or fallout of live testing for BSE in cattle in the USA





Saturday, December 01, 2007

Phenotypic Similarity of Transmissible Mink Encephalopathy in Cattle and L-type Bovine Spongiform Encephalopathy in a Mouse

Model Volume 13, Number 12–December 2007 Research

Phenotypic Similarity of Transmissible Mink Encephalopathy in Cattle andL-type Bovine Spongiform Encephalopathy in a Mouse Model

Thierry Baron,* Anna Bencsik,* Anne-Gaëlle Biacabe,* Eric Morignat,* andRichard A. Bessen†*Agence Française de Sécurité Sanitaire des Aliments–Lyon, Lyon, France; and†Montana State University, Bozeman, Montana, USA


Transmissible mink encepholapathy (TME) is a foodborne transmissiblespongiform encephalopathy (TSE) of ranch-raised mink; infection with aruminant TSE has been proposed as the cause, but the precise origin of TMEis unknown. To compare the phenotypes of each TSE, bovine-passaged TMEisolate and 3 distinct natural bovine spongiform encephalopathy (BSE) agents(typical BSE, H-type BSE, and L-type BSE) were inoculated into an ovinetransgenic mouse line (TgOvPrP4). Transgenic mice were susceptible toinfection with bovine-passaged TME, typical BSE, and L-type BSE but not toH-type BSE. Based on survival periods, brain lesions profiles,disease-associated prion protein brain distribution, and biochemicalproperties of protease-resistant prion protein, typical BSE had a distintphenotype in ovine transgenic mice compared to L-type BSE and bovine TME.The similar phenotypic properties of L-type BSE and bovine TME in TgOvPrP4mice suggest that L-type BSE is a much more likely candidate for the originof TME than is typical BSE.



These studies provide experimental evidence that the Stetsonville TME agentis distinct from typical BSE but has phenotypic similarities to L-type BSEin TgOvPrP4 mice. Our conclusion is that L-type BSE is a more likelycandidate for a bovine source of TME infection than typical BSE. In thescenario that a ruminant TSE is the source for TME infection in mink, thiswould be a second example of transmission of a TSE from ruminants tonon-ruminants under natural conditions or farming practices in addition totransmission of typical BSE to humans, domestic cats, and exotic zoo animals(37). The potential importance of this finding is relevant to L-type BSE,which based on experimental transmission into humanized PrP transgenic miceand macaques, suggests that L-type BSE is more pathogenic for humans thantypical BSE (24,38).


Transmissible Mink Encephalopathy TME Subject: In Confidence - Perceptions of unconventional slow virus diseasesof animals in the USA - APRIL-MAY 1989 - G A H Wells

Gerald Wells: Report of the Visit to USA, April-May 1989


The general opinion of those present was that BSE, as anovert disease phenomenon, _could exist in the USA, but if it did,it was very rare. The need for improved and specific surveillancemethods to detect it as recognised...


It is clear that USDA have little information and _no_ regulatoryresponsibility for rendering plants in the US...


3. Prof. A. Robertson gave a brief account of BSE. The US approachwas to accord it a _very low profile indeed_. Dr. A Thiermann showedthe picture in the ''Independent'' with cattle being incinerated and thoughtthis was a fanatical incident to be _avoided_ in the US _at all costs_...

snip...please read this old full text document !


To be published in the Proceedings of theFourth International Scientific Congress inFur Animal Production. Toronto, Canada,August 21-28, 1988

Evidence That Transmissible Mink EncephalopathyResults from Feeding Infected Cattle

R.F. Marsh* and G.R. Hartsough

•Department of Veterinary Science, University of Wisconsin-Madison, Madison,Wisconsin 53706; and ^Emba/Creat Lakes Ranch Service, Thiensville, Wisconsin53092

ABSTRACT Epidemiologic investigation of a new incidence oftransmissible mink encephalopathy (TME) in Stetsonville, Wisconsinsuggests that the disease may have resulted from feeding infectedcattle to mink. This observation is supported by the transmission ofa TME-like disease to experimentally inoculated cattle, and by therecent report of a new bovine spongiform encephalopathy inEngland.


Transmissible mink encephalopathy (TME) was first reported in 1965 by Hartsoughand Burger who demonstrated that the disease was transmissible with a long incubationperiod, and that affected mink had a spongiform encephalopathy similar to that found inscrapie-affecied sheep (Hartsough and Burger, 1965; Burger and Hartsough, 1965).Because of the similarity between TME and scrapie, and the subsequent finding that thetwo transmissible agents were indistinguishable (Marsh and Hanson, 1969), it wasconcluded that TME most likely resulted from feeding mink scrapie-infecied sheep.The experimental transmission of sheep scrapie to mink (Hanson et al., 1971)confirmed the close association of TME and scrapie, but at the same time providedevidence that they may be different. Epidemiologic studies on previous incidences ofTME indicated that the incubation periods in field cases were between six months andone year in length (Harxsough and Burger, 1965). Experimentally, scrapie could not betransmitted to mink in less than one year.To investigate the possibility that TME may be caused by a (particular strain ofscrapie which might be highly pathogenic for mink, 21 different strains of the scrapieagent, including their sheep or goat sources, were inoculated into a total of 61 mink.Only one mink developed a progressive neurologic disease after an incubation period of22 mon..s (Marsh and Hanson, 1979). These results indicated that TME was either causedby a strain of sheep scrapie not yet tested, or was due to exposure to a scrapie-like agentfrom an unidentified source.


