Saturday, August 14, 2010
BSE Case Associated with Prion Protein Gene Mutation (g-h-BSEalabama) and
VPSPr PRIONPATHY
BSE Case Associated with Prion Protein Gene Mutation
Ju¨ rgen A. Richt1¤*, S. Mark Hall2 1 National Animal Disease Center,
United States Department of Agriculture, Agriculture Research Service, Ames,
Iowa, United States of America, 2 National Veterinary Services Laboratories,
Pathobiology Laboratory, Animal and Plant Health Inspection Service, United
States Department of Agriculture, Ames, Iowa, United States of America
Abstract
Bovine spongiform encephalopathy (BSE) is a transmissible spongiform
encephalopathy (TSE) of cattle and was first detected in 1986 in the United
Kingdom. It is the most likely cause of variant Creutzfeldt-Jakob disease (CJD)
in humans. The origin of BSE remains an enigma. Here we report an H-type BSE
case associated with the novel mutation E211K within the prion protein gene
(Prnp). Sequence analysis revealed that the animal with H-type BSE was
heterozygous at Prnp nucleotides 631 through 633. An identical pathogenic
mutation at the homologous codon position (E200K) in the human Prnp has been
described as the most common cause of genetic CJD. This finding represents the
first report of a confirmed case of BSE with a potential pathogenic mutation
within the bovine Prnp gene. A recent epidemiological study revealed that the
K211 allele was not detected in 6062 cattle from commercial beef processing
plants and 42 cattle breeds, indicating an extremely low prevalence of the E211K
variant (less than 1 in 2000) in cattle.
Citation: Richt JA, Hall SM (2008) BSE Case Associated with Prion Protein
Gene Mutation. PLoS Pathog 4(9): e1000156. doi:10.1371/journal.ppat.1000156
Editor: David Westaway, University of Alberta, Canada
Received June 5, 2008; Accepted August 15, 2008; Published September 12,
2008
This is an open-access article distributed under the terms of the Creative
Commons Public Domain declaration which stipulates that, once placed in the
public domain, this work may be freely reproduced, distributed, transmitted,
modified, built upon, or otherwise used by anyone for any lawful purpose.
Funding: This work was supported by the USDA-ARS-National Animal Disease Center
(NADC) and USDA-APHIS-National Veterinary Services Laboratories (NVSL) and by
the NIAID-NIH PO1 AI 77774-01 ‘‘Pathogenesis, Transmission and Detection of
Zoonotic Prion Diseases’’.
Competing Interests: Patent pending: Dr. Ju¨ rgen A. Richt submitted a
patent application entitled ‘‘Novel Polymorphism in Bovine Prion Protein Gene
Sequence’’ (Docket Number 0078.06; Serial No. 11/787,784) on April 18,
2006.
* E-mail: jricht@vet.k-state.edu
¤ Current address: Kansas State University, College of Veterinary Medicine,
DM/P, Manhattan, Kansas, United States of America
Author Summary
Bovine spongiform encephalopathy (BSE or Mad Cow Disease), a transmissible
spongiform encephalopathy (TSE) or prion disease of cattle, was first discovered
in the United Kingdom in 1986. BSE is most likely the cause of a human prion
disease known as variant Creutzfeldt Jakob Disease (vCJD). 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.
Introduction
Transmissible spongiform encephalopathy (TSE) agents induce fatal
neurodegenerative diseases in humans and in some mammalian species [1].
According to the prion-only hypothesis, infectious prions are composed of an
abnormal isoform of a hostencoded glycoprotein, called prion protein (PrPc). The
diseaseassociated form, PrPd, is derived from PrPc by a post-translational
mechanism that involves conformational change [1]. Human TSEs include
Creutzfeldt–Jakob disease (CJD), Gerstmann– Stra¨ussler–Scheinker syndrome, Kuru
and Fatal Familial Insomnia [1]. In animals, several distinct TSE diseases are
recognized: Scrapie in sheep and goats, transmissible mink encephalopathy in
mink, chronic wasting disease in cervids, and bovine spongiform encephalopathy
(BSE) in cattle [2]. BSE was first detected in 1986 in the United Kingdom and is
the most likely cause of variant CJD (vCJD) in humans. The origin of the
original case(s) of BSE still remains an enigma. Hypotheses include (i) sheep-
or goat-derived scrapie-infected tissues included in meat and bone meal fed to
cattle, (ii) a previously undetected sporadic or genetic bovine TSE
contaminating cattle feed or (iii) origination from a human TSE through animal
feed contaminated with human remains [3]. This study will provide support to the
hypothesis that BSE originated from a previously undetected genetic bovine TSE
contaminating cattle feed in the U.K.
Results
Here we report a case of bovine BSE associated with a mutation within the
prion protein gene (Prnp) sequence, not previously described for the bovine
Prnp. The animal (called ‘‘U.S. BSE Alabama’’) was an approximately 10 year-old
red crossbred (Bos indicus6Bos taurus) hybrid beef cow from Alabama (see
Materials and Methods). The ELISA-based BSE test (see Materials and Methods) on
brainstem from this animal was repeated five times and revealed a strongly
positive reaction with mean optical density (OD) value of 2.4060.57, whereas the
OD value of bovine control obex was ,0.04.
Confirmatory BSE tests employing Western Blot (WB) and immunohistochemical
(IHC) analyses for presence of PrPd were subsequently performed (see Materials
and Methods). Immunoblots (Figure 1A) revealed (i) presence of PrPd and (ii)
intensity of reaction with antibody P4 similar as with antibody 6H4 at identical
milligram equivalent amounts. This reaction pattern was described as being
unusual or atypical for BSE [4]. Molecular weight analysis revealed
unglycosylated and monoglycosylated isoforms of PrPd migrated with an apparent
molecular weight higher than respective isoforms of classical or C-type BSE
isolates (Figure 1A). A similar migration pattern was observed for the U.S. BSE
2004 isolate, an H-type BSE isolate [4]. Obvious lesions of spongiform
encephalopathy diagnostic for BSE were not present in the brainstem, however it
was positive for the presence of PrPd by IHC (Figure 1B). Distribution of PrPd
in the brainstem of this animal was not as uniform or as intense as seen with
the C-type U.S. BSE case from 2003 (Figure 1C) [4]. We concluded from these
studies, that this animal contracted an H-type BSE phenotype. Recent work on the
molecular characterization of cattle PrPd has allowed to define criteria for the
identification of atypical BSE cases in cattle, which showed molecular features
of the PrPd distinct from the majority of cattle with C-type BSE [4– 6]. There
have been two molecular types of unusual BSE isolates described in the
literature [5,6]: (i) a type with a lower molecular mass of the unglycosylated
isoform (L-type) and (ii) a type with higher molecular mass of the
unglycosylated isoform (H-type) when compared to C-type of BSE.
In order to confirm the specimen from this case was of cattle origin and to
determine whether the case was associated with a Prnp mutation, the full coding
sequence from exon 3 of the Prnp was amplified from DNA isolated from fresh
brainstem material (see Materials and Methods) and aligned with known Prnp
sequences from cattle, sheep and cervids (Figure 2A,B). The Prnp DNA sequence of
this animal is of bovine origin, different from sheep and cervid Prnp sequences
(Figure 2A). The sequence was heterozygous on two positions: a synonymous
polymorphism on codon 78 (CAA/CAG; Q78Q) and a non-synonymous polymorphism on
codon 211 (GAA/AAA; E211K; Figure 2A,B; Table 1). The animal had six copies of
the octapeptide repeat region on both of its Prnp alleles (Figure 2A,B). The
finding of the E211K mutation is of significant interest because an identical
mutation, E200K, at the homologous codon 200 position in human Prnp (Figure 1D)
has been described as the most common mutation in humans with genetic CJD
[7].
Discussion
Our results demonstrate for the first time a potential pathogenic mutation
(E211K) within the Prnp gene of a bovine with an H-type BSE phenotype at a
position representing the most common mutation in humans (E200K) associated with
genetic TSEs [7]. This mutation was not found in the Prnp gene of other North
American (1 H-type U.S.; 1 H-type and 1 L-type Canadian) and European (7 H-type
and 3 L-type cases) cattle [8] and a miniature zebu (H-type) [9] with atypical
BSE phenotypes. The functional significance of this finding, however, remains
unknown. Importantly, the penetrance of the E200K mutation in humans is very
high [7,10]. The origin of atypical BSE cases still remains unexplained. Several
hypotheses have been considered including the existence of a previously
unrecognized ‘‘sporadic’’ form of a TSE in this species. The detection of the
E211K Prnp mutation, known to be pathogenic in humans, in a 10 year old hybrid
cow (Bos indicus6Bos taurus) with H-type BSE could provide additional support to
the following hypotheses: (i) that U.K. BSE has been acquired from a genetic
case or cases of cattle BSE, (ii) that all three etiological forms of human TSEs
(sporadic, genetic and infectious) are also present in cattle, and (iii) that
BSE started on the Indian subcontinent. However, more data are required to
support these hypotheses. It is well known, that large amounts of mammalian
protein material were imported from India to the U.K. during the relevant time
period (late 1970s and early 1980s) [3]. Therefore it could be speculated that
one possible route of contamination of U.K. cattle with BSE was through animal
feed containing imported meat and bone meal material contaminated with a case or
cases of genetic BSE.
Epidemiological investigations conducted by USDA personnel failed to reveal
any evidence of a feed source contaminated with TSE material fed to this animal
(
http://www.aphis.usda.gov/newsroom/
hot_issues/bse/downloads/EPI_Final5-2-06.pdf). There are several possibilities
for the origin of the Prnp K211 allele in animal B14842: (i) it arose de novo in
a germ line cell from the U.S. BSE Alabama animal or one of its parents; (ii) it
arose as a somatic mutation in the U.S. BSE Alabama animal (rather unlikely),
and (iii) it is present in a cattle population or breed yet to be found.
Recently, it was determined that the 2-year-old heifer offspring of the U.S. BSE
Alabama cow also carries the E211K polymorphism, indicating that the allele is
heritable and may persist within the cattle population (11) In a recent
epidemiological study which included 6062 cattle from 5 commercial beef
processing plants (3892 carcasses) and 2170 registered cattle from 42 breeds.,
the K211 allele was not detected using a newly developed mass spectrometry assay
specific for the E211K variant [12]. These data indicate a rather low prevalence
of the E211K variant of less than 1 in 2000 cattle when using Bayesian analysis
[12]. This newly developed assay system for K211 [12] will offer the possibility
for genetic surveillance of cattle for rare pathogenic mutations that may be
associated with BSE.
Materials and Methods
Animals
The animal B14842 (called U.S. BSE Alabama case), approximately 10 years
old as determined by dentition, was a red crossbred (Bos indicus6Bos taurus)
hybrid beef cow found in lateral recumbency on a farm in Alabama. She was
euthanized by the attending veterinarian, the brainstem removed for BSE testing
and shipped to the Georgia Veterinary Medical Diagnostic Laboratory, where it
was found to be positive by a rapid enzyme-linked immunosorbent assay
(ELISA)-based BSE test. The brainstem was then forwarded to the National
Veterinary Services Laboratories in Ames, IA, USA, for confirmatory testing. The
classical BSE case was reported previously [4].
