----- Original Message -----
From: Terry S. Singeltary Sr.
To: [email protected]. ; [email protected].
Sent: Thursday, August 25, 2005 9:16 AM
Subject: Importation of Whole Cuts of Boneless Beef from Japan [Docket No. 05-004-1] RIN 0579-AB93 TSS SUBMISSION
Greetings Dr. Colgrove and Miss Johnson,
Thank you for taking this submission via email. i have had trouble submitting via the comment page due to the length of my submission. I was not sure that my file attachment that i submitted via the ;
EDOCKET: Go to http://www.epa.gov/feddocket
I submitted yesterday, just did not know if the file reached anyone. so to make sure, I am sending to you to submit for me.
many thanks,
Terry
From: TSS ()
Subject: Importation of Whole Cuts of Boneless Beef from Japan [Docket No. 05-004-1] RIN 0579-AB93 TSS SUBMISSION
Date: August 24, 2005 at 2:47 pm PST
August 24, 2005
Importation of Whole Cuts of Boneless Beef from Japan [Docket No. 05-004-1] RIN 0579-AB93 TSS SUBMISSION
Greetings APHIS ET AL,
My name is Terry S. Singeltary Sr.
I would kindly like to comment on [Docket No. 05-004-1] RIN 0579-AB93 ;
PROPOSED RULES
Exportation and importation of animals and animal products:
Whole cuts of boneless beef from-
Japan,
48494-48500 [05-16422]
[Federal Register: August 18, 2005 (Volume 70, Number 159)]
[Proposed Rules]
[Page 48494-48500]
From the Federal Register Online via GPO Access [wais.access.gpo.gov]
[DOCID:fr18au05-7]
========================================================================
Proposed Rules
Federal Register
________________________________________________________________________
This section of the FEDERAL REGISTER contains notices to the public of
the proposed issuance of rules and regulations. The purpose of these
notices is to give interested persons an opportunity to participate in
the rule making prior to the adoption of the final rules.
========================================================================
[[Page 48494]]
DEPARTMENT OF AGRICULTURE
Animal and Plant Health Inspection Service
9 CFR Part 94
[Docket No. 05-004-1]
RIN 0579-AB93
Importation of Whole Cuts of Boneless Beef from Japan
AGENCY: Animal and Plant Health Inspection Service, USDA.
ACTION: Proposed rule.
-----------------------------------------------------------------------
SUMMARY: We are proposing to amend the regulations governing the
importation of meat and other edible animal products by allowing, under
certain conditions, the importation of whole cuts of boneless beef from
Japan. We are proposing this action in response to a request from the
Government of Japan and after conducting an analysis of the risk that
indicates that such beef can be safely imported from Japan under the
conditions described in this proposal.
DATES: We will consider all comments that we receive on or before
September 19, 2005.
ADDRESSES: You may submit comments by any of the following methods:
EDOCKET: Go to http://www.epa.gov/feddocket to submit or
snip...
BSE infectivity has never been demonstrated in the muscle tissue of
cattle experimentally or naturally infected with BSE at any stage of
the disease. Studies performed using TSEs other than BSE in non-bovine
animals have detected prions in muscle tissue. However, the
international scientific community largely considers that these studies
cannot be directly extrapolated to BSE in cattle because of the
significant interactions between the host species and the prion strain
involved.
Pathogenesis studies of naturally and experimentally infected
cattle have not detected BSE infectivity in blood. However,
transmission of BSE was demonstrated in sheep that received a
transfusion of a large volume of blood drawn from other sheep that were
experimentally infected with the BSE agent. The United Kingdom's
Department for Environment, Food and Rural Affairs' Spongiform
Encephalopathy Advisory Committee (SEAC) and the European Commission's
Scientific Steering Committee (SSC), which are scientific advisory
committees, evaluated the implication of this finding in relation to
food safety.\5\ The SEAC concluded that the finding did not represent
grounds for recommending any changes to the current control measures
for BSE. The SSC determined that the research results do not support
the hypothesis that bovine blood or muscle meat constitute a risk to
human health.\6\
snip...
BSE Risk Factors for Whole Cuts of Boneless Beef
The most significant risk management strategy for ensuring the
safety of whole cuts of boneless beef is the prevention of cross-
contamination of the beef with SRMs during stunning and slaughter of
the animal. Control measures that prevent contamination of such beef
involve the establishment of procedures for the removal of SRMs,
prohibitions on air-injection stunning and pithing, and splitting of
carcasses. These potential pathways for contamination and the control
measures that prevent contamination are described in detail in the risk
analysis for this rulemaking.
SRM Removal. Research has demonstrated that SRMs from infected
cattle may contain BSE infectivity. Because infectivity has not been
demonstrated in muscle tissue, the most important mitigation measure
for whole cuts of boneless beef is the careful removal and segregation
of SRMs. Removal of SRMs in a manner that avoids contamination of the
beef with SRMs minimizes the risk of exposure to materials that have
been demonstrated to contain the BSE agent in cattle.
snip...
Variant Creutzfeldt-Jakob disease (vCJD), a chronic and fatal
neurodegenerative disease of humans, has been linked since 1996 through
epidemiological, neuropathological, and experimental data to exposure
to the BSE agent, most likely through consumption of cattle products
contaminated with the agent before BSE control measures were in place.
To date, approximately 170 probable and confirmed cases of vCJD have
been identified worldwide. The majority of these cases have either been
identified in the United Kingdom or were linked to exposure that
occurred in the United Kingdom, and all cases have been linked to
exposure in countries with native cases of BSE. Some studies estimate
that more than 1 million cattle may have been infected with BSE
throughout the epidemic in the United Kingdom. This number of infected
cattle could have introduced a significant amount of infectivity into
the human food supply. Yet, the low number of cases of vCJD identified
to date indicates that there is a substantial species barrier that
protects humans from widespread illness due to exposure to the BSE
agent.
snip...
International Guidelines on BSE
International guidelines for trade in animal and animal products
are developed by the World Organization for Animal Health (formerly
known as the Office International des Epizooties (OIE)), which is
recognized by the World Trade Organization (WTO) as the international
organization responsible for the development of standards, guidelines,
and recommendations with respect to animal health and zoonoses
(diseases that are transmissible from animals to humans). The OIE
guidelines for trade in terrestrial animals (mammals, birds, and bees)
are detailed in the Terrestrial Animal Health Code (available on the
internet at http://www.oie.int). The guidelines on BSE are contained in
Chapter 2.3.13 of the Code and supplemented by Appendix 3.8.4 of the
Code.
snip...end
http://a257.g.akamaitech.net/7/257/2422/01jan20051800/edocket.access.gpo.gov/2005/05-16422.htm http://a257.g.akamaitech.net/7/257/2422/01jan20051800/edocket.access.gpo.gov/2005/pdf/05-16422.pdf
Greetings again APHIS ET AL,
THIS is not correct. IN fact, there are several factors i would like to kindly address.
Muscle tissue has recently been detected with PrPSc
in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve) of the 11th BSE
cow in Japan (Yoshifumi Iwamaru et al). also recently, Aguzzi et al Letter to the Editor
Vet Pathol 42:107-108 (2005), Prusiner et al CDI test is another example of detection
of the TSE agent in muscle in sCJD, Herbert Budka et al CJD and inclusion body myositis:
Abundant Disease-Associated Prion Protein in Muscle, and older studies from Watson
Meldrum et al Scrapie agent in muscle - Pattison I A (1990), references as follow ;
PrPSc distribution of a natural case of bovine
spongiform encephalopathy
Yoshifumi Iwamaru, Yuka Okubo, Tamako Ikeda, Hiroko Hayashi, Mori-
kazu Imamura, Takashi Yokoyama and Morikazu Shinagawa
Priori Disease Research Center, National Institute of Animal Health, 3-1-5
Kannondai, Tsukuba 305-0856 Japan [email protected]
Abstract
Bovine spongiform encephalopathy (BSE) is a disease of cattle that causes
progressive neurodegeneration of the central nervous system. Infectivity
of BSE agent is accompanied with an abnormal isoform of prion protein
(PrPSc).
The specified risk materials (SRM) are tissues potentially carrying BSE
infectivity. The following tissues are designated as SRM in Japan: the
skull including the brain and eyes but excluding the glossa and the masse-
ter muscle, the vertebral column excluding the vertebrae of the tail, spinal
cord, distal illeum. For a risk management step, the use of SRM in both
animal feed or human food has been prohibited. However, detailed
PrPSc distribution remains obscure in BSE cattle and it has caused con-
troversies about definitions of SRM. Therefore we have examined PrPSc
distribution in a BSE cattle by Western blotting to reassess definitions of
SRM.
The 11th BSE case in Japan was detected in fallen stock surveillance.
The carcass was stocked in the refrigerator. For the detection of PrPSc,
200 mg of tissue samples were homogenized. Following collagenase
treatment, samples were digested with proteinase K. After digestion,
PrPSc was precipitated by sodium phosphotungstate (PTA). The pellets
were subjected to Western blotting using the standard procedure.
Anti-prion protein monoclonal antibody (mAb) T2 conjugated horseradish
peroxidase was used for the detection of PrPSc.
PrPSc was detected in brain, spinal cord, dorsal root ganglia, trigeminal
ganglia, sublingual ganglion, retina. In addition, PrPSc was also detected
in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve).
Our results suggest that the currently accepted definitions of SRM in
BSE cattle may need to be reexamined. ...
179
T. Kitamoto (Ed.)
PRIONS
Food and Drug Safety
================
ALSO from the International Symposium of Prion Diseases held in Sendai, October 31, to
November 2, 2004;
Bovine spongiform encephalopathy (BSE) in Japan
snip...
"Furthermore, current studies into transmission of cases of BSE that are
atypical or that develop in young cattle are expected to amplify the BSE
prion"
NO. Date conf. Farm Birth place and Date Age at diagnosis
8. 2003.10.6. Fukushima Tochigi 2001.10.13. 23
9. 2003.11.4. Hiroshima Hyogo 2002.1.13. 21
Test results
# 8b, 9c cows Elisa Positive, WB Positive, IHC negative, histopathology
negative
b = atypical BSE case
c = case of BSE in a young animal
b,c, No PrPSc on IHC, and no spongiform change on histology
International Symposium of Prion Diseases held in Sendai, October 31, to
November 2, 2004.