A New Incidence of TME. In April of 1985, a mink rancher in Stetsonville, Wisconsinreported that many of his mink were "acting funny", and some had died. At this time, wevisited the farm and found that approximately 10% of all adult mink were showingtypical signs of TME: insidious onset characterized by subtle behavioral changes, loss ofnormal habits of cleanliness, deposition of droppings throughout the pen rather than in asingle area, hyperexcitability, difficulty in chewing and swallowing, and tails arched overtheir _backs like squirrels. These signs were followed by progressive deterioration ofneurologic function beginning with locomoior incoordination, long periods of somnolencein which the affected mink would stand motionless with its head in the corner of thecage, complete debilitation, and death. Over the next 8-10 weeks, approximately 40% ofall the adult mink on the farm died from TME.Since previous incidences of TME were associated with common or shared feedingpractices, we obtained a careful history of feed ingredients used over the past 12-18months. The rancher was a "dead stock" feeder using mostly (>95%) downer or dead dairycattle and a few horses. Sheep had never been fed.

Experimental Transmission. The clinical diagnosis of TME was confirmed byhistopaihologic examination and by experimental transmission to mink after incubationperiods of four months. To investigate the possible involvement of cattle in this diseasecycle, two six-week old castrated Holstein bull calves were inoculated intracerebrallywith a brain suspension from affected mink. Each developed a fatal spongiformencephalopathy after incubation periods of 18 and 19 months.


These findings suggest that TME may result from feeding mink infected cattle andwe have alerted bovine practitioners that there may exist an as yet unrecognizedscrapie-like disease of cattle in the United States (Marsh and Hartsough, 1986). A newbovine spongiform encephalopathy has recently been reported in England (Wells et al.,1987), and investigators are presently studying its transmissibility and possiblerelationship to scrapie. Because this new bovine disease in England is characterized bybehavioral changes, hyperexcitability, and agressiveness, it is very likely it would beconfused with rabies in the United Stales and not be diagnosed. Presently, brains fromcattle in the United States which are suspected of rabies infection are only tested withanti-rabies virus antibody and are not examined histopathologically for lesions ofspongiform encephalopathy.We are presently pursuing additional studies to further examine the possibleinvolvement of cattle in the epidemiology of TME. One of these is the backpassage ofour experimental bovine encephalopathy to mink. Because (here are as yet no agent-specific proteins or nucleic acids identified for these transmissible neuropathogens, onemeans of distinguishing them is by animal passage and selection of the biotype whichgrows best in a particular host. This procedure has been used to separate hamster-adapted and mink-udapted TME agents (Marsh and Hanson, 1979). The intracerebralbackpassage of the experimental bovine agent resulted in incubations of only four monthsindicating no de-adaptation of the Stetsonville agent for mink after bovine passage.Mink fed infected bovine brain remain normal after six months. It will be essential todemonstrate oral transmission fiom bovine to mink it this proposed epidemiologicassociation is to be confirmed.


These studies were supported by the College of Agricultural and Life Sciences,University of Wisconsin-Madison and by a grant (85-CRCR-1-1812) from the UnitedStates Department of Agriculture. The authors also wish to acknowledge the help andencouragement of Robert Hanson who died during the course of these investigations.


Burger, D. and Hartsough, G.R. 1965. Encephalopathy of mink. II. Experimental andnatural transmission. J. Infec. Dis. 115:393-399.Hanson, R.P., Eckroade, R.3., Marsh, R.F., ZuRhein, C.M., Kanitz, C.L. and Gustatson,D.P. 1971. Susceptibility of mink to sheep scrapie. Science 172:859-861.Hansough, G.R. and Burger, D. 1965. Encephalopathy of mink. I. Epizoociologic andclinical observations. 3. Infec. Dis. 115:387-392.Marsh, R.F. and Hanson, R.P. 1969. Physical and chemical properties of thetransmissible mink encephalopathy agent. 3. ViroL 3:176-180.Marsh, R.F. and Hanson, R.P. 1979. On the origin of transmissible minkencephalopathy. In Hadlow, W.J. and Prusiner, S.P. (eds.) Slow transmissiblediseases of the nervous system. Vol. 1, Academic Press, New York, pp 451-460.Marsh, R.F. and Hartsough, G.R. 1986. Is there a scrapie-like disease incattle? Proceedings of the Seventh Annual Western Conference for Food AnimalVeterinary Medicine. University of Arizona, pp 20.Wells, G.A.H., Scott, A.C., Johnson, C.T., Cunning, R.F., Hancock, R.D.,Jeffrey, M., Dawson, M. and Bradley, R. 1987. A novel progressive spongiformencephalopathy in cattle. Vet. Rec. 121:419-420.