ELISA test
The rapid BSE test used in the U.S. is the Platelia/TeSeETM ELISA BSE test
(Bio-Rad, Hercules, CA, USA). Fresh samples from the brainstem were used for the
analysis and the samples were treated with proteinase K (PK) in order to digest
the PKsensitive normal prion protein, PrPc.
Imunoblot Analysis
Brain homogenates from the U.S. BSE Alabama case (animal B14842) were
prepared from 1.1 gram of brainstem material and analyzed using the
PrionicsH-Check Western Kit (Prionics, Schlieren, Switzerland) with
modifications and the OIE-recommended Scrapie Associated Fibril (SAF)-Immunoblot
method (
http://www.
oie.int/eng/normes/mmanual/A_summry.htm). Preparation and analysis of brainstem
homogenate using both methods has been described previously [4]. Please note
that the brain homogenates were treated with PK (2 U/ml) for 60 minutes at 37uC
before Western Blot analysis in order to digest the PK-sensitive PrPc. As
positive control samples, BSE-positive brain material from the U.S. BSE cases
2003 and 2004 as well as a sheep scrapie isolate were used. As negative
controls, brain material from a BSE-negative cow was used.
Immunohistochemistry
Brain tissue was placed in 10% buffered formalin and after a minimum of 4
days of fixation appropriate sections of brainstem in the obex region were put
in cassettes and kept in fresh formalin until they were processed for routine
paraffin embedding. The procedure was described in detail previously [4]. The
IHC results were interpreted as follows: (i) positive for PrPd: pink to red and
(ii) background and negative for PrPd: only blue background. As positive
controls, slides from the brainstem of a BSE-positive cow, obtained from the
United Kingdom and from the U.S. BSE Case 2003 were used. As negative controls,
slides from brainstem material of BSE-negative cattle and scrapie-negative sheep
were used.
DNA isolation, PCR amplification and sequence analysis Genomic DNA was
extracted from 200 ml of a 10% brain homogenate as described previously [4]. The
fragment was sequenced in duplicate using the original two primers and two
internal primers (available upon request) for a total of 8 reactions. Databases
were searched using standard nucleotide-nucleotide BLAST at the National Center
for Biotechnology Information Web Site (
http://www.ncbi.nlm.nih.gov). The
database is a collection of sequences from several sources, including GenBank
and Reference Sequence. The nucleotide sequences of the Prnp gene of the U.S.
BSE Alabama case was aligned using both CLUSTAL V and CLUSTAL W with the
following GENBANK accession numbers: AY335912 (bovine), AY367641 (bovine),
AF016227(elk), AY275712 (white-tailed deer), AF166334 (sheep), AJ567986 (sheep),
and the Canadian BSE case [12] using Lasergene version 5.07 software
(DNASTAR-Madison WI). Accession number
The GenBank accession number for the Prnp gene of U.S. BSE Alabama case is
EU809428.
Acknowledgments
The authors would like to thank D. Clouser and K. Hassall for their
excellent technical support and Dr. M. Kehrli for his encouragement and support.
We thank Drs. T. Baron, S. Czub and T. Seuberlich for sharing DNA from atypical
BSE cases. The authors also wish to thank the seven State/University Veterinary
Diagnostic Laboratories for their efforts in screening U.S. cattle for the
presence of BSE that enabled detection of this new allele.
Mention of trade names or commercial products in this article is solely for
the purpose of providing specific information and does not imply recommendation
or endorsement by the U.S. Department of Agriculture. Author Contributions
Conceived and designed the experiments: JAR SMH. Performed the experiments:
JAR SMH. Analyzed the data: JAR SMH. Contributed reagents/materials/analysis
tools: JAR. Wrote the paper: JAR SMH. References
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2. Wells GA, Wilesmith JW (1995) The neuropathology and epidemiology of
bovine spongiform encephalopathy. Brain Pathol 1995 5: 91–103.
3. Colchester AC, Colchester NT (2005) The origin of bovine spongiform
encephalopathy: the human prion disease hypothesis. Lancet 366: 856–61.
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Identification and characterization of two bovine spongiform encephalopathy
cases diagnosed in the United States. J Vet Diagn Invest 19: 142–154.
5. Jacobs JG, Langeveld JP, Biacabe AG, Acutis PL, Polak MP, et al. (2007)
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from a geographical region spanning a wide area in Europe. J Clin Microbiol 45:
1821–1829.
6. Baron T, Biacabe AG, Arsac JN, Benestad S, Groschup MH (2007) Atypical
transmissible spongiform encephalopathies (TSEs) in ruminants. Vaccine 25:
5625–5630.
7. Kovacs GG, Puopolo M, Ladogana A, Pocchiari M, Budka H, et al. (2005)
Genetic prion disease: the EUROCJD experience. Hum Genet 118: 166–174.
8. Clawson ML, Richt JA, Baron T, Biacabe AG, Czub S, et al. (2008)
Association of a bovine prion gene haplotype with atypical BSE. PLoS ONE 3:
e1830. doi:10.1371/journal.pone.0001830.
9. Seuberlich T, Botteron C, Wenker C, Cafe´-Marcal VA, Oevermann A, et al.
(2006) Spongiform encephalopathy in a miniature zebu. Emerg Infect Dis 12:
1950–1953.
10. Kong Q, Surewicz WK, Peterson RB, Zou W, Chen SG, et al. (2004)
Inherited Prion Diseases. In: Prusiner SB, ed. Prion Biology and Diseases. New
York: Cold Spring Harbor Monograph Series 41. 1050 p.
11. Nicholson EN, Brunelle BW, Richt JA, Kehrli Jr ME, Greenlee JJ (2008)
Identification of a heritable polymorphism in bovine PRNP associated with
genetic transmissible spongiform encephalopathy: evidence of heritable BSE. PLoS
ONE 3: e2912. doi:10.1371/journal.pone.0002912.
12. Heaton MP, Keele JW, Harhay GP, Richt JA, Koohmaraie M, et al. (2008)
Prevalence of the prion gene E211K variant in U.S. cattle. BMC Vet Res 4:
25.
13. Coulthart MB, Mogk R, Rancourt JM, Godal DL, Czub S (2003) Prion
protein gene sequence of Canada’s first non-imported case of bovine spongiform
encephalopathy (BSE). Genome 46: 1005–1009.
Table 1. DNA sequence analysis of codon 211 of the bovine Prnp and codon
200 of the human Prnp. Nucleotide Amino Acid Bovine majority G A A/G A A
E211/E211 U.S. 2006 BSE case G A A/A A A E211/K211 Human majority G A G/G A G
E200/E200 Human genetic CJD G A G/A A G E200/K200
doi:10.1371/journal.ppat.1000156.t001 Novel Bovine Prion Protein Gene Mutation
PLoS
Figure 1. Analysis of brainstem samples from BSE-infected animals employing
various methods. (A) Hybrid Immunoblot Analysis using enriched samples: Lanes
1–5: monoclonal antibody 6H4 (raised against human PrP residues 144–152), lanes
8–12 monoclonal antibody P4 (raised against ovine PrP residues 89–104): 1 =
sheep scrapie control, 2 mg; 2 = classical BSE (2003 U.S. BSE case), 2 mg; 3 =
H-type BSE case (2004 U.S. BSE case), 2 mg; 4 = U.S. BSE Alabama case, 1 mg; 5 =
U.S. BSE Alabama case, 2.5 mg; 6,7 = protein weight maker; 8 = U.S. BSE Alabama
case, 2.5 mg; 9 = U.S. BSE Alabama case, 1 mg; 10 = H-type BSE case (2004 U.S.
BSE case), 2 mg; 11 = classical BSE (2003 U.S. BSE case), 2 mg; 12 = sheep
scrapie control, 2 mg. (B) Immunohistochemistry of the U.S. BSE Alabama case
(H-type BSE) using PrP-specific monoclonal antibody F99/97.6.1. Brainstem at the
level of obex was examined. Bar = 35 mm. (C) Immunohistochemistry of a classical
BSE case [4] using PrP-specific monoclonal antibody F99/97.6.1. Brainstem at the
level of obex was examined. Spongiform changes are found in the area with highly
PrPd-positive cells. Bar = 90 mm. doi:10.1371/journal.ppat.1000156.g001
Figure 2. Alignment of bovine, ovine, cervid and human Prnp sequences. (A)
Nucleotide sequences. Standard single letter codes are used for nucleotides. Y =
C or T; R = A or G; K =G or T; W= A or T. Boxed area indicates the 6th
octapeptide-repeat of the bovine protein (U.S. BSE Alabama case and sequence
AY335912). Additional Prnp sequences are as follows: AJ567986 (sheep), AF016227
(elk), Hsap M13899 (human, normal) and Hsap PRNPvar [human, variant; see [13]].
(B) Amino acid sequences. Standard IUPAC single letter codes are used for amino
acids. Codon numbering refers to the most common six-copy octapeptide repeat
allele for Bos Taurus. Boxed area indicates the 6th octapeptide repeat of the
bovine protein [animals B14842 [4] and AY335912]. AJ567986 (sheep), AF016227
(elk), Hsap M13899 (human, normal) and Hsap PRNPvar [human, variant; see [13]]
each contain a 5 octapeptide repeat region in the protein.
doi:10.1371/journal.ppat.1000156.g002
2015
(c) The commonest form of CJD occurs as a sporadic disease, the cause of
which is unknown, although genetic factors (particularly the codon 129
polymorphism in the prion protein gene (PRNP)) influence disease susceptibility.
The familial forms of human TSEs (see Box 1) appear to have a solely genetic
origin and are closely associated with mutations or insertions in the PRNP gene.
Most, but not all, of the familial forms of human TSEs have been transmitted
experimentally to animals. *** There are no known familial or genetic TSEs of
animals, although polymorphisms in the PRNP gene of some species (sheep for
example) may influence the length of the incubation period and occurrence of
disease.
just saying, now let’s look at banned suspect mad cow in commerce in
Alabama feed, as source for Alabama atypical gh type BSE, some 9 years post
ruminant feed ban ;
BANNED MAD COW FEED IN COMMERCE IN ALABAMA
Date: September 6, 2006 at 7:58 am PST PRODUCT
a) EVSRC Custom dairy feed, Recall # V-130-6;
b) Performance Chick Starter, Recall # V-131-6;
c) Performance Quail Grower, Recall # V-132-6;
d) Performance Pheasant Finisher, Recall # V-133-6.
CODE None RECALLING FIRM/MANUFACTURER Donaldson & Hasenbein/dba J&R
Feed Service, Inc., Cullman, AL, by telephone on June 23, 2006 and by letter
dated July 19, 2006. Firm initiated recall is complete.
REASON
Dairy and poultry feeds were possibly contaminated with ruminant based
protein.