The hardback book title is 'PRIONS' Food and Drug Safety
T. Kitamoto (Ed.)
Tetsuyuki Kitamoto
Professor and Chairman
Department of Prion Research
Tohoku University School of Medicine
2-1 SeiryoAoba-ku, Sendai 980-8575, JAPAN
TEL +81-22-717-8147 FAX +81-22-717-8148
e-mail; [email protected]
Symposium Secretariat
Kyomi Sasaki
TEL +81-22-717-8233 FAX +81-22-717-7656
e-mail: [email protected]
================================
107
Vet Pathol 42:107–108 (2005)
Letters to the Editor
Editor:
Absence of evidence is not always evidence of absence.
In the article ''Failure to detect prion protein (PrPres) by
immunohistochemistry in striated muscle tissues of animals
experimentally inoculated with agents of transmissible spongiform
encephalopathy,'' recently published in Veterinary
Pathology (41:78–81, 2004), PrPres was not detected in striated
muscle of experimentally infected elk, cattle, sheep, and
raccoons by immunohistochemistry (IHC). Negative IHC,
however, does not exclude the presence of PrPSc. For example,
PrPres was detected in skeletal muscle in 8 of 32
humans with the prion disease, sporadic Creutzfeldt-Jakob
disease (CJD), using sodium phosphotungstic acid (NaPTA)
precipitation and western blot.1 The NaPTA precipitation,
described by Wadsworth et al.,3 concentrates the abnormal
isoform of the prion, PrPres, from a large tissue homogenate
volume before western blotting. This technique has increased
the sensitivity of the western blot up to three orders
of magnitude and could be included in assays to detect
PrPres. Extremely conspicuous deposits of PrPres in muscle
were detected by IHC in a recent case report of an individual
with inclusion body myositis and CJD.2 Here, PrPres was
detected in the muscle by immunoblotting, IHC, and paraf-
fin-embedded tissue blot. We would therefore caution that,
in addition to IHC, highly sensitive biochemical assays and
bioassays of muscle are needed to assess the presence or
absence of prions from muscle in experimental and natural
TSE cases.
Christina Sigurdson, Markus Glatzel, and Adriano Aguzzi
Institute of Neuropathology
University Hospital of Zurich
Zurich, Switzerland
References
1 Glatzel M, Abela E, et al: Extraneural pathologic prion
protein in sporadic Creutzfeldt-Jakob disease. N Engl J
Med 349(19):1812–1820, 2003
2 Kovacs GG, Lindeck-Pozza E, et al: Creutzfeldt-Jakob
disease and inclusion body myositis: abundant diseaseassociated
prion protein in muscle. Ann Neurol 55(1):
121–125, 2004
3 Wadsworth JDF, Joiner S, et al: Tissue distribution of protease
resistant prion protein in variant CJD using a highly
sensitive immuno-blotting assay. Lancet 358:171–180,
2001
===================================
Corinna Kaarlela, News Director
Source: Jennifer O'Brien
[email protected]
415-476-2557
14 February 2005
Diagnosis of prions in patients should utilize novel strategy, team says
A technique for detecting prions in tissue, developed in recent years by
UCSF scientists, is significantly more sensitive than the diagnostic
procedures currently used to detect the lethal particles in samples of
brain tissue from patients, according to a study performed by a UCSF team.
The finding indicates that the diagnostic technique, known as the
conformation-dependent immunoassay (CDI), should be established as the
standard approach for brain biopsies of patients suspected of having the
disease, they say. The team is exploring whether the CDI might be
adapted to detect prions in blood and muscle.
The finding suggests that reliance on the current methods for detecting
prions in human brain tissue -- microscopic examination of tissue for
the telltale vacuoles that form in brain cells and immunohistochemistry
(IHC), which involves detecting prions in brain sections using prion
protein-specific antibodies -- may have led to an under diagnosis of the
disease in patients in recent years, they say. (A definitive diagnosis
of the disease in humans is made only on autopsy, when a
neuropathologist can analyze multiple brain regions for vacuoles and
evidence of prions by IHC, and it is estimated that only 50 percent of
human cases are autopsied, in part because many pathologists do not want
to risk infection during the autopsy.)
In the study, the team compared the ability of the CDI and the two
traditional diagnostic techniques to detect prions in various brain
samples from 28 patients diagnosed on autopsy as having one of several
human forms of the disease -- sporadic, familial or iatrogenic
Creutzfeldt-Jakob disease (CJD). While the CDI detected the biochemical
signal for prions in 100 percent of the samples studied, the traditional
tests failed to detect the prion in a high proportion of cases. For
example, in an experiment that focused on 18 brain regions from eight
patients with sporadic CJD, the CDI detected prions in 100 percent of
the samples, while IHC detected them in 22 percent and routine tissue
examination in 17 percent.
"In about 80 percent of the different brain regions examined, prions
were not consistently detected by either IHC or routine histology that
measure vacuolation. In contrast, the CDI was always positive in all
regions of the brain," says the lead author of the study, Jiri Safar,
MD, associate adjunct professor of neurology and a member of the UCSF
Institute for Neurodegenerative Diseases, which is directed by senior
author Stanley B. Prusiner, MD, UCSF professor of neurology and
biochemistry.
"These findings indicate that histology and immunohistochemistry should
no longer be used to rule out prion disease in single-site biopsy
samples," says Safar. "The superior performance of the CDI in diagnosing
prion disease suggests that the CDI be used in future diagnostic
evaluations of prion disease, particularly for single-site brain
biopsies during life"
"If the traditional techniques are used at autopsy, they must be applied
to many cortical and subcortical samples," says co-author Stephen J.
DeArmond, MD, PhD, UCSF professor of neuropathology.
Moreover, while the study examined the efficacy of the CDI in comparison
to the two techniques routinely used by neuropathologists to detect
prions in human brain tissue, previous studies at UCSF indicate that the
CDI is also significantly more sensitive than Western blot analysis, the
technology used with IHC to detect prions in brain tissue from cattle
suspected of having bovine spongiform encephalopathy (BSE). That IHC and
Western blot analysis are relatively insensitive methods, the
researchers say, supports their ongoing assertion that the CDI should
also be used to evaluate the brain tissue of cattle.
"The studies reported here are likely to change profoundly the approach
to the diagnosis of prion disease in both humans and livestock," says Safar.
More broadly, the scientists say, the high sensitivity of the CDI
suggests that CDI-like tests could also prove useful for diagnosing
other neurodegenerative diseases, such as Alzheimer's disease,
Parkinsons's disease and fronto-temporal dementias, all of which, like
prion diseases, involve various forms of protein misprocessing. These
diseases currently are diagnosed by neuropathological analysis and
immunohistochemistry.
"Whether immunohistochemistry underestimates the incidence of one or
more of these common neurodegenerative diseases is unknown, but the CDI
could shed light on these diseases," says co-author Bruce Miller, MD,
UCSF A.W. and Mary Margaret Clausen Distinguished Professor of Neurology
and director of the UCSF Memory and Aging Center.
The finding will be printed on-line and in print on March 1, 2005 in
Proceedings of the National Academy of Sciences.
The study brings into high relief the different detection strategies of
immunohistochemistry and the CDI, both of which involve revealing the
presence of prions, known as PrPsc, by applying antibodies to brain tissue.
Standard immunohistochemistry, developed in the DeArmond lab 20 years
ago, involves using an enzyme known as a protease, or a combination of
harsh acid and high temperature treatment, to destroy normal prion
protein (PrPC), which is ubiquitous in brain tissue. Once this occurs,
scientists apply fluorescently lit antibodies that react with residues
of the relatively resistant abnormal prion protein (PrPSc), thereby
highlighting it.
The limitation of this technique is that scientists have since learned
that there is a large part of the abnormal prion protein that is
protease sensitive, and that portion escapes detection by the standard
technique. Thus, this traditional method underestimates the level of
PrPSc in tissue.
The CDI addresses this limitation by revealing the region of PrPSc that
is exposed in the normal PrPC but is buried in infectious PrPSc, using
high affinity, newly generated antibodies that identify PrPSc through
the distinct shape of the molecule, independent of proteolytic
treatments. This makes it possible to detect potentially large
concentrations of protease sensitive PrPSc molecules.
Detractors would say that it is not necessary to detect the minute level
of infectious agent that the CDI is capable of revealing, as it would be
unlikely to be lethal, says Safar. But Prusiner and his colleagues
maintain that any risk is too great when it comes to having prions in
the food supply. In addition, because even low levels of prions are
extremely resistant to inactivation, they may contaminate the
environment for many years.
Prusiner won the 1997 Nobel Prize in Physiology or Medicine for
discovering that a class of neurodegenerative diseases known as
spongiform encephalopathies was caused by prions. Prion diseases develop
in humans, cattle, sheep, deer, elk and mink.
The CDI was developed by members of the Prusiner lab. The CDI
methodology has been licensed to InPro Biotechnology, Inc.
Prusiner, Safar, DeArmond and other members of the Institute for
Neurodegenerative Diseases are scientific advisors to, or own stock in,
InPro.
Other co-authors of the study were Michael D. Geschwind, Camille
Deering, Svetlana Didorenko, Mamta Sattavat, Henry Sanchesz, Ana Serban,
Kurt Giles, of UCSF, and Martin Vey, of Behring, Marburg, Germany, and
Henry Baron, of Behring, Paris.
The study was funded by the National Institutes of Health, the John
Douglas French Foundation for Alzheimer's research, the McBean
Foundation, the State of California, Alzheimer's Disease Research Center
of California and the RR00079 General Clinical Research Center.
The UCSF Institute for Neurodegenerative Diseases:
http://ind.medschool.ucsf.edu/.
FURTHER COMPARISON OF THE CDI TO THE STANDARD DIAGNOSTIC PROCEDURES,
PROVIDED BY STEPHEN J. DEARMOND, MD, PHD, UCSF PROFESSORS OF NEUROPATHOLOGY:
Explanation as to why the CDI is more sensitive than Western blot
analysis: Studies at UCSF during development of the CDI showed that CDI
could detect prions in brain homogenates at levels that fail to produce
disease in animals (bioassay for prions). Therefore, the CDI is more
sensitive than the bioassay method, which was considered to be the most
sensitive technique for detecting prions. In contrast, Western blot
analysis for prions is significantly less sensitive than the bioassay
and is, therefore, significantly less sensitive than the CDI. Currently,
the USDA uses a combination of Western blot analysis of brainstem
homogenates and immunohistochemistry of the medulla to test cattle
suspected of having bovine spongiform encephalopathy ("mad cow
disease"). The relative insensitivity of IHC and Western blot analysis,
says DeArmond, supports the UCSF scientists' ongoing assertion that the
CDI should also be used to evaluate the brain tissue of cattle.
