Tuesday, November 04, 2014

The pathological and molecular but not clinical phenotypes are maintained after second passage of experimental atypical bovine spongiform encephalopathy in cattle

The pathological and molecular but not clinical phenotypes are maintained after second passage of experimental atypical bovine spongiform encephalopathy in cattle
Timm Konold, Laura J Phelan, [...], and Marion M Simmons

Additional article information

Abstract Background Atypical bovine spongiform encephalopathies (BSEs), classified as H-type and L-type BSE based on the Western immunoblot profiles, are naturally occurring diseases in cattle, which are phenotypically different to classical BSE. Transmission studies in cattle using the intracerebral route resulted in disease where the phenotypes were maintained irrespective of BSE type but clinically affected cattle with a shorter survival time displayed a nervous form whereas cattle with a longer survival time displayed a dull form. A second transmission study is reported here where four cattle were intracerebrally inoculated with brain tissue from experimentally infected cattle presenting with either the nervous or dull form of H- or L-type BSE to determine whether the phenotype is maintained.

Results The four inoculated cattle were culled at 16.5-19.5 months post inoculation after presenting with difficulty getting up, a positive scratch response (all) and dullness (three cattle), which was not observed in two non-inoculated control cattle, each housed with either group of inoculated cattle. Only the inoculated cattle had detectable prion protein in the brain based on immunohistochemical examination, and the Western immunoblot profile was consistent with the H-type or L-type BSE of the respective donor cattle.


Conclusions Second passage of H-type and L-type BSE in cattle produced a TSE where the majority of cattle displayed the dull form regardless of clinical disease form of the donor cattle. The pathological and molecular phenotypes of H- and L-type BSE were maintained.


Electronic supplementary material The online version of this article (doi:10.1186/s12917-014-0243-2) contains supplementary material, which is available to authorized users.


Conclusions Second passage of H-type and L-type BSE in cattle by intracerebral inoculation produced a TSE where the majority displayed the dull disease form regardless of disease form of the donor cattle. The pathological and molecular phenotypes of H- and L-type BSE were maintained.



 ISSN 1999-4915


www.mdpi.com/journal/viruses Article


Molecular Modeling of Prion Transmission to Humans


Etienne Levavasseur 1, Nicolas Privat 1, Juan-Carlos Espinosa Martin 2, Steve Simoneau 3, Thierry Baron 4, Benoit Flan 3, Juan-Maria Torres 2 and Stéphane Haïk 1,5,6,* 1 Inserm U 1127, CNRS UMR 7225, Sorbonne Universités, UPMC Univ. Paris 06 UMR S 1127, Institut du Cerveau et de la Moelle épinière, ICM, 75013 Paris, France; E-Mails: etienne.levavasseur@inserm.fr (E.L.); nicolas.privat@inserm.fr (N.P.) 2 Centro de Investigacion en Sanidad Animal, Carretera de Algete a El Casar, 28130 Madrid, Spain; E-Mails: espinosa.juan@inia.es (J.-C.E.M.); jmtorres@inia.es (J.-M.T.) 3 LFB Biomédicaments, 91958 Les Ulis, France; E-Mails: simoneaus@lfb.fr (S.S.); flan@lfb.fr (B.F.) 4 Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail (ANSES), Unité Maladies neurodégénératives, 69394 Lyon, France; E-Mail: thierry.baron@anses.fr 5 AP-HP, Hôpital de la Pitié-Salpêtrière, Cellule nationale de référence des MCJ, F-75013 Paris, France 6 AP-HP, Hôpital de la Pitié-Salpêtrière, Neuropathologie, 75013 Paris, France * Author to whom correspondence should be addressed; E-Mail: stephane.haik@courriel.upmc.fr; Tel.: +33-142-162-628; Fax: +33-142-161-899.


External Editor: Judd Aiken and Debbie McKenzie


Received: 24 July 2014; in revised form: 26 September 2014/ Accepted: 30 September 2014/ Published: 02 October 2014


Abstract: Using different prion strains, such as the variant Creutzfeldt-Jakob disease agent and the atypical bovine spongiform encephalopathy agents, and using transgenic mice expressing human or bovine prion protein, we assessed the reliability of protein misfolding cyclic amplification (PMCA) to model interspecies and genetic barriers to prion transmission. We compared our PMCA results with in vivo transmission data characterized by attack rates, i.e., the percentage of inoculated mice that developed the disease. Using 19 seed/substrate combinations, we observed that a significant PMCA amplification was only obtained when the mouse line used as substrate is susceptible to the corresponding strain. Our results suggest that PMCA provides a useful tool to study genetic barriers to transmission and to study the zoonotic potential of emerging prion strains.


1. Introduction


Prion diseases are fatal transmissible disorders affecting humans and animals. They are characterized by brain vacuolization, neuronal loss and accumulation of PrPsc, an abnormal isoform of the host-encoded cellular prion protein (PrPc). PrPsc has been proposed as the infectious agent, capable of converting PrPc into PrPsc in an autocatalytical manner [1]. In humans, prion diseases result from contamination, genetic inheritance or sporadic event. The host susceptibility is influenced by the prion protein-encoding gene PRNP. For example, the variant of Creutzfeldt-Jakob disease (vCJD), which has been associated to the classical bovine spongiform encephalopathy (C-BSE) epidemics in cattle through contaminated meat product consumption [2], has occurred so far only in individuals homozygous for methionine at codon 129 of PRNP [3]. However, this finding has been under debate [4,5]. Studies have investigated interspecies and genotypic barriers [6,7] using an in vitro PrPsc amplification system named protein misfolding cyclic amplification (PMCA) [8]. This method allows, in PCR tubes, the amplification of minute amounts of PrPsc in infected tissues (seed) in the presence of normal brain homogenate in excess (substrate), after cycles of incubation and sonication. Then, the final product of the reaction can be detected after proteinase K digestion by Western blot. While PMCA allows the amplification of PrPsc, it has also been demonstrated that infectivity was increased during the reaction [9], and that prion strain properties were maintained throughout the reaction [10–12]. Brains from humans or transgenic mice expressing a human PrP with methionine at codon 129 of PRNP provided the best substrates to amplify vCJD and BSE PrPsc [6,7], suggesting that PMCA may reproduce faithfully the genotypic transmission barrier. It was thus proposed as a means to evaluate the zoonotic risk associated with emerging prion strains (Nor98 in sheep, L-type BSE in cattle). Indeed, while classical BSE strain has been recognized to be at the origin of vCJD in humans, L-BSE is considered to be a sporadic form of prion disease in cattle, differing in many aspects (epidemiology, neuropathology, biochemical features) from the C-BSE strain.


***Moreover, L-BSE has been transmitted more easily to transgenic mice overexpressing a human PrP [13,14] or to primates [15,16] than C-BSE.


