Wednesday, November 28, 2012

Scientific and technical assistance on the provisional results of the study on genetic resistance to Classical scrapie in goats in Cyprus 1

SCIENTIFIC REPORT OF EFSA


Scientific and technical assistance on the provisional results of the study on genetic resistance to Classical scrapie in goats in Cyprus1


European Food Safety Authority2, 3


European Food Safety Authority (EFSA), Parma, Italy


ABSTRACT


This EFSA Scientific Report reviews and discusses the provisional results of a study (EURL/Cypriot study) on genetic resistance to Classical scrapie in goats in Cyprus. It is concluded that the provisional results obtained in the study further support the lower susceptibility to Classical scrapie in goats carrying the D146 and S146 alleles compared to wild type (N146N) goats. The results from intracerebral challenge are not compatible with a level of resistance as high as the one observed in sheep carrying the ARR allele or in goats carrying the K222 allele. Final results from the oral challenge will be crucial in determining the level of resistance associated with the D146 and S146 alleles. Furthermore, it is concluded that the provisional results obtained in the study are compatible with the possibility to use the D146 and S146 alleles to build a genetic strategy to control and eradicate Classical scrapie in goats in Cyprus. However, the success of such a strategy will be determined by the level of resistance associated with the D146 and S146 alleles against infection with all the different TSE agents proved to be circulating in Cyprus, which at this stage of the EURL/Cypriot study remains to be definitively assessed. In addition, as compared to the results of the model developed in the study, it is concluded that the efficiency of the implementation in the field of a breeding strategy selecting for the D146 and S146 alleles may be lower due to potential practical constraints related to the management of genetic diversity, to the selection for production and health traits and to the need of moving animals for breeding purposes in Cyprus. Recommendations on aspects that may be considered when completing the study are formulated.


© European Food Safety Authority, 2012


KEY WORDS


TSE, Classical scrapie, goat, genetic resistance, breeding programme, Cyprus.


snip...


Interim Conclusions


The oral challenge studies, the management cull and the whole herd cull data all support the previously published field observations that on one side the genotype N146N is particularly associated with scrapie susceptibility and on the other side the genotypes N146D, N146S, D146D, D146S and S146S are associated with a degree of resistance.


All genotypes can succumb to challenge by the intracerebral route, but the resulting phenotype is different when comparing the animals carrying genotype N146N with all the others (most notably, no detection of PrPSc in the periphery of infected animals with all the non-N146N-genotypes).


It is important that the oral challenges are continued to endpoint to establish the relative resistance of other genotypes to challenge by this more natural route Appendix A to Scientific Report of EFSA, EFSA Journal 2012;10(11):2972


Cyprus/EURL resistance in goats protocol Report May 2012


Page 13 of 13


All components of the study reinforce previously published UK caprine data3 which indicates that the current ELISA rapid test screen has considerably lower sensitivity (approx 50%) than immunohistochemistry.


The wider issue of discriminatory testing for BSE vs scrapie may need to be reviewed (regardless of genotype), since data from these studies suggest that direct extrapolation from ovine data may not be appropriate for all caprine isolates.


The TSE European Union Reference Laboratory at the Animal Health and Veterinary Laboratory Agency New Haw, UK


The Government Veterinary Services, Cyprus


May 2012







Wednesday, January 18, 2012


BSE IN GOATS CAN BE MISTAKEN FOR SCRAPIE


February 1, 2012



posted January 18, 2012



BSE in goats can be mistaken for scrapie



Bovine spongiform encephalopathy in goats could be misdiagnosed as scrapie in the absence of appropriate discriminatory tests, and such misidentification occurred at least once before such tests were developed, according to a report released in December.



The article, "Isolation of prion with BSE properties from farmed goat" (Emerging Infectious Diseases 2011;17:2253-2261), indicates BSE can affect small ruminants under natural conditions and that the condition can be misdiagnosed. The agent that causes scrapie is not known to infect humans, but consumption of beef contaminated with the prions that cause BSE is connected with variant Creutzfeldt-Jakob disease, a neurodegenerative disorder in humans.



The report calls for continued extensive surveillance and breeding plans to prevent BSE outbreaks among small ruminants. Such outbreaks could harm public health.



The authors stated in the text that the misdiagnosis occurred in 1990 in the United Kingdom. The case had been identified as suspected BSE in 2006 because differential immunohistochemical analysis of fixed brain tissue produced a signature indistinguishable from BSE. The authors of the recent report used a bioassay to confirm the BSE diagnosis.



The sample collected in 1990 was among 26 historic samples collected from 1984-2002, the report states.



The report indicates the U.K. goat and a goat in France found to have BSE in 2005 both likely became infected through contaminated food supplements.



While BSE lesions are contained mainly within nervous tissue in cattle, the report states "in small ruminants the BSE agent is widely distributed in peripheral tissues and can be transmitted horizontally." Feed ban measures alone would be insufficient for controlling a BSE outbreak in small ruminants, according to the report.



"Also, it would be impossible to prevent BSE from entering the human food chain through consumption of food products derived from small ruminants," the report states.













Discussion




We confirmed that the agent responsible for TSE in a UK goat, which was initially reported as scrapie in 1990 and subsequently as suspected BSE in 2006 (16), was a BSE agent. This conclusion was based on bioassay of nervous tissue in mice demonstrating similarities of histopathologic lesions, PrPSc mapping in the brain, and WB of PrPSc with those of mice inoculated with BSE from various ovine, caprine, and bovine sources.











Saturday, December 3, 2011




Isolation of Prion with BSE Properties from Farmed Goat Volume 17, Number




12—December 2011









snip...see full text ;








Scientific Opinion on genetic TSE resistance in goats in all European Union Member States Question number: EFSA-Q-2009-00448


Adopted: 21 October 2009 Summary (0.1Mb)


Opinion (0.3Mb)


Summary


Following a request from the European Commission, the Panel on Biological Hazards (BIOHAZ) was asked to deliver a scientific opinion on genetic resistance to Transmissible Spongiform Encephalopathies (TSE) in goats.


For a first part of that request, the BIOHAZ Panel adopted on 5th March 2009 a scientific opinion on the scientific validity of a study carried out by the Cypriot authorities under the auspices of the Community Reference Laboratory (CRL) for TSEs. That scientific opinion also indicated to what extent the information contained in the study could be used as relevant tools to control Classical scrapie in Cyprus.


In the current scientific opinion the BIOHAZ Panel addresses genetic resistance as a relevant tool for breeding for resistance to all TSEs of goats (including Atypical scrapie and BSE) in all the Member States (MSs) (except for Classical scrapie in Cyprus).


To carry out this task, available scientific knowledge on genetic TSE resistance in goats in the EU is reviewed, addressing those PRNP polymorphisms for which a capacity to provide resistance to TSEs in goats has been (or is being) investigated. Details tailored to the different TSEs found in this small ruminants (i.e. Classical scrapie, Atypical scrapie and BSE) are also considered and presented.


Further on, the feasibility of a large-scale breeding program in animal populations would need to be supported by a sound logistical and technical infrastructure in any given territory. In order to collect preliminary data that could help to evaluate the specific situation in the different EU MSs, a questionnaire was developed and circulated among the EFSA BSE-TSE Network. The results of the analysis of the replies received are also presented herewith.


The BIOHAZ Panel concluded that there are encouraging but as yet incomplete data to consider supporting a breeding programme for resistance in goats against Classical scrapie in all EU MSs, and ongoing studies are expected to provide a more robust scientific background in the coming years. On the other hand, at this moment there are not enough data available to consider supporting a breeding programme for resistance against Atypical scrapie and BSE in goats in all EU MSs. Experiments are ongoing on BSE in goats and results will be available in the next years. Furthermore, there are limited data suggesting that an allele (H154) might confer resistance to Classical scrapie but increase susceptibility to Atypical scrapie.


The frequency of the wild type allele, which is known to confer susceptibility to Classical scrapie, is high in all goat breeds considered. Thus, selection for putative resistance alleles will be slow, complicated and highly dependent on breeding structure.


It is acknowledged that any large scale breeding programme for TSE resistance in goats must take into consideration key elements related to the current dissemination of potentially TSE protective polymorphisms in the goat population of each EU MS and the characterisation of the real protection provided by those polymorphisms. At present, only a few EU MSs seem to have in place the necessary elements to introduce a breeding for resistance programme for Classical scrapie in goats.


The BIOHAZ Panel makes a series of recommendations on new investigations in order to assess the efficacy of breeding for the candidate PRNP alleles as a mean to control TSEs in goats. Furthermore, research on the possible adverse effects of the candidate PRNP polymorphisms on other production traits should be encouraged. In addition, it is recommended that a breeding for resistance programme for TSE in goats is first implemented in the seven EU MSs with the largest goat population as this would have the most impact.


Published: 9 November 2009







OPINION








Tuesday, November 10, 2009


A retrospective immunohistochemical study reveals atypical scrapie has existed in the United Kingdom since at least 1987


Brief Research Reports








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


Subject: Twelve Greek goats were found to be suffering from the brain-wasting disease scrapie in the first half of 2004


Date: Wed, 19 Jan 2005 13:30:26 –0600


From: "Terry S. Singeltary Sr."


To: Bovine Spongiform Encephalopathy


CC: cjdvoice@yahoogroups.com


Wednesday January 19, 2005


Brain disease in Greek goats


Twelve Greek goats were found to be suffering from the brain-wasting disease scrapie in the first half of 2004, EU figures made public yesterday reveal.


The data, issued by the European Food Safety Authority (EFSA), show that 12 cases of transmissible spongiform encephalopathies (TSE) were discovered in Greece, eight in Cyprus and 26 in France out of some 17,294 goats tested throughout the EU in 2004. The figures were made public by Left Coalition Synaspismos MEP Dimitris Papadopoulos.


Some 100 Europeans have died from the human form of bovine spongiform encephalopathy (BSE), or mad cow disease, a strain of the TSE group. Meanwhile, tests are continuing in the case of a French goat slaughtered in 2002, which experts think may have developed BSE. The EU bans the use of milk and meat from herds affected by a TSE case.









TSS





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


Subject: Twelve Greek goats were found to be suffering from the brain-wasting disease scrapie in the first half of 2004


Date: Wed, 19 Jan 2005 13:30:26 –0600


From: "Terry S. Singeltary Sr."


To: Bovine Spongiform Encephalopathy


CC: cjdvoice@yahoogroups.com


Wednesday January 19, 2005


Brain disease in Greek goats


Twelve Greek goats were found to be suffering from the brain-wasting disease scrapie in the first half of 2004, EU figures made public yesterday reveal.


The data, issued by the European Food Safety Authority (EFSA), show that 12 cases of transmissible spongiform encephalopathies (TSE) were discovered in Greece, eight in Cyprus and 26 in France out of some 17,294 goats tested throughout the EU in 2004. The figures were made public by Left Coalition Synaspismos MEP Dimitris Papadopoulos.