VOLUME OF PRODUCT IN COMMERCE 477.72 tons
DISTRIBUTION AL
______________________________
PRODUCT Bulk custom dairy pre-mixes,
Recall # V-120-6 CODE None RECALLING FIRM/MANUFACTURER Ware Milling Inc.,
Houston, MS, by telephone on June 23, 2006. Firm initiated recall is complete.
REASON Possible contamination of dairy animal feeds with ruminant derived meat
and bone meal.
VOLUME OF PRODUCT IN COMMERCE 350 tons
DISTRIBUTION AL and MS
______________________________
PRODUCT
a) Tucker Milling, LLC Tm 32% Sinking Fish Grower, #2680-Pellet, 50 lb.
bags, Recall # V-121-6;
b) Tucker Milling, LLC #31120, Game Bird Breeder Pellet, 50 lb. bags,
Recall # V-122-6;
c) Tucker Milling, LLC #31232 Game Bird Grower, 50 lb. bags, Recall #
V-123-6;
d) Tucker Milling, LLC 31227-Crumble, Game Bird Starter, BMD Medicated, 50
lb bags, Recall # V-124-6;
e) Tucker Milling, LLC #31120, Game Bird Breeder, 50 lb bags, Recall #
V-125-6;
f) Tucker Milling, LLC #30230, 30 % Turkey Starter, 50 lb bags, Recall #
V-126-6;
g) Tucker Milling, LLC #30116, TM Broiler Finisher, 50 lb bags, Recall #
V-127-6
CODE All products manufactured from 02/01/2005 until 06/20/2006 RECALLING
FIRM/MANUFACTURER Recalling Firm: Tucker Milling LLC, Guntersville, AL, by
telephone and visit on June 20, 2006, and by letter on June 23, 2006.
Manufacturer: H. J. Baker and Brothers Inc., Stamford, CT. Firm initiated recall
is ongoing.
REASON Poultry and fish feeds which were possibly contaminated with
ruminant based protein were not labeled as "Do not feed to ruminants".
VOLUME OF PRODUCT IN COMMERCE 7,541-50 lb bags
DISTRIBUTION AL, GA, MS, and TN
END OF ENFORCEMENT REPORT FOR AUGUST 9, 2006
###
Subject: MAD COW FEED RECALL AL AND FL VOLUME OF PRODUCT IN COMMERCE 125
TONS Products manufactured from 02/01/2005 until 06/06/2006
Date: August 6, 2006 at 6:16 pm PST PRODUCT
a) CO-OP 32% Sinking Catfish, Recall # V-100-6;
b) Performance Sheep Pell W/Decox/A/N, medicated, net wt. 50 lbs, Recall #
V-101-6;
c) Pro 40% Swine Conc Meal -- 50 lb, Recall # V-102-6;
d) CO-OP 32% Sinking Catfish Food Medicated, Recall # V-103-6;
e) "Big Jim's" BBB Deer Ration, Big Buck Blend, Recall # V-104-6;
f) CO-OP 40% Hog Supplement Medicated Pelleted, Tylosin 100 grams/ton, 50
lb. bag, Recall # V-105-6;
g) Pig Starter Pell II, 18% W/MCDX Medicated 282020, Carbadox -- 0.0055%,
Recall # V-106-6;
h) CO-OP STARTER-GROWER CRUMBLES, Complete Feed for Chickens from Hatch to
20 Weeks, Medicated, Bacitracin Methylene Disalicylate, 25 and 50 Lbs, Recall #
V-107-6;
i) CO-OP LAYING PELLETS, Complete Feed for Laying Chickens, Recall #
108-6;
j) CO-OP LAYING CRUMBLES, Recall # V-109-6;
k) CO-OP QUAIL FLIGHT CONDITIONER MEDICATED, net wt 50 Lbs, Recall #
V-110-6;
l) CO-OP QUAIL STARTER MEDICATED, Net Wt. 50 Lbs, Recall # V-111-6;
m) CO-OP QUAIL GROWER MEDICATED, 50 Lbs, Recall # V-112-6 CODE
Product manufactured from 02/01/2005 until 06/06/2006
RECALLING FIRM/MANUFACTURER Alabama Farmers Cooperative, Inc., Decatur, AL,
by telephone, fax, email and visit on June 9, 2006. FDA initiated recall is
complete.
REASON Animal and fish feeds which were possibly contaminated with ruminant
based protein not labeled as "Do not feed to ruminants".
VOLUME OF PRODUCT IN COMMERCE 125 tons
DISTRIBUTION AL and FL
END OF ENFORCEMENT REPORT FOR AUGUST 2, 2006
###
MAD COW FEED RECALL USA EQUALS 10,878.06 TONS NATIONWIDE Sun Jul 16, 2006
09:22 71.248.128.67
RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINE -- CLASS II
______________________________
PRODUCT
a) PRO-LAK, bulk weight, Protein Concentrate for Lactating Dairy Animals,
Recall # V-079-6;
b) ProAmino II, FOR PREFRESH AND LACTATING COWS, net weight 50lb (22.6 kg),
Recall # V-080-6;
c) PRO-PAK, MARINE & ANIMAL PROTEIN CONCENTRATE FOR USE IN ANIMAL FEED,
Recall # V-081-6;
d) Feather Meal, Recall # V-082-6 CODE
a) Bulk
b) None
c) Bulk
d) Bulk
RECALLING FIRM/MANUFACTURER H. J. Baker & Bro., Inc., Albertville, AL,
by telephone on June 15, 2006 and by press release on June 16, 2006. Firm
initiated recall is ongoing.
REASON
Possible contamination of animal feeds with ruminent derived meat and bone
meal.
VOLUME OF PRODUCT IN COMMERCE 10,878.06 tons
DISTRIBUTION Nationwide
END OF ENFORCEMENT REPORT FOR July 12, 2006
###
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
Tuesday, March 2, 2010
Animal Proteins Prohibited in Ruminant Feed/Adulterated/Misbranded Rangen
Inc 2/11/10 USA
Monday, March 1, 2010
ANIMAL PROTEIN I.E. MAD COW FEED IN COMMERCE A REVIEW 2010
Terry S. Singeltary Sr. (Submitted question): Monday, April 5, 2010
Update on Feed Enforcement Activities to Limit the Spread of BSE April 5,
2010
Friday, April 23, 2010
Upcoming BSE Webinar on Thursday, April 22, 2010 a review
P.9.21
Molecular characterization of BSE in Canada
Jianmin Yang1, Sandor Dudas2, Catherine Graham2, Markus Czub3, Tim
McAllister1, Stefanie Czub1 1Agriculture and Agri-Food Canada Research Centre,
Canada; 2National and OIE BSE Reference Laboratory, Canada; 3University of
Calgary, Canada
Background: Three BSE types (classical and two atypical) have been
identified on the basis of molecular characteristics of the misfolded protein
associated with the disease. To date, each of these three types have been
detected in Canadian cattle.
Objectives: This study was conducted to further characterize the 16
Canadian BSE cases based on the biochemical properties of there associated
PrPres. Methods: Immuno-reactivity, molecular weight, glycoform profiles and
relative proteinase K sensitivity of the PrPres from each of the 16 confirmed
Canadian BSE cases was determined using modified Western blot analysis.
Results: Fourteen of the 16 Canadian BSE cases were C type, 1 was H type
and 1 was L type. The Canadian H and L-type BSE cases exhibited size shifts and
changes in glycosylation similar to other atypical BSE cases. PK digestion under
mild and stringent conditions revealed a reduced protease resistance of the
atypical cases compared to the C-type cases. N terminal- specific antibodies
bound to PrPres from H type but not from C or L type. The C-terminal-specific
antibodies resulted in a shift in the glycoform profile and detected a fourth
band in the Canadian H-type BSE.
Discussion: The C, L and H type BSE cases in Canada exhibit molecular
characteristics similar to those described for classical and atypical BSE cases
from Europe and Japan. This supports the theory that the importation of BSE
contaminated feedstuff is the source of C-type BSE in Canada. It also suggests a
similar cause or source for atypical BSE in these countries.
P.5.21
Parallels between different forms of sheep scrapie and types of
Creutzfeldt-Jakob disease (CJD)
Wiebke M. Wemheuer1, Sylvie L. Benestad2, Arne Wrede1, Wilhelm E.
Wemheuer3, Tatjana Pfander1, Bjørn Bratberg2, Bertram Brenig3,Walter J.
Schulz-Schaeffer1 1University Medical Center Goettingen, Germany; 2Institute of
Veterinary Medicine Oslo, Norway; 3Institute of Veterinary Medicine Goettingen,
Germany
Background: Scrapie in sheep and goats is often regarded as the archetype
of prion diseases. In 1998, a new form of scrapie – atypical/Nor98 scrapie – was
described that differed from classical scrapie in terms of epidemiology, Western
blot profile, the distribution of pathological prion protein (PrPSc) in the body
and its stability against proteinase K. In a similar way, distinct disease types
exist in sporadic Creutzfeldt-Jakob disease (CJD). They differ with regard to
their clinical outcome, Western blot profile and PrPSc deposition pattern in the
central nervous system (CNS). Objectives: The comparison of PrPSc deposits in
sheep scrapie and human sporadic CJD.
Methods: Tissues of the CNS of sheep with classical scrapie, sheep with
atypical/Nor98 scrapie and 20 patients with sporadic CJD were examined using the
sensitive Paraffin Embedded Tissue (PET) blot method. The results were compared
with those obtained by immunohistochemistry. With the objective of gaining
information on the protein conformation, the PrPSc of classical and
atypical/Nor98 sheep scrapie and sporadic CJD was tested for its stability
against denaturation with guanidine hydrochloride (GdnHCl) using a Membrane
Adsorption Assay.
Results: The PrPSc of atypical/Nor98 scrapie cases and of CJD prion type 1
patients exhibits a mainly reticular/synaptic deposition pattern in the brain
and is relatively sensitive to denaturation with GdnHCl. In contrast classical
scrapie cases and CJD prion type 2 patients have a more complex PrPSc deposition
pattern in common that consists of larger PrPSc aggregates and the PrPSc itself
is comparatively stable against denaturation.
Discussion: The similarity between CJD types and scrapie types indicates
that at least two comparable forms of the misfolded prion protein exist beyond
species barriers and can elicit prion diseases. It seems therefore reasonable to
classify classical and atypical/Nor98 scrapie – in analogy to the existing CJD
types – as different scrapie types.
Monday, December 14, 2009
Similarities between Forms of Sheep Scrapie and Creutzfeldt-Jakob Disease
Are Encoded by Distinct Prion Types
Monday, December 14, 2009
Similarities between Forms of Sheep Scrapie and Creutzfeldt-Jakob Disease
Are Encoded by Distinct Prion Types
hmmm, this is getting interesting now...