***It has been suggested that some sporadic CJD subtypes in humans may result from an exposure to the L-BSE agent. Lending support to this hypothesis, pathological and biochemical similarities have been observed between L-BSE and an sCJD subtype (MV genotype at codon 129 of PRNP) [17], and between L-BSE infected non-human primate and another sCJD subtype (MM genotype) [15].





***P.170: Potential detection of oral transmission of H type atypical BSE in cattle using in vitro conversion


Sandor Dudas, John G Gray, Renee Clark, and Stefanie Czub Canadian Food Inspection Agency; Lethbridge, AB Canada


Keywords: Atypical BSE, oral transmission, RT-QuIC


The detection of bovine spongiform encephalopathy (BSE) has had a significant negative impact on the cattle industry worldwide. In response, governments took actions to prevent transmission and additional threats to animal health and food safety. While these measures seem to be effective for controlling classical BSE, the more recently discovered atypical BSE has presented a new challenge. To generate data for risk assessment and control measures, we have challenged cattle orally with atypical BSE to determine transmissibility and mis-folded prion (PrPSc) tissue distribution. Upon presentation of clinical symptoms, animals were euthanized and tested for characteristic histopathological changes as well as PrPSc deposition.


The H-type challenged animal displayed vacuolation exclusively in rostral brain areas but the L-type challenged animal showed no evidence thereof. To our surprise, neither of the animals euthanized, which were displaying clinical signs indicative of BSE, showed conclusive mis-folded prion accumulation in the brain or gut using standard molecular or immunohistochemical assays. To confirm presence or absence of prion infectivity, we employed an optimized real-time quaking induced conversion (RT-QuIC) assay developed at the Rocky Mountain Laboratory, Hamilton, USA.


Detection of PrPSc was unsuccessful for brain samples tests from the orally inoculated L type animal using the RT-QuIC. It is possible that these negative results were related to the tissue sampling locations or that type specific optimization is needed to detect PrPSc in this animal. We were however able to consistently detect the presence of mis-folded prions in the brain of the H-type inoculated animal. Considering the negative and inconclusive results with other PrPSc detection methods, positive results using the optimized RT-QuIC suggests the method is extremely sensitive for H-type BSE detection. This may be evidence of the first successful oral transmission of H type atypical BSE in cattle and additional investigation of samples from these animals are ongoing.


P.169: PrPSc distribution in brain areas of a natural German H-type BSE case


Anne Balkema-Buschmann, Grit Priemer, Markus Keller, and Martin H Groschup Friedrich Loeffler Institut, Institute for Novel and Emerging Infectious Diseases; Greifswald, Insel Riems, Germany


Keywords: BSE H-type, brain, muscle


Ten years after the initial description of atypical BSE cases of the H-type and L-type, the distribution of PrPSc in different brain areas and peripheral tissues of natural cases of these BSE forms is still not fully understood. Intracerebral challenge experiments have been performed with both atypical BSE forms in cattle, and the distribution of the abnormal prion protein and infectivity has been analysed in a variety of tissues, confirming the general restriction to the central nervous system as it was already generally acknowledged for classical BSE, but showing a slightly earlier and stronger involvement of the peripheral nervous system and the skeletal muscle. www.landesbioscience.com Prion 105


However, data from cattle orally challenged with atypical BSE, which might mimic the natural situation, are not yet available. Unfortunately, for most natural cases of atypical BSE, only the obex region is available for further analysis. The PrPSc distribution in the brains of natural L-type BSE cases in Italy has been described in some detail, but comparably few such data are yet available for natural H-type cases. Here we describe the analysis of different brain areas and muscle samples of a natural H-type BSE case diagnosed in Germany in 2014, and compare these data with those obtained from the respective samples collected from cattle challenged intracerebrally with H-type BSE.


P.159: Transgenic mice overexpressing rabbit prion protein are susceptible to BSE, BASE and scrapie prion strains but resistant to CWD and atypical scrapie


Natalia Fernández-Borges,1 Enric Vidal,2 Belén Pintado,4 Hasier Eraña,1 Montserrat Ordóñez,3 Mercedes Márquez,5 Francesca Chianini,6 Dolors Fondevila,5 Manuel A Sánchez-Martín,7 Olivier Andréoletti,8 Mark P Dagleish,6 Martí Pumarola,5 and Joaquín Castilla1,3 1CIC bioGUNE; Parque tecnológico de Bizkaia; Derio; Bizkaia, Spain; 2Centre de Recerca en Sanitat Animal (CReSA); UAB-IR TA, Campus de la Universitat Autònoma de Barcelona; Bellaterra; Barcelona, Catalonia, Spain; 3IKERBASQUE; Basque Foundation for Science; Bilbao, Bizkaia, Spain; 4Centro Nacional de Biotecnología (CNB), Campus de Cantoblanco; Cantoblanco; Madrid, Spain; 5Department of Animal Medicine and Surgery; Veterinary faculty; Universitat Autònoma de Barcelona (UAB); Bellaterra (Cerdanyola del Vallès); Barcelona, Catalonia, Spain; 6Moredun Research Institute; Bush Loan, Penicuik, Scotland, UK; 7Unidad de Generación de OMGs. S.E.A. Department of Medicine; University of Salamanca; Salamanca, Spain; 8Ecole Nationale du Veterinaire; Service de Pathologie du Bétail; Toulouse, France


Interspecies transmission of prions is a well established phenomenon, both experimentally and in field conditions. Upon passage through new hosts prion strains have proven their capacity to change their properties. It is, in fact, a source of strain diversity which needs to be considered when assessing the potential risks associated with consumption of prion contaminated protein sources.


Rabbits were considered for decades a prion resistant species until proven recently otherwise. To determine the extent of rabbit susceptibility to prions and to assess their effects on the passage of different prion strains through this species, a transgenic mouse model overexpressing rabbit PrPC was developed (TgRab). Intracerebral challenges with prion strains originating from a variety of species including field isolates (SSBP1 scrapie, Nor98-like scrapie, BSE, BASE and CWD), experimental murine strains (ME7 and RML), experimentally obtained strains (sheepBSE) and strains obtained by in vitro crossing of the species barrier using saPMCA (BSE-RabPrPres, SSBP1-RabPrPres and CWD-RabPrPres) have been performed.


Interestingly, on first passage, TgRab were susceptible to the majority of prions tested with the exception of SSBP1 scrapie, CWD and Nor98 scrapie. Furthermore TgRab were capable of propagating strain-specific features such as differences in incubation periods, brain lesion and PrPd deposition profiles and PK resistant western blotting band patterns. Our results confirm previous studies shattering the myth that rabbits are resistant to prion infection and this should be taken into account when choosing protein sources to feed rabbits.


