Mortality of Individuals With PRNP Variants Associated With Prion Disease in the United States, 1998–2024 Singeltary Review 2026  August 26, 2026

 Mortality of Individuals With PRNP Variants Associated With Prion Disease in the United States, 1998–2024 Singeltary Review 2026  August 26, 2026

Open Access

Mortality of Individuals With PRNP Variants Associated With Prion Disease in the United States, 1998–2024

Yuan Lian, Keisi Kotobelli, Katie Glisic, Daniel A. Sprague, Sonia M. Vallabh, Brian S. Appleby ericminikel@gmail.com, and Eric Vallabh Minikel

AUTHORS INFO & AFFILIATIONS October 2026 issue 12 (5) e200423

https://doi.org/10.1212/NXG.0000000000200423

Abstract

Background and Objectives

To characterize the survival of individuals with pathogenic PRNP variants—including to estimate annual hazards, to judge the accuracy of previously reported survival data, and to evaluate the utility of public record searches in determining vital status. Methods

In this single-center cohort study, we gathered data on individuals who received positive antemortem PRNP genetic tests at the US National Prion Disease Pathology Surveillance Center (NPDPSC), including both diagnostic tests in symptomatic individuals, and predictive tests in asymptomatic individuals. Genetic test and autopsy results were queried from the NPDPSC database, and public record searches were conducted using online tools. Results

Four hundred four individuals received positive genetic test results. Of 206 cases symptomatic at the time of genetic testing, 188 are likely now deceased based on typical disease duration for their genetic variants. Combined autopsy and public record searches in combination confirmed 174 of these deaths, for an estimated 92.6% sensitivity. We evaluated the age-dependent penetrance of the reportedly highly penetrance variants D178N and E200K and the reportedly low-penetrance variant V210I. Among 99 initially asymptomatic individuals with the pathogenic E200K variant, more than 936 person-years of follow-up, 18 deaths were observed, significantly fewer than 27.4 expected according to life tables based on retrospective data. The age-dependent penetrance of E200K calculated from these longitudinal data was significantly lower than that from retrospective data, with 69% penetrance by age 80 and a median age at death of 75. For the pathogenic D178N variant, the median age at death was 57, which was numerically later, but not significantly different from, that seen in retrospective data. For V210I, just 2 deaths occurred, both after age 90, consistent with minimal penetrance. Discussion

Our data support high penetrance of PRNP D178N and E200K variants and low penetrance of V210I. For E200K, the age at onset distribution appears to be shifted slightly later, and lifetime risk slightly lower, than previously reported. Autopsy data and public death records in combination were sensitive and concordant for determining long-term outcomes, but additional prospective data should be gathered to support future preventive trials.

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Discussion

The penetrance and age at onset of PRNP variants have previously been classified based on the presence of Mendelian segregation and/or de novo variation in the literature, comparison of allele frequency in cases and controls, and aggregation of age at onset data chiefly as “snapshots” of symptomatic or deceased cases reported to surveillance centers worldwide. All of these methods are useful but are imperfect and subject to ascertainment bias. Individuals are generally more likely to appear in data sets when they develop disease (as opposed to never becoming sick) and are diagnosed, which may be a function of their age (better ascertainment in younger individuals) and their year of onset (improvements in surveillance and diagnosis over time3,4). Ascertainment bias can sometimes lead to surprising results, such as the false appearance of genetic anticipation.5,6

The ascertainment bias in snapshot data, wherein individuals are usually discovered only on presentation with active disease, is difficult to correct for, because the number of individuals who either never presented symptoms, or were not diagnosed, cannot be empirically measured. A more ideal way to determine the age-dependent penetrance of genetic variants would be to longitudinally follow individuals with disease-causing variants and determine the proportion that succumb to disease in each year of life. This method carries its own ascertainment bias, specifically, that individuals only enter the data set at the age at which they undergo genetic testing, but importantly, our method corrects for this bias in delayed entry to the at-risk population.

In this study, we examined the survival of individuals with pathogenic PRNP variants identified when still asymptomatic, capturing 1,836 person-years of follow-up on 198 individuals. Among the most common variants in our data set were P102L, D178N, and E200K, all of which are reported to be highly penetrant and which comprise 85% of cases with highly penetrant variants worldwide, as well as V210I, which is the most common low-penetrance variant in a global data set.7

The PRNP E200K variant is generally believed to cause genetic prion disease with high penetrance, based on its segregation with disease in many multigenerational families, the high proportion of cases with a positive family history, and its dramatic enrichment in prion disease cases over controls.1,7,8 Most reports on E200K have estimated >90% penetrance by age 80.2,5,9,10 Two reports argued for a lower penetrance, ∼60%, but in small cohorts lacking any public data set or any detailed description of the methods used for ascertainment or analysis.11,12 Among 99 E200K individuals examined in the longitudinal data set here, 18 died of definite or possible prion disease, with a median survival of 75 and estimated 69% penetrance by age 80. This differed significantly from the snapshot survival data, which would have predicted 27 deaths among the same individuals, with median survival of 62, and 96% penetrance by age 80. Our data still support classification of E200K as highly penetrant, but may point to a slightly more optimistic distribution of ages at onset. The D178N variant has an earlier age at onset and an even higher proportion of cases with positive family history, perhaps suggesting less opportunity for ascertainment bias to cloud the snapshot data. Indeed, there was no significant difference in the survival of D178N individuals in our data set compared with snapshot data: the median survival was slightly later—58 vs 52—but given the small number of deaths (6 deaths observed compared with 7 expected), this was within sampling variance. Our data support classification of D178N as highly penetrant.

Families with V210I generally do not exhibit multiplex segregation, only a minority (12%) of cases have a positive family history, and comparison of case and control allele frequency yielded a point estimate of 8% lifetime risk.7,8 We observed only 2 deaths past age 90 among V210I individuals, which may not have been due to prion disease (see below), consistent with low penetrance.

The above findings are subject to several biases and caveats. We assumed that individuals for whom we were unable to locate public records of death, such as online obituaries, are still alive. When we compared results of public record searches against autopsy data as the gold standard, we found that public record searches were 98.2% sensitive at detecting deaths. Autopsy cases could represent a biased sample, however. When we instead use as our gold standard the individuals who underwent diagnostic testing long enough ago that they are likely deceased based on disease duration for their variants, and we then assess the combined sensitivity of autopsy plus public record searches, we obtain a sensitivity of just 92.6%. To the extent that we erroneously assumed some individuals to be alive who actually died of prion disease, our estimates may be overly optimistic. On the other hand, we also assumed that deaths identified through public record searches without autopsy were due to prion disease. In fact, particularly for E200K, the range of ages at onset extends into the end of the natural human lifespan. If some deaths were not due to prion disease, then our estimates may be overly pessimistic. The above 2 limitations act in opposing directions: our imperfect sensitivity detect deaths may mean true hazards are higher than we estimate, while our assumption that all deaths are due to prion disease may mean true hazards are lower than we estimate. We explored the effect of these limitations through sensitivity tests. Under the most extreme set of assumptions in either direction, our study would still support the conclusion that E200K has a somewhat later age at onset and lower penetrance than previously believed, while still being highly penetrant.

Diagnosis and ascertainment of older individuals with prion disease appear to be improving over time. For example, UK surveillance data indicate that in sporadic prion disease, the measured annual incidence in older individuals, particularly age 80+, was approximately double in 2010–2016 what it was in 1995–2004.13 CSF diagnostic tests for prion disease appear to be ordered less often in the elderly, and brain MRI is less sensitive to prion disease in older individuals than in younger individuals.14 Thus, it is possible that prion disease remains underdiagnosed in older age groups. Our simulations suggest, however, that underascertainment of older disease onsets in our snapshot data is not sufficient to explain the younger median onset calculated from those data compared with the longitudinal data set.

In a previously reported UK genetic testing data set,15 of 42 individuals who tested positive for a PRNP variant, 7 of 10 who went on to become symptomatic did so within 1 year of testing, a significant excess over expectation. This was interpreted to suggest that some individuals seek predictive genetic testing due to a correct suspicion of initial symptoms of disease. This might introduce bias in the estimation of annual hazards after predictive testing. By contrast, only 3 deaths in our data set occurred shortly after genetic testing (meaning, within the 90th percentile of disease duration for their PRNP variant). This difference might be attributable to the composition of genetic variants represented in the 2 cohorts. Slowly progressive variants such as P102L, A117V, and 6-OPRI, which are overrepresented in the United Kingdom, are more likely to have an extended prodrome as well as more preserved cognition in the earlier stage of the illness, enabling these individuals to seek out predictive genetic testing while possibly experiencing prodromal symptoms. This US cohort consists primarily of rapidly progressive variants, especially E200K, similar to the world average.7

Our study has limitations. No single source of data exhaustively captures all deaths, so our estimate of combined sensitivity is imperfect. We also lacked a perfect gold standard for individuals still alive; thus, we cannot assess the specificity of public record searches. We did not use tokenization16 or other privacy-preserving record linkage methods to leverage additional sources of vital status information. Only 32 deaths occurred among initially asymptomatic individuals, still a small number from which to estimate hazards. We relied on public obituaries, which are common for ordinary people in the United States, but customs may differ globally. Thus, the method we used here may not be generalizable worldwide. A strength is that most of our data are from E200K, the most common disease-causing PRNP variant worldwide, which means that our findings regarding age-dependent penetrance have worldwide generalizability.

The variable age at symptom onset in prion disease has implications for clinical trial design in asymptomatic at-risk individuals with PRNP variants. It has been shown numerically infeasible to statistically power pivotal preapproval clinical trials to demonstrate that a drug has delayed onset of disease in the at-risk population.2 Instead, one clinical development path for preventive drugs would require provisional approval on a biomarker end point such as CSF PrP.17,18 If initial approval of a preventive drug for at-risk people is based on a biomarker end point, then long-term survival might be assessed in a postmarketing study.19 Our findings may suggest a need to refine our estimates of annual hazards facing asymptomatic individuals with PRNP variants to facilitate such postmarketing studies. An important future direction will be the establishment of a prospective registry study in PRNP at-risk individuals. Asking for consent for contact next of kin as well as for tokenization would expand the set of methods available to minimize informative censoring and ensure that no participants are lost to follow-up. Such a study could gather additional prospective data in untreated individuals in advance of prevention trials while also providing an infrastructure for future postmarketing studies.

https://www.neurology.org/doi/10.1212/NXG.0000000000200423

This study demonstrates that the H-type BSE agent is transmissible by the oronasal route…Interestingly, one case of H-type BSE in the US was associated with an inherited mutation in the prion protein gene referred to as E211K.

https://www.ars.usda.gov/research/publications/publication/?seqNo115=395351

Cattle with the EK211 PRNP polymorphism are susceptible to the H-type bovine spongiform encephalopathy agent from either E211K or wild type donors after oronasal inoculation

Justin J. Greenleea, Eric D. Cassmanna, S. Jo Moorea,b, and M. Heather West Greenleec

aVirus and Prion Research Unit, National Animal Disease Center, ARS, United States Department of Agriculture, Ames, IA, USA; bOak Ridge Institute for Science and Education (ORISE), U.S. Department of Energy, Oak Ridge, TN, US; cDepartment of Biomedical Sciences, Iowa State University College of Veterinary Medicine, Ames, IA, US

Aims: In 2006, a case of H-type bovine spongiform encephalopathy (H-BSE) was reported in a cow with a previously unreported prion protein polymorphism (E211K). The E211K polymorphism is heritable and homologous to the E200K mutation in humans that is the most frequent PRNP mutation associated with familial Creutzfeldt-Jakob disease. Although the prevalence of the E211K polymorphism is low, cattle carrying the K211 allele develop H-type BSE with a rapid onset after experimental inoculation by the intracranial route. The purpose of this study was to investigate whether the agents of H-type BSE or H-type BSE associated with the E211K polymorphism transmit to wild type cattle or cattle with the K211 allele after oronasal exposure.

Material and Methods: Wild type (EE211) or heterozygous (EK211) cattle were oronasally inoculated with the H-BSE agent from either the US 2004 case (wild type donor; n = 3) or from the US 2006 case with the E211K polymorphism (n = 4). Cattle were observed daily throughout the course of the experiment for the development of clinical signs. When signs were noted, animals were euthanized and necropsied. Cattle were confirmed positive for abnormal BSE prions by enzyme immunoassay (EIA; Idexx HerdChek BSE Ag Test), anti-PrP immunohistochemistry (IHC) on brainstem, and microscopic examination for vacuolation.

Results: Three-out-of-four (75%) calves with the EK211 genotype developed clinical signs of H-BSE including inattentiveness, loss of body condition, weakness, ataxia, and muscle fasciculations and were euthanized. Two of the positive EK211 steers received H-BSE US 2004 inoculum (Incubation Period (IP): 59.3 and 72.3 months) while the other positive steer received the E211K H-BSE inoculum (IP: 49.7 months). EIA confirmed that abundant misfolded protein (O.D. 2.57–4.0) in the brainstem, and IHC demonstrated PrPScthroughout the brain. All wild type recipient cattle and a single EK211 steer remained asymptomatic for the duration of the experiment (approximately 7 years post-inoculation) and no abnormal prion protein was detected in these cattle by EIA.

Conclusions: This study demonstrates that the H-type BSE agent is transmissible by the oronasal route. Cattle with the EK211 genotype are oronasally susceptible to small doses of the H-BSE agent from either EK211 or EE211 (wild type) donors. Wild-type EE211 cattle remained asymptomatic for the duration of the experiment with this small dose (0.1 g) of inoculum. These results reinforce the need for ongoing surveillance for classical and atypical BSE to minimize the risk of potentially infectious tissues entering the animal or human food chains.