DeArmond cites additional evidence about Western blot analysis from a
World Health Organization (WHO) study group, which compared the CDI
method with Western blots for detection of prions in sporadic and
variant CJD brains. Based on the smallest amount of prions that could
detected by the two techniques, they found that the CDI was from 1000-
to 100,000-fold more sensitive than Western blot analysis performed in
six different research laboratories (Minor et al. Standards for the
assay of Creutzfeldt-Jakob disease specimens. J. Gen. Virol. 85:
1777-1784, 2004).
Explanation as to why IHC for prions is less sensitive than the CDI: IHC
is routinely performed on formalin-fixed, paraffin-embedded samples of
brain. Formalin fixation markedly decreases the ability of antibodies to
bind to proteins in general, which greatly weakens the IHC signal for
prions (PrPSc). In contrast, homogenates for the CDI are not treated
with reagents that decrease prion antigenicity. Moreover, to concentrate
the PrPSc for measurement by the CDI, the homogenates are exposed to
phosphotungstic acid, which selectively precipitates both
protease-sensitive and protease-resistant PrPSc that comprise prions,
but not the normal prion protein conformer found in uninfected animals,
PrPC. This step results in a higher concentration of PrPSc for detection
by the CDI. Because the PrPSc was not exposed to proteases, the CDI
measures all forms of abnormally folded PrPSc molecules.
Protease-sensitive PrPSc can account for 50 percent of the total PrPSc.
For Western analysis, homogenates of brain are treated with protease to
eliminate PrPC; however, this step also eliminates protease-sensitive
PrPSc leaving only protease-resistant PrPSc for Western blot detection
and decreasing the PrPSc signal at least in half.
###
http://pub.ucsf.edu/newsservices/releases/200502147/
PNAS | March 1, 2005 | vol. 102 | no. 9 | 3501-3506
NEUROSCIENCE
Diagnosis of human prion disease
Jiri G. Safar *, , Michael D. Geschwind , , Camille Deering
*, Svetlana Didorenko *, Mamta Sattavat ¶, Henry Sanchez ¶,
Ana Serban * , Martin Vey ||, Henry Baron **, Kurt Giles *,
, Bruce L. Miller , , Stephen J. DeArmond * , ¶ and Stanley
B. Prusiner *, , ,
*Institute for Neurodegenerative Diseases, Memory and Aging
Center, and Departments of Neurology, ¶Pathology, and
Biochemistry and Biophysics, University of California, San
Francisco, CA 94143; ||ZLB Behring, 35041 Marburg, Germany;
and **ZLB Behring, 75601 Paris, France
Contributed by Stanley B. Prusiner, December 22, 2004
Abstract
With the discovery of the prion protein (PrP),
immunodiagnostic procedures were applied to diagnose
Creutzfeldt–Jakob disease (CJD). Before development of the
conformation-dependent immunoassay (CDI), all immunoassays
for the disease-causing PrP isoform (PrPSc) used limited
proteolysis to digest the precursor cellular PrP (PrPC).
Because the CDI is the only immunoassay that measures both
the protease-resistant and protease-sensitive forms of
PrPSc, we used the CDI to diagnose human prion disease. The
CDI gave a positive signal for PrPSc in all 10–24 brain
regions (100%) examined from 28 CJD patients. A subset of 18
brain regions from 8 patients with sporadic CJD (sCJD) was
examined by histology, immunohistochemistry (IHC), and the
CDI. Three of the 18 regions (17%) were consistently
positive by histology and 4 of 18 (22%) by IHC for the 8
sCJD patients. In contrast, the CDI was positive in all 18
regions (100%) for all 8 sCJD patients. In both gray and
white matter, 90% of the total PrPSc was protease-sensitive
and, thus, would have been degraded by procedures using
proteases to eliminate PrPC. Our findings argue that the CDI
should be used to establish or rule out the diagnosis of
prion disease when a small number of samples is available as
is the case with brain biopsy. Moreover, IHC should not be
used as the standard against which all other
immunodiagnostic techniques are compared because an
immunoassay, such as the CDI, is substantially more
sensitive.
snip...
Discussion
The clinical diagnosis of human prion disease is often
difficult until the patient shows profound signs of
neurologic dysfunction. It is widely accepted that the
clinical diagnosis must be provisional until a tissue
diagnosis either confirms or rules out the clinical
assessment. Before the availability of Abs to PrP, a tissue
diagnosis was generally made by histologic evaluation of
neuropil vacuolation. IHC with
anti-glial-fibrillary-acidic-protein Abs in combination with
H&E staining preceded the use of anti-PrP Ab staining.
Recently, the role of IHC in the diagnosis of scrapie in the
brains of eight clinically affected goats inoculated with
the SSBP1 prion isolate has been challenged (14). Thalamic
samples taken from seven of eight goats with scrapie were
positive for PrPSc by Western blotting but negative by IHC.
The eighth goat was negative by Western blotting and IHC.
Consistent with these findings in goats are the data
reported here, in which IHC of formalin-fixed,
paraffin-embedded human brain samples was substantially less
sensitive than the CDI.
The CDI was developed to quantify PrPSc in tissue samples
from mammals producing prions. Concerned that limited PK
digestion was hydrolyzing some or even most of the PrPSc, we
developed a CDI that does not require PK digestion. The CDI
revealed that as much as 90% of PrPSc is sPrPSc; thus, it
was being destroyed during limited proteolytic digestion
used to hydrolyze PrPC. sPrPSc comprises 80% of PrPSc in the
frontal lobe and in the white matter (Fig. 4).
The CDI detected HuPrPSc with a sensitivity comparable to
the bioassay for prion infectivity in Tg(MHu2M) mice (Fig.
1). The high sensitivity achieved by the CDI is due to
several factors (8, 10, 11, 15). First, both sPrPSc and
rPrPSc conformers are specifically precipitated by PTA
(Table 5) (8, 9). PTA has also been used to increase the
sensitivity of Western blots enabling the detection of
rPrPSc in human muscle and other peripheral tissues (16,
17). Second, a sandwich protocol was used with the
high-affinity MAR1 mAb (11) to capture HuPrPSc and
Eu-labeled 3F4 mAb to detect HuPrPSc (12). Third, the CDI
detects PrPSc by Ab-binding to native and denatured forms of
the protein and, therefore, does not depend on proteolytic
degradation of PrPC. We chose not to perform Western blots
on most of the samples used in this study because such
immunoblots require denaturation of the sample, which
eliminates measurement of the native signal corresponding to
PrPC (Table 5). Moreover, a comparison between the CDI and
Western blotting on brain samples from sCJD and variant CJD
patients showed that the CDI was 50- to 100-fold more
sensitive (15). Additionally, Western blots combined with
densitometry are linear over a 10- to 100-fold range of
concentrations, whereas the CDI is linear over a >104-fold
range. The CDI has been automated, which not only improves
accuracy and reproducibility (10) but also allows numerous
samples to be analyzed, as reported here. Western blots are
difficult to automate and are labor intensive.
Our studies show that only the CDI detected PrPSc in all
regions examined in 24 sCJD and 3 fCJD(E200K) brains (Figs.
2 and 6). Comparative analyses demonstrated that the CDI was
vastly superior to histology and IHC. When 18 regions of 8
sCJD and 2 fCJD(E200K) brains were compared, we discovered
that histology and IHC were unreliable diagnostic tools
except for samples from a few brain regions. In contrast,
the CDI was a superb diagnostic procedure because it
detected PrPSc in all 18 regions in 8 of 8 sCJD and 2 of 2
fCJD(E200K) cases (Tables 1 and 2).
Histologic changes in prion disease have been shown to
follow the accumulation of prions as measured by bioassay of
infectivity and by PrPSc accumulation (18–22). Because low
levels of PrPSc are not associated with neuropathologic
changes, some discrepancy between vacuolation and PrPSc was
expected. In contrast to histology, IHC measures PrP
immunostaining after autoclaving tissue sections exposed to
formic acid. Because IHC measures PrP, we expected the
sensitivity of this procedure might be similar to the CDI,
but that proved not to be the case. Whether exposure of
formic acid-treated tissue sections to elevated temperature
destroys not only PrPC but also sPrPSc and only denatures
rPrPSc remains to be determined. Such a scenario could
account for the lower sensitivity of IHC compared with CDI
or bioassay (Tables 1 and 2).
Studies of the white matter in CJD brains were particularly
informative with respect to the sensitivity of the CDI,
where PrPSc levels were low but readily detectable, 10- to
100-fold above the threshold value (Fig. 4). Because animal
studies have shown that PrPSc and infectivity are
transported anterogradely from one brain region to another
along neuroanatomical pathways (23–25), we expected to find
PrPSc in white matter as demonstrated by the CDI but not
IHC. Axonal transport of PrPSc is also suggested by
diffusion-weighted MRI scans of CJD cases, which show
high-intensity signals in analogous neocortical regions of
the right and left cerebral hemispheres (26). This symmetry
of neuroradiological abnormalities is consistent with spread
of PrPSc to the contralateral cortex by means of callosal
commissural pathways.
Most immunoassays that detect HuPrPSc do so only after
subjecting the sample to limited proteolysis to form PrP
27–30, followed by denaturation. Because the CDI measures
the immunoreactivity before and after denaturation to an
epitope that is exposed in native PrPC but buried in PrPSc,
limited proteolysis to eliminate PrPC is unnecessary. Assays
based on limited proteolysis underestimate the level of
PrPSc because they digest sPrPSc, which represents 80–90% of
PrPSc in CJD and scrapie brains (Fig. 4 and Table 5).
Gerstmann–Sträussler–Scheinker, an inherited human prion
disease, is caused by the P102L mutation in the PRNP gene.