P.168: Evolution of the biological properties of L-BSE after passage in sheep with susceptible and resistant PrP genotypes


Michele A Di Bari, Umberto Agrimi, Claudia D’Agostino, Geraldina Riccardi, Stefano Marcon, Elena Esposito, Paolo Frassanito, Flavio Torriani, Shimon Simson, and Romolo Nonno Istituto Superiore di Sanità (ISS) Department of Veterinary Public Health and Food Safety; Rome, Italy


Background. Cattle L-BSE was efficiently transmitted to sheep with susceptible (QQ171) and resistant (QR171) PrP genotypes. 1 Notably, the PrPSc signature of L-BSE was preserved in QQ171 sheep but not in QR171 sheep.2 Notwithstanding, bioassay in transgenic mice expressing bovine or ovine (ARQ) PrPC showed that L-BSE strain was preserved in both, QQ171 and QR171 sheep-passaged L-BSE.3


Here we studied the biological properties of sheep-passaged L-BSE by bioassay in bank voles and transgenic mice expressing the ovine VRQ PrP (tg338), both characterized by a comparatively low susceptibility to cattle L-BSE.


Material and Methods. Voles and tg338 mice were intracerebrally inoculated with cattle L-BSE and sheep-passaged (QQ171 and QR171) L-BSE isolates. Survival time, lesion profiles, Pet-blot and WB analysis were used for strain typing. Results. Cattle L-BSE transmitted quite inefficiently to tg338 mice, with survival time >400 days post-infection (d.p.i.), while sheep-passaged inocula were much more efficient and all gave terminal disease by ~140 d.p.i. However, after sub-passage all inocula converged to a survival time of ~145 d.p.i.. and showed overlapping pathological phenotypes.


In voles, cattle L-BSE transmitted with very long survival times (~800 d.p.i.) and was accompanied by an upward shift of the PrPSc type. Again, all sheep-passaged L-BSE isolates transmitted much more efficiently, with similar survival times of ~360 d.p.i.. Upon second passage, three different strains were isolated in vole, characterized by distinct pathological phenotypes. This divergence is epitomized by the different survival times of vole-adapted L-BSE strains, which were ~400 d.p.i. for cattle L-BSE, ~130 d.p.i. for QQ171-passaged L-BSE and ~225 d.p.i. for QR171-passaged L-BSE.


Conclusions. These findings, along with previously published data,3 show that the original L-BSE strain was recovered after passage in sheep when bioassay was performed in animal models expressing bovine or ovine PrPC. In contrast, strain changes were observed in both, QQ171- and QR171-passaged L-BSE by bioassay in vole, a species with divergent PrP sequence compared to ruminants. Importantly, QQ171- and QR171-passaged L-BSE were characterised by different PrPSc types and, accordingly, showed different biological properties when transmitted to voles, but not when transmitted to other animal models.


Overall, our work support the hypothesis that prion isolates are likely composed of multiple prion components, emphasizes the role of host PrP polymorphisms on strain selection and mutation, and highlights the risk for new potentially zoonotic strains that could emerge from prion evolution in animal reservoirs.


P.172: BSE exposure risk from bovine intestine and mesentery


Fulvio Barizzone,1 Herbert Budka,2 Christine Fast,3 John N Griffin,4 Giuseppe Ru,5 Pietro Stella1 and Olivier Andréoletti6 1European Food Safety Authority; Parma, Italy; 2Institute of Neuropathology; University Hospital Zurich; Zurich, Switzerland; 3Friedrich-Loeffler-Institut; Institute of Novel and Emerging Infectious Diseases; Isle of Riems, Germany; 4Department of Agriculture, Food and the Marine; Backweston, Celbridge, Co. Kildare, Ireland; 5Istituto Zooprofilattico Sperimentale del Piemonte; Liguria e Valle d’Aosta; Biostatistics Epidemiology and Analysis of Risk (BEAR) unit; Turin, Italy; 6UMR Interactions Hôtes Agents Pathogènes; Ecole Nationale Vétérinaire INR A; ENVT; Toulouse, France


Keywords: Bovine Spongiform Encephalopathy (BSE), cattle, intestine, mesentery, specified risk material (SRM), quantitative risk assessment (QRA)


Bovine intestines and mesenteries in the European Union (EU) are considered among the tissues potentially containing the highest level of BSE infectivity and have to be removed from the food and feed chain. A quantitative assessment of the BSE infectious load potentially entering the food and feed chain yearly in the European Union (EU) was developed. The evolution of the BSE infectious titre and of the weight of the structures accumulating infectivity was considered. The number of BSE infected cattle entering undetected in the food and feed chain yearly was estimated. A model (TSEi) was developed to estimates the evolution of the BSE infectious load in animals and the total yearly infectious load that could enter the food and feed chain. In a BSE infected bovine, the distribution of infectivity in intestines and mesentery varies with the age. Up to 36 months of age the infectivity is mainly associated (on average more than 90%) with the last 4 metres of small intestine and the caecum, over 36 and under 60 months of age, there is an inter-individual variability, from 60 months of age the infectivity is mainly associated (on average more than 90%) with the mesenteric nerves and the celiac and mesenteric ganglion complex. The total amount of infectivity peaks, about 15 BoID50, in animals younger than 18 months, it declines to 8-9 BoID50 (24–48 months of age) and it drops to 0.7 BoID50 in animals older than 60 months. The ileocaecal plate is the most infectious part of the intestine and it can be used to estimate the potential maximum level of exposure for an individual consumer.


In the EU, between 2007 and 2012, the yearly amount of BSE infectivity associated with intestine and mesentery from animals entering the food and feed chain was reduced by a factor of 10 (from about 23,000 to about 2,000 BoID50).


However, the maximum level of exposure to the BSE agent from intestine remained stable (on average about 1.5-1.6 BoID50 per meter).


In case of re-emergence of BSE in the EU there would be an increase of the potential maximum level of exposure to BSE from intestine. According to the TSEi model the removal of the last four metres of the small intestine and of the caecum from the food and feed chain would result in a major reduction of the BSE exposure risk associated with intestine and mesentery in cattle.


