Atypical BSE, BSE, and other human and animal TSE in North America Update October 19, 2009

October 19, 2009

Greetings,

An update of sorts on atypical BSE and other TSE in North America, reported, and or, not reported. Please remember, the _typical_ U.K. c-BSE, the l-BSE (BASE), and the h-BSE have all been documented in North America, along with the typical scrapie's, and atypical Nor-98 Scrapie, and to date, 2 different strains of CWD, and also TME. please remember, all these TSE in different species have been rendered and feed to food producing animals for humans and animals in North America (absolutely no idea of TSE in cats and dogs see reference on that), and that the trading of these TSEs via animals and products via the USA and Canada has been so immense over the years, decades, that it was like swapping spit between two lovers. also, please remember, in my opinion (I will show the facts to prove this), Canada is Looking to find TSE in cattle, and the USA has done just the opposite, the look NOT to find and report. The SSS policy has been in full force in the USA for some time. also, there will be some additional information on Transmission studies. Also, what about any human TSE there from, and the surveillance there of ???

With that said, I present you with these facts as follows. There is new data mixed up with old data, so don't miss any of it. ...kind regards, terry

Wednesday, February 11, 2009

Atypical BSE North America Update February 2009 Both of the BSE cases ascertained in the US native-born cattle were atypical cases (H-type), which contributed to the initial ambiguity of the diagnosis. 174, 185 In Canada, there have been 2 atypical BSE cases in addition to the 14 cases of the classic UK strain of BSE2: one was the H-type, and the other was of the L-type.198

snip...end

source : Enhanced Abstract Journal of the American Veterinary Medical Association January 1, 2009, Vol. 234, No. 1, Pages 59-72 Bovine spongiform encephalopathy Jane L. Harman, DVM, PhD; Christopher J. Silva, PhD


http://avmajournals.avma.org/doi/ref/10.2460/javma.234.1.59




October 2009

O.11.3

Infectivity in skeletal muscle of BASE-infected cattle

Silvia Suardi1, Chiara Vimercati1, Fabio Moda1, Ruggerone Margherita1, Ilaria Campagnani1, Guerino Lombardi2, Daniela Gelmetti2, Martin H. Groschup3, Anne Buschmann3, Cristina Casalone4, Maria Caramelli4, Salvatore Monaco5, Gianluigi Zanusso5, Fabrizio Tagliavini1 1Carlo Besta” Neurological Institute,Italy; 2IZS Brescia, Italy; 33FLI Insel Riems, D, Germany; 4CEA-IZS Torino, Italy; 5University of Verona, Italy

Background: BASE is an atypical form of bovine spongiform encephalopathy caused by a prion strain distinct from that of BSE. Upon experimental transmission to cattle, BASE induces a previously unrecognized disease phenotype marked by mental dullness and progressive atrophy of hind limb musculature. Whether affected muscles contain infectivity is unknown. This is a critical issue since the BASE strain is readily transmissible to a variety of hosts including primates, suggesting that humans may be susceptible.

Objectives: To investigate the distribution of infectivity in peripheral tissues of cattle experimentally infected with BASE. Methods: Groups of Tg mice expressing bovine PrP (Tgbov XV, n= 7-15/group) were inoculated both i.c. and i.p. with 10% homogenates of a variety of tissues including brain, spleen, cervical lymph node, kidney and skeletal muscle (m. longissimus dorsi) from cattle intracerebrally infected with BASE. No PrPres was detectable in the peripheral tissues used for inoculation either by immunohistochemistry or Western blot.

Results: Mice inoculated with BASE-brain homogenates showed clinical signs of disease with incubation and survival times of 175±15 and 207±12 days. Five out of seven mice challenged with skeletal muscle developed a similar neurological disorder, with incubation and survival times of 380±11 and 410±12 days. At present (700 days after inoculation) mice challenged with the other peripheral tissues are still healthy. The neuropathological phenotype and PrPres type of the affected mice inoculated either with brain or muscle were indistinguishable and matched those of Tgbov XV mice infected with natural BASE.

Discussion: Our data indicate that the skeletal muscle of cattle experimentally infected with BASE contains significant amount of infectivity, at variance with BSE-affected cattle, raising the issue of intraspecies transmission and the potential risk for humans. Experiments are in progress to assess the presence of infectivity in skeletal muscles of natural BASE.

P.4.23

Transmission of atypical BSE in humanized mouse models

Liuting Qing1, Wenquan Zou1, Cristina Casalone2, Martin Groschup3, Miroslaw Polak4, Maria Caramelli2, Pierluigi Gambetti1, Juergen Richt5, Qingzhong Kong1 1Case Western Reserve University, USA; 2Instituto Zooprofilattico Sperimentale, Italy; 3Friedrich-Loeffler-Institut, Germany; 4National Veterinary Research Institute, Poland; 5Kansas State University (Previously at USDA National Animal Disease Center), USA

Background: Classical BSE is a world-wide prion disease in cattle, and the classical BSE strain (BSE-C) has led to over 200 cases of clinical human infection (variant CJD). Atypical BSE cases have been discovered in three continents since 2004; they include the L-type (also named BASE), the H-type, and the first reported case of naturally occurring BSE with mutated bovine PRNP (termed BSE-M). The public health risks posed by atypical BSE were largely undefined.

Objectives: To investigate these atypical BSE types in terms of their transmissibility and phenotypes in humanized mice.

Methods: Transgenic mice expressing human PrP were inoculated with several classical (C-type) and atypical (L-, H-, or Mtype) BSE isolates, and the transmission rate, incubation time, characteristics and distribution of PrPSc, symptoms, and histopathology were or will be examined and compared.

Results: Sixty percent of BASE-inoculated humanized mice became infected with minimal spongiosis and an average incubation time of 20-22 months, whereas only one of the C-type BSE-inoculated mice developed prion disease after more than 2 years. Protease-resistant PrPSc in BASE-infected humanized Tg mouse brains was biochemically different from bovine BASE or sCJD. PrPSc was also detected in the spleen of 22% of BASE-infected humanized mice, but not in those infected with sCJD. Secondary transmission of BASE in the humanized mice led to a small reduction in incubation time. The atypical BSE-H strain is also transmissible with distinct phenotypes in the humanized mice, but no BSE-M transmission has been observed so far.

Discussion: Our results demonstrate that BASE is more virulent than classical BSE, has a lymphotropic phenotype, and displays a modest transmission barrier in our humanized mice. 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.

P.5.3

Differences in the expression levels of selected genes in the brain tissue of cattle naturally infected with classical and atypical BSE.

Magdalena Larska1, Miroslaw P. Polak1, Jan F. Zmudzinski1, Juan M. Torres2 1National Veterinary Institute, Poland; 2CISA/INIA

Background: Recently cases of BSE in older cattle named BSE type L and type H were distinguished on the basis of atypical glycoprofiles of PrPres. The nature of those strains is still not fully understood but it is suspected that the atypical BSE cases are sporadic. Hitherto most BSE cases were studied in respect to the features of PrPSc. Here we propose gene expression profiling as a method to characterize and distinguish BSE strains.

Objectives: The aim of the study was to compare the activities of some factors which are known to play a role in TSE’s pathogenesis in order to distinguish the differences/similarities between all BSE types. Methods: 10 % homogenate of brain stem tissue collected from obex region of medulla oblongata from 20 naturally infected BSE cows (8 assigned as classical BSE, other 8 and 4 infected with atypical BSE L type and H type respectively) was used in the study. As negative control animals we’ve used 8 animals in the age between 2.5 and 13 years. The genes were relatively quantified using SYBR Green real time RT-PCR. Raw data of Ct values was transformed into normalized relative quantities using Qbase Plus®.

Results and Discussion: In most of the tested genes significant differences in the expression levels between the brain stem of healthy cattle and animals infected with different BSE types were observed. In c-type BSE in comparison to healthy and atypical BSE the overexpression of the gene of bcl-2, caspase 3, 14-3-3 and tylosine kinase Fyn was significant. Simultaneously in atypical BSEs type-L and type-H the levels of prion protein, Bax and LPR gene was elevated in comparison to c-BSE. Additionally L-BSE was characterized by the overexpression of STI1 and SOD genes compared to the other of BSE types. The downregulation of the gene encoding NCAM1 was observed in all BSE types in comparison to healthy cows. Different gene expression profiles of bovine brains infected with classical and atypical BSE indicates possible different pathogenesis or source of the disease.

O.10.1

Transmission of uncommon forms of bovine prions to transgenic mice expressing human PrP: questions and progress

Vincent Béringue, Hubert Laude INRA, UR 892, Virologie Immunologie Moléculaires, France

The active, large-scale testing of livestock nervous tissues for the presence of protease-resistant prion protein (PrPres) has led to the recognition of 2 uncommon PrPres molecular signatures, termed H-type and L-type BSE. Their experimental transmission to various transgenic and inbred mouse lines unambiguously demonstrated the infectious nature of such cases and the existence of distinct prion strains in cattle. Like the classical BSE agent, H- and L-type (or BASE) prions can propagate in heterologous species. In addition L-type prions acquire molecular and neuropathologic phenotypic traits undistinguishable from BSE or BSE-related agents upon transmission to transgenic mice expressing ovine PrP (VRQ allele) or wild-type mice. An understanding of the transmission properties of these newly recognized prions when confronted with human PrP sequence was therefore needed. Toward this end, we inoculated mice expressing human PrP Met129 with several field isolates. Unlike classical BSE agent, L-type prions appeared to propagate in these mice with no obvious transmission barrier. In contrast, we repeatedly failed to infect them with Htype prions. Ongoing investigations aim to extend the knowledge on these uncommon strains: are these agents able to colonize lymphoid tissue, a potential key factor for successful transmission by peripheral route; is there any relationship between these assumedly sporadic forms of TSE in cattle and some sporadic forms of human CJD are among the issues that need to be addressed for a careful assessment of the risk for cattle-to-human transmission of H- and L-type prions.

O.4.3

Spread of BSE prions in cynomolgus monkeys (Macaca fascicularis) after oral transmission

Edgar Holznagel1, Walter Schulz-Schaeffer2, Barbara Yutzy1, Gerhard Hunsmann3, Johannes Loewer1 1Paul-Ehrlich-Institut, Federal Institute for Sera and Vaccines, Germany; 2Department of Neuropathology, Georg-August University, Göttingen, Germany, 3Department of Virology and Immunology, German Primate Centre, Göttingen, Germany

Background: BSE-infected cynomolgus monkeys represent a relevant animal model to study the pathogenesis of variant Creutzfeldt-Jacob disease (vCJD).

Objectives: To study the spread of BSE prions during the asymptomatic phase of infection in a simian animal model.

Methods: Orally BSE-dosed macaques (n=10) were sacrificed at defined time points during the incubation period and 7 orally BSE-dosed macaques were sacrificed after the onset of clinical signs. Neuronal and non-neuronal tissues were tested for the presence of proteinase-K-resistant prion protein (PrPres) by western immunoblot and by paraffin-embedded tissue (PET) blot technique.

Results: In clinically diseased macaques (5 years p.i. + 6 mo.), PrPres deposits were widely spread in neuronal tissues (including the peripheral sympathetic and parasympathetic nervous system) and in lymphoid tissues including tonsils. In asymptomatic disease carriers, PrPres deposits could be detected in intestinal lymph nodes as early as 1 year p.i., but CNS tissues were negative until 3 – 4 years p.i. Lumbal/sacral segments of the spinal cord and medulla oblongata were PrPres positive as early as 4.1 years p.i., whereas sympathetic trunk and all thoracic/cervical segments of the spinal cord were still negative for PrPres. However, tonsil samples were negative in all asymptomatic cases.

Discussion: There is evidence for an early spread of BSE to the CNS via autonomic fibres of the splanchnic and vagus nerves indicating that trans-synaptical spread may be a time-limiting factor for neuroinvasion. Tonsils were predominantly negative during the main part of the incubation period indicating that epidemiological vCJD screening results based on the detection of PrPres in tonsil biopsies may mostly tend to underestimate the prevalence of vCJD among humans.

O.4.4

PrPSc distribution pattern in cattle experimentally challenged with H-type and L-type atypical BSE

Anne Buschmann1, Ute Ziegler1, Leila McIntyre2, Markus Keller1, Ron Rogers3, Bob Hills3, Martin H. Groschup1 1Friedrich-Loeffler-Institut, INEID, Germany; 2Faculty of Veterinary Medicine, University of Calgary, Canada; 3Health Canada, Ottawa, Canada

Background: After the detection of two novel BSE forms designated H-type and L-type BSE, the question of the pathogenesis and the agent distribution in cattle affected with these forms was fully open. From initial studies, it was already known that the PrPSc distribution in L-type BSE affected cattle differed from that known for classical BSE (C-type) where the obex region always displays the highest PrPSc concentrations. In contrast in L-type BSE cases, the thalamus and frontal cortex regions showed the highest levels of the pathological prion protein, while the obex region was only weakly involved. No information was available on the distribution pattern in H-type BSE.

Objectives: To analyse the PrPSc and infectivity distribution in cattle experimentally challenged with H-type and L-type BSE.

Methods: We analysed CNS and peripheral tissue samples collected from cattle that were intracranially challenged with Htype (five animals) and L-type (six animals) using a commercial BSE rapid test (IDEXX HerdChek), immunohistochemistry (IHC) and a highly sensitive Western blot protocol including a phosphotungstic acid precipitation of PrPSc (PTA-WB). Samples collected during the preclinical and the clinical stages of the disease were examined. For the detection of BSE infectivity, selected samples were also inoculated into highly sensitive Tgbov XV mice overexpressing bovine prion protein (PrPC).

Results: Analysis of a collection of fifty samples from the peripheral nervous, lymphoreticular, digestive, reproductive, respiratory and musculo-skeletal systems by PTA-WB, IDEXXHerdChek BSE EIA and IHC revealed a general restriction of the PrPSc accumulation to the central nervous system.

Discussion: Our results on the PrPSc distribution in peripheral tissues of cattle affected with H-type and L-type BSE are generally in accordance with what has been known for C-type BSE. Bioassays are ongoing in highly sensitive transgenic mice in order to reveal infectivity.


http://www.prion2009.com/sites/default/files/Prion2009_Book_of_Abstracts.pdf



Research Project: GENETIC AND BIOLOGICAL DETERMINANTS OF RESPIRATORY DISEASE SUSCEPTIBILITY Location: Animal Health Systems Research

Title: Association of a bovine prion gene haplotype with atypical BSE

Author

Clawson, Michael

Submitted to: Meeting Abstract Publication Type: Abstract Publication Acceptance Date: December 2, 2008 Publication Date: January 1, 2009 Citation: Clawson, M.L. 2009. Association of a bovine prion gene haplotype with atypical BSE [abstract]. Plant and Animal Genomes XVII Conference. Abstract No. W091. Available:


http://www.intl-pag.org/17/abstracts/




Technical Abstract: Transmissible spongiform encephalopathies (TSEs), also known as prion diseases, are a class of fatal neurodegenerative disorders that occur in humans, ruminants, cats, and mink. Three distinct TSEs afflict cattle: classical bovine spongiform encephalopathy (BSE), atypical H-type BSE, and atypical L-type BSE. Classical BSE was identified in the 1980s and is acquired by cattle through the consumption of feed contaminated with the infectious prion agent. Atypical BSEs have only recently been recognized as distinct cattle prion diseases and are extremely rare. The full extent of genetic susceptibilities to atypical BSEs is unknown; however, one atypical H-type case identified in the United States (2006) was most likely caused by a genetic mutation in the prion gene, E211K. We have identified an association of a bovine prion DNA haplotype with atypical BSE that is independent of E211K. The haplotype spans a portion of the prion gene that includes part of intron 2, the entire coding region of exon 3, and part of the three prime untranslated region of exon 3 (13 kb). Despite the low frequency of this haplotype among general cattle populations, it was present in a majority of H- and L-type atypical BSE cases from Canada, France, and the United States. This result indicates that there is a genetic component to atypical BSE susceptibility in addition to E211K.


http://www.ars.usda.gov/research/publications/publications.htm?SEQ_NO_115=234699




I ask Professor Kong ;

Thursday, December 04, 2008 3:37 PM Subject: RE: re--Chronic Wating Disease (CWD) and Bovine Spongiform Encephalopathies (BSE): Public Health Risk Assessment

''IS the h-BSE more virulent than typical BSE as well, or the same as cBSE, or less virulent than cBSE? just curious.....''

Professor Kong reply ;

.....snip

''As to the H-BSE, we do not have sufficient data to say one way or another, but we have found that H-BSE can infect humans. I hope we could publish these data once the study is complete.

Thanks for your interest.''

Best regards,

Qingzhong Kong, PhD Associate Professor Department of Pathology Case Western Reserve University Cleveland, OH 44106 USA

END...TSS

I look forward to further transmission studies, and a true ENHANCED BSE/atypical BSE surveillance program put forth testing all cattle for human and animal consumption for 5 years. a surveillance program that uses the most sensitive TSE testing, and has the personnel that knows how to use them, and can be trusted. I look forward to a stringent mad cow feed ban being put forth, and then strictly enforced. we need a forced, not voluntary feed ban, an enhanced feed ban at that, especially excluding blood. we need some sort of animal traceability. no more excuses about privacy. if somebody is putting out a product that is killing folks and or has the potential to kill you, then everybody needs to know who they are, and where that product came from. same with hospitals, i think medical incidents in all states should be recorded, and made public, when it comes to something like a potential accidental transmission exposure event. so if someone is out there looking at a place to go have surgery done, if you have several hospitals having these type 'accidental exposure events', than you can go some place else. it only makes sense. somewhere along the road, the consumer lost control, and just had to take whatever they were given, and then charged these astronomical prices. some where along the line the consumer just lost interest, especially on a long incubating disease such as mad cow disease i.e. Transmissible Spongiform Encephalopathy. like i said before, there is much more to the mad cow story than bovines and eating a hamburger, we must start focusing on all TSE in all species. ...TSS

Month Number of Tests

Feb 2009 -- 1,891

Jan 2009 -- 4,620


http://www.aphis.usda.gov/newsroom/hot_issues/bse/surveillance/ongoing_surv_results.shtml




P02.35

Molecular Features of the Protease-resistant Prion Protein (PrPres) in H-type BSE

Biacabe, A-G1; Jacobs, JG2; Gavier-Widén, D3; Vulin, J1; Langeveld, JPM2; Baron, TGM1 1AFSSA, France; 2CIDC-Lelystad, Netherlands; 3SVA, Sweden

Western blot analyses of PrPres accumulating in the brain of BSE-infected cattle have demonstrated 3 different molecular phenotypes regarding to the apparent molecular masses and glycoform ratios of PrPres bands. We initially described isolates (H-type BSE) essentially characterized by higher PrPres molecular mass and decreased levels of the diglycosylated PrPres band, in contrast to the classical type of BSE. This type is also distinct from another BSE phenotype named L-type BSE, or also BASE (for Bovine Amyloid Spongiform Encephalopathy), mainly characterized by a low representation of the diglycosylated PrPres band as well as a lower PrPres molecular mass. Retrospective molecular studies in France of all available BSE cases older than 8 years old and of part of the other cases identified since the beginning of the exhaustive surveillance of the disease in 20001 allowed to identify 7 H-type BSE cases, among 594 BSE cases that could be classified as classical, L- or H-type BSE. By Western blot analysis of H-type PrPres, we described a remarkable specific feature with antibodies raised against the C-terminal region of PrP that demonstrated the existence of a more C-terminal cleaved form of PrPres (named PrPres#2 ), in addition to the usual PrPres form (PrPres #1). In the unglycosylated form, PrPres #2 migrates at about 14 kDa, compared to 20 kDa for PrPres #1. The proportion of the PrPres#2 in cattle seems to by higher compared to the PrPres#1. Furthermore another PK–resistant fragment at about 7 kDa was detected by some more N-terminal antibodies and presumed to be the result of cleavages of both N- and C-terminal parts of PrP. These singular features were maintained after transmission of the disease to C57Bl/6 mice. The identification of these two additional PrPres fragments (PrPres #2 and 7kDa band) reminds features reported respectively in sporadic Creutzfeldt-Jakob disease and in Gerstmann-Sträussler-Scheinker (GSS) syndrome in humans.


http://www.neuroprion.com/pdf_docs/conferences/prion2007/abstract_book.pdf




Atypical BSE North America Update February 2009


http://bse-atypical.blogspot.com/2009/02/atypical-bse-north-america-update.html



http://bse-atypical.blogspot.com/2009/06/l-type-bse-h-type-bse-c-type-bse-ibnc.html




Detection of Pathologic Prion Protein in the Olfactory Bulb of Natural and Experimental Bovine Spongiform Encephalopathy Affected Cattle in Great Britain

Y. H. LEE, M. M. SIMMONS, S. A. C. HAWKINS, Y. I. SPENCER, P. WEBB, M. J. STACK, AND G. A. H. WELLS National Veterinary Research and Quarantine Service, Anyang, Republic of Korea (YHL); and Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom (MMS, SACH, YIS, PW, MJS, GAHW)

Abstract.

To investigate the relative involvement of the olfactory region in classical bovine spongiform encephalopathy (BSE), immunohistochemical labeling of prion protein scrapie (PrPSc) was scored in the brainstem, frontal cerebral cortex, and olfactory bulb of cattle with natural and experimental clinical cases of BSE in Great Britain. The intensity of immunolabeling was greatest in the brainstem, but PrPSc was also detected in the olfactory bulb and the cerebral cortex. A diffuse, nonparticulate labeling, possibly due to abundance of cellular PrP, was consistently observed in the olfactory glomeruli of the cases and negative controls. Involvement of the olfactory bulb in BSE and other naturally occurring TSEs of animals raises speculation as to an olfactory portal of infection or a route of excretion of the prion agent.

Key words: BSE; olfactory bulb; olfactory glomeruli; PrPSc; PrPC.

snip...

Only two cases of atypical BSE have been reported from GB; both were H type and were diagnosed on molecular characterization.10 Detailed neuropathologic description of atypical BSE is limited to the L type, originally named bovine amyloidotic spongiform encephalopathy (BASE).3

snip...

In BASE, in addition to PrP-amyloid plaques in the olfactory bulb, the highest levels of PrPSc were recovered from the thalamus and olfactory regions.2 Also, in both captive and free-ranging mule deer with chronic wasting disease, the olfactory cortex has been found to be among the most severely affected areas of the brain.8 Such patterns may be solely a reflection of selective vulnerability of certain neuroanatomic loci and, in end-stage disease, a reflection of phenotype, but equally, they arouse speculation as to possible olfactory portals of infection or excretion of agent. In human transmissible spongiform encephalopathies (TSEs), PrPSc immunolabeling has been reported in the olfactory tract of a variant Creutzfeldt-Jakob disease patient, and in sporadic Creutzfeldt-Jakob disease (sCJD), selective deposition of PrPSc in olfactory glomeruli, olfactory tracts, and olfactory cortex is recorded.6 In the latter study, PrPSc was also reported in the cilia of olfactory receptor neurons and basal cells of the olfactory epithelium but not in the respiratory epithelium. Detection of PrPSc in an olfactory mucosa biopsy, performed 45 days after disease onset in a sCJD patient, led to the suggestion that the involvement of olfactory epithelium might be an early event in sCJD.6

snip...

The present study demonstrates that, in clinical cases, involvement of the olfactory lobe is a consistent phenotypic feature of classic BSE but, in contrast to BASE,3 it is not preferentially affected compared with the cerebral cortex or brainstem. In studies of the pathogenesis of classical BSE after oral exposure, infectivity has not been demonstrated in nasal mucosa,11 but it has been shown, albeit at low titer, in this tissue in clinical cases of scrapie of sheep and goats.5 The detection of PrPSc in olfactory bulb in this study and indeed in other TSEs in terminal disease suggests that olfactory pathways cannot be excluded as a secondary or ancillary route of infection.


http://www.vetpathology.org/cgi/content/abstract/46/1/59



http://www.vetpathology.org/cgi/reprint/46/1/59




???$$$???

Research Project: Study of Atypical Bse Location: Virus and Prion Diseases of Livestock

Project Number: 3625-32000-086-05 Project Type: Specific Cooperative Agreement

Start Date: Sep 15, 2004 End Date: Sep 14, 2009

Objective: The objective of this cooperative research project with Dr. Maria Caramelli from the Italian BSE Reference Laboratory in Turin, Italy, is to conduct comparative studies with the U.S. bovine spongiform encephalopathy (BSE) isolate and the atypical BSE isolates identified in Italy. The studies will cover the following areas: 1. Evaluation of present diagnostics tools used in the U.S. for the detection of atypical BSE cases. 2. Molecular comparison of the U.S. BSE isolate and other typical BSE isolates with atypical BSE cases. 3. Studies on transmissibility and tissue distribution of atypical BSE isolates in cattle and other species.

Approach: This project will be done as a Specific Cooperative Agreement with the Italian BSE Reference Laboratory, Istituto Zooprofilattico Sperimentale del Piemonte, in Turin, Italy. It is essential for the U.S. BSE surveillance program to analyze the effectiveness of the U.S diagnostic tools for detection of atypical cases of BSE. Molecular comparisons of the U.S. BSE isolate with atypical BSE isolates will provide further characterization of the U.S. BSE isolate. Transmission studies are already underway using brain homogenates from atypical BSE cases into mice, cattle and sheep. It will be critical to see whether the atypical BSE isolates behave similarly to typical BSE isolates in terms of transmissibility and disease pathogenesis. If transmission occurs, tissue distribution comparisons will be made between cattle infected with the atypical BSE isolate and the U.S. BSE isolate. Differences in tissue distribution could require new regulations regarding specific risk material (SRM) removal.


http://www.ars.usda.gov/research/projects/projects.htm?ACCN_NO=408490




???$$$???



2009 UPDATE ON ALABAMA AND TEXAS MAD COWS 2005 and 2006


http://bse-atypical.blogspot.com/2006/08/bse-atypical-texas-and-alabama-update.html





P.4.25

Human susceptibility to atypical scrapie

Chris Plinston, Rona Barron, Nora Hunter The Roslin Institute and R(D)SVS, University of Edinburgh, UK

Background: Isolates of classical sheep scrapie are thought to pose little risk to humans as there have been no documented links between presence of sheep scrapie and the development of human TSE disease. However, the link between BSE and the development of vCJD in humans proves that a risk does exist from ruminant TSE disease, and therefore all new ruminant TSEs may potentially be transmissible to humans. Due to increased sensitivity of TSE diagnostic assay systems, a new TSE of sheep termed ‘atypical scrapie’ has been identified. This disease has been difficult to identify, and is found mainly in sheep which are previously thought to have a genetic makeup that made them resistant to scrapie. It is unclear whether this is a new TSE of sheep, an old disease which has only been identified through increased surveillance, or if it represents the phenotype of classical scrapie in so called ‘resistant’ sheep PrP genotypes.

Objectives: The objective of the study is to assess relative transmissibility of atypical scrapie isolates to humans and the associated risk to the population.

Methods: In order to determine whether atypical scrapie poses a risk to human health we have transmitted isolates from three different sheep PrP genotypes to our gene targeted transgenic mice which express human PrP with the M129V polymorphism known to be important in human susceptibility to disease. Mice of all three PrP genotypes have been inoculated intracerebrally with atypical scrapie isolates.

Discussion: In order to prevent the emergence of a new human TSE, we need to be able to assess the risk to humans from new emerging TSEs in livestock. The study of atypical scrapie infection in these transgenic lines could therefore provide important information on the host range and disease characteristics associated with such isolates. Preventative measures could then be put in place before this disease gives rise to another human disease variant and an underlying level of infection in the population.

P.5.21

Parallels between different forms of sheep scrapie and types of Creutzfeldt-Jakob disease (CJD)

Wiebke M. Wemheuer1, Sylvie L. Benestad2, Arne Wrede1, Wilhelm E. Wemheuer3, Tatjana Pfander1, Bjørn Bratberg2, Bertram Brenig3,Walter J. Schulz-Schaeffer1 1University Medical Center Goettingen, Germany; 2Institute of Veterinary Medicine Oslo, Norway; 3Institute of Veterinary Medicine Goettingen, Germany

Background: Scrapie in sheep and goats is often regarded as the archetype of prion diseases. In 1998, a new form of scrapie – atypical/Nor98 scrapie – was described that differed from classical scrapie in terms of epidemiology, Western blot profile, the distribution of pathological prion protein (PrPSc) in the body and its stability against proteinase K. In a similar way, distinct disease types exist in sporadic Creutzfeldt-Jakob disease (CJD). They differ with regard to their clinical outcome, Western blot profile and PrPSc deposition pattern in the central nervous system (CNS).

Objectives: The comparison of PrPSc deposits in sheep scrapie and human sporadic CJD. Methods: Tissues of the CNS of sheep with classical scrapie, sheep with atypical/Nor98 scrapie and 20 patients with sporadic CJD were examined using the sensitive Paraffin Embedded Tissue (PET) blot method. The results were compared with those obtained by immunohistochemistry. With the objective of gaining information on the protein conformation, the PrPSc of classical and atypical/Nor98 sheep scrapie and sporadic CJD was tested for its stability against denaturation with guanidine hydrochloride (GdnHCl) using a Membrane Adsorption Assay.

Results: The PrPSc of atypical/Nor98 scrapie cases and of CJD prion type 1 patients exhibits a mainly reticular/synaptic deposition pattern in the brain and is relatively sensitive to denaturation with GdnHCl. In contrast classical scrapie cases and CJD prion type 2 patients have a more complex PrPSc deposition pattern in common that consists of larger PrPSc aggregates and the PrPSc itself is comparatively stable against denaturation.

Discussion: The similarity between CJD types and scrapie types indicates that at least two comparable forms of the misfolded prion protein exist beyond species barriers and can elicit prion diseases. It seems therefore reasonable to classify classical and atypical/Nor98 scrapie – in analogy to the existing CJD types – as different scrapie types.

P.4.31

Prion infectivity in milk from ARQ/ARQ sheep experimentally infected with Scrapie and MAEDI-VISNA virus

Ciriaco Ligios1, Maria Giovanna Cancedda1, Antonello Carta2, Cinzia Santucciu1 Caterina Maestrale1, Francesca Demontis1, Sonia Attene1, Maria Giovanna Tilocca1, Cristiana Patta1, Massimo Basagni5, Paola Melis1, James C. De- Martini3, Christina Sigurdson4 1Istituto Zooprofilattico Sperimentale della Sardegna, Italy; 2Research Unit: Genetics and Biotechnology, DIRPA, AGRIS Sardinia, Italy; 3Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO, USA; 4Department of Pathology, School of Medicine, University of California San Diego, USA; 5Prion Diagnostica Rho, Italy

Background: Scrapie in sheep is characterized by the deposition of misfolded and aggregated prion protein (PrPSc) in the central nervous system (CNS) and within the lymphoreticular system (LRS). PrPSc was shown to accumulate in organs beyond the CNS and the LRS when lymphofollicular or granulomatous inflammation was also present. Objectives: Our aim was to determine whether ectopic PrPSc accumulation in the inflamed mammary gland of sheep with scrapie results in infectious prion secretion into the milk.

Methods: We fed approximately 1.1 - 2.1 L of milk from sheep with lymphofollicular mastitis and clinical scrapie to each of 8 ARQ/ARQ lambs derived from scrapie-free flocks. The milk donor sheep had been previously inoculated with Maedi-Visna virus (MVV) intratracheally and intravenously and scrapie brain homogenate orally. In addition, 3 ARQ/ARQ lambs were fed approximately 1.4 – 1.7 L of milk from ARQ/ARQ sheep that had been experimentally infected with only scrapie. Additional control ARQ/ARQ lambs were inoculated with scrapie brain homogenate only, or with milk from uninfected sheep.

Results: Two lambs which had received milk from sheep with mastitis and scrapie developed clinical signs of scrapie at 677 and 745 days post-inoculation. One additional clinically healthy lamb from this group, which was sacrificed for a cause unrelated to scrapie, was found to have PrPSc in brain and tonsil. The control lambs and those which received milk from sheep affected only with scrapie are, to date, clinically healthy.

Discussion: This is the first evidence of clinical scrapie in sheep fed milk from scrapie sick sheep. The experiment is ongoing, however these preliminary results indicate that milk and/or colostrum from ARQ/ARQ sheep with clinical scrapie and lymphofollicular mastitis could contribute to scrapie transmission.

P.4.50

Successful oral transmission of classical scrapie to ARR/ARQ sheep

Sarah Jo Moore1, Hugh Simmons1, Timm Konold1, Glenda Dexter1, Steve Ryder2 1Veterinary Laboratories Agency, 2Home Office Animals (Scientific Procedures) Inspectorate

Background: Scrapie susceptibility in sheep is strongly influenced by allelic variation in the gene which encodes the prion protein. As part of the National Scrapie Plan (NSP) for Great Britain ram genotyping and selective breeding has been used to increase the number of sheep in the national flock that are genetically resistant to classical scrapie. According to the NSP, ARR/ARQ sheep are considered ‘genetically resistant’ to scrapie, although four field cases have been detected since 2002.

Objective: To investigate the susceptibility and pathogenesis of classical scrapie in ARR/ARQ sheep.

Methods: TSE-free lambs were dosed orally with 5g of pooled brain from scrapie clinical suspects. Timed-culls were performed at 12 and 24 months post-inoculation (mpi) then six monthly thereafter. All sheep underwent a detailed clinical examination before culling. At post-mortem 41 tissues were sampled from all major body systems. Detection of disease-associated prion protein (PrPd) in central nervous system tissues was done by immunohistochemistry (IHC), Western blot and Bio-rad elisa. All other tissues were examined by IHC only.

Results: PrPd was first detected in the LRS at 24 mpi, in the central nervous system (CNS) at 36 mpi, and in the peripheral nervous system (PNS) at 66 mpi. Throughout the time course PrPd accumulation in LRS tissues was more restricted and less severe than in CNS and PNS tissues. The first confirmed clinical case occurred at 72 mpi.

Discussion: We have shown that ARR/ARQ sheep can be infected with classical scrapie via the oral route.The pathogenesis of scrapie in ARR/ARQ sheep appears to be different to that in sheep of susceptible genotypes. While VRQ/VRQ clinical suspects have extensive LRS involvement only one LRS tissue was positive in the ARR/ARQ clinical suspect. This could suggest that infectivity had travelled to the CNS via a nongastrointestinal route. ARR/ARQ sheep may act as ‘silent carriers’ of disease. However, PrPd accumulation in the gut-associated lymphoid tissues was mild and restricted so non-faecal routes may be more important in lateral transmission from this genotype. The tissue distribution of PrPd accumulation in this study suggest that currently available ‘live tests’ for preclinical diagnosis – third eyelid and/or rectoanal mucosa-associated lymphoid tissue (RAMALT) biopsy – may be unrewarding in sheep of this genotype.


http://www.prion2009.com/sites/default/files/Prion2009_Book_of_Abstracts.pdf




Wednesday, July 1, 2009

Nor98 scrapie identified in the United States J Vet Diagn Invest 21:454-463 (2009)


http://nor-98.blogspot.com/2009/07/nor98-scrapie-identified-in-united.html




P03.141

Aspects of the Cerebellar Neuropathology in Nor98

Gavier-Widén, D1; Benestad, SL2; Ottander, L1; Westergren, E1 1National Veterinary Insitute, Sweden; 2National Veterinary Institute,

Norway Nor98 is a prion disease of old sheep and goats. This atypical form of scrapie was first described in Norway in 1998. Several features of Nor98 were shown to be different from classical scrapie including the distribution of disease associated prion protein (PrPd) accumulation in the brain. The cerebellum is generally the most affected brain area in Nor98. The study here presented aimed at adding information on the neuropathology in the cerebellum of Nor98 naturally affected sheep of various genotypes in Sweden and Norway. A panel of histochemical and immunohistochemical (IHC) stainings such as IHC for PrPd, synaptophysin, glial fibrillary acidic protein, amyloid, and cell markers for phagocytic cells were conducted. The type of histological lesions and tissue reactions were evaluated. The types of PrPd deposition were characterized. The cerebellar cortex was regularly affected, even though there was a variation in the severity of the lesions from case to case. Neuropil vacuolation was more marked in the molecular layer, but affected also the granular cell layer. There was a loss of granule cells. Punctate deposition of PrPd was characteristic. It was morphologically and in distribution identical with that of synaptophysin, suggesting that PrPd accumulates in the synaptic structures. PrPd was also observed in the granule cell layer and in the white matter. The pathology features of Nor98 in the cerebellum of the affected sheep showed similarities with those of sporadic Creutzfeldt-Jakob disease in humans.

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


http://www.prion2007.com/pdf/Prion%20Book%20of%20Abstracts.pdf




PR-26

NOR98 SHOWS MOLECULAR FEATURES REMINISCENT OF GSS

R. Nonno1, E. Esposito1, G. Vaccari1, E. Bandino2, M. Conte1, B. Chiappini1, S. Marcon1, M. Di Bari1, S.L. Benestad3, U. Agrimi1 1 Istituto Superiore di Sanità, Department of Food Safety and Veterinary Public Health, Rome, Italy (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000164/!x-usc:mailto:romolo.nonno@iss.it); 2 Istituto Zooprofilattico della Sardegna, Sassari, Italy; 3 National Veterinary Institute, Department of Pathology, Oslo, Norway

Molecular variants of PrPSc are being increasingly investigated in sheep scrapie and are generally referred to as "atypical" scrapie, as opposed to "classical scrapie". Among the atypical group, Nor98 seems to be the best identified. We studied the molecular properties of Italian and Norwegian Nor98 samples by WB analysis of brain homogenates, either untreated, digested with different concentrations of proteinase K, or subjected to enzymatic deglycosylation. The identity of PrP fragments was inferred by means of antibodies spanning the full PrP sequence. We found that undigested brain homogenates contain a Nor98-specific PrP fragment migrating at 11 kDa (PrP11), truncated at both the C-terminus and the N-terminus, and not N-glycosylated. After mild PK digestion, Nor98 displayed full-length PrP (FL-PrP) and N-glycosylated C-terminal fragments (CTF), along with increased levels of PrP11. Proteinase K digestion curves (0,006-6,4 mg/ml) showed that FL-PrP and CTF are mainly digested above 0,01 mg/ml, while PrP11 is not entirely digested even at the highest concentrations, similarly to PrP27-30 associated with classical scrapie. Above 0,2 mg/ml PK, most Nor98 samples showed only PrP11 and a fragment of 17 kDa with the same properties of PrP11, that was tentatively identified as a dimer of PrP11. Detergent solubility studies showed that PrP11 is insoluble in 2% sodium laurylsorcosine and is mainly produced from detergentsoluble, full-length PrPSc. Furthermore, among Italian scrapie isolates, we found that a sample with molecular and pathological properties consistent with Nor98 showed plaque-like deposits of PrPSc in the thalamus when the brain was analysed by PrPSc immunohistochemistry. Taken together, our results show that the distinctive pathological feature of Nor98 is a PrP fragment spanning amino acids ~ 90-155. This fragment is produced by successive N-terminal and C-terminal cleavages from a full-length and largely detergent-soluble PrPSc, is produced in vivo and is extremely resistant to PK digestion.