Funded by: US Department of Agriculture

Acknowledgement: This research was funded in its entirety by congressionally appropriated funds to the United States Department of Agriculture, Agricultural Research Service. The funders of the work did not influence study design, data collection and analysis, decision to publish, or preparation of the manuscript. This research was supported in part by an appointment to the Agricultural Research Service (ARS) Research Participation Program administered by the Oak Ridge Institute for Science and Education (ORISE) through an interagency agreement between the U.S. Department of Energy (DOE) and the U.S. Department of Agriculture (USDA). ORISE is managed by ORAU under DOE contract number DE-SC0014664. All opinions expressed in this paper are the author’s and do not necessarily reflect the policies and views of USDA, ARS, DOE, or ORAU/ORISE.

https://www.tandfonline.com/doi/full/10.1080/19336896.2022.2091286

Canadian 2021 H-type Bovine Spongiform Encephalopathy case associated with a novel E211K polymorphism in prion protein gene novel E211K polymorphism in prion protein gene

Waqas Tahir , Sandor Dudas , Renee Anderson , Jianmin Yang , Sarah Bogart , Kristina Santiago-Mateo, Yuanmu Fang & Roberta Quaghebeur

Pages 36-49 | Received 20 Feb 2025, Accepted 22 May 2025, Published online: 04 Aug 2025 Cite this article https://doi.org/10.1080/19336896.2025.2511933

ABSTRACT

Bovine Spongiform Encephalopathy (BSE) is a fatal neurodegenerative disease in cattle which can be either classical BSE (C-BSE) or atypical BSE (including H-BSE and L-BSE). Here, we report the results of our analyses of an H-BSE case found in Canada in 2021, indicating restriction of the pathological agent (PrPSc) mainly to the central nervous system with no or occasional weak involvement of peripheral tissues. Importantly, a non-synonymous mutation at codon 211 of the PRNP gene was detected and confirmed to be present as a germline mutation. This is the first case of BSE in Canada with a predisposing E211K mutation.

Introduction

The transmissible spongiform encephalopathies (TSEs) or prion diseases are fatal neurodegenerative diseases affecting both humans and animals [Citation1]. TSEs may occur sporadically, genetically or be acquired and are caused by a conformational change of a host encoded cellular prion protein (PrPC) into its pathological isoform called the scrapie prion protein (PrPSc) [Citation2]. This conformational shift to PrPSc leads to its deposition and aggregation in the central nervous system (CNS), resulting in neuronal loss and eventually death. Bovine spongiform encephalopathy (BSE) is a TSE of cattle. The first case of BSE (referred to as classical BSE; C-BSE) was identified in the UK in 1986 [Citation3] which evolved into an epidemic affecting more than 2 million cattle in the UK [Citation4]. Dietary exposure to C-BSE was later found to be the most likely cause of variant form of Creutzfeldt – Jacob disease (vCJD), an acquired form of prion disease in humans [Citation5,Citation6], indicating its zoonotic potential. The origin of the C-BSE epidemic in the UK was attributed to the recycling of TSE infected meat and bone meal (MBM) in animal feed [Citation7,Citation8]. This notion was supported by the reduction in the number of C-BSE cases worldwide when a ‘feed ban’ to eliminate MBM in animal feed was implemented in the affected countries. However, in 2004, two additional ‘atypical’ types of BSE, with different neuropathological and molecular characteristics from previous cases of C-BSE emerged in France and Italy [Citation9,Citation10]. These two atypical types of BSE were later designated as L-type (L-BSE) and H-type (H-BSE) based on the biochemical features of PrPSc (either lower or higher molecular weight un-glycosylated PrP in L-BSE and H-BSE respectively, compared to C-BSE) [Citation11,Citation12]. Since then, many other countries have reported atypical BSE cases. The exact cause of atypical BSE is unknown, however it is considered sporadic and linked to the old age of cattle since atypical BSE cases are mostly detected in cattle more than 8 years old with a few exceptions such as a Swiss H-BSE case in a cow of 6.5 years of age and a Japanese L-BSE case in a cow of 23 month old [Citation13,Citation14]. Moreover, a steady occurrence of atypical BSE cases, despite the feed ban, also favours their sporadic nature [Citation15].

The pathogenic PrPSc levels and distribution in various brain regions [Citation16] and peripheral tissues can be variable between C-BSE and atypical forms of BSE. Considering the public health concerns associated with BSE, it is highly pertinent to characterize any newly identified BSE case for the spatial pattern of PrPSc. Such information can help to determine if the distribution of the pathogenic PrPSc is the same as in previously reported cases or if there has been an evolution in prion strain biology and disease phenotype in newly identified BSE cases. Additionally, a set of genetic risk factors including a 23-bp indel in the promoter region, a 12-bp indel in intron 1 or an increase in the number of octa-peptide repeats (OPRs) on the N-terminal [Citation17–19] of the prion protein (PRNP) gene have also been identified to be associated with BSE cases. In 2006, the US reported a H-BSE case associated with a novel mutation in the bovine PRNP gene leading to a switch of Glutamic acid (E) to Lysine (K) at amino acid residue 211 of PrPC – referred to as the E211K mutation [Citation18]. The E211K mutation in the PRNP gene of cattle is analogous to the E200K mutation in the most prevalent genetic form of human TSE; genetic Creutzfeldt-Jakob’s disease (gCJD) [Citation20]. This E211K mutation in cattle was later found to be heritable [Citation21] and associated with an increased susceptibility to H-BSE [Citation22]. These findings underscore the importance to evaluate the presence of genetic risk factors in any newly identified case of BSE to aid in designing or modifying mitigation strategies to prevent future BSE cases.

Canada has reported 3 cases of atypical BSE (in addition to 17 cases of C-BSE) to date, with 1 case of L-BSE (in December 2007) and 2 cases of H-BSE (first one in July 2006 and the most recent in December 2021). There were no genetic risk factors or pathogenic mutations identified in the first two atypical BSE cases in Canada [Citation23]. Here, we report the results of our analyses of the most recent H-BSE case (Canadian BSE case 20), where we investigated, i) the spatial distribution of PrPSc to assess any changes in specified risk material (SRM), and ii) the genetic risk factors contributing to the sporadic disease onset in this case of H-BSE.

Materials and methods

Animal information

In December 2021, an 8-year-old beef cow of Red Poll breed from central Alberta, under the fallen stock stream of Canada and Alberta BSE surveillance programme, tested positive for BSE. The clinical presentation included loss of body weight, abnormal posture, ataxic and uncoordinated or exaggerated gait, and falling down or stumbling. The initial BSE surveillance testing of the brain stem from the cow was performed at a network laboratory in Alberta, Canada using World Organization for Animal Health (WOAH) approved rapid surveillance test for BSE. After repeating initial results, the sample was referred to the Canadian BSE Reference Lab; BSE-RL (Canadian Food Inspection Agency, National Center for Animal Disease, Lethbridge Laboratory, Lethbridge, Alberta, Canada) for confirmatory testing. After the sample tested positive at the BSE-RL, the obex from that cow was further analysed for the type of BSE by immunoblotting, presence of vacuolation by histopathology (H&E) and deposition of PrPSc by immunohistochemistry (IHC). After confirmation and typing, the carcass of the cow (Canadian BSE case 20) was brought to the BSE-RL for additional sample collection. Upon arrival at the BSE-RL, the carcass was decomposing, but all available tissues were harvested and analysed by IDEXX HerdChek (Bovine Spongiform Encephalopathy-Scrapie Antigen Test Kit, EIA), H&E and IHC for the presence and distribution of the pathological agent. Additionally, DNA from various tissues of the animal was also analysed to identify genetic variation in the PRNP gene.

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Identification of E211K mutation in the PRNP gene of the Canadian BSE case 20

To determine if the Canadian BSE case 20 was associated with any mutation in the PRNP gene, the PRNP gene was amplified from DNA extracted from brainstem and sequenced. The sequence alignment with a known bovine PRNP sequence showed that the Canadian BSE case 20 had a functional PRNP gene indicated by no mutation in either start or stop codon. An increase in the number of octa-peptide repeat regions on N- terminal of the PRNP gene has been shown to enhance susceptibility to BSE in transgenic rodent models [Citation17], however both alleles contained six octa-peptide repeats (OPRs), indicative of the wild type bovine genotype. However, sequence analysis of the coding region revealed a synonymous mutation at codon 192 (AAC/AAT > N192N) and a non-synonymous mutation at codon 211 (GAA/AAA > E211K) (Supplementary Figure S4 and Figure 4). The change in the nucleotide sequence at codon 211 from GAA to AAA resulted in a switch of amino acid sequence at codon 211 from glutamic acid (E) to lysine (K) amino acid. As this change in the nucleotide sequence from GAA to AAA was present on one allele of the PRNP gene only, so E211K mutation in the Canadian BSE 20 was heterozygous in nature. The finding of the E211K mutation was of significant interest because an analogous mutation, E200K, at codon 200 in human PRNP gene is associated with genetic CJD in humans [Citation28].

Figure 4. DNA sequencing results of the PRNP gene of the Canadian BSE case 20. DNA sequence chromatograms of the PRNP gene from A) brain stem, B) retropharyngeal lymph node, C) quadriceps, and D) tongue of the Canadian BSE case 20. The site of mutation is highlighted by a boxed area with a black arrow. Encoded amino acid sequence is mentioned at the bottom of each chromatogram by capital letters indicating a change of amino acid code from GAA to AAA resulting in a switch of amino acid from glutamic acid (E) to lysine (K). DNA sequence chromatograms are shown for forward (top) and reverse (bottom) primers for each tissue tested.

Figure 4. DNA sequencing results of the PRNP gene of the Canadian BSE case 20. DNA sequence chromatograms of the PRNP gene from A) brain stem, B) retropharyngeal lymph node, C) quadriceps, and D) tongue of the Canadian BSE case 20. The site of mutation is highlighted by a boxed area with a black arrow. Encoded amino acid sequence is mentioned at the bottom of each chromatogram by capital letters indicating a change of amino acid code from GAA to AAA resulting in a switch of amino acid from glutamic acid (E) to lysine (K). DNA sequence chromatograms are shown for forward (top) and reverse (bottom) primers for each tissue tested. Display full size The change in the amino acid sequence at codon 211 of the bovine PRNP gene has been shown to increase the susceptibility of cattle to H-BSE making it a pathogenic mutation in cattle. The E211K mutation was previously reported in an American H-BSE case in 2006 [Citation18], which was later found to be genetically inherited to its offspring (a heifer) [Citation21] indicating that the US E211K mutation is a germ line mutation. To determine if the E211K mutation identified in the Canadian BSE case 20 was a somatic or germline in nature, additional tissues were analysed. The DNA sequencing results on retropharyngeal lymph node, tongue and quadriceps also showed a change in the nucleotide sequence at codon 211 resulting in the E211K mutation (Figure 4). These results strongly suggest that E211K in the Canadian BSE case 20 was present as a germline mutation. Due to the poor condition of the carcass at post-mortem sample collection, neither blood nor gonadal tissues were available for further analyses.

Discussion

Here we describe a case of H-BSE confirmed in December 2021 in Canada (referred to as the Canadian BSE case 20). This was an 8-year-old dairy cow of Red Poll breed, born and raised in the C-BSE affected region in Canada. However, it was born after the reinforced feed ban (BARB) on feeding meat and bone meal to ruminants which supported the notion that this was not a C-BSE case. The laboratory confirmatory tests also verified that this was an H-BSE case based on the higher molecular weight of unglycosylated PrP in comparison to C-BSE and L-BSE controls, both before and after PNGase F treatment. Further analyses of glycoform ratios and the presence of a fourth band in the Canadian BSE case 20 were also consistent with the H-BSE control, as well as previously reported cases of H-BSE (Table 4) in the literature [Citation26]. Immunoreactivity with an N-terminal anti-PrP P4 antibody and increased sensitivity to stringent PK digestion conditions also confirmed this to be an H-BSE case (Table 4). These results were further supported by the presence of intra-neuronal, intra-microglial, and absence of linear and stellate PrPSc immunostaining pattern, which are characteristic features of H-BSE cases [Citation29]. Immediately, after the confirmation and typing of the Canadian BSE case 20, a notification was sent to WOAH and all the activities required for compliance with WOAH standards were conducted to ensure that the carcass was secured on the farm from the time of euthanasia until laboratory confirmation of BSE status and type. The carcass was then transported to the Canadian BSE RL for necropsy and incineration following post-mortem sample collection. During these activities, no part of the carcass of the Canadian BSE case 20 was rendered or entered the human food nor animal feed chain.

Table 4. Summary of comparison of characteristics of Canadian BSE cases to date.

Download CSV Display Table Speculating about the distribution of PrPSc in the peripheral tissues of any atypical BSE case is difficult because published data on the tissue tropism of PrPSc in cattle affected by atypical BSE are limited. However, this information is important for developing a good understanding of the evolution in prion strain biology and disease phenotype in newly identified BSE cases, particularly for atypical BSE cases. According to experimental transmission studies of atypical BSE, PrPSc has been reported in peripheral ganglia and nerves, muscles (muscle spindles), adrenal glands and retina for both H-BSE and L-BSE. No lymphoid tissues or gastrointestinal tissues have previously tested positive in atypical BSE cases [Citation30–34]. In the Canadian BSE case 20, the peripheral nervous system (PNS) including optic nerve was found to be 8.8% positive (relative to the obex) in IDEXX-EIA whereas the trigeminal ganglion and vagus nerve showed granular coarse PrPSc staining only in occasional neurons, as confirmed with two anti-PrP antibodies. However, brachial nerve plexus and sciatic nerve, which have been previously shown to be positive for PrPSc during experimental challenge studies and in natural cases [Citation35], were found to be negative in both IDEXX-EIA and IHC in the Canadian BSE case 20. Additionally, all lymphoid tissues as well as endo/exocrine glands examined were also negative in both IDEXX-EIA and IHC. Furthermore, striated muscles (including tongue and masseter muscles), nasal and buccal cavities, conjunctiva and ileocaecal junction also appeared negative in both tests. Amongst the muscles tested, the quadriceps and biceps brachii were negative in IDEXX-EIA; but showed inconclusive results in IHC. Overall, these results indicate that the distribution of the pathological agent is mainly restricted to the CNS with no or occasional weak involvement of the peripheral tissues in the Canadian BSE case 20. This also suggests the unlikely involvement of any new or unusual tissues during the disease pathogenesis of this Canadian BSE case 20, indicating no concerns that would require the revision of existing mitigation strategies nor the list of specified risk materials.

Regarding the origin of H-BSE cases, most of those reported worldwide have been sporadic except for one in the 2006 US H-BSE case associated with the E211K mutation [Citation18]. Canada has reported a total of 20 cases of BSE to date which included 17 C-BSE, 1 L-BSE, and 2 H-BSE cases. The only L-BSE case was identified in December 2007. The first case of H-BSE was identified in July 2006 and the latest case in December 2021.

To determine the involvement of host genetic risk factors, every field case of BSE is sequenced to analyse mutations in the PRNP gene. None of the previously reported pathogenic variations such as, i)-E211K mutation, ii)-23-bp indel in the promoter region, iii)-12-bp indel in intron 1, or iv)-increase in the number of OPRs on the N-terminal [Citation17–19] of the PRNP gene that are linked with enhanced susceptibility to BSE have been found in the first 19 BSE cases (including the first two atypical BSE cases) in Canada. However, synonymous mutations at codon 78 (Q78Q; CAG/CAA), 113 (P113P; CCC/CCT) and 192 (N192N; AAC/AAT), which have no known association with BSE, have been previously identified (Table 4) in some Canadian BSE cases [Citation23].