P.131: Transmission of sheep-bovine spongiform encephalopathy in pigs


Carlos Hedman,1 Belén Marín,1 Fabian Corbière,3 Hicham Filali,1 Francisco Vázquez, José Luis Pitarch,1 William Jirón,1 Rodrigo S Hernandez,1 Bernardino Moreno,1 Martí Pumarola,2 Olivier Andréoletti,3 Juan José Badiola,1 and Rosa Bolea1 1University of Zaragoza; Zaragoza, Spain; 2University of Barcelona; Barcelona, Spain; 3Institut National de la Recherche (INR A); Toulouse, France


Introduction. The transmissible spongiform encephalopathies (TSE) don´t occur in swine in natural conditions. However, the bovine spongiform encephalopathy (BSE) agent, inoculated by 3 simultaneous routes in pigs, is able to reproduce a neurological disease in these animals. On the other hand, the BSE agent after passage in sheep under experimental conditions (sheep- BSE) exhibits altered pathobiologic properties. This new agent is able to cross the cattle-pig transmission barrier more efficiently than BSE. The potential propagation of TSE in animals from the human food chain, including pigs, needs to be assessed regarding the risk for human infection by animals other than TSE-infected ruminants. The aim of this work was to determine the susceptibility of pigs to the Sheep-BSE agent and describe the pathological findings and PrPSc deposition in different tissues.


Material and Methods. Seven minipigs were challenged intracerebrally with sheep-BSE agent. Clinical observation and postmortem histopathology, immunohistochemistry (antibody 2G11) and Western blotting were performed on central nervous system (CNS), peripheral nervous system (PNS) and other tissues.


Results. One pig was culled in an early incubation stage, and remaining six were culled at the presence of clinical sings. Pigs developed a clinical disease with locomotor disorders in an average time of 23 months post inoculation, showing clinical findings in most of them earlier than those described in the BSE in pigs experimental infection. TSE wasn´t confirmed in the preclinical pig. In clinical pigs, the entire cerebral cortex showed severe neuropil vacuolation, extensive and severe vacuolar changes affecting the thalamus, hippocampus and cerebellum. PrPSc was found in CNS of all clinical pigs (6/6). Intracellular (intraneuronal and intraglial) and neuropil-associated PrPSc deposition was consistently observed in the brainstem, thalamus, and deeper layers of the cerebral cortex. Also, PrPSc was observed in PNS, mainly in the myenteric plexus and also in nerves belonging to the skeleton muscle. Moreover, the glycosylation profile showed a 3 band pattern with a predominant monoglycosylated band in positive pig samples.


This features concern on the potential risk of utilization of meat and bound meal of small ruminants in feeding pigs.


P.177: Elements modulating the prion species barrier and its passage consequences


Juan-Carlos Espinosa,1 Patricia Aguilar-Calvo,1 Ana Villa-Diaz,1 Olivier Andréoletti,2 and Juan María Torres1 1Centro de Investigación en Sanidad Animal (CISA-INI A); Valdeolmos, Madrid, Spain; 2UMR INR A-ENVT 1225; Interactions Hôte Agent Pathogène; École Nationale Vétérinaire de Toulouse; Toulouse, France


The phenotypic features of Transmissible Spongiform Encephalopathy (TSE) strains may be modified during passage across a species barrier. In this study we investigated the biochemical and biological characteristics of Bovine Spongiform Encephalopathy (BSE) infectious agent after transmission in both natural host species (cattle, sheep, pigs, and mice) and in transgenic mice overexpressing the corresponding cellular prion protein (PrPC) in comparison with other non-BSE related prions from the same species. After these passages, most characteristics of the BSE agent remained unchanged. BSE-derived agents only showed slight modifications in the biochemical properties of the accumulated PrPSc, which were demonstrated to be reversible upon re-inoculation into transgenic mice expressing bovine-PrPC. Transmission experiments in transgenic mice expressing bovine, porcine or human-PrP revealed that all BSE-derived agents were transmitted with no or a weak transmission barrier. In contrast, a high species barrier was observed for the non-BSE related prions that harboured an identical PrP amino acid sequence such as sheep-scrapie, mouse RML or human sCJD isolates, supporting the theory that the prion transmission barrier is modulated by strain properties (presumably conformation-dependent) rather than by PrP amino acid sequence differences between host and donor.


As identical results were observed with prions propagated either in natural hosts or in transgenic mouse models, we postulate that the species barrier and its passage consequences are uniquely governed by the host PrPC sequence and not influenced by the PrPC expression level or genetic factors other than the PrPC amino acid sequence. All these findings unequivocally demonstrate that the species barrier and its passage consequences are uniquely driven by the PrPC sequence, and not by other host genetic factors, demonstrating the validity of transgenic PrP animals as models for studies of the species barrier.


The results presented herein reinforce the idea that the BSE agent is highly promiscuous, infecting other species, maintaining its properties in the new species, and even increasing its capabilities to jump to other species including humans. These data are essential for the development of an accurate risk assessment for BSE.


P.150: Zoonotic potential of L-type BSE prions: A new prion disease in humans?


Emilie Jaumain,1 Stéphane Haïk,2 Isabelle Quadrio,3 Laetitia Herzog,1 Fabienne Reine,1 Armand Perret-Liaudet,3 Human Rezaei,1 Hubert Laude,1 Jean-Luc Vilotte,4 and Vincent Béringue1 1INR A (Institut National de la Recherche Agronomique); UR892; Virologie Immunologie Moléculaires; Jouy-en-Josas, France; 2IN SERM; Equipe maladie d’Alzheimer et maladies à Prions; CRicm; UMRS 1127; CNR S; UPMC. R.; ICM, Hôpital de la Salpêtrière; Paris, France; 3Neurobiologie, CMRR , Gériatrie, Hospices Civils de Lyon, Université Lyon 1-CNR S UMR5292-IN SERM U1028; Lyon, France; 3INR A; UMR1313; Génétique Animale et Biologie Intégrative; Jouy-en-Josas, France


Two novel prion strains, referred to as BSE-L and BSE-H, have been recognized in bovines through active prion surveillance programs, both being distinct from the epizootic, ‘classical’, BSE strain (C-BSE). Both H and L-types have been detected worldwide as rare cases occurring in aged animals. Like C-BSE prions, H- and L-types prions can propagate with relative ease in foreign species or in transgenic mouse lines expressing heterologous PrP sequences. A prion exhibiting biological properties similar to C-BSE agent sometimes emerged from these cross-species transmissions. Previously, L-type prions were shown to transmit to transgenic mice expressing human PrP with methionine at codon 129 with higher efficacy than C-BSE prions. Here, we examined whether L-type prions propagate without any apparent transmission barrier in these mice and whether such ‘humanised’ L-type prions share biological properties with CJD prions. L-type prions and a panel of human CJD cases with various genotypes at codon 129 and electrophoretic PrPres signatures were serially transmitted by intracerebral route to human PrP mice. The biological phenotypes induced by these agents were compared by all the standard methods currently used to distinguish between prion strains. At each passage, L-type prions were also transmitted back to bovine PrP mice to assess whether the agent has evolved upon passaging on the human PrP sequence. L-type prions transmitted to human PrP mice at 100% attack rate, without notable alteration in the mean incubation times over 5 passages. At each passage, ‘humanized’ L-type prions were able to transmit back to bovine PrP transgenic mice without apparent transmission barrier, as based on the survival time and the restoration of a L-type BSE phenotype. Comparison of mean incubation times on primary and subsequent passages in human PrP mice showed no overlap between L-type and sporadic CJD agents. While the electrophoretic signature and regional distribution of PrPres in L-type diseased mouse brains resembled that seen after transmission of MM2 CJD strain type, both agents exhibited distinct resistance of the associated PrPres molecules to protease denaturation.