Some 100 Europeans have died from the human form of bovine spongiform encephalopathy (BSE), or mad cow disease, a strain of the TSE group. Meanwhile, tests are continuing in the case of a French goat slaughtered in 2002, which experts think may have developed BSE. The EU bans the use of milk and meat from herds affected by a TSE case.










TSS





Comment from Terry S Singeltary, CJD WATCH/VOICE




Document ID: APHIS-2007-0033-0002 Document Type: Public Submission
This is comment on Proposed Rule: Agricultural Bioterrorism Protection Act of 2002; Biennial Review and Republication of the Select Agent and Toxin List
Docket ID:
RIN:0579-AC53

Topics: No Topics associated with this document

View Document:





snip...



Under APHIS-PPQ’s agriculture quarantine inspection monitoring, 584 air passengers from Greece were sampled for items of agricultural interest in fiscal year 2000. Of these passengers, 14 carried meat (non-pork) items that could potentially transmit pathogens that cause BSE; most passengers carried from one to two kilograms (kg) of meat, although one passenger in November 1999 carried 23 kg of meat in a suitcase. Florida, Massachusetts, and New York were the reported destinations of these passengers. None of the passengers with meat items reported plans to visit or work on a ranch or farm while in the US.





Source: US Department of Transportation, and APHIS-PPQ Agricultural Quarantine Inspection data base

















Monday, November 19, 2012

Prion in Saliva of Bovine Spongiform Encephalopathy–Infected Cattle

Letter



Prion in Saliva of Bovine Spongiform Encephalopathy–Infected Cattle



To the Editor: A definitive diagnosis of bovine spongiform encephalopathy (BSE) in cattle usually relies on Western blot and immunohistochemical testing of samples from the obex region of the brainstem. These conventional diagnostic tests can detect the presence of the abnormal (disease-associated) form of the prion protein (PrPSc) in brain samples several months before the onset of clinical signs; however, there is no appropriate, universal tool for early preclinical and antemortem diagnosis of BSE. Furthermore, confirmation of the disease is currently only possible by postmortem examination of brain tissues. In this study, we used the serial protein misfolding cyclic amplification (sPMCA) technique to determine the presence of PrPSc in saliva samples collected from BSE-infected cows before and after the onset of disease (1). In a previous study (2), we analyzed the tissue distribution of PrPSc in cattle up to 66 months after they were orally inoculated with a relatively low dose (5 g) of homogenized brainstem from animals with naturally occurring BSE in England. In 2011, after publication of that study and 83.3 months after the cows were inoculated, clinical signs of BSE developed in 1 cow (no. 5444); necropsy was performed 84.7 months after inoculation. In addition, we used saliva samples from 2 BSE-affected cows (nos. 5413 and 5437) (2) to determine the presence of PrPSc. We collected saliva samples from animals at 4 monthly intervals, beginning in 2009, 56 months after inoculation. Samples were stored at −80°C until analysis. Using the sodium phosphotungstic acid precipitation method, we concentrated (100-fold) individual 1-mL saliva samples from each time point. We then diluted the concentrated samples 1:10 with the normal isoform of prion protein substrate containing 0.5% potassium dextran sulfate. Using the sPMCA technique as described (1), we amplified the samples in 3–8 tubes, and we used Western blot to analyze the proteinase K–treated sPMCA products (2). Using Western blot and immunohistochemical tests, we detected the accumulation of PrPSc in brains collected at necropsy from the 3 cows examined. In addition, using the sPMCA technique, we detected PrPSc signal in 1) saliva samples that were concentrated from samples collected from the same 3 cows at necropsy and in 2) concentrated saliva samples that were collected from 2 of the cows (nos. 5413 and 5444) at the early clinical stages of disease.




Figure



Figure. . Western blot detection, using the serial protein misfolding cyclic amplification technique, of the abnormal (disease-associated) form of the prion protein (PrPSc) in concentrated saliva samples from 3 cows experimentally infected...



After saliva samples underwent 3 rounds of amplification, we detected PrPSc in a saliva sample that was collected from cow number 5437 two months before the clinical onset of clinical symptoms (Figure). For 2 of the cows (nos. 5413 and 5437), the positive ratio of salivary PrPSc at round 4 of amplification increased as the disease progressed (Figure). Because PrPSc signal could be detected in BSE-infected brain homogenates diluted up to 10−10 after 2 rounds of amplification (1), we estimated PrPSc levels in the nonconcentrated original saliva samples to be lower than those in BSE-infected brain homogenate diluted to 10−12. No PrPSc signal was detected in samples collected from the 3 cows 3–5 months before the onset of clinical symptoms or from age-matched noninfected controls, even after 4 rounds of amplification. We demonstrated the presence of PrPSc in saliva of BSE-affected cows during the clinical stage of the disease, and in 1 case, at the preclinical or asymptomatic stage. Our findings suggest that PrPSc is likely to be detected in the saliva of BSE-affected cattle during the clinical stage of disease, after accumulation of PrPSc in the brain. PrPSc was found in the salivary glands of BSE-affected cattle at the terminal stage of infection (1). Therefore, once the infectious agent reaches the central nervous system, it may spread centrifugally from the brain to the salivary glands through the autonomic nervous system. Infectivity of saliva and the presence of PrPSc in saliva have been reported in other ruminants affected with transmissible spongiform encephalopathy. Infectivity of saliva was demonstrated in deer with chronic wasting disease (3) and in scrapie-affected sheep (4); the immunolabeled PrPSc accumulated in the salivary glands of scrapie-affected sheep (5). A low level of PrPSc was detected in concentrated buccal swab samples of preclinical scrapie-infected sheep by using sPMCA (6,7). These results suggest that small amounts of PrPSc may accumulate in the salivary glands and are then secreted into saliva. The presence of infectious prions in saliva may explain the facile horizontal transmission of scrapie in sheep (4–6) and chronic wasting disease in deer (4,8). There has been no epidemiologic evidence, however, that saliva, milk, blood, and cerebrospinal fluid from BSE-infected cattle are infectious (9). Nonetheless, the potential risk for BSE transmission by body fluids or excretions from BSE-infected cattle is cannot be ruled out by the current data.



Hiroyuki Okada, Yuichi Murayama , Noriko Shimozaki, Miyako Yoshioka, Kentaro Masujin, Morikazu Imamura, Yoshifumi Iwamaru, Yuichi Matsuura, Kohtaro Miyazawa, Shigeo Fukuda, Takashi Yokoyama, and Shirou Mohri



Author affiliations: Author affiliations: National Agriculture and Food Research Organization, Tsukuba, Japan (H. Okada, Y. Murayama, N. Shimozaki, M. Yoshioka, K. Masujin, M. Imamura, Y. Iwamaru, Y. Matsuura, K. Miyazawa, T. Yokoyama, S. Mohri); Hokkaido Research Organization, Shintoku, Japan (S. Fukuda)



Acknowledgment



This work was supported by a grant-in-aid from the BSE and Other Prion Disease Project of the Ministry of Agriculture, Forestry and Fisheries, Japan.



References



1.Murayama Y, Yoshioka M, Masujin K, Okada H, Iwamaru Y, Imamura M, Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrPSc and enable ultrasensitive detection of bovine PrPSc. PLoS ONE. 2010;5:e13152. DOIPubMed 2.Okada H, Iwamaru Y, Imamura M, Masujin K, Matsuura Y, Murayama Y, Detection of disease-associated prion protein in the posterior portion of the small intestine involving the continuous Peyer’s patch in cattle orally infected with bovine spongiform encephalopathy agent. Transbound Emerg Dis. 2011;58:333–43. DOIPubMed 3.Haley NJ, Seelig DM, Zabel MD, Telling GC, Hoover EA. Detection of CWD prions in urine and saliva of deer by transgenic mouse bioassay. PLoS ONE. 2009;4:e4848. DOIPubMed 4.Tamgüney G, Richt JA, Hamir AN, Greenlee JJ, Miller MW, Wolfe LL, Salivary prions in sheep and deer. Prion. 2012;6:52–61. DOIPubMed 5.Vascellari M, Nonno R, Mutinelli F, Bigolaro M, Di Bari MA, Melchiotti E, PrPSc in salivary glands of scrapie-affected sheep. J Virol. 2007;81:4872–6. DOIPubMed 6.Maddison BC, Rees HC, Baker CA, Taema M, Bellworthy SJ, Thorne L, Prions are secreted into the oral cavity in sheep with preclinical scrapie. J Infect Dis. 2010;201:1672–6. DOIPubMed 7.Gough KC, Baker CA, Rees HC, Terry LA, Spiropoulos J, Thorne L, The oral secretion of infectious scrapie prions occurs in preclinical sheep with a range of PRNP genotypes. J Virol. 2012;86:566–71. DOIPubMed 8.Mathiason CK, Powers JG, Dahmes SJ, Osborn DA, Miller KV, Warren RJ, Infectious prions in the saliva and blood of deer with chronic wasting disease. Science. 2006;314:133–6. DOIPubMed 9.Brown P, Andréoletti O, Bradley R, Budka H, Deslys JP, Groschup M, WHO tables on tissue infectivity distribution in transmissible spongiform encephalopathies. Geneva: World Health Organization; 2010 [cited 2011 Nov 2]. http://www.who.int/bloodproducts/tablestissueinfectivity.pdf



Figure Figure. . Western blot detection, using the serial protein misfolding cyclic amplification technique, of the abnormal (disease-associated) form of the prion protein (PrPSc) in concentrated saliva samples from 3 cows experimentally...



Suggested citation for this article: Okada H, Murayama Y, Shimozaki N, Yoshioka M, Masujin K, Imamura M, et al. Prion in saliva of bovine spongiform encephalopathy–infected cattle [letter]. Emerg Infect Dis [Internet]. 2012 Dec [date cited]. DOI: http://dx.doi.org/10.3201/eid1812.120528



DOI: 10.3201/eid1812.120528









DISSERTATION



CHRONIC WASTING DISEASE: A MODEL FOR PRION TRANSMISSION VIA SALIVA AND URINE


Submitted by Nicholas James Haley


Department of Microbiology, Immunology and Pathology In partial fulfillment of the requirements For the Degree of Doctor of Philosophy Colorado State University Fort Collins, Colorado Summer 2010



ABSTRACT OF DISSERTATION CHRONIC WASTING DISEASE: A MODEL FOR PRION TRANSMISSION VIA SALIVA AND URINE



Chronic wasting disease (CWD) of cervids is a prion disease distinguished by its high level of transmissibility, wherein bodily fluids and excretions are thought to play an important role. Typical of all prion diseases, CWD is characterized by the forced conversion of the normal prion protein (PrPC) into a misfolded isoform (PrPCWD), which has been shown to accumulate primarily in tissues of the lymphoid and nervous systems, though has also been found in other peripheral tissues including elements of the cardiovascular, musculoskeletal, and urogenital systems. Despite strong evidence that natural infection is acquired from the environment, as well as saliva and blood, a more thorough evaluation of excreta, including saliva, urine, and feces, is essential for a comprehensive foundation for (1) understanding how environmental CWDcontamination occurs, (2) developing in vitro assays for the antemortem identification of CWD-infected cervids, and (3) demonstrating the pathogenesis of the disease in the natural host.