The DNA sequencing results of the Canadian BSE case 20 also showed no variation in the number of OPRs of the PRNP gene just like the previous Canadian BSE cases indicating a very low risk of such genetic variations in the Canadian cattle herd. Similarly, none of the synonymous mutations at codon 23 (L23L; CTC/CTA or CTG), 78 (Q78Q; CAG/CAA), 113 (P113P; CCC/CCT) and 185 (N185N; AAC/AAT), which were reported previously in the French, American, Swedish, Canadian and Portuguese H-BSE cases [Citation23,Citation35–38], were identified in the Canadian BSE case 20. Though a synonymous mutation at codon 192 (N192N; AAC/AAT) was detected in the Canadian BSE case 20, it was not a major concern because this mutation was also described previously in above mentioned studies and has no association with BSE. However, the detection of the E211K mutation in the Canadian BSE case 20 raises concerns because the E211K mutation has been associated with an increased susceptibility to H-BSE [Citation22] and is genetically inheritable to its offspring [Citation21], indicating that the E211K mutation could result in additional BSE cases.

This is the first time that the E211K mutation has been found in a Canadian BSE case. Additionally, the detection of the E211K mutation in multiple tissues strongly suggested that the E211K mutation in the Canadian BSE case 20 is a germline mutation. This implies that the E211K mutation in the Canadian BSE case 20 is most likely heritable and may be present in 50% of the offspring of this cow. However, it is difficult to speculate from where the Canadian BSE case 20 acquired the E211K mutation. The mutation could be either spontaneous or inherited from a parent. Possible theories about the origin of this E211K mutation in the Canadian BSE case 20 could include that, i) this cow acquired E211K mutation spontaneously in its early embryonic development, ii) germ cells from one of its parents acquired this mutation spontaneously, iii) ancestors of this cow had this mutation in their line, or iv) this mutation was present in a population of this breed. To explore these possibilities, a strategic testing of various cattle breeds (including Red Poll) from different geographic regions within Canada is important and being planned. However, none of its ancestors were traceable or available for testing to explore if the Canadian BSE case 20 acquired the E211K mutation from its ancestors. On the other hand, irrespective of the detection of the E211K mutation in this Canadian cow, there will always be a slight possibility for the E211K mutation to occur independently in any cattle. Nonetheless, the presence of the E211K mutation in cattle makes them highly susceptible to develop H-BSE [Citation22]. While strong BSE mitigation measures such as specified risk material removal, and ruminant to ruminant feed bans play a major role in preventing future BSE cases, the potential for animals with the E211K mutation and an increased risk of developing atypical BSE in the Canadian cattle population cannot be ruled out.

The E211K mutation associated with the US H-BSE case was heritable [Citation21] and associated with an increased susceptibility to H-BSE, represented by a shorter incubation time in cattle with K211 allele during the experimental challenge studies [Citation22,Citation39], and suggested a higher propensity for the K211 prion protein to misfold. Identification of E211K mutation in the 2006 US and 2021 Canadian H-BSE cases represent independent existence of this pathogenic mutation in time and space. This is further supported by the comparative analysis (Supplementary Table S1) of both cases showing minor dissimilarities, though both cases were confirmed as H-type BSE. Those dissimilarities included the presence of 4th band of PK resistant PrP at 10 kDa in western-blot and a synonymous mutation at codon 192 of the PRNP gene in Canadian BSE cases 20 only. Additionally, proven heritability and germline nature of E211K mutation in the 2006 US and 2021 Canadian H-BSE cases respectively, indicate familial nature of E211K mutation just like familial Creutzfeldt Jakob’s disease in humans. This highlights the importance of screening each identified case for genetic risk factors specifically and cattle population to determine the prevalence rate of E211K mutation in general, to eradicate this pathogenic mutation form the national herd.

After the detection of E211K mutation in the US H-BSE case in 2006, DNA testing of cattle populations was conducted in various countries including USA (n = 6062 cattle tested) [Citation40], China (n = 349) [Citation41], Poland (n = 105) [Citation42], Pakistan (n = 236 cattle and n = 281 buffalo) [Citation43] and Korea (n = 536) [Citation44], but no E211K mutation could be detected in any sample. This indicates that the E211K mutation in cattle is very rare. Notably, all the previously mentioned studies have largely analysed blood samples with only occasional testing of muscle or brain tissues to detect the E211K mutation, which might not be detected if it was present as a somatic or postzygotic mutation within a particular tissue. Recently, Won SY and colleagues novelly reported that the E211K mutation was present only in the medulla oblongata of three Holstein Korean slaughter cattle, was present at high rates and was a somatic mutation. In silico analyses of the biological impact of the E211K mutation in these cattle predicted that E211K was damaging in nature despite being a somatic mutation [Citation45]. Somatic mutations in the PRNP gene at codon D178N in sporadic CJD and P102L associated with sporadic Gerstmann – Sträussler – Scheinker (GSS) syndrome have also been documented in humans [Citation46,Citation47]. This indicates that the E211K mutation can be pathogenic irrespective of whether it is germline or somatic in origin. However, PrPSc was not detected in the medulla oblongata of the three Holstein Korean cattle carrying E211K as a somatic mutation, using highly sensitive in-vitro amplification assays including real-time quaking-induced conversion reactions (RT-QuIC) and protein misfolding cyclic amplification (PMCA) [Citation48]. Though more objective evidence is required to explore the possibility that a somatic E211K mutation can cause H-BSE, results from Kim YC and colleagues show that the level of somatic mutation required to induce H-BSE could be a determining factor. On the other hand, it could be argued that the three Holstein Korean cattle carrying a somatic E211K mutation had an average age of less than 5 years, and they might have acquired atypical BSE if they were let to live longer. While scientific evidence exist that E211K mutation increases susceptibility to H-BSE, however it is difficult to objectively conclude that E211K mutation is a bona fide mutation responsible for the onset of H- BSE. In this regard, a long-term project is already underway by the USDA to investigate the possibility of a genetic aetiology for H-BSE in cattle expressing the K211 allele. Based on the results of this study, and the 2006 H-BSE case in the USA, there is an expanded spectrum of aetiologies for bovine prion diseases similar to what is observed in humans, including sporadic, genetic and acquired versions.

Supplemental material Canadian 2021 H-type Bovine Spongiform Encephalopathy case associated with a novel E211K polymorphism in prion protein gene

KEYWORDS:

Atypical BSEBovine Spongiform Encephalopathycentral nervous systemE211K mutationprion diseasesprion protein genesynonymous mutation

https://www.tandfonline.com/doi/full/10.1080/19336896.2025.2511933#d1e1606

“Based on the results of this study, and the 2006 H-BSE case in the USA, there is an expanded spectrum of aetiologies for bovine prion diseases similar to what is observed in humans, including sporadic, genetic and acquired versions.”

https://www.tandfonline.com/doi/full/10.1080/19336896.2025.2511933#d1e1606

See Canadian 2021 Mad Cow

SATURDAY, DECEMBER 18, 2021

CFIA Canada Alberta Laboratory detection of atypical bovine spongiform encephalopathy

https://bovineprp.blogspot.com/2021/12/cfia-canada-alberta-laboratory.html

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 ??? let’s analyze the science on this tse prion E200K, shall we…

Cattle with the E211K polymorphism, and gCJD linked to a glutamic acid to lysine substitution at codon 200 (E200K) of PRNP, what if?

i believe science has shown that to many times with Transmissible Spongiform Encephalopathy TSE PrP disease, and all of the variants, strains, too many times scientists are to quick ruling out potential causes or links, before actually proving their findings. i believe that science shows that feed can potentially play a role in Cattle with the E211K polymorphism, and there from, humans with gCJD linked to a glutamic acid to lysine substitution at codon 200 (E200K) of PRNP, and or iatrogenic events, there from, what if?

AI overview

The E211K mutation in the bovine PRNP gene is a critical, rare, and heritable genetic variation associated with H-type bovine spongiform encephalopathy (H-BSE). It is considered to be the bovine equivalent to the human PRNP E200K mutation, which causes the most common form of genetic Creutzfeldt-Jakob disease (gCJD).

Genetic Creutzfeldt-Jakob disease linked to the E200K mutation: a large cohort study Published: 13 January 2026

Abstract

Creutzfeldt-Jakob disease (CJD), the most common human prion disease, is an invariably fatal neurodegenerative disorder affecting 1.5 cases per million individuals per year. About 10–15% of the human prion diseases are caused by a pathogenic variant in the prion protein (PrP) gene (PRNP), and the most common genetic human prion disease is CJD (gCJD) linked to a glutamic acid to lysine substitution at codon 200 (E200K) of PRNP. The polymorphic codon 129 methionine (M)/valine (V) genotype has a strong effect on disease phenotype. In the present study, we retrospectively evaluated many features of gCJD E200K cases with respect to the 129MV polymorphism, type of scrapie prion protein (PrPSc), demographic, clinical, laboratory, histopathology, and molecular features, including western blot examination and real-time quaking-induced conversion assay. Analyses were also performed to determine statistically significant features between E200K haplotypes (e.g., codon 129 genotype in cis with the mutated allele) and codon 129 genotypes. This study found that codon 129 polymorphism affects several disease features of gCJD E200K. Specifically, histopathologic differences were found between patients with different 129 haplotypes and genotypes. We have identified five groups or subtypes of E200K associated with either PrPSctype 1 or 2. Other E200K cases showed mixed (i) PrPSc types or (ii) pathological features of 129 M and 129 V haplotypes. To our knowledge, this study describes the largest cohort of 177 E200K cases and provides new insight into the wide range of phenotypes associated with this common CJD genetic variant.

https://link.springer.com/article/10.1007/s00401-026-02975-x

Conclusions: Cattle with the E211K polymorphism are susceptible to the CWD agent after oronasal exposure of 0.2 g of infectious material.

Funded by: This research was funded in its entirety by congressionally appropriated funds to the United States Department of Agriculture, Agricultural Research Service. The funders of the work did not influence study design, data collection and analysis, decision to publish, or preparation of the manuscript.

"Cattle with the E211K polymorphism are susceptible to the CWD agent after oronasal exposure of 0.2 g of infectious material."

https://prion2023.org/wp-content/uploads/2023/10/Meeting-book-final-version2.pdf

https://web.archive.org/web/20250828201533/https://prion2023.org/wp-content/uploads/2023/10/Meeting-book-final-version2.pdf

https://www.researchgate.net/profile/Syed-Zahid-Shah/publication/378314391_Meeting-book-final-version_prion_2023/links/65d44dad28b7720cecdca95f/Meeting-book-final-version-prion-2023.pdf

Title: Short incubation periods of atypical H-type BSE to cattle with EK211 and KK211 prion protein genotypes after intracranial inoculation

https://www.ars.usda.gov/research/publications/publication/?seqNo115=408989#:~:text=Technical%20Abstract:%20In%202006%20a,PrPSc%20immunohistochemistry%2C%20and%20western%20blot.

First Report of the Potential Bovine Spongiform Encephalopathy (BSE)-Related Somatic Mutation E211K of the Prion Protein Gene (PRNP) in Cattle

Sae-Young Won 1,2,† , Yong-Chan Kim 1,2,† and Byung-Hoon Jeong 1,2,*

1 Korea Zoonosis Research Institute, Jeonbuk National University, Iksan, Jeonbuk 54531, Korea; gkfh32@jbnu.ac.kr (S.-Y.W.); kych@jbnu.ac.kr (Y.-C.K.) 2 Department of Bioactive Material Sciences and Institute for Molecular Biology and Genetics, Jeonbuk National University, Jeonju, Jeonbuk 54896, Korea * Correspondence: bhjeong@jbnu.ac.kr; Tel.: +82-63-900-4040; Fax: +82-63-900-4012 † These authors contributed equally to this work.

Received: 25 May 2020; Accepted: 11 June 2020; Published: 15 June 2020

Abstract: Bovine spongiform encephalopathy (BSE) is a prion disease characterized by spongiform degeneration and astrocytosis in the brain. Unlike classical BSE, which is caused by prion-diseasecontaminated meat and bone meal, the cause of atypical BSE has not been determined. Since previous studies have reported that the somatic mutation in the human prion protein gene (PRNP) has been linked to human prion disease, the somatic mutation of the PRNP gene was presumed to be one cause of prion disease. However, to the best of our knowledge, the somatic mutation of this gene in cattle has not been investigated to date. We investigated somatic mutations in a total of 58 samples, including peripheral blood; brain tissue including the medulla oblongata, cerebellum, cortex, and thalamus; and skin tissue in 20 individuals from each breed using pyrosequencing. In addition, we estimated the deleterious effect of the K211 somatic mutation on bovine prion protein by in silico evaluation tools, including PolyPhen-2 and PANTHER. We found a high rate of K211 somatic mutations of the bovine PRNP gene in the medulla oblongata of three Holsteins (10% ± 4.4%, 28% ± 2%, and 19.55% ± 3.1%). In addition, in silico programs showed that the K211 somatic mutation was damaging. To the best of our knowledge, this study is the first to investigate K211 somatic mutations of the bovine PRNP gene that are associated with potential BSE progression.

snip...

3. Discussion

Although the number of classical BSE cases caused by contaminated meat and bone meal has decreased dramatically due to global efforts, the number of atypical BSE cases increased [30,31]. In humans, genetic prion disease makes up approximately 10% to 15% of cases with germline mutations of the PRNP gene [19]. However, although sporadic prion disease accounts for 85% of human prion diseases, the exact cause of sporadic prion disease has not been revealed to date. In recent studies, somatic mutations in the PRNP gene have been identified in human sporadic prion diseases and have been suggested as one cause of sporadic prion disease. In cattle, the E211K germline mutation of the bovine PRNP gene was first reported in the United States in 2006 [25–27]. The bovine E211K mutation showed homology in the region with the E200K mutation of the human PRNP gene, which is most frequently observed in human familial prion diseases [20,22]. Thus, we investigated the E211K somatic mutation of the bovine PRNP gene, which may be considered a novel risk factor for BSE in Korean cattle.