> Sporadic CJD type 1 and atypical/ Nor98 scrapie are characterized by
fine (reticular) deposits,
see also ;
> All of the Heidenhain variants were of the methionine/ methionine type
1 molecular subtype.
see full text ;
Monday, December 14, 2009
Similarities between Forms of Sheep Scrapie and Creutzfeldt-Jakob Disease
Are Encoded by Distinct Prion Types
Epidemiology of Scrapie in the United States 1977
Tuesday, April 28, 2009
Nor98-like Scrapie in the United States of America
P03.141
Aspects of the Cerebellar Neuropathology in Nor98
Gavier-Widén, D1; Benestad, SL2; Ottander, L1; Westergren, E1 1National
Veterinary Insitute, Sweden; 2National Veterinary Institute,
Norway Nor98 is a prion disease of old sheep and goats. This atypical form
of scrapie was first described in Norway in 1998. Several features of Nor98 were
shown to be different from classical scrapie including the distribution of
disease associated prion protein (PrPd) accumulation in the brain. The
cerebellum is generally the most affected brain area in Nor98. The study here
presented aimed at adding information on the neuropathology in the cerebellum of
Nor98 naturally affected sheep of various genotypes in Sweden and Norway. A
panel of histochemical and immunohistochemical (IHC) stainings such as IHC for
PrPd, synaptophysin, glial fibrillary acidic protein, amyloid, and cell markers
for phagocytic cells were conducted. The type of histological lesions and tissue
reactions were evaluated. The types of PrPd deposition were characterized. The
cerebellar cortex was regularly affected, even though there was a variation in
the severity of the lesions from case to case. Neuropil vacuolation was more
marked in the molecular layer, but affected also the granular cell layer. There
was a loss of granule cells. Punctate deposition of PrPd was characteristic. It
was morphologically and in distribution identical with that of synaptophysin,
suggesting that PrPd accumulates in the synaptic structures. PrPd was also
observed in the granule cell layer and in the white matter. The pathology
features of Nor98 in the cerebellum of the affected sheep showed similarities
with those of sporadic Creutzfeldt-Jakob disease in humans.
***The pathology features of Nor98 in the cerebellum of the affected sheep
showed similarities with those of sporadic Creutzfeldt-Jakob disease in
humans.
PR-26
NOR98 SHOWS MOLECULAR FEATURES REMINISCENT OF GSS
R. Nonno1, E. Esposito1, G. Vaccari1, E. Bandino2, M. Conte1, B.
Chiappini1, S. Marcon1, M. Di Bari1, S.L. Benestad3, U. Agrimi1 1 Istituto
Superiore di Sanità, Department of Food Safety and Veterinary Public Health,
Rome, Italy (romolo.nonno@iss.it); 2 Istituto Zooprofilattico della Sardegna,
Sassari, Italy; 3 National Veterinary Institute, Department of Pathology, Oslo,
Norway
Molecular variants of PrPSc are being increasingly investigated in sheep
scrapie and are generally referred to as "atypical" scrapie, as opposed to
"classical scrapie". Among the atypical group, Nor98 seems to be the best
identified. We studied the molecular properties of Italian and Norwegian Nor98
samples by WB analysis of brain homogenates, either untreated, digested with
different concentrations of proteinase K, or subjected to enzymatic
deglycosylation. The identity of PrP fragments was inferred by means of
antibodies spanning the full PrP sequence. We found that undigested brain
homogenates contain a Nor98-specific PrP fragment migrating at 11 kDa (PrP11),
truncated at both the C-terminus and the N-terminus, and not N-glycosylated.
After mild PK digestion, Nor98 displayed full-length PrP (FL-PrP) and
N-glycosylated C-terminal fragments (CTF), along with increased levels of PrP11.
Proteinase K digestion curves (0,006-6,4 mg/ml) showed that FL-PrP and CTF are
mainly digested above 0,01 mg/ml, while PrP11 is not entirely digested even at
the highest concentrations, similarly to PrP27-30 associated with classical
scrapie. Above 0,2 mg/ml PK, most Nor98 samples showed only PrP11 and a fragment
of 17 kDa with the same properties of PrP11, that was tentatively identified as
a dimer of PrP11. Detergent solubility studies showed that PrP11 is insoluble in
2% sodium laurylsorcosine and is mainly produced from detergentsoluble,
full-length PrPSc. Furthermore, among Italian scrapie isolates, we found that a
sample with molecular and pathological properties consistent with Nor98 showed
plaque-like deposits of PrPSc in the thalamus when the brain was analysed by
PrPSc immunohistochemistry. Taken together, our results show that the
distinctive pathological feature of Nor98 is a PrP fragment spanning amino acids
~ 90-155. This fragment is produced by successive N-terminal and C-terminal
cleavages from a full-length and largely detergent-soluble PrPSc, is produced in
vivo and is extremely resistant to PK digestion.
*** Intriguingly, these conclusions suggest that some pathological features
of Nor98 are reminiscent of Gerstmann-Sträussler-Scheinker disease.
119
A newly identified type of scrapie agent can naturally infect sheep with
resistant PrP genotypes
Annick Le Dur*,?, Vincent Béringue*,?, Olivier Andréoletti?, Fabienne
Reine*, Thanh Lan Laï*, Thierry Baron§, Bjørn Bratberg¶, Jean-Luc Vilotte?,
Pierre Sarradin**, Sylvie L. Benestad¶, and Hubert Laude*,?? +Author
Affiliations
*Virologie Immunologie Moléculaires and ?Génétique Biochimique et
Cytogénétique, Institut National de la Recherche Agronomique, 78350
Jouy-en-Josas, France; ?Unité Mixte de Recherche, Institut National de la
Recherche Agronomique-Ecole Nationale Vétérinaire de Toulouse, Interactions Hôte
Agent Pathogène, 31066 Toulouse, France; §Agence Française de Sécurité Sanitaire
des Aliments, Unité Agents Transmissibles Non Conventionnels, 69364 Lyon,
France; **Pathologie Infectieuse et Immunologie, Institut National de la
Recherche Agronomique, 37380 Nouzilly, France; and ¶Department of Pathology,
National Veterinary Institute, 0033 Oslo, Norway
***Edited by Stanley B. Prusiner, University of California, San Francisco,
CA (received for review March 21, 2005)
Abstract Scrapie in small ruminants belongs to transmissible spongiform
encephalopathies (TSEs), or prion diseases, a family of fatal neurodegenerative
disorders that affect humans and animals and can transmit within and between
species by ingestion or inoculation. Conversion of the host-encoded prion
protein (PrP), normal cellular PrP (PrPc), into a misfolded form, abnormal PrP
(PrPSc), plays a key role in TSE transmission and pathogenesis. The intensified
surveillance of scrapie in the European Union, together with the improvement of
PrPSc detection techniques, has led to the discovery of a growing number of
so-called atypical scrapie cases. These include clinical Nor98 cases first
identified in Norwegian sheep on the basis of unusual pathological and PrPSc
molecular features and "cases" that produced discordant responses in the rapid
tests currently applied to the large-scale random screening of slaughtered or
fallen animals. Worryingly, a substantial proportion of such cases involved
sheep with PrP genotypes known until now to confer natural resistance to
conventional scrapie. Here we report that both Nor98 and discordant cases,
including three sheep homozygous for the resistant PrPARR allele (A136R154R171),
efficiently transmitted the disease to transgenic mice expressing ovine PrP, and
that they shared unique biological and biochemical features upon propagation in
mice. *** These observations support the view that a truly infectious TSE agent,
unrecognized until recently, infects sheep and goat flocks and may have
important implications in terms of scrapie control and public health.
Monday, December 1, 2008
When Atypical Scrapie cross species barriers
Authors
Andreoletti O., Herva M. H., Cassard H., Espinosa J. C., Lacroux C., Simon
S., Padilla D., Benestad S. L., Lantier F., Schelcher F., Grassi J., Torres, J.
M., UMR INRA ENVT 1225, Ecole Nationale Veterinaire de Toulouse.France;
ICISA-INlA, Madrid, Spain; CEA, IBiTec-5, DSV, CEA/Saclay, Gif sur Yvette cedex,
France; National Veterinary Institute, Postboks 750 Sentrum, 0106 Oslo, Norway,
INRA IASP, Centre INRA de Tours, 3738O Nouzilly, France.
Content
Atypical scrapie is a TSE occurring in small ruminants and harbouring
peculiar clinical, epidemiological and biochemical properties. Currently this
form of disease is identified in a large number of countries. In this study we
report the transmission of an atypical scrapie isolate through different species
barriers as modeled by transgenic mice (Tg) expressing different species PRP
sequence.
The donor isolate was collected in 1995 in a French commercial sheep flock.
inoculation into AHQ/AHQ sheep induced a disease which had all
neuro-pathological and biochemical characteristics of atypical scrapie.
Transmitted into Transgenic mice expressing either ovine or PrPc, the isolate
retained all the described characteristics of atypical scrapie.
Surprisingly the TSE agent characteristics were dramatically different
v/hen passaged into Tg bovine mice. The recovered TSE agent had biological and
biochemical characteristics similar to those of atypical BSE L in the same mouse
model. Moreover, whereas no other TSE agent than BSE were shown to transmit into
Tg porcine mice, atypical scrapie was able to develop into this model, albeit
with low attack rate on first passage.
Furthermore, after adaptation in the porcine mouse model this prion showed
similar biological and biochemical characteristics than BSE adapted to this
porcine mouse model. Altogether these data indicate.
(i) the unsuspected potential abilities of atypical scrapie to cross
species barriers
(ii) the possible capacity of this agent to acquire new characteristics
when crossing species barrier
These findings raise some interrogation on the concept of TSE strain and on
the origin of the diversity of the TSE agents and could have consequences on
field TSE control measures.
P.4.23
Transmission of atypical BSE in humanized mouse models
Liuting Qing1, Wenquan Zou1, Cristina Casalone2, Martin Groschup3, Miroslaw
Polak4, Maria Caramelli2, Pierluigi Gambetti1, Juergen Richt5, Qingzhong Kong1
1Case Western Reserve University, USA; 2Instituto Zooprofilattico Sperimentale,
Italy; 3Friedrich-Loeffler-Institut, Germany; 4National Veterinary Research
Institute, Poland; 5Kansas State University (Previously at USDA National Animal
Disease Center), USA
Background: Classical BSE is a world-wide prion disease in cattle, and the
classical BSE strain (BSE-C) has led to over 200 cases of clinical human
infection (variant CJD). Atypical BSE cases have been discovered in three
continents since 2004; they include the L-type (also named BASE), the H-type,
and the first reported case of naturally occurring BSE with mutated bovine PRNP
(termed BSE-M). The public health risks posed by atypical BSE were largely
undefined.
Objectives: To investigate these atypical BSE types in terms of their
transmissibility and phenotypes in humanized mice. Methods: Transgenic mice
expressing human PrP were inoculated with several classical (C-type) and
atypical (L-, H-, or Mtype) BSE isolates, and the transmission rate, incubation
time, characteristics and distribution of PrPSc, symptoms, and histopathology
were or will be examined and compared.