*** Intriguingly, these conclusions suggest that some pathological features of Nor98 are reminiscent of Gerstmann-Sträussler-Scheinker disease.

119


http://www.neuroprion.com/pdf_docs/conferences/prion2006/abstract_book.pdf




A newly identified type of scrapie agent can naturally infect sheep with resistant PrP genotypes

Annick Le Dur*,?, Vincent Béringue*,?, Olivier Andréoletti?, Fabienne Reine*, Thanh Lan Laï*, Thierry Baron§, Bjørn Bratberg¶, Jean-Luc Vilotte?, Pierre Sarradin**, Sylvie L. Benestad¶, and Hubert Laude*,?? +Author Affiliations

*Virologie Immunologie Moléculaires and ?Génétique Biochimique et Cytogénétique, Institut National de la Recherche Agronomique, 78350 Jouy-en-Josas, France; ?Unité Mixte de Recherche, Institut National de la Recherche Agronomique-Ecole Nationale Vétérinaire de Toulouse, Interactions Hôte Agent Pathogène, 31066 Toulouse, France; §Agence Française de Sécurité Sanitaire des Aliments, Unité Agents Transmissibles Non Conventionnels, 69364 Lyon, France; **Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, 37380 Nouzilly, France; and ¶Department of Pathology, National Veterinary Institute, 0033 Oslo, Norway

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

Abstract Scrapie in small ruminants belongs to transmissible spongiform encephalopathies (TSEs), or prion diseases, a family of fatal neurodegenerative disorders that affect humans and animals and can transmit within and between species by ingestion or inoculation. Conversion of the host-encoded prion protein (PrP), normal cellular PrP (PrPc), into a misfolded form, abnormal PrP (PrPSc), plays a key role in TSE transmission and pathogenesis. The intensified surveillance of scrapie in the European Union, together with the improvement of PrPSc detection techniques, has led to the discovery of a growing number of so-called atypical scrapie cases. These include clinical Nor98 cases first identified in Norwegian sheep on the basis of unusual pathological and PrPSc molecular features and "cases" that produced discordant responses in the rapid tests currently applied to the large-scale random screening of slaughtered or fallen animals. Worryingly, a substantial proportion of such cases involved sheep with PrP genotypes known until now to confer natural resistance to conventional scrapie. Here we report that both Nor98 and discordant cases, including three sheep homozygous for the resistant PrPARR allele (A136R154R171), efficiently transmitted the disease to transgenic mice expressing ovine PrP, and that they shared unique biological and biochemical features upon propagation in mice. These observations support the view that a truly infectious TSE agent, unrecognized until recently, infects sheep and goat flocks and may have important implications in terms of scrapie control and public health.


http://www.pnas.org/content/102/44/16031.abstract




Monday, December 1, 2008

When Atypical Scrapie cross species barriers

Authors

Andreoletti O., Herva M. H., Cassard H., Espinosa J. C., Lacroux C., Simon S., Padilla D., Benestad S. L., Lantier F., Schelcher F., Grassi J., Torres, J. M., UMR INRA ENVT 1225, Ecole Nationale Veterinaire de Toulouse.France; ICISA-INlA, Madrid, Spain; CEA, IBiTec-5, DSV, CEA/Saclay, Gif sur Yvette cedex, France; National Veterinary Institute, Postboks 750 Sentrum, 0106 Oslo, Norway, INRA IASP, Centre INRA de Tours, 3738O Nouzilly, France.

Content

Atypical scrapie is a TSE occurring in small ruminants and harbouring peculiar clinical, epidemiological and biochemical properties. Currently this form of disease is identified in a large number of countries. In this study we report the transmission of an atypical scrapie isolate through different species barriers as modeled by transgenic mice (Tg) expressing different species PRP sequence.

The donor isolate was collected in 1995 in a French commercial sheep flock. inoculation into AHQ/AHQ sheep induced a disease which had all neuro-pathological and biochemical characteristics of atypical scrapie. Transmitted into Transgenic mice expressing either ovine or PrPc, the isolate retained all the described characteristics of atypical scrapie.

Surprisingly the TSE agent characteristics were dramatically different v/hen passaged into Tg bovine mice. The recovered TSE agent had biological and biochemical characteristics similar to those of atypical BSE L in the same mouse model. Moreover, whereas no other TSE agent than BSE were shown to transmit into Tg porcine mice, atypical scrapie was able to develop into this model, albeit with low attack rate on first passage.

Furthermore, after adaptation in the porcine mouse model this prion showed similar biological and biochemical characteristics than BSE adapted to this porcine mouse model. Altogether these data indicate.

(i) the unsuspected potential abilities of atypical scrapie to cross species barriers

(ii) the possible capacity of this agent to acquire new characteristics when crossing species barrier

These findings raise some interrogation on the concept of TSE strain and on the origin of the diversity of the TSE agents and could have consequences on field TSE control measures.


http://www.neuroprion.org/resources/pdf_docs/conferences/prion2008/abstract-book-prion2008.pdf




NOR-98 ATYPICAL SCRAPIE 5 cases documented in USA in 5 different states USA 2007


http://nor-98.blogspot.com/2008/04/seac-spongiform-encephalopathy-advisory.html




Tuesday, June 3, 2008 SCRAPIE USA UPDATE JUNE 2008 NOR-98 REPORTED PA


http://nor-98.blogspot.com/2008/06/scrapie-usa-update-june-2008-nor-98.html




Monday, September 1, 2008

RE-FOIA OF DECLARATION OF EXTRAORDINARY EMERGENCY BECAUSE OF AN ATYPICAL T.S.E. (PRION DISEASE) OF FOREIGN ORIGIN IN THE UNITED STATES [No. 00-072-1] September 1, 2008


http://foiamadsheepmadrivervalley.blogspot.com/2008/09/re-foia-of-declaration-of-extraordinary.html




http://nor-98.blogspot.com/




http://scrapie-usa.blogspot.com/




O.11.2

Transmission of bovine-passaged TME prion strain to macaque

Emmanuel Comoy1, Juergen Richt2, Valérie Durand1, Sophie Freire1, Evelyne Correia1, Amir Hamir2, Marie- Madeleine Ruchoux1, Paul Brown1, Jean-Philippe Deslys1 1Atomic Energy Commission, France; 2National Animal Disease Center, USA

Background: The origin of Transmissible Mink Encephalopathy (TME) remains controversial, with historical evidence for either scrapie or BSE as the source of separate outbreaks. The case for BSE is supported by the experimental transmission of BSE from cattle to mink, whereas scrapie failed to transmit from sheep to mink. Transmission of TME from mink to cynomolgus macaque is inefficient, suggesting a low risk of TSE to human health. Because only typical and atypical BSE prion strains have been shown to be easily transmissible from non-primate to primate species, we have investigated transmissibility to monkeys of a cattle-passaged strain of TME.

Objectives: To compare the transmissibility of cattle-passaged TME prions to the transmissibility of other cattle-passaged prions.

Methods: Monkeys (cynomolgus macaques) were intra-cerebrally infected with classical BSE, atypical BSE strains (BASE and BSE H), and a cattle-passaged TME strain. Animals were regularly monitored for clinical signs, and extensive biochemical and immunohistochemical studies were performed on lymphoid and neural tissues of animals that have already died.

Results and discussion: The animal infected with the cattlepassaged TME strain developed neurological clinical signs after a very short incubation period of 20 months, with a clinical picture that is clearly different from that of BSE/vCJD-infected animals, but similar to that of BASE (the animal is still alive at the time of this writing but post-mortem histopathological and immunohistochemical analyses will provide a more complete characterization of the disease). This new transmission reinforces the notion of human vulnerability to prion diseases passaged through cattle, perhaps due to a low species barrier.


http://www.prion2009.com/sites/default/files/Prion2009_Book_of_Abstracts.pdf




Saturday, December 01, 2007

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


http://transmissible-mink-encephalopathy.blogspot.com/2007/12/phenotypic-similarity-of-transmissible.html




Tuesday, July 21, 2009

Transmissible mink encephalopathy - review of the etiology Folia Neuropathologica 2/2009


http://transmissible-mink-encephalopathy.blogspot.com/2009/07/transmissible-mink-encephalopathy.html



http://transmissible-mink-encephalopathy.blogspot.com/




O.4.5

Recent results on the transmission, detection, and pathogenesis of chronic wasting disease

Edward A. Hoover1, Nicholas J. Haley1, Candace K. Mathiason1, Nathanial D. Denkers1, Davis M. Seelig1, and Glenn C. Telling2 1Colorado State University, USA; 2University of Kentucky, Lexington, USA

Background: Chronic wasting disease (CWD) of cervids is distinguished by its high level of transmissibility. We have previously shown that body fluids and excretions contain infectious CWD prions. The precise means by which these prions may access, traffick and cause disease in cervids (or other species) remains to be elucidated. Here we present current results of studies employing cervid and cervidized mouse bioassays and serial protein misfolding cyclic amplification (sPMCA) to address these questions.

Objectives: We sought to determine: (1) which components of blood and saliva carry prion infectivity; (2) whether long term, very low level CWD infection undetectable by conventional assays may exist in cervids; (3) whether CWD can be transmitted via aerosol or minor oral lesions; and (4) potential alternate pathways of CWD prion entry and dissemination exist in vivo.

Methods: The studies described utilized cervid and cervid-PrPexpressing transgenic mouse bioassays, serial protein misfolding cyclic amplification (sPMCA), and high resolution immunostaining.

Results: We present data to demonstrate: (1) localization of infectious CWD prions chiefly to the circulating CD21-expressing B/DC cell fractions of blood; (2) very low level subclinical CWD infection in cervids detectable by sPMCA and bioassay; (3) CWD transmission by aerosol and minor oral epithelial lesions; and (4) evidence of villous autonomic neural uptake and dissemination of PrPCWD.

Discussion: These findings contribute insights into CWD prion transmission, trafficking, and dissemination. The results also help direct efforts toward ante-mortem detection of CWD in cervids and raise interesting questions regarding duration of sub-clinical prion infection in cervids or other species.

O.11.1

Prions in feces of asymptomatic deer

Gültekin Tamgüney1,2, Michael W. Miller3, Lisa L. Wolfe3, Tracey M. Sirochman3, David V. Glidden4, Christina Palmer 1, Azucena Lemus5, Stephen J. DeArmond5, Stanley B. Prusiner1,2 1Institute for Neurodegenerative Diseases, University of California, San Francisco, USA; 2Department of Neurology, University of California, San Francisco, USA; 3Colorado Division of Wildlife, Wildlife Research Center, Fort Collins, USA; 4Department of Epidemiology and Biostatistics, University of California, San Francisco, USA; 5Department of Pathology, University of California, San Francisco, USA

Background: Chronic wasting disease (CWD) of several species in the deer family and scrapie of sheep are infectious prion diseases that are transmitted naturally within affected host populations. Even though several potential sources of infectivity have been identified in secretions and excretions from symptomatic animals, the biological importance of these sources in sustaining epidemics remains unclear.

Objective/Methods: Feces from mule deer (Odocoileus hemionus) were periodically collected before and after oral inoculation with CWD prions until the deer developed clinical signs of CWD. Fecal samples were irradiated and intracerebrally inoculated into transgenic mice overexpressing cervid PrP. Results: We report that asymptomatic CWD-infected mule deer excrete CWD prions in their feces long before they develop clinical signs of prion disease. Intracerebral inoculation of irradiated deer feces into transgenic mice overexpressing cervid PrP revealed infectivity in 14 of 15 fecal samples collected from 5 deer at 7–11 months before the onset of neurological disease. Even though prion concentrations in deer feces were much lower than those in brain tissue from the same deer collected at the disease terminus, the estimated total infectious dose excreted in feces by an infected deer over the disease course may approximate the total contained in brain tissue.

Discussion: Fecal prion excretion over long periods of time by infected deer provides a likely natural mechanism that may explain the high incidence and efficient horizontal transmission of CWD within deer herds, as well as prion transmission among susceptible cervid species.

P.4.11

Detection of subclinical CWD infection in conventional test-negative deer long after oral exposure to urine and feces from CWD+ deer

Nicholas Haley1, Candace Mathiason1, Mark Zabel1, Glenn Telling2, Edward Hoover1 1Colorado State University, USA; 2University of Kentucky, USA

Background: Chronic wasting disease (CWD) of cervids is distinguished by its high level of transmissibility, wherein bodily fluids and excretions are thought to play an important role. Using cervid bioassay and established CWD detection methods, we have previously identified infectious prions in saliva and blood but not urine or feces of CWD+ donors. More recently, we were able to identify very low concentrations of CWD prions in urine of deer by cervid PrP transgenic (Tg[Cer- PrP]) mouse bioassay and serial protein misfolding cyclic amplification (sPMCA).

Objectives: In these experiments, we sought to investigate whether deer previously exposed orally to urine and feces from CWD+ deer, while conventional test-negative, may actually be harboring very low level CWD infection, not evident in the 19 month observation period in the previous studies. Methods: Brain and lymph nodes from conventional test-negative deer were reanalyzed for CWD prions by sPMCA and cervid transgenic mouse bioassay in parallel with appropriate tissue-matched positive and negative controls.

Results: PrPres was detected in tissues of exposed deer by both sPMCA and Tg[CerPrP] mouse bioassay; each assay revealed very low levels of CWD prions previously undetectable by western blot, ELISA, or IHC. D

iscussion: The finding of subclinical infection in deer orally exposed to urine and feces (1) suggests that a prolonged subclinical state can exist such that observation periods in excess of two years may be needed to detect CWD infection and (2) illustrates the sensitive and specific application of sPMCA in the diagnosis of low level prion infection.

P.4.27

Minor oral lesions facilitate CWD infection

Nathaniel Denkers1, Glenn Telling2, Edward Hoover1 1Colorado State University, USA; 2University of Kentucky, USA

Background: While the exact mechanisms of chronic wasting disease (CWD) prion transmission, entry, and trafficking remain incompletely elucidated, transmission by exposure of the oral and/or nasal mucous membranes seems certain. As part of foraging, cervids likely experience minor lesions in the oral mucous membranes; these could have impact on susceptibility to prion entry and subsequent infection.

Objectives: To explore this potential co-factor, we used cervid PrP transgenic mice to assess whether or not micro-abrasions to the tongue may enhance susceptibility to oral CWD infection.

Methods: Two sets of FVB mice transgenically expressing the normal cervid PrPC protein [Tg(cerPrP) mice], with or without abrasions on the lingual mucosa, were inoculated orally with 10ìl of a 10% w/v brain homogenate from either CWD-positive or negative deer. Abrasions were created by lightly scratching the dorsal lingual epithelium with a 30g needle. Cohorts were sacrificed at 1, 2, 12, 52, 78, and 104 weeks post inoculation (pi) or when signs of neurologic disease were observed. Tongue, lymphoid tissue, and the brain were assessed by western blotting and immunohistochemistry to detect the CWD abnormal prion protein (PrPCWD).

Results: Between 296 and 430 dpi, 8 of the 9 CWD-inoculated mice with lingual lesions developed clinical signs of neurologic dysfunction mandating euthanasia. The brains of all 8 mice were positive by western blot and immunohistochemistry for PrPCWD. Conversely, all mice without oral lesions remain asymptomatic at >450 dpi. No evidence of PrPCWD was detected in any Tg(cerPrP) mice examined at any of the preterminal time points.

Discussion: Micro-abrasions to the lingual surface substantially facilitate CWD transmission, suggesting a co-factor that may be significant in foraging cervids or other species. Earlier post-inoculation sampling intervals (1 and 4 hours) are in progress in an attempt to determine when and where PrPCWD might be detectable after oral mucosal exposure.

P.4.26

Aerosol and intranasal transmission of CWD

Nathaniel Denkers1, Glenn Telling2, Edward Hoover1 1Colorado State University, USA; 2University of Kentucky, USA

Background: Little is known regarding the potential risk posed by aerosolized prions. Chronic wasting disease (CWD) prions are present in saliva and urine of infected animals and it is clearly established that CWD is transmitted horizontally, almost surely by mucosal exposure. However, the potential transmissibility of CWD by aerosol or nasal routes is not known.

Objectives: The present study was therefore designed to determine whether CWD prions are transmissible by these routes of exposure using the cervid PrP transgenic mouse model of CWD infection.

Methods: FVB mice transgenically expressing the normal cervid PrPC protein [Tg(cerPrP) mice] were exposed to CWD prions by either nose-only exposure to an aerosol generated by nebulizing 0.5 ml of a 5% w/v CWD+ brain homogenate or 10ƒÊl of a 10% w/v CWD+ brain homogenate by dropwise instillation into the nostrils. Mice were monitored for signs of clinical disease for up to 755 days post inoculation (dpi). Nasal mucosa, vomeronasal organ, lymphoid tissue, and the brain were assessed for PrPCWD by western blotting and immunohistochemistry.

Results: Six of 7 aerosol-exposed Tg(cerPrP) mice developed clinical signs of neurologic dysfunction between 411 and 749 dpi mandating euthanasia. In all symptomatic mice CWD infection was confirmed by histopathologic lesions and detection of PrPCWD within the brain. Two of 9 IN-inoculated Tg(cerPrP) mice also developed TSE between 417 and 755 dpi, again confirmed by PrPCWD detection within the brain. No evidence of PrPCWD was detected in any Tg(cerPrP) mice examined at any of the pre-terminal time points.

Discussion: CWD is transmissible by aerosol as well as intranasal exposurePpotentially implicating exposure via the respiratory system in CWD and potentially other prion diseases. Studies examining very early post-inoculation sampling intervals (1 and 4 hours) are in progress in an attempt to determine initial prion targeting and entry portals.

P.10.15

Adaptation of chronic wasting disease (CWD) into hamsters: evidence of a novel strain of CWD

Chad Johnson1, Debbie McKenzie2 1University of Wisconsin-Madison, USA, 2University of Alberta, Canada

Background: Prion strains are well-characterized for scrapie and BSE. Little is known about the potential for strains in chronic wasting disease (CWD). Different CWD strains could have different patterns of shedding infectious agent as well as differential detectiblility and interspecies transmissibility.

Objectives: We identified prion protein variants in whitetailed deer populations and demonstrated that Prnp genotype affects the susceptibility/disease progression of white-tailed deer to CWD. Cervid prion protein variants raise the likelihood of distinct CWD strains. We hypothesize that the prion protein variability results in different PrPCWD conformers producing different CWD strains upon interspecies transmission.

Methods: Hamsters were intracerebrally inoculated with brain homogenate or phosphotungstate concentrated preparations from CWD positive hunter-harvested (Wisconsin CWD endemic area) and from experimentally infected deer with known Prnp genotypes.

Results: Primary passage of concentrated CWD agent resulted in clinical disease at approximately 1 year post-infection. Subclinical infection was established with the other primary passages based on the detection of PrPCWD in the brains of hamsters and the successful disease transmission upon second passage. Second and third passage resulted in a decrease in incubation period to approximately 280 days. Inocula from deer with wt/wt and wt/G96S Prnp genotypes did not result in any differences in incubation period or clinical symptoms.

Discussion: Inocula from deer with the wt/wt and wt/G96S prion proteins resulted in similar disease upon transmission to hamsters, likely due to the presence of the wt prion protein. Comparison of the incubation period and clinical symptoms of the hamsters infected with inocula from Wisconsin whitetailed deer to transmission studies using different CWD inocula (Western white-tailed deer; elk and mule deer; Raymond et al. 2007) indicates that the CWD agent present in US Midwest is different from the strain(s) present in the endemic region of the western US.

O.9.2

Survival and limited spread of TSE infectivity after burial for one year

Robert Somerville, Karen Fernie, Allister Smith University of Edinburgh, UK

Background: Scrapie and Chronic Wasting Disease appear to spread via environmental routes, although there is little evidence that BSE or CJD do. Nevertheless there are concerns about reservoirs of BSE infection remaining in the environment after carcass burial or waste disposal.

Objectives: We are determining the survival and migration of TSE infectivity when buried for up to five years in two soil types; and either buried within bovine skulls, or as a point source bolus of TSE infected brain.

Methods: Two demonstration experiments have been set up. In one experiment boluses of TSE infected mouse brain (301V strain) have been buried in lysimeters containing either a sandy soil or a clay soil. Migration from the boluses is being assessed from soil cores taken from the lysimeters over time and assayed for TSE infectivity. In the other experiment ten bovine heads have been spiked with TSE infected mouse brain (301V strain) and buried in the two soil types. Two heads are exhumed annually and assessed for residual infectivity within and around them.

Results: After one year very small amounts of infectivity have been detected 25cm from the point source bolus in both soils. No infectivity was detected up to 9 months after burial. In the other experiment brain-like tissue was still apparent within the crania after one year. Large amounts of TSE infectivity were detected from samples of the intracranial contents, and also detected in very small amounts in soil samples in the soil immediately surrounding the heads.

Discussion: These data show that TSE infectivity can survive burial for up to a year but migrates very slowly in these soils. Results from future years will likely further illustrate the long term survival and migration properties of these infective agents. Risk assessments of TSE infectivity in the environment should take into account the likely long survival rate of foci of infectivity when large amounts of infected material have been buried.


http://www.prion2009.com/sites/default/files/Prion2009_Book_of_Abstracts.pdf




please note not only first passage, but also second passage of CWD transmission studies to cattle, and please see attack rate ;

Interpretive Summary: This study reports findings assessing susceptibility of cattle to infection following direct surgical inoculation of the transmissible spongiform encephalopathy (TSE), chronic wasting disease (CWD, from white tailed deer) into the brain of 14 cattle. Three-month-old calves were inoculated with the CWD agent from white tailed deer. Two non-inoculated calves served as controls. Within 26 months post inoculation, 12 inoculated animals had lost weight, revealed abnormal clinical signs, and were euthanatized. Laboratory tests revealed the presence of a unique pattern of the disease agent in tissues of these animals. These findings demonstrate that when CWD is directly inoculated into the brain of cattle, 86% of inoculated cattle develop clinical signs of the disease. ...

see full text ;

http://www.ars.usda.gov/research/publications/publications.htm?seq_no_115=194089



see also;

Vet Pathol 44:487-493 (2007) © 2007 American College of Veterinary Pathologists

Susceptibility of Cattle to First-passage Intracerebral Inoculation with Chronic Wasting Disease Agent from White-tailed Deer A. N. Hamir, J. M. Miller, R. A. Kunkle, S. M. Hall and J. A. Richt National Animal Disease Center, ARS, USDA, Ames, IA (ANH, JMM, RAK, JAR), Pathobiology Laboratory, National Veterinary Services Laboratories, Ames, IA (SMH)

Fourteen, 3-month-old calves were intracerebrally inoculated with the agent of chronic wasting disease (CWD) from white-tailed deer (CWDwtd) to compare the clinical signs and neuropathologic findings with those of certain other transmissible spongiform encephalopathies (TSE, prion diseases) that have been shown to be experimentally transmissible to cattle (sheep scrapie, CWD of mule deer [CWDmd], bovine spongiform encephalopathy [BSE], and transmissible mink encephalopathy). Two uninoculated calves served as controls. Within 26 months postinoculation (MPI), 12 inoculated calves had lost considerable weight and eventually became recumbent. Of the 12 inoculated calves, 11 (92%) developed clinical signs. Although spongiform encephalopathy (SE) was not observed, abnormal prion protein (PrPd) was detected by immunohistochemistry (IHC) and Western blot (WB) in central nervous system tissues. The absence of SE with presence of PrPd has also been observed when other TSE agents (scrapie and CWDmd) were similarly inoculated into cattle. The IHC and WB findings suggest that the diagnostic techniques currently used to confirm BSE would detect CWDwtd in cattle, should it occur naturally. Also, the absence of SE and a distinctive IHC pattern of CWDwtd and CWDmd in cattle suggests that it should be possible to distinguish these conditions from other TSEs that have been experimentally transmitted to cattle.

--------------------------------------------------------------------------------

Key words: Cattle; chronic wasting disease; prion diseases; prion protein immunohistochemistry; prion protein Western blot; spongiform encephalopathy.

Request reprints from Dr. A. N. Hamir, National Animal Disease Center, ARS, USDA, 2300 Dayton AvenuePO Box 70, Ames, IA 50010 (USA). E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000164/!x-usc:mailto:ahamir@nadc.ars.usda.gov

http://www.vetpathology.org/cgi/content/abstract/44/4/487



snip...

Discussion

This study used the intracerebral route of inoculation to provide the most efficient means of testing the absolute susceptibility of cattle to the agent of CWD from white-tailed deer and to provide a comparison with similarly designed previous studies.2-5,7 Such studies do not inform on susceptibility by natural routes of transmission but, if negative, can provide evidence supporting the existence of a considerable species barrier. In instances of successful transmission they also provide an understanding of the descriptive pathology that might characterize the natural disease in the recipient species.

CWD, like all other TSEs, is characterized by a long incubation period, which in deer is seldom less than 18 months.15,16 In an experimental study of cattle inoculated intracerebrally with CWD from mule deer (first passage; CWDmd), PrPd was demonstrated in only 5 of 13 cattle (38%), after incubation periods that ranged from 23 to 63 MPI.4,5 In contrast, 12 of 14 inoculated cattle in the present study were positive for PrPd (86% attack rate) within 26 MPI (21.5 ± 0.5 MPI). The increased incidence and shorter incubation periods indicate that the CWDwtd inoculum was more pathogenic in cattle than the CWDmd inoculum on first passage.4,5 However, there may be several reasons for these differences, such as genetic variability of the recipient hosts, the differences in PrP genotype of pooled brain material in the inoculum, and the infectivity titer of the inocula used in the CWD transmission studies of cattle.4,5 Regarding the latter, in a separate study of intracerebral CWD inoculation of white-tailed deer, we found both sources of CWD (from mule deer and white-tailed deer) successfully transmitted to each of two groups of white-tailed deer within two years, and there was no significant difference in incubation times and lesions between the groups (Kunkle et al., unpublished data).

In cervids, clinical CWD is characterized by emaciation, changes in behavior, and excessive salivation.15,16 Although the latter was not observed in the CWDwtd-inoculated cattle, 12 of 14 inoculated calves showed anorexia and considerable weight loss, and the majority of these cattle also showed intermittent abnormal CNS signs. However, despite the advanced clinical signs in the affected cattle, none of the animals had histopathologic changes of SE, but all were positive for PrPd by IHC and WB. The absence of morphologic lesions in cattle with cross-species transmitted TSEs has been documented previously with cattle inoculated intracerebrally with the sheep scrapie agent.3 In that study, 100% of cattle succumbed to the disease between 14 and 18 months after inoculation with a US strain of the scrapie agent derived from sheep. None of the cattle showed microscopic lesions, and all were positive for PrPd. A similar lack of morphologic changes was observed in cattle inoculated with CWDmd, even after the second passage.4,5,8

The predominant characteristics of IHC reactivity reported previously for cattle inoculated with mule deer CWD4,5,7 were also observed in the diseased cattle that died after inoculation with CWD from white tailed-deer. The PrPd accumulations were primarily multifocal and presumably glial cell-associated, with no evidence of intraneuronal or perineuronal labeling. The only difference noted between calves inoculated with CWDmd and CWDwtd was that the latter seemed to show more extensive labeling, both in amount and anatomic distribution within the CNS. In addition, labeling of white matter was more prominent in the CWDwtd-inoculated calves.

In this and in the earlier study of CWDmd in cattle,4,5 IHC labeling differed from that seen in cattle with bovine spongiform encephalopathy (BSE) or experimental transmissible mink encephalopathy (TME).6 In both (BSE and TME) the IHC labeling was similar and was characterized by widespread diffuse (usually perineuronal) labeling of grey matter neuropil, with labeled particles that were not cell-associated except occasionally at neuronal cell membranes.5,14 The IHC pattern in bovine CWD also contrasts markedly with that seen in scrapie-inoculated cattle, in which intracytoplasmic labeling of neurons was a prominent feature.2,3

Compared with experimental TME in cattle,7 the experimental bovine CWD in this study was associated with less extensive IHC labeling in neural tissues other than brain and spinal cord. Whereas the retina was positive in all cattle inoculated with TME, none of the CWD-infected cattle in this experiment had any retinal labeling. Similarly, in the present study there was no labeling in the pituitary gland, a tissue that in some TME-infected cattle is positive.7 The different affinities of tissues may reflect different abilities of these agents to amplify in these tissues. However, given sample sizes and uncontrolled experimental variables, other features, including host genetics, titers of inocula, and stage of disease cannot be excluded.

IHC labeling for PrPd was not observed in striated muscles (heart, tongue, masseter, diaphragm) of the experimental calves. This observation is in accordance with our previous findings9 in which striated muscle from 20 animals (cattle, sheep, elk, and raccoons) was examined for PrPd by IHC. In these animals, all of which had developed a TSE after experimental inoculation, PrPd was found by IHC examination in the CNS, but not in striated muscle. However, recent investigations with an enriched WB technique12 have enabled us to detect PrPd in the tongues of some sheep and elk that were experimentally inoculated with scrapie and CWD, respectively (Bessen et al., unpublished data).

When brainstems of CWD-infected cattle were analyzed by WB for the presence of PrPd, 12 of 14 samples were found to be positive for PrPd (Table 1; Fig. 3), showing a clear reaction with the three protease resistant polypeptide isoforms (di-, mono-, and unglycosylated). Interestingly, No. 3, which was found by IHC to be negative in the obex area, but positive in midbrain region (Table 2), was found positive by WB (Fig. 3). This might indicate that the WB technique used for these studies could be more sensitive than the IHC method applied. However, this difference could also be attributable to the sampling of different portions of obex used for IHC and WB analyses. The WB molecular mass was similar in all cattle inoculated with CWDwtd and similar to that of cattle-passaged CWDmd (Fig. 3). However, PrPd isoforms from deer infected with CWDwtd had a higher molecular mass (Fig. 3).

This study indicates a high susceptibility of cattle to the CWDwtd agent by the most direct route of inoculation of the CNS, but it remains to be shown if natural routes of exposure would result in infection and disease and whether differences in attack rates and incubation periods between cattle inoculated with CWDmd and CWDwtd agents reflect true differences in susceptibility or are attributable to differences in experimental factors.

Although susceptibility of cattle to intracerebral inoculation of CWDwtd was demonstrated, it should be noted that this is an unnatural route, and it suggests only a potential for cattle to become infected under natural conditions of exposure. It is likely that transmission of CWD to cattle by a more natural route, such as per os, would require a much larger dose of inoculum and might even be difficult to accomplish within the normal life span of the animal. Because the present study resulted in a higher attack rate and shorter incubation periods than similar inoculation with CWDmd, the CWDwtd agent would be the initial choice for experimental oral inoculation of cattle.

Further transmission studies of CWD isolates (e.g., CWD of elk or moose in cattle) are required to explore possible differences in clinicopathologic features that might indicate different disease phenotypes, which may in turn, reflect different CWD agent strains.

Finally, the findings of this study suggest that diagnostic techniques presently used for confirming BSE (IHC and WB) would also detect CWDmd and CWDwtd in cattle should it occur naturally.

http://www.vetpathology.org/cgi/content/full/44/4/487



Vet Pathol 44:487--493 (2007) Susceptibility of Cattle to First-passage Intracerebral Inoculation with Chronic Wasting Disease Agent from White-tailed Deer A. N. HAMIR,J. M. MILLER,R. A. KUNKLE,S. M. HALL,ANDJ. A. RICHT National Animal Disease Center, ARS, USDA, Ames, IA (ANR, JMM, RAK, JAR); and Pathobiology Laboratory, National Veterinary Services Laboratories, Ames, IA (SMR)

http://ddr.nal.usda.gov/dspace/bitstream/10113/3137/1/IND43929871.pdf



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Research Project: Transmission, Differentiation, and Pathobiology of Transmissible Spongiform Encephalopathies Location: Virus and Prion Research Unit

Title: Susceptibility of Cattle to First-Passage Intracerebral Inoculation with Chronic Wasting Disease Agent from Elk

Authors

Greenlee, Justin Nicholson, Eric Kunkle, Robert Hamir, Amirali

Submitted to: American College of Veterinary Pathologists Meeting Publication Type: Abstract Publication Acceptance Date: July 12, 2009 Publication Date: N/A

Technical Abstract: Cattle could be exposed to chronic wasting disease (CWD) from infected farmed or free-ranging cervids. The purpose of this study was to assess the transmissibility of CWD derived from elk to cattle. Intracerebral inoculation of calves (n=14) of approximately 3 months of age was done with 1 ml of a 10% brain homogenate derived from farmed elk with CWD to determine the potential for transmission and define the clinicopathologic features of disease. Non-inoculated calves (n=5) were maintained as controls. Cattle were observed twice daily and necropsies were performed as clinical signs occurred or at the termination of experiment (49 months). Clinical signs of poor appetite, weight loss, circling and bruxism occurred in two cattle (14%) at 16 and 17 months post-inoculation, respectively. The rate of transmission was lower than in cattle inoculated with CWD derived from mule deer (38%) or White-tailed deer (86%). Accumulation of abnormal prion protein (PrPd) in these cattle was confined to the central nervous system and was similar in distribution to cattle inoculated with CWD from mule deer with the most prominent immunoreactivity in midbrain, brainstem, and hippocampus with lesser immunoreactivity in the cervical spinal cord. Additional studies are required to fully assess the potential for cattle to develop CWD through a more natural route of exposure, but a low rate of transmission after intracerebral inoculation suggests that risk of transmission through other routes is low.

http://www.ars.usda.gov/research/publications/publications.htm?seq_no_115=241305



SECOND PASSAGE CWD TO CATTLE ;

doi:10.1016/j.jcpa.2005.07.001

Published by Elsevier Ltd.

Experimental Second Passage of Chronic Wasting Disease (CWDmule deer) Agent to Cattle

A.N. Hamir, R.A. Kunkle, J.M. Miller, J.J. Greenlee and J.A. Richt

Agricultural Research Service, United States Department of Agriculture, National Animal Disease Center, 2300 Dayton Avenue, P.O. Box 70, Ames, IA 50010, USA

Received 4 April 2005; accepted 23 July 2005. Available online 18 January 2006.

Summary To compare clinicopathological findings in first and second passage chronic wasting disease (CWDmule deer) in cattle, six calves were inoculated intracerebrally with brain tissue derived from a first-passage CWD-affected calf in an earlier experiment. Two uninoculated calves served as controls. The inoculated animals began to lose both appetite and weight 10-12 months later, and five subsequently developed clinical signs of central nervous system (CNS) abnormality. By 16.5 months, all cattle had been subjected to euthanasia because of poor prognosis. None of the animals showed microscopical lesions of spongiform encephalopathy (SE) but PrPres was detected in their CNS tissues by immunohistochemistry (IHC) and rapid Western blot (WB) techniques. Thus, intracerebrally inoculated cattle not only amplified CWD PrPres from mule deer but also developed clinical CNS signs in the absence of SE lesions. This situation has also been shown to occur in cattle inoculated with the scrapie agent. The study confirmed that the diagnostic techniques currently used for diagnosis of bovine spongiform encephalopathy (BSE) in the US would detect CWD in cattle, should it occur naturally. Furthermore, it raised the possibility of distinguishing CWD from BSE in cattle, due to the absence of neuropathological lesions and to a distinctive multifocal distribution of PrPres, as demonstrated by IHC which, in this study, appeared to be more sensitive than the WB technique.

Keywords: cattle; chronic wasting disease (CWD); deer; transmissible spongiform encephalopathy (TSE)

SNIP...

Discussion CWD, like all other TSEs, is characterized by a long incubation period, which in deer is seldom less than 18 months (Williams and Young, 1992). In an experimental study of cattle inoculated intracerebrally with CWD from mule deer (first passage), amplification of PrPres was demonstrated in only five of 13 (38%) cattle, after incubation periods that ranged from 23 to 63 months (Hamir et al., 2001a, 2005a). In contrast, all inoculated cattle in the present study were positive for PrPres within 16.5 months. This increased attack rate with shorter incubation periods probably indicates adaptation of the CWDmule deer agent to a new host. It could also be argued that the inoculum used for the primary passage (Hamir et al., 2001a, 2005a) had a lower infectivity titre than that used for the second passage. However, the former successfully transmitted CWD to each of five white tailed deer within two years of intracerebral inoculation (Kunkle et al., Unpublished).