In our previous study, the germline mutation at codon 211 of the bovine PRNP gene was not observed in 384 Hanwoo cattle and 152 Holstein cattle [5]. Remarkably, we found high rates of K211 somatic mutation of the bovine PRNP gene in three Holstein cattle (Table 3). In Korea, since classical and atypical BSEs have never been reported in Hanwoo, the absence of K211 somatic mutations in Hanwoo is notable. In addition, although we tested K211 somatic mutations, including whole blood and four brain regions, somatic mutations of this gene were detected only in the medulla oblongata. The expected level of K211 mutations of the bovine PRNP gene, which can initiate atypical BSE, is elusive. A previous study in early-onset Alzheimer’s disease reported that 14% of mutations of the presenilin 1 (PSEN1) gene in brain cells are responsible for the initiation of this disease [32]. In another study, the somatic mutations of amyloid precursor protein (APP), nicastrin (NCSTN), sortilin-related receptor (SORL1), and microtubule affinity-regulating kinase 4 (MARK4) were observed in sporadic Alzheimer’s disease patients in 0.2–10.5% [33]. We observed three samples containing over 10% K211 mutations (Figure 3). Thus, BSE inspection of these samples seems essential to elucidate the expected level of K211 mutations of the PRNP gene, which can initiate atypical BSE in the future. According to previous studies, since the medulla oblongata showed a prominent accumulation of PrPSc, the diagnosis of BSE was performed in the medulla oblongata region [34]. However, since the three Holstein cattle carrying the K211 somatic mutation in the medulla oblongata were not investigated as to whether K211 somatic mutation could be observed in other brain regions, it is difficult to conclude that this somatic mutation was only found in the medulla oblongata. In addition, since atypical BSE has shown to be a prominent brain pathology in the frontal cortex [17,18], if the atypical BSE was caused by a somatic mutation of the PRNP gene, it is expected that the somatic mutation was also found in the cerebral cortex. Thus, further investigation of the K211 somatic mutation is highly desirable in other brain regions, including the cerebral cortex of the three Holstein cattle carrying K211 somatic mutation in the medulla oblongata. We also performed in silico analysis using PolyPhen-2 and PANTHER to evaluate whether the mutation of K211 of the bovine PRNP gene affects the bovine PrP protein. Notably, the K211 somatic mutation of the bovine PRNP gene was evaluated as “damaging”. Since PolyPhen-2 can estimate structural variation, the prediction can be interpreted that K211 somatic mutation can contribute to the conformational change to the susceptible structure of the PrPSc. In addition, the wild-type PrP with E211 allele was predicted to be of high preservation time (361, Table 4) and K211 somatic mutation was estimated to be deleterious by using the PANTHER program. The prediction of PANTHER indicated that K211 mutation has been very rare and has not been observed in ancestral proteins for a long time. To confirm the effect of K211 mutation on prion disease, investigation of symptoms of prion disease in bovine PrP transgenic mice carrying the K211 allele is highly desirable in the future. It seems possible that a high somatic mutation rate contributes to sporadic prion disease in cattle.

Previous studies have reported that prion disease is accelerated in transgenic (Tg) mice expressing wild-type bank vole prion protein (BVPrP) containing E200K, which is associated with human familial CJD [35]. In addition, recent studies have demonstrated that a human PrP Tg mouse model with the E200K mutation can lead to spontaneous prion disease [36]. In cattle, the E211K mutation, which is a homologous region of human E200K, is expected to have a deleterious effect on BSE, and several in silico programs in the present study are expected to cause structural changes in the bovine prion protein (Table 4). To determine whether the E211K mutation affects the susceptibility to prion disease, brain samples with high mutation rates of E211K should be investigated by protein misfolding cyclic amplification (PMCA) or Western blotting (WB) in the future.

https://www.mdpi.com/1422-0067/21/12/4246/pdf

Canadian 2021 H-type Bovine Spongiform Encephalopathy case associated with a novel E211K polymorphism in prion protein gene Waqas Tahir , Sandor Dudas , Renee Anderson , Jianmin Yang , Sarah Bogart , Kristina Santiago-Mateo , show all Pages 36-49 | Received 20 Feb 2025, Accepted 22 May 2025, Published online: 04 Aug 2025 Cite this article

https://doi.org/10.1080/19336896.2025.2511933

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ABSTRACT Bovine Spongiform Encephalopathy (BSE) is a fatal neurodegenerative disease in cattle which can be either classical BSE (C-BSE) or atypical BSE (including H-BSE and L-BSE). Here, we report the results of our analyses of an H-BSE case found in Canada in 2021, indicating restriction of the pathological agent (PrPSc) mainly to the central nervous system with no or occasional weak involvement of peripheral tissues. Importantly, a non-synonymous mutation at codon 211 of the PRNP gene was detected and confirmed to be present as a germline mutation. This is the first case of BSE in Canada with a predisposing E211K mutation.

snip…

Somatic mutations in the PRNP gene at codon D178N in sporadic CJD and P102L associated with sporadic Gerstmann – Sträussler – Scheinker (GSS) syndrome have also been documented in humans 46, 47.

https://www.tandfonline.com/doi/full/10.1080/19336896.2025.2511933#d1e1606

Mad Cow Disease BSE, could it be here 2001 revisited 2026

https://bovineprp.blogspot.com/2026/08/mad-cow-disease-could-it-be-here-2001.html

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

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 argely 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.

http://web.archive.org/web/20140912011905/http://www.prion2009.com/sites/default/files/Prion2009_Book_of_Abstracts.pdf

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 argely 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.

http://www.neuroprion.org/resources/pdf_docs/conferences/prion2009/prion2009_bookofabstracts.pdf

see full text ;

http://bse-atypical.blogspot.com/2009/10/atypical-bse-bse-and-other-human-and.html

snip...

full text Singeltary et al PLOS

http://journals.plos.org/plosone/article/comment?id=info:doi/10.1371/annotation/4f9be886-69fe-4c7c-922b-85b0ecbe6d53

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.ukJürgen A. Richt College of Veterinary Medicine, Kansas State University, K224B Mosier Hall, Manhattan, Kansas 66506-5601, USA NATURE|Vol 457|26 February 2009

http://www.nature.com/nature/journal/v457/n7233/full/4571079b.html

Title: Transmission of atypical BSE: a possible origin of Classical BSE in cattle

Authors: Sandor Dudas1, Samuel James Sharpe1, Kristina Santiago-Mateo1, Stefanie Czub1, Waqas Tahir1,2, *

Affiliation: 1National and WOAH reference Laboratory for Bovine Spongiform Encephalopathy, Canadian Food inspection Agency, Lethbridge Laboratory, Lethbridge, Canada. 2Department of Biological Sciences, University of Lethbridge, Lethbridge, Alberta, Canada.

*Corresponding and Presenting Author: waqas.tahir@inspection.gc.ca

Background: Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative disease of cattle and is categorized into classical and atypical forms. Classical BSE (CBSE) is linked to the consumption of BSE contaminated feed whereas atypical BSE is considered to be spontaneous in origin. The potential for oral transmission of atypical BSE is yet to be clearly defined.

Aims: To assess the oral transmissibility of atypical BSE (H and L type) in cattle. Should transmission be successful, determine the biochemical characteristics and distribution of PrPSc in the challenge cattle.

Material and Methods: For oral transmission, calves were fed with 100 g of either H (n=3) or L BSE (n=3) positive brain material. Two years post challenge, 1 calf from each of the H and L BSE challenge groups exhibited behavioural signs and were euthanized. Various brain regions of both animals were tested by traditional and novel prion detection methods with inconclusive results. To detect infectivity, brain homogenates from these oral challenge animals (P1) were injected intra-cranially (IC) into steer calves. Upon clinical signs of BSE, 3/4 of IC challenged steer calves were euthanized and tested for PrPSc with ELISA, immunohistochemistry and immunoblot.

Results: After 6 years of incubation, 3/4 animals (2/2 steers IC challenged with brain from P1 L-BSE oral challenge and 1/2 steer IC challenged with brain from P1 H-BSE oral challenge) developed clinical disease. Analysis of these animals revealed high levels of PrPSc in their brains, having biochemical properties similar to that of PrPSc in C-BSE.

Conclusion: These results demonstrate the oral transmission potential of atypical BSE in cattle. Surprisingly, regardless of which atypical type of BSE was used for P1 oral challenge, PrPSc in the P2 animals acquired biochemical characteristics similar to that of PrPSc in C-BSE, suggesting atypical BSE as a possible origin of C-BSE in UK.

Presentation Type: Oral Presentation

Funded by: CFIA, Health Canada, Alberta Livestock and Meat Agency, Alberta Prion Research Institute

Grant Number: ALMA/APRI: 201400006, HC 414250

https://prion2023.org/wp-content/uploads/2023/10/Meeting-book-final-version2.pdf

https://web.archive.org/web/20250828201533/https://prion2023.org/wp-content/uploads/2023/10/Meeting-book-final-version2.pdf

https://www.researchgate.net/profile/Syed-Zahid-Shah/publication/378314391_Meeting-book-final-version_prion_2023/links/65d44dad28b7720cecdca95f/Meeting-book-final-version-prion-2023.pdf

OIE Conclusions on transmissibility of atypical BSE among cattle

Given that cattle have been successfully infected by the oral route, at least for L-BSE, it is reasonable to conclude that atypical BSE is potentially capable of being recycled in a cattle population if cattle are exposed to contaminated feed. In addition, based on reports of atypical BSE from several countries that have not had C-BSE, it appears likely that atypical BSE would arise as a spontaneous disease in any country, albeit at a very low incidence in old cattle. In the presence of livestock industry practices that would allow it to be recycled in the cattle feed chain, it is likely that some level of exposure and transmission may occur. As a result, since atypical BSE can be reasonably considered to pose a potential background level of risk for any country with cattle, the recycling of both classical and atypical strains in the cattle and broader ruminant populations should be avoided.

https://www.oie.int/fileadmin/SST/adhocreports/Bovine%20spongiform%20encephalopathy/AN/A_AhG_BSEsurv_RiskAss_Mar2019.pdf

Annex 7 (contd) AHG on BSE risk assessment and surveillance/March 2019

34 Scientific Commission/September 2019

3. Atypical BSE

The Group discussed and endorsed with minor revisions an overview of relevant literature on the risk of atypical BSE being recycled in a cattle population and its zoonotic potential that had been prepared ahead of the meeting by one expert from the Group. This overview is provided as Appendix IV and its main conclusions are outlined below. With regard to the risk of recycling of atypical BSE, recently published research confirmed that the L-type BSE prion (a type of atypical BSE prion) may be orally transmitted to calves1 . In light of this evidence, and the likelihood that atypical BSE could arise as a spontaneous disease in any country, albeit at a very low incidence, the Group was of the opinion that it would be reasonable to conclude that atypical BSE is potentially capable of being recycled in a cattle population if cattle were to be exposed to contaminated feed. Therefore, the recycling of atypical strains in cattle and broader ruminant populations should be avoided.

4. Definitions of meat-and-bone meal (MBM) and greaves

http://web.oie.int/downld/PROC2020/A_SCAD_Sept2019.pdf

The L-type BSE prion is much more virulent in primates and in humanized mice than is the classical BSE prion, which suggests the possibility of zoonotic risk associated with the L-type BSE prion

https://wwwnc.cdc.gov/eid/article/16/7/09-1882_article

Consumption of L-BSE–contaminated feed may pose a risk for oral transmission of the disease agent to cattle.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5324790/

Thus, it is imperative to maintain measures that prevent the entry of tissues from cattle possibly infected with the agent of L-BSE into the food chain.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3310119/

Atypical L-type bovine spongiform encephalopathy (L-BSE) transmission to cynomolgus macaques, a non-human primate

Fumiko Ono 1, Naomi Tase, Asuka Kurosawa, Akio Hiyaoka, Atsushi Ohyama, Yukio Tezuka, Naomi Wada, Yuko Sato, Minoru Tobiume, Ken'ichi Hagiwara, Yoshio Yamakawa, Keiji Terao, Tetsutaro Sata

Affiliations expand

PMID: 21266763

Abstract

A low molecular weight type of atypical bovine spongiform encephalopathy (L-BSE) was transmitted to two cynomolgus macaques by intracerebral inoculation of a brain homogenate of cattle with atypical BSE detected in Japan. They developed neurological signs and symptoms at 19 or 20 months post-inoculation and were euthanized 6 months after the onset of total paralysis. Both the incubation period and duration of the disease were shorter than those for experimental transmission of classical BSE (C-BSE) into macaques. Although the clinical manifestations, such as tremor, myoclonic jerking, and paralysis, were similar to those induced upon C-BSE transmission, no premonitory symptoms, such as hyperekplexia and depression, were evident. Most of the abnormal prion protein (PrP(Sc)) was confined to the tissues of the central nervous system, as determined by immunohistochemistry and Western blotting. The PrP(Sc) glycoform that accumulated in the monkey brain showed a similar profile to that of L-BSE and consistent with that in the cattle brain used as the inoculant. PrP(Sc) staining in the cerebral cortex showed a diffuse synaptic pattern by immunohistochemistry, whereas it accumulated as fine and coarse granules and/or small plaques in the cerebellar cortex and brain stem. Severe spongiosis spread widely in the cerebral cortex, whereas florid plaques, a hallmark of variant Creutzfeldt-Jakob disease in humans, were observed in macaques inoculated with C-BSE but not in those inoculated with L-BSE.

https://pubmed.ncbi.nlm.nih.gov/21266763/

see full text;

https://www.niid.go.jp/niid/images/JJID/64/81.pdf

***> Atypical BSE in cattle

THE recent diagnosis of two atypical bovine spongiform encephalopathy (BSE) cases in Great Britain (March 2023 in Cornwall and December 2024 in Dumfries and Galloway) and one in the Republic of Ireland (in November 2023) warrants a reminder about this notifiable disease.

Since 2005, a total of 17 cases have been detected in Great Britain.1 Unlike classical BSE, which resulted in over 180,000 cases in Great Britain and was predominantly associated with the consumption of feed contaminated with the BSE agent, and where the last case was confirmed in Ayrshire in May 2024, atypical BSE is believed to be a spontaneous disease in cattle found in approximately one in 1,000,000 tested cattle based on French data,2 similar to the sporadic Creutzfeldt- Jakob disease in people. There is currently no evidence that atypical BSE causes a disease in people, although it can be transmitted experimentally to other species by intracerebral inoculation, including primates.3–5 The World Organisation for Animal Health does not include atypical BSE in its geographical BSE risk status assessment.

Despite differences in terms of epidemiological, molecular and biological phenotype compared with classical BSE, atypical BSE is currently treated as if it were classical BSE in accordance with EU and UK legislation: once a case is identified, all cohort animals born and reared with the affected animal during the first 12 months of its life, and all offspring born within 24 months of its clinical onset, are culled and tested for BSE, which does seem to be at odds with the hypothesis that it is a spontaneous disease. This is more a precautionary measure to maintain confidence in the beef trade and protect consumers while more knowledge about this disease is obtained.