Results: Sixty percent of BASE-inoculated humanized mice became infected
with minimal spongiosis and an average incubation time of 20-22 months, whereas
only one of the C-type BSE-inoculated mice developed prion disease after more
than 2 years. Protease-resistant PrPSc in BASE-infected humanized Tg mouse
brains was biochemically different from bovine BASE or sCJD. PrPSc was also
detected in the spleen of 22% of BASE-infected humanized mice, but not in those
infected with sCJD. Secondary transmission of BASE in the humanized mice led to
a small reduction in incubation time. The atypical BSE-H strain is also
transmissible with distinct phenotypes in the humanized mice, but no BSE-M
transmission has been observed so far.
Discussion: Our results demonstrate that BASE is more virulent than
classical BSE, has a lymphotropic phenotype, and displays a modest transmission
barrier in our humanized mice.
BSE-H is also transmissible in our humanized Tg mice.
The possibility of more than two atypical BSE strains will be
discussed.
Supported by NINDS NS052319, NIA AG14359, and NIH AI 77774.
O.4.4
PrPSc distribution pattern in cattle experimentally challenged with H-type
and L-type atypical BSE
Anne Buschmann1, Ute Ziegler1, Leila McIntyre2, Markus Keller1, Ron
Rogers3, Bob Hills3, Martin H. Groschup1 1Friedrich-Loeffler-Institut, INEID,
Germany; 2Faculty of Veterinary Medicine, University of Calgary, Canada; 3Health
Canada, Ottawa, Canada
Background: After the detection of two novel BSE forms designated H-type
and L-type BSE, the question of the pathogenesis and the agent distribution in
cattle affected with these forms was fully open. From initial studies, it was
already known that the PrPSc distribution in L-type BSE affected cattle differed
from that known for classical BSE (C-type) where the obex region always displays
the highest PrPSc concentrations. In contrast in L-type BSE cases, the thalamus
and frontal cortex regions showed the highest levels of the pathological prion
protein, while the obex region was only weakly involved. No information was
available on the distribution pattern in H-type BSE.
Objectives: To analyse the PrPSc and infectivity distribution in cattle
experimentally challenged with H-type and L-type BSE.
Methods: We analysed CNS and peripheral tissue samples collected from
cattle that were intracranially challenged with Htype (five animals) and L-type
(six animals) using a commercial BSE rapid test (IDEXX HerdChek),
immunohistochemistry (IHC) and a highly sensitive Western blot protocol
including a phosphotungstic acid precipitation of PrPSc (PTA-WB). Samples
collected during the preclinical and the clinical stages of the disease were
examined. For the detection of BSE infectivity, selected samples were also
inoculated into highly sensitive Tgbov XV mice overexpressing bovine prion
protein (PrPC).
Results: Analysis of a collection of fifty samples from the peripheral
nervous, lymphoreticular, digestive, reproductive, respiratory and
musculo-skeletal systems by PTA-WB, IDEXXHerdChek BSE EIA and IHC revealed a
general restriction of the PrPSc accumulation to the central nervous
system.
Discussion: Our results on the PrPSc distribution in peripheral tissues of
cattle affected with H-type and L-type BSE are generally in accordance with what
has been known for C-type BSE. Bioassays are ongoing in highly sensitive
transgenic mice in order to reveal infectivity.
O.11.3
Infectivity in skeletal muscle of BASE-infected cattle
Silvia Suardi1, Chiara Vimercati1, Fabio Moda1, Ruggerone Margherita1,
Ilaria Campagnani1, Guerino Lombardi2, Daniela Gelmetti2, Martin H. Groschup3,
Anne Buschmann3, Cristina Casalone4, Maria Caramelli4, Salvatore Monaco5,
Gianluigi Zanusso5, Fabrizio Tagliavini1 1Carlo Besta” Neurological
Institute,Italy; 2IZS Brescia, Italy; 33FLI Insel Riems, D, Germany; 4CEA-IZS
Torino, Italy; 5University of Verona, Italy
Background: BASE is an atypical form of bovine spongiform encephalopathy
caused by a prion strain distinct from that of BSE. Upon experimental
transmission to cattle, BASE induces a previously unrecognized disease phenotype
marked by mental dullness and progressive atrophy of hind limb musculature.
Whether affected muscles contain infectivity is unknown. This is a critical
issue since the BASE strain is readily transmissible to a variety of hosts
including primates, suggesting that humans may be susceptible.
Objectives: To investigate the distribution of infectivity in peripheral
tissues of cattle experimentally infected with BASE. Methods: Groups of Tg mice
expressing bovine PrP (Tgbov XV, n= 7-15/group) were inoculated both i.c. and
i.p. with 10% homogenates of a variety of tissues including brain, spleen,
cervical lymph node, kidney and skeletal muscle (m. longissimus dorsi) from
cattle intracerebrally infected with BASE. No PrPres was detectable in the
peripheral tissues used for inoculation either by immunohistochemistry or
Western blot.
Results: Mice inoculated with BASE-brain homogenates showed clinical signs
of disease with incubation and survival times of 175±15 and 207±12 days. Five
out of seven mice challenged with skeletal muscle developed a similar
neurological disorder, with incubation and survival times of 380±11 and 410±12
days. At present (700 days after inoculation) mice challenged with the other
peripheral tissues are still healthy. The neuropathological phenotype and PrPres
type of the affected mice inoculated either with brain or muscle were
indistinguishable and matched those of Tgbov XV mice infected with natural
BASE.
Discussion: Our data indicate that the skeletal muscle of cattle
experimentally infected with BASE contains significant amount of infectivity, at
variance with BSE-affected cattle, raising the issue of intraspecies
transmission and the potential risk for humans. Experiments are in progress to
assess the presence of infectivity in skeletal muscles of natural BASE.
P.5.3
Differences in the expression levels of selected genes in the brain tissue
of cattle naturally infected with classical and atypical BSE.
Magdalena Larska1, Miroslaw P. Polak1, Jan F. Zmudzinski1, Juan M. Torres2
1National Veterinary Institute, Poland; 2CISA/INIA
Background: Recently cases of BSE in older cattle named BSE type L and type
H were distinguished on the basis of atypical glycoprofiles of PrPres. The
nature of those strains is still not fully understood but it is suspected that
the atypical BSE cases are sporadic. Hitherto most BSE cases were studied in
respect to the features of PrPSc. Here we propose gene expression profiling as a
method to characterize and distinguish BSE strains.
Objectives: The aim of the study was to compare the activities of some
factors which are known to play a role in TSE’s pathogenesis in order to
distinguish the differences/similarities between all BSE types.
Methods: 10 % homogenate of brain stem tissue collected from obex region of
medulla oblongata from 20 naturally infected BSE cows (8 assigned as classical
BSE, other 8 and 4 infected with atypical BSE L type and H type respectively)
was used in the study. As negative control animals we’ve used 8 animals in the
age between 2.5 and 13 years. The genes were relatively quantified using SYBR
Green real time RT-PCR. Raw data of Ct values was transformed into normalized
relative quantities using Qbase Plus®. Results and
Discussion: In most of the tested genes significant differences in the
expression levels between the brain stem of healthy cattle and animals infected
with different BSE types were observed. In c-type BSE in comparison to healthy
and atypical BSE the overexpression of the gene of bcl-2, caspase 3, 14-3-3 and
tylosine kinase Fyn was significant.
Simultaneously in atypical BSEs type-L and type-H the levels of prion
protein, Bax and LPR gene was elevated in comparison to c-BSE. Additionally
L-BSE was characterized by the overexpression of STI1 and SOD genes compared to
the other of BSE types. The downregulation of the gene encoding NCAM1 was
observed in all BSE types in comparison to healthy cows. Different gene
expression profiles of bovine brains infected with classical and atypical BSE
indicates possible different pathogenesis or source of the disease.
O.10.1
Transmission of uncommon forms of bovine prions to transgenic mice
expressing human PrP: questions and progress
Vincent Béringue, Hubert Laude INRA, UR 892, Virologie Immunologie
Moléculaires, France
The active, large-scale testing of livestock nervous tissues for the
presence of protease-resistant prion protein (PrPres) has led to the recognition
of 2 uncommon PrPres molecular signatures, termed H-type and L-type BSE. Their
experimental transmission to various transgenic and inbred mouse lines
unambiguously demonstrated the infectious nature of such cases and the existence
of distinct prion strains in cattle. Like the classical BSE agent, H- and L-type
(or BASE) prions can propagate in heterologous species. In addition L-type
prions acquire molecular and neuropathologic phenotypic traits undistinguishable
from BSE or BSE-related agents upon transmission to transgenic mice expressing
ovine PrP (VRQ allele) or wild-type mice. An understanding of the transmission
properties of these newly recognized prions when confronted with human PrP
sequence was therefore needed. Toward this end, we inoculated mice expressing
human PrP Met129 with several field isolates. Unlike classical BSE agent, L-type
prions appeared to propagate in these mice with no obvious transmission barrier.
In contrast, we repeatedly failed to infect them with Htype prions. Ongoing
investigations aim to extend the knowledge on these uncommon strains: are these
agents able to colonize lymphoid tissue, a potential key factor for successful
transmission by peripheral route; is there any relationship between these
assumedly sporadic forms of TSE in cattle and some sporadic forms of human CJD
are among the issues that need to be addressed for a careful assessment of the
risk for cattle-to-human transmission of H- and L-type prions.
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.
P2-110
TRANSMISSION OF ATYPICAL BOVINE SPONGIFORM ENCEPHALOPATHY TO MICROCEBUS
MURINUS, A NON-HUMAN PRIMATE. DEVELOPMENT OF CLINICAL SYMPTOMS AND TISSUE
DISTRIBUTION OF PRPRES
Nadine Mestre-Frances1, Anne-Gaelle Biacabe2, Sylvie Rouland1, Thierry
Baron2, Jean-Michel Verdier1, 1INSERM U710, Montpellier, France; 2AFSSA, Lyon,
France. Contact e-mail: nfrances@univmontp2. fr
Background: Atypical BSE cases have been observed in Europe, Japan and
North America. They differ in their PrPres profiles from those found in
classical BSE. These atypical cases fall into 2 types, depending on the
molecular mass of the unglycosylated PrPres band observed by Western blot: the
L-type (lower molecular mass than the typical BSE cases) and H-type (higher
molecular mass than the typical BSE cases).
Methods: Height animals (4 males and 4 females) were intracerebrally
inoculated with 50 l of a 10% brain homogenates of atypical (L and H-type)
French BSE cases.
Results: Only one of the four lemurs challenge with H-type BSE died without
clinical signs after 19 months post inoculation (mpi), the 4 animals inoculated
with L-type BSE died at 19 mpi (2 males) and 22 mpi (2 females). Three months
before their sacrifice, they developed blindness, tremor, abnormal posture,
incoordinated movements, balance loss. Symptoms get worse according to the
disease progression, until severe ataxia. The brain tissue were biochemically
and immunocytochemically investigated for PrPres. For the H-types, spongiform
changes without PrPres accumulation were observed in the brainstem. Western blot
analysis confirmed that no PrPres was detected into the brain. For the L-types,
severe spongiosis was evidenced into the thalamus, the striatum, the
mesencephalon, and the brainstem, whereas into the cortex the spongiosis was
evidenced, but the vacuolisation was weaker. Strong deposits of PrPres was
detected by western blot, PET-blot and immunocytochemistry in the CNS: dense
accumulation was observed into the thalamus, the striatum, and the hippocampus
whereas in the cerebral cortex, PrPres was prominently accumulated in plaques.