In summary, L-type prions can be passaged on the human PrP sequence without any obvious transmission barrier. The phenotype obtained differs from the classical CJD prion types known so far. Careful extrapolation would suggest that the zoonotic transmission of this agent could establish a new prion disease type in humans.



Protein Structure and Biology



P.200: Clinical expression of BSE in mouse models is unrelated to hallmarks of prion diseases


Christelle Jas-Duval,1,2 Jacqueline Mikol,1 Sophie Luccantoni-Freire,1 Christine Defer,2 Jean-Jacques Huart,2 Paul Brown,1 Jean-Philippe Deslys,1 and Emmanuel E Comoy1 1CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), Division of Prions and Related Diseases (SEPIA); Fontenay-aux-Roses, France; 2EFS-Nord de France; Lille, France


Background. In 1997, we demonstrated that intracerebral inoculation of cattle BSE to C57Bl/6 mice induced neurological disease, even though half of the recipient animals were devoid of PrPres (Lasmezas et al., 1997). PrPres appeared with shortening incubation periods after subsequent passages, suggesting that detectable PrPres should be considered as an indicator of strain virulence rather than as a specific (and required) marker of prion diseases. Subsequent blood risk studies showed that cynomolgus macaques exposed to vCJD-contaminated blood products developed either classical vCJD or a novel neurological disease without detectable PrPres, In our continuing studies of this phenomenon, we now report the results of intravenous inoculations of several different wild-type mouse strains with bovine or primate BSE source material.


Materials and Methods. Brains from one bovine and one cynomolgus monkey showing clinical signs of BSE were sonicated and ultracentrifuged at 188,000 g for 1 hour. Resulting pellets and supernatants (equivalent to 2 mg of brain / mouse) were intravenously injected to PrP+/+ Swiss, PrP+/- and PrP+/+ C57Bl/6N mice, totalling 12 separate bioassays. PrPres was detected using conventional ELISA, western blotting and IHC procedures. Pathology was studied on formalin-fixed brain tissues.


Results. The transmission rate of BSE in those 12 experiments ranged from 0% (PrP+/- C57Bl/6N—cattle BSE—supernatant or pellet: no clinical sign, no spongiosis and no PrPres) to 100% (Swiss—primate BSE—supernatant or pellet: all mice exhibited clinical signs, spongiosis and PrPres). Overall, transmission was more efficient in (1) Swiss than C57Bl/6 mice, (2) PrP+/+ than PrP+/- mice, (3) primate than cattle BSE and (4) pellets than supernatant preparations. Among the 8 models exhibiting partial transmission ratios, 29 mice showed clinical neurological signs, of which only one-third (10) had detectable spongiosis and PrPres. The other animals exhibited only PrPres (7), only spongiosis (5) or neither (7).


Conclusion. Our results suggest that clinical neurotoxicity, spongiosis, and accumulation of PrPres are three interconnected but disparate phenomena. Spongiosis and PrPres are specific but not systematic hallmarks of the onset of prion diseases, notably upon first passage of non-adapted prion strains in a new host. These results question the universality of current human diagnostic criteria and the real prevalence of disease linked to BSE exposure.



***In addition, non-human primates are specifically susceptible for atypical BSE as demonstrated by an approximately 50% shortened incubation time for L-type BSE as compared to C-type. Considering the current scientific information available, it cannot be assumed that these different BSE types pose the same human health risks as C-type BSE or that these risks are mitigated by the same protective measures.



BSE prions propagate as either variant CJD-like or sporadic CJD-like prion strains in transgenic mice expressing human prion protein


*** Surprisingly, however, BSE transmission to these transgenic mice, in addition to producing a vCJD-like phenotype, can also result in a distinct molecular phenotype that is indistinguishable from that of sporadic CJD with PrPSc type 2.


These data suggest that more than one BSEderived prion strain might infect humans;


***it is therefore possible that some patients with a phenotype consistent with sporadic CJD may have a disease arising from BSE exposure.




These studies further strengthen the evidence that vCJD is caused by a BSE-like prion strain.


Also, remarkably, the key neuropathological hallmark of vCJD, the presence of abundant florid PrP plaques, can be recapitulated on BSE or vCJD transmission to these mice.


***However, the most surprising aspect of the studies was the finding that an alternate pattern of disease can be induced in 129MM Tg35 mice from primary transmission of BSE, with a molecular phenotype indistinguishable from that of a subtype of sporadic CJD. This finding has important potential implications as it raises the possibility that some humans infected with BSE prions may develop a clinical disease indistinguishable from classical CJD associated with type 2 PrPSc. This is, in our experience, the commonest molecular sub-type of sporadic CJD. In this regard, it is of interest that the reported incidence of sporadic CJD has risen in the UK since the 1970s (Cousens et al., 1997)...



To date the OIE/WAHO assumes that the human and animal health standards set out in the BSE chapter for classical BSE (C-Type) applies to all forms of BSE which include the H-type and L-type atypical forms. This assumption is scientifically not completely justified and accumulating evidence suggests that this may in fact not be the case. Molecular characterization and the spatial distribution pattern of histopathologic lesions and immunohistochemistry (IHC) signals are used to identify and characterize atypical BSE. Both the L-type and H-type atypical cases display significant differences in the conformation and spatial accumulation of the disease associated prion protein (PrPSc) in brains of afflicted cattle. Transmission studies in bovine transgenic and wild type mouse models support that the atypical BSE types might be unique strains because they have different incubation times and lesion profiles when compared to C-type BSE. When L-type BSE was inoculated into ovine transgenic mice and Syrian hamster the resulting molecular fingerprint had changed, either in the first or a subsequent passage, from L-type into C-type BSE.


***In addition, non-human primates are specifically susceptible for atypical BSE as demonstrated by an approximately 50% shortened incubation time for L-type BSE as compared to C-type. Considering the current scientific information available, it cannot be assumed that these different BSE types pose the same human health risks as C-type BSE or that these risks are mitigated by the same protective measures.



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


Subject: re-BSE prions propagate as either variant CJD-like or sporadic CJD


Date: Thu, 28 Nov 2002 10:23:43 -0000


From: "Asante, Emmanuel A" e.asante@ic.ac.uk


To: "'flounder@wt.net'" flounder@wt.net


Dear Terry,


I have been asked by Professor Collinge to respond to your request. I am a Senior Scientist in the MRC Prion Unit and the lead author on the paper. I have attached a pdf copy of the paper for your attention.