Almost all current knowledge on atypical BSE is based on experimental infection because this spontaneous

VET RECORD | 29 March–12 April 2025

disease has generally only been found in aged downer cows, which is difficult to replicate experimentally in the host species. Intracerebral inoculation of brain tissue from an affected cow causes disease in cattle in less than two years, unlike the natural disease that usually occurs in animals over eight years of age.

The vast majority of cases have been identified by active monitoring of fallen stock or emergency slaughter of cattle, where only the brain sample of various stages of autolysis is generally available. Little is known of where the atypical BSE agent can be found in natural disease, other than in the brain, because all the cases confirmed have been identified after death through active surveillance, by which time most peripheral tissue has been disposed of. Limited material from a single case of a naturally affected cow was tested in Italy by mouse bioassay, which found infectivity in muscle.6 In experimental disease generated by intracerebral inoculation of cattle, infectivity can be detected in the brain and spinal cord, ganglia, peripheral nerves and skeletal muscles, similar to classical BSE, but not in peripheral lymphoid tissue.6–8

Early reporting of clinical suspects is needed so that the live animal or the whole carcase can be delivered to an APHA regional laboratory for tissue sampling. This is made more difficult due to the subtlety of clinical signs based on experimental disease. Clinical cases may not be as over- reactive or nervous as classical BSE cases; some may, in fact, be dull, but what most cases have in common is that they have difficulty getting up and eventually end up as downer cows, and only the clinical history may reveal some prior behavioural or locomotor changes. High creatinine kinase serum levels and nibbling in response to scratching the tail head or back were some features in experimental disease,8, 9 but it is not known whether this is also seen in natural disease.

In general, BSE should be considered as a differential diagnosis in all downer cows that do not respond to treatment, where the blood results do not support the presence of a metabolic disease and where the cause cannot be determined with confidence.

Since BSE is a notifiable disease, suspected cases of BSE in Great Britain must be reported to the local APHA office.

Changes are imminent in the reporting of fallen stock cattle, which will require the owner to state whether the animal displayed signs of changes in behaviour, sensation or locomotion before death, in addition to the likely cause of death or disease. This is to obtain a better profile of the clinical history, if cattle are retrospectively diagnosed as BSE cases, which has happened in all BSE cases confirmed since 2010: none has been reported as a clinical suspect.

“BSE should be considered as a differential diagnosis in all downer cows that do not respond to treatment”

Timm Konold, TSE lead scientist

Brenda Rajanayagam, workgroup leader for the data systems group

APHA Weybridge, New Haw, Addlestone, Surrey KT15 3NB email: timm.konold@apha.gov.uk

Keith Meldrum, former chief veterinary officer The Orchard, Swaynes Lane, Guildford, Surrey GU1 2XX

References

1 APHA. Cattle: TSE surveillance statistics. Overview of Great Britain statistics. 2025. https://bit.ly/4ho5Nds (accessed 19 March 2025)

Atypical BSE In Cattle

https://bvajournals.onlinelibrary.wiley.com/doi/10.1002/vetr.5400

***> Alabama Mad Cow Feed animal feeds for ruminants rule, 21 CFR 589.200 IN COMMERCE <***

10 years post mad cow feed ban August 1997

RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINE -- CLASS II

______________________________

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

______________________________

PRODUCT

Bulk custom dairy feds manufactured from concentrates, Recall # V-113-6

CODE

All dairy feeds produced between 2/1/05 and 6/16/06 and containing H. J. Baker recalled feed products.

RECALLING FIRM/MANUFACTURER

Vita Plus Corp., Gagetown, MI, by visit beginning on June 21, 2006. Firm initiated recall is complete.

REASON

The feed was manufactured from materials that may have been contaminated with mammalian protein.

VOLUME OF PRODUCT IN COMMERCE

27,694,240 lbs

DISTRIBUTION

MI

______________________________

PRODUCT

Bulk custom made dairy feed, Recall # V-114-6

CODE

None

RECALLING FIRM/MANUFACTURER

Burkmann Feeds LLC, Glasgow, KY, by letter on July 14, 2006. Firm initiated recall is ongoing.

REASON

Custom made feeds contain ingredient called Pro-Lak, which may contain ruminant derived meat and bone meal.

VOLUME OF PRODUCT IN COMMERCE

?????

DISTRIBUTION

KY

END OF ENFORCEMENT REPORT FOR AUGUST 2, 2006

###

http://data.nber.org/fda/enforcement-report/2006/ucm120413.htm

=====

PRODUCT

Bulk Whole Barley, Recall # V-256-2009

CODE

No code or lot number.

RECALLING FIRM/MANUFACTURER

Mars Petcare US, Clinton, OK, by telephone on May 21, 2009. Firm initiated recall is complete.

REASON

Product may have contained prohibited materials without cautionary statement on the label.

VOLUME OF PRODUCT IN COMMERCE

208,820 pounds

DISTRIBUTION

TX

END OF ENFORCEMENT REPORT FOR AUGUST 26, 2009

###

https://www.fda.gov/Safety/Recalls/EnforcementReports/ucm180348.htm

Subject: MAD COW FEED RECALL KY VOLUME OF PRODUCT IN COMMERCE ?????

Date: August 6, 2006 at 6:19 pm PST

PRODUCT Bulk custom made dairy feed, Recall # V-114-6

CODE None

RECALLING FIRM/MANUFACTURER Burkmann Feeds LLC, Glasgow, KY, by letter on July 14, 2006.

Firm initiated recall is ongoing. REASON Custom made feeds contain ingredient called Pro-Lak, which may contain ruminant derived meat and bone meal.

VOLUME OF PRODUCT IN COMMERCE ?????

DISTRIBUTION KY

END OF ENFORCEMENT REPORT FOR AUGUST 2, 2006

###

http://www.fda.gov/bbs/topics/enforce/2006/ENF00963.html

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

###

http://www.fda.gov/bbs/topics/enforce/2006/ENF00960.html

Subject: MAD COW FEED BAN WARNING LETTER ISSUED MAY 17, 2006

Date: June 27, 2006 at 7:42 am PST Public Health Service Food and Drug Administration

New Orleans District 297 Plus Park Blvd. Nashville, TN 37217

Telephone: 615-781-5380 Fax: 615-781-5391

May 17, 2006

WARNING LETTER NO. 2006-NOL-06

FEDERAL EXPRESS OVERNIGHT DELIVERY

Mr. William Shirley, Jr., Owner Louisiana.DBA Riegel By-Products 2621 State Street Dallas, Texas 75204

Dear Mr. Shirley:

On February 12, 17, 21, and 22, 2006, a U.S. Food & Drug Administration (FDA) investigator inspected your rendering plant, located at 509 Fortson Street, Shreveport, Louisiana. The inspection revealed significant deviations from the requirements set forth in Title 21, Code of Federal Regulations, Part 589.2000 [21 CFR 589.2000], Animal Proteins Prohibited in Ruminant Feed. This regulation is intended to prevent the establishment and amplification of Bovine Spongiform Encephalopathy (BSE). You failed to follow the requirements of this regulation; products being manufactured and distributed by your facility are misbranded within the meaning of Section 403(a)(1) [21 USC 343(a)(1)] of the Federal Food, Drug, and Cosmetic Act (the Act).

Our investigation found you failed to provide measures, including sufficient written procedures, to prevent commingling or cross-contamination and to maintain sufficient written procedures [21 CFR 589.2000(e)] because:

You failed to use clean-out procedures or other means adequate to prevent carryover of protein derived from mammalian tissues into animal protein or feeds which may be used for ruminants. For example, your facility uses the same equipment to process mammalian and poultry tissues. However, you use only hot water to clean the cookers between processing tissues from each species. You do not clean the auger, hammer mill, grinder, and spouts after processing mammalian tissues.

You failed to maintain written procedures specifying the clean-out procedures or other means to prevent carryover of protein derived from mammalian tissues into feeds which may be used for ruminants.

As a result . the poultry meal you manufacture may contain protein derived from mammalian tissues prohibited in ruminant feed. Pursuant to 21 CFR 589.2000(e)(1)(i), any products containing or may contain protein derived from mammalian tissues must be labeled, "Do not feed to cattle or other ruminants." Since you failed to label a product which may contain protein derived from mammalian tissues with the required cautionary statement. the poultry meal is misbranded under Section 403(a)(1) [21 USC 343(a)(1)] of the Act.

This letter is not intended as an all-inclusive list of violations at your facility. As a manufacturer of materials intended for animal feed use, you are responsible for ensuring your overall operation and the products you manufacture and distribute are in compliance with the law. You should take prompt action to correct these violations, and you should establish a system whereby violations do not recur. Failure to promptly correct these violations may result in regulatory action, such as seizure and/or injunction, without further notice.

You should notify this office in writing within 15 working days of receiving this letter, outlining the specific steps you have taken to bring your firm into compliance with the law. Your response should include an explanation of each step taken to correct the violations and prevent their recurrence. If corrective action cannot be completed within 15 working days, state the reason for the delay and the date by which the corrections will be completed. Include copies of any available documentation demonstrating corrections have been made.

Your reply should be directed to Mark W. Rivero, Compliance Officer, U.S. Food and Drug Administration, 2424 Edenborn Avenue, Suite 410, Metairie, Louisiana 70001. If you have questions regarding any issue in this letter, please contact Mr. Rivero at (504) 219-8818, extension 103.

Sincerely,

/S

Carol S. Sanchez Acting District Director New Orleans District

http://www.fda.gov/foi/warning_letters/g5883d.htm

PLEASE NOTE, THE FDA URLS FOR OLD WARNING LETTERS ARE OBSOLETE AND DO NOT WORK IN MOST CASES. I LOOKED UP THE OLD ONE ABOVE AND FOUND IT, BUT HAVE NOT DONE THAT FOR THE OTHERS TO FOLLOW. THE DATA IS VALID THOUGH!

Subject: MAD COW PROTEIN IN COMMERCE USA 2006 RECALL UPDATE

From: "Terry S. Singeltary Sr." <[log in to unmask]>

Reply-To: SAFETY <[log in to unmask]>

Date: Mon, 9 Oct 2006 14:10:37 -0500

Subject: MAD COW FEED RECALL USA SEPT 6, 2006 1961.72 TONS

IN COMMERCE AL, TN, AND WV

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

______________________________

snip...

http://www.fda.gov/bbs/topics/enforce/2006/ENF00968.html

Subject: MAD COW FEED RECALLS ENFORCEMENT REPORT FOR AUGUST 9, 2006 KY, LA, MS, AL, GA, AND TN 11,000+ TONS

Date: August 16, 2006 at 9:19 am PST RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINE - CLASS II

______________________________

snip...

______________________________

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

###

http://www.fda.gov/bbs/topics/ENFORCE/2006/ENF00964.html

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

###

http://www.fda.gov/bbs/topics/enforce/2006/ENF00963.html

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

###

http://www.fda.gov/bbs/topics/enforce/2006/ENF00960.html

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

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

RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINES -- CLASS II PRODUCT

Bulk cattle feed made with recalled Darling's 85% Blood Meal, Flash Dried, Recall # V-024-2007 CODE Cattle feed delivered between 01/12/2007 and 01/26/2007 RECALLING FIRM/MANUFACTURER Pfeiffer, Arno, Inc, Greenbush, WI. by conversation on February 5, 2007.

Firm initiated recall is ongoing.

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

VOLUME OF PRODUCT IN COMMERCE 42,090 lbs. DISTRIBUTION WI

___________________________________

PRODUCT

Custom dairy premix products:

MNM ALL PURPOSE Pellet,

HILLSIDE/CDL Prot- Buffer Meal,

LEE, M.-CLOSE UP PX Pellet,

HIGH DESERT/ GHC LACT Meal,

TATARKA,

M CUST PROT Meal,

SUNRIDGE/CDL PROTEIN Blend,

LOURENZO, K PVM DAIRY Meal,

DOUBLE B DAIRY/GHC LAC Mineral,

WEST PIONT/GHC CLOSEUP Mineral,

WEST POINT/GHC LACT Meal,

JENKS,

J/COMPASS PROTEIN Meal,

COPPINI - 8# SPECIAL DAIRY Mix,

GULICK, L-LACT Meal (Bulk),

TRIPLE J - PROTEIN/LACTATION,

ROCK CREEK/GHC MILK Mineral,

BETTENCOURT/GHC S.SIDE MK-MN,

BETTENCOURT #1/GHC MILK MINR,

V&C DAIRY/GHC LACT Meal,

VEENSTRA, F/GHC LACT Meal,

SMUTNY, A- BYPASS ML W/SMARTA,

Recall # V-025-2007

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

RECALLING FIRM/MANUFACTURER Rangen, Inc, Buhl, ID, by letters on February 13 and 14, 2007.

Firm initiated recall is complete.

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

VOLUME OF PRODUCT IN COMMERCE 9,997,976 lbs.