In cervids, clinical CWD is characterized by emaciation, changes in behaviour, and excessive salivation (Williams and Young, 1992). Although the latter was not observed in the CWD inoculated cattle, all animals showed anorexia and considerable weight loss. Five cattle also showed intermittent neurological signs. Although none of these animals showed histopathological changes in the brain, all were shown to be positive for PrPres by the IHC and WB methods. The presence of isolated vacuoles in the red nucleus is regarded as an incidental finding in cattle (McGill and Wells, 1993). The uniform susceptibility, relatively short incubation, and absence of microscopical lesions in cattle given CWD brain material passaged once through cattle resembled findings in cattle inoculated intracerebrally with the scrapie agent (Cutlip et al., 1997). In that experiment, 100% of cattle died 14-18 months after inoculation with material from the first cattle-passage of a US strain of the scrapie agent; none showed microscopical lesions and all were positive for PrPres. In the present experiment, the possibility that the PrPres seen in tissue sections represented residual CWD material from the inoculum was ruled out because of the multifocal distribution of

PrPres throughout the brain (excluding cerebellar folia) and cervical spinal cord of most of the affected animals. Had the PrPres represented residual inoculum, it would probably have been confined to the sites of deposition in the midbrain or cerebrum. Moreover, in studies on sheep scrapie, Hamir et al. (2002) showed that intracerebrally inoculated brain material containing PrPres was present for only a few days in sufficient quantity to be detectable immunohistochemically. The present work confirms previous observations that PrPres IHC labelling in cattle inoculated with the mule deer CWD agent is multifocal and glial cell-associated. This unusual pattern was first reported in descriptions of the primary CWD transmission to cattle (Hamir et al., 2001a, 2005a), and the study described here showed that it was maintained through the second passage in cattle. Further studies now in progress will determine whether this feature also characterizes CWD transmission to cattle fromother cervid species other than mule deer, namely, white tailed deer and elk. In this and an earlier study of CWD in cattle (Hamir et al., 2001a), IHC labelling differed from that seen in cattle with BSE or experimental transmissible mink encephalopathy (TME), both of which are associated with widespread diffuse labelling of grey matter neuropil, with labelled particles that are not obviously cell-associated except occasionally at neuronal cell membranes (Wells and Willsmith, 1995; Hamir et al., 2005a). The IHC pattern in bovine CWD also contrasts markedly with that seen in scrapie-inoculated cattle, in which intracytoplasmic labelling of neurons is a prominent feature (Cutlip et al., 1994, 1997). When brainstems of CWD-infected cattle were analysed by WB for the presence of PrPres, only three of six samples were found to be positive (Table 1). In contrast, all samples from the midbrain area were positive by this technique (Table 1; Fig. 5). It was noteworthy, however, that both brainstem and midbrain sections of all animals infected with CWD gave positive IHC results (Table 1) and a positive WB was associated with strong IHC labelling. This may indicate that the IHC procedure is more sensitive than the WB method for cattle-passaged CWD. However, given the multifocal nature of PrPres distribution in the CNS of CWD-infected cattle, this result is not surprising. WB analysis requires a small sample of brain tissue (e.g. 0.2 g, as in the present study) to produce a 10% homogenate; approximately 10 ml (1 mg brain tissue equivalent) of this homogenate are loaded on to an SDS-PAGE gel for further

analysis. Bearing in mind the multifocal pattern of PrPres distribution, the brain tissue used for the preparation of WB homogenate, unlike the large amount examined in the IHC procedure, might well contain few if any foci of PrPres deposition, whereas the larger piece of tissue section used for IHC may contain detectable PrPres. In this respect, therefore, the IHC method would seem preferable to the WB procedure and to other procedures (e.g. ELISA-based tests) in which only small amounts of tissue are used for analysis. In comparison with experimental TME in cattle (Hamir et al., 2005b), the experimental bovine CWD in this study was associated with less extensive IHC labelling in non-CNS (i.e. other than brain and spinal cord) neural tissues. Whereas the retina was positive in all cattle inoculated with TME, none of the CWD-infected cattle in this experiment had any retinal labelling. Similarly, in the present study there was no labelling in the pituitary gland, a tissue sometimes positive in TME-infected cattle. Because the incubation time for second passage CWD transmission (mean of 468 days) was only slightly longer than for TME (mean of 430 days), it seems likely that these different tissue affinities reflect a biological difference between these two TSE agents. PrPres IHC labelling was not observed in striated muscles (heart, tongue, masseter, diaphragm) of the experimental animals. This observation accorded with our previous findings (Hamir et al., 2004a) in which striated muscle tissues from 20 animals (cattle, sheep, elk and raccoons) were examined for PrPres. In these animals, all of which had developed a TSE after experimental inoculation, PrPres was found by IHC examination in the brains, but not in muscle tissues. However, recent investigations with an enriched WB technique (Mulcahy et al., 2004) have enabled us to detect PrPres in the tongues of some sheep and elk experimentally inoculated with scrapie and CWD, respectively. This technique failed, however, to detect PrPres in cattle inoculated with CWD or TME (Bessen et al., unpublished). This study is still in progress, and the tongues of TSE-infected animals are currently being tested after careful removal from the carcasses to ensure non-contamination with infected brain material. The present study and a previous experiment (Hamir et al., 2005a) have established the biological characteristics of the CWDmule deer agent in cattle. However, isolates of CWD from other cervids (e.g. CWDwhite-tailed and CWDelk) may differ. Transmission experiments with different CWD isolates are therefore needed to examine the possibility of variation in the CWD agent in wild cervids. Such experiments have recently been initiated at the National Animal Disease Center (NADC). Acknowledgments We thank Dr Katherine I. O'Rourke for providing the antibody for the IHC procedure. Martha Church, Kevin Hassall, Dennis Orcutt, Jean Donald, Sharla Van Roekel, and animal handlers at the NADC provided expert technical assistance. This study was carried out under the guidelines of the institutional Animal Care and Use committee at NADC. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the United States Department of Agriculture.

ABSTRACT

http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WHW-4J2N7WS-3&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_searchStrId=1049833173&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=0980bd13d461337321f0656ec3b56863



FULL TEXT ;

http://www.cwd.cc/Experimental%20Second%20Passage%20of%20Chronic%20Wasting%20Disease%20Agent%20to%20Cattle.pdf



ALSO SEE ;

Susceptibility of Domestic Cattle to Chronic Wasting Disease by Oral Inoculation and Natural Exposure: Final Phase of a 10-year Study

Elizabeth S. Williams1, Donal O'Toole1, Matthew M. Hille1, Donald L. Montgomery1, Jean E. Jewell1*, Terry J. Kreeger2, and Michael W. Miller3 1Department of Veterinary Sciences, University of Wyoming, Laramie, WY 82070 2Wyoming Game and Fish Department, Wheatland, WY 82201 3Colorado Division of Wildlife, Fort Collins, Colorado 80526 *Corresponding author e-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000164/!x-usc:mailto:jjewell@uwyo.edu The risk of domestic cattle developing a transmissible spongiform encephalopathy (TSE) after oral inoculation followed by a long incubation period, or by long-term natural exposure to cervids infected with chronic wasting disease (CWD) was studied. Ten cattle were given large oral doses of pooled brain material from CWD-infected mule deer in late August 1997 and housed in isolation at the Wyoming State Veterinary Laboratory until September 2007. Two additional groups of cattle were penned outdoors with CWD-infected deer and elk or in CWD-contaminated premises at Colorado Division of Wildlife (n=11) and Wyoming Game and Fish Department (n=10) research facilities during the same ten-year period. These conditions simulated exposure routes that cattle in North America might encounter if they are raised or grazed in areas where free-ranging or captive deer and elk are infected with CWD. Beginning in July 2007 all exposed and three untreated control cattle were killed, and select tissues were collected at necropsy. Samples from each animal were analyzed for the diagnostic hallmarks of TSEs by immunohistochemistry and Western blot. DNA sequences were determined for the cellular prion protein gene in each animal. No proteinase-K resistant prion protein or anti-PrP immunoreactive IHC signals were detected in any tissues of exposed or control animals. None of these results, taken individually or together, support a diagnosis of TSE in cattle inoculated orally with a high dose of infectious CWD material or continually exposed by cohabitation with infected deer or elk, or transmission from contaminated premises despite an incubation period of up to 10 or 11 years.

http://www.cwd-info.org/pdf/3rd_CWD_Symposium_utah.pdf




Detection of protease-resistant cervid prion protein in water from a CWD-endemic area

T.A. Nichols,1,2 Bruce Pulford,1 A. Christy Wyckoff,1,2 Crystal Meyerett,1 Brady Michel,1 Kevin Gertig,3 Edward A. Hoover,1 Jean E. Jewell,4 Glenn C. Telling5 and Mark D. Zabel1,*

1Department of Microbiology, Immunology and Pathology; College of Veterinary Medicine and Biomedical Sciences; Colorado State University; Fort Collins, CO USA; 2National Wildlife Research Center; Wildlife Services; United States Department of Agriculture; Fort Collins, CO USA; 3Fort Collins Utilities; Fort Collins; CO USA; 4Department of Veterinary Sciences; Wyoming State Veterinary Laboratory; University of Wyoming; Laramie, WY USA; 5Department of Microbiology, Immunology, Molecular Genetics and Neurology; Sanders Brown Center on Aging; University of Kentucky; Lexington, KY USA

Key words: prions, chronic wasting disease, water, environment, serial protein misfolding cyclic amplification Abbreviations: CWD, chronic wasting disease; sPMCA, serial protein misfolding cyclic amplification; PrPC, cellular prion protein; PrPSc, disease-related, misfolded murine PrP; PrPCWD, disease-related, misfolded cervid PrP; PrPRES, protease-resistant PrP; FCWTF, Fort Collins water treatment facility

Chronic wasting disease (CWD) is the only known transmissible spongiform encephalopathy affecting free-ranging wildlife. Although the exact mode of natural transmission remains unknown, substantial evidence suggests that prions can persist in the environment, implicating components thereof as potential prion reservoirs and transmission vehicles.1-4 CWD-positive animals may contribute to environmental prion load via decomposing carcasses and biological materials including saliva, blood, urine and feces.5-7 Sensitivity limitations of conventional assays hamper evaluation of environmental prion loads in soil and water. Here we show the ability of serial protein misfolding cyclic amplification (sPMCA) to amplify a 1.3 x 10-7 dilution of CWD-infected brain homogenate spiked into water samples, equivalent to approximately 5 x 107 protease resistant cervid prion protein (PrPCWD) monomers. We also detected PrPCWD in one of two environmental water samples from a CWD endemic area collected at a time of increased water runoff from melting winter snow pack, as well as in water samples obtained concurrently from the flocculation stage of water processing by the municipal water treatment facility. Bioassays indicated that the PrPCWD detected was below infectious levels. These data demonstrate detection of very low levels of PrPCWD in the environment by sPMCA and suggest persistence and accumulation of prions in the environment that may promote CWD transmission.

snip...

The data presented here demonstrate that sPMCA can detect low levels of PrPCWD in the environment, corroborate previous biological and experimental data suggesting long term persistence of prions in the environment2,3 and imply that PrPCWD accumulation over time may contribute to transmission of CWD in areas where it has been endemic for decades. This work demonstrates the utility of sPMCA to evaluate other environmental water sources for PrPCWD, including smaller bodies of water such as vernal pools and wallows, where large numbers of cervids congregate and into which prions from infected animals may be shed and concentrated to infectious levels.

http://www.landesbioscience.com/journals/prion/NicholsPRION3-3.pdf



Monday, August 24, 2009 Third International CWD Symposium July 22-24, 2009 - Park City, Utah ABSTRACTS

http://chronic-wasting-disease.blogspot.com/2009/08/third-international-cwd-symposium-july.html




Thursday, October 15, 2009

Transmissibility studies of vacuolar changes in the rostral colliculus of pigs


http://madporcinedisease.blogspot.com/2009/10/transmissibility-studies-of-vacuolar.html




http://madporcinedisease.blogspot.com/




FELINE SPONGIFORM ENCEPHALOPATHY FSE


http://felinespongiformencephalopathyfse.blogspot.com/




2005 DEFRA Department for Environment, Food & Rural Affairs

Area 307, London, SW1P 4PQ Telephone: 0207 904 6000 Direct line: 0207 904 6287 E-mail: h.mcdonagh.defra.gsi.gov.uk

GTN: FAX:

Mr T S Singeltary P.O. Box 42 Bacliff Texas USA 77518

21 November 2001

Dear Mr Singeltary

TSE IN HOUNDS

Thank you for e-mail regarding the hounds survey. I am sorry for the long delay in responding.

As you note, the hound survey remains unpublished. However the Spongiform Encephalopathy Advisory Committee (SEAC), the UK Government’s independent Advisory Committee on all aspects related to BSE-like disease, gave the hound study detailed consideration at their meeting in January 1994. As a summary of this meeting published in the BSE inquiry noted, the Committee were clearly concerned about the work that had been carried out, concluding that there had clearly been problems with it, particularly the control on the histology, and that it was more or less inconclusive. However was agreed that there should be a re-evaluation of the pathological material in the study.

Later, at their meeting in June 95, The Committee re-evaluated the hound study to see if any useful results could be gained from it. The Chairman concluded that there were varying opinions within the Committee on further work. It did not suggest any further transmission studies and thought that the lack of clinical data was a major weakness.

Overall, it is clear that SEAC had major concerns about the survey as conducted. As a result it is likely that the authors felt that it would not stand up to r~eer review and hence it was never published. As noted above, and in the detailed minutes of the SEAC meeting in June 95, SEAC considered whether additional work should be performed to examine dogs for evidence of TSE infection. Although the Committee had mixed views about the merits of conducting further work, the Chairman noted that when the Southwood Committee made their recommendation to complete an assessment of possible spongiform disease in dogs, no TSEs had been identified in other species and hence dogs were perceived as a high risk population and worthy of study. However subsequent to the original recommendation, made in 1990, a number of other species had been identified with TSE ( e.g. cats) so a study in hounds was less

critical. For more details see-



http://www.bseinquiry.gov.uk/files/yb/1995/06/21005001.pdf




As this study remains unpublished, my understanding is that the ownership of the data essentially remains with the original researchers. Thus unfortunately, I am unable to help with your request to supply information on the hound survey directly. My only suggestion is that you contact one of the researchers originally involved in the project, such as Gerald Wells. He can be contacted at the following address.

Dr Gerald Wells, Veterinary Laboratories Agency, New Haw, Addlestone, Surrey, KT 15 3NB, UK

You may also wish to be aware that since November 1994 all suspected cases of spongiform encephalopathy in animals and poultry were made notifiable. Hence since that date there has been a requirement for vets to report any suspect SE in dogs for further investigation. To date there has never been positive identification of a TSE in a dog.

I hope this is helpful

Yours sincerely 4

HUGH MCDONAGH BSE CORRESPONDENCE SECTION

IN CONFIDENCE

CONCEPT NOT FOR FURTHER STUDY OF MATERIAL OBTAINED IN A SURVEY OF HOUNDS FOR EVIDENCE OF A SCRAPIE-LIKE SPONGIFORM ENCEPHALOPATHY (SE)

snip…

b) Fibrillar material closely similar to SAF, found in BSE/Scrapie, was observed in 19 (4.3%) cases, all of which were hounds > 7 years of age. 14/19 of these suspected SAF results correlated with cases in the unresolveable histopathological category.

snip…

The following proposals address the hypothesis that the hound survey observations represent a PrP related or scrapie-like disease of dogs in which the pathological response, and possible the spread of infectivity, is neuroanatomically localized. By inference this could also mean that the disorder is clinically silent and non-progressive.



http://www.bseinquiry.gov.uk/files/yb/1995/02/09001001.pdf




http://www.mad-cow.org/00/aug00_late_news.html#ggg




37.Putative TSE in hounds - work started 1990 -(see para 41)

Robert Higgins, a Veterinary Investigation Officer at Thirsk, had been working on a hound survey in 1990. Gerald Wells and I myself received histological sections from this survey along with the accompanying letter (YB90/11.28/1.1) dated November 1990. This letter details spongiform changes found in brains from hunt hounds failing to keep up with the rest of the pack, along with the results of SAF extractions from fresh brain material from these same animals. SAFs were not found in brains unless spongiform changes were also present. The spongiform changes were not pathognomonic (ie. conclusive proof) for prion disease, as they were atypical, being largely present in white matter rather than grey matter in the brain and spinal cord. However, Tony Scott, then head of electron microscopy work on TSEs, had no doubt that these SAFs were genuine and that these hounds therefore must have had a scrapie-like disease. I reviewed all the sections myself (original notes appended) and although the pathology was not typical, I could not exclude the possibility that this was a scrapie-like disorder, as white matter vacuolation is seen in TSEs and Wallerian degeneration was also present in the white matter of the hounds, another feature of scrapie.

38.I reviewed the literature on hound neuropathology, and discovered that micrographs and descriptive neuropathology from papers on 'hound ataxia' mirrored those in material from Robert Higgins' hound survey. Dr Tony Palmer (Cambridge) had done much of this work, and I obtained original sections from hound ataxia cases from him. This enabled me provisionally to conclude that Robert Higgins had in all probability detected hound ataxia, but also that hound ataxia itself was possibly a TSE. Gerald Wells confirmed in 'blind' examination of single restricted microscopic fields that there was no distinction between the white matter vacuolation present in BSE and scrapie cases, and that occurring in hound ataxia and the hound survey cases.

39.Hound ataxia had reportedly been occurring since the 1930's, and a known risk factor for its development was the feeding to hounds of downer cows, and particularly bovine offal. Circumstantial evidence suggests that bovine offal may also be causal in FSE, and TME in mink. Despite the inconclusive nature of the neuropathology, it was clearly evident that this putative canine spongiform encephalopathy merited further investigation.

40.The inconclusive results in hounds were never confirmed, nor was the link with hound ataxia pursued. I telephoned Robert Higgins six years after he first sent the slides to CVL. I was informed that despite his submitting a yearly report to the CVO including the suggestion that the hound work be continued, no further work had been done since 1991. This was surprising, to say the very least.

41.The hound work could have provided valuable evidence that a scrapie-like agent may have been present in cattle offal long before the BSE epidemic was recognised. The MAFF hound survey remains unpublished.

Histopathological support to various other published MAFF experiments

42.These included neuropathological examination of material from experiments studying the attempted transmission of BSE to chickens and pigs (CVL 1991) and to mice (RVC 1994).



http://www.bseinquiry.gov.uk/witness/htm/stat067.htm




nothing to offer scientifically;



http://www.bseinquiry.gov.uk/files/yb/1991/10/17001001.pdf




maddogs and Englishman



http://www.bseinquiry.gov.uk/files/yb/1990/11/28001001.pdf




kind regards, terry


###########bse-l ############


Subject: Re: DEER SPONGIFORM ENCEPHALOPATHY SURVEY & HOUND STUDY
Date: Fri, 18 Oct 2002 23:12:22 +0100
From: Steve Dealler
Reply-To: Bovine Spongiform Encephalopathy Organization: Netscape Online member T
o: BSE-L@ References: <mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000164/!x-usc:mailto:3DAF5023.4080804@wt.net>

Dear Terry,

An excellent piece of review as this literature is desparately difficult to get back from Government sites.

What happened with the deer was that an association between deer meat eating and sporadic CJD was found in about 1993. The evidence was not great but did not disappear after several years of asking CJD cases what they had eaten. I think that the work into deer disease largely stopped because it was not helpful to the UK industry...and no specific cases were reported. Well, if you dont look adequately like they are in USA currenly then you wont find any!

Steve Dealler ===============

Incubation periods for BSE are proportional to the life expectancy of the animal affected. The disease's incubation period is 18% of a cow's life expectancy and would be expected to about double when crossing to another species [---] that is, to 36% of 70 years in humans.

Steve Dealler, consultant in medical microbiology. Burnley General Hospital, Burnley BB10 2PQ mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000164/!x-usc:mailto:deal@airtime.co.uk

TSS

########### http://mailhost.rz.uni-karlsruhe.de/warc/bse-l.html ############

see ;


http://felinespongiformencephalopathyfse.blogspot.com/




P.9.1

Surveillance for Prion diseases in the United States

Robert Holman, Ermias Belay, Krista Christensen, Ryan Maddox, Arialdi Minino, Arianne Folkema, Dana Haberling, Teresa Hammett, Kenneth Kochanek, Lawrence Schonberger CDC, USA

Background: Prion diseases are a family of rare progressive neurodegenerative disorders that affect humans and animals. The most common form of human prion disease, Creutzfeldt- Jakob disease (CJD), is believed to occur in many countries of the world. Variant CJD (vCJD), a recently emerged human prion disease, has been causally associated with bovine spongiform encephalopathy.

Objectives: To describe the occurrence of CJD and vCJD in the United States.

Methods: Analysis of prion disease deaths on death certificates of US residents, 1979-2006, and vCJD deaths identified through other surveillance mechanisms, 1996-2008. Since CJD is invariably fatal and illness duration is usually less than one year, the CJD incidence is estimated as the death rate.

Results: During 1979 through 2006, 6911 deaths with CJD listed as a cause were reported in the United States, an annual average of approximately 247 deaths (range 172-304 deaths). The average annual age-adjusted incidence for CJD was 0.97 per million persons (95% CI=0.95-0.99). Most (61.8%) of these cases occurred among persons
65 years of age for an average annual incidence in this age-group of 4.8 per million persons. Most cases were among whites (94.6%); the age-adjusted incidence for whites was >2.5 times higher than that for blacks (1.04 and 0.40, respectively). Three patients who died since 2004 were reported with vCJD; epidemiological evidence indicated that their infection was acquired outside of the United States.

Discussion: National surveillance continues to show an annual CJD incidence rate of about 1 case per million persons and marked differences in CJD rates by age and race in the United States. Ongoing surveillance remains important for monitoring the stability of the CJD incidence rates, and detecting occurrences of vCJD and possibly other novel prion diseases in the United States.

P.9.18

Neuropathology-confirmed CJD decedents less than 55 years of age, United States, 1994-2006

Ryan Maddox1, Robert Holman1, Pierluigi Gambetti2, Janis Blevins2, Sally Berri2, Arialdi Minino1, Krista Christensen1, James Sejvar1, Lawrence Schonberger1, Ermias Belay1 1Centers for Disease Control and Prevention, USA; 2National Prion Disease Pathology Surveillance Center, USA

Background: Approximately 10% of US Creutzfeldt-Jakob disease (CJD) decedents are reported to be <55 href="http://www.prion2009.com/sites/default/files/Prion2009_Book_of_Abstracts.pdf">http://www.prion2009.com/sites/default/files/Prion2009_Book_of_Abstracts.pdf




IN my opinion, all human TSE, OF ALL AGE GROUPS, must be reported in every State and Nation. THEY MUST BE REPORABLE if not for the following reason alone. IATROGENIC CREUTZFELDT JAKOB DISEASE knows no age group. The 'pass it forward' and or 'friendly fired' mode of transmission with TSE should be enough alone to manadory all human TSE reportable in every State, of any age. ...TSS

Tuesday, August 11, 2009 Characteristics of Established and Proposed Sporadic Creutzfeldt-Jakob Disease Variants

Brian S. Appleby, MD; Kristin K. Appleby, MD; Barbara J. Crain, MD, PhD; Chiadi U. Onyike, MD, MHS; Mitchell T. Wallin, MD, MPH; Peter V. Rabins, MD, MPH

Background: The classic Creutzfeldt-Jakob disease (CJD), Heidenhain, and Oppenheimer-Brownell variants are sporadic CJD (sCJD) phenotypes frequently described in the literature, but many cases present with neuropsychiatric symptoms, suggesting that there may be additional sCJD phenotypes.

Objective: To characterize clinical, diagnostic, and molecular features of 5 sCJD variants.

Design: Retrospective analysis.

Setting: The Johns Hopkins and Veterans Administration health care systems.

Participants: Eighty-eight patients with definite or probable sCJD.

Main Outcome Measures: Differences in age at onset, illness progression, diagnostic test results, and molecular subtype.

Results: The age at onset differed among sCJD variants (P=.03); the affective variant had the youngest mean age at onset (59.7 years). Survival time (P
.001) and the time to clinical presentation (P=.003) differed among groups. Patients with the classic CJD phenotype had the shortest median survival time from symptom onset (66 days) and those who met criteria for the affective sCJD variant had the longest (421 days) and presented to clinicians significantly later (median time from onset to presentation, 92 days; P=.004). Cerebrospinal fluid analyses were positive for 14-3-3 protein in all of the affective variants, regardless of illness duration. Periodic sharp-wave complexes were not detected on any of the electroencephalography tracings in the Oppenheimer-Brownell group; basal ganglia hyperintensity was not detected on brain magnetic resonance imaging in this group either. All of the Heidenhain variants were of the methionine/ methionine type 1 molecular subtype.

Conclusions: The classic CJD phenotype and the Heidenhain, Oppenheimer-Brownell, cognitive, and affective sCJD variants differ by age at disease onset, survival time, and diagnostic test results. Characteristics of these 5 phenotypes are provided to facilitate further clinicopathologic investigation that may lead to more reliable and timely diagnoses of sCJD.

Arch Neurol. 2009;66(2):208-215


http://archneur.ama-assn.org/



http://creutzfeldt-jakob-disease.blogspot.com/2009/08/characteristics-of-established-and.html




Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States 2003 revisited 2009

August 10, 2009

Greetings,

I would like to submit a review of past CJD surveillance in the USA, and the urgent need to make all human TSE in the USA a reportable disease, in every state, of every age group, and to make this mandatory immediately without further delay. The ramifications of not doing so will only allow this agent to spread further in the medical, dental, surgical arena’s. North America seems to have the most species with documented Transmissible Spongiform Encephalopathy's, most all of which have been rendered and fed back to food producing animals and to humans for years. If you look at the statistics, sporadic CJD seems to be rising in the USA, and has been, with atypical cases of the sCJD. I find deeply disturbing in the year of 2009, that Human Transmissible Spongiform Encephalopathy of any strain and or phenotype, of all age groups, and I stress all age groups, because human TSE's do not know age, and they do not know borders. someone 56 years old, that has a human TSE, that has surgery, can pass this TSE agent on i.e. friendly fire, and or passing it forward, and there have been documented nvCJD in a 74 year old. Remembering also that only sporadic CJD has been documented to transmit via iatrogenic routes, until recently with the 4 cases of blood related transmission, of which the origin is thought to be nvCJD donors. However most Iatrogenic CJD cases are nothing more than sporadic CJD, until the source is proven, then it becomes Iatrogenic. An oxymoron of sorts, because all sporadic CJD is, are multiple forms, or strains, or phenotypes of Creutzfeldt Jakob Disease, that the route and source and species have not been confirmed and or documented. When will the myth of the UKBSEnvCJD only theory be put to bed for good. This theory in my opinion, and the following there from, as the GOLD STANDARD, has done nothing more than help spread this agent around the globe. Politics and money have caused the terrible consequences to date, and the fact that TSEs are a slow incubating death, but a death that is 100% certain for those that are exposed and live long enough to go clinical. once clinical, there is no recourse, to date. But, while sub-clinical, how many can one exposed human infect? Can humans exposed to CWD and scrapie strains pass it forward as some form of sporadic CJD in the surgical and medical arenas? why must we wait decades and decades to prove this point, only to expose millions needlessly, only for the sake of the industries involved? would it not have been prudent from the beginning to just include all TSE's, and rule them out from there with transmission studies and change policies there from, as opposed to doing just the opposite? The science of TSE's have been nothing more than a political circus since the beginning, and for anyone to still believe in this one strain, one group of bovines, in one geographical location, with only one age group of human TSE i.e. nvCJD myth, for anyone to believe this today only enhances to spreading of these human and animal TSE's. This is exactly why we have been in this quagmire.

The ones that believe that there is a spontaneous CJD in 85%+ of all cases of human TSE, and the ones that do not believe that cattle can have this same phenomenon, are two of the same, the industry, and so goes the political science aspect of this tobacco and or asbestos scenario i.e. follow the money. I could go into all angles of this man made nightmare, the real facts and science, for instance, the continuing rendering technology and slow cooking with low temps that brewed this stew up, and the fact that THE USA HAD THIS TECHNOLOGY FIRST AND SHIPPED IT TO THE U.K. SOME 5 YEARS BEFORE THE U.S. STARTED USING THE SAME TECHNOLOGY, to save on fuel cost. This is what supposedly amplified the TSE agent via sheep scrapie, and spread via feed in the U.K. bovine, and other countries exporting the tainted product. BUT most everyone ignores this fact, and the fact that the U.S. has been recycling more TSE, from more species with TSEs, than any other country documented, but yet, it's all spontaneous, and the rise in sporadic CJD in the U.S. is a happenstance of bad luck ??? I respectfully disagree. To top that all off, the infamous BSE-FIREWALL that the USDA always brags about was nothing more than ink on paper, and I can prove this. YOU can ignore it, but this is FACT (see source, as late as 2007, in one recall alone, some 10,000,000 MILLION POUNDS OF BANNED MAD COW FEED WENT OUT INTO COMMERCE TO BE FED OUT, and most was never recovered. This was banned blood laced, meat and bone meal. 2006 was a banner year for banned mad cow protein going into commerce in the U.S. (see source of FDA feed ban warning letter below). I stress that the August 4, 1997 USA mad cow feed ban and this infamous BSE firewall, was nothing more than ink on paper, it was never enforceable.

I propose that the current diagnostic criteria for human TSEs only enhances and helps the spreading of human TSE from the continued belief of the UKBSEnvCJD only theory in 2009. With all the science to date refuting it, to continue to validate this old myth, will only spread this TSE agent through a multitude of potential routes and sources i.e. consumption, medical i.e., surgical, blood, dental, endoscopy, optical, nutritional supplements, cosmetics etc. I propose as with Aguzzi, Asante, Collinge, Caughey, Deslys, Dormont, Gibbs, Gajdusek, Ironside, Manuelidis, Marsh, et al and many more, that the world of TSE Transmissible Spongiform Encephalopathy is far from an exact science, but there is enough proven science to date that this myth should be put to rest once and for all, and that we move forward with a new classification for human and animal TSE that would properly identify the infected species, the source species, and then the route. This would further have to be broken down to strain of species and then the route of transmission would further have to be broken down. Accumulation and Transmission are key to the threshold from sub- clinical to clinical disease, and key to all this, is to stop the amplification and transmission of this agent, the spreading of, no matter what strain. In my opinion, to continue with this myth that the U.K. strain of BSE one strain TSE in cows, and the nv/v CJD one strain TSE humans, and the one geographical location source i.e. U.K., and that all the rest of human TSE are just one single strain i.e. sporadic CJD, a happenstance of bad luck that just happens due to a twisted protein that just twisted the wrong way, IN 85%+ OF ALL HUMAN TSEs, when to date there are 6 different phenotypes of sCJD, and growing per Gambetti et al, and that no other animal TSE transmits to humans ??? With all due respect to all Scientist that believe this, I beg to differ. To continue with this masquerade will only continue to spread, expose, and kill, who knows how many more in the years and decades to come. ONE was enough for me, My Mom, hvCJD i.e. Heidenhain Variant CJD, DOD 12/14/97 confirmed, which is nothing more than another mans name added to CJD, like CJD itself, Jakob and Creutzfeldt, or Gerstmann-Straussler-Scheinker syndrome, just another CJD or human TSE, named after another human. WE are only kidding ourselves with the current diagnostic criteria for human and animal TSE, especially differentiating between the nvCJD vs the sporadic CJD strains and then the GSS strains and also the FFI fatal familial insomnia strains or the ones that mimics one or the other of those TSE? 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. ...

please see history, and the ever evolving TSE science to date ;


Saturday, June 13, 2009

Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States 2003 revisited 2009


http://cjdusa.blogspot.com/2009/06/monitoring-occurrence-of-emerging-forms.html




JOURNAL OF NEUROLOGY

MARCH 26, 2003

Send Post-Publication Peer Review to journal:

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

disease in the United States

Email Terry S. Singeltary:

mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000748/!x-usc:mailto:flounder@wt.net

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?


http://www.neurology.org/cgi/eletters/60/2/176#535




LANCET INFECTIOUS DISEASE JOURNAL

Volume 3, Number 8 01 August 2003

Newsdesk

Tracking spongiform encephalopathies in North America

Xavier Bosch

My name is Terry S Singeltary Sr, and I live in Bacliff, Texas. I lost my mom to hvCJD (Heidenhain variant CJD) and have been searching for answers ever since. What I have found is that we have not been told the truth. CWD in deer and elk is a small portion of a much bigger problem.

49-year-old Singeltary is one of a number of people who have remained largely unsatisfied after being told that a close relative died from a rapidly progressive dementia compatible with spontaneous Creutzfeldt-Jakob disease (CJD). So he decided to gather hundreds of documents on transmissible spongiform encephalopathies (TSE) and realised that if Britons could get variant CJD from bovine spongiform encephalopathy (BSE), Americans might get a similar disorder from chronic wasting disease (CWD)the relative of mad cow disease seen among deer and elk in the USA. Although his feverish search did not lead him to the smoking gun linking CWD to a similar disease in North American people, it did uncover a largely disappointing situation.

Singeltary was greatly demoralised at the few attempts to monitor the occurrence of CJD and CWD in the USA. Only a few states have made CJD reportable. Human and animal TSEs should be reportable nationwide and internationally, he complained in a letter to the Journal of the American Medical Association (JAMA 2003; 285: 733). I hope that the CDC does not continue to expect us to still believe that the 85% plus of all CJD cases which are sporadic are all spontaneous, without route or source.

Until recently, CWD was thought to be confined to the wild in a small region in Colorado. But since early 2002, it has been reported in other areas, including Wisconsin, South Dakota, and the Canadian province of Saskatchewan. Indeed, the occurrence of CWD in states that were not endemic previously increased concern about a widespread outbreak and possible transmission to people and cattle.

To date, experimental studies have proven that the CWD agent can be transmitted to cattle by intracerebral inoculation and that it can cross the mucous membranes of the digestive tract to initiate infection in lymphoid tissue before invasion of the central nervous system. Yet the plausibility of CWD spreading to people has remained elusive.

Part of the problem seems to stem from the US surveillance system. CJD is only reported in those areas known to be endemic foci of CWD. Moreover, US authorities have been criticised for not having performed enough prionic tests in farm deer and elk.

Although in November last year the US Food and Drug Administration issued a directive to state public-health and agriculture officials prohibiting material from CWD-positive animals from being used as an ingredient in feed for any animal species, epidemiological control and research in the USA has been quite different from the situation in the

UK and Europe regarding BSE.

Getting data on TSEs in the USA from the government is like pulling teeth, Singeltary argues. You get it when they want you to have it and only what they want you to have.Norman Foster, director of the Cognitive Disorders Clinic at the University of Michigan (Ann Arbor, MI, USA), says that current surveillance of prion disease in people in the USA is inadequate to detect whether CWD is occurring in human beings; adding that, the cases that we know about are reassuring, because they do not suggest the appearance of a new variant of CJD in the USA or atypical features in patients that might be exposed to CWD. However, until we establish a system that identifies and analyses a high proportion of suspected prion disease cases we will not know for sure. The USA should develop a system modelled on that established in the UK, he points out.

Ali Samii, a neurologist at Seattle VA Medical Center who recently reported the cases of three hunterstwo of whom were friendswho died from pathologically confirmed CJD, says that at present there are insufficient data to claim transmission of CWD into humans; adding that [only] by asking [the questions of venison consumption and deer/elk hunting] in every case can we collect suspect cases and look into the plausibility of transmission further. Samii argues that by making both doctors and hunters more aware of the possibility of prions spreading through eating venison, doctors treating hunters with dementia can consider a possible prion disease, and doctors treating CJD patients will know to ask whether they ate venison. CDC spokesman Ermias Belay says that the CDC will not be investigating the [Samii] cases because there is no evidence that the men ate CWD-infected meat. He notes that although the likelihood of CWD jumping the species barrier to infect humans cannot be ruled out 100% and that [we] cannot be 100% sure that CWD does not exist in humans & the data seeking evidence of CWD transmission to humans have been very limited.


http://www.thelancet.com/journals/laninf/article/PIIS1473309903007151/%20fulltext




he complained in a letter to the Journal of the American Medical Association (JAMA 2003; 285: 733). 