Thank you for your interest in the paper.


In respect of your first question, the simple answer is, ***yes. As you will find in the paper, we have managed to associate the alternate phenotype to type 2 PrPSc, the commonest sporadic CJD. It is too early to be able to claim any further sub-classification in respect of Heidenhain variant CJD or Vicky Rimmer's version. It will take further studies, which are on-going, to establish if there are sub-types to our initial finding which we are now reporting. The main point of the paper is that, as well as leading to the expected new variant CJD phenotype, BSE transmission to the 129-methionine genotype can lead to an alternate phenotype which is indistinguishable from type 2 PrPSc.


I hope reading the paper will enlighten you more on the subject. If I can be of any further assistance please to not hesitate to ask. Best wishes.


Emmanuel Asante






Dr. Emmanuel A Asante MRC Prion Unit & Neurogenetics Dept. Imperial College School of Medicine (St. Mary's) Norfolk Place, LONDON W2 1PG Tel: +44 (0)20 7594 3794 Fax: +44 (0)20 7706 3272 email: e.asante@ic.ac.uk (until 9/12/02) New e-mail: e.asante@prion.ucl.ac.uk (active from now)




Thursday, August 12, 2010


Seven main threats for the future linked to prions


First threat


The TSE road map defining the evolution of European policy for protection against prion diseases is based on a certain numbers of hypotheses some of which may turn out to be erroneous. In particular, a form of BSE (called atypical Bovine Spongiform Encephalopathy), recently identified by systematic testing in aged cattle without clinical signs, may be the origin of classical BSE and thus potentially constitute a reservoir, which may be impossible to eradicate if a sporadic origin is confirmed.


***Also, a link is suspected between atypical BSE and some apparently sporadic cases of Creutzfeldt-Jakob disease in humans. These atypical BSE cases constitute an unforeseen first threat that could sharply modify the European approach to prion diseases.


Second threat





Monday, October 10, 2011


EFSA Journal 2011 The European Response to BSE: A Success Story




EFSA and the European Centre for Disease Prevention and Control (ECDC) recently delivered a scientific opinion on any possible epidemiological or molecular association between TSEs in animals and humans (EFSA Panel on Biological Hazards (BIOHAZ) and ECDC, 2011). This opinion confirmed Classical BSE prions as the only TSE agents demonstrated to be zoonotic so far


*** but the possibility that a small proportion of human cases so far classified as "sporadic" CJD are of zoonotic origin could not be excluded. Moreover, transmission experiments to non-human primates suggest that some TSE agents in addition to Classical BSE prions in cattle (namely L-type Atypical BSE, Classical BSE in sheep, transmissible mink encephalopathy (TME) and chronic wasting disease (CWD) agents) might have zoonotic potential.









***Moreover, L-BSE has been transmitted more easily to transgenic mice overexpressing a human PrP [13,14] or to primates [15,16] than C-BSE.


***It has been suggested that some sporadic CJD subtypes in humans may result from an exposure to the L-BSE agent. Lending support to this hypothesis, pathological and biochemical similarities have been observed between L-BSE and an sCJD subtype (MV genotype at codon 129 of PRNP) [17], and between L-BSE infected non-human primate and another sCJD subtype (MM genotype) [15].






Monday, June 23, 2014





Bovine Spongiform Encephalopathy BSE (typical and atypical strains) USDA


*** Saturday, November 2, 2013 ***


Exploring the risks of a putative transmission of BSE to new species



Wednesday, September 25, 2013


Presence of subclinical infection in gene-targeted human prion protein transgenic mice exposed to atypical BSE



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 ;




*** ''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




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



Saturday, August 14, 2010 BSE


Case Associated with Prion Protein Gene Mutation (g-h-BSEalabama) and VPSPr PRIONPATHY



Sunday, September 1, 2013


*** Evaluation of the Zoonotic Potential of Transmissible Mink Encephalopathy


We previously described the biochemical similarities between PrPres derived from L-BSE infected macaque and cortical MM2 sporadic CJD: those observations suggest a link between these two uncommon prion phenotypes in a primate model (it is to note that such a link has not been observed in other models less relevant from the human situation as hamsters or transgenic mice overexpressing ovine PrP [28]). We speculate that a group of related animal prion strains (L-BSE, c-BSE and TME) would have a zoonotic potential and lead to prion diseases in humans with a type 2 PrPres molecular signature (and more specifically type 2B for vCJD)




Together with previous experiments performed in ovinized and bovinized transgenic mice and hamsters [8,9] indicating similarities between TME and L-BSE, the data support the hypothesis that L-BSE could be the origin of the TME outbreaks in North America and Europe during the mid-1900s.



Saturday, November 2, 2013


*** APHIS Finalizes Bovine Import Regulations in Line with International Animal Health Standards while enhancing the spread of BSE TSE prion mad cow type disease around the Globe ***





Owens, Julie


From: Terry S. Singeltary Sr. [flounder9@verizon.net]


Sent: Monday, July 24, 2006 1:09 PM


To: FSIS RegulationsComments


Subject: [Docket No. FSIS-2006-0011] FSIS Harvard Risk Assessment of Bovine Spongiform Encephalopathy (BSE) Page 1 of 98






*** Singeltary reply ; Molecular, Biochemical and Genetic Characteristics of BSE in Canada Singeltary reply ;





Date: March 21, 2007 at 2:27 pm PST






Bulk cattle feed made with recalled Darling's 85% Blood Meal, Flash Dried, Recall # V-024-2007




Cattle feed delivered between 01/12/2007 and 01/26/2007




Pfeiffer, Arno, Inc, Greenbush, WI. by conversation on February 5, 2007.


Firm initiated recall is ongoing.




Blood meal used to make cattle feed was recalled because it was cross- contaminated with prohibited bovine meat and bone meal that had been manufactured on common equipment and labeling did not bear cautionary BSE statement.




42,090 lbs.














The firm does not utilize a code - only shipping documentation with commodity and weights identified.




Rangen, Inc, Buhl, ID, by letters on February 13 and 14, 2007. Firm initiated recall is complete.




Products manufactured from bulk feed containing blood meal that was cross contaminated with prohibited meat and bone meal and the labeling did not bear cautionary BSE statement.




9,997,976 lbs.