In this dissertation, two approaches are used to identify infectious CWD prions and PrPCWD in the bodily fluids and tissues of CWD-exposed white-tailed deer: a novel bioassay system using a transgenic mouse line expressing the cervid PrP protein (Tg[CerPrP] mice), and a recently developed prion amplification assay known as serial iv protein misfolding cyclic amplification (sPMCA). In conjunction with immunohistochemistry and western blotting, these two assays were used to definitively identify CWD prions in saliva and urine, in addition to elements of the lymphoreticular system, central and peripheral nervous systems, and urogenital and oropharyngeal tissues. In initial experiments, concentrated urine and saliva samples from terminal CWD+ white-tailed deer, suspected of harboring infectious CWD prions, was assessed by Tg[CerPrP] bioassay and sPMCA. Authentic prion infectivity was detected in urine and saliva using both detection systems in the case of urine, though only mouse bioassay successfully demonstrated CWD prions in saliva. The concentration of abnormal prion protein in bodily fluids was very low, as indicated by: undetectable PrPCWD levels by traditional assays (western blot, ELISA) and prolonged incubation periods and incomplete TSE attack rates in inoculated Tg[CerPrP] mice. These findings helped to extend the understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections.


Based on the identification of CWD prions in saliva (“prionsialia”) and urine (“prionuria”), I next sought to determine whether deer previously exposed orally to urine and feces from CWD+ sources, while conventional test-negative, may actually be harboring very low level CWD infection not evident in the 19 month observation period in initial cervid bioassay studies. A selection of tissues, including those of the lymphoreticular and both central and peripheral nervous systems were fully examined, initially using Tg[CerPrP] bioassay to demonstrate true infectivity, and secondarily with sPMCA. Positive controls consisted of issues from CWD+ deer exposed orally to saliva; negative control tissue sets were collected from deer exposed orally and intracranially to


v


CWD-negative brain. PrPCWD was detected in the tissues of orally 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. Serial PMCA analysis of individual tissues identified that obex alone was positive in urine/feces exposed deer. PrPCWD was amplified from both LRS and neural tissues of positive control deer but not from the same tissues of negative control deer. Detection 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.


Despite the confirmation of infectious prions in urine and saliva, along with conventional test-negative deer exposed to urine and feces, the manner in which infectivity is transferred to these excreta is unknown. To address this, I went on to apply sPMCA to tissues associated with production and excretion of urine and saliva in an effort to seek proximal sources of prion shedding. I blindly analyzed oropharyngeal and urogenital tissues, reproducibly demonstrating PrPCWD in each tissue examined in 3 rounds of sPMCA; whereas blood samples from the same animals and concurrent negative controls remained negative. Tissue distribution was affected by route of inoculation and CNS burden. The identification of PrPCWD in bodily fluids and conventional-test negative tissues – in the absence of detection by conventional methods – may indicate the presence of protease-sensitive infectious prions in excretory tissues not revealed by assays employing PK digestion or other means to remove PrPC reactivity.


vi


The continued evaluation of bodily fluids and peripheral tissues via sPMCA may therefore allow additional insights into prion transmission, trafficking, and pathogenesis.


Nicholas James Haley Department of Microbiology, Immunology and Pathology Colorado State University Fort Collins, CO 80523 Summer 2010


snip...


In summary, this study demonstrates for the first time amplifiable PrPCWD in various organs and tissues associated with prionsialia and prionuria. The ultimate source and mechanism of release into bodily fluids remain unknown, though elevated levels in both salivary gland and urinary bladder provides strong evidence that these tissues play a crucial role in prion excretion. In addition, the source and route of inoculation weighed heavily on the terminal peripheral distribution of PrPCWD, as did an individual’s apparent central nervous system burden. Finally, while this discovery provides evidence for prion invasion of peripheral excretory tissues, the timing of infiltration during CWD infection and the protease resistance profile of these prions warrant future studies in serial pathogenesis and detection of alternate infectious prion species.












Sunday, July 03, 2011


Prion Disease Detection, PMCA Kinetics, and IgG in Urine from Naturally/Experimentally Infected Scrapie Sheep and Preclinical/Clinical CWD Deer






Thursday, June 09, 2011


Detection of CWD prions in salivary, urinary, and intestinal tissues of deer: potential mechanisms of prion shedding and transmission





CHRONIC WASTING DISEASE: A MODEL FOR PRION TRANSMISSION VIA SALIVA AND URINE






Sunday, December 06, 2009


Detection of Sub-Clinical CWD Infection in Conventional Test-Negative Deer Long after Oral Exposure to Urine and Feces from CWD+ Deer






Wednesday, March 18, 2009


Detection of CWD Prions in Urine and Saliva of Deer by Transgenic Mouse Bioassay






*** Tuesday, September 02, 2008


Detection of infectious prions in urine (Soto et al Available online 13 August 2008.)






Friday, October 26, 2012


CHRONIC WASTING DISEASE CWD PENNSYLVANIA GAME FARMS, URINE ATTRACTANT PRODUCTS, BAITING, AND MINERAL LICKS






TSS


Tuesday, November 6, 2012

Transmission of New Bovine Prion to Mice, Atypical Scrapie, BSE, and Sporadic CJD, November-December 2012 update

ATYPICAL BSE AND SPORADIC CJD NOVEMBER-DECEMBER 2012


Dispatch


Transmission of New Bovine Prion to Mice


Thierry G.M. Baron* , Anne-Gaëlle Biacabe*, Anna Bencsik*, and Jan P.M. Langeveld† Author affiliations: *Agence Française de Sécurité Sanitaire des Aliments, Lyon, France; †Central Institute for Animal Disease Control, Lelystad, the Netherlands Abstract


We previously reported that cattle were affected by a prion disorder that differed from bovine spongiform encephalopathy (BSE) by showing distinct molecular features of disease-associated protease-resistant prion protein (PrPres). We show that intracerebral injection of such isolates into C57BL/6 mice produces a disease with preservation of PrPres molecular features distinct from BSE.


Conclusions Our data show that the recently identified bovine H-type isolates involve an infectious agent that can induce development of a disease across a species barrier, while maintaining the specific associated PrPres molecular signature. This evidence in favor of a new bovine prion strain in cattle suggests that BSE is not the only transmissible prion disease in cattle. The origin of such cases has not been determined (7). These cases suggest either the existence of alternative origins of such diseases in cattle or phenotypic changes of PrPres after infection with the BSE agent. However, based on analysis of molecular features of prion diseases in cattle, this situation is similar to that in humans (5), in which different subtypes of sporadic Creutzfeldt-Jakob disease agents are found.


Dr Baron is head of the Unité Agents Transmissibles Non Conventionnels, Agence Française de Sécurité Sa


nitaire des Aliments, in Lyon. His research focuses on diagnosis of prion diseases of ruminants and characterization of the disease-associated prion protein and infectious agents, with particular emphasis on atypical forms of these diseases.




Dispatch


Differentiation of Prions from L-type BSE versus Sporadic Creutzfeldt-Jakob Disease


Simon Nicot, Anna Bencsik, Eric Morignat, Nadine Mestre-Francés, Armand Perret-Liaudet, and Thierry Baron


Author affiliations: Author affiliations: Agence Nationale de Sécurité Sanitaire (Anses), Lyon, France (S. Nicot, A. Bencsik, E. Morignat, T. Baron); INSERM U710, Montpellier, France; Université Montpellier 2, Montpellier; École Pratique des Hautes Études, Paris, France (N. Mestre-Francés); Hôpitaux Civils de Lyon, Université Lyon 1, INSERM U1028, and Centre National de la Recherche Scientifique, Lyon (A. Perret-Liaudet)


Abstract


We compared transmission characteristics for prions from L-type bovine spongiform encephalopathy and MM2-cortical sporadic Creutzfeldt-Jakob disease in the Syrian golden hamster and an ovine prion protein–transgenic mouse line and isolated distinct prion strains. Our findings suggest the absence of a causal relationship between these diseases, but further investigation is warranted.






Greetings,


THE abstract claims "absence of a causal relationship", is highly misleading and goes beyond what they actually studied which was quite limited to one case of mm scjd. a lot of passaging and cell lines can change properties. seems they are saying not all mm scjd comes from their one isolate of L bse, and if that is the case, they should have stated it that way. however, seems the author et al are grasping for straws to me, to make the c-BSE the only zoonotic TSE in the Bovine, where scientific studies on atypical BSE shows that indeed some atypical TSE are very similar to sporadic CJD. ...TSS


sporadic CJD and typical and atypical BSE and Scrapie


PO-030: Interspecies transmission of L-type bovine spongiform encephalopathy reveals similarities with transmissible mink encephalopathy but differences with sporadic (type MM2) Creutzfeldt-Jakob disease


Simon Nicot,1 Anna Bencsik,1 Nadine Mestre-Francés,2 Armand Perret-Liaudet,3 Thierry Baron1 1Agence Nationale de Sécurité Sanitaire (ANSES); Lyon, France; 2Institut National de la Santé et de la Recherche Médicale (IN SER M) U710; Montpellier, France; 3Hopitaux Civils de Lyon; Lyon, France


The L-type BSE (L-BSE) in cattle requires particular attention for public health in line with recent observations regarding its transmission in primates and human transgenic mice, in which it transmits more efficiently than the classical BSE. Moreover, previous reports suggested a possible relationship between L-BSE and some human subtypes of sporadic CJD (type 2).1,2


We conducted a comparative transmission study of L-BSE and a case of sporadic CJD (subtype MM2) using a combination of different rodent models. To allow a genetic proximity of these diseases, an isolate of L-BSE experimentally-passaged in a nonhuman primate3 was included for comparison. We previously described the transmission properties and phenotypes of L-BSE in different models, including Syrian hamsters as well as ovine and bovine transgenic mice.


Remarkably, these features were shown to be indistinguishable from those of the Stetsonville Transmissible Mink Encephalopathy (TME) agent in these models, supporting the view of a cross-species foodborne transmission of L-BSE as the origin of this TME outbreak in North America.


When challenged with human sporadic CJD (type MM2) prions, these models showed different susceptibilities, in contrast to results obtained with L-BSE, even after a first passage in a nonhuman primate. Among these rodent models, the ovine transgenic mouse line was remarkably susceptible to all prion sources, including not only L-BSE and TME, but also the sporadic CJD MM2 case, allowing the comparison of these diseases in this model.


Taken together, the transmission data obtained suggest that L-BSE and sporadic CJD MM2 have divergent biological properties, together with distinct histopathological and biochemical characteristics. These results do not support the hypothesis of a common origin of these diseases, which are likely to involve distinct agent strains.


These results encourage further investigation of this approach for a complete evaluation of the potential relations between L-BSE and human sporadic CJD, notably by using additional human cases of sporadic MM2 CJD, as well as other subtypes of sporadic CJD, considering the prion diversity existing in humans.