I hope that the CDC does not continue to expect us to still believe that the 85% plus of all CJD cases which are sporadic are all spontaneous, without route or source.<<< href="mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000164/!x-usc:http://jama.ama-assn.org/cgi/content/extract/285/6/733?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=singeltary&searchid=1&FIRSTINDEX=0&resourcetype=HWCIT">http://jama.ama-assn.org/cgi/content/extract/285/6/733?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=singeltary&searchid=1&FIRSTINDEX=0&resourcetype=HWCIT



http://jama.ama-assn.org/cgi/content/full/285/6/733?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=singeltary&searchid=1&FIRSTINDEX=0&resourcetype=HWCIT




2 January 2000

British Medical Journal

U.S. Scientist should be concerned with a CJD epidemic in the U.S., as well


http://www.bmj.com/cgi/eletters/320/7226/8/b#6117




15 November 1999

British Medical Journal

vCJD in the USA * BSE in U.S.


http://www.bmj.com/cgi/eletters/319/7220/1312/b#5406




THE PATHOLOGICAL PROTEIN

BY Philip Yam

Yam Philip Yam News Editor Scientific American http://www.sciam.com/


http://www.thepathologicalprotein.com/




SEE REVISITING SPORADIC CJD BY PHILIP YAM THE PATHOLOGICAL PROTEIN

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


http://books.google.com/books?id=ePbrQNFrHtoC&pg=PA224&lpg=PA224&dq=pathological+protein+philip+yam+singeltary&source=bl&ots=um-LytTT2E&sig=hQVJotGvhvffOsN2fsIDfk2SHXw&hl=en&ei=CaWBSrDLCIKUtgeg_eTVCg&sa=X&oi=book_result&ct=result&resnum=1#v=onepage&q=&f=false


Sunday, August 10, 2008

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


http://creutzfeldt-jakob-disease.blogspot.com/2008/08/new-prionopathy-or-more-of-same-old-bse.html










2008

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.


http://www.cjdfoundation.org/fact.html




Friday, November 30, 2007

CJD QUESTIONNAIRE USA CWRU AND CJD FOUNDATION


http://cjdquestionnaire.blogspot.com/




Sunday, September 6, 2009

MAD COW USA 1997 SECRET VIDEO


http://madcowusda.blogspot.com/2009/09/mad-cow-usa-1997-video.html




U.S.A. HIDING MAD COW DISEASE VICTIMS AS SPORADIC CJD ? see video at bottom


http://creutzfeldt-jakob-disease.blogspot.com/2009/07/usa-hiding-mad-cow-disease-victims-as.html



DAMNING TESTIMONY FROM STANLEY PRUSINER THE NOBEL PEACE PRIZE WINNER ON PRIONS SPEAKING ABOUT ANN VENEMAN see video


http://maddeer.org/video/embedded/prusinerclip.html




2009 UPDATE ON ALABAMA AND TEXAS MAD COWS 2005 and 2006


http://bse-atypical.blogspot.com/2006/08/bse-atypical-texas-and-alabama-update.html




Office of Inspector General Semiannual Report to Congress FY 2007 - 2nd Half

Two Texas Companies Sentenced and Fined for Misbranding Meat Products In April 2007, two closely held and related Texas companies pled guilty in Federal court and were sentenced to 12 months of probation and ordered to pay $10,250 in fines for misbranding meat products. One of the companies sold adulterated meat products to a retail store in New Mexico. Additionally, portions of the invoices failed to properly and consistently identify the meat products as being from cattle more than 30 months old at time of slaughter. This information is required to be disclosed because of bovine spongiform encephalopathy (BSE, or "mad cow disease") concerns. No adulterated meat reached consumers.


http://www.usda.gov/oig/webdocs/sarc071212.pdf




Saturday, August 29, 2009

FOIA REQUEST FEED RECALL 2009 Product may have contained prohibited materials Bulk Whole Barley, Recall # V-256-2009


http://madcowfeed.blogspot.com/2009/08/foia-request-feed-recall-2009-product.html




Friday, September 4, 2009

FOIA REQUEST ON FEED RECALL PRODUCT 429,128 lbs. feed for ruminant animals may have been contaminated with prohibited material Recall # V-258-2009


http://madcowfeed.blogspot.com/2009/09/foia-request-on-feed-recall-product.html




THIS recall is not confusing ;

10,000,000+ LBS. of PROHIBITED BANNED MAD COW FEED I.E. BLOOD LACED MBM IN COMMERCE USA 2007

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

RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINES -- CLASS II

___________________________________

PRODUCT

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

CODE

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

RECALLING FIRM/MANUFACTURER

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

Firm initiated recall is ongoing.

REASON

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.

VOLUME OF PRODUCT IN COMMERCE

42,090 lbs.

DISTRIBUTION

WI

___________________________________

PRODUCT

Custom dairy premix products: MNM ALL PURPOSE Pellet, HILLSIDE/CDL Prot- Buffer Meal, LEE, M.-CLOSE UP PX Pellet, HIGH DESERT/ GHC LACT Meal, TATARKA, M CUST PROT Meal, SUNRIDGE/CDL PROTEIN Blend, LOURENZO, K PVM DAIRY Meal, DOUBLE B DAIRY/GHC LAC Mineral, WEST PIONT/GHC CLOSEUP Mineral, WEST POINT/GHC LACT Meal, JENKS, J/COMPASS PROTEIN Meal, COPPINI - 8# SPECIAL DAIRY Mix, GULICK, L-LACT Meal (Bulk), TRIPLE J - PROTEIN/LACTATION, ROCK CREEK/GHC MILK Mineral, BETTENCOURT/GHC S.SIDE MK-MN, BETTENCOURT #1/GHC MILK MINR, V&C DAIRY/GHC LACT Meal, VEENSTRA, F/GHC LACT Meal, SMUTNY, A- BYPASS ML W/SMARTA, Recall # V-025-2007

CODE

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

RECALLING FIRM/MANUFACTURER

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

REASON

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.

VOLUME OF PRODUCT IN COMMERCE

9,997,976 lbs.

DISTRIBUTION

ID and NV

END OF ENFORCEMENT REPORT FOR MARCH 21, 2007

http://www.fda.gov/bbs/topics/enforce/2007/ENF00996.html


NEW URL


http://www.fda.gov/Safety/Recalls/EnforcementReports/2007/ucm120446.htm




Thursday, March 19, 2009

MILLIONS AND MILLIONS OF POUNDS OF MAD COW FEED IN COMMERCE USA WITH ONGOING 12 YEARS OF DENIAL


http://madcowfeed.blogspot.com/2009/03/millions-and-millions-of-pounds-of-mad.html




Sunday, October 18, 2009

Wisconsin Firm Recalls Beef Tongues That Contain Prohibited Materials SRM WASHINGTON, October 17, 2009


http://madcowfeed.blogspot.com/2009/10/wisconsin-firm-recalls-beef-tongues.html




Thursday, October 15, 2009

Nebraska Firm Recalls Beef Tongues That Contain Prohibited Materials SRM WASHINGTON, Oct 15, 2009


http://madcowfeed.blogspot.com/2009/10/nebraska-firm-recalls-beef-tongues-that.html




Tuesday, July 14, 2009

U.S. Emergency Bovine Spongiform Encephalopathy Response Plan Summary and BSE Red Book Date: February 14, 2000 at 8:56 am PST

WHERE did we go wrong $$$


http://madcowtesting.blogspot.com/2009/07/us-emergency-bovine-spongiform.html




Sunday, December 28, 2008

MAD COW DISEASE USA DECEMBER 28, 2008 an 8 year review of a failed and flawed policy


http://bse-atypical.blogspot.com/2008/12/mad-cow-disease-usa-december-28-2008-8.html




Wednesday, August 20, 2008

Bovine Spongiform Encephalopathy Mad Cow Disease typical and atypical strains, was there a cover-up ? August 20, 2008


http://bse-atypical.blogspot.com/2008/08/bovine-spongiform-encephalopathy-mad.html




Monday, May 11, 2009

Rare BSE mutation raises concerns over risks to public health


http://bse-atypical.blogspot.com/2009/05/rare-bse-mutation-raises-concerns-over.html




Tuesday, July 14, 2009

U.S. Emergency Bovine Spongiform Encephalopathy Response Plan Summary and BSE Red Book Date: February 14, 2000 at 8:56 am PST

WHERE did we go wrong $$$


http://madcowtesting.blogspot.com/2009/07/us-emergency-bovine-spongiform.html




Sunday, June 07, 2009

L-TYPE-BSE, H-TYPE-BSE, C-TYPE-BSE, IBNC-TYPE-BSE, TME, CWD, SCRAPIE, CJD, NORTH AMERICA


http://bse-atypical.blogspot.com/2009/06/l-type-bse-h-type-bse-c-type-bse-ibnc.html




Sunday, May 10, 2009

Identification and characterization of bovine spongiform encephalopathy cases diagnosed and NOT diagnosed in the United States


http://bse-atypical.blogspot.com/2009/05/identification-and-characterization-of.html




Docket APHIS-2006-0026 Docket Title Bovine Spongiform Encephalopathy; Animal Identification and Importation of Commodities Docket Type Rulemaking Document APHIS-2006-0026-0001 Document Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions, Identification of Ruminants and Processing and Importation of Commodities Public Submission APHIS-2006-0026-0012 Public Submission Title Comment from Terry S Singletary


http://www.regulations.gov/fdmspublic/component/main?main=DocumentDetail&o=09000064801e47e1




Docket APHIS-2006-0041 Docket Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived from Bovines Commodities Docket Type Rulemaking Document APHIS-2006-0041-0001 Document Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived From Bovines Public Submission APHIS-2006-0041-0028 Public Submission Title Comment from Terry S Singletary

Comment 2006-2007 USA AND OIE POISONING GLOBE WITH BSE MRR POLICY

THE USA is in a most unique situation, one of unknown circumstances with human and animal TSE. THE USA has the most documented TSE in different species to date, with substrains growing in those species (BSE/BASE in cattle and CWD in deer and elk, there is evidence here with different strains), and we know that sheep scrapie has over 20 strains of the typical scrapie with atypical scrapie documented and also BSE is very likely to have passed to sheep. all of which have been rendered and fed back to animals for human and animal consumption, a frightening scenario. WE do not know the outcome, and to play with human life around the globe with the very likely TSE tainted products from the USA, in my opinion is like playing Russian roulette, of long duration, with potential long and enduring consequences, of which once done, cannot be undone. These are the facts as I have come to know through daily and extensive research of TSE over 9 years, since 12/14/97. I do not pretend to have all the answers, but i do know to continue to believe in the ukbsenvcjd only theory of transmission to humans of only this one strain from only this one TSE from only this one part of the globe, will only lead to further failures, and needless exposure to humans from all strains of TSE, and possibly many more needless deaths from TSE via a multitude of proven routes and sources via many studies with primates and rodents and other species.

MY personal belief, since you ask, is that not only the Canadian border, but the USA border, and the Mexican border should be sealed up tighter than a drum for exporting there TSE tainted products, until a validated, 100% sensitive test is available, and all animals for human and animal consumption are tested. all we are doing is the exact same thing the UK did with there mad cow poisoning when they exported it all over the globe, all the while knowing what they were doing. this BSE MRR policy is nothing more than a legal tool to do just exactly what the UK did, thanks to the OIE and GW, it's legal now. and they executed Saddam for poisoning ???

go figure. ...


http://www.regulations.gov/fdmspublic/component/main?main=DocumentDetail&o=09000064801f8151




Docket APHIS-2006-0041 Docket Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived from Bovines Commodities Docket Type Rulemaking Document APHIS-2006-0041-0001 Document Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived From Bovines Public Submission APHIS-2006-0041-0028.1 Public Submission Title Attachment to Singletary comment

January 28, 2007

Greetings APHIS,

I would kindly like to submit the following to ;

BSE; MRR; IMPORTATION OF LIVE BOVINES AND PRODUCTS DERIVED FROM BOVINES [Docket No. APHIS-2006-0041] RIN 0579-AC01


http://www.regulations.gov/fdmspublic/ContentViewer?objectId=09000064801f8152&disposition=attachment&contentType=msw8




Thursday, October 15, 2009

CVM Annual Report Fiscal Year 2008: October 1, 2007-September 30, 2008 (BSE)


http://madcowusda.blogspot.com/2009/10/cvm-annual-report-fiscal-year-2008.html




Wednesday, August 19, 2009

CFIA Enhances Animal Disease Reporting


http://madcowtesting.blogspot.com/2009/08/cfia-enhances-animal-disease-reporting.html




Thursday, October 15, 2009

SCRAPIE UPDATE CANADA 2009 (typical and atypical cases)


http://scrapie-usa.blogspot.com/2009/10/scrapie-update-canada-2009-typical-and.html




SEAC OCTOBER 2009

• Are some commoner types of neurodegenerative disease (including Alzheimer's disease and Parkinson's disease) also transmissible? Some recent scientific research has suggested this possibility


http://www.seac.gov.uk/pdf/hol-response091008.pdf




Alzheimer's and CJD


http://betaamyloidcjd.blogspot.com/




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

SEAC Science and Technology Committee's investigation of research funding priorities on behalf of the Advisory Committee on Dangerous Pathogens TSE

8 October 2009

1

House of Lords Science and Technology Committee

Setting science and technology research funding priorities Evidence from the UK Government’s Spongiform Encephalopathy Advisory Committees:

1. Advisory Committee on Dangerous Pathogens Transmissible Spongiform Encephalopathy Working Group (ACDP TSE WG),

2. CJD Incidents Panel,

3. Engineering and Science Advisory Committee into the decontamination of surgical instruments including Prion Removal (ESAC Pr)

4. Spongiform Encephalopathy Advisory Committee (SEAC)

1. The handling of transmissible spongiform encephalopathies (TSEs) has important lessons for the work that the House of Lords Science and Technology Committee intends to focus on, specifically:

• How decisions are made to fund research to meet societal needs

• The balance of funding for targeted versus unsolicited response-mode curiosity-driven research, and

• How research is commissioned in Government departments and agencies

2. The transmissible spongiform encephalopathies (TSEs), also known as prion diseases, are a group of fatal transmissible neurodegenerative disorders of man and animals, characterized by the “spongy” microscopic appearance of the brain in affected animals and by a link with a ubiquitous protein, the prion protein (PrP), a misfolded form of which is widely believed (though never unequivocally proved) to be the infectious agent or prion. The most widely known examples of these diseases are bovine spongiform encephalopathy (BSE) in cattle, scrapie in sheep and goats and Creutzfeldt-Jakob disease (CJD) in humans. BSE emerged in the UK in the 1980s, has been reported in many other countries and is responsible for the human disease, variant CJD (vCJD).

3. TSEs are thus a group of diseases that, in a short space of time, gave rise to significant health effects in animals and humans and resulted in massive economic loss. In identifying the solutions that were required, science and scientific research, both basic and applied, were critical. The recent history of the diseases and the research into them therefore merits careful consideration by the House of Lords Science and Technology Committee, as many of the specific aspects of that history are relevant to the general principles that the Committee is looking into.

4. SEAC is the government’s overarching committee for advising on the science of TSEs and thereby assessing risk to the public. ACDP TSE WG, CJDIP and ESAC Pr are in their different ways involved in developing practical advice to reduce the spread of TSEs (risk management), most notably spread of CJD/vCJD from person to person via contaminated surgical instruments or via blood transfusion.

2

What is the overall objective of publicly-funded science and technology research?

5. The overall objective of publicly-funded science and technology research, the public good, has to be considered in a broader and longer term context than that of policy, as the latter is often understood within government. This policy, in practice, is, inevitably, strongly influenced by electoral and media cycles whereas effective scientific policy has to be constructed around a much longer term administrative cycle. This is particularly aptly illustrated by TSEs, in which the slow progression of the diseases can make developing reliable answers to scientific questions, necessarily, a long term undertaking.

6. Publicly funded TSE research in the UK was stimulated by substantial public funding in the late 1980s/ early 1990s, initially in response to the threat that the emerging BSE epidemic in cattle posed to animal health and later following the recognition, in 1996, that BSE was linked to vCJD and posed a public health risk.. The need to limit the damage to health and wellbeing from BSE gave rise to a number of intensely practical questions such as the nature of the infection and the infectious agent, the distribution of the agent in different animal species and whether barriers to transmission existed between certain species. These questions could not be answered without an investment in basic science. Equally some of the key techniques for characterising the diseases, such as biological and molecular strain typing of the responsible agents had been developed in the 1970s and 1980s to distinguish different isolates of scrapie. This latter work had taken place at a time when the policy community put so little emphasis on TSEs that these developments were seen, at the time, as of little practical application. How are science and technology research priorities co-ordinated across government and between government and the relevant funding organizations? Who is responsible for ensuring that research gaps are filled?

7. Balancing curiosity driven research with research driven by departmental and policy needs requires that both research councils and the departments themselves are in a position to commission meaningful research and that this research can be effectively co-ordinated.

8. The model by which research funding priorities has been co-ordinated between government departments and the research councils has been the TSE Joint Funders Advisory Group which has been sufficiently successful to be emulated for novel H1N1 influenza A virus (“swine flu”) research. However, the decline of the BSE and vCJD epidemics has led to a recent disinvestment in the field. This is premature. Those of us who are members of the three risk-management committees, particularly, are aware of questions that, if answered, would facilitate disease control, by allowing potentially the relaxation of certain expensive contemporary control measures as well as those answers having implications for other more widespread diseases. There are equally other widespread TSEs such as Chronic Wasting Disease (CWD) of mule deer and elk in the United States and Canada that may yet show the potential to infect man, as well as newly identified TSEs such as bovine amyloid spongiform encephalopathy (BASE), and atypical scrapie, whose potential to be a human

3

health problem is not yet fully understood. In humans there is a recently discovered ‘new’ TSE called protease sensitive prionopathy (PSP), the significance of which is, as yet, unclear.

9. More specific examples of unanswered questions with health implications are:

• Will the eventual elimination of classical scrapie in the EU leave an ecological niche for other TSEs such as BSE or atypical scrapie?

• Is CWD transmissible to humans?

• Can a reliable ante mortem diagnostic blood test for vCJD be developed?

• What is the true prevalence of v CJD infection (as opposed to overt disease) in the UK?

• Are some commoner types of neurodegenerative disease (including Alzheimer's disease and Parkinson's disease) also transmissible? Some recent scientific research has suggested this possibility

• Could cases of protease sensitive prionopathy (PSP) be missed by conventional tests which, in all other TSEs, rely on the resistance of the prion protein in the nervous system that accompanies disease to digestion by protease enzymes?

• Can we develop reliable methods for removing and detecting protein on re-usable surgical instruments?

10. These are frequently highly practical questions impacting on very expensive policy options but needing to be informed by scientific work that may more appropriately be described as basic rather than applied. The cost of funding such work could well be trivial compared to the precautionary measures that are currently being put in place to mitigate such possible but unproven risks. To what extent should publicly-funded science and technology research be focussed on areas of potential economic importance? How should these areas be identified?

11. We would argue that the evaluation of the economic importance of science and technology research needs to be based on a model that is sufficiently sophisticated to acknowledge adequately longer term economic benefit. For example, DEFRA have developed a prioritization tool which ranks animal diseases according to a number of variables – for example impact on public health, animal welfare, international trade and wider society.

12. Thus any method for evaluating the economic importance of research should be able to recognize that TSE research in the UK remains a vibrant field in which there are a number of young researchers making real progress with implications for a variety of diseases and disease processes. The TSEs themselves retain the capacity to surprise and although BSE and vCJD appear to be declining, other

4

health questions that give rise to circumstances that lead to human illness, economic loss and political embarrassment, seem likely. Further, we have now also arrived at the point where a research infrastructure (laboratories, animal facilities, cell lines, animal lines, reagents, trained personnel) with strong international links within the European Union and to Japan and North America, has been established in the UK that can allow complicated questions to be answered efficiently. There would be a considerable opportunity cost to losing this resource.

24 September 2009

The Secretariat on behalf of:

• Advisory Committee on Dangerous Pathogens Transmissible Spongiform Encephalopathy Working Group (ACDP TSE WG),

• CJD Incidents Panel,

• Engineering and Science Advisory Committee into the decontamination of surgical instruments including Prion Removal (ESAC Pr)

• Spongiform Encephalopathy Advisory Committee (SEAC)


http://www.seac.gov.uk/pdf/hol-response091008.pdf




Sunday, May 17, 2009

WHO WILL WATCH THE CHILDREN ? SCHOOL LUNCH PROGRAM FROM DOWNER CATTLE UPDATE


http://downercattle.blogspot.com/2009/05/who-will-watch-children.html




http://downercattle.blogspot.com/




Sunday, September 6, 2009

MAD COW USA 1997 SECRET VIDEO


http://madcowusda.blogspot.com/2009/09/mad-cow-usa-1997-video.html




U.S.A. HIDING MAD COW DISEASE VICTIMS AS SPORADIC CJD ? see video at bottom


http://creutzfeldt-jakob-disease.blogspot.com/2009/07/usa-hiding-mad-cow-disease-victims-as.html




DAMNING TESTIMONY FROM STANLEY PRUSINER THE NOBEL PEACE PRIZE WINNER ON PRIONS SPEAKING ABOUT ANN VENEMAN
SEE VIDEO


http://maddeer.org/video/embedded/prusinerclip.html





2009 UPDATE ON ALABAMA AND TEXAS MAD COWS 2005 and 2006


http://bse-atypical.blogspot.com/2006/08/bse-atypical-texas-and-alabama-update.html




http://creutzfeldt-jakob-disease.blogspot.com/2009/08/characteristics-of-established-and.html




Office of Inspector General Semiannual Report to Congress FY 2007 - 2nd Half

Two Texas Companies Sentenced and Fined for Misbranding Meat Products In April 2007, two closely held and related Texas companies pled guilty in Federal court and were sentenced to 12 months of probation and ordered to pay $10,250 in fines for misbranding meat products. One of the companies sold adulterated meat products to a retail store in New Mexico. Additionally, portions of the invoices failed to properly and consistently identify the meat products as being from cattle more than 30 months old at time of slaughter. This information is required to be disclosed because of bovine spongiform encephalopathy (BSE, or "mad cow disease") concerns. No adulterated meat reached consumers.


http://www.usda.gov/oig/webdocs/sarc071212.pdf




Tuesday, July 14, 2009

U.S. Emergency Bovine Spongiform Encephalopathy Response Plan Summary and BSE Red Book Date: February 14, 2000 at 8:56 am PST

WHERE did we go wrong $$$


http://madcowtesting.blogspot.com/2009/07/us-emergency-bovine-spongiform.html




Sunday, December 28, 2008

MAD COW DISEASE USA DECEMBER 28, 2008 an 8 year review of a failed and flawed policy


http://bse-atypical.blogspot.com/2008/12/mad-cow-disease-usa-december-28-2008-8.html




Wednesday, August 20, 2008

Bovine Spongiform Encephalopathy Mad Cow Disease typical and atypical strains, was there a cover-up ? August 20, 2008


http://bse-atypical.blogspot.com/2008/08/bovine-spongiform-encephalopathy-mad.html




Saturday, June 13, 2009

Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States 2003 revisited 2009


http://cjdusa.blogspot.com/2009/06/monitoring-occurrence-of-emerging-forms.html




Tuesday, August 11, 2009

Characteristics of Established and Proposed Sporadic Creutzfeldt-Jakob Disease Variants

Brian S. Appleby, MD; Kristin K. Appleby, MD; Barbara J. Crain, MD, PhD; Chiadi U. Onyike, MD, MHS; Mitchell T. Wallin, MD, MPH; Peter V. Rabins, MD, MPH

Background: The classic Creutzfeldt-Jakob disease (CJD), Heidenhain, and Oppenheimer-Brownell variants are sporadic CJD (sCJD) phenotypes frequently described in the literature, but many cases present with neuropsychiatric symptoms, suggesting that there may be additional sCJD phenotypes.

Objective: To characterize clinical, diagnostic, and molecular features of 5 sCJD variants.

Design: Retrospective analysis.

Setting: The Johns Hopkins and Veterans Administration health care systems.

Participants: Eighty-eight patients with definite or probable sCJD.

Main Outcome Measures: Differences in age at onset, illness progression, diagnostic test results, and molecular subtype.

Results: The age at onset differed among sCJD variants (P=.03); the affective variant had the youngest mean age at onset (59.7 years). Survival time (P
.001) and the time to clinical presentation (P=.003) differed among groups. Patients with the classic CJD phenotype had the shortest median survival time from symptom onset (66 days) and those who met criteria for the affective sCJD variant had the longest (421 days) and presented to clinicians significantly later (median time from onset to presentation, 92 days; P=.004). Cerebrospinal fluid analyses were positive for 14-3-3 protein in all of the affective variants, regardless of illness duration. Periodic sharp-wave complexes were not detected on any of the electroencephalography tracings in the Oppenheimer-Brownell group; basal ganglia hyperintensity was not detected on brain magnetic resonance imaging in this group either. All of the Heidenhain variants were of the methionine/ methionine type 1 molecular subtype.

Conclusions: The classic CJD phenotype and the Heidenhain, Oppenheimer-Brownell, cognitive, and affective sCJD variants differ by age at disease onset, survival time, and diagnostic test results. Characteristics of these 5 phenotypes are provided to facilitate further clinicopathologic investigation that may lead to more reliable and timely diagnoses of sCJD.

Arch Neurol. 2009;66(2):208-215

snip...

COMMENT

snip...see full text ;


http://creutzfeldt-jakob-disease.blogspot.com/2009/08/characteristics-of-established-and.html




Office of Inspector General Semiannual Report to Congress FY 2007 - 2nd Half

Two Texas Companies Sentenced and Fined for Misbranding Meat Products In April 2007, two closely held and related Texas companies pled guilty in Federal court and were sentenced to 12 months of probation and ordered to pay $10,250 in fines for misbranding meat products. One of the companies sold adulterated meat products to a retail store in New Mexico. Additionally, portions of the invoices failed to properly and consistently identify the meat products as being from cattle more than 30 months old at time of slaughter. This information is required to be disclosed because of bovine spongiform encephalopathy (BSE, or "mad cow disease") concerns. No adulterated meat reached consumers.


http://www.usda.gov/oig/webdocs/sarc071212.pdf




Saturday, August 29, 2009

FOIA REQUEST FEED RECALL 2009 Product may have contained prohibited materials Bulk Whole Barley, Recall # V-256-2009


http://madcowfeed.blogspot.com/2009/08/foia-request-feed-recall-2009-product.html




Friday, September 4, 2009

FOIA REQUEST ON FEED RECALL PRODUCT 429,128 lbs. feed for ruminant animals may have been contaminated with prohibited material Recall # V-258-2009


http://madcowfeed.blogspot.com/2009/09/foia-request-on-feed-recall-product.html




THIS recall is not confusing ;

10,000,000+ LBS. of PROHIBITED BANNED MAD COW FEED I.E. BLOOD LACED MBM IN COMMERCE USA 2007

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

RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINES -- CLASS II

___________________________________

PRODUCT

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

CODE

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

RECALLING FIRM/MANUFACTURER

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

Firm initiated recall is ongoing.

REASON

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.

VOLUME OF PRODUCT IN COMMERCE

42,090 lbs.

DISTRIBUTION

WI

___________________________________

PRODUCT

Custom dairy premix products: MNM ALL PURPOSE Pellet, HILLSIDE/CDL Prot- Buffer Meal, LEE, M.-CLOSE UP PX Pellet, HIGH DESERT/ GHC LACT Meal, TATARKA, M CUST PROT Meal, SUNRIDGE/CDL PROTEIN Blend, LOURENZO, K PVM DAIRY Meal, DOUBLE B DAIRY/GHC LAC Mineral, WEST PIONT/GHC CLOSEUP Mineral, WEST POINT/GHC LACT Meal, JENKS, J/COMPASS PROTEIN Meal, COPPINI - 8# SPECIAL DAIRY Mix, GULICK, L-LACT Meal (Bulk), TRIPLE J - PROTEIN/LACTATION, ROCK CREEK/GHC MILK Mineral, BETTENCOURT/GHC S.SIDE MK-MN, BETTENCOURT #1/GHC MILK MINR, V&C DAIRY/GHC LACT Meal, VEENSTRA, F/GHC LACT Meal, SMUTNY, A- BYPASS ML W/SMARTA, Recall # V-025-2007

CODE

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

RECALLING FIRM/MANUFACTURER

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

REASON

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.

VOLUME OF PRODUCT IN COMMERCE

9,997,976 lbs.

DISTRIBUTION

ID and NV

END OF ENFORCEMENT REPORT FOR MARCH 21, 2007

http://www.fda.gov/bbs/topics/enforce/2007/ENF00996.html


NEW URL


http://www.fda.gov/Safety/Recalls/EnforcementReports/2007/ucm120446.htm




Thursday, March 19, 2009

MILLIONS AND MILLIONS OF POUNDS OF MAD COW FEED IN COMMERCE USA WITH ONGOING 12 YEARS OF DENIAL


http://madcowfeed.blogspot.com/2009/03/millions-and-millions-of-pounds-of-mad.html






Monday, May 11, 2009

Rare BSE mutation raises concerns over risks to public health


http://bse-atypical.blogspot.com/2009/05/rare-bse-mutation-raises-concerns-over.html





Sunday, June 07, 2009

L-TYPE-BSE, H-TYPE-BSE, C-TYPE-BSE, IBNC-TYPE-BSE, TME, CWD, SCRAPIE, CJD, NORTH AMERICA


http://bse-atypical.blogspot.com/2009/06/l-type-bse-h-type-bse-c-type-bse-ibnc.html




Sunday, May 10, 2009

Identification and characterization of bovine spongiform encephalopathy cases diagnosed and NOT diagnosed in the United States


http://bse-atypical.blogspot.com/2009/05/identification-and-characterization-of.html




Docket APHIS-2006-0026 Docket Title Bovine Spongiform Encephalopathy; Animal Identification and Importation of Commodities Docket Type Rulemaking Document APHIS-2006-0026-0001 Document Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions, Identification of Ruminants and Processing and Importation of Commodities Public Submission APHIS-2006-0026-0012 Public Submission Title Comment from Terry S Singletary


http://www.regulations.gov/fdmspublic/component/main?main=DocumentDetail&o=09000064801e47e1




Docket APHIS-2006-0041 Docket Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived from Bovines Commodities Docket Type Rulemaking Document APHIS-2006-0041-0001 Document Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived From Bovines Public Submission APHIS-2006-0041-0028 Public Submission Title Comment from Terry S Singletary

Comment 2006-2007 USA AND OIE POISONING GLOBE WITH BSE MRR POLICY

THE USA is in a most unique situation, one of unknown circumstances with human and animal TSE. THE USA has the most documented TSE in different species to date, with substrains growing in those species (BSE/BASE in cattle and CWD in deer and elk, there is evidence here with different strains), and we know that sheep scrapie has over 20 strains of the typical scrapie with atypical scrapie documented and also BSE is very likely to have passed to sheep. all of which have been rendered and fed back to animals for human and animal consumption, a frightening scenario. WE do not know the outcome, and to play with human life around the globe with the very likely TSE tainted products from the USA, in my opinion is like playing Russian roulette, of long duration, with potential long and enduring consequences, of which once done, cannot be undone. These are the facts as I have come to know through daily and extensive research of TSE over 9 years, since 12/14/97. I do not pretend to have all the answers, but i do know to continue to believe in the ukbsenvcjd only theory of transmission to humans of only this one strain from only this one TSE from only this one part of the globe, will only lead to further failures, and needless exposure to humans from all strains of TSE, and possibly many more needless deaths from TSE via a multitude of proven routes and sources via many studies with primates and rodents and other species.

MY personal belief, since you ask, is that not only the Canadian border, but the USA border, and the Mexican border should be sealed up tighter than a drum for exporting there TSE tainted products, until a validated, 100% sensitive test is available, and all animals for human and animal consumption are tested. all we are doing is the exact same thing the UK did with there mad cow poisoning when they exported it all over the globe, all the while knowing what they were doing. this BSE MRR policy is nothing more than a legal tool to do just exactly what the UK did, thanks to the OIE and GW, it's legal now. and they executed Saddam for poisoning ???

go figure. ...


http://www.regulations.gov/fdmspublic/component/main?main=DocumentDetail&o=09000064801f8151




Docket APHIS-2006-0041 Docket Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived from Bovines Commodities Docket Type Rulemaking Document APHIS-2006-0041-0001 Document Title Bovine Spongiform Encephalopathy; Minimal-Risk Regions; Importation of Live Bovines and Products Derived From Bovines Public Submission APHIS-2006-0041-0028.1 Public Submission Title Attachment to Singletary comment

January 28, 2007

Greetings APHIS,

I would kindly like to submit the following to ;

BSE; MRR; IMPORTATION OF LIVE BOVINES AND PRODUCTS DERIVED FROM BOVINES [Docket No. APHIS-2006-0041] RIN 0579-AC01


http://www.regulations.gov/fdmspublic/ContentViewer?objectId=09000064801f8152&disposition=attachment&contentType=msw8




Sunday, August 10, 2008

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


http://creutzfeldt-jakob-disease.blogspot.com/2008/08/new-prionopathy-or-more-of-same-old-bse.html




SEAC OCTOBER 2009

• Are some commoner types of neurodegenerative disease (including Alzheimer's disease and Parkinson's disease) also transmissible? Some recent scientific research has suggested this possibility


http://www.seac.gov.uk/pdf/hol-response091008.pdf




Thursday, February 26, 2009

'Harmless' prion protein linked to Alzheimer's disease Non-infectious form of prion protein could cause brain degeneration ???


http://betaamyloidcjd.blogspot.com/2009/02/harmless-prion-protein-linked-to.html




CJD1/9 0185

Ref: 1M51A

IN STRICT CONFIDENCE

TRANSMISSION OF ALZHEIMER-TYPE PLAQUES TO PRIMATES

1. CMO will wish to be aware that a meeting was held at DH yesterday, 4 January, to discuss the above findings. It was chaired by Professor Murray (Chairman of the MRC Co-ordinating Committee on Research in the Spongiform Encephalopathies in Man), and attended by relevant experts in the fields of Neurology, Neuropathology, molecular biology, amyloid biochemistry, and the spongiform encephalopathies, and by representatives of the MRC and AFRC.

2. Briefly, the meeting agreed that:

i) Dr Ridley et als findings of experimental induction of p amyloid in primates were valid, interesting and a significant advance in the understanding of neurodegeneradve disorders;

ii) there were no immediate implications for the public health, and no further safeguards were thought to be necessary at present; and

iii) additional research was desirable, both epidemiological and at the molecular level. Possible avenues are being followed up by DH and the MRC, but the details will require further discussion.

93/01.05/4.1tss


http://www.bseinquiry.gov.uk/files/yb/1993/01/05004001.pdf




Regarding Alzheimer's disease

(note the substantial increase on a yearly basis)


http://www.bseinquiry.gov.uk/files/yb/1988/07/08014001.pdf




snip...

The pathogenesis of these diseases was compared to Alzheimer's disease at a molecular level...

snip...


http://www.bseinquiry.gov.uk/files/yb/1990/03/12003001.pdf




And NONE of this is relevant to BSE?

There is also the matter whether the spectrum of ''prion disease'' is wider than that recognized at present.


http://www.bseinquiry.gov.uk/files/yb/1990/07/06005001.pdf




http://www.bseinquiry.gov.uk/files/yb/1990/07/09001001.pdf





BSE101/1 0136

IN CONFIDENCE

5 NOV 1992

CMO From: Dr J S Metters DCMO 4 November 1992

TRANSMISSION OF ALZHEIMER TYPE PLAQUES TO PRIMATES


http://www.bseinquiry.gov.uk/files/yb/1992/11/04001001.pdf




also, see the increase of Alzheimer's from 1981 to 1986


http://www.bseinquiry.gov.uk/files/yb/1988/07/08014001.pdf




Tuesday, August 26, 2008

Alzheimer's Transmission of AA-amyloidosis: Similarities with Prion Disorders NEUROPRION 2007 FC4.3


http://betaamyloidcjd.blogspot.com/2008/08/alzheimers-transmission-of-aa.html




see full text ;


http://betaamyloidcjd.blogspot.com/2009/02/harmless-prion-protein-linked-to.html




Alzheimer's and CJD


http://betaamyloidcjd.blogspot.com/




Saturday, March 22, 2008

10 Million Baby Boomers to have Alzheimer's in the coming decades 2008 Alzheimer's disease facts and figures


http://betaamyloidcjd.blogspot.com/2008/03/association-between-deposition-of-beta.html




re-Association between Deposition of Beta-Amyloid and Pathological Prion Protein in Sporadic Creutzfeldt-Jakob Disease


http://betaamyloidcjd.blogspot.com/2008/04/re-association-between-deposition-of.html




Thursday, December 04, 2008 2:37 PM

"we have found that H-BSE can infect humans."

personal communication with Professor Kong. ...TSS

see full text ;


http://bse-atypical.blogspot.com/2009/02/atypical-bse-north-america-update.html




PR-26

NOR98 SHOWS MOLECULAR FEATURES REMINISCENT OF GSS

R. Nonno1, E. Esposito1, G. Vaccari1, E. Bandino2, M. Conte1, B. Chiappini1, S. Marcon1, M. Di Bari1, S.L. Benestad3, U. Agrimi1 1 Istituto Superiore di Sanità, Department of Food Safety and Veterinary Public Health, Rome, Italy (romolo.nonno@iss.it); 2 Istituto Zooprofilattico della Sardegna, Sassari, Italy; 3 National Veterinary Institute, Department of Pathology, Oslo, Norway

Molecular variants of PrPSc are being increasingly investigated in sheep scrapie and are generally referred to as “atypical” scrapie, as opposed to “classical scrapie”. Among the atypical group, Nor98 seems to be the best identified. We studied the molecular properties of Italian and Norwegian Nor98 samples by WB analysis of brain homogenates, either untreated, digested with different concentrations of proteinase K, or subjected to enzymatic deglycosylation. The identity of PrP fragments was inferred by means of antibodies spanning the full PrP sequence. We found that undigested brain homogenates contain a Nor98-specific PrP fragment migrating at 11 kDa (PrP11), truncated at both the C-terminus and the N-terminus, and not N-glycosylated. After mild PK digestion, Nor98 displayed full-length PrP (FL-PrP) and N-glycosylated C-terminal fragments (CTF), along with increased levels of PrP11. Proteinase K digestion curves (0,006-6,4 mg/ml) showed that FL-PrP and CTF are mainly digested above 0,01 mg/ml, while PrP11 is not entirely digested even at the highest concentrations, similarly to PrP27-30 associated with classical scrapie. Above 0,2 mg/ml PK, most Nor98 samples showed only PrP11 and a fragment of 17 kDa with the same properties of PrP11, that was tentatively identified as a dimer of PrP11. Detergent solubility studies showed that PrP11 is insoluble in 2% sodium laurylsorcosine and is mainly produced from detergentsoluble, full-length PrPSc. Furthermore, among Italian scrapie isolates, we found that a sample with molecular and pathological properties consistent with Nor98 showed plaque-like deposits of PrPSc in the thalamus when the brain was analysed by PrPSc immunohistochemistry. Taken together, our results show that the distinctive pathological feature of Nor98 is a PrP fragment spanning amino acids ~ 90-155. This fragment is produced by successive N-terminal and C-terminal cleavages from a full-length and largely detergent-soluble PrPSc, is produced in vivo and is extremely resistant to PK digestion. Intriguingly, these conclusions suggest that some pathological features of Nor98 are reminiscent of Gerstmann-Sträussler-Scheinker disease.

119


http://www.neuroprion.com/pdf_docs/conferences/prion2006/abstract_book.pdf




When Atypical Scrapie cross species barriers

Authors

Andreoletti O., Herva M. H., Cassard H., Espinosa J. C., Lacroux C., Simon S., Padilla D., Benestad S. L., Lantier F., Schelcher F., Grassi J., Torres, J. M., UMR INRA ENVT 1225, Ecole Nationale Veterinaire de Toulouse.France; ICISA-INlA, Madrid, Spain; CEA, IBiTec-5, DSV, CEA/Saclay, Gif sur Yvette cedex, France; National Veterinary Institute, Postboks 750 Sentrum, 0106 Oslo, Norway, INRA IASP, Centre INRA de Tours, 3738O Nouzilly, France.