In mice expressing the Gerstmann–Sträussler–Scheinker mutant
PrP transgene, the CDI detected high levels of sPrPSc(P101L)
as well as low levels of rPrPSc(P101L) long before
neurodegeneration and clinical symptoms occurred (9).
sPrPSc(P101L) as well as low concentrations of rPrPSc(P101L)
previously escaped detection (27). Whether a similar
situation applies in other genetic forms of prion disease,
sCJD, or variant CJD remains to be determined. Because most
of the PrPSc in the brains of sCJD patients is
protease-sensitive (Fig. 4), it is likely that the lower
sensitivity of IHC is due to its inability to detect sPrPSc.
Presently, we have no information about the kinetics of
either sPrPSc or rPrPSc accumulation in human brain. Limited
information on the kinetics of PrPSc accumulation in
livestock comes from studies of cattle, sheep, and goats
inoculated orally, but most of the bioassays were performed
in non-Tg mice (28–30) in which prion titers were
underestimated by as much as a factor of 104 (10).
The studies reported here are likely to change profoundly
the approach to the diagnosis of prion disease in both
humans and livestock (31–33). The superior performance of
the CDI in diagnosing prion disease compared to routine
neuropathologic examination and IHC demands that the CDI be
used in future diagnostic evaluations of prion disease.
Prion disease can no longer be ruled out by routine
histology or IHC. Moreover, the use of IHC to confirm cases
of bovine spongiform encephalopathy after detection of
bovine PrPSc by the CDI (10) seems an untenable approach in
the future. Clearly, the CDI for HuPrPSc is as sensitive or
more sensitive than bioassays in Tg(MHu2M) mice (Fig. 1).
Our results suggest that using the CDI to test large numbers
of samples for human prions might alter the epidemiology of
prion diseases. At present, there is limited data on the
frequency of subclinical variant CJD infections in the U.K.
population (34). Because appendixes and tonsils were
evaluated only by IHC, many cases might have escaped
detection (Tables 1 and 2). Equally important may be the use
of CDI-like tests to diagnose other neurodegenerative
disorders, such as Alzheimer's disease, Parkinson's disease,
and the frontotemporal dementias. Whether IHC underestimates
the incidence of one or more of these common degenerative
diseases is unknown. Moreover, CDI-like tests may help
determine the frequency with which these disorders and the
prion diseases occurs concomitantly in a single patient (35,
36).
Acknowledgements
snip...END
http://www.pnas.org/
Volume 349:1812-1820 November 6, 2003 Number 19
Extraneural Pathologic Prion Protein in Sporadic Creutzfeldt-Jakob Disease
Background In patients with sporadic Creutzfeldt–Jakob disease, pathologic disease-associated prion protein (PrPSc) has been identified only in the central nervous system and olfactory-nerve tissue. Understanding the distribution of PrPSc in Creutzfeldt–Jakob disease is important for classification and diagnosis and perhaps even for prevention.
Methods We used a highly sensitive method of detection — involving the concentration of PrPSc by differential precipitation with sodium phosphotungstic acid, which increased the sensitivity of Western blot analysis by up to three orders of magnitude — to search for PrPSc in extraneural organs of 36 patients with sporadic Creutzfeldt–Jakob disease who died between 1996 and 2002.
Results PrPSc was present in the brain tissue of all patients. In addition, we found PrPSc in 10 of 28 spleen specimens and in 8 of 32 skeletal-muscle samples. Three patients had PrPSc in both spleen and muscle specimens. Patients with extraneural PrPSc had a significantly longer duration of disease and were more likely to have uncommon molecular variants of sporadic Creutzfeldt–Jakob disease than were patients without extraneural PrPSc.
Conclusions Using sensitive techniques, we identified extraneural deposition of PrPSc in spleen and muscle samples from approximately one third of patients who died with sporadic Creutzfeldt–Jakob disease. Extraneural PrPSc appears to correlate with a long duration of disease.
Source Information
From the Institute of Neuropathology and National Reference Center for Prion Diseases, University Hospital of Zurich, Zurich, Switzerland.
Dr. Glatzel and Mr. Abela contributed equally to the article.
Address reprint requests to Dr. Aguzzi at the Institute of Neuropathology, University Hospital of Zurich, Schmelzbergstr. 12, CH-8091 Zurich, Switzerland, or at [email protected] .
http://content.nejm.org/cgi/
Creutzfeldt-Jakob disease and inclusion body myositis: Abundant disease-associated prion protein in muscle
Gabor G. Kovacs, MD PhD 1 2, Elisabeth Lindeck-Pozza, MD 1, Leila Chimelli, MD, PhD 3, Abelardo Q. C. Araújo, MD, PhD 4, Alberto A. Gabbai, MD, PhD 5, Thomas Ströbel, PhD 1, Markus Glatzel, MD 6, Adriano Aguzzi, MD, PhD 6, Herbert Budka, MD 1 *
1Institute of Neurology, University of Vienna, and Austrian Reference Centre for Human Prion Diseases, Vienna, Austria
2National Institute of Psychiatry and Neurology, Budapest, Hungary
3Department of Pathology, School of Medicine, Federal University of Rio de Janeiro
4Department of Neurology, School of Medicine, Federal University of Rio de Janeiro
5Department of Neurology, School of Medicine, Federal University of Sao Paulo, Brazil
6Institute of Neuropathology, University Hospital of Zürich, Zürich, Switzerland
email: Herbert Budka ([email protected]. )
*Correspondence to Herbert Budka, Institute of Neurology, AKH 4J, Wühringer Gürtel 18-20, POB 48, A-1097 Vienna, Austria
Funded by:
European Union (EU) Project; Grant Number: TSELAB QLK2-CT-2002-81523
EU Concerted Action PRIONET; Grant Number: QLK2-2000-CT-00837
Abstract
Pathologicalprion protein (PrPSc) is the hallmark of prion diseases affecting primarily the central nervous system. Using immunohistochemistry, paraffin-embedded tissue blot, and Western blot, we demonstrated abundant PrPSc in the muscle of a patient with sporadic Creutzfeldt-Jakob disease and inclusion body myositis. Extraneural PrPC-PrPSc conversion in Creutzfeldt-Jakob disease appears to become prominent when PrPC is abundantly available as substrate, as in inclusion body myositis muscle.
--------------
Received: 16 June 2003; Revised: 11 September 2003; Accepted: 11 September 2003
Digital Object Identifier (DOI)
10.1002/ana.10813 About DOI
http://www3.interscience.wiley.com/
AS Professor Aguzzi kindly put it most recently ;
107
Vet Pathol 42:107 108 (2005)
Letters to the Editor
Editor:
Absence of evidence is not always evidence of absence. In the article Failure to detect prion protein (PrPres) by immunohistochemistry in striated muscle tissues of animals experimentally inoculated with agents of transmissible spongiform encephalopathy, recently published in Veterinary Pathology (41:78 81, 2004), PrPres was not detected in striated muscle of experimentally infected elk, cattle, sheep, and raccoons by immunohistochemistry (IHC). Negative IHC, however, does not exclude the presence of PrPSc. For example, PrPres was detected in skeletal muscle in 8 of 32 humans with the prion disease, sporadic Creutzfeldt-Jakob disease (CJD), using sodium phosphotungstic acid (NaPTA) precipitation and western blot.1 The NaPTA precipitation, described by Wadsworth et al.,3 concentrates the abnormal isoform of the prion, PrPres, from a large tissue homogenate volume before western blotting. This technique has increased the sensitivity of the western blot up to three orders of magnitude and could be included in assays to detect PrPres. Extremely conspicuous deposits of PrPres in muscle were detected by IHC in a recent case report of an individual with inclusion body myositis and CJD.2 Here, PrPres was detected in the muscle by immunoblotting, IHC, and paraf- fin-embedded tissue blot. We would therefore caution that, in addition to IHC, highly sensitive biochemical assays and bioassays of muscle are needed to assess the presence or absence of prions from muscle in experimental and natural TSE cases.
Christina Sigurdson, Markus Glatzel, and Adriano Aguzzi
Institute of Neuropathology
University Hospital of Zurich
Zurich, Switzerland
References
1 Glatzel M, Abela E, et al: Extraneural pathologic prion
protein in sporadic Creutzfeldt-Jakob disease. N Engl J
Med 349(19):1812 1820, 2003
2 Kovacs GG, Lindeck-Pozza E, et al: Creutzfeldt-Jakob
disease and inclusion body myositis: abundant diseaseassociated
prion protein in muscle. Ann Neurol 55(1):
121 125, 2004
3 Wadsworth JDF, Joiner S, et al: Tissue distribution of protease
resistant prion protein in variant CJD using a highly
sensitive immuno-blotting assay. Lancet 358:171 180,
2001...///
EMBO reports AOP Published online: 11 April 2003
Widespread PrPSc accumulation in muscles of hamsters orally infected with scrapie
http://www.emboreports.org/
Watson Meldrum et al Scrapie agent in muscle - Pattison I A (1990)
Veterinary record, 20 January 1990. p.68
http://www.bseinquiry.gov.uk/files/yb/1990/01/19009001.pdf
===============================
GREETINGS AGAIN APHIS ET AL,
FURTHERMORE, WE HAVE FAILED TO EVEN STOP THE SRMs FROM WHOLE
CUTS OF BONELESS BEEF IMPORTED FROM CANADA IN THE VERY ONSET OF
THE NEW BSE MRR (MINIMAL RISK REGION). THIS IS THE VERY REASON I HAVE
SAID TIME AND TIME AGAIN THAT BY THIS ADMINISTRATION ABANDONING THE
BSE GBR RISK ASSESSMENTS BECAUSE THEY DID NOT LIKE THE ASSESSMENT
OF BSE GBR III, AND ADHERING TO A NEW BSE MRR POLICY THAT WAS DESIGNED
NOT FOR HUMAN HEALTH, BUT ONLY FOR COMMODITIES AND FUTURES, WILL FURTHER
EXPOSE NEEDLESSLY MILLIONS AND MILLIONS OF HUMANS AND ANIMALS VIA THE FREE
TRADING OF ALL STRAINS OF TSE GLOBALLY. references as follow ;
Wisconsin Firm Recalls Beef Products
Recall Release CLASS II RECALL
FSIS-RC-032-2005 HEALTH RISK: LOW
Congressional and Public Affairs
(202) 720-9113
Steven Cohen
WASHINGTON, Aug. 19, 2005 - Green Bay Dressed Beef, a Green Bay, Wis., establishment, is voluntarily recalling approximately 1,856 pounds of beef products that may contain portions of the backbone from a cow just over 30 months old, the U.S. Department of Agriculture's Food Safety and Inspection Service announced today. The product was from a cow imported directly for slaughter from Canada.