References


1. Casalone C, Zanusso G, Acutis P, Ferrari S, Capucci L, Tagliavini F, et al. Identification of a second bovine amyloidotic spongiform encephalopathy: molecular similarities with sporadic Creutzfeldt-Jakob disease. Proc Natl Acad Sci U S A 2004; 101:3065-70; PMID:14970340; http://dx.doi.org/10.1073/pnas.0305777101.


2. Comoy EE, Casalone C, Lescoutra-Etchegaray N, Zanusso G, Freire S, Marcé D, et al. Atypical BSE (BASE) transmitted from asymptomatic aging cattle to a primate. PLoS One 2008; 3:e3017; PMID:18714385; http://dx.doi.org/10.1371/journal. pone.0003017.


3. Mestre-Francés N, Nicot S, Rouland S, Biacabe AG, Quadrio I, Perret-Liaudet A, et al. Oral transmission of L-type bovine spongiform encephalopathy in primate model. Emerg Infect Dis 2012; 18:142-5; PMID:22261009; http://dx.doi.org/10.3201/ eid1801.111092.




PO-028: Oral transmission of L-type bovine spongiform encephalopathy (L-BSE) in primate model Microcebus murinus


Nadine Mestre-Frances,1 Simon Nicot,2 Sylvie Rouland,1 Anne-Gaëlle Biacabe,2 Isabelle Quadrio,3 Armand Perret-Liaudet,3 Thierry Baron,2 Jean-Michel Verdier1 1IN SER M UM2; Montpellier, France; 2Anses; Lyon, France; 3Hopitaux Civils de Lyon; Lyon, France


An atypical form of bovine spongiform encephalopathy has been identified in cattle in Europe, North America and Japan and was designed as L-type BSE (L-BSE) due to the lower apparent molecular mass of the unglycosylated, protease-resistant prion protein (PrPres) detected by western blot compared with classical BSE. Experimental evidences from studies in transgenic mice expressing human PrP and in primate models suggest a higher risk of transmission to humans of the L-BSE form than for classical BSE agent. However, a major unresolved issue concerns the potential transmissibility of the L-BSE agent by oral route. To address this question, we infected mouse lemurs (Microcebus murinus), a non-human primate model, with L-BSE by intracerebral or oral route.


Four adult lemurs were intracerebrally (IC) inoculated with 5mg of L-BSE infected brain homogenate of an atypical French BSE case (02-2528). Four young and four adult animals were fed with 5 mg or 50 mg of infected brain. After sacrifice, the brain tissues were biochemically and immunocytochemically investigated for PrPres.


The 4 animals IC inoculated died at 19 and 22 months postinoculation (mpi). They developed blindness, tremor, abnormal posture, incoordinated movements, balance loss. Symptoms get worse according to the disease progression, until severe ataxia. Severe spongiosis was evidenced into the thalamus, the striatum, the mesencephalon, and the brainstem, whereas into the cortex the vacuolisation was weaker. Strong deposits of PrPres were detected into the thalamus, the striatum, and the hippocampus whereas in the cerebral cortex, PrPres was prominently accumulated in plaques.


The orally inoculated animals showed similar clinical symptoms occurring between 27 and 34 mpi. Disease was characterized by progressive prostration, loss of appetite and poor appearance of the fur. Only one adult animal showed disequilibrium. PrPres was strongly accumulated only in the striatum and thalamus and weakly into the cortex. No plaques were evidenced. Two animals that were orally challenged at the age of two years are still alive and healthy 34 months after inoculation. The western blot analysis showed uniform molecular profiles, irrespective of the route or dose of infection, and included notably a PrPres form with low apparent molecular mass (~19 kDa) similar to the PrPres in the original cattle brain. However, the PrPres profile in lemurs was characterized by a higher proportion of di- and mono-glycosylated species (up to 95% of the total signal) than in the bovine L-BSE inoculum (~80%). In addition, small amounts of PrPres were detected by western blotting in the spleen of three animals (one intra-cerebrally inoculated and two fed with 5 mg of cattle brain).


Here, we demonstrate that the L-BSE agent can be transmitted by oral route from cattle to young and adult mouse lemurs. In comparison to IC inoculated animals, orally challenged lemurs were characterized by longer survival periods as expected with this route of infection. Our study now clearly confirms experimentally the potential risk of interspecies oral transmission of L-BSE. It is thus very important to maintain the precautionary measures that prevent the entry into the food chain of potentially infected tissues from cattle.




PO-044: MM2-thalamic Creutzfeldt-Jakob disease— Neuropathological, biochemical and transmission studies identify a distinctive prion strain


Silvia Suardi,1 Fabio Moda,1 Giuseppe Di Fede,1 Antonio Indaco,1 Margherita Ruggerone,1 Ilaria Campagnani,1 Jan Langeveld,2 Alessandro Terruzzi,3 Antonio Brambilla,4 Pietro Zerbi,5 Paolo Fociani,5 Matthew Bishop,6 Robert Will,6 Jean Manson,7 Giorgio Giaccone,1 Fabrizio Tagliavini1 1Neurological Institute C. Besta; Milan, Italy; 2Central Veterinary Institute; Lelystad, Netherlands; 3Policlinico S. Marco; Bergamo, Italy; 4Azienda Ospedaliera Bolognini; Seriate, Italy; 5Ospedale Luigi Sacco; Milan, Italy; 6National Creutzfeldt-Jakob Disease Surveillance Unit; Western General Hospital; Edinburgh, UK; 7The Roslin Institute and R(D)SVS; Edinburgh, UK


Introduction. In Creutzfeldt-Jakob disease (CJD), molecular typing based on the size of the protease resistant core of the disease- associated prion protein (PrPSc) and the M/V polymorphism at codon 129 of the PRNP gene correlates with the clinico-pathologic subtypes. Approximately 95% of the sporadic 129MM CJD patients are characterized by cerebral deposition of type 1 PrPSc and correspond to the classic clinical CJD phenotype. The rare 129MM CJD patients with type 2 PrPSc (MM2) are further subdivided in a cortical and a thalamic form, also indicated as sporadic Fatal Insomnia (sFI). To define the diversity between the MM2 subtypes, we characterized two different cases of MM2-thalamic CJD in terms of clinical, neuropathological, biochemical and transmission properties.


Material and Methods. Two young patients, both MM at codon 129 and affected by a thalamic form of CJD, were followed up to describe the clinical course and, after autopsy, the neuropathological and biochemical features were characterized. To evaluate the transmission properties, a group of gene targeted transgenic mice expressing human PrP on a mouse PrP knock-out background and carrying the MM 129 codon genotype (HuMM) were injected by a combination of intracerebral (i.c.) (20 μl) and intraperitoneal (i.p.) (100 μl) routes with 10% homogenate of cerebral cortex from one of the two MM2-thalamic cases.


Results. Main neuropathological features in the two cases were diffuse, synaptic PrP immunoreactivity in the cerebral cortex and severe neuronal loss and gliosis in the thalamus and olivary nucleus. Western blot analysis showed the presence of type 2A PrPSc. Challenge of transgenic mice expressing 129MM human PrP showed that MM2-thalamic sCJD transmitted the disease to 13 out of 14 mice, with an incubation time of 535 ± 32 (mean ± s.e.m) days and a survival period of 557 ± 23 (mean ± s.e.m) days. All affected mice showed mild spongiform changes in the brain and the presence of a type 2A PrPSc. Conversely, no clinical signs, neuropathological changes and PrPSc accumulation were observed in mice (n = 16) injected with a MM2-cortical case, up to 650 days post-inoculation.


Conclusions. Our data indicate that the MM2-thalamic form of CJD shows peculiar clinical, neuropathological and biochemical characteristics, and is capable to transmit the disease to mice expressing the human 129MM PRNP, at variance with MM2-cortical sCJD. The affected mice showed deposition of type 2A PrPSc, a scenario that is unprecedented in this mouse line. These data indicate that MM2-thalamic sCJD is caused by a prion strain distinct from the other sCJD subtypes including the MM2-cortical form.






Research


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


Thierry G.M. Baron* , Anna Bencsik*, Anne-Gaëlle Biacabe*, Eric Morignat*, and Richard A. Bessen† Author affiliations: *Agence Française de Sécurité Sanitaire des Aliments–Lyon, Lyon, France; †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).


Dr Baron is head of the Unit ATNC from Agence Française de Sécurité Sanitaire (AFSSA)–Lyon. His research focuses on prion diseases of ruminants, including diagnosis and characterization of the disease-associated prion protein and infectious agents, with particular emphasis on atypical forms of these diseases.






Evidence That Transmissible Mink Encephalopathy Results from Feeding Infected Cattle


Over the next 8-10 weeks, approximately 40% of all the adult mink on the farm died from TME.


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The rancher was a ''dead stock'' feeder using mostly (>95%) downer or dead dairy cattle...










Saturday, June 25, 2011


Transmissibility of BSE-L and Cattle-Adapted TME Prion Strain to Cynomolgus Macaque


Emmanuel Comoy,1,† Valérie Durand,1 Evelyne Correia,1 Sophie Freire,1 Jürgen Richt,2 Justin Greenlee,3 Juan-Maria Torres,4 Paul Brown,1 Bob Hills5 and Jean-Philippe Deslys1


1Atomic Energy Commission; Fontenay-aux-Roses, France; 2Kansas State University; Manhattan, KS USA; 3USDA; Ames, IA USA; 4INIA; Madrid, Spain; 5Health Canada; Ottawa, ON Canada†Presenting author; Email: emmanuel.comoy@cea.fr


The epidemiology of Transmissible mink encephalopathy (TME) indicates an alimentary origin. Several inter-species transmission experiments have not succeeded in establishing with certainty any natural reservoir of this prion strain, although both ovine and bovine sources have been suspected. Cattle exposed to TME develop a spongiform encephalopathy that is distinct from classical Bovine Spongiform Encephalopathy (c-BSE).


Inoculation of c-BSE to cynomolgus macaque provided early evidence of a possible risk to humans, and remains an important model to define the risk of both primary (oral transmission from cattle to primate) and secondary (intravenous intra-species transmission) exposures. We have also evaluated the transmissibility of other cattle prion strains to macaques, including L- and H- atypical forms of BSE, namely BSE-L and BSE-H, and cattle-adapted TME.


BSE-L induced a neurological disease distinct from c-BSE. Peripheral exposures demonstrate the transmissibility of BSE-L by oral, intravenous, and intra-cerebral routes, with incubation periods similar to c-BSE. Cattle-adapted TME also induced a rapid disease in cynomolgus macaque. The clinical features, lesion profile, and biochemical signature of the induced disease was similar to the features observed in animals exposed to BSE-L, suggesting a link between the two prion strains. Secondary transmissions to a common host (transgenic mouse overexpressing bovine PrP) of cattle-TME and BSE-L before or after passage in primates induced diseases with similar incubation periods: like the c-BSE strain, these cattle strains maintained their distinctive features regardless of the donor species and passages.


If the link between TME and BSE-L is confirmed, our results would suggest that BSE-L in North America may have existed for decades, and highlight a possible preferential transmission of animal prion strains to primates after passage in cattle.