Content

Atypical scrapie is a TSE occurring in small ruminants and harbouring peculiar clinical, epidemiological and biochemical properties. Currently this form of disease is identified in a large number of countries. In this study we report the transmission of an atypical scrapie isolate through different species barriers as modeled by transgenic mice (Tg) expressing different species PRP sequence.

The donor isolate was collected in 1995 in a French commercial sheep flock. inoculation into AHQ/AHQ sheep induced a disease which had all neuro-pathological and biochemical characteristics of atypical scrapie. Transmitted into Transgenic mice expressing either ovine or PrPc, the isolate retained all the described characteristics of atypical scrapie.

Surprisingly the TSE agent characteristics were dramatically different v/hen passaged into Tg bovine mice. The recovered TSE agent had biological and biochemical characteristics similar to those of atypical BSE L in the same mouse model. Moreover, whereas no other TSE agent than BSE were shown to transmit into Tg porcine mice, atypical scrapie was able to develop into this model, albeit with low attack rate on first passage.

Furthermore, after adaptation in the porcine mouse model this prion showed similar biological and biochemical characteristics than BSE adapted to this porcine mouse model. Altogether these data indicate.

(i) the unsuspected potential abilities of atypical scrapie to cross species barriers

(ii) the possible capacity of this agent to acquire new characteristics when crossing species barrier

These findings raise some interrogation on the concept of TSE strain and on the origin of the diversity of the TSE agents and could have consequences on field TSE control measures.


http://www.neuroprion.org/resources/pdf_docs/conferences/prion2008/abstract-book-prion2008.pdf




P03.141

Aspects of the Cerebellar Neuropathology in Nor98

Gavier-Widén, D1; Benestad, SL2; Ottander, L1; Westergren, E1 1National Veterinary Insitute, Sweden; 2National Veterinary Institute,

Norway Nor98 is a prion disease of old sheep and goats. This atypical form of scrapie was first described in Norway in 1998. Several features of Nor98 were shown to be different from classical scrapie including the distribution of disease associated prion protein (PrPd) accumulation in the brain. The cerebellum is generally the most affected brain area in Nor98. The study here presented aimed at adding information on the neuropathology in the cerebellum of Nor98 naturally affected sheep of various genotypes in Sweden and Norway. A panel of histochemical and immunohistochemical (IHC) stainings such as IHC for PrPd, synaptophysin, glial fibrillary acidic protein, amyloid, and cell markers for phagocytic cells were conducted. The type of histological lesions and tissue reactions were evaluated. The types of PrPd deposition were characterized. The cerebellar cortex was regularly affected, even though there was a variation in the severity of the lesions from case to case. Neuropil vacuolation was more marked in the molecular layer, but affected also the granular cell layer. There was a loss of granule cells. Punctate deposition of PrPd was characteristic. It was morphologically and in distribution identical with that of synaptophysin, suggesting that PrPd accumulates in the synaptic structures. PrPd was also observed in the granule cell layer and in the white matter. The pathology features of Nor98 in the cerebellum of the affected sheep showed similarities with those of sporadic Creutzfeldt-Jakob disease in humans.

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


http://www.prion2007.com/pdf/Prion%20Book%20of%20Abstracts.pdf




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)


http://www.pnas.org/cgi/content/abstract/0502296102v1




NOR-98 ATYPICAL SCRAPIE 5 cases documented in USA in 5 different states USA 2007


http://nor-98.blogspot.com/2008/04/seac-spongiform-encephalopathy-advisory.html




Tuesday, June 3, 2008 SCRAPIE USA UPDATE JUNE 2008 NOR-98 REPORTED PA


http://nor-98.blogspot.com/2008/06/scrapie-usa-update-june-2008-nor-98.html




http://nor-98.blogspot.com/




Monday, September 1, 2008

RE-FOIA OF DECLARATION OF EXTRAORDINARY EMERGENCY BECAUSE OF AN ATYPICAL T.S.E. (PRION DISEASE) OF FOREIGN ORIGIN IN THE UNITED STATES [No. 00-072-1]

September 1, 2008


http://foiamadsheepmadrivervalley.blogspot.com/2008/09/re-foia-of-declaration-of-extraordinary.html




Monday, December 1, 2008

When Atypical Scrapie cross species barriers


http://nor-98.blogspot.com/2008/12/when-atypical-scrapie-cross-species.html



Tuesday, August 04, 2009

Susceptibilities of Nonhuman Primates to Chronic Wasting Disease


http://chronic-wasting-disease.blogspot.com/2009/08/susceptibilities-of-nonhuman-primates.html




Terry S. Singeltary Sr.

P.O. Box 42

Bacliff, Texas USA 77518

Scientific Opinion on BSE Risk in Bovine Intestines Question number: EFSA-Q-2009-00226

Scientific Opinion on BSE Risk in Bovine Intestines Question number: EFSA-Q-2009-00226

Adopted: 10 September 2009 Summary (0.1Mb)

Opinion (0.1Mb)

Summary

Following a request from the European Commission (EC), the Panel on Biological Hazards (BIOHAZ) was asked to deliver a scientific opinion on the BSE related risk of bovine intestines used for casings. Regulation (EC) No 999/2001 of the European Parliament and of the Council stipulates that certain tissues from bovine, ovine and caprine animals must be considered as Specified Risk Material (SRM) and must be removed from the food and feed chain to protect the health of consumers against the risk of bovine transmissible spongiform encephalopathies (BSE). The intestines, from the duodenum to the rectum, of bovine animals of all ages are currently included in the list of SRM. The “TSE roadmap” prepared by the EC details the short, middle and long term actions on TSE measures such as SRM removal and sets the objectives to ensure and maintain the existing high level of consumer protection. It allows for amendments of the current SRM list based on new evolving scientific knowledge while ensuring and maintaining a high level of consumer protection.

Specifically, the mandate asked the BIOHAZ panel to evaluate the scientific validity of a report prepared by Det Norske Veritas Ltd" (DNV) for the Swiss Cervelas task force. This report provides an assessment of the current potential human exposure to BSE infectivity that could result from eating sausages made with EU bovine casings. The BIOHAZ panel was further requested to evaluate the conclusions of the DNV report and, if it was considered necessary based on the report and any other new relevant scientific information, to provide a re-assessment of the BSE related risk of bovine intestines after processing into natural sausage casings.

The BIOHAZ panel evaluated the risk assessment as described in the DNV report, and took into account the relevant previous EFSA opinions as well as new scientific data on the same subject.

New but limited experimental scientific data demonstrate that in addition to ileum, also jejunum may harbour infectivity when a large BSE inoculum dose was used to experimentally infect cattle. With regard to the DNV Report, the BIOHAZ Panel considers its approach (concept and methodology) scientifically sound, whereas the interpretation of the results as obtained is not shared by the Panel. Its assumptions were based on limited scientific data obtained from a single morphometric study (which was already found to be inadequate in the previous EFSA Opinion on bovine casings) and on limited and earlier data on the presence of PrPsc/infectivity in bovine gut after experimental oral BSE inoculation. There is uncertainty about the relative BSE risk of neural and lymphoid tissues in casings compared to CNS that might have significant impact on the calculated results of the DNV Report. The Panel notes that the DNV Report considers the individual human BSE exposure risk from bovine casings, excluding ileum, to be “very low”. However, when the upper confidence limits are taken into account, along with the uncertainties in key parameter assumptions, the calculated total human exposure per year in the EU from bovine casings, even when ileum is excluded (based on the calculated BSE prevalence in 2007) is 11.000 cattle oral ID50 units per year (when all casings would have been sourced in the UK) and about 1.000 cattle oral ID50 units per year (when all casings would have been sourced in the Netherlands) and therefore cannot be considered negligible. Thus the conclusion in the DNV report that sausage casings sourced from intestines of cattle in EU Member States would lead to a negligible risk for human consumption cannot be considered valid. Moreover, when considering other new relevant scientific information it is concluded that the previous EFSA assessment of the BSE related risk of bovine intestines after processing into natural sausage casings remains valid. The Panel recommends that future considerations on the risk in bovine casings should take into account the BSE prevalence in cattle at that time.

Published: 22 September 2009

http://www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1211902899454.htm?WT.mc_id=EFSAHL01&emt=1


SUMMARY

http://www.efsa.europa.eu/EFSA/Scientific_Opinion/biohaz_op_1317_bovine_intestines_summary_en,0.pdf?ssbinary=true


OPINION


snip...

6. Overview of current scientific knowledge on BSE risk in Bovine Intestines. The previous EFSA Opinion on BSE risk from bovine intestine summarised the scientific knowledge that was available until early 2007. Since then, additional publications have become available on a natural BSE case in Japan (Kimura and Haritani, 2008) and two experimental studies that examined

presence of PrPsc and/or infectivity in the intestines of cattle challenged orally with 100g (Espinosa et al., 2007; Hoffmann et al., 2007). Moreover, a new study performed by the VLA in the UK on PrPsc in BSE-infected cattle (Stack, 2009) and preliminary results from the German BSE pathogenesis study have recently be made available to EFSA and were also taken into account.

6.1. New experimental studies on intestines of BSE infected cattle

Espinosa et al. (2007) examined pooled tissues from 13 cattle inoculated at ages between 4 and 6 months and culled at ages between 24 and 39 months. Infectivity in Tgbov mice but not PrPsc by ELISA/WB was found in Peyer’s patches dissected from distal ileum at all ages. Hoffmann et al (2007) demonstrated PrPsc by IHC in Peyer’s patches of distal ileum in one of two preclinical animals sacrificed at 24 and 28 months post inoculation (mpi). Most recently, Arnold et al. (2009) estimated the titre of infectivity in the distal ileum from the incubation time found by bioassay in wild type mice. Over time, the infectivity in the distal ileum showed an initial increase up to 14-18 months post exposure, followed by a decrease, which was likely to be highly variable between animals. However, these estimates were based on mouse titration of brain material, while the incubation period to dose relationship may differ between brain and intestines (Robinson et al., 1990).

6.2. Infectivity of intestines in cattle with natural BSE infection

Data on presence of PrPsc or infectivity in intestines of natural BSE cases are sparse. The immunohistochemistry (IHC) and Western blot examinations of three BSE infected cattle detected in Japan in the course of active surveillance (but showing locomotor deficits) found PrPsc in distal ileum of two (by IHC confined to the myenteric plexus) (Iwata et al., 2006). No PrPsc was detected in Peyer’s patches of distal ileum, or in samples of other regions of small and large intestine, or in a range of other lymphoid tissues. Labelling of myenteric plexus was also detected in 9/29 confirmed field cases of BSE examined in the UK (Terry et al., 2003). Infectivity by wild-type mouse assay or the presence of PrPsc has not been found in the distal ileum, or other levels of intestine in a total of some six natural BSE cases studied (Fraser and Foster, 1994; Buschmann and Groschup 2005; Iwata et al., 2006). In one of these cases in Germany, however, infectivity was detected in the distal ileum by bioassay in TgBov XV mice (Buschmann and Groschup, 2005). More recently, another BSE case (94 months of age) in Japan showed definite or equivocal immunoreactivity in nerve cells of the myenteric plexus in ileum, caecum and colon, and in Schwann cells of the myenteric plexus in duodenum, jejunum, ileum, caecum and colon (Kimura and Haritani, 2008).

6.3. Study commissioned by ENSCA

This ENSCA commissioned study investigated the presence of BSE PrPsc in small intestines of cattle that had been orally challenged at 4-6 months of age with 100g or 1 g doses of BSE affected brain tissue. These animals were culled and examined 18-30 months post inoculation (p.i.). Three methods to identify PrPsc were applied: a commercial ELISA test, Western immunoblotting, and IHC. Results confirmed previous observations that PrPsc was mainly confined to lymphoid tissue of the ileum, whereas the duodenum was negative and no part of the enteric nervous system tested positive. The lymphoid tissue of the jejunum of one high-dosed animal tested positive. As expected, the low-dosed animals had a much lower frequency of positive ileum samples (1/18 vs. 15/18 in the high-dose group) and some longer incubation times (24 months in the one animal with positive ileum), whereas the high-dose group included animals positive at all ages examined.

As the ENSCA commissioned study was performed retrospectively on archival tissue, sampling was limited by availability, and the study authors themselves concede that “it is possible tissue sampling was not optimal” for duodenum and jejunum of low-dosed animals. The 1g-dosed group included 6 animals sampled at 18 months p.i., 6 at 24 months, and 6 at 30 months. The 100g-dosed group included 6 animals sampled for ileum at 18 months p.i., 6 at 24 months, and 6 at 30 months;

duodenum and jejunum, however, were sampled only in 2 animals each at 18, 24 and 30 months p.i., respectively. From each level of the intestine, three sections were examined by IHC per case. While at least two of the three sections of the ileum per case contained lymphoid follicles, in 36% of the duodenum cases, and in 39% of the jejunum cases lymphoid follicles were absent in any of the examined sections. The frequency of positive follicles per section ranged between 1% and 14% in ileum of the high-dose group, and 0,7% in the one positive ileum of the low-dose group, and was 6,7% and 11,1% in the two positive jejunum sections of one high-dosed animal.

Conclusions on the ENSCA commissioned study:

• This study confirms that detectable PrPsc is mainly confined to lymphoid tissue of the ileum in cattle orally challenged with 100g of BSE brain and culled at 18, 24 and 30 months postinoculation (p.i.)

• One out of 18 animals challenged orally with 1g of BSE brain was positive in ileum.

• One out of 18 animals challenged orally with 100g of BSE brain was positive in jejunum.

• The duodenum was always negative.

• However, the sampling in particular of duodenum and jejunum was limited and contained lymphoid tissue only in a part of sections examined.

• In contrast to previous reports on natural BSE cases in older animals, the enteric nervous system was always negative.

• In consideration of the previous EFSA opinion on bovine intestine that gives detailed advice for future studies, in particular concerning the lower frequency of lymphoid follicles in parts of the intestine other than the distal ileum, the present ENSCA commissioned study meets some but not all recommendations; in particular the mostly negative results obtained for jejunum and duodenum should not be over-interpreted when tissue sampling was limited.

6.4. New preliminary data on bovine intestine from the German BSE Pathogenesis study

In the German BSE pathogenesis study performed at the Friedrich-Loeffler-Institute (FLI), 56 Simmental cross-breed calves aged about four months were challenged orally with 100g brainstemhomogenate pooled out of clinically BSE diseased cattle. The infectivity load in the homogenate was about 106.1 ID50 (grams of tissue)-1 as determined by end-point titration in Tgbov XV mice (Buschmann & Groschup, 2005, Hoffmann et al., 2007). Furthermore, as controls, 18 calves were inoculated orally with a BSE-negative brainstem homogenate. Four to five animals were selected randomly and euthanised every four months. More than 150 tissue and body fluid samples were sampled at subsequent necropsies from each animal under TSE-sterile conditions.

After oral exposure with the TSE agent, previous studies had demonstrated consistently early prion accumulation in the gut associated lymphatic tissue, about six months post infection (mpi) in cattle (Terry et al., 2003), and at two mpi in scrapie infected sheep (van Keulen et al., 2002) and in 21 days old lambs (Andreoletti et al., 2002). In contrast to scrapie, the accumulation of PrPd in the distal ileum of BSE-infected cattle was confined to an only minor proportion of follicles respectively neurons/glial cells of the enteric nervous system (Terry et al., 2003).

Normally when performing IHC, a three micrometer section per paraffin block is used, reflecting a very small proportion of the tissue sample. Therefore a serial section procedure was newly established at the FLI to increase the total amounts of tissue structures examined per sample and consequently increasing the probability of detecting PrPsc accumulation. Thereby, five sections per paraffin block with a plane distance of about 25-30 µm were examined. Hence, a tissue depth of about 150-200 µm per block was screened for positive immunosignals. Additionally two different PrP-specific monoclonal antibodies, highly sensitive for the detection of bovine PrPsc were used.

According to this method, representative samples of the small intestine, in particular Peyer’s patches of the distal ileum but also the ileo-caecal junction from most of the infected animals of the German BSE Pathogenesis study were examined by IHC. From 4 mpi until 44 mpi in most animals (38/43), PrPsc was detectable, initially in the follicles of the Peyer’s patches and at later stages of the incubation period in the enteric nervous system too.

Conclusions on the German pathogenesis study

• With improved sampling, nearly all animals dosed with 100 g of BSE brain tissue showed PrPsc in distal ileum between 4 and 44 mpi, first in lymphoid tissue and later in enteric nervous system.

7. Review of the DNV report

7.1. Summary of the report

DNV makes an attempt to quantify the amount of BSE infectious load in bovine sausage casings. This is then extrapolated to the risk carried in an individual sausage, a normal persons risk per year and the overall exposure within the EU in a year. The key points of the DNV Report are as follows:

• The DNV Report assumes that the ileum is not used for the production of casings and is removed and discarded.

• The DNV Report is based on the assumption that potential infectivity in bovine intestine used for sausage casing production would be 2 logs less than in the ileum. Based on experimental data, the infectivity in the distal ileum was considered to be at a titre equivalent to that in the CNS at the late stage of infection. Thus infectivity in non-ileal parts of the intestines used for casings production was assumed to be 100 fold less than in the CNS.

• The DNV Report uses a value of 0.43g/m (obtained from Wijnker et al.) of casing to quantify the amount of lymphoid and neural tissue that might harbour infectivity in a sausage casing,

• The results of the DNV Report calculate that an exposure per person per year from bovine casings produced in the Netherlands “would be very low” even when a high consumption pattern like in Germany is assumed (upper range 7 x 10-6 cattle oral (CO) ID50 units). For casings sourced in the UK, the exposure would be about one log higher.

• When the calculated total amount of cumulative human exposure per year in the EU is considered, the following scenario emerges: 11.000 CO ID50 units per year when all casings would have been sourced in the UK, and about 1.000 CO ID50 units when all casings would have been sourced in the Netherlands, a country with an about average prevalence of BSE in the EU4.

4 How can the output of the DNV calculations be interpreted in terms of potential human infections? If we follow, as in the previously adopted EFSA Opinions on Tallow and MBM (EFSA 2005 a and b) the cautionary advice of the original QRA WG and assume the species barrier is 1 as a worst case scenario, then there would be up to 5500 infected person in the EU per year in the first scenario, and up to about 500 in the second. This would have to assume a linear dose-response curve of infectivity at very low doses. If the species barrier was given a more realistic value obtained from the analysis carried out on the exposure of the British population to the BSE agent (EFSA, 2006) of around 1000 - 4000, this would mean that there might be up to around 1 to 5 infected person in the EU per year in the first scenario, and less than 1 in the second.

snip... see full text ;


http://www.efsa.europa.eu/cs/BlobServer/Scientific_Opinion/biohaz_op_1317_bovine_intestines_en,0.pdf?ssbinary=true




----- Original Message -----
From: Terry S. Singeltary Sr.
To: fdadockets@oc.fda.gov
Sent: Wednesday, September 07, 2005 9:44 PM
Subject: Use of Materials Derived From Cattle in Human Food and Cosmetics [Docket No. 2004N-0081] RIN 0910-AF47

Greetings FDA,


I would kindly like to comment on ; Use of Materials Derived From Cattle in Human Food and Cosmetics [Docket No. 2004N-0081] RIN 0910-AF47 SUMMARY: The Food and Drug Administration (FDA) is amending the interim final rule on use of materials derived from cattle in human food and cosmetics published in the Federal Register of July 14, 2004. In the July 14, 2004, interim final rule, FDA designated certain materials from cattle, including the entire small intestine, as ``prohibited cattle materials'' and banned the use of such materials in human food, including dietary supplements, and in cosmetics. FDA is taking this action in response to comments received on the interim final rule. Information was provided in comments that persuaded the agency that the distal ileum, one of three portions of the small intestine, could be consistently and effectively removed from the small intestine, such that the remainder of the small intestine, formerly a prohibited cattle material, could be used for human food or cosmetics. We (FDA) are also clarifying that milk and milk products, hide and hide-derived products, and tallow derivatives are not prohibited cattle materials. Comments also led the agency to reconsider the method cited in the interim final rule for determining insoluble impurities in tallow and to cite instead a method that is less costly to use and requires less specialized equipment. FDA issued the interim final rule to minimize human exposure to materials that scientific studies have demonstrated are highly likely to contain the bovine spongiform encephalopathy (BSE) agent in cattle infected with the disease. FDA believes that the amended provisions of the interim final rule provide the same level of protection from human exposure to the agent that causes BSE as the original provisions. ...

I would kindly like to submit the following ;

I find it very very disturbing that FDA now takes the position;

>>>Information was provided in comments that persuaded the agency that the distal ileum, one of three portions of the small intestine, could be consistently and effectively removed from the small intestine, such that the remainder of the small intestine, formerly a prohibited cattle material, could be used for human food or cosmetics. <<< href="http://stanford.wellsphere.com/cjd-article/use-of-materials-derived-from-cattle-in-human-food-and-cosmetics-docket-no-2004n-0081-rin-0910-af47/641209">http://stanford.wellsphere.com/cjd-article/use-of-materials-derived-from-cattle-in-human-food-and-cosmetics-docket-no-2004n-0081-rin-0910-af47/641209




http://stanford.wellsphere.com/cjd-article/docket-no-03-080-1-usda-issues-proposed-rule-to-allow-live-animal-imports-from-canada/641254




http://cjdmadcowbaseoct2007.blogspot.com/2008/04/use-of-materials-derived-from-cattle-in.html





THE SEVEN SCIENTIST REPORT ***



The tissue distribution of infectivity in BSE infected cattle has primarily been determinedby 3 studies conducted in the United Kingdom all of which had limitations.In two of the studies, bioassays were done in mice which are at least 1000 fold lesssensitive to BSE infection than cattle themselves. Only higher titers of infectivity can bedetected by this method. These investigations found infectivity in the brain, spinal cord,retina, trigeminal ganglia, dorsal root ganglia, distal ileum and bone marrow (the bonemarrow finding was from one animal). Infectivity was found in distal ileum ofexperimentally infected calves beginning six months after challenge and continuing atother intervals throughout life. (Wells et. al., 1994; 1998). The bioassay study in calveshas produced similar results and in addition infectivity has been found in tonsil. Thestudy is still in progress. Another project has found infectivity in the lymphoid tissue ofthird eyelid from naturally infected animals. (Dr. Danny Matthews, UK DEFRA,personal communication).While bioassay in cattle is far preferable to mice in terms of sensitivity, cattlenevertheless present their own limitations in terms of the long incubation time and thelimited number of animals that can be used for assay compared to rodents. As aconsequence the significance of the negative finding for many tissues is questionable. Infact, by the end of 2004 there was increasing evidence in species other than cattle thatperipheral nerves and muscle have infectivity. (Bosque et al., 2002; Glatzel et al.,2003;Bartz et al., 2002; Androletti et al., 2004; Mulcahy et al., 2004; Thomzig et al.,2003; Thomzig et al., 2004)In some of these species, studies indicate that the agent migrates to the brain and spinalcord, replicates to high levels in the CNS and then spreads centrifugally from the spinalcord back down through the spinal neurons to the junction of the nerves and muscle intothe muscle cells themselves. A recent German study (Buschmann and Groschup, 2005)examined nerves and muscle from a cow naturally infected with BSE and found thatinfectivity was present in several peripheral nerves and one muscle. The method ofdetection was bioassay in bovinized transgenic mice that show the same or greatersensitivity to transmission of BSE as cattle. This research concurs with findings byJapanese scientists that BSE infectivity is present in peripheral nerves at least in theclinical stage of disease.It is our opinion that there is increasing evidence that the pathogenesis of BSE might notbe entirely different from TSEs in other species at the point of clinical disease in thatthere is peripheral involvement. We feel that the studies as reported above have merit.The current studies not only re-enforce the risk of down and deadstock but also appear toprovide additional information that these animals may be a potential source of greaterlevels of infectivity into the feed system.In the event that FDA may confer with USDA about the risks associated with peripheralnerves we want to point out one issue. In the recent publication of the final rule on theDeleted: SRMs, as defined by theUSDA, are tissues which, in a BSEinfected animal, are known to eitherharbor BSE infectivity or to be closelyDeleted: Rendered with other slaughterInserted: pose a risk of BSEDeleted: may introduce BSE infectivityDeleted: aDeleted: source ofDeleted: animal feedDeleted: perfectly balanced animalInserted: perfectly balanced animalDeleted: contaminationDeleted: they were a major source ofDeleted: .Deleted: SRMs include tissues that areDeleted: does not totally eliminate it.Inserted: doesDeleted: This is significantDeleted: This residual infectivity stillInserted: SRMs include tissues that areInserted: , which are bones, areDeleted: which encaseDeleted: the brain and spinal cordDeleted:Inserted: tissue that they encase andDeleted: , respectively, can be assumedDeleted: RenderingDeleted: Conventional renderingDeleted: willDeleted: reduces infectivity fromDeleted: willInserted: Rendered with otherInserted: they were a major source ofInserted: Conventional renderingInserted: sInserted: from contaminated tissuesInserted: This residual infectivity stillDeleted: (four to six years for high titerDeleted:Deleted: alreadyDeleted: Kovacs et al., 2004;Deleted: pDeleted: 14... [25]... [19]... [26]... [24]... [29]... [31]... [33]... [23]... [30]... [32]... [21]... [34]... [22]... [35]... [17]... [27]... [18]... [20]... [28]FDA Proposed Rule December 20, 2005importation of whole cuts of boneless beef from Japan, 9 CFR Part 94 [Docket No. 05-004-2] RIN 0579-AB93, we disagree with the interpretation provided by USDA, APHIS.APHIS seems to discount the studies conducted by Groschup et al. 2005. on the basis thatthe transgenic mouse bioassay that they used may be too sensitive. In taking this positionthey have failed to realize that the point of an assay is to reveal in which tissues theinfectivity resides and its relative concentration to brain or spinal cord. For this purpose,no assay can be too sensitive. Of course, the probability of an actual infection will beaffected by the efficiency of infection which will be a function of dose, route of exposureand any host barrier effects that are present.We would also like to point out a factual error in the conclusion. APHIS states, “Giventhese factors, APHIS has determined that the finding of BSE infectivity in facial and sciatic nervesof the transgenic mice is not directly applicable to cattle naturally infected with BSE. Therefore,we do not consider it necessary to make any adjustments to the risk analysis for this rulemakingor to extend the comment period to solicit additional public comment on this issue.” It is incorrectthat the infectivity was found in the peripheral nerves of transgenic mice. The peripheralnerves were harvested from a cow naturally infected with BSE. Transgenic mice wereused as a bioassay model.From [Docket No. 05-004-2] RIN 0579-AB93:“Peripheral NervesIssue: Two commenters stated that the underlying assumption of the proposed rule, that wholecuts of boneless beef from Japan will not contain tissues that may carry the BSE agent, is nolonger valid because researchers have found peripheral nervous system tissues, including facialand sciatic nerves, that contain BSE infectivity.\2\ One of these commenters requested APHIS toexplain whether and what additional mitigation measures are needed to reduce the risks thatthese tissues may be present in Japanese beef. This commenter further requested an additionalcomment period to obtain public comments to treat this new scientific finding.



http://www.fda.gov/ohrms/dockets/dockets/02n0273/02n-0273-EC244-Attach-1.pdf


================================


Greetings again APHIS ET AL,

THIS is not correct. IN fact, there are several factors i would like to kindly address.

Muscle tissue has recently been detected with PrPSc in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve) of the 11th BSE cow in Japan (Yoshifumi Iwamaru et al). also recently, Aguzzi et al Letter to the Editor Vet Pathol 42:107-108 (2005), Prusiner et al CDI test is another example of detection of the TSE agent in muscle in sCJD, Herbert Budka et al CJD and inclusion body myositis: Abundant Disease-Associated Prion Protein in Muscle, and older studies from Watson Meldrum et al Scrapie agent in muscle - Pattison I A (1990), references as follow ;

PrPSc distribution of a natural case of bovine spongiform encephalopathy

Yoshifumi Iwamaru, Yuka Okubo, Tamako Ikeda, Hiroko Hayashi, Mori- kazu Imamura, Takashi Yokoyama and Morikazu Shinagawa Priori Disease Research Center, National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba 305-0856 Japan mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000423/!x-usc:mailto:gan@affrc.go.jp

Abstract

Bovine spongiform encephalopathy (BSE) is a disease of cattle that causes progressive neurodegeneration of the central nervous system. Infectivity of BSE agent is accompanied with an abnormal isoform of prion protein (PrPSc).

The specified risk materials (SRM) are tissues potentially carrying BSE infectivity. The following tissues are designated as SRM in Japan: the skull including the brain and eyes but excluding the glossa and the masse- ter muscle, the vertebral column excluding the vertebrae of the tail, spinal cord, distal illeum. For a risk management step, the use of SRM in both animal feed or human food has been prohibited. However, detailed PrPSc distribution remains obscure in BSE cattle and it has caused con- troversies about definitions of SRM. Therefore we have examined PrPSc distribution in a BSE cattle by Western blotting to reassess definitions of SRM.

The 11th BSE case in Japan was detected in fallen stock surveillance. The carcass was stocked in the refrigerator. For the detection of PrPSc, 200 mg of tissue samples were homogenized. Following collagenase treatment, samples were digested with proteinase K. After digestion, PrPSc was precipitated by sodium phosphotungstate (PTA). The pellets were subjected to Western blotting using the standard procedure. Anti-prion protein monoclonal antibody (mAb) T2 conjugated horseradish peroxidase was used for the detection of PrPSc.

PrPSc was detected in brain, spinal cord, dorsal root ganglia, trigeminal ganglia, sublingual ganglion, retina. In addition, PrPSc was also detected in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve). Our results suggest that the currently accepted definitions of SRM in BSE cattle may need to be reexamined. ...

179 T. Kitamoto (Ed.) PRIONS Food and Drug Safety

================

ALSO from the International Symposium of Prion Diseases held in Sendai, October 31, to November 2, 2004;

Bovine spongiform encephalopathy (BSE) in Japan

snip...

"Furthermore, current studies into transmission of cases of BSE that are atypical or that develop in young cattle are expected to amplify the BSE prion"

NO. Date conf. Farm Birth place and Date Age at diagnosis

8. 2003.10.6. Fukushima Tochigi 2001.10.13. 23

9. 2003.11.4. Hiroshima Hyogo 2002.1.13. 21

Test results

# 8b, 9c cows Elisa Positive, WB Positive, IHC negative, histopathology negative

b = atypical BSE case

c = case of BSE in a young animal

b,c, No PrPSc on IHC, and no spongiform change on histology

International Symposium of Prion Diseases held in Sendai, October 31, to November 2, 2004.

The hardback book title is 'PRIONS' Food and Drug Safety

T. Kitamoto (Ed.) Tetsuyuki Kitamoto Professor and Chairman Department of Prion Research Tohoku University School of Medicine 2-1 SeiryoAoba-ku, Sendai 980-8575, JAPAN TEL +81-22-717-8147 FAX +81-22-717-8148 e-mail; mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000423/!x-usc:mailto:kitamoto@mail.tains.tohoku.ac.jp Symposium Secretariat Kyomi Sasaki TEL +81-22-717-8233 FAX +81-22-717-7656 e-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000423/!x-usc:mailto:kvomi-sasaki@mail.tains.tohoku.ac.ip

================================




snip...end...full text ;


http://www.regulations.gov/fdmspublic/ContentViewer?objectId=0900006480086ebc&disposition=attachment&contentType=msw6



Response to Public Commentson theHarvard Risk Assessment of Bovine SpongiformEncephalopathy Update,October 31, 2005

snip...

RESPONSE TO COMMENTS FROM TERRY S. SINGELTARY SR.

Comment #1: SINCE the first Harvard BSE Risk Assessment was so flawed and fraught with error after the PEER REVIEW assessment assessed this fact, how do you plan on stopping this from happening again, will there be another peer review with top TSE Scientists, an impartial jury so-to-speak, to assess this new and updated Harvard BSE/TSE risk assessment and will this assessment include the Atypical TSE and SRM issues?

Response: The original (October 2003) and the revised (October 2005) Harvard BSE risk assessments underwent external peer review. Subsequently, revisions were made to the analysis. In the most recent review, the most significant revisions have been:

1) theaddition of explicit modeling of the poultry litter pathway for the potential recycling ofbovine protein into cattle feed; and

2) a decrease in the assumed effectiveness of antemortem inspection in the identification of animals with BSE.

Comment #2: WITH A RECENT NATION WIDE MAD COW FEED BAN RECALL in the past few months that consisted of some 10,878.06 TONS, then another Mad Cow feed ban warning letter in May, IT should seem prudent to ask why our feed bans continue to fail in 2006, and continue to fail today?

Response: This question about feed bans is a matter for policy. As such, it is not addressed in this response.

Comment #3: WHY still now only partial ruminant feed ban, with the fact that now we seem to have 3 cases of nvCJD to humans i.e. human bovine TSE that were responsible from blood, and the fact the last 2 mad cows documented in the USA were that of an Atypical strain, would it not seem prudent to remove blood as well from ruminant feed?WOULD it not seem prudent to improve and expand the SRM list now? as per your ownthinking; If transmission occurs, tissue distribution comparisons will be made between cattle infected with the atypical BSE isolate and the U.S. BSE isolate. Differences in tissue distribution could require new regulations regarding specific risk material (SRM) removal.

Response: This comment pertains to policy. As such, it is not addressed here.

Comment #4: WHAT does USDA/FDA ET AL intend to do about the risks of atypical BSE/TSE in cattle now that infectivity shows in tissue samples other than CNS in Japan, the fact now that the last Texas mad cow and that last mad cow in Alabama were indeed of the atypical strain, the fact that the studies long ago in Mission, Texas of USA sheep scrapie transmission to the USA bovine, which proved an 'atypical tse' in the USA bovine, the fact also that USDA/FDA are still floundering on the last SRM regulations, but with the BASE strain now in cattle that is not similar to nvCJD, but very similar to the sporadic CJD, and sporadic CJD has tripled in the last few years in the USA. WHAT do you plan to do to protect human health from these atypical strains of TSE, in relations to SRMs ?

- 19 -

--------------------------------------------------------------------------------

Page 20

Response: The BSE risk assessment simulation model characterizes the disease history of BSE, including the agent’s spread within the body of the animal over time. It also quantifies the agent’s persistence during the feed manufacture process, and ultimately the agent’s ability to cause disease in other exposed animals. There is no definitive evidence that these properties differ substantially for atypical BSE strains, compared to the typical BSE agent.

Comment #5: THE 2004 Enhanced BSE surveillance program, that tested all those cows, but then we found just how terribly flawed the program was, from testing protocols, to testing the most likely to have BSE i.e. high risk, to the geographical distribution of the testing and high risk areas, to letting the tissue samples of one mad cow sit on a shelf for 7+ months and then having to have an act of Congress to ever get that cow finally confirmed, to that other Texas mad cow they decided to not even bother testing at all, just rendered that very suspect cow, too suspect to test evidently, back to that Alabama madcow that they could only give a guess as to age with dentition where we all know that the age of that cow was so close to 10 years it could have been 9 years 7 months to 10 years 3 months, thus possibly being an BAPB i.e. USA 'born after partial ban', to all those rabies suspect cows that did not have rabies, and DID NOT get tested for BSE/TSE in that June 2004 enhanced surveillance program, even though the common lay person knows the suspect rabies negative cows are suppose to be BSE/TSE tested, how does one correct all these blatant failures and will they be corrected?

Response: This comment pertains to policy. As such, it is not addressed here.

Comment #6: WHAT happened to the test results and MOUSE BIO-ASSAYS of those imported sheep from Belgium that were confiscated and slaughtered from the Faillace's, what sort of TSE did these animals have?

Response: It is not clear how the test results referred to in this comment are relevant to the Harvard BSE Risk Assessment Update. Sheep were not considered in the risk assessment.

Comment #7: WHY is it that the Farm of the Mad Sheep of Mad River Valley were quarantined for 5 years, but none of these farms from Texas and Alabama with Atypical TSE in the Bovine, they have not been quarantined for 5 years, why not, with the real risk of BSE to sheep, whom is to say this was not BSE ?

Response: This comment pertains to policy. As such, it is not addressed here.

- 20 -

snip...END

FULL TEXT ;


http://www.fsis.usda.gov/PDF/BSE_Risk_Assess_Response_Public_Comments.pdf



October 31, 2002
Review of the Evaluation of the Potential for Bovine Spongiform Encephalopathy in the United States
Conducted by the Harvard Center for Risk Analysis, Harvard School of Public Health and Center for Computational Epidemiology, College of Veterinary Medicine, Tuskegee University


http://www.fsis.usda.gov/oa/topics/BSE_Peer_Review.pdf



September 13,2004 USDA, FSTS Docket Clerk 300 12* Street, SW Room 102, Cotton Annex Washington, DC 20250 04-021ANPR 04-021ANPR-70 Richard L. Crawford

Re: Docket No: 04-02 1 ANPR Federal Measures to Mitigate BSE Risks: Considerations for Further Action

Dear Sir or Madame:

On behalf of McDonald’s Corporation, which operates more than 13,000 restaurants in the United States, we appreciate the opportunity to submit comments to this very important Advance Notice of Proposed Rulemaking (ANPRM). 69 Fed. Reg. 42288 (July 14,2004).

In previous comments submitted to FSIS regarding the removal of SRI&, McDonalds fully supported this rule and its immediate implementation. The removal of SRMs from human food is the primary firewall to protect the US consumer from being exposed to the BSE agent. While we applaud the requirement for SRM removal, we feel that it is equally important for FSIS to insure that each slaughterplant which processes cattle have systems in place which prevent cross contamination between edible tissue and SRMs. This should include but not be limited to the use of separate equipment, such as knives, blades, etc. where appropriate. In addition, it is also important that appropriate and effective disinfection procedures for equipment used to handle SRMs be developed and approved for use.