In Confidence - Perceptions of unconventional slow virus diseasesof animals in the USA - APRIL-MAY 1989 - G A H Wells


Wednesday, August 20, 2008

Bovine Spongiform Encephalopathy Mad Cow Disease typical and atypical strains, was there a cover-up ?


Subject: TME hyper/drowsy, INTER-SPECIES TRANSMISSION CWD and strainpropertiesDate: October 22, 2007 at 12:48 pm PST

Completely Edited Version


TME hyper/drowsy, INTER-SPECIES TRANSMISSION CWD and strain properties

page 19 of 62. ...tss

Dr. Detwiler: How would you explain that biochemically?

snip...see full text ;


A New Prionopathy OR more of the same old BSe and sporadic CJD


Communicated by: Terry S. Singeltary Sr.

[In submitting these data, Terry S. Singeltary Sr. draws attention to the steady increase in the "type unknown" category, which, according to their definition, comprises cases in which vCJD could be excluded. The total of 26 cases for the current year (2007) is disturbing, possibly symptomatic of the circulation of novel agents. Characterization of these agents should be given a high priority. - Mod.CP]



There is a growing number of human CJD cases, and they were presented last week in San Francisco by Luigi Gambatti(?) from his CJD surveillance collection.

He estimates that it may be up to 14 or 15 persons which display selectively SPRPSC and practically no detected RPRPSC proteins.



sporadic Fatal Familial Insomnia



MARCH 26, 2003

RE-Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob

disease in the United States

Email Terry S. Singeltary:


I lost my mother to hvCJD (Heidenhain Variant CJD). I would like to comment on the CDC's attempts to monitor the occurrence of emerging forms of CJD. Asante, Collinge et al [1] have reported that BSE transmission to the 129-methionine genotype can lead to an alternate phenotype that is indistinguishable from type 2 PrPSc, the commonest sporadic CJD. However, CJD and all human TSEs are not reportable nationally. CJD and all human TSEs must be made reportable in every state and internationally. I hope that the CDC does not continue to expect us to still believe that the 85%+ of all CJD cases which are sporadic are all spontaneous, without route/source. We have many TSEs in the USA in both animal and man. CWD in deer/elk is spreading rapidly and CWD does transmit to mink, ferret, cattle, and squirrel monkey by intracerebral inoculation. With the known incubation periods in other TSEs, oral transmission studies of CWD may take much longer. Every victim/family of CJD/TSEs should be asked about route and source of this agent. To prolong this will only spread the agent and needlessly expose others. In light of the findings of Asante and Collinge et al, there should be drastic measures to safeguard the medical and surgical arena from sporadic CJDs and all human TSEs. I only ponder how many sporadic CJDs in the USA are type 2 PrPSc?



Hardcover, 304 pages plus photos and illustrations. ISBN 0-387-95508-9

June 2003

BY Philip Yam


Answering critics like Terry Singeltary, who feels that the U.S. under- counts CJD, Schonberger conceded that the current surveillance system has errors but stated that most of the errors will be confined to the older population.


Diagnosis and Reporting of Creutzfeldt-Jakob Disease Singeltary, Sr et al. JAMA.2001; 285: 733-734. Vol. 285 No. 6, February 14, 2001 JAMA

Diagnosis and Reporting of Creutzfeldt-Jakob Disease

To the Editor: In their Research Letter, Dr Gibbons and colleagues1 reported that the annual US death rate due to Creutzfeldt-Jakob disease (CJD) has been stable since 1985. These estimates, however, are based only on reported cases, and do not include misdiagnosed or preclinical cases. It seems to me that misdiagnosis alone would drastically change these figures. An unknown number of persons with a diagnosis of Alzheimer disease in fact may have CJD, although only a small number of these patients receive the postmortem examination necessary to make this diagnosis. Furthermore, only a few states have made CJD reportable. Human and animal transmissible spongiform encephalopathies should be reportable nationwide and internationally.

Terry S. Singeltary, Sr Bacliff, Tex

1. Gibbons RV, Holman RC, Belay ED, Schonberger LB. Creutzfeldt-Jakob disease in the United States: 1979-1998. JAMA. 2000;284:2322-2323. FREE FULL TEXT



2 January 2000 British Medical Journal U.S. Scientist should be concerned with a CJD epidemic in the U.S., as well


15 November 1999 British Medical Journal vCJD in the USA * BSE in U.S.


Creutzfeldt Jakob Disease




Sunday, April 20, 2008 Progress Report from the National Prion Disease Pathology Surveillance Center April 3, 2008

Atypical forms of BSE have emerged which, although rare, appear to be more virulent than the classical BSE that causes vCJD.

see full text ;


CJD TEXAS (cjd clusters)




The statistical incidence of CJD cases in the United States has been revised to reflect that there is one case per 9000 in adults age 55 and older. Eighty-five percent of the cases are sporadic, meaning there is no known cause at present.


Terry S. Singeltary Sr.
P.O. Box 42
Bacliff, Texas USA 77518

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