Western blot analysis confirmed the presence of protease-resistant prion
protein.
Conclusions: L-type infected lemurs showed survival times considerably
shorter than for classical BSE strain, indicating that the disease is caused by
a very virulent distinct prion strain.
Atypical BSE (BASE) Transmitted from Asymptomatic Aging Cattle to a
Primate
Emmanuel E. Comoy1*, Cristina Casalone2, Nathalie Lescoutra-Etchegaray1,
Gianluigi Zanusso3, Sophie Freire1, Dominique Marcé1, Frédéric Auvré1,
Marie-Magdeleine Ruchoux1, Sergio Ferrari3, Salvatore Monaco3, Nicole Salès4,
Maria Caramelli2, Philippe Leboulch1,5, Paul Brown1, Corinne I. Lasmézas4,
Jean-Philippe Deslys1
1 Institute of Emerging Diseases and Innovative Therapies, CEA,
Fontenay-aux-Roses, France, 2 Istituto Zooprofilattico Sperimentale del
Piemonte, Turin, Italy, 3 Policlinico G.B. Rossi, Verona, Italy, 4 Scripps
Florida, Jupiter, Florida, United States of America, 5 Genetics Division,
Brigham & Women's Hospital, Harvard Medical School, Boston, Massachusetts,
United States of America
Abstract Top Background Human variant Creutzfeldt-Jakob Disease (vCJD)
results from foodborne transmission of prions from slaughtered cattle with
classical Bovine Spongiform Encephalopathy (cBSE). Atypical forms of BSE, which
remain mostly asymptomatic in aging cattle, were recently identified at
slaughterhouses throughout Europe and North America, raising a question about
human susceptibility to these new prion strains.
Methodology/Principal Findings Brain homogenates from cattle with classical
BSE and atypical (BASE) infections were inoculated intracerebrally into
cynomolgus monkeys (Macacca fascicularis), a non-human primate model previously
demonstrated to be susceptible to the original strain of cBSE. The resulting
diseases were compared in terms of clinical signs, histology and biochemistry of
the abnormal prion protein (PrPres). The single monkey infected with BASE had a
shorter survival, and a different clinical evolution, histopathology, and prion
protein (PrPres) pattern than was observed for either classical BSE or
vCJD-inoculated animals. Also, the biochemical signature of PrPres in the
BASE-inoculated animal was found to have a higher proteinase K sensitivity of
the octa-repeat region. We found the same biochemical signature in three of four
human patients with sporadic CJD and an MM type 2 PrP genotype who lived in the
same country as the infected bovine.
Conclusion/Significance Our results point to a possibly higher degree of
pathogenicity of BASE than classical BSE in primates and also raise a question
about a possible link to one uncommon subset of cases of apparently sporadic
CJD. Thus, despite the waning epidemic of classical BSE, the occurrence of
atypical strains should temper the urge to relax measures currently in place to
protect public health from accidental contamination by BSE-contaminated
products.
Citation: Comoy EE, Casalone C, Lescoutra-Etchegaray N, Zanusso G, Freire
S, et al. (2008) Atypical BSE (BASE) Transmitted from Asymptomatic Aging Cattle
to a Primate. PLoS ONE 3(8): e3017. doi:10.1371/journal.pone.0003017
Editor: Neil Mabbott, University of Edinburgh, United Kingdom
Received: April 24, 2008; Accepted: August 1, 2008; Published: August 20,
2008
Copyright: © 2008 Comoy et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited.
Funding: This work has been supported by the Network of Excellence
NeuroPrion.
Competing interests: CEA owns a patent covering the BSE diagnostic tests
commercialized by the company Bio-Rad.
* E-mail:
mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000110/!x-usc:mailto:emmanuel.comoy@cea.fr
Saturday, December 01, 2007
Phenotypic Similarity of Transmissible Mink Encephalopathy in Cattle and
L-type Bovine Spongiform Encephalopathy in a Mouse Model
Volume 13, Number 12–December 2007 Research
Phenotypic Similarity of Transmissible Mink Encephalopathy in Cattle and
L-type Bovine Spongiform Encephalopathy in a Mouse Model
Thierry Baron,* Anna Bencsik,* Anne-Gaëlle Biacabe,* Eric Morignat,*
andRichard A. Bessen†*Agence Française de Sécurité Sanitaire des Aliments–Lyon,
Lyon, France; and†Montana State University, Bozeman, Montana, USA
Abstract
Transmissible mink encepholapathy (TME) is a foodborne transmissible
spongiform encephalopathy (TSE) of ranch-raised mink; infection with a ruminant
TSE has been proposed as the cause, but the precise origin of TME is unknown. To
compare the phenotypes of each TSE, bovine-passaged TME isolate and 3 distinct
natural bovine spongiform encephalopathy (BSE) agents (typical BSE, H-type BSE,
and L-type BSE) were inoculated into an ovine transgenic mouse line (TgOvPrP4).
Transgenic mice were susceptible to infection with bovine-passaged TME, typical
BSE, and L-type BSE but not to H-type BSE. Based on survival periods, brain
lesions profiles, disease-associated prion protein brain distribution, and
biochemical properties of protease-resistant prion protein, typical BSE had a
distint phenotype in ovine transgenic mice compared to L-type BSE and bovine
TME.The similar phenotypic properties of L-type BSE and bovine TME in TgOvPrP4
mice suggest that L-type BSE is a much more likely candidate for the origin of
TME than is typical BSE.
snip...
Conclusion
These studies provide experimental evidence that the Stetsonville TME agent
is distinct from typical BSE but has phenotypic similarities to L-type BSE in
TgOvPrP4 mice. Our conclusion is that L-type BSE is a more likely candidate for
a bovine source of TME infection than typical BSE. In the scenario that a
ruminant TSE is the source for TME infection in mink, this would be a second
example of transmission of a TSE from ruminants to non-ruminants under natural
conditions or farming practices in addition to transmission of typical BSE to
humans, domestic cats, and exotic zoo animals(37). The potential importance of
this finding is relevant to L-type BSE, which based on experimental transmission
into humanized PrP transgenic mice and macaques, suggests that L-type BSE is
more pathogenic for humans than typical BSE (24,38).
Saturday, February 28, 2009NEW RESULTS ON IDIOPATHIC BRAINSTEM NEURONAL
CHROMATOLYSIS TYPE BSE
"All of the 15 cattle tested showed that the brains had abnormally
accumulated PrP" 2009 SEAC 102/2
Wednesday, October 08, 2008
Idiopathic Brainstem Neuronal Chromatolysis (IBNC): a novel prion protein
related disorder of cattle?
>>>Conclusions: L-type infected lemurs showed survival times
considerably shorter than for classical BSE strain, indicating that the disease
is caused by a very virulent distinct prion strain. >>>
seems the survival time was the same for the h-type BSE and the l-type BSE
i.e. 19 months post inoculation (mpi), interesting. ...TSS
Wednesday, March 31, 2010
Atypical BSE in Cattle / position: Post Doctoral Fellow
Wednesday, February 24, 2010
Transmissible Spongiform encephalopathy (TSE) animal and human TSE in North
America 14th
ICID International Scientific Exchange Brochure -
National Prion Disease Pathology Surveillance Center Cases Examined1 (July
31, 2010)
(please see video at the bottom of this url...tss)
Tuesday, August 03, 2010
Variably protease-sensitive prionopathy: A new sporadic disease of the
prion protein
Monday, August 9, 2010
Variably protease-sensitive prionopathy: A new sporadic disease of the
prion protein or just more Prionbaloney ?
Wednesday, July 28, 2010
re-Freedom of Information Act Project Number 3625-32000-086-05, Study of
Atypical BSE UPDATE July 28, 2010
Wednesday, July 28, 2010
Atypical prion proteins and IBNC in cattle DEFRA project code SE1796 FOIA
Final report
Thursday, June 24, 2010
Accumulation of L-type Bovine Prions in Peripheral Nerve Tissues
Volume 16, Number 7–July 2010
To date the OIE/WAHO assumes that the human and animal health standards set
out in the BSE chapter for classical BSE (C-Type) applies to all forms of BSE
which include the H-type and L-type atypical forms. This assumption is
scientifically not completely justified and accumulating evidence suggests that
this may in fact not be the case. Molecular characterization and the spatial
distribution pattern of histopathologic lesions and immunohistochemistry (IHC)
signals are used to identify and characterize atypical BSE. Both the L-type and
H-type atypical cases display significant differences in the conformation and
spatial accumulation of the disease associated prion protein (PrPSc) in brains
of afflicted cattle. Transmission studies in bovine transgenic and wild type
mouse models support that the atypical BSE types might be unique strains because
they have different incubation times and lesion profiles when compared to C-type
BSE. When L-type BSE was inoculated into ovine transgenic mice and Syrian
hamster the resulting molecular fingerprint had changed, either in the first or
a subsequent passage, from L-type into C-type BSE. In addition, non-human
primates are specifically susceptible for atypical BSE as demonstrated by an
approximately 50% shortened incubation time for L-type BSE as compared to
C-type. Considering the current scientific information available, it cannot be
assumed that these different BSE types pose the same human health risks as
C-type BSE or that these risks are mitigated by the same protective
measures.
Wednesday, March 31, 2010
Atypical BSE in Cattle
Let's look at some sound science on atypical Nor-98 Scrapie, shall we
;
[Although atypical scrapie is not yet ruled out, it is important to realize
this is a type of scrapie that thus far has only tended to appear as a sporadic
condition in older animals. Currently it has not been shown to follow the same
genetic tendencies for propagation as the usual scrapie.
However, the atypical phenotypic appearance has been shown to be preserved
on experimental passage.
Atypical scrapie was first identified in Norwegian sheep in 1998 and has
subsequently been identified in many countries, as Australia may join that list.
It is likely that this case will be sent to the UK for definitive
conformation.
[Ref: M Simmons, T Konold, L Thurston, et al. BMC Veterinary Research 2010,
6:14 [provisional abstract available at ]
"Background ----------- "Retrospective studies have identified cases
predating the initial identification of this form of scrapie, and
epidemiological studies have indicated that it does not conform to the behaviour
of an infectious disease, giving rise to the hypothesis that it represents
spontaneous disease. However, atypical scrapie isolates have been shown to be
infectious experimentally, through intracerebral inoculation in transgenic mice
and sheep. [Many of the neurological diseases can be transmitted by
intracerebral inoculation, which causes this moderator to approach intracerebral
studies as a tool for study, but not necessarily as a direct indication of
transmissibility of natural diseases. - Mod.TG]
"The 1st successful challenge of a sheep with 'field' atypical scrapie from
an homologous donor sheep was reported in 2007.