=====================end...tss====================




link url not available, please see PRION 2011 ;






Monday, October 10, 2011


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


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EFSA and the European Centre for Disease Prevention and Control (ECDC) recently delivered a scientific opinion on any possible epidemiological or molecular association between TSEs in animals and humans (EFSA Panel on Biological Hazards (BIOHAZ) and ECDC, 2011). This opinion confirmed Classical BSE prions as the only TSE agents demonstrated to be zoonotic so far but the possibility that a small proportion of human cases so far classified as "sporadic" CJD are of zoonotic origin could not be excluded. Moreover, transmission experiments to non-human primates suggest that some TSE agents in addition to Classical BSE prions in cattle (namely L-type Atypical BSE, Classical BSE in sheep, transmissible mink encephalopathy (TME) and chronic wasting disease (CWD) agents) might have zoonotic potential.


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Thursday, August 12, 2010


Seven main threats for the future linked to prions


First threat


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


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


Second threat


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Rural and Regional Affairs and Transport References Committee


The possible impacts and consequences for public health, trade and agriculture of the Government's decision to relax import restrictions on beef Final report June 2010


2.65 At its hearing on 14 May 2010, the committee heard evidence from Dr Alan Fahey who has recently submitted a thesis on the clinical neuropsychiatric, epidemiological and diagnostic features of Creutzfeldt-Jakob disease.48 Dr Fahey told the committee of his concerns regarding the lengthy incubation period for transmissible spongiform encephalopathies, the inadequacy of current tests and the limited nature of our current understanding of this group of diseases.49


2.66 Dr Fahey also told the committee that in the last two years a link has been established between forms of atypical CJD and atypical BSE. Dr Fahey said that: They now believe that those atypical BSEs overseas are in fact causing sporadic Creutzfeldt-Jakob disease. They were not sure if it was due to mad sheep disease or a different form. If you look in the textbooks it looks like this is just arising by itself. But in my research I have a summary of a document which states that there has never been any proof that sporadic Creutzfeldt-Jakob disease has arisen de novo-has arisen of itself. There is no proof of that. The recent research is that in fact it is due to atypical forms of mad cow disease which have been found across Europe, have been found in America and have been found in Asia. These atypical forms of mad cow disease typically have even longer incubation periods than the classical mad cow disease.50




Atypical BSE in Cattle


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


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


This study will contribute to a correct definition of specified risk material (SRM) in atypical BSE. The incumbent of this position will develop new and transfer existing, ultra-sensitive methods for the detection of atypical BSE in tissue of experimentally infected cattle.






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.






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.


III International Symposium on THE NEW PRION BIOLOGY: BASIC SCIENCE, DIAGNOSIS AND THERAPY 2 - 4 APRIL 2009, VENEZIA (ITALY)




I ask Professor Kong ;


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


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


Professor Kong reply ;


.....snip


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


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


END...TSS


Thursday, December 04, 2008 2:37 PM


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


personal communication with Professor Kong. ...TSS


BSE-H is also transmissible in our humanized Tg mice.


The possibility of more than two atypical BSE strains will be discussed.


Supported by NINDS NS052319, NIA AG14359, and NIH AI 77774.












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: emmanuel.comoy@cea.fr




snip...




In summary, we have transmitted one atypical form of BSE (BASE) to a cynomolgus macaque monkey that had a shorter incubation period than monkeys infected with classical BSE, with distinctive clinical, neuropathological, and biochemical features; and have shown that the molecular biological signature resembled that seen in a comparatively uncommon subtype of sporadic CJD. We cannot yet say whether BASE is more pathogenic for primates (including humans) than cBSE, nor can we predict whether its molecular biological features represent a clue to one cause of apparently sporadic human CJD. However, the evidence presented here and by others justifies concern about a potential human health hazard from undetected atypical forms of BSE, and despite the waning epizoonosis of classical BSE, it would be premature to abandon the precautionary measures that have been so successful in reversing the impact of cBSE. We would instead urge a gradual, staged reduction that takes into account the evolving knowledge about atypical ruminant diseases, and both a permanent ban on the use of bovine central nervous system tissue for either animal or human use, and its destruction so as to eliminate any risk of environmental contamination.








Proc Natl Acad Sci U S A. 2004 March 2; 101(9): 3065–3070. Published online 2004 February 17. doi: 10.1073/pnas.0305777101 PMCID: PMC365745 Medical Sciences


Identification of a second bovine amyloidotic spongiform encephalopathy: Molecular similarities with sporadic Creutzfeldt-Jakob disease


Cristina Casalone,*† Gianluigi Zanusso,†‡ Pierluigi Acutis,* Sergio Ferrari,‡ Lorenzo Capucci,§ Fabrizio Tagliavini,¶ Salvatore Monaco,‡ and Maria Caramelli*


Abstract


Transmissible spongiform encephalopathies (TSEs), or prion diseases, are mammalian neurodegenerative disorders characterized by a posttranslational conversion and brain accumulation of an insoluble, protease-resistant isoform (PrPSc) of the host-encoded cellular prion protein (PrPC). Human and animal TSE agents exist as different phenotypes that can be biochemically differentiated on the basis of the molecular mass of the protease-resistant PrPSc fragments and the degree of glycosylation. Epidemiological, molecular, and transmission studies strongly suggest that the single strain of agent responsible for bovine spongiform encephalopathy (BSE) has infected humans, causing variant Creutzfeldt-Jakob disease. The unprecedented biological properties of the BSE agent, which circumvents the so-called ”species barrier” between cattle and humans and adapts to different mammalian species, has raised considerable concern for human health. To date, it is unknown whether more than one strain might be responsible for cattle TSE or whether the BSE agent undergoes phenotypic variation after natural transmission. Here we provide evidence of a second cattle TSE. The disorder was pathologically characterized by the presence of PrP-immunopositive amyloid plaques, as opposed to the lack of amyloid deposition in typical BSE cases, and by a different pattern of regional distribution and topology of brain PrPSc accumulation. In addition, Western blot analysis showed a PrPSc type with predominance of the low molecular mass glycoform and a protease-resistant fragment of lower molecular mass than BSE-PrPSc. Strikingly, the molecular signature of this previously undescribed bovine PrPSc was similar to that encountered in a distinct subtype of sporadic Creutzfeldt-Jakob disease.


snip...


Phenotypic Similarities Between BASE and sCJD. The transmissibility of CJD brains was initially demonstrated in primates (27), and classification of atypical cases as CJD was based on this property (28). To date, no systematic studies of strain typing in sCJD have been provided, and classification of different subtypes is based on clinical, neuropathological, and molecular features (the polymorphic PRNP codon 129 and the PrPSc glycotype) (8, 9, 15, 19). The importance of molecular PrPSc characterization in assessing the identity of TSE strains is underscored by several studies, showing that the stability of given disease-specific PrPSc types is maintained upon experimental propagation of sCJD, familial CJD, and vCJD isolates in transgenic PrP-humanized mice (8, 29). Similarly, biochemical properties of BSE- and vCJD-associated PrPSc molecules remain stable after passage to mice expressing bovine PrP (30). Recently, however, it has been reported that PrP-humanized mice inoculated with BSE tissues may also propagate a distinctive PrPSc type, with a ”monoglycosylated-dominant” pattern and electrophoretic mobility of the unglycosylated fragment slower than that of vCJD and BSE (31). Strikingly, this PrPSc type shares its molecular properties with the a PrPSc molecule found in classical sCJD. This observation is at variance with the PrPSc type found in M/V2 sCJD cases and in cattle BASE, showing a monoglycosylated-dominant pattern but faster electrophoretic mobility of the protease-resistant fragment as compared with BSE. In addition to molecular properties of PrPSc, BASE and M/V2 sCJD share a distinctive pattern of intracerebral PrP deposition, which occurs as plaque-like and amyloid-kuru plaques. Differences were, however, observed in the regional distribution of PrPSc. While in M/V2 sCJD cases the largest amounts of PrPSc were detected in the cerebellum, brainstem, and striatum, in cattle BASE these areas were less involved and the highest levels of PrPSc were recovered from the thalamus and olfactory regions.


In conclusion, decoding the biochemical PrPSc signature of individual human and animal TSE strains may allow the identification of potential risk factors for human disorders with unknown etiology, such as sCJD. However, although BASE and sCJD share several characteristics, caution is dictated in assessing a link between conditions affecting two different mammalian species, based on convergent biochemical properties of disease-associated PrPSc types. Strains of TSE agents may be better characterized upon passage to transgenic mice. In the interim until this is accomplished, our present findings suggest a strict epidemiological surveillance of cattle TSE and sCJD based on molecular criteria.




Thursday, June 21, 2012


Clinical and Pathologic Features of H-Type Bovine Spongiform Encephalopathy Associated with E211K Prion Protein Polymorphism


Atypical cases presumably of spontaneous or, in the case of E211K BSE-H, genetic origins highlight that it may not be possible to eradicate BSE entirely and that it would be hazardous to remove disease control measures such as prohibiting the feeding of meat and bone meal to ruminants.






bbbut ???




Thursday, June 21, 2012


Clinical and Pathologic Features of H-Type Bovine Spongiform Encephalopathy Associated with E211K Prion Protein Polymorphism






Friday, May 11, 2012


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




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




P03.141


Aspects of the Cerebellar Neuropathology in Nor98


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


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


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






PR-26


NOR98 SHOWS MOLECULAR FEATURES REMINISCENT OF GSS


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


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


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


119






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


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


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


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


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






Monday, December 1, 2008


When Atypical Scrapie cross species barriers


Authors


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


Content


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


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


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


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


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


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


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






Similarities between Forms of Sheep Scrapie and Creutzfeldt-Jakob Disease Are Encoded by Distinct Prion Types


Wiebke M. Wemheuer,* Sylvie L. Benestad,† Arne Wrede,* Ulf Schulze-Sturm,* Wilhelm E. Wemheuer,‡ Uwe Hahmann,* Joanna Gawinecka,§ Ekkehard Schu¨ tz,‡ Inga Zerr,§ Bertram Brenig,‡ Bjørn Bratberg,† Olivier Andre´ oletti,¶ and Walter J. Schulz-Schaeffer*


From the Prion and Dementia Research Unit,* Department of Neuropathology, and the National Transmissible Spongiform Encephalopathies Reference Center,§ Department of Neurology, University Medical Center Goettingen, Goettingen, Germany; the Department of Pathology,† National Veterinary Institute, Oslo, Norway; the Institute of Veterinary Medicine,‡ Faculty for Agricultural Sciences, University of Goettingen, Goettingen, Germany; and Animal Health,¶ Interactions Hôte Agent Pathogène, Ecole Nationale Ve´te´rinaire de Toulouse, Toulouse, France


Transmissible spongiform encephalopathies such as scrapie in sheep, Creutzfeldt-Jakob disease (CJD) in humans, and bovine sporadic encephalopathy in cattle are characterized by the accumulation of a misfolded protein: the pathological prion protein. Ever since bovine sporadic encephalopathy was discovered as the likely cause of the new variant of CJD in humans, parallels between human and animal transmissible spongiform encephalopathies must be viewed under the aspect of a disease risk for humans. In our study we have compared prion characteristics of different forms of sheep scrapie with those of different phenotypes of sporadic CJD. The disease characteristics of sporadic CJD depend considerably on the prion type 1 or 2. Our results show that there are obvious parallels between sporadic CJD type 1 and the so-called atypical/Nor98 scrapie. These parelleles apply to the deposition form of pathological prion protein in the brain, detected by the paraffin-embedded- tissue blot and the prion aggregate stability with regard to denaturation by the chaotropic salt guanidine hydrochloride. The same applies to sporadic CJD type 2 and classical scrapie. The observed parallels between types of sporadic CJD and types of sheep scrapie demonstrate that distinct groups of prion disease exist in different species. This should be taken into consideration when discussing interspecies transmission. (Am J Pathol 2009, 175:2566–2573; DOI: 10.2353/ajpath.2009.090623)


snip...