It is our opinion that requiring SRM removal without a procedure to prevent cross contamination is inadequate as a protective public health measure. The TSE agents @ions) are sticky and highly resistant to disinfection. If SRMs such as brain and spinal cord are allowed to contact equipment and other surfaces such as deboning tables which then are used to handle and process edible tissue this could allow contamination and negates the intention of the ban. This is true not only in plants slaughtering fed cattle both under and over 30 months but also in plants slaughtering predominately older cattle. It is important that measure be taken to prevent cross contamination between carcasses and SRms in the cull plants. McDonalds requires their suppliers to prevent cross contamination and audits against certain measurable standards such as requiring spinal cord to bc removed on the kill floor. We would be willing to share these standards with FSIS as an example.

FSIS Docket No. 04-02 1 ANPR

McDonalds again recommends that dura (the covering around the brain and spinal cord) be added to the list of SRMs. While skull and vertebral column are included as SRMs, dura is not. If dura is not removed prior to processing on the fabrication floor, it may come loose and be incorporated into ground product. Bovine dura was never tested for infectivity. It was assumed that due to direct contact with spinal cord, it may serve as a vehicle to transmit disease. In addition, human dura has been the source of human to human transmission of Creutzfeldt-Jakob Disease (CJD). (personal communication - Dr. Danny Matthews, UK, VLA) Our ISAC committee recommended that McDonalds add the removal of dura as a specification in the production of our product.

McDonalds urges the USDA to make the appropriate adjustments in the SRM ban if new scientific findings and/or the results of the increased surveillance warrant a change. In regards to imported meat products from other countries, McDonalds suggests that no SRM exemption be made for countries based on BSE risk. The long incubation period and limited surveillance in many countries can limit the ability to accurately determine risk. Also, the risk level of a country could potentially change over night if the trading patterns of a country changed. It seems logistically impossible to maintain a system which could continually monitor the world’s trading patterns. In addition, science has not provided all of the answers in regards to the transmission of BSE. Requiring SRMs to be removed from imported products for human food is prudent. If the US would wait until disease is confirmed the exposure would already have occurred.

Thank you for the opportunity to comment on these very important issues.

Richard L. Crawford Corporat,e Vice President, Government Relations McDonalds Corporation 1 Kroc Drive Oak Brook, Illinois 60523 FSIS Docket No. 04-021ANPR



http://www.fda.gov/ohrms/dockets/dailys/04/sep04/092104/04n-0264-c00140-vol22.pdf



http://www.fsis.usda.gov/OPPDE/Comments/03-038IF/03-038IF-15.pdf



Dockets Entered on December 22, 20052005D-0330, Guidance for Industry and FDA Review Staff on Collection of Plateletsby Automated ... EC 203, McDonald's Restaurants Corporation, Vol #:, 34 ...


http://www.fda.gov/ohrms/dockets/dailys/05/Dec05/122205/122205.htm


03-025IF 03-025IF-631 Linda A. Detwiler [PDF]Page 1. 03-025IF 03-025IF-631 Linda A. Detwiler Page 2. Page 3. Page 4.Page 5. Page 6. Page 7. Page 8. Page 9. Page 10. Page 11. Page 12.

www.fsis.usda.gov/OPPDE/Comments/03-025IF/03-025IF-631.pdf


- Text Version
03-025IF 03-025IF-634 Linda A. Detwiler [PDF]Page 1. 03-025IF 03-025IF-634 Linda A. Detwiler Page 2.Page 3. Page 4. Page 5. Page 6. Page 7. Page 8.

www.fsis.usda.gov/OPPDE/Comments/03-025IF/03-025IF-634.pdf


- Text Version[ More results from www.fsis.usda.gov/OPPDE/Comments/03-025IF/ ]
Page 1 of 17 9/13/2005 [PDF]... 2005 6:17 PM To: fsis.regulationscomments@fsis.usda.gov Subject: [Docket No. 03-025IFA]FSIS Prohibition of the Use of Specified Risk Materials for Human Food ...

www.fsis.usda.gov/OPPDE/Comments/03-025IFA/03-025IFA-2.pdf


- Text Version 03-025IFA 03-025IFA-6 Jason Frost [PDF]... Zealand Embassy COMMENTS ON FEDERAL REGISTER 9 CFR Parts 309 et al [Docket No. 03-025IF] Prohibition of the Use of Specified Risk Materials for Human Food and ...

www.fsis.usda.gov/OPPDE/Comments/03-025IFA/03-025IFA-6.pdf


- Text Version


http://www.fsis.usda.gov/Search/Search_Results/Index.asp?q=03-025IF&mode=simple&num=10&as_occt=any&restrict=FSIS_DOCKET_COMMENTS


In its opinion of 7-8 December 2000 (EC 2000), the SSC ... [PDF]Page 1. Linda A. Detwiler, DVM 225 Hwy 35 Red Bank, New Jersey 07701 Phone: 732-741-2290Cell: 732-580-9391 Fax: 732-741-7751 June 22, 2005 FSIS Docket Clerk US ...


www.fsis.usda.gov/OPPDE/Comments/03-025IF/03-025IF-589.pdf


http://www.fsis.usda.gov/OPPDE/Comments/03-025IF/03-025IF-589.pdf



Page 1 of 17 9/13/2005 [PDF]
... Page 1 of 17 From: Terry S. Singeltary Sr. [flounder9@verizon.net] Sent: Thursday,September 08, 2005 6:17 PM To: fsis.regulationscomments@fsis.usda.gov Subject ...



www.fsis.usda.gov/OPPDE/Comments/03-025IFA/03-025IFA-2.pdf


http://www.fsis.usda.gov/OPPDE/Comments/03-025IFA/03-025IFA-2.pdf



03-025IF 03-025IF-618 Richard L. Crawford [PDF]Page 1. 03-025IF 03-025IF-618 Richard L. CrawfordPage 2. Page 3. Page 4.


www.fsis.usda.gov/OPPDE/Comments/03-025IF/03-025IF-618.pdf


http://www.fsis.usda.gov/OPPDE/Comments/03-025IF/03-025IF-618.pdf


03-038IF 03-038IF-15 Richard L. Crawford [PDF]Page 1. 03-038IF 03-038IF-15 Richard L. CrawfordPage 2. Page 3. Page 4.

www.fsis.usda.gov/OPPDE/Comments/03-038IF/03-038IF-15.pdf


http://www.fsis.usda.gov/OPPDE/Comments/03-038IF/03-038IF-15.pdf


http://www.fsis.usda.gov/OPPDE/Comments/03-025IF/03-025IF-634.pdf



2009 UPDATE ON ALABAMA AND TEXAS MAD COWS


http://bse-atypical.blogspot.com/2006/08/bse-atypical-texas-and-alabama-update.html



TSS

PrPTSE in muscle-associated lymphatic tissue during the preclinical stage of mice orally-infected with BSE

Published online ahead of print on 17 June 2009 as doi:10.1099/vir.0.010801-0 J Gen Virol (2009), DOI 10.1099/vir.0.010801-0 © 2009 Society for General Microbiology

PrPTSE in muscle-associated lymphatic tissue during the preclinical stage of mice orally-infected with BSE

Franco Cardone1,6, Achim Thomzig2, Walter J Schulz-Schaeffer3, Angelina Valanzano1, Marco Sbriccoli1, Hanin Abdel-Haq1, Silvia Graziano1, Maria Puopolo1, Paul Brown4, Michael Beekes5 and Maurizio Pocchiari1

1 Istituto Superiore di Sanità; 2 Robert Koch-Institut, Berlin, Germany; 3 Georg-August University, Goettingen, Germany; 4 None; 5 RKI

6 E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:franco.cardone@iss.it

The involvement of muscles in the pathogenesis of transmissible spongiform encephalopathies (TSE) is irregular and unpredictable. We show that the TSE-specific protein (PrPTSE) is present in muscles of mice fed with a mouse-adapted strain of bovine spongiform encephalopathy (BSE) as early as 100 days post infection, corresponding to about one third of the incubation period. The proportion of mice with positive muscles and the number of muscles involved increased as infection progressed, but never attained more than limited distribution even at the clinical stage of disease. The appearance of PrPTSE in muscles during the pre-clinical stage of disease was likely due to the haematogenous/lymphatic spread of infectivity from the gastro-intestinal tract to lymphatic tissues associated with muscles, whereas in symptomatic animals the presence of PrPTSE in the nervous system, in neuromuscular junctions, and in muscle fibers suggests a centrifugal spread from the CNS as already observed in other TSE models.

Received 4 February 2009; accepted 17 June 2009.



http://vir.sgmjournals.org/cgi/content/abstract/vir.0.010801-0v1




Greetings,



some past studies ;




Muscle tissue has recently been detected with PrPSc

in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve) of the 11th BSE

cow in Japan (Yoshifumi Iwamaru et al). also recently, Aguzzi et al Letter to the Editor

Vet Pathol 42:107-108 (2005), Prusiner et al CDI test is another example of detection

of the TSE agent in muscle in sCJD, Herbert Budka et al CJD and inclusion body myositis:

Abundant Disease-Associated Prion Protein in Muscle, and older studies from Watson

Meldrum et al Scrapie agent in muscle - Pattison I A (1990), references as follow ;

PrPSc distribution of a natural case of bovine

spongiform encephalopathy

Yoshifumi Iwamaru, Yuka Okubo, Tamako Ikeda, Hiroko Hayashi, Mori-

kazu Imamura, Takashi Yokoyama and Morikazu Shinagawa

Priori Disease Research Center, National Institute of Animal Health, 3-1-5

Kannondai, Tsukuba 305-0856 Japan mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:gan@affrc.go.jp


Abstract


Bovine spongiform encephalopathy (BSE) is a disease of cattle that causes

progressive neurodegeneration of the central nervous system. Infectivity

of BSE agent is accompanied with an abnormal isoform of prion protein

(PrPSc).

The specified risk materials (SRM) are tissues potentially carrying BSE

infectivity. The following tissues are designated as SRM in Japan: the

skull including the brain and eyes but excluding the glossa and the masse-

ter muscle, the vertebral column excluding the vertebrae of the tail, spinal

cord, distal illeum. For a risk management step, the use of SRM in both

animal feed or human food has been prohibited. However, detailed

PrPSc distribution remains obscure in BSE cattle and it has caused con-

troversies about definitions of SRM. Therefore we have examined PrPSc

distribution in a BSE cattle by Western blotting to reassess definitions of

SRM.

The 11th BSE case in Japan was detected in fallen stock surveillance.

The carcass was stocked in the refrigerator. For the detection of PrPSc,

200 mg of tissue samples were homogenized. Following collagenase

treatment, samples were digested with proteinase K. After digestion,

PrPSc was precipitated by sodium phosphotungstate (PTA). The pellets

were subjected to Western blotting using the standard procedure.

Anti-prion protein monoclonal antibody (mAb) T2 conjugated horseradish

peroxidase was used for the detection of PrPSc.

PrPSc was detected in brain, spinal cord, dorsal root ganglia, trigeminal

ganglia, sublingual ganglion, retina. In addition, PrPSc was also detected

in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve).

Our results suggest that the currently accepted definitions of SRM in

BSE cattle may need to be reexamined. ...

179

T. Kitamoto (Ed.)

PRIONS

Food and Drug Safety


================


ALSO from the International Symposium of Prion Diseases held in Sendai, October 31, to

November 2, 2004;

Bovine spongiform encephalopathy (BSE) in Japan

snip...

"Furthermore, current studies into transmission of cases of BSE that are

atypical or that develop in young cattle are expected to amplify the BSE

prion"

NO. Date conf. Farm Birth place and Date Age at diagnosis

8. 2003.10.6. Fukushima Tochigi 2001.10.13. 23

9. 2003.11.4. Hiroshima Hyogo 2002.1.13. 21

Test results

# 8b, 9c cows Elisa Positive, WB Positive, IHC negative, histopathology

negative

b = atypical BSE case

c = case of BSE in a young animal

b,c, No PrPSc on IHC, and no spongiform change on histology

International Symposium of Prion Diseases held in Sendai, October 31, to

November 2, 2004.

The hardback book title is 'PRIONS' Food and Drug Safety

T. Kitamoto (Ed.)

Tetsuyuki Kitamoto

Professor and Chairman

Department of Prion Research

Tohoku University School of Medicine

2-1 SeiryoAoba-ku, Sendai 980-8575, JAPAN

TEL +81-22-717-8147 FAX +81-22-717-8148

e-mail; mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:kitamoto@mail.tains.tohoku.ac.jp

Symposium Secretariat

Kyomi Sasaki

TEL +81-22-717-8233 FAX +81-22-717-7656

e-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:kvomi-sasaki@mail.tains.tohoku.ac.ip


================================




http://www.regulations.gov/fdmspublic/ContentViewer?objectId=0900006480086ebc&disposition=attachment&contentType=msw6




http://www.scribd.com/doc/1490709/USDA-200600111



Detection and Localization of PrPSc in the Skeletal Muscle


Thu Mar 2, 2006 10:40 70.110.86.250

© 2006 American Society for Investigative Pathology

Detection and Localization of PrPSc in the Skeletal Muscle of Patients with Variant, Iatrogenic, and Sporadic Forms of Creutzfeldt-Jakob Disease Alexander H. Peden, Diane L. Ritchie, Mark W. Head and James W. Ironside From the National Creutzfeldt-Jakob Disease Surveillance Unit and Division of Pathology, School of Molecular and Clinical Medicine, University of Edinburgh, Western General Hospital, Edinburgh, United Kingdom

Variant Creutzfeldt-Jakob disease (vCJD) differs from other human prion diseases in that the pathogenic prion protein PrPSc can be detected to a greater extent at extraneuronal sites throughout the body, principally within lymphoid tissues. However, a recent study using a high-sensitivity Western blotting technique revealed low levels of PrPSc in skeletal muscle from a quarter of Swiss patients with sporadic CJD (sCJD). This posed the question of whether PrPSc in muscle could also be detected in vCJD, sCJD, and iatrogenic (iCJD) patients from other populations. Therefore, we have used the same high-sensitivity Western blotting technique, in combination with paraffin-embedded tissue blotting, to screen for PrPSc in muscle tissue specimens taken at autopsy from 49 CJD patients in the United Kingdom. These techniques identified muscle PrPSc in 8 of 17 vCJD, 7 of 26 sCJD, and 2 of 5 iCJD patients. Paraffin-embedded tissue blotting analysis showed PrPSc in skeletal muscle in localized anatomical structures that had the morphological and immunohistochemical characteristics of nerve fibers. The detection of PrPSc in muscle tissue from all forms of CJD indicates the possible presence of infectivity in these tissues, suggesting important implications for assessing the potential risk of iatrogenic spread via contaminated surgical instruments.



http://ajp.amjpathol.org/cgi/content/abstract/168/3/927



EMBO Rep. 2003 May; 4(5): 530–533. Published online 2003 April 11. doi: 10.1038/sj.embor.embor827. PMCID: PMC1319182

Copyright © 2003, European Molecular Biology Organisation Scientific Report Widespread PrPSc accumulation in muscles of hamsters orally infected with scrapie Achim Thomzig,1 Christine Kratzel,1 Gudrun Lenz,1 Dominique Krüger,1 and Michael Beekes1a 1Robert Koch-Institut, P26, Nordufer 20, D-13353 Berlin, Germany aTel: +49 30 4547 2396; Fax: +49 30 4547 2609; Email: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:BeekesM@rki.de Received February 13, 2003; Revised March 11, 2003; Accepted March 13, 2003. This article has been cited by other articles in PMC.

AbstractScrapie, bovine spongiform encephalopathy and chronic wasting disease are orally communicable, transmissible spongiform encephalopathies (TSEs). As zoonotic transmissions of TSE agents may pose a risk to human health, the identification of reservoirs for infectivity in animal tissues and their exclusion from human consumption has become a matter of great importance for consumer protection. In this study, a variety of muscles from hamsters that were orally challenged with scrapie was screened for the presence of a molecular marker for TSE infection, PrPSc (the pathological isoform of the prion protein PrP). Sensitive western blotting revealed consistent PrPSc accumulation in skeletal muscles from forelimb and hindlimb, head, back and shoulder, and in tongue. Previously, our animal model has provided substantial baseline information about the peripheral routing of infection in naturally occurring and orally acquired ruminant TSEs. Therefore, the findings described here highlight further the necessity to investigate thoroughly whether muscles of TSE-infected sheep, cattle, elk and deer contain infectious agent



http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1319182



Prions in Skeletal Muscles of Deer with Chronic Wasting Disease


Rachel C. Angers,1* Shawn R. Browning,1*† Tanya S. Seward,2 Christina J. Sigurdson,4‡ Michael W. Miller,5 Edward A. Hoover,4 Glenn C. Telling1,2,3§

1Department of Microbiology, Immunology and Molecular Genetics, 2Sanders Brown Center on Aging, 3Department of Neurology, University of Kentucky, Lexington, KY 40536, USA. 4Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA. 5Colorado Division of Wildlife, Wildlife Research Center, Fort Collins, CO 80526, USA.

*These authors contributed equally to this work.

†Present address: Department of Infectology, Scripps Research Institute, 5353 Parkside Drive, RF-2, Jupiter, Florida, 33458, USA.

‡Present address: Institute of Neuropathology, University of Zurich, Schmelzbergstrasse 12, 8091 Zurich, Switzerland.

§To whom correspondence should be addressed: E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:gtell2@uky.edu

Prions are transmissible proteinaceous agents of mammals that cause fatal neurodegenerative diseases of the central nervous system (CNS). The presence of infectivity in skeletal muscle of experimentally infected mice raised the possibility that dietary exposure to prions might occur through meat consumption (1). Chronic wasting disease (CWD), an enigmatic and contagious prion disease of North American cervids, is of particular concern. The emergence of CWD in an increasingly wide geographic area and the interspecies transmission of bovine spongiform encephalopathy (BSE) to humans as variant Creutzfeldt Jakob disease (vCJD) have raised concerns about zoonotic transmission of CWD.

To test whether skeletal muscle of diseased cervids contained prion infectivity, Tg(CerPrP)1536 mice (2) expressing cervid prion protein (CerPrP), were inoculated intracerebrally with extracts prepared from the semitendinosus/semimembranosus muscle group of CWD-affected mule deer or from CWD-negative deer. The availability of CNS materials also afforded direct comparisons of prion infectivity in skeletal muscle and brain. All skeletal muscle extracts from CWD-affected deer induced progressive neurological dysfunction in Tg(CerPrP)1536 mice with mean incubation times ranging between 360 and ~490 d, whereas the incubation times of prions from the CNS ranged from ~230 to 280 d (Table 1). For each inoculation group, the diagnosis of prion disease was confirmed by the presence of PrPSc in the brains of multiple infected Tg(CerPrP)1536 mice (see supporting online material for examples). In contrast, skeletal muscle and brain material from CWD-negative deer failed to induce disease in Tg(CerPrP)1536 mice (Table 1) and PrPSc was not detected in the brains of sacrificed asymptomatic mice as late as 523 d after inoculation (supporting online material).

Our results show that skeletal muscle as well as CNS tissue of deer with CWD contains infectious prions. Similar analyses of skeletal muscle BSE-affected cattle did not reveal high levels of prion infectivity (3). It will be important to assess the cellular location of PrPSc in muscle. Notably, while PrPSc has been detected in muscles of scrapie-affected sheep (4), previous studies failed to detect PrPSc by immunohistochemical analysis of skeletal muscle from deer with natural or experimental CWD (5, 6). Since the time of disease onset is inversely proportional to prion dose (7), the longer incubation times of prions from skeletal muscle extracts compared to matched brain samples indicated that prion titers were lower in muscle than in CNS where infectivity titers are known to reach high levels. Although possible effects of CWD strains or strain mixtures on these incubation times cannot be excluded, the variable 360 to ~490 d incubation times suggested a range of prion titers in skeletal muscles of CWD-affected deer. Muscle prion titers at the high end of the range produced the fastest incubation times that were ~30% longer than the incubation times of prions from the CNS of the same animal. Since all mice in each inoculation group developed disease, prion titers in muscle samples producing the longest incubation times were higher than the end point of the bioassay, defined as the infectious dose at which half the inoculated mice develop disease. Studies are in progress to accurately assess prion titers.

While the risk of exposure to CWD infectivity following consumption of prions in muscle is mitigated by relatively inefficient prion transmission via the oral route (8), these

results show that semitendinosus/semimembranosus muscle, which is likely to be consumed by humans, is a significant source of prion infectivity. Humans consuming or handling meat from CWD-infected deer are therefore at risk to prion exposure.

References and Notes

1. P. J. Bosque et al., Proc. Natl. Acad. Sci. U.S.A. 99, 3812 (2002).

2. S. R. Browning et al., J. Virol. 78, 13345 (2004).

3. A. Buschmann, M. H. Groschup, J. Infect. Dis. 192, 934 (2005).

4. O. Andreoletti et al., Nat. Med. 10, 591 (2004).

5. T. R. Spraker et al., Vet. Pathol. 39, 110 (2002).

6. A. N. Hamir, J. M. Miller, R. C. Cutlip, Vet. Pathol. 41, 78 (2004).

7. S. B. Prusiner et al., Biochemistry 21, 4883 (1980).

8. M. Prinz et al., Am. J. Pathol. 162, 1103 (2003).

9. This work was supported by grants from the U.S. Public Health Service 2RO1 NS040334-04 from the National Institute of Neurological Disorders and Stroke and N01-AI-25491 from the National Institute of Allergy and Infectious Diseases.

Supporting Online Material



www.sciencemag.org/



Materials and Methods

Fig. S1

21 November 2005; accepted 13 January 2006 Published online 26 January 2006; 10.1126/science.1122864 Include this information when citing this paper.

Table 1. Incubation times following inoculation of Tg(CerPrP)1536 mice with prions from skeletal muscle and brain samples of CWD-affected deer.

Inocula Incubation time, mean d ± SEM (n/n0)*

Skeletal muscle Brain

CWD-affected deer

H92 360 ± 2 d (6/6) 283 ± 7 d (6/6)

33968 367 ± 9 d (8/8) 278 ± 11 d (6/6)

5941 427 ± 18 d (7/7)

D10 483 ± 8 d (8/8) 231 ± 17 d (7/7)

D08 492 ± 4 d (7/7)

Averages 426 d 264 d

Non-diseased deer

FPS 6.98 >523 d (0/6)

FPS 9.98 >454 d (0/7) >454 d (0/6)

None >490 d (0/6)

PBS >589 d (0/5)

*The number of mice developing prion disease divided by the original number of inoculated mice is shown in parentheses. Mice dying of intercurrent illnesses were excluded.



http://www.sciencemag.org/



www.sciencemag.org/



Supporting Online Material for

Prions in Skeletal Muscles of Deer with Chronic Wasting Disease

Rachel C. Angers, Shawn R. Browning, Tanya S. Seward, Christina J. Sigurdson,

Michael W. Miller, Edward A. Hoover, Glenn C. Telling§

§To whom correspondence should be addressed: E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:gtell2@uky.edu

Published 26 January 2006 on Science Express

DOI: 10.1126/science.1122864

This PDF file includes:

Materials and Methods

Fig. S1

Supporting Online Materials

Materials and Methods

Homogenates of semitendinosus/semimembranosus muscle (10% w/v in phosphate

buffered saline) were prepared from five emaciated and somnolent mule deer, naturally

infected with CWD at the Colorado Division of Wildlife, Wildlife Research Center.

These deer were identified as D10, D08, 33968, H92, and 5941. CWD infection was

confirmed in all cases by the presence of histologic lesions in the brain including

spongiform degeneration of the perikaryon, the immunohistochemical detection of

disease-associated PrP in brain and tonsil, or by immunoblotting of protease-resistant,

disease associated PrP (CerPrPSc). Semitendinosus/semimembranosus muscle was also

obtained from two asymptomatic, mock inoculated deer, referred to as FPS 6.68 and 9.98,

that originated from a CWD non-endemic area and which were held indoors at Colorado

State University from ten days of age. These control deer were confirmed negative for

CWD by histopathological and immunohistochemical analysis of brain tissue at autopsy.

The utmost care was taken to avoid inclusion of obvious nervous tissue when muscle

biopsies were prepared and to ensure that contamination of skeletal muscle samples with

CNS tissue did not occur. Fresh, single-use instruments were used to collect each sample

biopsy and a central piece from each sample was prepared with fresh, disposable

instruments to further isolate muscle tissue for inoculum preparation. Brain samples for

transmission were prepared separately from muscle as additional insurance against cross

contamination.

1

Groups of anesthetized Tg(CerPrP)1536 mice were inoculated intracerebrally with 30 µl

of 1 % skeletal muscle or brain extracts prepared in phosphate buffered saline (PBS).

Inoculated Tg(CerPrP) mice were diagnosed with prion disease following the progressive

development of at least three neurologic symptoms including truncal ataxia, 'plastic' tail,

loss of extensor reflex, difficultly righting, and slowed movement. The time from

inoculation to the onset of clinical signs is referred to as the incubation time.

For PrP analysis in brain extracts of Tg(CerPrP)1536 mice, 10 % homogenates prepared

in PBS were either untreated (-) or treated (+) with 40 µg/ml proteinase K (PK) for one

hour at 37oC in the presence of 2% sarkosyl. Proteins were separated by sodium dodecyl

sulfate polyacrylamide gel electrophoresis, analyzed by immunoblotting using anti PrP

monoclonal antibody 6H4 (Prionics AG, Switzerland), incubated with appropriate

secondary antibody, developed using ECL-plus detection (Amersham), and analyzed

using a FLA-5000 scanner (Fuji).

2

Fig. S1

PrP in brain extracts from representative Tg(CerPrP)1536 mice receiving muscle or CNS

tissue inocula from CWD-affected or CWD-negative deer. Extracts were either treated

(+) or untreated (-) with proteinase K (PK) as indicated. The positions of protein

molecular weight markers at 21.3, 28.7, 33.5 kDa (from bottom to top) are shown to the

left of the immunoblot.

3



http://www.sciencemag.org/



Prions in skeletal muscle



Patrick J. Bosque*,dagger ,Dagger , Chongsuk Ryou*, Glenn Telling*,§, David Peretz*,dagger , Giuseppe Legname*,dagger , Stephen J. DeArmond*,dagger ,¶, and Stanley B. Prusiner*,dagger ,,**

* Institute for Neurodegenerative Diseases and Departments of dagger Neurology, ¶ Pathology, and Biochemistry and Biophysics, University of California, San Francisco, CA 94143

Contributed by Stanley B. Prusiner, December 28, 2001

Considerable evidence argues that consumption of beef products from cattle infected with bovine spongiform encephalopathy (BSE) prions causes new variant Creutzfeldt-Jakob disease. In an effort to prevent new variant Creutzfeldt-Jakob disease, certain "specified offals," including neural and lymphatic tissues, thought to contain high titers of prions have been excluded from foods destined for human consumption [Phillips, N. A., Bridgeman, J. & Ferguson-Smith, M. (2000) in The BSE Inquiry (Stationery Office, London), Vol. 6, pp. 413-451]. Here we report that mouse skeletal muscle can propagate prions and accumulate substantial titers of these pathogens. We found both high prion titers and the disease-causing isoform of the prion protein (PrPSc) in the skeletal muscle of wild-type mice inoculated with either the Me7 o Rocky Mountain Laboratory strain of murine prions. Particular muscles accumulated distinct levels of PrPSc, with the highest levels observed in muscle from the hind limb. To determine whether prions are produced or merely accumulate intramuscularly, we established transgenic mice expressing either mouse or Syrian hamster PrP exclusively in muscle. Inoculating these mice intramuscularly with prions resulted in the formation of high titers of nascent prions in muscle. In contrast, inoculating mice in which PrP expression was targeted to hepatocytes resulted in low prion titers. Our data demonstrate that factors in addition to the amount of PrP expressed determine the tropism of prions for certain tissues. That some muscles are intrinsically capable of accumulating substantial titers of prions is of particular concern. Because significant dietary exposure to prions might occur through the consumption of meat, even if it is largely free of neural and lymphatic tissue, a comprehensive effort to map the distribution of prions in the muscle of infected livestock is needed. Furthermore, muscle may provide a readily biopsied tissue from which to diagnose prion disease in asymptomatic animals and even humans. Dagger Present address: Department of Medicine, Denver Health Medical Center, Denver, CO 80204.

§ Present address: Department of Microbiology and Immunology, University of Kentucky, Lexington, KY 40536-0230.

** To whom reprint requests should be addressed. E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:vann@cgl.ucsf.edu.



http://www.pnas.org/



Extraneural Pathologic Prion Protein in Sporadic Creutzfeldt-Jakob Disease



Markus Glatzel, M.D., Eugenio Abela, Manuela Maissen, M.S., and Adriano Aguzzi, M.D., Ph.D.

snip...

Conclusions Using sensitive techniques, we identified extraneural deposition of PrPSc in spleen and muscle samples from approximately one third of patients who died with sporadic Creutzfeldt-Jakob disease. Extraneural PrPSc appears to correlate with a long duration of disease.



http://content.nejm.org/cgi/content/short/349/19/1812?query=TOC



EMBO reports AOP Published online: 11 April 2003



Widespread PrPSc accumulation in muscles of hamsters orally infected with scrapie



Achim Thomzig, Christine Kratzel, Gudrun Lenz, Dominique KrÒ¼ger & Michael Beekes Robert Koch-Institut, P26, Nordufer 20, D-13353 Berlin, Germany

Received 13 February 2003; Accepted 13 March 2003; Published online 11 April 2003.

Abstract :

Scrapie, bovine spongiform encephalopathy and chronic wasting disease are orally communicable, transmissible spongiform encephalopathies (TSEs). As zoonotic transmissions of TSE agents may pose a risk to human health, the identification of reservoirs for infectivity in animal tissues and their exclusion from human consumption has become a matter of great importance for consumer protection. In this study, a variety of muscles from hamsters that were orally challenged with scrapie was screened for the presence of a molecular marker for TSE infection, PrPSc (the pathological isoform of the prion protein PrP). Sensitive western blotting revealed consistent PrPSc accumulation in skeletal muscles from forelimb and hindlimb, head, back and shoulder, and in tongue. Previously, our animal model has provided substantial baseline information about the peripheral routing of infection in naturally occurring and orally acquired ruminant TSEs. Therefore, the findings described here highlight further the necessity to investigate thoroughly whether muscles of TSE-infected sheep, cattle, elk and deer contain infectious agents.



http://www.emboreports.org/



Detection of Prion Infectivity in Fat Tissues of Scrapie-Infected Mice

Brent Race1#, Kimberly Meade-White1#, Michael B. A. Oldstone2, Richard Race1, Bruce Chesebro1*

1 Laboratory of Persistent Virus Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, United States of America, 2 Department of Immunology and Microbial Science, The Scripps Research Institute, LaJolla, California, United States of America

Abstract Distribution of prion infectivity in organs and tissues is important in understanding prion disease pathogenesis and designing strategies to prevent prion infection in animals and humans. Transmission of prion disease from cattle to humans resulted in banning human consumption of ruminant nervous system and certain other tissues. In the present study, we surveyed tissue distribution of prion infectivity in mice with prion disease. We show for the first time detection of infectivity in white and brown fat. Since high amounts of ruminant fat are consumed by humans and also incorporated into animal feed, fat-containing tissues may pose a previously unappreciated hazard for spread of prion infection.

Author Summary Prion diseases, also known as transmissible spongiform encephalopathies, are infectious progressive fatal neurodegenerative diseases which affect humans as well as wild and domestic animals. Distribution of prion infectivity in organs and tissues is important in understanding prion disease pathogenesis and designing strategies to prevent prion infection in animals and humans. We show for the first time the presence of prion infectivity in white fat and brown fat tissues of mice with prion disease. Our results suggest that fat tissues of domestic or wild animals infected with prions may pose an unappreciated hazard for spread of infection to humans or domestic animals. The presence of prion infectivity in fat suggests that additional consideration may be required to eliminate from the food chain any fat from ruminants suspected of exposure to or infection with prions. Thus, this finding has implications for public health, food safety, and prion disease prevention strategies.

Citation: Race B, Meade-White K, Oldstone MBA, Race R, Chesebro B (2008) Detection of Prion Infectivity in Fat Tissues of Scrapie-Infected Mice. PLoS Pathog 4(12): e1000232. doi:10.1371/journal.ppat.1000232

Editor: Neil Mabbott, University of Edinburgh, United Kingdom

Received: August 12, 2008; Accepted: November 5, 2008; Published: December 5, 2008

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

Funding: This research was supported in part by the Intramural Research Program of the NIH, NIAID. MBAO was funded through NIA grant AG04032.

Competing interests: The authors have declared that no competing interests exist.

* E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:bchesebro@nih.gov

# These authors contributed equally to this work.

Introduction

snip...

Discussion The present results indicate that white fat and brown fat are possible tissue sources of prion infectivity which might play a role in transmission of prion disease. In vivo brown fat has a limited distribution, usually found in young animals in the intrascapular region and around various organs such as heart and kidney. In adult ruminants brown fat is minimal. Therefore, brown fat from infected animals is unlikely to be consumed by humans in large amounts. In contrast, humans often consume large amounts of ruminant white fat. In premium cuts of meat containing mostly skeletal muscle, white fat is often intertwined with muscle cells, and it is impossible to separate the two cell types. However, white fat, free of muscle, is found in subcutaneous, retroperitoneal, intraperitoneal, perirenal and other regions. Such fat is used in many processed meat products such as sausages and canned meats, and is also used in animal feeds. Our present data show clearly that fat in the absence of muscle has significant infectivity titers, which are similar to titers in muscle containing fat (Table 1). Since our skeletal muscle samples are unavoidably contaminated by white fat, it is possible that fat might be a contributor to the infectivity found in muscle. In support of this possibility we found PrPres detectable by IHC at high levels in white fat associated with skeletal muscle in some tg44 mice (Figure 4). In contrast, other groups did not mention seeing PrPres in muscle-associated fat tissue in animals where myocytes themselves were seen to be positive by IHC [13]-[20].

snip...

It is unclear why there is accumulation of PrPres and infectivity in adipose tissues. One possibility might be the high level of innervation by the autonomic nervous system in both brown and white fat. In WT mice, nerves should express cell membrane anchored PrPC (PrPsen). Sympathetic nerves have been previously implicated in transfer of scrapie infectivity from spleen to brain in mice [29], and they might also play a role in infection of fat in WT mice. In tg44 mice the mechanism of fat infection is likely to be different as there is no anchored PrPsen on the nerves. We currently postulate a role for connective tissue structures in this process.

Infectivity in fat might also contribute to environmental contamination following the death of prion infected animals. Although infectivity titers are lower in fat and muscle than in CNS, the large mass of fat and muscle makes the total infectivity from these sources similar. Furthermore, fat and muscle are readily accessible to the environment after death, whereas the CNS is highly confined in skull and vertebral column. These factors might increase the importance of fat and muscle as sources of spread of prion disease among animals.

The low or negative plasma titers found in tg44 and WT mice indicate that residual plasma cannot account for the high infectivity levels seen in fat and other tissues (Table 1). However, low levels of plasma or blood-borne infectivity might still be a mechanism for spread of infectivity among tissues in tg44 mice and possibly also WT mice. Similarly transmission of low level blood prion infectivity has been documented by blood transfusion in BSE-infected sheep [30], and also accounts for some rare cases of human variant CJD [31],[32].

In this study extraneural infection was much higher in tg44 mice expressing anchorless PrP than in WT mice. The explanation of this finding is unclear. Possibly soluble anchorless PrP facilitates spread of infection from CNS to extraneural sites by blood, lymph or nerve-mediated transport. Alternatively, the long asymptomatic survival time of tg44 mice might also contribute to high level extraneural infection. This could also be a factor in many animal prion diseases where the time course is long, i.e. 2-5 years or more, and might allow higher extraneural infectivity in fat tissues [7], [33]-[35].

The present data using a mouse model shows the proof of principle that brown and white fat tissues can be important sites of prion agent deposition. It will be important to extend these studies in the future to prion infected large animals such as cattle, sheep and cervids where there may be greater potential for contamination of human or domestic animal food chains. We are currently doing this experiment with fat from CWD deer; however, it will require an additional year to gather this data, and this result is therefore beyond the scope of the present paper. Such studies may be difficult because of the lower titers seen in these large animals compared to rodent scrapie models. For example, we often detect titers of 9-10 logID50/gram of mouse brain, whereas in brain from BSE cattle [8], and scrapie sheep [4] titers reported are 7-8 logID50/gram. We are finding similar low titers in CWD cervid brain in our deer PrP transgenic mice (unpublished data). These results could indicate either that the amount of prion agent present in ruminant brain is lower than in mice and hamsters or that the cattle, sheep and deer PrP transgenic mice used for infectivity assays are less sensitive than the WT mice or hamster PrP transgenic mice used for rodent scrapie. In either case this might affect ability to detect infectivity in fat of these important large animal models.

Materials and Methods

snip...full text ;



http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1000232



Research Project: Study of Atypical Bse Location: Virus and Prion Diseases of Livestock


Project Number: 3625-32000-086-05 Project Type: Specific Cooperative Agreement

Start Date: Sep 15, 2004 End Date: Sep 14, 2009

Objective: The objective of this cooperative research project with Dr. Maria Caramelli from the Italian BSE Reference Laboratory in Turin, Italy, is to conduct comparative studies with the U.S. bovine spongiform encephalopathy (BSE) isolate and the atypical BSE isolates identified in Italy. The studies will cover the following areas: 1. Evaluation of present diagnostics tools used in the U.S. for the detection of atypical BSE cases. 2. Molecular comparison of the U.S. BSE isolate and other typical BSE isolates with atypical BSE cases. 3. Studies on transmissibility and tissue distribution of atypical BSE isolates in cattle and other species.