Based on information provided by Canada, the products subject to this Class II recall are from a cow that is approximately one month older than the 30-month age limit. Both ante-mortem and post-mortem inspection were done on the cow in question. FSIS inspection program personnel determined the cow to be healthy and fit for human food. FSIS' designation of this recall as Class II is because it is a situation where there is a remote probability of adverse health consequences from the use of the product.
FSIS learned about this as a result of a Canadian audit of their health certificate that accompanied the imported cow. Prior to slaughter, the health certificate accompanying the cow was presented to the establishment, and it appeared complete and accurate. However, a subsequent audit of information related to the health certificate by Canadian officials found that it was not accurate. Action has been taken by Canadian Food Inspection Agency officials in response to findings from the audit.
The products subject to recall are:
Five boxes of 243 lb. vacuum pouched packages of "American Foods Group, NECKBONE UNTRIM'D, USDA CHOICE OR HIGHER" with the case code of 77333;
One box of 50 lb. vacuum pouched package of "American Foods Group, SHORTLOIN 2X2, USDA SELECT OR HIGHER" with the case code of 75231;
One box of 60 lb. vacuum pouched package of "American Foods Group, SHORTLOIN 2X2, USDA CHOICE OR HIGHER" with the case code of 75060;
Five boxes of 258 lb. vacuum pouched packages of "Dakota Supreme Beef, SHORTLOIN 0X11/4, USDA SELECT OR HIGHER" with the case code of 75442;
Sixteen boxes of 811 lb. vacuum pouched packages of "American Foods Group, BLADE BI N/O CHUCK, USDA CHOICE OR HIGHER" with the case code of 75955;
Nine boxes of 435 lb. vacuum pouched packages of "American Foods Group, BLADE BI N/O CHUCK, USDA SELECT OR HIGHER" with the case code of 75952.
Each box bears the establishment number "410" inside the USDA seal of inspection. The products were produced on August 4, and were distributed to wholesale distributors in Pennsylvania, Florida, Illinois, Maryland, Minnesota and Wisconsin.
Under the interim final rules FSIS implemented on January 12, 2004, certain specified risk materials must be removed from all cattle depending on the age of the animal. On this animal all specified risk materials for cattle 30 months and over were removed, with the exception of the vertebral column. At the time of slaughter, the animal was certified to be under 30 months of age and removal of the vertebral column was not required. A subsequent audit determined the animal was just over 30 months of age; therefore, the vertebral column is required to be removed. This is the reason for the recall of the selected products.
Consumers with questions about the recall may contact Sally VandeHei, Executive Assistant at 1-877-894-3927. National media with questions may contact Jim Mulhern at (202) 496-2468. Local media with questions may contact Susan Finco at (920) 965-7750 ext.158.
Consumers with other food safety questions can phone the toll-free USDA Meat and Poultry Hotline at 1-888-MPHotline (1-888-674-6854). The hotline is available in English and Spanish and can be reached from 10 a.m. to 4 p.m. (Eastern Time), Monday through Friday. Recorded food safety messages are available 24 hours a day.
Sample Product Labels: These are similar to, but not identical to, labels on the recalled product.
#
USDA Recall Classifications
Class I This is a health hazard situation where there is a reasonable probability that the use of the product will cause serious, adverse health consequences or death.
Class II This is a health hazard situation where there is a remote probability of adverse health consequences from the use of the product.
Class III This is a situation where the use of the product will not cause adverse health consequences.
http://www.fsis.usda.gov/News_&_Events/Recall_032_2005_Release/index.asp
:
Docket No. 03-080-1 -- USDA ISSUES PROPOSED RULE TO ALLOW LIVE ANIMAL IMPORTS FROM CANADA
[TSS SUBMISSION 11/03/2003 01:19 PM To: [email protected] ]
https://web01.aphis.usda.gov/BSEcom.nsf/0/b78ba677e2b0c12185256dd300649f9d?OpenDocument&AutoFramed
THE BSE MRR POLICY SHOULD BE ABOLISHED/REPEALED IMMEDIATELY AND THE BSE GBR RISK ASSESSMENTS AND POLICY SHOULD BE STRICTLY ENFORCED AND FURTHER ENHANCED TO INCLUDE CWD AND ALL TSEs...TSS
EFSA Scientific Report on the Assessment of the Geographical BSE-Risk (GBR) of the United States of America (USA)
Publication date: 20 August 2004
Adopted July 2004 (Question N° EFSA-Q-2003-083)
Report
Summary
Summary of the Scientific Report
The European Food Safety Authority and its Scientific Expert Working Group on the Assessment of the Geographical Bovine Spongiform Encephalopathy (BSE) Risk (GBR) were asked by the European Commission (EC) to provide an up-to-date scientific report on the GBR in the United States of America, i.e. the likelihood of the presence of one or more cattle being infected with BSE, pre-clinically as well as clinically, in USA. This scientific report addresses the GBR of USA as assessed in 2004 based on data covering the period 1980-2003.
The BSE agent was probably imported into USA and could have reached domestic cattle in the middle of the eighties. These cattle imported in the mid eighties could have been rendered in the late eighties and therefore led to an internal challenge in the early nineties. It is possible that imported meat and bone meal (MBM) into the USA reached domestic cattle and leads to an internal challenge in the early nineties.
A processing risk developed in the late 80s/early 90s when cattle imports from BSE risk countries were slaughtered or died and were processed (partly) into feed, together with some imports of MBM. This risk continued to exist, and grew significantly in the mid 90's when domestic cattle, infected by imported MBM, reached processing. Given the low stability of the system, the risk increased over the years with continued imports of cattle and MBM from BSE risk countries.
EFSA concludes that the current GBR level of USA is III, i.e. it is likely but not confirmed that domestic cattle are (clinically or pre-clinically) infected with the BSE-agent. As long as there are no significant changes in rendering or feeding, the stability remains extremely/very unstable. Thus, the probability of cattle to be (pre-clinically or clinically) infected with the BSE-agent persistently increases.
http://www.efsa.eu.int/science/efsa_scientific_reports/gbr_assessments/573_en.html
From: Terry S. Singeltary Sr. [[email protected]]
Sent: Tuesday, July 29, 2003 1:03 PM
To: [email protected]
Cc: [email protected]; [email protected].; BSE-L
Subject: Docket No. 2003N-0312 Animal Feed Safety System [TSS SUBMISSION
TO DOCKET 2003N-0312]
Greetings FDA,
snip...
PLUS, if the USA continues to flagrantly ignore the _documented_ science to date about the known TSEs in the USA (let alone the undocumented TSEs in cattle), it is my opinion, every other Country that is dealing with BSE/TSE should boycott the USA and demand that the SSC reclassify the USA BSE GBR II risk assessment to BSE/TSE GBR III 'IMMEDIATELY'. for the SSC to _flounder_ any longer on this issue, should also be regarded with great suspicion as well. NOT to leave out the OIE and it's terribly flawed system of disease surveillance. the OIE should make a move on CWD in the USA, and make a risk assessment on this as a threat to human health. the OIE should also change the mathematical formula for testing of disease. this (in my opinion and others) is terribly flawed as well. to think that a sample survey of 400 or so cattle in a population of 100 million, to think this will find anything, especially after seeing how many TSE tests it took Italy and other Countries to find 1 case of BSE (1 million rapid TSE test in less than 2 years, to find 102 BSE cases), should be proof enough to make drastic changes of this system. the OIE criteria for BSE Country classification and it's interpretation is very problematic. a text that is suppose to give guidelines, but is not understandable, cannot be considered satisfactory. the OIE told me 2 years ago that they were concerned with CWD, but said any changes might take years. well, two years have come and gone, and no change in relations with CWD as a human health risk. if we wait for politics and science to finally make this connection, we very well may die before any decisions
or changes are made. this is not acceptable. we must take the politics and the industry out of any final decisions of the Scientific community. this has been the problem from day one with this environmental man made death sentence. some of you may think i am exaggerating, but you only have to see it once, you only have to watch a loved one die from this one time, and you will never forget, OR forgive...yes, i am still very angry... but the transmission studies DO NOT lie, only the politicians and the industry do... and they are still lying to this day...TSS
http://www.fda.gov/ohrms/dockets/dockets/03n0312/03N-0312_emc-000001.txt
GREETINGS AGAIN APHIS ET AL,
Moving on to the theory that BSE agent is not in blood. THIS is what they use to think with nvCJD. However
the nvCJD agent has now been detected and transmitted the TSE agent by blood. nvCJD is the BSE agent
that has transmitted to humans. nvCJD is human BSE. so if nvCJD transmits by blood, why not BSE? with
the limited testing to date, the limited sensitivity of the detection of the BSE/TSE agent blood to date, i would
not be so sure that the BSE/TSE agent does not transmit by blood. just recent Ag. Comm. Johanns stated
that they would not address the blood issue being fed to cattle. a foolish and careless mistake. but typical.
we now have detected new atypical strains of the BSE/TSE agent in cattle in many countries i.e. Japan, France,
Belgium, Germany, and Italy. In the Italian study of BASE, a new? TSE in cattle they have discovered that is
not like the nvCJD, but very similar to sporadic CJD. They have detected 2 such cattle at printing of this study
March 2, 2004. Identification of a second bovine amyloidotic spongiform encephalopathy: Molecular similarities
with sporadic Creutzfeldt-Jakob disease PNAS. MY point is that with these new atypical TSEs showing up in
cattle, sheep and goats, we don't know if blood and other tissues transmits the disease. THE SRM list may
and should be reevaluated. WE know BSE is in the USA, but we also know that in the studies of Mission Texas,
where USA suffolk scrapie sheep were inoculated into USA cattle, the PRION agent that was produced did not
look like the UK BSE strain. so why would all CJD cases in the USA look like the UK human BSE i.e. nv/v CJD?
WHY wait and expose millions and millions needlessly as in the past with previous TSE blunders, why wait to
act. why not act first with what we know, which is very concerning, then as science evolves, go from there.
references as follow ;
UK Strategy for Research and
Development on Human and Animal
Health Aspects of Transmissible
Spongiform Encephalopathies
2005-2008
snip...