DISTRIBUTION ID and NV

END OF ENFORCEMENT REPORT FOR MARCH 21, 2007

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

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

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

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

Saturday, August 29, 2009

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

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

Friday, September 4, 2009

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

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

Thursday, March 19, 2009

MILLIONS AND MILLIONS OF POUNDS OF MAD COW FEED IN COMMERCE USA WITH ONGOING 12 YEARS OF DENIAL NOW, WHY IN THE WORLD DO WE TO TALK ABOUT THIS ANYMORE $$$

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

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

*** PLEASE SEE THIS URGENT UPDATE ON CWD AND FEED ANIMAL PROTEIN ***

Sunday, March 20, 2016

Docket No. FDA-2003-D-0432 (formerly 03D-0186) Use of Material from Deer and Elk in Animal Feed ***UPDATED MARCH 2016*** Singeltary Submission

http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/GuidanceforIndustry/UCM052506.pdf?source=govdelivery&utm_medium=email&utm_source=govdelivery

SEE MAD COW FEED VIOLATIONS AFER MAD COW FEED VIOLATIONS ;

http://chronic-wasting-disease.blogspot.com/2016/03/docket-no-fda-2003-d-0432-formerly-03d.html

Tuesday, April 19, 2016

Docket No. FDA-2013-N-0764 for Animal Feed Regulatory Program Standards Singeltary Comment Submission

https://www.regulations.gov/#!documentDetail;D=FDA-2003-D-0432-0011

16 years post mad cow feed ban August 1997 2013

Sunday, December 15, 2013

FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEED VIOLATIONS OFFICIAL ACTION INDICATED OIA UPDATE DECEMBER 2013 UPDATE

http://madcowusda.blogspot.com/2013/12/fda-part-589-substances-prohibited-from.html

Tuesday, December 23, 2014

FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEEDVIOLATIONS OFFICIAL ACTION INDICATED OAI UPDATE DECEMBER 2014 BSE TSE PRION

http://madcowusda.blogspot.com/2014/12/fda-part-589-substances-prohibited-from.html

17 years post mad cow feed ban August 1997

Monday, October 26, 2015

FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEED VIOLATIONS OFFICIAL ACTION INDICATED OIA UPDATE October 2015

http://madcowusda.blogspot.com/2015/10/fda-part-589-substances-prohibited-from.html

TUESDAY, JANUARY 17, 2017

FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEEDVIOLATIONS OFFICIAL ACTION INDICATED OAI UPDATE 2016 to 2017 BSE TSE PRION

http://bovineprp.blogspot.com/2017/01/fda-part-589-substances-prohibited-from.html

TUESDAY, JULY 18, 2017

USDA announces Alabama case of Bovine Spongiform Encephalopathy Alabama

http://bovineprp.blogspot.com/2017/07/usda-announces-alabama-case-of-bovine.html

2012 ATYPICAL L-TYPE BASE BSE TSE PRION CALIFORNIA ‘confirmed’ Saturday, August 4, 2012

*** Final Feed Investigation Summary - California BSE Case - July 2012

http://transmissiblespongiformencephalopathy.blogspot.com/2012/08/final-feed-investigation-summary.html

2009 UPDATE ON ALABAMA AND TEXAS MAD COWS 2005 and 2006

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

THURSDAY, JULY 20, 2017

USDA OIE Alabama Atypical L-type BASE Bovine Spongiform Encephalopathy BSE animal feeds for ruminants rule, 21 CFR 589.200

http://bovineprp.blogspot.com/2017/07/usda-oie-alabama-atypical-l-type-base.html

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.

http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1000156

http://www.plospathogens.org/article/fetchObjectAttachment.action?uri=info%3Adoi%2F10.1371%2Fjournal.ppat.1000156&representation=PDF

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)

http://prionpathy.blogspot.com/2010/08/bse-case-associated-with-prion-protein.html

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

http://www.nature.com/nature/journal/v457/n7233/full/4571079b.html

Thursday, July 24, 2014

*** Protocol for further laboratory investigations into the distribution of infectivity of Atypical BSE SCIENTIFIC REPORT OF EFSA New protocol for Atypical BSE investigations

http://bse-atypical.blogspot.com/2014/07/protocol-for-further-laboratory.html

Thursday, July 24, 2014

*** Protocol for further laboratory investigations into the distribution of infectivity of Atypical BSE SCIENTIFIC REPORT OF EFSA New protocol for Atypical BSE investigations

http://bse-atypical.blogspot.com/2014/07/protocol-for-further-laboratory.html

THURSDAY, FEBRUARY 14, 2013

Unique Properties of the Classical Bovine Spongiform Encephalopathy Strain and Its Emergence From H-Type Bovine Spongiform Encephalopathy Substantiated by VM Transmission Studies

http://bse-atypical.blogspot.com/2013/02/unique-properties-of-classical-bovine.html

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)

http://prionpathy.blogspot.com/2010/08/bse-case-associated-with-prion-protein.html

2009 UPDATE ON ALABAMA AND TEXAS MAD COWS 2005 and 2006

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

http://prionopathy.blogspot.com/2012/03/variably-protease-sensitve-prionopathy.html

P02.35

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

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

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

http://www.neuroprion.org/resources/pdf_docs/conferences/prion2007/abstract_book.pdf

PPo4-15:

A Surprisingly High Number of the Plaque-Like VV sCJD Subtype Among the Polish sCJD-is There a Connection with BASE?

Beata Sikorska and Pawel P. Liberski Department of Molecular Pathology and Neuropathology; Medical University of Lodz; Lodz, Poland

Recently described bovine amyloidotic spongiform encephalopathy (BASE) or L type BSE-was is overrepresented in Poland (15% of all cases of BSE). Moreover, the number of BASE cases in Poland per million bovines is the highest in Europe. A potential human risk from BASE is evident from experimental transmission to "humanized" transgenic animals and primates. Taking into consideration that non-human primate inoculated with BASE had a shorter incubation period than monkeys infected with classical BSE, and that humanized Tg mice have been found to be highly susceptible to infection with atypical form of BSE, it seems probable that BASE may be more pathogenic for humans than BSE, but the transmitted disease may differ from BSE-derived vCJD. Among 47 cases which have been diagnosed as definite in our laboratory, in 19 cases complete histopathological examination and codon 129 status were available. On the basis of the histological pattern and codon 129 status the cases of sCJD were divided into subtypes according to the Parchi&Gambetti classification. The results are as follows: type 1 (MMorMV)- 42%, type 2 (VV)-32%, type 3 (MV)-10.5%, type 4c (MM)- 10.5% and type 5 (VV)-5 %. Although the number of cases is too low to conclude a significantly different distribution of sCJD subtypes in Polish population those data show surprisingly high number of the plaque-like VV sCJD subtype. Interestingly, it was shown before that Tg mice inoculated with BASE showed granular and plaque-like aggregates or PrPSc in brains resembling those observed in VV2 subtype of sCJD.

PPo2-26:

Transmission of Classical and Atypical (L-type) Bovine Spongiform Encephalopathy (BSE) Prions to Cynomolgus macaques

Fumiko Ono,1 Yoshio Yamakawa,2 Minoru Tobiume,3 Yuko Sato,3 Harutaka Katano,3 Kenichi Hagiwara,2 Iori Itagaki,1 Akio Hiyaoka,1 Katuhiko Komatuzaki,1 Yasunori Emoto,1 Hiroaki Shibata,4 Yuichi Murayama,5 Keiji Terao,4 Yasuhiro Yasutomi4 and Tetsutaro Sata3

1The Corporation for Production and Research of Laboratory Primates; Tsukuba City, Japan; 2Departments of Cell Biology and Biochemistry; and 3Pathology; National Institute of Infectious Diseases; Tokyo, Japan; 4Tsukuba Primate Research Center; National Institute of Biomedical Innovation; Tsukuba City, Japan; 5Prion Disease Research Team; National Institute of Animal Health; Tsukuba City, Japan

Key words: L-type BSE, cBSE, cynomolgus macaques, transmission

BSE prion derived from classical BSE (cBSE) or L-type BSE was characterized by inoculation into the brain of cynomolgus macaques. The neurologic manifestation was developed in all cynomolgus macaques at 27-43 months after intracerebral inoculation of brain homogenate from cBSE-affected cattle (BSE JP/6). Second transmission of cBSE from macaque to macaque shortened incubation period to 13-18 months. cBSE-affected macaques showed the similar clinical signs including hyperekplexia, tremor and paralysis in both primary and second transmission.

Two macaques were intracerebrally inoculated brain homogenate from the L-type BSE-affected cattle (BSE JP/24). The incubation periods were 19-20 months in primary transmission.

The clinical course of the L-type BSE-affected macaques differed from that in cBSE-affected macaques in the points of severe myoclonus without hyperekplexia. The glycoform profile of PrPSc detected in macaque CNS was consistent with original pattern of either cBSE or L-typeBSE PrPSc, respectively. Although severe spongiform change in the brain was remarkable in all BSE-affected macaques, severe spongiform spread widely in cerebral cortex in L-type BSE-affected macaques. Heavy accumulation of PrPSc surrounded by vacuola formed florid plaques in cerebral cortex of cBSE-affected macaques. Deposit of PrPSc in L-type BSE-affected macaque was weak and diffuse synaptic pattern in cerebrum, but large PrPSc plaques were evident at cerebellum. MRI analysis, T2, T1, DW and flair sequences, at the time of autopsy revealed that brain atrophy and dilatation of cerebral ventricles were significantly severe in L-type BSE-affected macaques. These results suggest that L-type BSE is more virulent strain to primates comparing to cBSE.

SP1-4:

Evidence from Molecular Strain Typing

Gianluigi Zanusso Department of Neurological and Visual Sciences; Section of Clinical Neurology; University of Verona; Verona, Italy

Key words: molecular analysis, strain typing, atypical BSE, CJD

In 2001, active surveillance for bovine spongiform encephalopathy (BSE) led to the discovery of atypical BSE phenotypes in aged cattle distinct from classical BSE (C-type). These atypical BSE cases had been classified as low L-type (BASE) or high H-type BSE based on the molecular mass and the degree of glycosylation of of the pathological prion protein (PrPSc). Transmission studies in TgBov mice showed that H-type BSE, C-type BSE and BASE behave as distinct prion strains with different incubation periods, PrPSc molecular patterns and pathological phenotypes. A still unclear issue concerns the potential transmissibility and phenotypes of atypical BSEs in humans. We previously indicated that BASE was similar to a distinct subgroup of sporadic form of Creutzfeldt-Jakob disease (sCJD) MV2, based on molecular similarities and on neuropathological pattern of PrP deposition. To investigate a possible link between BASE and sCJD, Kong et al. and Comoy et al. experimentally inoculated TgHu mice (129MM) and a non-human primate respectively, showing in both models that BASE was more virulent compare to BSE. Further, non-human primate reproduced a clinical phenotype resembling to that of sCJD subtype MM2. Here, we presented a comparative analysis of the biochemical fingerprints of PrPSc between the different sCJD subtypes and animal TSEs and after experimental transmission to animals.

http://www.neuroprion.org/resources/pdf_docs/conferences/prion2010/prion_2010_programme.pdf

http://chronic-wasting-disease.blogspot.com/2010/09/cwd-prion-2010.html

http://prionconference.blogspot.com/

8. Even though human TSE‐exposure risk through consumption of game from European cervids can be assumed to be minor, if at all existing, no final conclusion can be drawn due to the overall lack of scientific data.

***> In particular the US data do not clearly exclude the possibility of human (sporadic or familial) TSE development due to consumption of venison.

The Working Group thus recognizes a potential risk to consumers if a TSE would be present in European cervids. It might be prudent considering appropriate measures to reduce such a risk, e.g. excluding tissues such as CNS and lymphoid tissues from the human food chain, which would greatly reduce any potential risk for consumers.. However, it is stressed that currently, no data regarding a risk of TSE infections from cervid products are available.

snip...

The tissue distribution of infectivity in CWD‐infected cervids is now known to extend beyond CNS and lymphoid tissues. While the removal of these specific tissues from the food chain would reduce human dietary exposure to infectivity, exclusion from the food chain of the whole carcass of any infected animal would be required to eliminate human dietary exposure.

https://efsa.onlinelibrary.wiley.com/doi/full/10.2903/j.efsa.2018.5132

PLOS ONE Journal

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

https://journals.plos.org/plosone/article/comment?id=10.1371/annotation/4f9be886-69fe-4c7c-922b-85b0ecbe6d53

IBNC Tauopathy or TSE Prion disease, it appears, no one is sure

Terry S. Singeltary Sr., 03 Jul 2015 at 16:53 GMT

***however in 1 C-type challenged animal, Prion 2015 Poster Abstracts S67 PrPsc was not detected using rapid tests for BSE.

***Subsequent testing resulted in the detection of pathologic lesion in unusual brain location and PrPsc detection by PMCA only.

*** IBNC Tauopathy or TSE Prion disease, it appears, no one is sure ***

Greetings Plos et al,

in reference to;

‘’A Naturally Occurring Bovine Tauopathy Is Geographically Widespread in the UK’’

I kindly wish to comment please, as follows.

I was stunned by this report.

This Research Report should have been titled ;

‘’It Appears A Naturally Occurring Bovine Tauopathy Is Geographically Widespread in the UK’’

Thus IBNC _appears_ to be the first naturally occurring and geographically widespread tauopathy in a species other than humans.<<<

IBNC _appear_ to lack intracellular neurofibrillary tangles,<<<

Thus IBNC _appears_ to be the first naturally occurring and geographically widespread tauopathy in a species other than humans<<<

IBNC appears to be a complex proteinopathy with evidence for additional secondary accumulation of alpha synuclein, ubiquitin, and possibly PrP [5]. <<<

however, it appears to represent a naturally-occurring predominantly 3R tauopathy in a non-primate species.<<<

In the absence of any specific aetiological cause of IBNC yet identified, and based on the existing epidemiology and presence of P-tau we have considered the possibility that IBNC might also have a significant environmental component. Epidemiological investigations to explore usage of herbicides, insecticides, parasiticides and other chemicals used in agriculture would be helpful in future investigations of IBNC and might provide further insight into some causes of neurodegeneration in man.<<<

This is supposition at best in my opinion, and at worse, I will refrain from comment.

Too many _appears_ , appears this, appears that.

Too many _In the absence of any specific aetiological cause of IBNC yet identified_

is this what science has come to?

has science become some entwined with corporate special interest, do we change science now?

I believe that is far too early to rule out IBNC as a Transmissible Spongiform Encephalopathy TSE prion disease, either of typical or typical strain.

it can be argued that there is potential for Alzheimer’s to be a low dose TSE Prion disease ;

https://journals.plos.org/plosone/article/comment?id=10.1371/annotation/5adef4ac-a7e4-46a4-8806-c8533d5c862c

***> 2026 Transmissible Spongiform Encephalopathy TSE, CWD, BSE, Scrapie, Pig Prion PrP <***

Chronic wasting disease prions in cervids and wild pigs in North America Preliminary Outbreak Assessment DEFRA 26 January 2026 Department for Environment, Food and Rural Affairs

Preliminary Outbreak Assessment

Chronic wasting disease prions in cervids and wild pigs in North America

26 January 2026

Disease report

Chronic wasting disease (CWD) is a fatal neurodegenerative disease of cervids, such as deer, elk, moose and reindeer. It is caused by prions – infectious proteins that cause normal cellular prion proteins to misfold (CIDRAP, 2025). The disease is widespread in captive and free-ranging cervids in North America (Figure 1). For the first time, CWD prions have also been detected in the tissues of wild pigs (Sus scrofa) caught in CWD-affected areas of the USA (Soto et al. 2025). This discovery emerged from a study designed to investigate potential interactions between wild pigs and CWD prions, as wild pigs often coexist with cervids, which can shed prions into the environment. The following assessment discusses the epidemiology of CWD in North America and the detection of CWD prions in wild pigs. It also considers the potential implications for Great Britain.

Conclusion

CWD has continued to spread among captive and free-ranging cervids in North America since it was first detected in the 1960s. The finding of CWD prions in wild pigs in the USA suggests they could contribute towards transmission of the disease, influencing its epidemiology, geographic distribution and interspecies spread. However, further research is needed to confirm this. CWD has never been reported in Great Britain and the current risk of CWD prions being introduced into Great Britain’s wild pig or cervid population ranges from very low to negligible.