Approach: This project will be done as a Specific Cooperative Agreement with the Italian BSE Reference Laboratory, Istituto Zooprofilattico Sperimentale del Piemonte, in Turin, Italy. It is essential for the U.S. BSE surveillance program to analyze the effectiveness of the U.S diagnostic tools for detection of atypical cases of BSE. Molecular comparisons of the U.S. BSE isolate with atypical BSE isolates will provide further characterization of the U.S. BSE isolate. Transmission studies are already underway using brain homogenates from atypical BSE cases into mice, cattle and sheep. It will be critical to see whether the atypical BSE isolates behave similarly to typical BSE isolates in terms of transmissibility and disease pathogenesis. If transmission occurs, tissue distribution comparisons will be made between cattle infected with the atypical BSE isolate and the U.S. BSE isolate. Differences in tissue distribution could require new regulations regarding specific risk material (SRM) removal.



http://www.ars.usda.gov/research/projects/projects.htm?accn_no=408490



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

From: Terry S. Singeltary Sr. To: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000248/!x-usc:mailto:justin.greenlee@ars.usda.gov

Sent: Monday, June 15, 2009 4:30 PM

Subject: re-Research Project: Study of Atypical Bse


re-Research Project: Study of Atypical Bse



http://www.ars.usda.gov/research/projects/projects.htm?ACCN_NO=408490



Greetings Sir,

I have been most interested in these transmission studies, and i know it is probably to early for you to say much about them. but, could you please give me an update of some kind, as to if transmission of any type occured, with any tissues and or body fluids. i suppose i am a bit anxious about these studies. ...

many thanks for your work,

kindest regards, terry



=======end...tss======



???



Monday, June 01, 2009

Biochemical typing of pathological prion protein in aging cattle with BSE



http://bse-atypical.blogspot.com/2009/06/biochemical-typing-of-pathological.html



Sunday, June 07, 2009

L-TYPE-BSE, H-TYPE-BSE, C-TYPE-BSE, IBNC-TYPE-BSE, TME, CWD, SCRAPIE, CJD, NORTH AMERICA



http://bse-atypical.blogspot.com/2009/06/l-type-bse-h-type-bse-c-type-bse-ibnc.html



Sunday, May 10, 2009

Identification and characterization of bovine spongiform encephalopathy cases diagnosed and NOT diagnosed in the United States



http://bse-atypical.blogspot.com/2009/05/identification-and-characterization-of.html



Sunday, December 28, 2008

MAD COW DISEASE USA DECEMBER 28, 2008 an 8 year review of a failed and flawed policy



http://bse-atypical.blogspot.com/2008/12/mad-cow-disease-usa-december-28-2008-8.html



Wednesday, August 20, 2008

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



http://bse-atypical.blogspot.com/2008/08/bovine-spongiform-encephalopathy-mad.html



Saturday, February 28, 2009

NEW RESULTS ON IDIOPATHIC BRAINSTEM NEURONAL CHROMATOLYSIS "All of the 15 cattle tested showed that the brains had abnormally accumulated PrP" 2009 SEAC 102/2



http://bse-atypical.blogspot.com/2009/02/new-results-on-idiopathic-brainstem.html



Saturday, June 13, 2009

BSE FEED VIOLATIONS USA UPDATE From 01/01/2009 To 06/10/2009



http://madcowfeed.blogspot.com/2009/06/bse-feed-violations-usa-update-from.html



Thursday, March 19, 2009

MILLIONS AND MILLIONS OF POUNDS OF MAD COW FEED IN COMMERCE USA



http://madcowfeed.blogspot.com/2009/03/millions-and-millions-of-pounds-of-mad.html



WHO WILL FOLLOW THE CHILDREN FOR CJD SYMPTOMS ???



Saturday, May 2, 2009

U.S. GOVERNMENT SUES WESTLAND/HALLMARK MEAT OVER USDA CERTIFIED DEADSTOCK DOWNER COW SCHOOL LUNCH PROGRAM



http://downercattle.blogspot.com/2009/05/us-government-sues-westlandhallmark.html



Sunday, April 12, 2009

BSE MAD COW TESTING USA 2009 FIGURES Month Number of Tests

Feb 2009 -- 1,891

Jan 2009 -- 4,620



http://www.aphis.usda.gov/newsroom/hot_issues/bse/surveillance/ongoing_surv_results.shtml



SEE FULL TEXT ;



http://madcowtesting.blogspot.com/2009/04/bse-mad-cow-testing-usa-2009-figures.html



Monday, May 4, 2009

Back to the Past With New TSE Testing Agricultural Research/May-June 2009



http://madcowtesting.blogspot.com/2009/05/back-to-past-with-new-tse-testing.html



Thursday, April 9, 2009

Docket No. FDA2002N0031 (formerly Docket No. 2002N0273) RIN 0910AF46 Substances Prohibited From Use in Animal Food or Feed; Final Rule: Proposed



http://madcowfeed.blogspot.com/2009/04/docket-no-fda2002n0031-formerly-docket.html



http://prionunitusaupdate2008.blogspot.com/2009/04/r-calf-and-usa-mad-cow-problem-dont.html#comments



Sunday, April 12, 2009

r-calf and the USA mad cow problem, don't look, don't find, and then blame Canada



http://prionunitusaupdate2008.blogspot.com/2009/04/r-calf-and-usa-mad-cow-problem-dont.html



http://prionunitusaupdate2008.blogspot.com/2009/04/cjd-foundation-sides-with-r-calfers-no.html#comments



Sunday, May 10, 2009

Meeting of the Transmissible Spongiform Encephalopathies Committee On June 12, 2009 (Singeltary submission)



http://tseac.blogspot.com/2009/05/meeting-of-transmissible-spongiform.html



Saturday, June 13, 2009

Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States 2003 revisited 2009

snip...

2006

FOOD AND DRUG ADMINISTRATION

DEPARTMENT OF HEALTH AND HUMAN SERVICES

This transcripts has not been edited or corrected, but appears as received from the commercial transcribing service. Accordingly, the food and Drug Administration makes no representation as to its accuracy.

Meeting of:

TRANSMISSIBLE

SPONGIFORM ENCEPHALOPATHIES

ADVISORY COMMITTEE

September 18, 2006

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.



http://www.fda.gov/ohrms/dockets/ac/06/transcripts/1006-4240t1.htm



http://www.fda.gov/ohrms/dockets/ac/06/transcripts/2006-4240t1.pdf



2008

I ask Professor Kong ;

Thursday, December 04, 2008 3:37 PM Subject: RE: re--Chronic Wating Disease (CWD) and Bovine Spongiform Encephalopathies (BSE): Public Health Risk Assessment

''IS the h-BSE more virulent than typical BSE as well, or the same as cBSE, or less virulent than cBSE? just curious.....''

Professor Kong reply ;

.....snip

''As to the H-BSE, we do not have sufficient data to say one way or another, but we have found that H-BSE can infect humans. I hope we could publish these data once the study is complete. Thanks for your interest.''

Best regards, Qingzhong Kong, PhD Associate Professor Department of Pathology Case Western Reserve University Cleveland, OH 44106 USA

END...TSS

I look forward to further transmission studies, and a true ENHANCED BSE/atypical BSE surveillance program put forth testing all cattle for human and animal consumption for 5 years. a surveillance program that uses the most sensitive TSE testing, and has the personnel that knows how to use them, and can be trusted. I look forward to a stringent mad cow feed ban being put forth, and then strictly enforced. we need a forced, not voluntary feed ban, an enhanced feed ban at that, especially excluding blood. we need some sort of animal traceability. no more excuses about privacy. if somebody is putting out a product that is killing folks and or has the potential to kill you, then everybody needs to know who they are, and where that product came from. same with hospitals, i think medical incidents in all states should be recorded, and made public, when it comes to something like a potential accidental transmission exposure event. so if someone is out there looking at a place to go have surgery done, if you have several hospitals having these type 'accidental exposure events', than you can go some place else. it only makes sense. somewhere along the road, the consumer lost control, and just had to take whatever they were given, and then charged these astronomical prices. some where along the line the consumer just lost interest, especially on a long incubating disease such as mad cow disease i.e. Transmissible Spongiform Encephalopathy. like i said before, there is much more to the mad cow story than bovines and eating a hamburger, we must start focusing on all TSE in all species. ...TSS



http://bse-atypical.blogspot.com/2009/02/atypical-bse-north-america-update.html



snip... SEE FULL TEXT ;





Sunday, May 10, 2009



Meeting of the Transmissible Spongiform Encephalopathies Committee On June 12, 2009 (Singeltary submission)



http://cjdusa.blogspot.com/2009/06/monitoring-occurrence-of-emerging-forms.html



ALL Human and Animal Transmissible Spongiform Encephalopathy, of all phenotypes, of ALL ages, in EVERY State and INTERNATIONALLY, should be made MANDATORY reportable ASAP. ...



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

L-TYPE-BSE, H-TYPE-BSE, C-TYPE-BSE, IBNC-TYPE-BSE, TME, CWD, SCRAPIE, CJD, NORTH AMERICA

L-TYPE-BSE, H-TYPE-BSE, C-TYPE-BSE, IBNC-TYPE-BSE, TME, CWD, SCRAPIE, CJD, NORTH AMERICA, something to ponder ;

confusious ask, what if ;


Session I - Prions: Structure, Strain and Detection (II)

Searching for BASE Strain Signature in Sporadic Creutzfedlt-Jakob Disease

Gianluigi Zanusso

Department of Neurological and Visual Sciences, Section of Clinical Neurology University of Verona, Verona, Italy.

Bovine amyloidotic spongiform encephalopathy (BASE) is a newly recognized form of bovine prion disease, which was originally detected in Italy in 2004 as an effect of active surveillance. BASE or BSE L-type (L is referred to the lower electrophoretic PrPSc migration than classical BSE) has now been reported in several countries, including Japan. All field cases of BASE were older than 8 years and neurologically normal at the time of slaughtered. By experimental transmission, we defined the disease phenotype of cattle BASE, which is quite distinct from that seen in typical BSE and characterized by mental dullness and amyotrophy. Surprisingly, following intraspecies and interspecies transmission the incubation period of BASE was shorter than BSE. The relatively easy transmission of BASE isolate as well as the molecular similarity with sporadic Creutzfeldt-Jakob disease (sCJD) have raised concern regarding its potential passage to humans. Tg humanized mice Met/Met at codon 129 challenged with both BSE and BASE isolates, showed a resistance to BSE but a susceptibility to BASE at a 60% rate; in addition, BASE-inoculated Cynomolgus (129 Met/Met) had shorter incubation periods than BSE-inoculated primates. In this study we compared the biochemical properties of PrPSc in Cynomolgus and in TgHu Met/Met mice challenged with BSE and BASE strains, by conventional SDS-PAGE analysis and 2D separation. The results obtained disclose distinct conformational changes in PrPSc, which are dependent on the inoculated host but not on the codon 129 genotype.

This work was supported by Neuroprion contract n. FOOD CT 2004 -506579 (NOE)



http://www.istitutoveneto.it/prion_09/Abstracts_09.pdf




P26

TRANSMISSION OF ATYPICAL BOVINE SPONGIFORM ENCEPHALOPATHY (BSE) IN HUMANIZED MOUSE MODELS

Liuting Qing1, Fusong Chen1, Michael Payne1, Wenquan Zou1, Cristina Casalone2, Martin Groschup3, Miroslaw Polak4, Maria Caramelli2, Pierluigi Gambetti1, Juergen Richt5*, and Qingzhong Kong1

1Department of Pathology, Case Western Reserve University, Cleveland, OH 44106, USA; 2CEA, Istituto Zooprofilattico Sperimentale, Italy; 3Friedrich-Loeffler-Institut, Germany; 4National Veterinary Research Institute, Poland; 5Kansas State University, Diagnostic Medicine/Pathobiology Department, Manhattan, KS 66506, USA. *Previous address: USDA National Animal Disease Center, Ames, IA 50010, USA

Classical BSE is a world-wide prion disease in cattle, and the classical BSE strain (BSE-C) has led to over 200 cases of clinical human infection (variant CJD). Two atypical BSE strains, BSE-L (also named BASE) and BSE-H, have been discovered in three continents since 2004. The first case of naturally occurring BSE with mutated bovine PrP gene (termed BSE-M) was also found in 2006 in the USA. The transmissibility and phenotypes of these atypical BSE strains/isolates in humans were unknown.

We have inoculated humanized transgenic mice with classical and atypical BSE strains (BSE-C, BSE-L, BSE-H) and the BSE-M isolate. We have found that the atypical BSE-L strain is much more virulent than the classical BSE-C. The atypical BSE-H strain is also transmissible in the humanized transgenic mice with distinct phenotype, but no transmission has been observed for the BSE-M isolate so far.



http://www.istitutoveneto.it/prion_09/Abstracts_09.pdf



Wednesday, February 11, 2009

Atypical BSE North America Update February 2009 Both of the BSE cases ascertained in the US native-born cattle were atypical cases (H-type), which contributed to the initial ambiguity of the diagnosis. 174, 185 In Canada, there have been 2 atypical BSE cases in addition to the 14 cases of the classic UK strain of BSE2: one was the H-type, and the other was of the L-type.198


snip...end


source : Enhanced Abstract Journal of the American Veterinary Medical Association January 1, 2009, Vol. 234, No. 1, Pages 59-72 Bovine spongiform encephalopathy Jane L. Harman, DVM, PhD; Christopher J. Silva, PhD



http://avmajournals.avma.org/doi/ref/10.2460/javma.234.1.59




Atypical BSE North America Update February 2009



http://bse-atypical.blogspot.com/2009/02/atypical-bse-north-america-update.html



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

Emmanuel E. Comoy1*, Cristina Casalone2, Nathalie Lescoutra-Etchegaray1, Gianluigi Zanusso3, Sophie Freire1, Dominique Marcé1, Frédéric Auvré1, Marie-Magdeleine Ruchoux1, Sergio Ferrari3, Salvatore Monaco3, Nicole Salès4, Maria Caramelli2, Philippe Leboulch1,5, Paul Brown1, Corinne I. Lasmézas4, Jean-Philippe Deslys1

1 Institute of Emerging Diseases and Innovative Therapies, CEA, Fontenay-aux-Roses, France, 2 Istituto Zooprofilattico Sperimentale del Piemonte, Turin, Italy, 3 Policlinico G.B. Rossi, Verona, Italy, 4 Scripps Florida, Jupiter, Florida, United States of America, 5 Genetics Division, Brigham & Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America

Abstract Top Background Human variant Creutzfeldt-Jakob Disease (vCJD) results from foodborne transmission of prions from slaughtered cattle with classical Bovine Spongiform Encephalopathy (cBSE). Atypical forms of BSE, which remain mostly asymptomatic in aging cattle, were recently identified at slaughterhouses throughout Europe and North America, raising a question about human susceptibility to these new prion strains.

Methodology/Principal Findings Brain homogenates from cattle with classical BSE and atypical (BASE) infections were inoculated intracerebrally into cynomolgus monkeys (Macacca fascicularis), a non-human primate model previously demonstrated to be susceptible to the original strain of cBSE. The resulting diseases were compared in terms of clinical signs, histology and biochemistry of the abnormal prion protein (PrPres). The single monkey infected with BASE had a shorter survival, and a different clinical evolution, histopathology, and prion protein (PrPres) pattern than was observed for either classical BSE or vCJD-inoculated animals. Also, the biochemical signature of PrPres in the BASE-inoculated animal was found to have a higher proteinase K sensitivity of the octa-repeat region. We found the same biochemical signature in three of four human patients with sporadic CJD and an MM type 2 PrP genotype who lived in the same country as the infected bovine.

Conclusion/Significance Our results point to a possibly higher degree of pathogenicity of BASE than classical BSE in primates and also raise a question about a possible link to one uncommon subset of cases of apparently sporadic CJD. Thus, despite the waning epidemic of classical BSE, the occurrence of atypical strains should temper the urge to relax measures currently in place to protect public health from accidental contamination by BSE-contaminated products.

Citation: Comoy EE, Casalone C, Lescoutra-Etchegaray N, Zanusso G, Freire S, et al. (2008) Atypical BSE (BASE) Transmitted from Asymptomatic Aging Cattle to a Primate. PLoS ONE 3(8): e3017. doi:10.1371/journal.pone.0003017

Editor: Neil Mabbott, University of Edinburgh, United Kingdom

Received: April 24, 2008; Accepted: August 1, 2008; Published: August 20, 2008

Copyright: © 2008 Comoy et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: This work has been supported by the Network of Excellence NeuroPrion.

Competing interests: CEA owns a patent covering the BSE diagnostic tests commercialized by the company Bio-Rad.

* E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000110/!x-usc:mailto:emmanuel.comoy@cea.fr



http://www.plosone.org/article/info:doi/10.1371/journal.pone.0003017




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 and L-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

Abstract

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

snip...

Conclusion

These studies provide experimental evidence that the Stetsonville TME agent is distinct from typical BSE but has phenotypic similarities to L-type BSE in TgOvPrP4 mice. Our conclusion is that L-type BSE is a more likely candidate for a bovine source of TME infection than typical BSE. In the scenario that a ruminant TSE is the source for TME infection in mink, this would be a second example of transmission of a TSE from ruminants to non-ruminants under natural conditions or farming practices in addition to transmission 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 mice and macaques, suggests that L-type BSE is more pathogenic for humans than typical BSE (24,38).



http://www.cdc.gov/eid/content/13/12/1887.htm?s_cid=eid1887_e




Transmissible Mink Encephalopathy TME

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




http://transmissible-mink-encephalopathy.blogspot.com/2007/12/phenotypic-similarity-of-transmissible.html




Dr. Thornsberry: Let's hypothesize that I had some cattle on the eastern slope and they were in the same pasture with elk with CWD. If a cow had been exposed to the PRP Scrapie and it did develop disease four years later, would that look like BSE? Would there be a way to determine if it came from CWD?

Dr. Bartz: The IC studies in cattle indicate it does not look like BSE. The clinical signs of the IC/CWD cattle are more like downer cattle, and not aggressive. As far as finding the source of a bovine TSE, the gold standard is the lesion profile study where you take cattle tissue and inoculate it into mice with appropriate controls, wait until the mice come down, and do the lesion profiling.




http://transmissible-mink-encephalopathy.blogspot.com/



http://transmissible-mink-encephalopathy.blogspot.com/2006_12_01_archive.html




3.57 The experiment which might have determined whether BSE and scrapie were caused by the same agent (ie, the feeding of natural scrapie to cattle) was never undertaken in the UK. It was, however, performed in the USA in 1979, when it was shown that cattle inoculated with the scrapie agent endemic in the flock of Suffolk sheep at the United States Department of Agriculture in Mission, Texas, developed a TSE quite unlike BSE.339 The findings of the initial transmission, though not of the clinical or neurohistological examination, were communicated in October 1988 to Dr Watson, Director of the CVL, following a visit by Dr Wrathall, one of the project leaders in the Pathology Department of the CVL, to the United States Department of Agriculture.340 The results were not published at this point, since the attempted transmission to mice from the experimental cow brain had been inconclusive. The results of the clinical and histological differences between scrapie-affected sheep and cattle were published in 1995. Similar studies in which cattle were inoculated intracerebrally with scrapie inocula derived from a number of scrapie-affected sheep of different breeds and from different States, were carried out at the US National Animal Disease Centre.341 The results, published in 1994, showed that this source of scrapie agent, though pathogenic for cattle,

*** did not produce the same clinical signs of brain lesions characteristic of BSE. ***

3.58 There are several possible reasons why the experiment was not performed in the UK. It had been recommended by Sir Richard Southwood (Chairman of the Working Party on Bovine Spongiform Encephalopathy) in his letter to the Permanent Secretary of MAFF, Mr (now Sir) Derek Andrews, on 21 June 1988,342 though it was not specifically recommended in the Working Party Report or indeed in the Tyrrell Committee Report (details of the Southwood Working Party and the Tyrell Committee can be found in vol. 4: The Southwood Working Party, 1988–89 and vol. 11: Scientists after Southwood respectively). The direct inoculation of scrapie into calves was given low priority, because of its high cost and because it was known that it had already taken place in the USA.343 It was also felt that the results of such an experiment would be hard to interpret. While a negative result 337 Fraser, H., Bruce, M., Chree, A., McConnell, I. and Wells, G. (1992) Transmission of Bovine Spongiform Encephalopathy and Scrapie to Mice, Journal of General Virology, 73, 1891–7; Bruce, M., Chree, A., McConnell, I., Foster, J., Pearson, G. and Fraser, H. (1994) Transmission of Bovine Spongiform Encephalopathy and Scrapie to Mice: Strain Variation and the Species Barrier, Philosophical Transactions of the Royal Society of London, Series B, Biological Sciences, 343, 405–11 338 Bruce, M., Will, R., Ironside, J., McConell, I., Drummond, D., Suttie, A., McCordie, L., Chree, A., Hope, J., Birkett, C., Cousens, S., Fraser, H. and Bostock, C. (1997) Transmissions to Mice Indicate that ‘New Variant’ CJD is Caused by the BSE Agent, Nature, 389, 498–501 339 Clark, W., Hourrigan, J. and Hadlow, W. (1995) Encephalopathy in Cattle Experimentally Infected with the Scrapie Agent, American Journal of Veterinary Research, 56, 606–12 340 YB88/10.00/1.1 341 Cutlip, R., Miller, J., Race, R., Jenny, A., Katz, J., Lehmkuhl, H., Debey, B. and Robinson, M. (1994) Intracerebral Transmission of Scrapie to Cattle, Journal of Infectious Diseases, 169, 814–20 342 YB88/6.21/1.2 343 YB88/11.17/2.4 SCIENCE 84 would be informative, a positive result would need to demonstrate that when scrapie was transmitted to cattle, the disease which developed in cattle was the same as BSE.344 Given the large number of strains of scrapie and the possibility that BSE was one of them, it would be necessary to transmit every scrapie strain to cattle separately, to test the hypothesis properly. Such an experiment would be expensive. Secondly, as measures to control the epidemic took hold, the need for the experiment from the policy viewpoint was not considered so urgent. It was felt that the results would be mainly of academic interest.345 3.59 Nevertheless, from the first demonstration of transmissibility of BSE in 1988, the possibility of differences in the transmission properties of BSE and scrapie was clear. Scrapie was transmissible to hamsters, but by 1988 attempts to transmit BSE to hamsters had failed. Subsequent findings increased that possibility.



http://www.bseinquiry.gov.uk/pdf/volume2/chapter3.pdf




Monday, May 11, 2009

Rare BSE mutation raises concerns over risks to public health



http://bse-atypical.blogspot.com/2009/05/rare-bse-mutation-raises-concerns-over.html




Sunday, April 12, 2009 BSE MAD COW TESTING USA 2009 FIGURES Month Number of Tests

Feb 2009 -- 1,891

Jan 2009 -- 4,620



http://www.aphis.usda.gov/newsroom/hot_issues/bse/surveillance/ongoing_surv_results.shtml



SEE FULL TEXT ;



http://madcowtesting.blogspot.com/2009/04/bse-mad-cow-testing-usa-2009-figures.html




Monday, May 4, 2009

Back to the Past With New TSE Testing Agricultural Research/May-June 2009



http://madcowtesting.blogspot.com/2009/05/back-to-past-with-new-tse-testing.html




Sunday, May 10, 2009

Identification and characterization of bovine spongiform encephalopathy cases diagnosed and NOT diagnosed in the United States



http://bse-atypical.blogspot.com/2009/05/identification-and-characterization-of.html



Saturday, February 28, 2009NEW RESULTS ON IDIOPATHIC BRAINSTEM NEURONAL CHROMATOLYSIS TYPE BSE

"All of the 15 cattle tested showed that the brains had abnormally accumulated PrP" 2009
SEAC 102/2


http://bse-atypical.blogspot.com/2009/02/new-results-on-idiopathic-brainstem.html



Wednesday, October 08, 2008

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



http://bse-atypical.blogspot.com/2008/10/idiopathic-brainstem-neuronal.html




''THE LINE TO TAKE'' ON IBNC $$$ 1995 $$$

1995

page 9 of 14 ;30. The Committee noted that the results were unusual. the questioned whether there could be coincidental BSE infection or contamination with scrapie. Dr. Tyrell noted that the feeling of the committee was that this did not represent a new agent but it was important to be prepared to say something publicly about these findings. A suggested line to take was that these were scientifically unpublishable results but in line with the policy of openness they would be made publicly available and further work done to test their validity. Since the BSE precautions were applied to IBNC cases, human health was protected. Further investigations should be carried out on isolations from brains of IBNC cases with removal of the brain and subsequent handling under strict conditions to avoid the risk of any contamination.31. Mr. Bradley informed the Committee that the CVO had informed the CMO about the IBNC results and the transmission from retina and he, like the Committee was satisfied that the controls already in place or proposed were adequate. ...

snip... see full text


http://www.bseinquiry.gov.uk/files/yb/1995/06/21005001.pdf



Wednesday, August 20, 2008

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

August 20, 2008


snip...


another question, just how long have these atypical BSE TSEs been around in the bovine ???let's look at another case of atypical BSE in Germany way back in 1992 ;Subject: atypical BSE reported in 1992 and conviently slaughterd and incinerated and then swept under rug for about 12 years Date: April 26, 2007 at 1:08 pm PST 1992NEW BRAIN DISORDER3. WHAT ABOUT REPORTS OF NEW FORM OF BSE?THE VETERINARY RECORD HAS PUBLISHED AN ARTICLE ON A NEW BRAIN DISORDER OF CATTLE DISCOVERED THROUGH OUR CONTROL MEASURES FOR BSE. ALTHOUGH IT PRESENTS SIMILAR CLINICAL SIGNS TO BSE THERE ARE MAJOR DIFFERENCES IN THE HISTOPATHOLOGY AND INCUBATION PERIODS BETWEEN THE TWO. MUST EMPHASISE THAT THIS IS NOT BSE.4. IS THIS NEW BRAIN DISORDER A THREAT?WE DO NOT EVEN KNOW WHETHER THE AGENT OF THIS DISEASE IS TRANSMISSIBLE. IN ANY CASE, CASES SO FAR IDENTIFIED HAD SHOWN SIMILAR SYMPTOMS TO THOSE OF BSE, AND THEREFORE HAVE BEEN SLAUGHTERED AND INCINERATED, SO THAT IF A TRANSMISSIBLE AGENT WERE INVOLVED IT WOULD HAVE BEEN ELIMINATED. .......



http://www.bseinquiry.gov.uk/files/yb/1992/10/26001001.pdf




2. The Collinge/Will dispute appears to rumble on. Dr. Collinge had told Dr. Tyrrell that Dr. Will's response to his criticism about sharing material had been ''quite unacceptable'' (in spite of it's apparently conciliatory tone). Apparently Professor Allen was now going to try and arrange a meeting to resolve the dispute. No action here for MAFF, although Mr. Murray may be interested.3. Dr. Tyrrell regretted that the Committee had not seen the article on BBD. However he felt that for the time being NO specific action was called for. The most important need was to consider the possibility that the condition might be transmissible. As we have discussed, I suggested that we might circulate a paper to the members of the committee giving our appreciation of this condition (and perhaps of other non-BSE neurological conditions that had been identified in negative cases) and of any necessary follow up action. IF any Committee member felt strongly about this, or if the issue CAME TO A HEAD, we would call an interim meeting. He was happy with this approach. I would be grateful if Mr. Maslin could, in discussion with CVL and veterinary colleagues draft such a note, which will presumably very largely follow what Mr. Bradley's briefing paper has already said, taking account of DOH comments, We can then clear a final version with DOH before circulating it to Committee members.




http://www.bseinquiry.gov.uk/files/yb/1992/10/29005001.pdf




IN CONFIDENCE

This is a highly competitive field and it really will be a pity if we allow many of the key findings to be published by overseas groups while we are unable to pursue our research findings because of this disagreement, which I hope we can make every effort to solve.




http://www.bseinquiry.gov.uk/files/yb/1992/10/26002001.pdf




COLLINGE THREATENS TO GO TO MEDIA




http://www.bseinquiry.gov.uk/files/yb/1992/12/16005001.pdf




2. The discovery might indicate the existence of a different strain of BSE from that present in the general epidemic or an unusual response by an individual host.3. If further atypical lesion distribution cases are revealed in this herd then implications of misdiagnosis of 'negative' cases in other herds may not be insignificant.snip...This minute is re-issued with a wider distribution. The information contained herein should NOT be disseminated further except on the basis of ''NEED TO KNOW''.

R Bradley


http://www.bseinquiry.gov.uk/files/yb/1993/02/17001001.pdf



IN CONFIDENCE

BSE ATYPICAL LESION DISTRIBUTION


http://www.bseinquiry.gov.uk/files/yb/1993/03/14001001.pdf



ALABAMA MAD COW CASE



snip...


see full text ;



http://bse-atypical.blogspot.com/2008/08/bovine-spongiform-encephalopathy-mad.html



Friday, May 29, 2009

Characterization of a U.S. Sheep Scrapie Isolate with Short Incubation Time


http://scrapie-usa.blogspot.com/2009/05/characterization-of-us-sheep-scrapie.html


Friday, May 29, 2009

Seven Deer Test Positive for Chronic Wasting Disease During 2009 Spring Collections in Hampshire County, West Virginia



http://chronic-wasting-disease.blogspot.com/2009/05/seven-deer-test-positive-for-chronic.html



O.K. confusious asks, IF all these new atypical BSEs i.e. new strains of mad cow disease is just an 'OLD COW PRION DISEASE', why then can not the 'old human prion disease' such as the sporadic CJD, be from an 'old cow prion disease', same as the nvCJD 'young people mad cow disease' (which also happens in 74 year old), but why cannot the 'old cow prion diseases', i.e. l-BSE, h-BSE, and ibncBSE, cause the 'old people prion disease', which looks like sporadic CJD. seems that is what some of the pathology is showing ???

OH, that probably makes too much sense, and that the only answer could be that it's all just a happenstance of bad luck and or a spontaneous event, that just happens out of the clear blue sky $$$

IF this is the case, then where are all the SPONTANEOUS BSE CASES OF MAD COW DISEASE IN THE U.S.A., AND WHERE HAVE THEY BEEN BURIED IN THE USA OVER THE LAST 25 YEARS ???


Thursday, April 30, 2009

FDA Issues Final Guidance for Renderers on Substances Prohibited From Use in Animal Food or Feed CVM Update Back April 30, 2009



http://madcowfeed.blogspot.com/2009/04/fda-issues-final-guidance-for-renderers.html




Sunday, December 28, 2008

MAD COW DISEASE USA DECEMBER 28, 2008 an 8 year review of a failed and flawed policy



http://bse-atypical.blogspot.com/2008/12/mad-cow-disease-usa-december-28-2008-8.html




Wednesday, August 20, 2008

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



http://bse-atypical.blogspot.com/2008/08/bovine-spongiform-encephalopathy-mad.html





Monday, June 01, 2009

Biochemical typing of pathological prion protein in aging cattle with BSE



http://bse-atypical.blogspot.com/2009/06/biochemical-typing-of-pathological.html




SPORADIC CJD CASES RISING IN U.S.A


Monday, April 20, 2009 National Prion Disease Pathology Surveillance Center Cases Examined1 (December 31, 2008)

April 20, 2009

National Prion Disease Pathology Surveillance Center Cases Examined1 (December 31, 2008)

National Prion Disease Pathology Surveillance Center Cases Examined1

(December 31, 2008)

Year Total Referrals2 Prion Disease Sporadic Familial Iatrogenic vCJD

1996 & earlier 42 32 28 4 0 0

1997 115 68 59 9 0 0

1998 93 53 45 7 1 0

1999 115 69 61 8 0 0

2000 151 103 89 14 0 0

2001 210 118 108 9 0 0

2002 258 147 123 22 2 0

2003 273 176 135 41 0 0

2004 335 184 162 21 0 13

2005 346 193 154 38 1 0

2006 380 192 159 32 0 14

2007 370 212 185 26 0 0

2008 383 228 182 23 0 0

TOTAL 30715 17756 1490 254 4 2

1 Listed based on the year of death or, if not available, on year of referral; 2 Cases with suspected prion disease for which brain tissue and/or blood (in familial cases) were submitted; 3 Disease acquired in the United Kingdom; 4 Disease acquired in Saudi Arabia; 5 Includes 20 cases in which the diagnosis is pending, and 17 inconclusive cases; 6 Includes 25 cases with type determination pending in which the diagnosis of vCJD has been excluded.

Rev 2/13/09 National



http://www.cjdsurveillance.com/pdf/case-table.pdf



http://www.cjdsurveillance.com/resources-casereport.html



http://www.aan.com/news/?event=read&article_id=4397&page=72.45.45



*5 Includes 20 cases in which the diagnosis is pending, and 17 inconclusive cases; *6 Includes 25 cases with type determination pending in which the diagnosis of vCJD has been excluded.


Greetings,


it would be interesting to know what year these atypical cases occurred, as opposed to lumping them in with the totals only.

are they accumulating ?

did they occur in one year, two years, same state, same city ?

location would be very interesting ?

age group ?

sex ?

how was it determined that nvCJD was ruled out ?

from 1997, the year i started dealing with this nightmare, there were 28 cases (per this report), up until 2007 where the total was 185 cases (per this report), and to date 2008 is at 182. a staggering increase in my opinion, for something that just happens spontaneously as some would have us believe. i don't believe it, not in 85%+ of all sporadic CJD cases. actually, i do not believe yet that anyone has proven that any of the sporadic CJD cases have been proven to be a spontaneous misfolding of a protein. there are many potential routes and sources for the sporadic CJD's. ...TSS

please see full text here ;



http://prionunitusaupdate2008.blogspot.com/2009/04/national-prion-disease-pathology.html




Monday, June 01, 2009

Biochemical typing of pathological prion protein in aging cattle with BSE


http://bse-atypical.blogspot.com/2009/06/biochemical-typing-of-pathological.html



Friday, May 29, 2009

Characterization of a U.S. Sheep Scrapie Isolate with Short Incubation Time


http://scrapie-usa.blogspot.com/2009/05/characterization-of-us-sheep-scrapie.html




Friday, May 29, 2009

Seven Deer Test Positive for Chronic Wasting Disease During 2009 Spring Collections in Hampshire County, West Virginia



http://chronic-wasting-disease.blogspot.com/2009/05/seven-deer-test-positive-for-chronic.html




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

Biochemical typing of pathological prion protein in aging cattle with BSE

Biochemical typing of pathological prion protein in aging cattle with BSE

Virology Journal 2009, 6:64 doi:10.1186/1743-422X-6-64 Seraina Tester (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:seraina.tester@itn.unibe.ch) Valerie Juillerat (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:valerie.juillerat@itn.unibe.ch) Marcus G Doherr (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:marcus.doherr@itn.unibe.ch) Bianca Haase (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:bianca.haase@itz.unibe.ch) Miroslaw Polak (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:ppolak@piwet.pulawy.pl) Felix Ehrensperger (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:f.ehrensperger@vetpath.uzh.ch) Tosso Leeb (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:tosso.leeb@itz.unibe.ch) Andreas Zurbriggen (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:andreas.zurbriggen@itn.unibe.ch) Torsten Seuberlich (mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:torsten.seuberlich@itn.unibe.ch) ISSN 1743-422X Article type Research Submission date 23 March 2009 Acceptance date 26 May 2009 Publication date 26 May 2009 Article URL http://www.virologyj.com/content/6/1/64 This peer-reviewed article was published immediately upon acceptance. It can be downloaded, printed and distributed freely for any purposes (see copyright notice below). Articles in Virology Journal are listed in PubMed and archived at PubMed Central. For information about publishing your research in Virology Journal or any BioMed Central journal, go to http://www.virologyj.com/info/instructions/ For information about other BioMed Central publications go to http://www.biomedcentral.com/ Virology Journal © 2009 Tester et al. , licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 1

Biochemical typing of pathological prion protein in aging cattle with BSE

Seraina Tester1, Valerie Juillerat1, Marcus G. Doherr1, Bianca Haase2, Miroslaw Polak3, Felix Ehrensperger4, Tosso Leeb2, Andreas Zurbriggen1 and Torsten Seuberlich1* 1 NeuroCenter, Reference Laboratory for TSE in animals, Department of Clinical Research and Veterinary Public Health, Vetsuisse Faculty, University of Berne, Switzerland 2 Institute of Genetics, Vetsuisse Faculty, University of Berne, Switzerland 3 National Veterinary Research Institute, Pulawy, Poland 4 Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zürich, Switzerland ST: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:seraina.tester@itn.unibe.ch VJ: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:vallerie.juillerat@itn.unibe.ch MGD: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:marcus.doherr@itn.unibe.ch BH: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:bianca.haase@itz.unibe.ch MP: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:ppolak@piwet.pulawy.pl FE: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:f.ehrensperger@vetpath.uzh.ch TL: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:tosso.leeb@itz.unibe.ch AZ: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:andreas.zurbriggen@itn.unibe.ch *Corresponding author: Dr. med. vet. Torsten Seuberlich NeuroCenter - Reference Laboratory for TSE in animals Department of Clinical Research and VPH Vetsuisse Faculty University of Berne Bremgartenstrasse 109a CH- 3001 Berne Switzerland Email: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:torsten.seuberlich@itn.unibe.ch Phone: +41 31 631 2206 Fax: +41 31 631 2538 2

Abstract:

Background

The broad enforcement of active surveillance for bovine spongiform encephalopathy (BSE) in 2000 led to the discovery of previously unnoticed, atypical BSE phenotypes in aged cattle that differed from classical BSE (C-type) in biochemical properties of the pathological prion protein. Depending on the molecular mass and the degree of glycosylation of its proteinase K resistant core fragment (PrPres), mainly determined in samples derived from the medulla oblongata, these atypical cases are currently classified into low (L)-type or high (H)-type BSE. In the present study we address the question to what extent such atypical BSE cases are part of the BSE epidemic in Switzerland.