"Results -------- "This study demonstrates that atypical scrapie has
distinct clinical, pathological, and biochemical characteristics which are
maintained on transmission and sub-passage, and which are distinct from other
strains of transmissible spongiform encephalopathies in the same host
genotype.
"Conclusions ------------ Atypical scrapie is consistently transmissible
within AHQ homozygous sheep, and the disease phenotype is preserved on
sub-passage."
Lastly, this moderator wishes to thank Terry Singletary for some of his
behind the scenes work of providing citations and references for this posting. -
Mod.TG]
Saturday, June 5, 2010
Research Project: Transmissible Spongiform Encephalopathies: Identification
of atypical scrapie in Canadian sheep
Sunday, April 18, 2010
SCRAPIE AND ATYPICAL SCRAPIE TRANSMISSION STUDIES A REVIEW 2010
If the scrapie agent is generated from ovine DNA and thence causes disease
in other species, then perhaps, bearing in mind the possible role of scrapie in
CJD of humans (Davinpour et al, 1985), scrapie and not BSE should be the
notifiable disease. ...
Tuesday, July 27, 2010
Spontaneous generation of mammalian prions
Thursday, August 12, 2010
Seven main threats for the future linked to prions
2015
(c) The commonest form of CJD occurs as a sporadic disease, the cause of
which is unknown, although genetic factors (particularly the codon 129
polymorphism in the prion protein gene (PRNP)) influence disease susceptibility.
The familial forms of human TSEs (see Box 1) appear to have a solely genetic
origin and are closely associated with mutations or insertions in the PRNP gene.
Most, but not all, of the familial forms of human TSEs have been transmitted
experimentally to animals. *** There are no known familial or genetic TSEs of
animals, although polymorphisms in the PRNP gene of some species (sheep for
example) may influence the length of the incubation period and occurrence of
disease.
31 Jan 2015 at 20:14 GMT
*** Ruminant feed ban for cervids in the United States? ***
31 Jan 2015 at 20:14 GMT
spontaneous atypical BSE ???
don’t let anyone fool you. spontaneous TSE prion disease is a hoax in
natural cases, never proven.
all one has to do is look at France. France is having one hell of an
epidemic of atypical BSE, probably why they stopped testing for BSE, problem
solved $$$ same as the USA, that’s why they stopped testing for BSE mad cow
disease in numbers they could find any with, after those atypical BSE cases
started showing up. shut down the testing to numbers set up by OIE that are so
low, you could only by accident find a case of BSE aka mad cow disease. and this
brilliant idea by the WHO et al, to change the name of mad cow disease, thinking
that might change things is preposterous. it’s all about money now folks, when
the OIE, USDA and everyone else went along and made the TSE prion disease aka
mad cow type disease a legal trading commodity by the BSE MRR policy, I would
say everyone bit off more then they can chew, and they will just have to digest
those TSE Prions coming from North America, and like it, and just prey you don’t
get a mad cow type disease i.e. Transmissible Spongiform Encephalopathy TSE
prion disease in the decades to come, and or pass it to some other poor soul via
the iatrogenic medical surgical tissue friendly fire mode of transmission i.e.
second hand transmission. it’s real folks, just not documented much, due to lack
of trace back efforts. all iatrogenic cjd is, is sporadic cjd, until the
iatrogenic event is tracked down and documented, and put into the academic and
public domain, which very seldom happens. ...
As of December 2011, around 60 atypical BSE cases have currently been
reported in 13 countries, *** with over one third in France.
FRANCE STOPS TESTING FOR MAD COW DISEASE BSE, and here’s why, to many
spontaneous events of mad cow disease $$$
so 20 cases of atypical BSE in France, compared to the remaining 40 cases
in the remaining 12 Countries, divided by the remaining 12 Countries, about 3+
cases per country, besides Frances 20 cases. you cannot explain this away with
any spontaneous BSe. ...TSS
Sunday, October 5, 2014
France stops BSE testing for Mad Cow Disease
*** Singeltary reply ; Molecular, Biochemical and Genetic Characteristics
of BSE in Canada Singeltary reply ;
Saturday, May 09, 2015
Expression of genes involved in the T cell signalling pathway in
circulating immune cells of cattle 24 months following oral challenge with
Bovine Amyloidotic Spongiform Encephalopathy (BASE)
Sunday, May 3, 2015
PRION2015 FORT COLLINS
Sunday, March 29, 2015
Uncommon prion disease induced in macaque ten years after scrapie
inoculation
Friday, January 30, 2015
*** Scrapie: a particularly persistent pathogen ***
Friday, February 20, 2015
APHIS Freedom of Information Act (FOIA) Appeal Mouse Bio-Assays
2007-00030-A Sheep Imported From Belgium and the Presence of TSE Prion Disease
Kevin Shea to Singeltary 2015
Tuesday, December 16, 2014
Evidence for zoonotic potential of ovine scrapie prions
Hervé Cassard,1, n1 Juan-Maria Torres,2, n1 Caroline Lacroux,1, Jean-Yves
Douet,1, Sylvie L. Benestad,3, Frédéric Lantier,4, Séverine Lugan,1, Isabelle
Lantier,4, Pierrette Costes,1, Naima Aron,1, Fabienne Reine,5, Laetitia
Herzog,5, Juan-Carlos Espinosa,2, Vincent Beringue5, & Olivier
Andréoletti1, Affiliations Contributions Corresponding author Journal name:
Nature Communications Volume: 5, Article number: 5821 DOI:
doi:10.1038/ncomms6821 Received 07 August 2014 Accepted 10 November 2014
Published 16 December 2014 Article tools Citation Reprints Rights &
permissions Article metrics
Abstract
Although Bovine Spongiform Encephalopathy (BSE) is the cause of variant
Creutzfeldt Jakob disease (vCJD) in humans, the zoonotic potential of scrapie
prions remains unknown. Mice genetically engineered to overexpress the human
prion protein (tgHu) have emerged as highly relevant models for gauging the
capacity of prions to transmit to humans. These models can propagate human
prions without any apparent transmission barrier and have been used used to
confirm the zoonotic ability of BSE. Here we show that a panel of sheep scrapie
prions transmit to several tgHu mice models with an efficiency comparable to
that of cattle BSE. The serial transmission of different scrapie isolates in
these mice led to the propagation of prions that are phenotypically identical to
those causing sporadic CJD (sCJD) in humans. These results demonstrate that
scrapie prions have a zoonotic potential and raise new questions about the
possible link between animal and human prions.
Subject terms: Biological sciences• Medical research At a glance
why do we not want to do TSE transmission studies on chimpanzees $
5. A positive result from a chimpanzee challenged severly would likely
create alarm in some circles even if the result could not be interpreted for
man. I have a view that all these agents could be transmitted provided a large
enough dose by appropriate routes was given and the animals kept long enough.
Until the mechanisms of the species barrier are more clearly understood it might
be best to retain that hypothesis.
snip...
R. BRADLEY
Suspect symptoms
What if you can catch old-fashioned CJD by eating meat from a sheep
infected with scrapie?
28 Mar 01 Most doctors believe that sCJD is caused by a prion protein
deforming by chance into a killer. But Singeltary thinks otherwise. He is one of
a number of campaigners who say that some sCJD, like the variant CJD related to
BSE, is caused by eating meat from infected animals. Their suspicions have
focused on sheep carrying scrapie, a BSE-like disease that is widespread in
flocks across Europe and North America.
Now scientists in France have stumbled across new evidence that adds
weight to the campaigners' fears. To their complete surprise, the researchers
found that one strain of scrapie causes the same brain damage in mice as sCJD.
"This means we cannot rule out that at least some sCJD may be caused by
some strains of scrapie," says team member Jean-Philippe Deslys of the French
Atomic Energy Commission's medical research laboratory in Fontenay-aux-Roses,
south-west of Paris. Hans Kretschmar of the University of Göttingen, who
coordinates CJD surveillance in Germany, is so concerned by the findings that he
now wants to trawl back through past sCJD cases to see if any might have been
caused by eating infected mutton or lamb...
2001
Suspect symptoms
What if you can catch old-fashioned CJD by eating meat from a sheep
infected with scrapie?
28 Mar 01
Like lambs to the slaughter
31 March 2001
by Debora MacKenzie Magazine issue 2284.
FOUR years ago, Terry Singeltary watched his mother die horribly from a
degenerative brain disease. Doctors told him it was Alzheimer's, but Singeltary
was suspicious. The diagnosis didn't fit her violent symptoms, and he demanded
an autopsy. It showed she had died of sporadic Creutzfeldt-Jakob disease.
Most doctors believe that sCJD is caused by a prion protein deforming by
chance into a killer. But Singeltary thinks otherwise. He is one of a number of
campaigners who say that some sCJD, like the variant CJD related to BSE, is
caused by eating meat from infected animals. Their suspicions have focused on
sheep carrying scrapie, a BSE-like disease that is widespread in flocks across
Europe and North America.
Now scientists in France have stumbled across new evidence that adds
weight to the campaigners' fears. To their complete surprise, the researchers
found that one strain of scrapie causes the same brain damage in mice as sCJD.
"This means we cannot rule out that at least some sCJD may be caused by
some strains of scrapie," says team member Jean-Philippe Deslys of the French
Atomic Energy Commission's medical research laboratory in Fontenay-aux-Roses,
south-west of Paris. Hans Kretschmar of the University of Göttingen, who
coordinates CJD surveillance in Germany, is so concerned by the findings that he
now wants to trawl back through past sCJD cases to see if any might have been
caused by eating infected mutton or lamb.
Scrapie has been around for centuries and until now there has been no
evidence that it poses a risk to human health. But if the French finding means
that scrapie can cause sCJD in people, countries around the world may have
overlooked a CJD crisis to rival that caused by BSE.
Deslys and colleagues were originally studying vCJD, not sCJD. They
injected the brains of macaque monkeys with brain from BSE cattle, and from
French and British vCJD patients. The brain damage and clinical symptoms in the
monkeys were the same for all three. Mice injected with the original sets of
brain tissue or with infected monkey brain also developed the same symptoms.
As a control experiment, the team also injected mice with brain tissue
from people and animals with other prion diseases: a French case of sCJD; a
French patient who caught sCJD from human-derived growth hormone; sheep with a
French strain of scrapie; and mice carrying a prion derived from an American
scrapie strain. As expected, they all affected the brain in a different way from
BSE and vCJD. But while the American strain of scrapie caused different damage
from sCJD, the French strain produced exactly the same pathology.
"The main evidence that scrapie does not affect humans has been
epidemiology," says Moira Bruce of the neuropathogenesis unit of the Institute
for Animal Health in Edinburgh, who was a member of the same team as Deslys.
"You see about the same incidence of the disease everywhere, whether or not
there are many sheep, and in countries such as New Zealand with no scrapie." In
the only previous comparisons of sCJD and scrapie in mice, Bruce found they were
dissimilar.