Readers are reminded to be vigilant for signs of CWD. Information on how to spot the disease can be found here. Suspected cases must be reported immediately to the Defra Rural Services Helpline on 03000 200 301. In Wales, call 0300 303 8268. In Scotland, contact your local Field Services Office. Failure to do so is an offence. We will continue to monitor the situation.

Authors • Lawrence Finn • Dr. Lauren Perrin • John Spiropoulos • Dr. Helen RobertsDepartment for Environment, Food and Rural Affairs

References

snip…see;

https://assets.publishing.service.gov.uk/media/697a3b013c71d838df6bd413/CWD_Prions_in_Cervids_and_Wild_Pigs_in_North_America.pdf

Detection of Prions in Wild Pigs (Sus scrofa) from Areas with Reported Chronic Wasting Disease Cases, United State Volume 31, Number 1—January 2025

https://wwwnc.cdc.gov/eid/article/31/1/24-0401_article

THURSDAY, JANUARY 08, 2026

Confucius Ponders, what about Wild Pigs (Sus scrofa) and CWD TSE Prion, and the Environment, what if?

Confucius Ponders, what about Wild Pigs (Sus scrofa), they can cover some distance rather quickly, what about Wild Pigs (Sus scrofa) digging up the terrain, and as they do it, what if these Wild Pigs (Sus scrofa) were exposed to CWD TSE Prion, and then they go on exposing and saturating the land with CWD TSE Prion, then the soil becomes contaminated with CWD TSE Prion, then what about the plants that grow from that soil for the decades to come, what if???

https://prpsc.proboards.com/thread/190/confucius-ponders-wild-pigs-scrofa

https://chronic-wasting-disease.blogspot.com/2026/01/confucius-ponders-what-about-wild-pigs.html

'Spontaneous mutation'

***Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility.***

Even if the prevailing view is that sporadic CJD is due to the spontaneous formation of CJD prions, it remains possible that its apparent sporadic nature may, at least in part, result from our limited capacity to identify an environmental origin.

https://www.nature.com/articles/srep11573

***> US Report, Scrapie, CWD, Cattle, Sheep, Pigs, Cervid, Humans, Zoonotic, 2026 <***

*** Grant Agreement number: 222887 ***

*** Project acronym: PRIORITY ***

*** Project title: Protecting the food chain from prions: shaping European priorities through basic and applied research Funding ***

Scheme: Large-scale integrating project Period covered: from Oct. 1, 2009 to Sept. 30, 2014

Name of the scientific representative of the project's co-ordinator1, Title and Organisation: Jesús R. Requena, Ph.D., Associate Professor, Department of medicine, University of Santiago de Compostela, Spàin. Tel: 34-881815464 Fax: 34-881815403 E-mail: jesus.requena@usc.es

Project website¡ Error! Marcador no definido. address: www.prionpriority.eu

PRIORITY, PROJECT FINAL REPORT

*** 14) Concluding that atypical scrapie can transmit to Humans and that its strain properties change as it transmits between species ***

snip...

http://cordis.europa.eu/docs/results/222/222887/final1-priority-final-report.pdf

see;

https://nor-98.blogspot.com/2016/09/goat-k222-prpc-polymorphic-variant-does.html

Block D: Prion epidemiology

Studies on atypical scrapie were identified as a key element of this block, given the potential risk associated to this agent. We studied the permeability of Human, bovine and porcine species barriers to atypical scrapie agent transmission. Experiments in transgenic mice expressing bovine, porcine or human PrPC suggest that this TSE agent has the intrinsic ability to propagate across these species barriers including the Human one. Upon species barrier passage the biological properties and phenotype of atypical scrapie seem to be altered. Further experiments are currently ongoing (in the framework of this project but also in other projects) in order to: (i) characterize the properties of the prion that emerged from the propagation of atypical scrapie in tg Hu; (ii) to confirm that the phenomena we observed are also true for atypical scrapie isolates other than the ones we have studied.

In parallel, studies in sheep have concluded that:

*** Atypical scrapie can be transmitted by both oral and intracerebral route in sheep with various PRP genotypes

*** Low but consistent amount of infectivity accumulates in peripheral tissue (mammary gland, lymph nodes, placenta, skeletal muscles, nerves) of sheep incubating atypical scrapie.

*** The combination of data from all our studies leads us to conclude that:

*** Atypical scrapie passage through species barriers can lead to the emergence of various prions including classical BSE (following propagation in porcine PRP transgenic mice).

*** Atypical scrapie can propagate, with a low efficacy, in human PrP expressing mice. This suggests the existence of a zoonotic potential for this TSE agent.

snip...

We advance our main conclusions and recommendations, in particular as they might affect public policy, including a detailed elaboration of the evidence that led to them. Our main recommendations are:

a. The issue of re-introducing ruminant protein into the food-chain The opinion of the members of Priority is that sustaining an absolute feed ban for ruminant protein to ruminants is the essential requirement, especially since the impact of non-classical forms of scrapie in sheep and goats is not fully understood and cannot be fully estimated. Therefore, the consortium strongly recommends prohibiting re-introduction of processed ruminant protein into the food-chain. Arguments in support of this opinion are:

• the large (and still uncharacterized) diversity of prion agents that circulate in animal populations;

• the uncertainties related to prion epidemiology in animal populations;

• the unknown efficacy of industrial processes applied to reduce microbiological risk during processed animal protein (PAP) production on most prion agents; • the intrinsic capacity of prions to cross interspecies transmission barriers; • the lack of sensitive methodology for identifying cross contamination in food.

• the evolution of natural food chains in nature (i.e. who eats whom or what) has generated an efficient barrier preventing, to some extent, novel prion epidemies and that this naturally evolved ecology should be respected.

The consortium is also hesitant to introduce processed ruminant proteins into fish food considering the paucity of data on prion infections in fishes and sea animals including those of mammalian origin, and the risk of establishing an environmental contamination of the oceans that cannot be controlled.

b. Atypical prion agents and surveillance

Atypical prion agents (see below) will probably continue to represent the dominant form of prion diseases in the near future, particularly in Europe.

*** Atypical L-type BSE has clear zoonotic potential, as demonstrated in experimental models.

*** Similarly, there are now some data that seem to indicate that the atypical scrapie agent can cross various species barriers.

*** Moreover, the current EU policy for eradicating scrapie (genetic selection in affected flocks) is ineffective for preventing atypical scrapie.

*** The recent identification of cell-to-cell propagation and the protein-encoded strain properties of human neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, suggest that they bear the potential to be transmissible even if not with the same efficiency as CJD. More epidemiological data from large cohorts are necessary to reach any conclusion on the impact of their transmissibility on public health. Re-evaluations of safety precautions may become necessary depending on the outcome of these studies. In that context it would appear valuable

• to develop knowledge related to the pathogenesis and inter-individual transmission of atypical prion agents in ruminants (both intra- and inter-species)

• to improve the sensitivity of detection assays that are applied in the field towards this type of agent

• to maintain a robust surveillance of both animal and human populations

c. The need for extended research on prions

Intensified searching for a molecular determinants of the species barrier is recommended, since this barrier is a key for many important policy areas - risk assessment, proportional policies, the need for screening of human products and food. In this respect, prion strain structural language also remains an important issue for public health for the foreseeable future. Understanding the structural basis for strains and the basis for adaptation of a strain to a new host will require continued fundamental research. Prions maintain a complex two-way relationship with the host cell and fundamental research is needed on mechanisms for their transmission, replication and cause of nervous system dysfunction and death.

Early detection of prion infection, ideally at preclinical stage, also remains crucial for development of effective treatment strategies in humans affected by the disease.

Position of the Priority consortium

Nearly 30 years ago, the appearance in the UK of Bovine Spongiform Encephalopathy (BSE) quickly brought the previously obscure “prion diseases” to the spotlight. The ensuing health and food crises that spread throughout Europe had devastating consequences. In the UK alone, there were more than 36,000 farms directly affected by BSE and the transmission of BSE prions to humans via the food chain has caused over 200 people in Europe to die from variant Creutzfeldt-Jakob disease (vCJD) (http://www.cjd.ed.ac.uk

Origins of prion epidemies

Classical BSE now appears to be under control, with 18 EU Member States having achieved the World Organisation for Animal Health (Office International Epizooties) „negligible risk‟ status (May 2014; http://www.oie.int/en/animal-health-in-the-world/official-disease-status/bse/list-of-bse-risk-status/), and the remaining MS assessed as „controlled‟ risk. Of note, research, including EU-funded research, has played a key role in this success: while the origin of the infection was never defined, the principle driver of the epidemic was identified as prions in Meat and Bone Meal (MBM). Tests based on prion protein-specific antibodies were developed, allowing detection of infected animals, and a better understanding of disease pathogenesis and the distribution of infectivity in edible tissues; experimental investigation of transmission barriers between different species allowed a rational estimation of risks, etc. All of this led to the implementation of rational and effective policies, such as the MBM ban to protect the animal feed chain, and the Specified Risk Material (SRM) regulations to protect the human food chain.

In spite of this progress, prions are still a threat. Epidemiological re-assessment indicates that the ∼10 year incubation period separating the peaks of the BSE and the vCJD epidemics is probably too short. In addition, results from a large number of human tonsil and appendix analyses in the UK suggest that there may be a high number of asymptomatic individuals who are positive for the disease-associated conformer prion protein PrPSc. While vCJD is the only form of human prion disease that has been consistently demonstrated to have lymphoreticular involvement, there has been no systematic investigation of lymphoid tissue in cases with other prion diseases.

The human prion problem

The clinical cases of vCJD identified to date have all shared a common PrP genotype (M129M), although one pre-clinical case was confirmed as an M129V heterozygote, and it has been mooted that perhaps only the M129M proportion of the population is susceptible. However, in the UK appendix study, PrP accumulation was described in samples representing every codon 129 genotype, raising the possibility that genotype does not confer resistance but instead modulates incubation period. Apart from the two UK studies, the lymphoid tissues of non-CJD patients have not been examined for the presence of PrPSc, so, these cases may not solely represent pre-clinical vCJD, but also other forms of prion disease.

Recent experiments in highly susceptible mouse models indicate the presence of infectivity in blood or blood components at late disease stages in sporadic CJD. The significance of this experimental finding for humans has to be explored in more detail and, at the present time, there is no evidence for the transmission of prions via blood in sporadic CJD. However a likely scenario is that all those with signs of infection or abnormal PrP accumulation in peripheral tissue could have infective blood, posing the risk for transmission via blood products, which has been clearly demonstrated in experimental models, and confirmed in several cases of vCJD in man. Altogether, these data clearly demonstrate the potential risk of a second wave of vCJD, particularly when the number people identified with lymphoid accumulation of PrPSc (16/32,411) gives a prevalence estimate in the UK of 493 per million, much higher than the number of clinical cases seen to date.

The animal prion problem

An increasing number of reports on cases of “atypical” BSE in cattle throughout the EU and beyond may lead to a new epidemic, particularly since we still do not understand all factors determining the species barrier. Ovine scrapie is another concern, because it could mask ovine BSE, presumably transmissible to humans. Scrapie is endemic and not likely to be eradicated soon, although current control measures are effective at greatly reducing disease incidence. Atypical forms, which may be spontaneous, are not affected by these control measures and these forms of disease will persist in the global animal population. The low prevalence of these disease forms makes effective surveillance very challenging. However, there is a clear risk attendant on ignoring these cases without an understanding of their possible zoonotic potential, particularly when most forms of human disease have no established aetiology. In summary, atypical cases of BSE and scrapie presently clearly outnumber classical cases in cattle and sheep in all member states.

We will highlight the state-of-the-art knowledge and point out scientific challenges and the major questions for research. Strategic objectives and priorities in Europe in the future for research that aims to control, eliminate or eradicate the threat posed by prions to our food and health are also indicated.

The Priority project has focused on 4 themes, namely the structure, function, conversion and toxicity of prions; detection of prions; mechanisms of prion transmission and spreading and epidemiology of prion diseases. This paper summarizes the opinions/positions reached within these themes at the end of the project.

http://cordis.europa.eu/docs/results/222/222887/final1-priority-final-report.pdf

see;

https://nor-98.blogspot.com/2016/09/goat-k222-prpc-polymorphic-variant-does.html

Transmission of scrapie prions to primate after an extended silent incubation period

*** In complement to the recent demonstration that humanized mice are susceptible to scrapie, we report here the first observation of direct transmission of a natural classical scrapie isolate to a macaque after a 10-year incubation period. Neuropathologic examination revealed all of the features of a prion disease: spongiform change, neuronal loss, and accumulation of PrPres throughout the CNS.

*** This observation strengthens the questioning of the harmlessness of scrapie to humans, at a time when protective measures for human and animal health are being dismantled and reduced as c-BSE is considered controlled and being eradicated.

*** Our results underscore the importance of precautionary and protective measures and the necessity for long-term experimental transmission studies to assess the zoonotic potential of other animal prion strains.

http://www.ars.usda.gov/research/publications/publications.htm?SEQ_NO_115=313160

O.05: Transmission of prions to primates after extended silent incubation periods: Implications for BSE and scrapie risk assessment in human populations

*** We recently observed the direct transmission of a natural classical scrapie isolate to macaque after a 10-year silent incubation period,

***with features similar to some reported for human cases of sporadic CJD, albeit requiring fourfold long incubation than BSE. Scrapie, as recently evoked in humanized mice (Cassard, 2014),

***is the third potentially zoonotic PD (with BSE and L-type BSE),

***thus questioning the origin of human sporadic cases.

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

PRION 2015 CONFERENCE

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5019500/

PRION 2016 TOKYO

Saturday, April 23, 2016

SCRAPIE WS-01: Prion diseases in animals and zoonotic potential 2016

Prion. 10:S15-S21. 2016 ISSN: 1933-68961933-690X

WS-01: Prion diseases in animals and zoonotic potential

***Transmission data also revealed that several scrapie prions propagate in HuPrP-Tg mice with efficiency comparable to that of cattle BSE. While the efficiency of transmission at primary passage was low, subsequent passages resulted in a highly virulent prion disease in both Met129 and Val129 mice.

***Transmission of the different scrapie isolates in these mice leads to the emergence of prion strain phenotypes that showed similar characteristics to those displayed by MM1 or VV2 sCJD prion.

***These results demonstrate that scrapie prions have a zoonotic potential and raise new questions about the possible link between animal and human prions.

http://www.tandfonline.com/doi/abs/10.1080/19336896.2016.1163048?journalCode=kprn20

Title: Transmission of scrapie prions to primate after an extended silent incubation period)

*** In complement to the recent demonstration that humanized mice are susceptible to scrapie, we report here the first observation of direct transmission of a natural classical scrapie isolate to a macaque after a 10-year incubation period. Neuropathologic examination revealed all of the features of a prion disease: spongiform change, neuronal loss, and accumulation of PrPres throughout the CNS.