Results

To this end we analyzed the biochemical PrPres type by Western blot in a total of 33 BSE cases in cattle with a minimum age of eight years, targeting up to ten different brain regions. Our work confirmed H-type BSE in a zebu but classified all other cases as C-type BSE; indicating a very low incidence of H- and L-type BSE in Switzerland. It was documented for the first time that the biochemical PrPres type was consistent across different brain regions of aging animals with C-type and H-type BSE, i.e. independent of the neuroanatomical structure investigated.

Conclusions

Taken together this study provides further characteristics of the BSE epidemic in Switzerland and generates new baseline data for the definition of C- and H-type BSE phenotypes, thereby underpinning the notion that they indeed represent distinct prion disease entities.

snip...

Conclusions

Taken together these results indicate that the prevalence of H- and L-type BSE in Switzerland remains under the detection limit of the Swiss active surveillance program. However one H-type BSE case was identified by passive BSE surveillance and proves in principle the capacity to identify such cases in the population. Hence, the overall prevalence of atypical BSE in Switzerland appears very low and similar to what has been reported from other countries. It has been speculated and strengthened by experimental data [53,54] that atypical BSE once recycled in the cattle population was the origin of the worldwide BSE epidemic in the last 20 years. If this holds true and such cases occur spontaneously in the population, then BSE might never be completely eradicated. Furthermore, in these circumstances, it would be hazardous to relieve certain disease control measures, including the total prohibition of MBM in ruminant feed.



http://www.virologyj.com/content/pdf/1743-422X-6-64.pdf





Greetings,


O.K., let me get this straight, we have typical, U.K. c-BSE, we have now a spontaneous atypical h-BSE and l-BSE, of which they are now just calling an 'old cow disease', which happens spontaneously without any route or source, just happens.

OH, and then we have the new/old ibncBSE, of which is just another 'old cow prion disease', another one of those spontaneous events $$$

IT SEEMS there is a pattern here to make all Transmissible Spongiform Encephalopathies a spontaneous event, even though there is no such evidence what so ever.

WITH all the sub-types of TSE showing up now in the Scrapie, BSE, CWD, (all of which has been rendered and fed back to livestock producing animals for human and animal feed), and even in humans i.e. CJD, there is no way, they know for a fact that all are of a spontaneous, sporadic event, that just happened due to a twisted up protein that folded the wrong way by itself, from no route and no source. they do NOT know if any of these atypical TSE are of a spontaneous old cows prion disease or not, but yet they preach it like it's the gospel $$$

I don't believe them today, and i will never believe that 85%+ of all human sporadic CJD, just happens without any route and source from anything. ...


kind regards, terry



Absence of spontaneous disease and comparative prion susceptibility of transgenic mice expressing mutant human prion proteins

Emmanuel A. Asante, Ian Gowland, Andrew Grimshaw, Jacqueline M. Linehan, Michelle Smidak, Richard Houghton, Olufunmilayo Osiguwa, Andrew Tomlinson, Susan Joiner, Sebastian Brandner, Jonathan D. F. Wadsworth and John Collinge Correspondence John Collinge mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:j.collinge@prion.ucl.ac.uk MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology, National Hospital for Neurology and Neurosurgery, Queen Square, London WC1N 3BG, UK Received 15 October 2008 Accepted 2 December 2008

Approximately 15% of human prion disease is associated with autosomal-dominant pathogenic mutations in the prion protein (PrP) gene. Previous attempts to model these diseases in mice have expressed human PrP mutations in murine PrP, but this may have different structural consequences. Here, we describe transgenic mice expressing human PrP with P102L or E200K mutations and methionine (M) at the polymorphic residue 129. Although no spontaneous disease developed in aged animals, these mice were readily susceptible to prion infection from patients with the homotypic pathogenic mutation. However, while variant Creutzfeldt-Jakob disease (CJD) prions transmitted infection efficiently to both lines of mice, markedly different susceptibilities to classical (sporadic and iatrogenic) CJD prions were observed. Prions from E200K and classical CJD M129 homozygous patients, transmitted disease with equivalent efficiencies and short incubation periods in human PrP 200K, 129M transgenic mice. However, mismatch at residue 129 between inoculum and host dramatically increased the incubation period. In human PrP 102L, 129M transgenic mice, short disease incubation periods were only observed with transmissions of prions from P102L patients, whereas classical CJD prions showed prolonged and variable incubation periods irrespective of the codon 129 genotype. Analysis of diseaserelated PrP (PrPSc) showed marked alteration in the PrPSc glycoform ratio propagated after transmission of classical CJD prions, consistent with the PrP point mutations directly influencing PrPSc assembly. These data indicate that P102L or E200K mutations of human PrP have differing effects on prion propagation that depend upon prion strain type and can be significantly influenced by mismatch at the polymorphic residue 129.

snip...

Transgenic mice expressing high levels of mouse PrP 101L (equivalent to 102L in human PrP) spontaneously developed neurological dysfunction at 166 days of age (Hsiao et al., 1990). PrPSc levels were low or undetectable, and brain extracts from affected mice did not transmit CNS degeneration to wild-type mice, but transmission to hamsters and Tg(GSSPrP)196 mice, expressing lower levels of the same mutant transgene product, was reported (Hsiao et al., 1994; Telling et al., 1996a). These Tg(GSSPrP)196 mice have subsequently been reported to develop spontaneous disease at advanced age (Tremblay et al., 2004; Nazor et al., 2005). It therefore remains debateable as to whether prions had been generated in these transgenic mice or this simply represents acceleration of a spontaneous neurodegenerative disease already poised to occur in these mice (Nazor et al., 2005). Others generated transgenic mice expressing endogenous levels of mouse PrP 101L by the gene knock-in approach (Manson et al., 1999). These mice did not develop spontaneous neurodegeneration but were reported to show greater susceptibility to human P102L prions than wildtype mice (Barron et al., 2001).

However, we consider it essential to study this and other human pathogenic mutations in human PrP, rather than in mouse PrP where the mutation may have different structural consequences. With respect to such models it is important to demonstrate that human PrP is functionally active and can participate in prion propagation and pathogenesis in mouse cells. Human PrP can rescue a PrP null phenotype in mice (Whittington et al., 1995), confirming it is functionally active and human prions can replicate in transgenic mice expressing only human PrP, which develop spongiform neurodegeneration (Collinge et al., 1995).

snip...see full text ;



http://vir.sgmjournals.org/cgi/reprint/90/3/546



****** [2] Correction Date: Tue 5 Sep 2006 From: "Terry S Singeltary Sr" <mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:flounder9@verizon.net>

Characterization of atypical BSE in Germany:


correction -------------------------------------------------------


[In the Moderator's comment accompanying the abstract of the paper entitled "Atypical BSE in Germany-Proof of transmissibility and biochemical characterization'" by A Buschmannaet et al, (see part [2] of CJD (new var.) update 2006 (09) 20060904.2519) it was wrongly implied that Terry S Singeltary Sr endorsed the conclusions of the paper, whereas his comments were intended merely to highlight the conclusions of the paper. Namely that the atypical cases suggested the possible existence of sporadic BSE cases in bovines and perhaps the BSE epidemic in the UK could have also been initiated by an intraspecies transmission from a sporadic BSE case. I apologize for inadvertently misrepresenting Terry's views. - Mod.CP]

Terry S Singeltary Sr has written the following. "In fact I disagree with the spontaneous/sporadic BSE/TSE theory, IF this is what the authors of this paper meant by 'sporadic BSE' to mean. For one thing, it has never been proven. IF atypical BSE i.e. BASE is so similar to some sporadic CJDs, then how did they all of a sudden become spontaneous? Could it not be so simple as an atypical BSE i.e. BASE was transmitted the same way most of all of the other BSE cattle were i.e. feed of just an atypical source, thus causing atypical strain? Why would these animals not develop an atypical BSE i.e. BASE from the same oral route? WHAT about an atypical strain mutating to become infectious via a lateral or horizontal mode in the bovine, as with CWD and scrapie? Also, please explain to me how a distinct synthetic prion, of a strain that is supposedly unlike any other we have ever seen, how can this explain 6 different documented phenotypes of sporadic CJD to date?

It's like trying to explain away all the 6 phenotypes of sporadic CJD with the spontaneous theory, it's just not scientific. OR, if you render an atypical TSE of what ever phenotype, in what ever species, of the atypical strain and feed it to another whatever species, nothing happens x 1 x 2 x 3 x 4 etc passage? This all has been proven?

Please show me these transmission studies? What Prusiner and Soto produced in vitro did not look like any natural field TSE, and as far as this in vitro TSE being infectious, well this was questionable too. If this was the case, then why does CWD not spontaneously happen in geographical areas where it has never been documented, OR with scrapie, as in scrapie free New Zealand? If TSE were to arise spontaneously, I don't see how the scientific arena can dictate which animal TSE can arise spontaneously, and which ones cannot, without any scientific evidence to support this to date, and by even suggesting this in this study, was not scientific. The words sporadic and spontaneous are very confusing in the world literature of human and animal TSE and, in my opinion, should not be used as terminology of any TSE."

-- Terry S Singeltary Sr <mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000056/!x-usc:mailto:flounder9@verizon.net>



http://apex.oracle.com/pls/otn/f?p=2400:1202:17822992441545446841::NO::F2400_P1202_CHECK_DISPLAY,F2400_P1202_PUB_MAIL_ID:X,34659




Saturday, February 28, 2009

NEW RESULTS ON IDIOPATHIC BRAINSTEM NEURONAL CHROMATOLYSIS "All of the 15 cattle tested showed that the brains had abnormally accumulated PrP" 2009 SEAC 102/2



http://bse-atypical.blogspot.com/2009/02/new-results-on-idiopathic-brainstem.html




Sunday, May 17, 2009

De Novo Generation of Infectious Prions In Vitro Produces a New Disease Phenotype



http://bse-atypical.blogspot.com/2009/05/de-novo-generation-of-infectious-prions.html




Monday, May 11, 2009

Rare BSE mutation raises concerns over risks to public health



http://bse-atypical.blogspot.com/2009/05/rare-bse-mutation-raises-concerns-over.html





Sunday, April 12, 2009


BSE MAD COW TESTING USA 2009 FIGURES Month Number of Tests


Feb 2009 -- 1,891

Jan 2009 -- 4,620



http://www.aphis.usda.gov/newsroom/hot_issues/bse/surveillance/ongoing_surv_results.shtml




SEE FULL TEXT ;



http://madcowtesting.blogspot.com/2009/04/bse-mad-cow-testing-usa-2009-figures.html




Monday, May 4, 2009

Back to the Past With New TSE Testing Agricultural Research/May-June 2009



http://madcowtesting.blogspot.com/2009/05/back-to-past-with-new-tse-testing.html




Sunday, May 10, 2009

Identification and characterization of bovine spongiform encephalopathy cases diagnosed and NOT diagnosed in the United States



http://bse-atypical.blogspot.com/2009/05/identification-and-characterization-of.html




Friday, May 29, 2009

Characterization of a U.S. Sheep Scrapie Isolate with Short Incubation Time




http://scrapie-usa.blogspot.com/2009/05/characterization-of-us-sheep-scrapie.html




Friday, May 29, 2009

Seven Deer Test Positive for Chronic Wasting Disease During 2009 Spring Collections in Hampshire County, West Virginia




http://chronic-wasting-disease.blogspot.com/2009/05/seven-deer-test-positive-for-chronic.html




O.K. confusious asks, IF all these new atypical BSEs i.e. new strains of mad cow disease is just an 'OLD COW PRION DISEASE', why then can not the 'old human prion disease' such as the sporadic CJD, be from an 'old cow prion disease', same as the nvCJD 'young people mad cow disease' (which also happens in 74 year old), but why cannot the 'old cow prion diseases', i.e. l-BSE, h-BSE, and ibncBSE, cause the 'old people prion disease', which looks like sporadic CJD. seems that is what some of the pathology is showing ???


OH, that probably makes too much sense, and that the only answer could be that it's all just a happenstance of bad luck and or a spontaneous event, that just happens out of the clear blue sky $$$


IF this is the case, then where are all the SPONTANEOUS BSE CASES OF MAD COW DISEASE IN THE U.S.A., AND WHERE HAVE THEY BEEN BURIED IN THE USA OVER THE LAST 25 YEARS ???


Atypical BSE North America Update February 2009



http://bse-atypical.blogspot.com/2009/02/atypical-bse-north-america-update.html




Sunday, December 28, 2008

MAD COW DISEASE USA DECEMBER 28, 2008 an 8 year review of a failed and flawed policy



http://bse-atypical.blogspot.com/2008/12/mad-cow-disease-usa-december-28-2008-8.html




Wednesday, August 20, 2008

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



http://bse-atypical.blogspot.com/2008/08/bovine-spongiform-encephalopathy-mad.html





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

De Novo Generation of Infectious Prions In Vitro Produces a New Disease Phenotype

Sent: Saturday, May 16, 2009 9:49 PM

Subject: De Novo Generation of Infectious Prions In Vitro Produces a New Disease Phenotype

De Novo Generation of Infectious Prions In Vitro Produces a New Disease Phenotype

Marcelo A. Barria1,2, Abhisek Mukherjee1,2, Dennisse Gonzalez-Romero1,2, Rodrigo Morales1,2,3, Claudio Soto1,2*

1 George and Cynthia Mitchell Center for Neurodegenerative Diseases, University of Texas Medical Branch, Galveston, Texas, United States of America, 2 Department of Neurology, University of Texas Houston Medical School, Houston, Texas, United States of America, 3 Facultad de Ciencias, University of Chile, Santiago, Chile

Abstract

Prions are the proteinaceous infectious agents responsible for Transmissible Spongiform Encephalopathies. Compelling evidence supports the hypothesis that prions are composed exclusively of a misfolded version of the prion protein (PrPSc) that replicates in the body in the absence of nucleic acids by inducing the misfolding of the cellular prion protein (PrPC). The most common form of human prion disease is sporadic, which appears to have its origin in a low frequency event of spontaneous misfolding to generate the first PrPSc particle that then propagates as in the infectious form of the disease. The main goal of this study was to mimic an early event in the etiology of sporadic disease by attempting de novo generation of infectious PrPSc in vitro. For this purpose we analyzed in detail the possibility of spontaneous generation of PrPSc by the protein misfolding cyclic amplification (PMCA) procedure. Under standard PMCA conditions, and taking precautions to avoid cross-contamination, de novo generation of PrPSc was never observed, supporting the use of the technology for diagnostic applications. However, we report that PMCA can be modified to generate PrPSc in the absence of pre-existing PrPSc in different animal species at a low and variable rate. De novo generated PrPSc was infectious when inoculated into wild type hamsters, producing a new disease phenotype with unique clinical, neuropathological and biochemical features. Our results represent additional evidence in support of the prion hypothesis and provide a simple model to study the mechanism of sporadic prion disease. The findings also suggest that prion diversity is not restricted to those currently known, and that likely new forms of infectious protein foldings may be produced, resulting in novel disease phenotypes.

Citation: Barria MA, Mukherjee A, Gonzalez-Romero D, Morales R, Soto C (2009) De Novo Generation of Infectious Prions In Vitro Produces a New Disease Phenotype. PLoS Pathog 5(5): e1000421. doi:10.1371/journal.ppat.1000421 Editor: David Westaway, University of Alberta, Canada Received April 7, 2008; Accepted April 9, 2009; Published May 15, 2009 Copyright:
2009 Barria et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by grants from the National Institute of Neurological Disorders (R01 NS49173), the National Institute of Allergy and Infectious Diseases (P01 AI77774) and the National Institute of Aging (P01 AG14359). None of the sponsors or funders played any role in the design and conduct of the study, in the collection, analysis, and interpretation of the data, or in the preparation, review, or approval of the manuscript. Competing Interests: Dr. Soto is an inventor on the PMCA technology and a Founder, Vice-President and Chief Scientific Officer of Amprion Inc, a biotech company focusing on the development of early and sensitive diagnosis for prion diseases and other disorders involving protein misfolding. * E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000223/!x-usc:mailto:claudio.soto@uth.tmc.edu

Discussion

In this study we report spontaneous generation of PrPSc in vitro using a modified PMCA procedure with brain homogenate substrate. Misfolded protein was produced in two different animal species without the need to add seeds of in vivo generated PrPSc. At present it is unclear whether PMCA replicated a PrPSc intermediate already present in the brain homogenate or rather ‘‘spontaneous’’ misfolding was induced by the amplification cycles. In this context, a recent study reported the presence of small quantities of a PrPSc-like protein in healthy brains [20], but it is yet unknown whether this protein is infectious.

Although de novo prion formation was achieved before by PMCA using purified proteins in the presence of synthetic polyanions [4], the purpose of our study was to evaluate whether de novo generation of prions is possible under standard PMCA conditions. This is important to assess the validity of using PMCA for prion diagnosis. Another aim was to study whether different species of animals have a distinct propensity to form spontaneous prions under a given set of conditions. Finally, we aimed to characterize the newly generated infectivity and to show that the diversity of prions is perhaps larger than we currently think. Our results indicate that standard PMCA rounds consisting of 144 cycles (or less) did not show spontaneous formation of PrPSc, but longer rounds of 240 cycles were successful in replicating or generating misfolded protein. These findings suggest that de novo formation of PrPSc can be experimentally distinguished from replication of pre-formed PrPSc, indicating that the biochemical, conformational or stability properties of the PrP structures involved in both processes are probably different. Therefore, PMCA can be adapted to produce de novo generated PrPSc and it is likely that more drastic modifications of the procedure may lead to higher rates of spontaneous prion formation. Indeed, the higher rate of de novo PrPSc formation reported in the study by Deleault and colleagues, who used only 48 PMCA cycles per round [4], indicates that using polyanions and purified PrPC may favor spontaneous prion formation, suggesting that some factors in brain homogenate may prevent PrP misfolding.

Our data support the use of PMCA, (under conditions in which PrPSc de novo generation does not occur), for biochemical detection of prions and its potential application for TSE diagnosis. The usefulness of PMCA for highly sensitive and specific detection of prions in biological fluids, has been demonstrated by studies from us and other groups [21–25].

Strikingly, the rate and number of PMCA cycles required to produce de novo PrPSc was variable in distinct species. Considering that from the species studied humans are the only one in which sporadic disease is known to occur, we were surprised that de novo PrPSc was never obtained with human brain homogenates under the experimental conditions used. This contrast with the high levels of amplification of human PrPSc observed in samples seeded with vCJD (Fig. S1) or various sCJD brain homogenates (data not shown). The absence of spontaneous formation of PrPres in humans may be due to the different conditions for the preparation of the tissue, including several hours of post-mortem delay or the lack of perfusion before collection of the brain. However, this is unlikely, because similar results were obtained using brains of transgenic mice expressing human PrP subjected to the same conditions to prepare brain than the other rodent species. Our interpretation of these results is that human PrPC has a lower propensity to initiate misfolding than the rodent protein. The alternative explanation that factors present in the human brain may prevent conversion is unlikely considering the experiments with humanized transgenic mice. The appearance of sporadic disease in humans may simply reflect the longer life span that provides greater chances for stochastic processes of spontaneous misfolding. However, it is also possible that sporadic disease is actually more frequent in animals, because the rate of spontaneous illness in animals has not been systematically studied.

Several lines of evidence enable us to rule out the possibility that de novo PrPSc was the result of cross-contamination. First of all, conventional PMCA that allows us to amplify as little as a single particle of PrPSc did not show spontaneous formation of misfolded protein, even after extensive PMCA cycling (Fig. 1). Second, the rate of spontaneous generation of prions in different species does not correlate with the availability of infectious material in our lab. Moreover, the variable rate was surprisingly constant among different experiments, arguing that it is dependent on some intrinsic properties of the protein rather than a stochastic event as cross-contamination. Third, an experiment done using strictly prion-free equipment and reagents, performed in a new laboratory that was never exposed to prions and done by a person who has never been in contact with prion material showed very similar results as the one carried out in our standard facilities. Finally, the biochemical, structural and biological properties of de novo generated PrPSc in hamster were substantially different to those of various conventional hamster prion strains. Although all these evidences strongly indicate that the results are not due to crosscontamination, this possibility cannot entirely be ruled out.

Inoculation of wild type hamsters with de novo generated PrPSc produced disease in all animals. Strikingly, many of the disease characteristics were substantially different to those observed with several other hamster prion strains. In this study we directly compared PGP-h1 with 3 well-established strains (263K, HY and DY) and literature comparisons with other strains such as Sc237, SHa(Me7), MT-C5, SHa(RML), 139H and Me7-H, also show substantial differences [26,27]. It is important to note that PMCA amplification of PrPSc from various strains (of hamsters, mice, human or deer origin) faithfully propagates the strain characteristics [8–10,28,29]. Thus, the differences observed in the de novo generated material are not attributable to PMCA amplification, but to intrinsic differences of this new prion strain. In our direct comparisons the incubation time of PGP-h1 was significantly larger than 263K and HY strains, but much shorter than DY. Clinical signs were also clearly distinguishable from those observed in animals affected by other strains (see Video S1 and Video S2 in supporting online material). The animals inoculated with PGP-h1 were not hyperactive or aggressive, but not lethargic either. They exhibited a social withdrawal and lack of interest for the surroundings, which reflected in a much reduced horizontal and vertical activity and increased extent of inactive time in an open field test. The clinical differences on the PGP-h1 induced disease were likely the result of the severe brain damage produced, including extensive spongiform degeneration, PrPSc accumulation and brain inflammation. Remarkably, the brain areas targeted by vacuolation damage were significantly different from all other hamster strains studied and included a substantially greater extent of spongiosis in colliculus and a much reduced level of injury in cerebellum and cortex. The biochemical characteristics of PrPSc were also different in PGP-h1, mostly in terms of the higher quantity of PrPSc accumulated in the brain and the lower relative resistance to proteolytic degradation than the other hamster strains analyzed. Taken together this data demonstrate that de novo generated prions correspond to a novel strain of infectious material, able to generate a new disease in wild type animals. We are currently assessing the infectious properties of some of the other de novo generated PrPSc obtained in this study to examine whether or not new strains and new diseases are produced also in other species.

Our findings provide a model to study the possible origins of sporadic TSEs and a new avenue to investigate the mechanism and factors controlling spontaneous formation of infectious material. For example, using the modified PMCA reaction described in this study we could assess the sequence determinants of the variable propensity of PrPC in distinct species to undergo conversion into the pathological form. We could also study the contribution of other factors in brain to alter the rate of de novo generation of PrPSc. Finally, our data provides further support for the prion hypothesis, since misfolded and infectious protein was generated in vitro, without the need for addition of pre-existing PrPSc. The fact that the de novo generated PrPSc corresponds to a new strain of infectious material suggest that the diversity of alternative and transmissible foldings that PrPSc can adopt is much larger than usually thought. This is worrisome, because it raises the possibility that novel and perhaps more aggressive infectious prion foldings may spontaneously originate in diverse species, leading to the emergence of new and unpredictable forms of transmissible diseases.


http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=2675078&blobtype=pdf



Subject: USDA, SPONTANEOUS MAD COW DISEASE, THE TOOTH FAIRY AND SANTA CLAUS

Date: June 12, 2006 at 5:18 am PST

IF we all believe the BSe that the USDA is trying to put out now about atypical BSE in USA cattle just arising spontaneously, then we all should believe in the tooth fairy and santa claus as well.

IF USA scrapie transmitted to USA cattle long ago in experiments in a lab in Mission Texas did not produce UK BSE, but something very different, then why would USA TSE cattle produce the UK human version of mad cow i.e. nvCJD? IT wouldn't. USA sporadic cjd is increasing, the USA also has atypical human cases of unknown origin as well?

THERE are over 20 strains of scrapie, plus the atypical in sheep, and these strains are increasing in numbers.

SCRAPIE, CWD, AND TSE IN CATTLE i.e. ANIMAL TSE RAMPANT IN USA FOR DECADES, and amplified via rendering and feeding practices, where USDA triple firewalls against BSE were nothing more than a mere smoke screen.

NO test tube TSE by either Prusiner or Soto, to date, have ever produced a TSE identical to the sporadic CJD. IN fact, no test tube TSE has ever been produced that resembles _any_ natural field TSE.

IF you feed BSE tainted materials to cattle and primate, you have BSE and nvCJD. IF you feed USA sheep strain to USA cattle, you get USA TSE. IF you feed USA tainted cattle to humans, you get USA mad cow disease. IF you feed sporadic CJD to primate you get a CJD infected primate. NOTHING spontaneous about it at all.

USA is in a very unique situation. there are more documented TSE in different species than any other country, all of which have been rendered and fed back to animals for human and animal consumption, for decades. Millions exposed, and of these Millions, how many surgical and dental procedures have been done on these exposed, to pass on to others, via the 'friendly fire' mode of transmission?

IF, the spontaneous TSE was true, then this would be Prusiner and everyone else that is trying to cash in on this agent with there TSE rapid test, this would be there dream come true. IT would require mandatory BSE/TSE testing of all species, due to the fact you could not ever eradicate it through any intervention. BUT, then again, the spontaneous TSE is like believing in the tooth fairy or santa clause will be arriving at your house this year.

How long can this sharade continue $

How many more will become exposed and have to die $

snip...


http://www.prwatch.org/node/4883



http://scienceblogs.com/aetiology/2006/06/emerging_disease_and_zoonoses_8.php



DEADLY FEAST RICHARD RHODES 1997

EVEN the origin of sporadic CJD continues to be debated. A British researcher, Dr. Richard Kimberlin, points out that the age-specific incidence of sporadic CJD: is similar to the age-specific incidence of BSE.

snip...

Kimberlin argues that the bell curve of sporadic CJD indicates that this seemingly random human disease is probably also caused by infection: "The shape of the age-specific incidence curve... implies that infection with [a] common strain [of CJD] occurs in childhood or adolescence, and that the median incubation period is 40 to 50 years."

German researcher Dr. Heino Diringer similarly defends an infectious cause: "It seems more than likely that ... the sporadic cases of CJD always originate from direct or indirect transmission from animals to man."

END

1: EMBO J. 2002 Dec 2;21(23):6358-66. Links BSE prions propagate as either variant CJD-like or sporadic CJD-like prion strains in transgenic mice expressing human prion protein.Asante EA, Linehan JM, Desbruslais M, Joiner S, Gowland I, Wood AL, Welch J, Hill AF, Lloyd SE, Wadsworth JD, Collinge J. MRC Prion Unit, Institute of Neurology, University College, Queen Square, London WC1N 3BG, UK.

Variant Creutzfeldt-Jakob disease (vCJD) has been recognized to date only in individuals homozygous for methionine at PRNP codon 129. Here we show that transgenic mice expressing human PrP methionine 129, inoculated with either bovine spongiform encephalopathy (BSE) or variant CJD prions, may develop the neuropathological and molecular phenotype of vCJD, consistent with these diseases being caused by the same prion strain. 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 PrP(Sc) type 2. These data suggest that more than one BSE-derived 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.


http://www.ncbi.nlm.nih.gov/pubmed/12456643



http://www.neurology.org/cgi/eletters/60/2/176



2009

Monday, May 11, 2009

Rare BSE mutation raises concerns over risks to public health


http://bse-atypical.blogspot.com/2009/05/rare-bse-mutation-raises-concerns-over.html



Rare BSE mutation raises concerns over risks to public health

SIR - Atypical forms (known as H- and L-type) of bovine spongiform encephalopathy (BSE) have recently appeared in several European countries as well as in Japan, Canada and the United States. This raises the unwelcome possibility that variant Creutzfeldt-Jakob disease (vCJD) could increase in the human population. Of the atypical BSE cases tested so far, a mutation in the prion protein gene (PRNP) has been detected in just one, a cow in Alabama with BSE; her healthy calf also carried the mutation (J. A. Richt and S. M. Hall PLoS Pathog. 4, e1000156; 2008). This raises the possibility that the disease could occasionally be genetic in origin. Indeed, the report of the UK BSE Inquiry in 2000 suggested that the UK epidemic had most likely originated from such a mutation and argued against the scrapierelated assumption. Such rare potential pathogenic PRNP mutations could occur in countries at present considered to be free of BSE, such as Australia and New Zealand. So it is important to maintain strict surveillance for BSE in cattle, with rigorous enforcement of the ruminant feed ban (many countries still feed ruminant proteins to pigs). Removal of specified risk material, such as brain and spinal cord, from cattle at slaughter prevents infected material from entering the human food chain. Routine genetic screening of cattle for PRNP mutations, which is now available, could provide additional data on the risk to the public. Because the point mutation identified in the Alabama animals is identical to that responsible for the commonest type of familial (genetic) CJD in humans, it is possible that the resulting infective prion protein might cross the bovine-human species barrier more easily. Patients with vCJD continue to be identified. The fact that this is happening less often should not lead to relaxation of the controls necessary to prevent future outbreaks. Malcolm A. Ferguson-Smith Cambridge University Department of Veterinary Medicine, Madingley Road, Cambridge CB3 0ES, UK e-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000223/!x-usc:mailto:maf12@cam.ac.uk Jürgen A. Richt College of Veterinary Medicine, Kansas State University, K224B Mosier Hall, Manhattan, Kansas 66506-5601, USA

NATUREVol 45726 February 2009


http://www.nature.com/nature/journal/v457/n7233/full/4571079b.html



Sunday, May 10, 2009

Identification and characterization of bovine spongiform encephalopathy cases diagnosed and not diagnosed in the United States


http://bse-atypical.blogspot.com/2009/05/identification-and-characterization-of.html



Wednesday, February 11, 2009

Atypical BSE North America Update

February 2009

Both of the BSE cases ascertained in the US native-born cattle were atypical cases (H-type), which contributed to the initial ambiguity of the diagnosis. 174, 185 In Canada, there have been 2 atypical BSE cases in addition to the 14 cases of the classic UK strain of BSE2: one was the H-type, and the other was of the L-type.198 snip...end source : Enhanced Abstract Journal of the American Veterinary Medical Association January 1, 2009, Vol. 234, No. 1, Pages 59-72 Bovine spongiform encephalopathy Jane L. Harman, DVM, PhD; Christopher J. Silva, PhD


http://avmajournals.avma.org/doi/ref/10.2460/javma.234.1.59



Atypical BSE North America Update February 2009


http://bse-atypical.blogspot.com/2009/02/atypical-bse-north-america-update.html



Thursday, April 30, 2009 FDA Issues Final Guidance for Renderers on Substances Prohibited From Use in Animal Food or Feed CVM Update Back April 30, 2009


http://madcowfeed.blogspot.com/2009/04/fda-issues-final-guidance-for-renderers.html



Sunday, May 10, 2009

Meeting of the Transmissible Spongiform Encephalopathies Committee On June 12, 2009 (Singeltary submission)


http://tseac.blogspot.com/2009/05/meeting-of-transmissible-spongiform.html



April 20, 2009

National Prion Disease Pathology Surveillance Center Cases Examined1 (December 31, 2008)

National Prion Disease Pathology Surveillance Center Cases Examined1 (December 31, 2008)

Year Total Referrals2 Prion Disease Sporadic Familial Iatrogenic vCJD 1996 & earlier 42 32 28 4 0 0 1997 115 68 59 9 0 0 1998 93 53 45 7 1 0 1999 115 69 61 8 0 0 2000 151 103 89 14 0 0 2001 210 118 108 9 0 0 2002 258 147 123 22 2 0 2003 273 176 135 41 0 0 2004 335 184 162 21 0 13 2005 346 193 154 38 1 0 2006 380 192 159 32 0 14 2007 370 212 185 26 0 0 2008 383 228 182 23 0 0 TOTAL 30715 17756 1490 254 4 2 1 Listed based on the year of death or, if not available, on year of referral; 2 Cases with suspected prion disease for which brain tissue and/or blood (in familial cases) were submitted; 3 Disease acquired in the United Kingdom; 4 Disease acquired in Saudi Arabia; 5 Includes 20 cases in which the diagnosis is pending, and 17 inconclusive cases; 6 Includes 25 cases with type determination pending in which the diagnosis of vCJD has been excluded. Rev 2/13/09 National


http://www.cjdsurveillance.com/pdf/case-table.pdf



http://www.cjdsurveillance.com/resources-casereport.html



http://www.aan.com/news/?event=read&article_id=4397&page=72.45.45



*5 Includes 20 cases in which the diagnosis is pending, and 17 inconclusive cases; *6 Includes 25 cases with type determination pending in which the diagnosis of vCJD has been excluded.


Greetings,

it would be interesting to know what year these atypical cases occurred, as opposed to lumping them in with the totals only.

are they accumulating ?

did they occur in one year, two years, same state, same city ?

location would be very interesting ?

age group ?

sex ?

how was it determined that nvCJD was ruled out ?

from 1997, the year i started dealing with this nightmare, there were 28 cases (per this report), up until 2007 where the total was 185 cases (per this report), and to date 2008 is at 182. a staggering increase in my opinion, for something that just happens spontaneously as some would have us believe. i don't believe it, not in 85%+ of all sporadic CJD cases. actually, i do not believe yet that anyone has proven that any of the sporadic CJD cases have been proven to be a spontaneous misfolding of a protein. there are many potential routes and sources for the sporadic CJD's. ...TSS


Sunday, April 12, 2009

r-calf and the USA mad cow problem, don't look, don't find, and then blame Canada


http://prionunitusaupdate2008.blogspot.com/2009/04/r-calf-and-usa-mad-cow-problem-dont.html



snip...


SEE FULL TEXT BELOW !

Monday, April 20, 2009 National Prion Disease Pathology Surveillance Center Cases Examined1 (December 31, 2008)


http://prionunitusaupdate2008.blogspot.com/2009/04/national-prion-disease-pathology.html



Friday, November 30, 2007

CJD QUESTIONNAIRE USA CWRU AND CJD FOUNDATION

snip...

i am reminded of a few things deep throat told me years ago;


=================================================2001


The most frightening thing I have read all day is the report of Gambetti's finding of a new strain of sporadic cjd in young people......... Dear God, what in the name of all that is holy is that!!! If the US has different strains of scrapie..... why???? than the UK... then would the same mechanisms that make different strains of scrapie here make different strains of BSE... if the patterns are different in sheep and mice for scrapie..... could not the BSE be different in the cattle, in the mink, in the humans....... I really think the slides or tissues and everything from these young people with the new strain of sporadic cjd should be put up to be analyzed by many, many experts in cjd........ bse..... scrapie


Scrape the damn slide and put it into mice..... wait..... chop up the mouse brain and and spinal cord........ put into some more mice..... dammit amplify the thing and start the damned research..... This is NOT rocket science... we need to use what we know and get off our butts and move.... the whining about how long everything takes..... well it takes a whole lot longer if you whine for a year and then start the research!!!



Not sure where I read this but it was a recent press release or something like that: I thought I would fall out of my chair when I read about how there was no worry about infectivity from a histopath slide or tissues because they are preserved in formic acid, or formalin or formaldehyde..... for God's sake........ Ask any pathologist in the UK what the brain tissues in the formalin looks like after a year....... it is a big fat sponge... the agent continues to eat the brain ...... you can't make slides anymore because the agent has never stopped........ and the old slides that are stained with Hemolysin and Eosin...... they get holier and holier and degenerate and continue... what you looked at 6 months ago is not there........ Gambetti better be photographing every damned thing he is looking at.....



Okay, you need to know. You don't need to pass it on as nothing will come of it and there is not a damned thing anyone can do about it. Don't even hint at it as it will be denied and laughed at.......... USDA is gonna do as little as possible until there is actually a human case in the USA of the nvcjd........ if you want to move this thing along and shake the earth.... then we gotta get the victims families to make sure whoever is doing the autopsy is credible, trustworthy, and a saint with the courage of Joan of Arc........ I am not kidding!!!! so, unless we get a human death from EXACTLY the same form with EXACTLY the same histopath lesions as seen in the UK nvcjd........ forget any action........ it is ALL gonna be sporadic!!! And, if there is a case....... there is gonna be every effort to link it to international travel, international food, etc. etc. etc. etc. etc. They will go so far as to find out if a sex partner had ever traveled to the UK/europe, etc. etc. .... It is gonna be a long, lonely, dangerous twisted journey to the truth. They have all the cards, all the money, and are willing to threaten and carry out those threats.... and this may be their biggest downfall...



Thanks as always for your help. (Recently had a very startling revelation from a rather senior person in government here.......... knocked me out of my chair........ you must keep pushing. If I was a power person.... I would be demanding that there be at least a million bovine tested as soon as possible and agressively seeking this disease. The big players are coming out of the wood work as there is money to be made!!!



In short: "FIRE AT WILL"!!! for the very dumb.... who's "will"! "Will be the burden to bare if there is any coverup!"



again it was said years ago and it should be taken seriously.... BSE will NEVER be found in the US!



As for the BSE conference call... I think you did agreat service to freedom of information and making some people feign integrity... I find it scary to see that most of the "experts" are employed by the federal government or are supported on the "teat" of federal funds. A scary picture! I hope there is a confidential panel organized by the new government to really investigate this thing.



You need to watch your back........ but keep picking at them....... like a buzzard to the bone... you just may get to the truth!!! (You probably have more support than you know. Too many people are afraid to show you or let anyone else know. I have heard a few things myself... you ask the questions that everyone else is too afraid to ask.)



================================================



greetings again voice,

then i remind everyone to read this;

'As implied in the Inset 25 we must not assume that transmission of BSE to other species will invariably present pathology typical of a scrapie-like disease.'



http://www.bseinquiry.gov.uk/files/yb/1991/01/04004001.pdf



Saturday, May 2, 2009

U.S. GOVERNMENT SUES WESTLAND/HALLMARK MEAT OVER USDA CERTIFIED DEADSTOCK DOWNER COW SCHOOL LUNCH PROGRAM



http://downercattle.blogspot.com/2009/05/us-government-sues-westlandhallmark.html



Sunday, May 17, 2009

WHO WILL WATCH THE CHILDREN ? SCHOOL LUNCH PROGRAM FROM DOWNER CATTLE UPDATE



http://downercattle.blogspot.com/2009/05/who-will-watch-children.html



CWRU CJD QUESTIONNAIRE HISTORY


http://cjdquestionnaire.blogspot.com/



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