3.2 Tackling the spread of infection
3.2.1 The species barrier and the carrier state
3.2.1.1 The possibility of 'carrier' states in animals and humans, and our present inability
to identify them, pose a potential threat to public and animal health. The susceptibility of
humans to BSE infection, and the ability of the disease to remain clinically silent for
many years, is of major concern to DH. Although the death of a UK blood donor from
vCJD in 1999 three years after making the donation and the subsequent death from vCJD
of the recipient in 2003 have not been causally linked, transmission of infection through
blood transfusion is the most likely explanation327. The case heightens concerns that
'carriers' could be transmitting the disease through blood, tissue and organ donation or
by contaminating surgical instruments when undergoing surgery. DH will continue to
support research to detect infectious prions in human tissue, to investigate the
decontamination of surgical instruments and to develop measures to protect blood
supplies.
3.2.1.2 Animal models of some TSEs have detected infectivity in blood. Experiments,
which have involved transfusing large volumes of blood from infected sheep to healthy
recipient sheep, have demonstrated that infectivity can be transmitted by blood
transfusion. A central part of DH policy in this area has been the leucodepletion of blood
donations and the efficacy of this technology can now be tested in sheep.
snip...
4.5.4 In 1987, epidemiological studies of BSE cases identified meat and bone meal as the
probable means by which the disease was being spread. In an attempt to prevent
further infections a ban on incorporating ruminant protein in ruminant feed was
introduced in July 1988. Due to the long incubation period associated with this disease
the efficacy of this control measure was not immediately apparent. As time passed it
became clear from the number of cases born after the ban that it was not wholly
effective.
4.5.5 Epidemiological analysis of these cases showed that a high proportion of them
occurred in areas where the pig population was high. This observation, coupled with
research data that showed that only a very small dose of the infective material was
needed to cause disease in cattle, led to the conclusion that cross-contamination of feed
was occurring.
4.5.6 Since 1988, increasingly stringent feed controls have been put in place. Key
amongst these have been:
• the ban on the use of specified bovine offal in all animal feed (September 1990);
• the ban on feeding any farmed animal, including horses and fish, with mammalian
meat and bone meal. (This began in March 1996 but following this ban there was
a recall scheme and the date from which the ban was considered to be fully
effective is regarded as being 1 August 1996);
62
v6.1
• EU-wide controls on feed which extended the ban to include all processed animal
protein, including that derived from birds and fish (implemented in the UK from 1
August 2001).
4.5.7 As illustrated in fig. [ ] these later measures have reduced the spread of BSE.
However, they have not been one hundred per cent effective. As at 31st December 2003
there had been 81 cases of BSE in animals born since 1 August 1996 in the UK.
full text 91 pages;
http://www.mrc.ac.uk/pdf-about-tse_uk_strategy_june2005.pdf
GREETINGS AGAIN APHIS ET AL,
JUST what about those old studies at Mission Texas and the atypical TSE in cattle?
would it not be prudent for human health purpose, the question that, with all the atypical
TSEs showing up in animal and man in different countries, the fact that most all of these
TSEs transmit as freely or not as freely as BSE (depending whom you have witnessed die from
this agent either directly or indirectly via a multitude of potential routes and sources) to primates.
would it not be prudent to ask yourself if some if not all of these sporadic CJDs might be a
by-product of these TSEs either directly or indirectly via a multitude of proven routes and sources
in animal studies? it is unethical for human transmission studies considering the fact that the agent
is 100% fatal, slow, but fatal. there has been no sound science in any of the recent decisions in the
USA in regards to BSE/TSE human or animal, all one has to do is look at TEXAS, the mad cow that
got away, the stumbling and staggering one that NO TSE TEST AT ALL was done, ordered rendered,
and then the infamous positive, positve, secret positive, inconclusive (NO WB), negative, 8 month delay,
then the 'Fong Effect' took place, THEN FINALLY CONFIRMED SOME 8 MONTHS LATER IN WEYBRIDGE.
Finally recently, another inconclusive that took place that sat untested on some shelf for about 4 months while
the Texas mad cow blunder was going on. the tissues of this cow this time were preserved in preservative as
to render any WB for further confirmation, what i called the 'FONG SYNDROME' or the 'end around' the WB
ordered previously by the Honorable Phyllis Fong of the OIG. Politics at its finest, to hell with human health.
WE find now that 9,200 USA POTENTIAL MAD COWS IN JUNE 2004 ENHANCED COVER-UP
SURVEILLANCE PROGRAM WENT UNTESTED WITH NO RAPID TEST OR WB, ONLY IHC, the test
that fails the most, that is very unreliable as noted above in my submission by Prusiner et al and other scientist.
NOW, back to Mission, Texas ;
>> Differences in tissue distribution could require new regulations
>> regarding specific risk material (SRM) removal.
snip...end
full text 33 PAGES ;
http://www.bseinquiry.gov.uk/files/mb/m11b/tab01.pdf
http://www.bseinquiry.gov.uk/files/yb/1988/10/00001001.pdf
It was, however, performed in the USA in 1979, when it was shown that cattle
inoculated with the scrapie agent endemic in the flock of Suffolk sheep at
the United States Department of Agriculture in Mission, Texas, developed a
TSE quite unlike BSE. 32 The findings of the initial transmission, though
not of the clinical or neurohistological examination, were communicated in
October 1988 to Dr Watson, Director of the CVL, following a visit by Dr
Wrathall, one of the project leaders in the Pathology Department of the CVL,
to the United States Department of Agriculture. 33 The results were not
published at this point, since the attempted transmission to mice from the
experimental cow brain had been inconclusive. The results of the clinical
and histological differences between scrapie-affected sheep and cattle were
published in 1995. Similar studies in which cattle were inoculated
intracerebrally with scrapie inocula derived from a number of
scrapie-affected sheep of different breeds and from different States, were
carried out at the US National Animal Disease Centre. 34 The results,
published in 1994, showed that this source of scrapie agent, though
pathogenic for cattle, did not produce the same clinical signs of brain
lesions characteristic of BSE.
http://www.bseinquiry.gov.uk/
1: J Infect Dis. 1994 Apr;169(4):814-20.
Intracerebral transmission of scrapie to cattle.
Cutlip RC, Miller JM, Race RE, Jenny AL, Katz JB, Lehmkuhl HD, DeBey BM,
Robinson MM.
USDA, Agriculture Research Service, National Animal Disease Center, Ames, IA
50010.
To determine if sheep scrapie agent(s) in the United States would induce a
disease in cattle resembling bovine spongiform encephalopathy, 18 newborn
calves were inoculated intracerebrally with a pooled suspension of brain
from 9 sheep with scrapie. Half of the calves were euthanatized 1 year after
inoculation. All calves kept longer than 1 year became severely lethargic
and demonstrated clinical signs of motor neuron dysfunction that were
manifest as progressive stiffness, posterior paresis, general weakness, and
permanent recumbency. The incubation period was 14-18 months, and the
clinical course was 1-5 months. The brain from each calf was examined for
lesions and for protease-resistant prion protein. Lesions were subtle, but a
disease-specific isoform of the prion protein was present in the brain of
all calves. Neither signs nor lesions were characteristic of those for
bovine spongiform encephalopathy.
MeSH Terms:
Animals
Brain/microbiology*
Brain/pathology
Cattle
Cattle Diseases/etiology*
Cattle Diseases/pathology
Encephalopathy, Bovine Spongiform/etiology*
Encephalopathy, Bovine Spongiform/pathology
Immunoblotting/veterinary
Immunohistochemistry
Male
Motor Neurons/physiology
Prions/analysis
Scrapie/pathology
Scrapie/transmission*
Sheep
Sleep Stages
Time Factors
Substances:
Prions
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8133096&dopt=Citation
Intracerebral transmission of scrapie to cattle FULL TEXT PDF;
SNIP...
Discussion
WE conclude that American sources of sheep scrapie are transmissible to
cattle by direct intracerebral inoculation but the disease induced is NOT
identical to BSE as seen in the United Kingdom. While there were
similarities in clinical signs between this experimental disease and BSE,
there was no evidence of a
From: Terry S. Singeltary Sr.
To: [email protected]. ; [email protected].
Sent: Thursday, August 25, 2005 9:16 AM
Subject: Importation of Whole Cuts of Boneless Beef from Japan [Docket No. 05-004-1] RIN 0579-AB93 TSS SUBMISSION
Greetings Dr. Colgrove and Miss Johnson,
Thank you for taking this submission via email. i have had trouble submitting via the comment page due to the length of my submission. I was not sure that my file attachment that i submitted via the ;
EDOCKET: Go to http://www.epa.gov/feddocket
I submitted yesterday, just did not know if the file reached anyone. so to make sure, I am sending to you to submit for me.
many thanks,
Terry
From: TSS ()
Subject: Importation of Whole Cuts of Boneless Beef from Japan [Docket No. 05-004-1] RIN 0579-AB93 TSS SUBMISSION
Date: August 24, 2005 at 2:47 pm PST
August 24, 2005
Importation of Whole Cuts of Boneless Beef from Japan [Docket No. 05-004-1] RIN 0579-AB93 TSS SUBMISSION
Greetings APHIS ET AL,
My name is Terry S. Singeltary Sr.
I would kindly like to comment on [Docket No. 05-004-1] RIN 0579-AB93 ;
PROPOSED RULES
Exportation and importation of animals and animal products:
Whole cuts of boneless beef from-
Japan,
48494-48500 [05-16422]
[Federal Register: August 18, 2005 (Volume 70, Number 159)]
[Proposed Rules]
[Page 48494-48500]
From the Federal Register Online via GPO Access [wais.access.gpo.gov]
[DOCID:fr18au05-7]
========================================================================
Proposed Rules
Federal Register
________________________________________________________________________
This section of the FEDERAL REGISTER contains notices to the public of
the proposed issuance of rules and regulations. The purpose of these
notices is to give interested persons an opportunity to participate in
the rule making prior to the adoption of the final rules.
========================================================================
[[Page 48494]]
DEPARTMENT OF AGRICULTURE
Animal and Plant Health Inspection Service
9 CFR Part 94
[Docket No. 05-004-1]
RIN 0579-AB93
Importation of Whole Cuts of Boneless Beef from Japan
AGENCY: Animal and Plant Health Inspection Service, USDA.