But there are more than 20 strains of scrapie, and six of sCJD. "You would
not necessarily see a relationship between the two with epidemiology if only
some strains affect only some people," says Deslys. Bruce is cautious about the
mouse results, but agrees they require further investigation. Other trials of
scrapie and sCJD in mice, she says, are in progress.
People can have three different genetic variations of the human prion
protein, and each type of protein can fold up two different ways. Kretschmar has
found that these six combinations correspond to six clinical types of sCJD: each
type of normal prion produces a particular pathology when it spontaneously
deforms to produce sCJD.
But if these proteins deform because of infection with a disease-causing
prion, the relationship between pathology and prion type should be different, as
it is in vCJD. "If we look at brain samples from sporadic CJD cases and find
some that do not fit the pattern," says Kretschmar, "that could mean they were
caused by infection."
There are 250 deaths per year from sCJD in the US, and a similar incidence
elsewhere. Singeltary and other US activists think that some of these people
died after eating contaminated meat or "nutritional" pills containing dried
animal brain. Governments will have a hard time facing activists like Singeltary
if it turns out that some sCJD isn't as spontaneous as doctors have insisted.
Deslys's work on macaques also provides further proof that the human
disease vCJD is caused by BSE. And the experiments showed that vCJD is much more
virulent to primates than BSE, even when injected into the bloodstream rather
than the brain. This, says Deslys, means that there is an even bigger risk than
we thought that vCJD can be passed from one patient to another through
contaminated blood transfusions and surgical instruments.
Friday, May 22, 2015
*** Chronic Wasting Disease and Program
Updates - 2014 NEUSAHA Annual Meeting 12-14 May 2014 ***
Thursday, March 20, 2014
CHRONIC WASTING DISEASE CWD TSE PRION OF CERVID AND THE POTENTIAL FOR
HUMAN TRANSMISSION THEREFROM 2014
Tuesday, July 01, 2014
*** CHRONIC WASTING DISEASE CWD TSE PRION DISEASE, GAME FARMS, AND
POTENTIAL RISK FACTORS THERE FROM ***
Friday, May 15, 2015
Grass Plants Bind, Retain, Uptake, and
Transport Infectious Prions
Report
land spreading of the TSE prion
Thursday, July 03, 2014
*** How Chronic Wasting Disease is affecting deer population and what’s
the risk to humans and pets? ***
Thursday
CWD TO HUMANS, AND RISK FACTORS THERE FROM (see latest science)
Tuesday, November 04, 2014
*** Six-year follow-up of a point-source exposure to CWD contaminated
venison in an Upstate New York community: risk behaviours and health outcomes
2005–2011
CWD TO HUMANS, AND RISK FACTORS THERE FROM (see latest science)
Monday, March 09, 2015
*** Chronic Wasting Disease CWD TSE prion and human animal risk factor
there from ***
Thursday, April 30, 2015
*** Immediate and ongoing detection of prions in the blood of hamsters and
deer following oral, nasal, or blood inoculations
Friday, January 30, 2015
*** Scrapie: a particularly persistent pathogen ***
Sunday, April 12, 2015
*** Research Project: Transmission, Differentiation, and Pathobiology of
Transmissible Spongiform Encephalopathies 2014 Annual Report ***
Saturday, April 11, 2015
*** ISU veterinary researchers study retinal scans as early detection
method for mad cow disease
Tuesday, May 19, 2015
*** COUNTRY OF ORIGIN LABELING COOL H.R. 2393 Agriculture Chairman K.
Michael Conaway (R-TX) Fears of US imports infected with mad cow disease is
emerging as an issue in trans-Pacific trade talks
Saturday, May 09, 2015
Psychiatric Symptoms in Patients With Sporadic Creutzfeldt-Jakob Disease
NORTH AMERICA is awash with the Transmissible Spongiform Encephalopathy
TSE Prion disease in many species, they are spreading, and humans and other
animals have become more and more exposed to the TSE Prion by many different
routes and sources.
CAN YOU SAY IATROGENIC !
Tuesday, May 26, 2015
*** Minimise transmission risk of CJD and vCJD in healthcare settings Last
updated 15 May 2015 ***
Tuesday, April 21, 2015
*** Transmissible Spongiform Encephalopathy Advisory Committee TSEAC
MEETING SCHEDULED FOR June 1, 2015
Sunday, April 06, 2014
SPORADIC CJD and the potential for zoonotic transmission there from,
either directly or indirectly via friendly fire iatrogenic mode, evidence to
date
Friday, January 10, 2014
*** vpspr, sgss, sffi, TSE, an iatrogenic by-product of gss, ffi, familial
type prion disease, what it ???
PRION2015 FORT COLLINS
Wednesday May 27
14:45 Jean-Phillipe Deslys Atomic Energy Commission, France,
Transmission of prions to primates after extended silent incubation
periods: * IMPLICATIONS FOR BSE AND SCRAPIE RISK ASSESSMENT IN HUMAN
POPULATIONS.
16:45
Quingzhong Kong Case Western Reserve University
Zoonotic Potential of CWD Prions
*** Kuru Video ***
Kuru: The Science and The Sorcery
*** Scrapie Video
*** Human Mad Cow Video
*** USA sporadic CJD MAD COW DISEASE HAS HUGE PROBLEM Video
2014
***Moreover, L-BSE has been transmitted more easily to transgenic mice
overexpressing a human PrP [13,14] or to primates [15,16] than C-BSE.
***It has been suggested that some sporadic CJD subtypes in humans may
result from an exposure to the L-BSE agent.
*** Lending support to this hypothesis, pathological and biochemical
similarities have been observed between L-BSE and an sCJD subtype (MV genotype
at codon 129 of PRNP) [17], and between L-BSE infected non-human primate and
another sCJD subtype (MM genotype) [15].
snip...
Monday, October 10, 2011
EFSA Journal 2011 The European Response to BSE: A Success Story
snip...
EFSA and the European Centre for Disease Prevention and Control (ECDC)
recently delivered a scientific opinion on any possible epidemiological or
molecular association between TSEs in animals and humans (EFSA Panel on
Biological Hazards (BIOHAZ) and ECDC, 2011). This opinion confirmed Classical
BSE prions as the only TSE agents demonstrated to be zoonotic so far
*** but the possibility that a small proportion of human cases so far
classified as "sporadic" CJD are of zoonotic origin could not be excluded.
*** Moreover, transmission experiments to non-human primates suggest that
some TSE agents in addition to Classical BSE prions in cattle (namely L-type
Atypical BSE, Classical BSE in sheep, transmissible mink encephalopathy (TME)
and chronic wasting disease (CWD) agents) might have zoonotic potential.
snip...
Thursday, August 12, 2010
Seven main threats for the future linked to prions
First threat
The TSE road map defining the evolution of European policy for protection
against prion diseases is based on a certain numbers of hypotheses some of which
may turn out to be erroneous. In particular, a form of BSE (called atypical
Bovine Spongiform Encephalopathy), recently identified by systematic testing in
aged cattle without clinical signs, may be the origin of classical BSE and thus
potentially constitute a reservoir, which may be impossible to eradicate if a
sporadic origin is confirmed.
*** Also, a link is suspected between atypical BSE and some apparently
sporadic cases of Creutzfeldt-Jakob disease in humans.
*** These atypical BSE cases constitute an unforeseen first threat that
could sharply modify the European approach to prion diseases.
Second threat
snip...
Sunday, November 23, 2014
*** Confirmed Variant Creutzfeldt-Jakob Disease (variant CJD) Case in Texas
in June 2014 confirmed as USA case NOT European ***
Monday, November 3, 2014
USA CJD TSE PRION UNIT, TEXAS, SURVEILLANCE UPDATE NOVEMBER 2014
National Prion Disease Pathology Surveillance Center Cases Examined1
(October 7, 2014)
***6 Includes 11 cases in which the diagnosis is pending, and 19
inconclusive cases;
***7 Includes 12 (11 from 2014) cases with type determination pending in
which the diagnosis of vCJD has been excluded.
***The sporadic cases include 2660 cases of sporadic Creutzfeldt-Jakob
disease (sCJD),
***50 cases of Variably Protease-Sensitive Prionopathy (VPSPr)
***and 21 cases of sporadic Fatal Insomnia (sFI).
Thursday, January 15, 2015
41-year-old Navy Commander with sporadic Creutzfeldt–Jakob disease CJD TSE
Prion: Case Report
Subject: *** Becky Lockhart 46, Utah’s first female House speaker, dies
diagnosed with the extremely rare Creutzfeldt-Jakob disease aka mad cow type
disease
what is CJD ? just ask USDA inc., and the OIE, they are still feeding the
public and the media industry fed junk science that is 30 years old.
why doesn’t some of you try reading the facts, instead of rubber stamping
everything the USDA inc says.
sporadic CJD has now been linked to BSE aka mad cow disease, Scrapie, and
there is much concern now for CWD and risk factor for humans.
My sincere condolences to the family and friends of the House Speaker Becky
Lockhart. I am deeply saddened hear this.
with that said, with great respect, I must ask each and every one of you
Politicians that are so deeply saddened to hear of this needless death of the
Honorable House Speaker Becky Lockhart, really, cry me a friggen river. I am
seriously going to ask you all this...I have been diplomatic for about 17 years
and it has got no where. people are still dying. so, are you all stupid or
what??? how many more need to die ??? how much is global trade of beef and other
meat products that are not tested for the TSE prion disease, how much and how
many bodies is this market worth?
Saturday, January 17, 2015
*** Becky Lockhart 46, Utah’s first female House speaker, dies diagnosed
with the extremely rare Creutzfeldt-Jakob disease
*** ALERT new variant Creutzfeldt Jakob Disease nvCJD or vCJD, sporadic CJD
strains, TSE prion aka Mad Cow Disease United States of America Update December
14, 2014 Report ***
*** Creutzfeldt-Jakob Disease Public Health Crisis VIDEO
PLEASE REMEMBER ;
The Akron, Ohio-based CJD Foundation said the Center for Disease Control
revised that number in October of 2004 to about one in 9,000 CJD cases per year
in the population group age 55 and older.
HAVE YOU GOT YOUR CJD QUESTIONNAIRE ASKING REAL QUESTIONS PERTAINING TO
ROUTE AND SOURCE OF THE TSE AGENT THAT KILLED YOUR LOVED ONE ???
if not, why not...
Friday, November 30, 2007
CJD QUESTIONNAIRE USA CWRU AND CJD FOUNDATION
Self-Propagative Replication of Ab Oligomers Suggests Potential
Transmissibility in Alzheimer Disease
Received July 24, 2014; Accepted September 16, 2014; Published November 3,
2014
Singeltary comment ;
Saturday, December 13, 2014
Terry S. Singeltary Sr. Publications TSE prion disease
Diagnosis and Reporting of Creutzfeldt-Jakob Disease
Singeltary, Sr et al. JAMA.2001; 285: 733-734. Vol. 285 No. 6, February 14,
2001 JAMA
snip...
Terry S. Singeltary Sr. P.O. Box 42 Bacliff, Texas USA 77518