ID and NV





Sunday, December 15, 2013





Tuesday, August 12, 2014





Thursday, June 23, 2011


Experimental H-type bovine spongiform encephalopathy characterized by plaques and glial- and stellate-type prion protein deposits



Wednesday, April 25, 2012






Saturday, August 4, 2012


*** Final Feed Investigation Summary - California BSE Case - July 2012 (ATYPICAL L-TYPE BASE BSE)



Discussion: The C, L and H type BSE cases in Canada exhibit molecular characteristics similar to those described for classical and atypical BSE cases from Europe and Japan. *** This supports the theory that the importation of BSE contaminated feedstuff is the source of C-type BSE in Canada. *** It also suggests a similar cause or source for atypical BSE in these countries. ***


see page 176 of 201 pages...tss



*** Singeltary reply ; Molecular, Biochemical and Genetic Characteristics of BSE in Canada Singeltary reply ;



I think they still test the lowest possible by oie standards, to keep them in line for trade, which is about 40,000 a year, from 95 million or so...tss




Why is USDA "only" testing 40,000 samples a year? USDA's surveillance strategy is to focus on the targeted populations where we are most likely to find disease if it is present. This is the most effective way to meet both OIE and our domestic surveillance standards. After completing our enhanced surveillance in 2006 and confirming that our BSE prevalence was very low, we concluded that 40,000 samples per year from these targeted, high risk populations would far exceed these standards. In fact, this sampling is ten times greater than OIE standards .



The total number of cattle and calves in the U.S. on January 1, 2014, was 87.73 million head, down 1.8% from January 2013 and 9.2% lower than at the last cyclical peak in 2007. This is the lowest January cattle inventory since 1951, _supposedly_, due to the drought, but I have my concern with that. the 2004 enhanced BSE surveillance program was shut down after finding two mad cows (atypical's), and in my opinion, that's why they shut it down. also, Dr. Paul Brown from the cdc said he trusted nothing from them after the Texas 2nd mad cow was covered up for months and months, on what was suppose to be a 48 hour turn around on testing. he said this ;


"Everything they did on the Texas cow makes everything USDA did before 2005 suspect," Brown said. (see link below).


the rest is history ;


An Arizona meat processing company and its owner pled guilty in February 2007 to charges of theft of Government funds, mail fraud, and wire fraud. The owner and his company defrauded the BSE Surveillance Program when they falsified BSE Surveillance Data Collection Forms and then submitted payment requests to USDA for the services. In addition to the targeted sample population (those cattle that were more than 30 months old or had other risk factors for BSE), the owner submitted to USDA, or caused to be submitted, BSE obex (brain stem) samples from healthy USDA-inspected cattle. As a result, the owner fraudulently received approximately $390,000. Sentencing is scheduled for May 2007.




Topics that will be covered in ongoing or planned reviews under Goal 1 include:


soundness of BSE maintenance sampling (APHIS),


implementation of Performance-Based Inspection System enhancements for specified risk material (SRM) violations and improved inspection controls over SRMs (FSIS and APHIS),




The findings and recommendations from these efforts will be covered in future semiannual reports as the relevant audits and investigations are completed.





Office of the United States Attorney District of Arizona FOR IMMEDIATE RELEASE For Information Contact Public Affairs February 16, 2007 WYN HORNBUCKLE Telephone: (602) 514-7625 Cell: (602) 525-2681




PHOENIX -- Farm Fresh Meats, Inc. and Roland Emerson Farabee, 55, of Maricopa, Arizona, pleaded guilty to stealing $390,000 in government funds, mail fraud and wire fraud, in federal district court in Phoenix. U.S. Attorney Daniel Knauss stated, “The integrity of the system that tests for mad cow disease relies upon the honest cooperation of enterprises like Farm Fresh Meats. Without that honest cooperation, consumers both in the U.S. and internationally are at risk.



WE can only hope that this is a single incident. BUT i have my doubts. I remember when the infamous TOKEN Purina Feed Mill in Texas was feeding up to 5.5 grams of potentially and probably tainted BANNED RUMINANT feed to cattle, and the FDA was bragging at the time that the amount of potentially BANNED product was so little and the cattle were so big ;


"It is important to note that the prohibited material was domestic in origin (therefore not likely to contain infected material because there is no evidence of BSE in U.S. cattle), fed at a very low level, and fed only once. The potential risk of BSE to such cattle is therefore exceedingly low, even if the feed were contaminated."



On Friday, April 30 th , the Food and Drug Administration learned that a cow with central nervous system symptoms had been killed and shipped to a processor for rendering into animal protein for use in animal feed. ... FDA's investigation showed that the animal in question had already been rendered into "meat and bone meal" (a type of protein animal feed). Over the weekend FDA was able to track down all the implicated material. That material is being held by the firm, which is cooperating fully with FDA.



WE now know all that was a lie. WE know that literally Thousands of TONS of BANNED and most likely tainted product is still going out to commerce. WE know now and we knew then that .005 to a gram was lethal. WE know that CWD infected deer and elk, scrapie infected sheep, BSE and BASE infected cattle have all been rendered and fed back to livestock (including cattle) for human and animal consumption.


Paul Brown, known and respected TSE scientist, former TSE expert for the CDC said he had ''absolutely no confidence in USDA tests before one year ago'', and this was on March 15, 2006 ;


"The fact the Texas cow showed up fairly clearly implied the existence of other undetected cases," Dr. Paul Brown, former medical director of the National Institutes of Health's Laboratory for Central Nervous System Studies and an expert on mad cow-like diseases, told United Press International. "The question was, 'How many?' and we still can't answer that."


Brown, who is preparing a scientific paper based on the latest two mad cow cases to estimate the maximum number of infected cows that occurred in the United States, said he has "absolutely no confidence in USDA tests before one year ago" because of the agency's reluctance to retest the Texas cow that initially tested positive.


USDA officials finally retested the cow and confirmed it was infected seven months later, but only at the insistence of the agency's inspector general.


"Everything they did on the Texas cow makes everything USDA did before 2005 suspect," Brown said. ...snip...end



CDC - Bovine Spongiform Encephalopathy and Variant Creutzfeldt ... Dr. Paul Brown is Senior Research Scientist in the Laboratory of Central Nervous System ... Address for correspondence: Paul Brown, Building 36, Room 4A-05, ...





Tuesday, September 12, 2006 11:10 AM


"Actually, Terry, I have been critical of the USDA handling of the mad cow issue for some years, and with Linda Detwiler and others sent lengthy detailed critiques and recommendations to both the USDA and the Canadian Food Agency."