ACTION: Proposed rule.
-----------------------------------------------------------------------
SUMMARY: We are proposing to amend the regulations governing the
importation of meat and other edible animal products by allowing, under
certain conditions, the importation of whole cuts of boneless beef from
Japan. We are proposing this action in response to a request from the
Government of Japan and after conducting an analysis of the risk that
indicates that such beef can be safely imported from Japan under the
conditions described in this proposal.
DATES: We will consider all comments that we receive on or before
September 19, 2005.
ADDRESSES: You may submit comments by any of the following methods:
EDOCKET: Go to http://www.epa.gov/feddocket to submit or
snip...
BSE infectivity has never been demonstrated in the muscle tissue of
cattle experimentally or naturally infected with BSE at any stage of
the disease. Studies performed using TSEs other than BSE in non-bovine
animals have detected prions in muscle tissue. However, the
international scientific community largely considers that these studies
cannot be directly extrapolated to BSE in cattle because of the
significant interactions between the host species and the prion strain
involved.
Pathogenesis studies of naturally and experimentally infected
cattle have not detected BSE infectivity in blood. However,
transmission of BSE was demonstrated in sheep that received a
transfusion of a large volume of blood drawn from other sheep that were
experimentally infected with the BSE agent. The United Kingdom's
Department for Environment, Food and Rural Affairs' Spongiform
Encephalopathy Advisory Committee (SEAC) and the European Commission's
Scientific Steering Committee (SSC), which are scientific advisory
committees, evaluated the implication of this finding in relation to
food safety.\5\ The SEAC concluded that the finding did not represent
grounds for recommending any changes to the current control measures
for BSE. The SSC determined that the research results do not support
the hypothesis that bovine blood or muscle meat constitute a risk to
human health.\6\
snip...
BSE Risk Factors for Whole Cuts of Boneless Beef
The most significant risk management strategy for ensuring the
safety of whole cuts of boneless beef is the prevention of cross-
contamination of the beef with SRMs during stunning and slaughter of
the animal. Control measures that prevent contamination of such beef
involve the establishment of procedures for the removal of SRMs,
prohibitions on air-injection stunning and pithing, and splitting of
carcasses. These potential pathways for contamination and the control
measures that prevent contamination are described in detail in the risk
analysis for this rulemaking.
SRM Removal. Research has demonstrated that SRMs from infected
cattle may contain BSE infectivity. Because infectivity has not been
demonstrated in muscle tissue, the most important mitigation measure
for whole cuts of boneless beef is the careful removal and segregation
of SRMs. Removal of SRMs in a manner that avoids contamination of the
beef with SRMs minimizes the risk of exposure to materials that have
been demonstrated to contain the BSE agent in cattle.
snip...
Variant Creutzfeldt-Jakob disease (vCJD), a chronic and fatal
neurodegenerative disease of humans, has been linked since 1996 through
epidemiological, neuropathological, and experimental data to exposure
to the BSE agent, most likely through consumption of cattle products
contaminated with the agent before BSE control measures were in place.
To date, approximately 170 probable and confirmed cases of vCJD have
been identified worldwide. The majority of these cases have either been
identified in the United Kingdom or were linked to exposure that
occurred in the United Kingdom, and all cases have been linked to
exposure in countries with native cases of BSE. Some studies estimate
that more than 1 million cattle may have been infected with BSE
throughout the epidemic in the United Kingdom. This number of infected
cattle could have introduced a significant amount of infectivity into
the human food supply. Yet, the low number of cases of vCJD identified
to date indicates that there is a substantial species barrier that
protects humans from widespread illness due to exposure to the BSE
agent.
snip...
International Guidelines on BSE
International guidelines for trade in animal and animal products
are developed by the World Organization for Animal Health (formerly
known as the Office International des Epizooties (OIE)), which is
recognized by the World Trade Organization (WTO) as the international
organization responsible for the development of standards, guidelines,
and recommendations with respect to animal health and zoonoses
(diseases that are transmissible from animals to humans). The OIE
guidelines for trade in terrestrial animals (mammals, birds, and bees)
are detailed in the Terrestrial Animal Health Code (available on the
internet at http://www.oie.int). The guidelines on BSE are contained in
Chapter 2.3.13 of the Code and supplemented by Appendix 3.8.4 of the
Code.
snip...end
http://a257.g.akamaitech.net/7/257/2422/01jan20051800/edocket.access.gpo.gov/2005/05-16422.htm http://a257.g.akamaitech.net/7/257/2422/01jan20051800/edocket.access.gpo.gov/2005/pdf/05-16422.pdf
Greetings again APHIS ET AL,
THIS is not correct. IN fact, there are several factors i would like to kindly address.
Muscle tissue has recently been detected with PrPSc
in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve) of the 11th BSE
cow in Japan (Yoshifumi Iwamaru et al). also recently, Aguzzi et al Letter to the Editor
Vet Pathol 42:107-108 (2005), Prusiner et al CDI test is another example of detection
of the TSE agent in muscle in sCJD, Herbert Budka et al CJD and inclusion body myositis:
Abundant Disease-Associated Prion Protein in Muscle, and older studies from Watson
Meldrum et al Scrapie agent in muscle - Pattison I A (1990), references as follow ;
PrPSc distribution of a natural case of bovine
spongiform encephalopathy
Yoshifumi Iwamaru, Yuka Okubo, Tamako Ikeda, Hiroko Hayashi, Mori-
kazu Imamura, Takashi Yokoyama and Morikazu Shinagawa
Priori Disease Research Center, National Institute of Animal Health, 3-1-5
Kannondai, Tsukuba 305-0856 Japan [email protected]
Abstract
Bovine spongiform encephalopathy (BSE) is a disease of cattle that causes
progressive neurodegeneration of the central nervous system. Infectivity
of BSE agent is accompanied with an abnormal isoform of prion protein
(PrPSc).
The specified risk materials (SRM) are tissues potentially carrying BSE
infectivity. The following tissues are designated as SRM in Japan: the
skull including the brain and eyes but excluding the glossa and the masse-
ter muscle, the vertebral column excluding the vertebrae of the tail, spinal
cord, distal illeum. For a risk management step, the use of SRM in both
animal feed or human food has been prohibited. However, detailed
PrPSc distribution remains obscure in BSE cattle and it has caused con-
troversies about definitions of SRM. Therefore we have examined PrPSc
distribution in a BSE cattle by Western blotting to reassess definitions of
SRM.
The 11th BSE case in Japan was detected in fallen stock surveillance.
The carcass was stocked in the refrigerator. For the detection of PrPSc,
200 mg of tissue samples were homogenized. Following collagenase
treatment, samples were digested with proteinase K. After digestion,
PrPSc was precipitated by sodium phosphotungstate (PTA). The pellets
were subjected to Western blotting using the standard procedure.
Anti-prion protein monoclonal antibody (mAb) T2 conjugated horseradish
peroxidase was used for the detection of PrPSc.
PrPSc was detected in brain, spinal cord, dorsal root ganglia, trigeminal
ganglia, sublingual ganglion, retina. In addition, PrPSc was also detected
in the peripheral nerves (sciatic nerve, tibial nerve, vagus nerve).
Our results suggest that the currently accepted definitions of SRM in
BSE cattle may need to be reexamined. ...
179
T. Kitamoto (Ed.)
PRIONS
Food and Drug Safety
================
ALSO from the International Symposium of Prion Diseases held in Sendai, October 31, to
November 2, 2004;
Bovine spongiform encephalopathy (BSE) in Japan
snip...
"Furthermore, current studies into transmission of cases of BSE that are
atypical or that develop in young cattle are expected to amplify the BSE
prion"
NO. Date conf. Farm Birth place and Date Age at diagnosis
8. 2003.10.6. Fukushima Tochigi 2001.10.13. 23
9. 2003.11.4. Hiroshima Hyogo 2002.1.13. 21
Test results
# 8b, 9c cows Elisa Positive, WB Positive, IHC negative, histopathology
negative
b = atypical BSE case
c = case of BSE in a young animal
b,c, No PrPSc on IHC, and no spongiform change on histology
International Symposium of Prion Diseases held in Sendai, October 31, to
November 2, 2004.
The hardback book title is 'PRIONS' Food and Drug Safety
T. Kitamoto (Ed.)
Tetsuyuki Kitamoto
Professor and Chairman
Department of Prion Research
Tohoku University School of Medicine
2-1 SeiryoAoba-ku, Sendai 980-8575, JAPAN
TEL +81-22-717-8147 FAX +81-22-717-8148
e-mail; [email protected]
Symposium Secretariat
Kyomi Sasaki
TEL +81-22-717-8233 FAX +81-22-717-7656
e-mail: [email protected]
================================
107
Vet Pathol 42:107–108 (2005)
Letters to the Editor
Editor:
Absence of evidence is not always evidence of absence.
In the article ''Failure to detect prion protein (PrPres) by
immunohistochemistry in striated muscle tissues of animals
experimentally inoculated with agents of transmissible spongiform
encephalopathy,'' recently published in Veterinary
Pathology (41:78–81, 2004), PrPres was not detected in striated
muscle of experimentally infected elk, cattle, sheep, and
raccoons by immunohistochemistry (IHC). Negative IHC,
however, does not exclude the presence of PrPSc. For example,
PrPres was detected in skeletal muscle in 8 of 32
humans with the prion disease, sporadic Creutzfeldt-Jakob
disease (CJD), using sodium phosphotungstic acid (NaPTA)
precipitation and western blot.1 The NaPTA precipitation,
described by Wadsworth et al.,3 concentrates the abnormal
isoform of the prion, PrPres, from a large tissue homogenate
volume before western blotting. This technique has increased
the sensitivity of the western blot up to three orders
of magnitude and could be included in assays to detect
PrPres. Extremely conspicuous deposits of PrPres in muscle
were detected by IHC in a recent case report of an individual
with inclusion body myositis and CJD.2 Here, PrPres was
detected in the muscle by immunoblotting, IHC, and paraf-
fin-embedded tissue blot. We would therefore caution that,
in addition to IHC, highly sensitive biochemical assays and
bioassays of muscle are needed to assess the presence or
absence of prions from muscle in experimental and natural
TSE cases.