Discussion


In humans, different prion types are linked with clinically and neuropathologically distinct prion diseases.8 The present work emphasizes that the differences in deposition characteristics and stability with regard to denaturation between atypical/Nor98 and classical scrapie also account for different prion types. Moreover, the two scrapie types that have been characterized show a number of striking similarities with human PrPSc types in sporadic CJD. Hence, we propose that the existence of different PrPSc types might be a common denominator of prion diseases in humans and animals. Since these two prion types show an across-the-species comparability with similar biochemical and pathological characteristics, it is most likely that they exist due to a different conformational pattern of the disease-related prion protein.


snip...


Conclusion


As the prion protein is a highly conserved protein in terms of evolution, parallels between characteristics of prion types in TSEs of different species are of interest. In the present study, we report previously unknown similarities between sheep scrapie forms and human sporadic CJD types. We propose that the observed similarities between sheep scrapie and sporadic CJD in humans justify new interspecies groups of prion diseases in which prion types, not prion strains, are the major determinant for prion disease forms. While epidemiology implies that classical scrapie is not related to human TSEs,47 the atypical/Nor98 scrapie risk for human transmission has not yet been elucidated. Currently there is no compelling evidence that sCJD has a different origin than sporadic genesis. However, the finding of prion types with an across-the-species comparability might provide further understanding of the pathogenesis in prion diseases.


Prion Types Encode Interspecies TSEs 2571 AJP December 2009, Vol. 175, No. 6








Thursday, March 29, 2012


atypical Nor-98 Scrapie has spread from coast to coast in the USA 2012


NIAA Annual Conference April 11-14, 2011San Antonio, Texas








Wednesday, January 18, 2012


Selection of Distinct Strain Phenotypes in Mice Infected by Ovine Natural Scrapie Isolates Similar to CH1641 Experimental Scrapie


Journal of Neuropathology & Experimental Neurology: February 2012 - Volume 71 - Issue 2 - p 140–147






Thursday, July 14, 2011


Histopathological Studies of "CH1641-Like" Scrapie Sources Versus Classical Scrapie and BSE Transmitted to Ovine Transgenic Mice (TgOvPrP4)




Thursday, December 23, 2010


Molecular Typing of Protease-Resistant Prion Protein in Transmissible Spongiform Encephalopathies of Small Ruminants, France, 2002-2009


Volume 17, Number 1 January 2011






Thursday, November 18, 2010


Increased susceptibility of human-PrP transgenic mice to bovine spongiform encephalopathy following passage in sheep




Monday, October 10, 2011


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


snip...


EFSA and the European Centre for Disease Prevention and Control (ECDC) recently delivered a scientific opinion on any possible epidemiological or molecular association between TSEs in animals and humans (EFSA Panel on Biological Hazards (BIOHAZ) and ECDC, 2011). This opinion confirmed Classical BSE prions as the only TSE agents demonstrated to be zoonotic so far but the possibility that a small proportion of human cases so far classified as "sporadic" CJD are of zoonotic origin could not be excluded. Moreover, transmission experiments to non-human primates suggest that some TSE agents in addition to Classical BSE prions in cattle (namely L-type Atypical BSE, Classical BSE in sheep, transmissible mink encephalopathy (TME) and chronic wasting disease (CWD) agents) might have zoonotic potential.


snip...






see follow-up here about North America BSE Mad Cow TSE prion risk factors, and the ever emerging strains of Transmissible Spongiform Encephalopathy in many species here in the USA, including humans ;






Friday, February 11, 2011


Atypical/Nor98 Scrapie Infectivity in Sheep Peripheral Tissues




Sunday, December 12, 2010


EFSA reviews BSE/TSE infectivity in small ruminant tissues News Story 2 December 2010




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




5. A positive result from a chimpanzee challenged severly would likely create alarm in some circles even if the result could not be interpreted for man. I have a view that all these agents could be transmitted provided a large enough dose by appropriate routes was given and the animals kept long enough. Until the mechanisms of the species barrier are more clearly understood it might be best to retain that hypothesis.


snip...


R. BRADLEY






BSE: TIME TO TAKE H.B. PARRY SERIOUSLY


If the scrapie agent is generated from ovine DNA and thence causes disease in other species, then perhaps, bearing in mind the possible role of scrapie in CJD of humans (Davinpour et al, 1985), scrapie and not BSE should be the notifiable disease. ...




Wednesday, February 16, 2011


IN CONFIDENCE


SCRAPIE TRANSMISSION TO CHIMPANZEES


IN CONFIDENCE






Sunday, April 18, 2010


SCRAPIE AND ATYPICAL SCRAPIE TRANSMISSION STUDIES A REVIEW 2010






Monday, April 25, 2011


Experimental Oral Transmission of Atypical Scrapie to Sheep


Volume 17, Number 5-May 2011






1: J Infect Dis 1980 Aug;142(2):205-8


Oral transmission of kuru, Creutzfeldt-Jakob disease, and scrapie to nonhuman primates.


Gibbs CJ Jr, Amyx HL, Bacote A, Masters CL, Gajdusek DC.


Kuru and Creutzfeldt-Jakob disease of humans and scrapie disease of sheep and goats were transmitted to squirrel monkeys (Saimiri sciureus) that were exposed to the infectious agents only by their nonforced consumption of known infectious tissues. The asymptomatic incubation period in the one monkey exposed to the virus of kuru was 36 months; that in the two monkeys exposed to the virus of Creutzfeldt-Jakob disease was 23 and 27 months, respectively; and that in the two monkeys exposed to the virus of scrapie was 25 and 32 months, respectively. Careful physical examination of the buccal cavities of all of the monkeys failed to reveal signs or oral lesions. One additional monkey similarly exposed to kuru has remained asymptomatic during the 39 months that it has been under observation.


snip...


The successful transmission of kuru, Creutzfeldt-Jakob disease, and scrapie by natural feeding to squirrel monkeys that we have reported provides further grounds for concern that scrapie-infected meat may occasionally give rise in humans to Creutzfeldt-Jakob disease.


PMID: 6997404






12/10/76


AGRICULTURAL RESEARCH COUNCIL REPORT OF THE ADVISORY COMMITTE ON SCRAPIE


Office Note CHAIRMAN: PROFESSOR PETER WILDY


snip...


A The Present Position with respect to Scrapie A] The Problem Scrapie is a natural disease of sheep and goats. It is a slow and inexorably progressive degenerative disorder of the nervous system and it ia fatal. It is enzootic in the United Kingdom but not in all countries. The field problem has been reviewed by a MAFF working group (ARC 35/77). It is difficult to assess the incidence in Britain for a variety of reasons but the disease causes serious financial loss; it is estimated that it cost Swaledale breeders alone $l.7 M during the five years 1971-1975. A further inestimable loss arises from the closure of certain export markets, in particular those of the United States, to British sheep. It is clear that scrapie in sheep is important commercially and for that reason alone effective measures to control it should be devised as quickly as possible. Recently the question has again been brought up as to whether scrapie is transmissible to man. This has followed reports that the disease has been transmitted to primates.


One particularly lurid speculation (Gajdusek 1977) conjectures that the agents of scrapie, kuru, Creutzfeldt-Jakob disease and transmissible encephalopathy of mink are varieties of a single "virus". The U.S. Department of Agriculture concluded that it could "no longer justify or permit scrapie-blood line and scrapie-exposed sheep and goats to be processed for human or animal food at slaughter or rendering plants" (ARC 84/77)" The problem is emphasised by the finding that some strains of scrapie produce lesions identical to the once which characterise the human dementias" Whether true or not. the hypothesis that these agents might be transmissible to man raises two considerations. First, the safety of laboratory personnel requires prompt attention. Second, action such as the "scorched meat" policy of USDA makes the solution of the acrapie problem urgent if the sheep industry is not to suffer grievously.


snip...


76/10.12/4.6






Nature. 1972 Mar 10;236(5341):73-4.


Transmission of scrapie to the cynomolgus monkey (Macaca fascicularis).


Gibbs CJ Jr, Gajdusek DC. Nature 236, 73 - 74 (10 March 1972); doi:10.1038/236073a0


Transmission of Scrapie to the Cynomolgus Monkey (Macaca fascicularis)


C. J. GIBBS jun. & D. C. GAJDUSEK National Institute of Neurological Diseases and Stroke, National Institutes of Health, Bethesda, Maryland


SCRAPIE has been transmitted to the cynomolgus, or crab-eating, monkey (Macaca fascicularis) with an incubation period of more than 5 yr from the time of intracerebral inoculation of scrapie-infected mouse brain. The animal developed a chronic central nervous system degeneration, with ataxia, tremor and myoclonus with associated severe scrapie-like pathology of intensive astroglial hypertrophy and proliferation, neuronal vacuolation and status spongiosus of grey matter. The strain of scrapie virus used was the eighth passage in Swiss mice (NIH) of a Compton strain of scrapie obtained as ninth intracerebral passage of the agent in goat brain, from Dr R. L. Chandler (ARC, Compton, Berkshire).




Sunday, March 28, 2010


Nor-98 atypical Scrapie, atypical BSE, spontaneous TSE, trade policy, sound science ?






Monday, November 30, 2009


USDA AND OIE COLLABORATE TO EXCLUDE ATYPICAL SCRAPIE NOR-98 ANIMAL HEALTH CODE






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


Chronic Wasting Disease CWD


Friday, October 26, 2012


CHRONIC WASTING DISEASE CWD PENNSYLVANIA GAME FARMS, URINE ATTRACTANT PRODUCTS, BAITING, AND MINERAL LICKS








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




Monday, March 26, 2012


CANINE SPONGIFORM ENCEPHALOPATHY: A NEW FORM OF ANIMAL PRION DISEASE


OR-09 15:10 - 15:25


CANINE SPONGIFORM ENCEPHALOPATHY: A NEW FORM OF ANIMAL PRION DISEASE




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


PO-081: Chronic wasting disease in the cat— Similarities to feline spongiform encephalopathy (FSE)










PO-081: Chronic wasting disease in the cat— Similarities to feline spongiform encephalopathy (FSE)






Thursday, May 31, 2012


CHRONIC WASTING DISEASE CWD PRION2012 Aerosol, Inhalation transmission, Scrapie, cats, species barrier, burial, and more




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




SPORADIC CJD, FFI, AND GSS ???