OR, what the Honorable Phyllis Fong of the OIG found ;


Finding 2 Inherent Challenges in Identifying and Testing High-Risk Cattle Still Remain



*** Singeltary reply ; Molecular, Biochemical and Genetic Characteristics of BSE in Canada Singeltary reply ;



Tuesday, August 12, 2014





Thursday, October 02, 2014


[Docket No. APHIS-2013-0064] Concurrence With OIE Risk Designations for Bovine Spongiform Encephalopathy



Saturday, August 14, 2010


BSE Case Associated with Prion Protein Gene Mutation (g-h-BSEalabama) and VPSPr PRIONPATHY






Transmissible Spongiform Encephalopathy TSE Prion Disease North America 2014


Transmissible Spongiform Encephalopathy TSE Prion Disease have now been discovered in a wide verity of species across North America. typical C-BSE, atypical L-type BASE BSE, atypical H-type BSE, atypical H-G BSE, of the bovine, typical and atypical Scrapie strains, in sheep and goats, with atypical Nor-98 Scrapie spreading coast to coast in about 5 years. Chronic Wasting Disease CWD in cervid is slowly spreading without any stopping it in Canada and the USA and now has mutated into many different strains. Transmissible Mink Encephalopathy TME outbreaks. These Transmissible Spongiform Encephalopathy TSE Prion Disease have been silently mutating and spreading in different species in North America for decades.


The USDA, FDA, et al have assured us of a robust Triple BSE TSE prion Firewall, of which we now know without a doubt, that it was nothing but ink on paper. Since the 1997 mad cow feed ban in the USA, literally tons and tons of banned mad cow feed has been put out into commerce, never to return, as late as December of 2013, serious, serious breaches in the FDA mad cow feed ban have been documented. The 2004 enhanced BSE surveillance program was so flawed, that one of the top TSE prion Scientist for the CDC, Dr. Paul Brown stated ; Brown, who is preparing a scientific paper based on the latest two mad cow cases to estimate the maximum number of infected cows that occurred in the United States, said he has "absolutely no confidence in USDA tests before one year ago" because of the agency's reluctance to retest the Texas cow that initially tested positive. see ; http://www.upi.com/Health_News/2006/03/15/Analysis-What-that-mad-cow-means/UPI-12841142465253/


The BSE surveillance and testing have also been proven to be flawed, and the GAO and OIG have both raised serious question as to just how flawed it has been (see GAO and OIG reports). North America has more documented TSE prion disease, in different documented species (excluding the Zoo BSE animals in the EU), then any other place on the Globe. This does not include the very likelihood that TSE prion disease in the domestic feline and canine have been exposed to high doses of the TSE prion disease vid pet food. To date, it’s still legal to include deer from cwd zone into pet food or deer food. Specified Risk Material i.e. SRM bans still being breach, as recently as just last month.


nvCJD or what they now call vCJD, another case documented in Texas last month, with very little information being released to the public on about this case? with still the same line of thought from federal officials, ‘it can’t happen here’, so another vCJD blamed on travel of a foreign animal disease from another country, while ignoring all the BSE TSE Prion risk factors we have here in the USA and Canada, and the time that this victim and others, do spend in the USA, and exposed to these risk factors, apparently do not count in any way with regard to risk factor. a flawed process of risk assessment.


sporadic CJD, along with new TSE prion disease in humans, of which the young are dying, of which long duration of illness from onset of symptoms to death have been documented, only to have a new name added to the pot of prion disease i.e. sporadic GSS, sporadic FFI, and or VPSPR. I only ponder how a familial type disease could be sporadic with no genetic link to any family member? when the USA is the only documented Country in the world to have documented two different cases of atypical H-type BSE, with one case being called atypical H-G BSE with the G meaning Genetic, with new science now showing that indeed atypical H-type BSE is very possible transmitted to cattle via oral transmission (Prion2014). sporadic CJD and VPSPR have been rising in Canada, USA, and the UK, with the same old excuse, better surveillance. You can only use that excuse for so many years, for so many decades, until one must conclude that CJD TSE prion cases are rising. a 48% incease in CJD in Canada is not just a blip or a reason of better surveillance, it is a mathematical rise in numbers. More and more we are seeing more humans exposed in various circumstance in the Hospital, Medical, Surgical arenas to the TSE Prion disease, and at the same time in North America, more and more humans are becoming exposed to the TSE prion disease via consumption of the TSE prion via deer and elk, cattle, sheep and goats, and for those that are exposed via or consumption, go on to further expose many others via the iatrogenic modes of transmission of the TSE prion disease i.e. friendly fire. I pondered this mode of transmission via the victims of sporadic FFI, sporadic GSS, could this be a iatrogenic event from someone sub-clinical with sFFI or sGSS ? what if?


Two decades have passed since Dr. Ironside first confirmed his first ten nvCJD victims in 1995. Ten years later, 2005, we had Dr. Gambetti and his first ten i.e. VPSPR in younger victims. now we know that indeed VPSPR is transmissible. yet all these TSE prion disease and victims in the USA and Canada are being pawned off as a spontaneous event, yet science has shown, the spontaneous theory has never been proven in any natural case of TSE prion disease, and scientist have warned, that they have now linked some sporadic CJD cases to atypical BSE, to atypical Scrapie, and to CWD, yet we don’t here about this in the public domain. We must make all human and animal TSE prion disease reportable in every age group, in ever state and internationally, we must have a serious re-evaluation and testing of the USA cattle herds, and we must ban interstate movement of all cervids. Any voluntary effort to do any of this will fail. Folks, we have let the industry run science far too long with regards to the TSE prion disease. While the industry and their lobbyist continues to funnel junk science to our decision policy makers, Rome burns. ...end




[all scientific peer review studies and other scientific information I have put into blogs, to shorten reference data. I DO NOT advertise or make money from this, this information is for education use...lost my mom to the hvCJD, and just made a promise, never forget, and never let them forget. ...TSS]


Sunday, June 29, 2014


Transmissible Spongiform Encephalopathy TSE Prion Disease North America 2014



Monday, November 3, 2014




National Prion Disease Pathology Surveillance Center Cases Examined1 (October 7, 2014)


***6 Includes 11 cases in which the diagnosis is pending, and 19 inconclusive cases; ***7 Includes 12 (11 from 2014) cases with type determination pending in which the diagnosis of vCJD has been excluded. ***The sporadic cases include 2660 cases of sporadic Creutzfeldt-Jakob disease (sCJD),


***50 cases of Variably Protease-Sensitive Prionopathy (VPSPr)


***and 21 cases of sporadic Fatal Insomnia (sFI).



Monday, November 3, 2014


The prion protein protease sensitivity, stability and seeding activity in variably protease sensitive prionopathy brain tissue suggests molecular overlaps with sporadic Creutzfeldt-Jakob disease




Saturday, August 14, 2010 BSE


Case Associated with Prion Protein Gene Mutation (g-h-BSEalabama) and VPSPr PRIONPATHY







Monday, June 02, 2014 Confirmed Variant CJD Case in Texas



SO, 4 months after the fact and still no word on this case. no information what so ever. the silence is deafening $$$