*** This observation strengthens the questioning of the harmlessness of scrapie to humans, at a time when protective measures for human and animal health are being dismantled and reduced as c-BSE is considered controlled and being eradicated.

*** Our results underscore the importance of precautionary and protective measures and the necessity for long-term experimental transmission studies to assess the zoonotic potential of other animal prion strains.

http://www.ars.usda.gov/research/publications/publications.htm?SEQ_NO_115=313160

Comparing the Distribution of Ovine Classical Scrapie and Sporadic Creutzfeldt-Jakob Disease in Italy: Spatial and Temporal Associations (2002-2014)

Ru G1 ., Pocchiari M2 ., Bertolini S. 1, Pite L.1 , Puopolo M.2 , Ladogana A.2 , Perrotta M.G.3 , Meloni D 1 . (1) National reference center for the study and research on animal encephalopathies and comparative neuropathologies (CEA). Experimental Zooprophylactic Institute of Piemonte, Liguria and Valle d'Aosta, Torino, Italy.

(2) Department of Cellular Biology and Neuroscience, Istituto Superiore di Sanità, Roma, Italy. (3) Office 3 National center for the fight and emergency against animal diseases. Ministry of Health, Roma, Italy.

Aim: This study aims to investigate potential spatial and temporal associations between Creutzfeldt-Jakob disease (CJD) in humans (2010-2014) and ovine classical scrapie (CS) (2002- 2006) in Italy, serving as a proxy for exposure.

Materials and Methods: National data from prion disease surveillance in humans (sporadic CJD) and small ruminants (CS) in Italy were utilized. A descriptive geographic analysis was conducted for each disease individually. Subsequently, an ecological study was performed to compare the occurrence of both diseases at the district and regional levels. Standardized incidence ratios (SIR), adjusted for confounders, were calculated for CJD and CS by district and region, respectively, representing the outcome and proxy of exposure. Considering a possible long incubation period of CJD, two study periods were analysed: 2010-2014 for CJD and 2002-2006 for CS. Eight alternative linear regression models were developed using SIR in humans as the dependent variable and SIR in sheep as the independent variable. These models varied in the scale of SIR data (continuous vs. categorical), geographical level (district vs. region), and the potential past exposure of sheep in specific areas to a known source of infection (via a contaminated vaccine).

Results: The analysis of data at the district level revealed no significant association. However, when considering aggregated regional data, all four models consistently indicated a statistically significant positive association, suggesting a higher incidence of the disease in humans as the regional incidence of sheep scrapie increased.

Conclusions: While the results are intriguing, it is important to acknowledge the inherent limitations of ecological studies. Nevertheless, these findings provide valuable evidence to formulate a hypothesis regarding the zoonotic potential of classical scrapie. Further investigations are necessary, employing specific designs such as analytical epidemiology studies, to test this hypothesis effectively.

Funded by: Italian Ministry of Health Grant number: Realizzazione del programma epidemiologico finalizzato a dare evidenza del potenziale zoonotico delle TSE animali diverse dalla BSE. Prot. N. 0018730-17/07/2015-DGSAFCOD_UO-P

''Nevertheless, these findings provide valuable evidence to formulate a hypothesis regarding the zoonotic potential of classical scrapie. Further investigations are necessary, employing specific designs such as analytical epidemiology studies, to test this hypothesis effectively.''

Meeting-book-final-version prion 2023 Prion 2023 Congress Organizing Committee and the NeuroPrion Association, we invite you to join us for the International Conference Prion2023 from 16-20 October 2023 in Faro, Portugal.

https://prion2023.org/wp-content/uploads/2023/10/Meeting-book-final-version2.pdf

https://web.archive.org/web/20250828201533/https://prion2023.org/wp-content/uploads/2023/10/Meeting-book-final-version2.pdf

https://www.researchgate.net/profile/Syed-Zahid-Shah/publication/378314391_Meeting-book-final-version_prion_2023/links/65d44dad28b7720cecdca95f/Meeting-book-final-version-prion-2023.pdf

***> CWD Action Plan National Program 103 Animal Health 2022-2027 UPDATE JANUARY 2026

https://prpsc.proboards.com/thread/189/action-national-program-animal-health

https://chronic-wasting-disease.blogspot.com/2026/01/cwd-action-plan-national-program-103.html

***> SCRAPIE TSE Prion USA RAPID RESPONSE URGENT UPDATES DECEMBER 25, 2025

***> CWD vs Scrapie Urgent Update

https://scrapie-usa.blogspot.com/2025/12/scrapie-tse-prion-usa-rapid-response.html

https://prpsc.proboards.com/thread/186/scrapie-prion-response-urgent-updates

***> 2026 USDA EXPLANATORY NOTES, APHIS, CWD, BSE, Scrapie, TSE, Prion

https://transmissiblespongiformencephalopathy.blogspot.com/2025/12/2026-usda-explanatory-notes-aphis-cwd.html

Research Project: Elucidating the Pathobiology and Transmission of Transmissible Spongiform Encephalopathies

Location: Virus and Prion Research

2025 Annual Report

https://transmissiblespongiformencephalopathy.blogspot.com/2025/12/ars-research-elucidating-pathobiology.html

USDA National Scrapie Program History and Bovine Spongiform Encephalopathy BSE TSE Update 2025 and history there from

https://www.researchgate.net/publication/396084947_USDA_National_Scrapie_Program_History_and_Bovine_Spongiform_Encephalopathy_BSE_TSE0AUpdate_2025

APHIS USDA Captive CWD Herds Update by State December 2025 Update

https://chronic-wasting-disease.blogspot.com/2025/12/aphis-usda-captive-cwd-herds-update-by.html

https://prpsc.proboards.com/thread/187/aphis-captive-herds-update-december

TUESDAY, SEPTEMBER 30, 2025

USDA EXPLANATORY NOTES ANIMAL AND PLANT HEALTH INSPECTION SERVICE 2025-2014 CHRONIC WASTING DISEASE CWD TSE CERVID

https://chronic-wasting-disease.blogspot.com/2025/09/usda-explanatory-notes-animal-and-plant.html

TUESDAY, SEPTEMBER 30, 2025

USDA National Scrapie Program History and Bovine Spongiform Encephalopathy BSE TSE Update 2025

https://bovineprp.blogspot.com/2025/09/usda-national-scrapie-program-history.html

TUESDAY, SEPTEMBER 30, 2025

USDA National Scrapie Program History and Bovine Spongiform Encephalopathy BSE TSE Update 2025

https://scrapie-usa.blogspot.com/2025/09/usda-national-scrapie-program-history.html

US NATIONAL PRION DISEASE PATHOLOGY SURVEILLANCE CENTER CJD TSE REPORT SEPTEMBER 2025

From the first full year of reporting CJD TSE in the US in 2000, where 90 cases of CJD was reported that year, to today, where in September 2025, the number of CJD cases reported in the last full year reporting, which would have been 2024, the number of CJD cases for 2024 was 249 cases. So, from the first full year 2000 CJD cases were 90 cases confirmed in that year, to 2024, where 2024 CJD statistics rose to 249 confirmed CJD cases in a single year. A dramatic increase in deaths, from figures that don’t seem to be dramatic. But thes figures today, they are not from “better surveillance”, that dog don’t hunt no more. They have been saying this for over 25 years, year after year, well it’s time to call it for what it is, Human Transmissible Spongiform Encephalopathy TSE Prion cases are rising, and it’s NOT because of better surveillance, or just a “happenstance of bad luck, that 85%+ of all human cases, sporadic CJD, including VPSPr, just happen spontaneously, no, it’s because of unknown environmental factors, and or iatrogenic factors, imho…terry

US NATIONAL PRION DISEASE PATHOLOGY SURVEILLANCE CENTER CJD TSE REPORT SEPTEMBER 2025

https://prionunitusaupdate.blogspot.com/2025/10/us-national-prion-disease-pathology.html

So, this is what we leave our children and grandchildren?

2001 Singeltary on CJD, Journal of American Medical Association

February 14, 2001

Diagnosis and Reporting of Creutzfeldt-Jakob Disease

Terry S. Singeltary, Sr

Author Affiliations

JAMA. 2001;285(6):733-734. doi:10-1001/pubs.JAMA-ISSN-0098-7484-285-6-jlt0214

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

February 14, 2001

Diagnosis and Reporting of Creutzfeldt-Jakob Disease

Terry S. Singeltary, Sr

Author Affiliations

JAMA. 2001;285(6):733-734. doi:10-1001/pubs.JAMA-ISSN-0098-7484-285-6-jlt0214

https://jamanetwork.com/journals/jama/article-abstract/1031186

RE-Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States 2003 Singeltary Journal of Neurology

26 MARCH 2003

RE-Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States

Terry S. Singeltary, retired (medically)

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

https://www.neurology.org/doi/10.1212/01.WNL.0000036913.87823.D6

14th ICID International Scientific Exchange Brochure -

Final Abstract Number: ISE.114

Session: International Scientific Exchange

Transmissible Spongiform encephalopathy (TSE) animal and human TSE in North America

update October 2009

T. Singeltary

Bacliff, TX, USA

Background:

An update on atypical BSE and other TSE in North America. Please remember, the typical U.K. c-BSE, the atypical l-BSE (BASE), and h-BSE have all been documented in North America, along with the typical scrapie's, and atypical Nor-98 Scrapie, and to date, 2 different strains of CWD, and also TME. All these TSE in different species have been rendered and fed to food producing animals for humans and animals in North America (TSE in cats and dogs ?), and that the trading of these TSEs via animals and products via the USA and Canada has been immense over the years, decades.

Methods:

12 years independent research of available data

Results:

I propose that the current diagnostic criteria for human TSEs only enhances and helps the spreading of human TSE from the continued belief of the UKBSEnvCJD only theory in 2009. With all the science to date refuting it, to continue to validate this old myth, will only spread this TSE agent through a multitude of potential routes and sources i.e. consumption, medical i.e., surgical, blood, dental, endoscopy, optical, nutritional supplements, cosmetics etc.

Conclusion:

I would like to submit a review of past CJD surveillance in the USA, and the urgent need to make all human TSE in the USA a reportable disease, in every state, of every age group, and to make this mandatory immediately without further delay. The ramifications of not doing so will only allow this agent to spread further in the medical, dental, surgical arena's. Restricting the reporting of CJD and or any human TSE is NOT scientific. Iatrogenic CJD knows NO age group, TSE knows no boundaries. I propose as with Aguzzi, Asante, Collinge, Caughey, Deslys, Dormont, Gibbs, Gajdusek, Ironside, Manuelidis, Marsh, et al and many more, that the world of TSE Transmissible Spongiform Encephalopathy is far from an exact science, but there is enough proven science to date that this myth should be put to rest once and for all, and that we move forward with a new classification for human and animal TSE that would properly identify the infected species, the source species, and then the route.

http://ww2.isid.org/Downloads/14th_ICID_ISE_Abstracts.pdf

archived url;

https://web.archive.org/web/20100216162542/http://ww2.isid.org/Downloads/14th_ICID_ISE_Abstracts.pdf

International Society for Infectious Diseases Web: http://www.isid.org

http://prionunitusaupdate2008.blogspot.com/2010/01/human-prion-diseases-in-united-states.html

my comments to PLosone here ;

http://www.plosone.org/annotation/listThread.action?inReplyTo=info%3Adoi%2F10.1371%2Fannotation%2F04ce2b24-613d-46e6-9802-4131e2bfa6fd&root=info%3Adoi%2F10.1371%2Fannotation%2F04ce2b24-613d-46e6-9802-4131e2bfa6fd

Wednesday, February 24, 2010

Transmissible Spongiform encephalopathy (TSE) animal and human TSE in North America 14th

ICID International Scientific Exchange Brochure -

http://transmissiblespongiformencephalopathy.blogspot.com/2010/02/transmissible-spongiform-encephalopathy.html

2023

https://creutzfeldt-jakob-disease.blogspot.com/2023/09/professor-john-collinge-on-tackling.html

***> NOW think iatrogenic TSE PrP transmission to humans from exposure to Cervid CWD TSE PrP, what if?

all iatrogenic cjd is, is sporadic cjd, before the iatrogenic event is discovered, traced back, proven, documented, put into the academic domain, and then finally the public domain, this very seldom happens, thus problem solved, it's all sporadic cjd.

iatrogenic Transmissible Spongiform Encephalopathy

https://itseprion.blogspot.com/

UK Jonh Collinge MRC on sporadic CJD;

“sporadic Creutzfeldt-Jakob disease (sCJD), a rapidly progressive dementia which accounts for around 1 in 5000 deaths worldwide.”

***> 2023 Professor John Collinge on tackling prion diseases <***

“The best-known human prion disease is sporadic Creutzfeldt-Jakob disease (sCJD), a rapidly progressive dementia which accounts for around 1 in 5000 deaths worldwide.”

There is accumulating evidence also for iatrogenic AD.

Understanding prion biology, and in particular how propagation of prions leads to neurodegeneration, is therefore of central research importance in medicine.

https://www.ucl.ac.uk/brain-sciences/research/dementia-ucl-priority/professor-john-collinge-tackling-prion-diseases

WEDNESDAY, OCTOBER 15, 2025

US NATIONAL PRION DISEASE PATHOLOGY SURVEILLANCE CENTER CJD TSE REPORT 2025

https://prionunitusaupdate.blogspot.com/2025/10/us-national-prion-disease-pathology.html

FRIDAY, NOVEMBER 21, 2025

While no one was watching: Tenuous status of CDC prion unit, risk of CWD to people worry scientists

https://chronic-wasting-disease.blogspot.com/2025/11/while-no-one-was-watching-tenuous.html

SATURDAY, JANUARY 10, 2026

Neuropsychiatric symptoms in sporadic Creutzfeldt-Jakob disease, a review

https://creutzfeldt-jakob-disease.blogspot.com/2026/01/neuropsychiatric-symptoms-in-sporadic.html

https://prpsc.proboards.com/thread/191/neuropsychiatric-symptoms-sporadic-cjd-review

Cattle with the E211K polymorphism, and gCJD linked to a glutamic acid to lysine substitution at codon 200 (E200K) of PRNP, what if?

https://creutzfeldt-jakob-disease.blogspot.com/2026/01/cattle-with-e211k-polymorphism-and-gcjd.html

Cattle with the E211K vs Humans E200K of PRNP, what if?

https://prpsc.proboards.com/thread/195/cattle-e211k-humans-e200k-prnp

terry