AUGUST 2012 CJD type determination pending, sporadic cases include 16 cases of sporadic Fatal Insomnia (sFI) and 42 cases of Variably Protease-Sensitive Prionopathy (VPSPr) ???


really ???


see ;


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 was acquired in the United Kingdom in one case and in Saudi Arabia in the other case;


*** 5 Includes 8 cases in which the diagnosis is pending, and 18 inconclusive cases;


*** 6 Includes 10 (9 from 2012) cases with type determination pending in which the diagnosis of vCJD has been excluded.


*** The Sporadic cases include 16 cases of sporadic Fatal Insomnia (sFI) and 42 cases of Variably Protease-Sensitive Prionopathy (VPSPr) and 2224 cases of sporadic Creutzfeldt-Jakob disease (sCJD).






Tuesday, June 26, 2012


Creutzfeldt Jakob Disease Human TSE report update North America, Canada, Mexico, and USDA PRION UNIT as of May 18, 2012


type determination pending Creutzfeldt Jakob Disease (tdpCJD), is on the rise in Canada and the USA






a review of the science on the cjd follies, which has been driven by the industry all along $$$




Wednesday, October 27, 2010


A novel variant of human disease with a protease-sensitive prion protein and heterozygosity methionine/valine at codon 129: Case report


snip...


Genetic findings


No mutations were found in the open reading frame after sequencing the prion protein gene (PRNP). A heterozygosis methionine valine (MV) was observed in codon 129.


snip...






Thursday, July 10, 2008


A Novel Human Disease with Abnormal Prion Protein Sensitive to Protease update July 10, 2008


Although several subjects had family histories of dementia, no mutations were found in the PrP gene open reading frame.






Thursday, July 10, 2008


A New Prionopathy update July 10, 2008






***+++***


Thursday, July 10, 2008


A Novel Human Disease with Abnormal Prion Protein Sensitive to Protease update July 10, 2008 Friday, June 20, 2008






Here we go folks. AS predicted. THIS JUST OUT !


Tuesday, August 03, 2010


Variably protease-sensitive prionopathy: A new sporadic disease of the prion protein






PRIONBALONEY PrP$$$


Monday, August 9, 2010


Variably protease-sensitive prionopathy: A new sporadic disease of the prion protein or just more PRIONBALONEY ?


snip...see full text ;










O.K. let's compare some recent cases of this prionpathy in other countries besides Gambetti's first 10 recently, that he claims is a spontaneous event, from a genetic disorder, that is not genetic, but sporadic, that is related to no animal TSE in North America, or the world. ...






Wednesday, October 27, 2010


A novel variant of human disease with a protease-sensitive prion protein and heterozygosity methionine/valine at codon 129: Case report






Sunday, August 09, 2009


CJD...Straight talk with...James Ironside...and...Terry Singeltary... 2009






Tuesday, August 18, 2009


BSE-The Untold Story - joe gibbs and singeltary 1999 – 2009






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


The familial mutations, Gajdusek proposed, lowered the barrier to such accidental conversion. "Thus," he wrote in 1996, "with these mutations, this ordinarily rare event becomes a ... dominant inherited trait." But Weissmann's qualification still remained to be refuted: the mutations might simply allow easier entry to a lurking virus. ...page 202 Deadly Feast


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




something to think about for sure.


but i interpret this as (1st not the gold standard, just my opinion;-), as because of certain gene mutations, one or a family, would be more susceptible to the many different strains of TSE, and the many different proven routes and sources, (which will cause different symptoms, different incubation periods from onset of clinical symptoms to death, different parts of the brain infected, etc.). in other words, it's NOT the gene mutation that CAUSES the disease, but the fact that it makes you more SUSCEPTIBLE, to the TSEs from the surrounding environment, and PLUS accumulation, i think this plays a critical role. maybe there is a one dose scenario, but i think there is more of the 'accumulators' that go clinical, than the 'one dose'. and what is the threshold to sub-clinical to clinical ?




anyway, just pondering out loud here.


also, for anyone interested, there are some studies with links to follow here ;






Thursday, June 21, 2012


Clinical and Pathologic Features of H-Type Bovine Spongiform Encephalopathy Associated with E211K Prion Protein Polymorphism






let's take a closer look at this new prionpathy or prionopathy, and then let's look at the g-h-BSEalabama mad cow.




This new prionopathy in humans? the genetic makeup is IDENTICAL to the g-h-BSEalabama mad cow, the only _documented_ mad cow in the world to date like this, ......wait, it get's better. this new prionpathy is killing young and old humans, with LONG DURATION from onset of symptoms to death, and the symptoms are very similar to nvCJD victims, OH, and the plaques are very similar in some cases too, bbbut, it's not related to the g-h-BSEalabama cow, WAIT NOW, it gets even better, the new human prionpathy that they claim is a genetic TSE, has no relation to any gene mutation in that family. daaa, ya think it could be related to that mad cow with the same genetic make-up ??? there were literally tons and tons of banned mad cow protein in Alabama in commerce, and none of it transmitted to cows, and the cows to humans there from ??? r i g h t $$$


ALABAMA MAD COW g-h-BSEalabama


In this study, we identified a novel mutation in the bovine prion protein gene (Prnp), called E211K, of a confirmed BSE positive cow from Alabama, United States of America. This mutation is identical to the E200K pathogenic mutation found in humans with a genetic form of CJD. This finding represents the first report of a confirmed case of BSE with a potential pathogenic mutation within the bovine Prnp gene. We hypothesize that the bovine Prnp E211K mutation most likely has caused BSE in "the approximately 10-year-old cow" carrying the E221K mutation.












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: maf12@cam.ac.uk Jürgen A. Richt College of Veterinary Medicine, Kansas State University, K224B Mosier Hall, Manhattan, Kansas 66506-5601, USA


NATURE|Vol 457|26 February 2009






Saturday, August 14, 2010


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


(see mad cow feed in COMMERCE IN ALABAMA...TSS)








Sunday, February 12, 2012


National Prion Disease Pathology Surveillance Center Cases Examined1 (August 19, 2011) including Texas




kind regards, terry


Wednesday, March 28, 2012


VARIABLY PROTEASE-SENSITVE PRIONOPATHY IS TRANSMISSIBLE, price of prion poker goes up again $








P.9.21


Molecular characterization of BSE in Canada


Jianmin Yang1, Sandor Dudas2, Catherine Graham2, Markus Czub3, Tim McAllister1, Stefanie Czub1 1Agriculture and Agri-Food Canada Research Centre, Canada; 2National and OIE BSE Reference Laboratory, Canada; 3University of Calgary, Canada


Background: Three BSE types (classical and two atypical) have been identified on the basis of molecular characteristics of the misfolded protein associated with the disease. To date, each of these three types have been detected in Canadian cattle.


Objectives: This study was conducted to further characterize the 16 Canadian BSE cases based on the biochemical properties of there associated PrPres. Methods: Immuno-reactivity, molecular weight, glycoform profiles and relative proteinase K sensitivity of the PrPres from each of the 16 confirmed Canadian BSE cases was determined using modified Western blot analysis.


Results: Fourteen of the 16 Canadian BSE cases were C type, 1 was H type and 1 was L type. The Canadian H and L-type BSE cases exhibited size shifts and changes in glycosylation similar to other atypical BSE cases. PK digestion under mild and stringent conditions revealed a reduced protease resistance of the atypical cases compared to the C-type cases. N terminal- specific antibodies bound to PrPres from H type but not from C or L type. The C-terminal-specific antibodies resulted in a shift in the glycoform profile and detected a fourth band in the Canadian H-type BSE.


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


*** It also suggests a similar cause or source for atypical BSE in these countries.






what about that ALABAMA MAD COW, AND MAD COW FEED THERE FROM IN THAT STATE ???


Saturday, August 14, 2010


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


*** (see mad cow feed in COMMERCE IN ALABAMA...TSS)


BANNED MAD COW FEED IN COMMERCE IN ALABAMA


Date: September 6, 2006 at 7:58 am PST PRODUCT


a) EVSRC Custom dairy feed, Recall # V-130-6;


b) Performance Chick Starter, Recall # V-131-6;


c) Performance Quail Grower, Recall # V-132-6;


d) Performance Pheasant Finisher, Recall # V-133-6.


CODE None RECALLING FIRM/MANUFACTURER Donaldson & Hasenbein/dba J&R Feed Service, Inc., Cullman, AL, by telephone on June 23, 2006 and by letter dated July 19, 2006. Firm initiated recall is complete.


REASON


Dairy and poultry feeds were possibly contaminated with ruminant based protein.


VOLUME OF PRODUCT IN COMMERCE 477.72 tons


DISTRIBUTION AL


______________________________




PRODUCT Bulk custom dairy pre-mixes,


Recall # V-120-6 CODE None RECALLING FIRM/MANUFACTURER Ware Milling Inc., Houston, MS, by telephone on June 23, 2006. Firm initiated recall is complete. REASON Possible contamination of dairy animal feeds with ruminant derived meat and bone meal.


VOLUME OF PRODUCT IN COMMERCE 350 tons


DISTRIBUTION AL and MS


______________________________


PRODUCT


a) Tucker Milling, LLC Tm 32% Sinking Fish Grower, #2680-Pellet, 50 lb. bags, Recall # V-121-6;


b) Tucker Milling, LLC #31120, Game Bird Breeder Pellet, 50 lb. bags, Recall # V-122-6;


c) Tucker Milling, LLC #31232 Game Bird Grower, 50 lb. bags, Recall # V-123-6;


d) Tucker Milling, LLC 31227-Crumble, Game Bird Starter, BMD Medicated, 50 lb bags, Recall # V-124-6;


e) Tucker Milling, LLC #31120, Game Bird Breeder, 50 lb bags, Recall # V-125-6;


f) Tucker Milling, LLC #30230, 30 % Turkey Starter, 50 lb bags, Recall # V-126-6;


g) Tucker Milling, LLC #30116, TM Broiler Finisher, 50 lb bags, Recall # V-127-6


CODE All products manufactured from 02/01/2005 until 06/20/2006 RECALLING FIRM/MANUFACTURER Recalling Firm: Tucker Milling LLC, Guntersville, AL, by telephone and visit on June 20, 2006, and by letter on June 23, 2006. Manufacturer: H. J. Baker and Brothers Inc., Stamford, CT. Firm initiated recall is ongoing.


REASON Poultry and fish feeds which were possibly contaminated with ruminant based protein were not labeled as "Do not feed to ruminants".


VOLUME OF PRODUCT IN COMMERCE 7,541-50 lb bags


DISTRIBUTION AL, GA, MS, and TN


END OF ENFORCEMENT REPORT FOR AUGUST 9, 2006


###




Subject: MAD COW FEED RECALL AL AND FL VOLUME OF PRODUCT IN COMMERCE 125 TONS Products manufactured from 02/01/2005 until 06/06/2006


Date: August 6, 2006 at 6:16 pm PST PRODUCT


a) CO-OP 32% Sinking Catfish, Recall # V-100-6;


b) Performance Sheep Pell W/Decox/A/N, medicated, net wt. 50 lbs, Recall # V-101-6;


c) Pro 40% Swine Conc Meal -- 50 lb, Recall # V-102-6;


d) CO-OP 32% Sinking Catfish Food Medicated, Recall # V-103-6;


e) "Big Jim's" BBB Deer Ration, Big Buck Blend, Recall # V-104-6;


f) CO-OP 40% Hog Supplement Medicated Pelleted, Tylosin 100 grams/ton, 50 lb. bag, Recall # V-105-6;


g) Pig Starter Pell II, 18% W/MCDX Medicated 282020, Carbadox -- 0.0055%, Recall # V-106-6;


h) CO-OP STARTER-GROWER CRUMBLES, Complete Feed for Chickens from Hatch to 20 Weeks, Medicated, Bacitracin Methylene Disalicylate, 25 and 50 Lbs, Recall # V-107-6;


i) CO-OP LAYING PELLETS, Complete Feed for Laying Chickens, Recall # 108-6;


j) CO-OP LAYING CRUMBLES, Recall # V-109-6;


k) CO-OP QUAIL FLIGHT CONDITIONER MEDICATED, net wt 50 Lbs, Recall # V-110-6;


l) CO-OP QUAIL STARTER MEDICATED, Net Wt. 50 Lbs, Recall # V-111-6;


m) CO-OP QUAIL GROWER MEDICATED, 50 Lbs, Recall # V-112-6 CODE


Product manufactured from 02/01/2005 until 06/06/2006


RECALLING FIRM/MANUFACTURER Alabama Farmers Cooperative, Inc., Decatur, AL, by telephone, fax, email and visit on June 9, 2006. FDA initiated recall is complete.


REASON Animal and fish feeds which were possibly contaminated with ruminant based protein not labeled as "Do not feed to ruminants".


VOLUME OF PRODUCT IN COMMERCE 125 tons


DISTRIBUTION AL and FL


END OF ENFORCEMENT REPORT FOR AUGUST 2, 2006


###




MAD COW FEED RECALL USA EQUALS 10,878.06 TONS NATIONWIDE Sun Jul 16, 2006 09:22 71.248.128.67


RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINE -- CLASS II


______________________________


PRODUCT


a) PRO-LAK, bulk weight, Protein Concentrate for Lactating Dairy Animals, Recall # V-079-6;


b) ProAmino II, FOR PREFRESH AND LACTATING COWS, net weight 50lb (22.6 kg), Recall # V-080-6;


c) PRO-PAK, MARINE & ANIMAL PROTEIN CONCENTRATE FOR USE IN ANIMAL FEED, Recall # V-081-6;


d) Feather Meal, Recall # V-082-6 CODE


a) Bulk


b) None


c) Bulk


d) Bulk


RECALLING FIRM/MANUFACTURER H. J. Baker & Bro., Inc., Albertville, AL, by telephone on June 15, 2006 and by press release on June 16, 2006. Firm initiated recall is ongoing.


REASON


Possible contamination of animal feeds with ruminent derived meat and bone meal.


VOLUME OF PRODUCT IN COMMERCE 10,878.06 tons


DISTRIBUTION Nationwide


END OF ENFORCEMENT REPORT FOR July 12, 2006


###




Saturday, July 23, 2011


CATTLE HEADS WITH TONSILS, BEEF TONGUES, SPINAL CORD, SPECIFIED RISK MATERIALS (SRM's) AND PRIONS, AKA MAD COW DISEASE






Saturday, November 6, 2010


TAFS1 Position Paper on Position Paper on Relaxation of the Feed Ban in the EU Berne, 2010 TAFS


INTERNATIONAL FORUM FOR TRANSMISSIBLE ANIMAL DISEASES AND FOOD SAFETY a non-profit Swiss Foundation






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






USDA TRIPLE BSE MAD COW FIREWALL, SRM, FEED, AND SURVEILLANCE


2012


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


Second threat


snip...






MAD COW USDA ATYPICAL L-TYPE BASE BSE, the rest of the story...


***Oral Transmission of L-type Bovine Spongiform Encephalopathy in Primate Model






***Infectivity in skeletal muscle of BASE-infected cattle






***feedstuffs- It also suggests a similar cause or source for atypical BSE in these countries.






***Also, a link is suspected between atypical BSE and some apparently sporadic cases of Creutzfeldt-Jakob disease in humans.






The present study demonstrated successful intraspecies transmission of H-type BSE to cattle and the distribution and immunolabeling patterns of PrPSc in the brain of the H-type BSE-challenged cattle. TSE agent virulence can be minimally defined by oral transmission of different TSE agents (C-type, L-type, and H-type BSE agents) [59]. Oral transmission studies with H-type BSEinfected cattle have been initiated and are underway to provide information regarding the extent of similarity in the immunohistochemical and molecular features before and after transmission.


In addition, the present data will support risk assessments in some peripheral tissues derived from cattle affected with H-type BSE.






Thursday, June 21, 2012


Clinical and Pathologic Features of H-Type Bovine Spongiform Encephalopathy Associated with E211K Prion Protein Polymorphism


Justin J. Greenlee1*, Jodi D. Smith1, M. Heather West Greenlee2, Eric M. Nicholson1


1 National Animal Disease Center, United States Department of Agriculture, Agricultural Research Service, Ames, Iowa, United States of America, 2 Iowa State University, Ames, Iowa, United States of America


Abstract


The majority of bovine spongiform encephalopathy (BSE) cases have been ascribed to the classical form of the disease. Htype and L-type BSE cases have atypical molecular profiles compared to classical BSE and are thought to arise spontaneously. However, one case of H-type BSE was associated with a heritable E211K mutation in the prion protein gene. The purpose of this study was to describe transmission of this unique isolate of H-type BSE when inoculated into a calf of the same genotype by the intracranial route. Electroretinograms were used to demonstrate preclinical deficits in retinal function, and optical coherence tomography was used to demonstrate an antemortem decrease in retinal thickness. The calf rapidly progressed to clinical disease (9.4 months) and was necropsied. Widespread distribution of abnormal prion protein was demonstrated within neural tissues by western blot and immunohistochemistry. While this isolate is categorized as BSE-H due to a higher molecular mass of the unglycosylated PrPSc isoform, a strong labeling of all 3 PrPSc bands with monoclonal antibodies 6H4 and P4, and a second unglycosylated band at approximately 14 kDa when developed with antibodies that bind in the C-terminal region, it is unique from other described cases of BSE-H because of an additional band 23 kDa demonstrated on western blots of the cerebellum. This work demonstrates that this isolate is transmissible, has a BSE-H phenotype when transmitted to cattle with the K211 polymorphism, and has molecular features that distinguish it from other cases of BSE-H described in the literature.


snip...


Most significantly it must be determined if the molecular phenotype of this cattle TSE remains stable when transmitted to cattle without the E211K polymorphism as several other isolates of atypical BSE have been shown to adopt a molecular profile consistent with classical BSE after passage in transgenic mice expressing bovine PrPC [40] or multiple passages in wild type mice [23]. Results of ongoing studies, namely passage of the E211K Htype isolate into wild-type cattle, will lend further insight into what role, if any, genetic and sporadic forms of BSE may have played in the origins of classical BSE. Atypical cases presumably of spontaneous or, in the case of E211K BSE-H, genetic origins highlight that it may not be possible to eradicate BSE entirely and that it would be hazardous to remove disease control measures such as prohibiting the feeding of meat and bone meal to ruminants.








Saturday, May 26, 2012


Are USDA assurances on mad cow case 'gross oversimplification'?


SNIP...


What irks many scientists is the USDA’s April 25 statement that the rare disease is “not generally associated with an animal consuming infected feed.”


The USDA’s conclusion is a “gross oversimplification,” said Dr. Paul Brown, one of the world’s experts on this type of disease who retired recently from the National Institutes of Health. "(The agency) has no foundation on which to base that statement.”


“We can’t say it’s not feed related,” agreed Dr. Linda Detwiler, an official with the USDA during the Clinton Administration now at Mississippi State.


In the May 1 email to me, USDA’s Cole backed off a bit. “No one knows the origins of atypical cases of BSE,” she said


The argument about feed is critical because if feed is the cause, not a spontaneous mutation, the California cow could be part of a larger outbreak.


SNIP...






Saturday, November 6, 2010


TAFS1 Position Paper on Position Paper on Relaxation of the Feed Ban in the EU


Berne, 2010 TAFS INTERNATIONAL FORUM FOR TRANSMISSIBLE ANIMAL DISEASES AND FOOD SAFETY a non-profit Swiss Foundation






Archive Number 20101206.4364 Published Date 06-DEC-2010 Subject PRO/AH/EDR>


Prion disease update 2010 (11) PRION DISEASE UPDATE 2010 (11)






Saturday, October 6, 2012


TRANSMISSION, DIFFERENTIATION, AND PATHOBIOLOGY OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHIES 2011 Annual Report






CJD UPDATE USA




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 was acquired in the United Kingdom in one case and in Saudi Arabia in the other case;


*** 5 Includes 8 cases in which the diagnosis is pending, and 18 inconclusive cases;


*** 6 Includes 10 (9 from 2012) cases with type determination pending in which the diagnosis of vCJD has been excluded.


*** The Sporadic cases include 16 cases of sporadic Fatal Insomnia (sFI) and 42 cases of Variably Protease-Sensitive Prionopathy (VPSPr) and 2224 cases of sporadic Creutzfeldt-Jakob disease (sCJD).






Tuesday, June 26, 2012


Creutzfeldt Jakob Disease Human TSE report update North America, Canada, Mexico, and USDA PRION UNIT as of May 18, 2012


type determination pending Creutzfeldt Jakob Disease (tdpCJD), is on the rise in Canada and the USA






DEEP THROAT TO TSS 2000-2001 (take these old snips of emails with how ever many grains of salt you wish. ...tss)


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 a least a million bovine tested as soon as possible and agressively seeking this disease. The big players are coming out of the woodwork 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 a great 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.)






U.S.A. 50 STATE BSE MAD COW CONFERENCE CALL Jan. 9, 2001


Subject: BSE--U.S. 50 STATE CONFERENCE CALL Jan. 9, 2001






Friday, August 24, 2012


Iatrogenic prion diseases in humans: an update








Monday, July 23, 2012


The National Prion Disease Pathology Surveillance Center July 2012






the costly price of politics and TSE prion disease...




Monday, August 20, 2012


CASE REPORTS CREUTZFELDT-JAKOB DISEASE: AN UNDER-RECOGNIZED CAUSE OF DEMENTIA






Saturday, October 13, 2012


On the issue of transmissibility of Alzheimer disease: A critical review






layperson


TSS