Christina Sigurdson, Markus Glatzel, and Adriano Aguzzi
Institute of Neuropathology
University Hospital of Zurich
Zurich, Switzerland
References
1 Glatzel M, Abela E, et al: Extraneural pathologic prion
protein in sporadic Creutzfeldt-Jakob disease. N Engl J
Med 349(19):1812–1820, 2003
2 Kovacs GG, Lindeck-Pozza E, et al: Creutzfeldt-Jakob
disease and inclusion body myositis: abundant diseaseassociated
prion protein in muscle. Ann Neurol 55(1):
121–125, 2004
3 Wadsworth JDF, Joiner S, et al: Tissue distribution of protease
resistant prion protein in variant CJD using a highly
sensitive immuno-blotting assay. Lancet 358:171–180,
2001
===================================
Corinna Kaarlela, News Director
Source: Jennifer O'Brien
[email protected]
415-476-2557
14 February 2005
Diagnosis of prions in patients should utilize novel strategy, team says
A technique for detecting prions in tissue, developed in recent years by
UCSF scientists, is significantly more sensitive than the diagnostic
procedures currently used to detect the lethal particles in samples of
brain tissue from patients, according to a study performed by a UCSF team.
The finding indicates that the diagnostic technique, known as the
conformation-dependent immunoassay (CDI), should be established as the
standard approach for brain biopsies of patients suspected of having the
disease, they say. The team is exploring whether the CDI might be
adapted to detect prions in blood and muscle.
The finding suggests that reliance on the current methods for detecting
prions in human brain tissue -- microscopic examination of tissue for
the telltale vacuoles that form in brain cells and immunohistochemistry
(IHC), which involves detecting prions in brain sections using prion
protein-specific antibodies -- may have led to an under diagnosis of the
disease in patients in recent years, they say. (A definitive diagnosis
of the disease in humans is made only on autopsy, when a
neuropathologist can analyze multiple brain regions for vacuoles and
evidence of prions by IHC, and it is estimated that only 50 percent of
human cases are autopsied, in part because many pathologists do not want
to risk infection during the autopsy.)
In the study, the team compared the ability of the CDI and the two
traditional diagnostic techniques to detect prions in various brain
samples from 28 patients diagnosed on autopsy as having one of several
human forms of the disease -- sporadic, familial or iatrogenic
Creutzfeldt-Jakob disease (CJD). While the CDI detected the biochemical
signal for prions in 100 percent of the samples studied, the traditional
tests failed to detect the prion in a high proportion of cases. For
example, in an experiment that focused on 18 brain regions from eight
patients with sporadic CJD, the CDI detected prions in 100 percent of
the samples, while IHC detected them in 22 percent and routine tissue
examination in 17 percent.
"In about 80 percent of the different brain regions examined, prions
were not consistently detected by either IHC or routine histology that
measure vacuolation. In contrast, the CDI was always positive in all
regions of the brain," says the lead author of the study, Jiri Safar,
MD, associate adjunct professor of neurology and a member of the UCSF
Institute for Neurodegenerative Diseases, which is directed by senior
author Stanley B. Prusiner, MD, UCSF professor of neurology and
biochemistry.
"These findings indicate that histology and immunohistochemistry should
no longer be used to rule out prion disease in single-site biopsy
samples," says Safar. "The superior performance of the CDI in diagnosing
prion disease suggests that the CDI be used in future diagnostic
evaluations of prion disease, particularly for single-site brain
biopsies during life"
"If the traditional techniques are used at autopsy, they must be applied
to many cortical and subcortical samples," says co-author Stephen J.
DeArmond, MD, PhD, UCSF professor of neuropathology.
Moreover, while the study examined the efficacy of the CDI in comparison
to the two techniques routinely used by neuropathologists to detect
prions in human brain tissue, previous studies at UCSF indicate that the
CDI is also significantly more sensitive than Western blot analysis, the
technology used with IHC to detect prions in brain tissue from cattle
suspected of having bovine spongiform encephalopathy (BSE). That IHC and
Western blot analysis are relatively insensitive methods, the
researchers say, supports their ongoing assertion that the CDI should
also be used to evaluate the brain tissue of cattle.
"The studies reported here are likely to change profoundly the approach
to the diagnosis of prion disease in both humans and livestock," says Safar.
More broadly, the scientists say, the high sensitivity of the CDI
suggests that CDI-like tests could also prove useful for diagnosing
other neurodegenerative diseases, such as Alzheimer's disease,
Parkinsons's disease and fronto-temporal dementias, all of which, like
prion diseases, involve various forms of protein misprocessing. These
diseases currently are diagnosed by neuropathological analysis and
immunohistochemistry.
"Whether immunohistochemistry underestimates the incidence of one or
more of these common neurodegenerative diseases is unknown, but the CDI
could shed light on these diseases," says co-author Bruce Miller, MD,
UCSF A.W. and Mary Margaret Clausen Distinguished Professor of Neurology
and director of the UCSF Memory and Aging Center.
The finding will be printed on-line and in print on March 1, 2005 in
Proceedings of the National Academy of Sciences.
The study brings into high relief the different detection strategies of
immunohistochemistry and the CDI, both of which involve revealing the
presence of prions, known as PrPsc, by applying antibodies to brain tissue.
Standard immunohistochemistry, developed in the DeArmond lab 20 years
ago, involves using an enzyme known as a protease, or a combination of
harsh acid and high temperature treatment, to destroy normal prion
protein (PrPC), which is ubiquitous in brain tissue. Once this occurs,
scientists apply fluorescently lit antibodies that react with residues
of the relatively resistant abnormal prion protein (PrPSc), thereby
highlighting it.
The limitation of this technique is that scientists have since learned
that there is a large part of the abnormal prion protein that is
protease sensitive, and that portion escapes detection by the standard
technique. Thus, this traditional method underestimates the level of
PrPSc in tissue.
The CDI addresses this limitation by revealing the region of PrPSc that
is exposed in the normal PrPC but is buried in infectious PrPSc, using
high affinity, newly generated antibodies that identify PrPSc through
the distinct shape of the molecule, independent of proteolytic
treatments. This makes it possible to detect potentially large
concentrations of protease sensitive PrPSc molecules.
Detractors would say that it is not necessary to detect the minute level
of infectious agent that the CDI is capable of revealing, as it would be
unlikely to be lethal, says Safar. But Prusiner and his colleagues
maintain that any risk is too great when it comes to having prions in
the food supply. In addition, because even low levels of prions are
extremely resistant to inactivation, they may contaminate the
environment for many years.
Prusiner won the 1997 Nobel Prize in Physiology or Medicine for
discovering that a class of neurodegenerative diseases known as
spongiform encephalopathies was caused by prions. Prion diseases develop
in humans, cattle, sheep, deer, elk and mink.
The CDI was developed by members of the Prusiner lab. The CDI
methodology has been licensed to InPro Biotechnology, Inc.
Prusiner, Safar, DeArmond and other members of the Institute for
Neurodegenerative Diseases are scientific advisors to, or own stock in,
InPro.
Other co-authors of the study were Michael D. Geschwind, Camille
Deering, Svetlana Didorenko, Mamta Sattavat, Henry Sanchesz, Ana Serban,
Kurt Giles, of UCSF, and Martin Vey, of Behring, Marburg, Germany, and
Henry Baron, of Behring, Paris.
The study was funded by the National Institutes of Health, the John
Douglas French Foundation for Alzheimer's research, the McBean
Foundation, the State of California, Alzheimer's Disease Research Center
of California and the RR00079 General Clinical Research Center.
The UCSF Institute for Neurodegenerative Diseases:
http://ind.medschool.ucsf.edu/.
FURTHER COMPARISON OF THE CDI TO THE STANDARD DIAGNOSTIC PROCEDURES,
PROVIDED BY STEPHEN J. DEARMOND, MD, PHD, UCSF PROFESSORS OF NEUROPATHOLOGY:
Explanation as to why the CDI is more sensitive than Western blot
analysis: Studies at UCSF during development of the CDI showed that CDI
could detect prions in brain homogenates at levels that fail to produce
disease in animals (bioassay for prions). Therefore, the CDI is more
sensitive than the bioassay method, which was considered to be the most
sensitive technique for detecting prions. In contrast, Western blot
analysis for prions is significantly less sensitive than the bioassay
and is, therefore, significantly less sensitive than the CDI. Currently,
the USDA uses a combination of Western blot analysis of brainstem
homogenates and immunohistochemistry of the medulla to test cattle
suspected of having bovine spongiform encephalopathy ("mad cow
disease"). The relative insensitivity of IHC and Western blot analysis,
says DeArmond, supports the UCSF scientists' ongoing assertion that the
CDI should also be used to evaluate the brain tissue of cattle.
DeArmond cites additional evidence about Western blot analysis from a
World Health Organization (WHO) study group, which compared the CDI
method with Western blots for detection of prions in sporadic and
variant CJD brains. Based on the smallest amount of prions that could
detected by the two techniques, they found that the CDI was from 1000-
to 100,000-fold more sensitive than Western blot analysis performed in
six different research laboratories (Minor et al. Standards for the
assay of Creutzfeldt-Jakob disease specimens. J. Gen. Virol. 85:
1777-1784, 2004).
Explanation as to why IHC for prions is less sensitive than the CDI: IHC
is routinely performed on formalin-fixed, paraffin-embedded samples of
brain. Formalin fixation markedly decreases the ability of antibodies to
bind to proteins in general, which greatly weakens the IHC signal for
prions (PrPSc). In contrast, homogenates for the CDI are not treated
with reagents that decrease prion antigenicity. Moreover, to concentrate
the PrPSc for measurement by the CDI, the homogenates are exposed to
phosphotungstic acid, which selectively precipitates both
protease-sensitive and protease-resistant PrPSc that comprise prions,
but not the normal prion protein conformer found in uninfected animals,
PrPC. This step results in a higher concentration of PrPSc for detection
by the CDI. Because the PrPSc was not exposed to proteases, the CDI
measures all forms of abnormally folded PrPSc molecules.
Protease-sensitive PrPSc can account for 50 percent of the total PrPSc.
For Western analysis, homogenates of brain are treated with protease to
eliminate PrPC; however, this step also eliminates protease-sensitive
PrPSc leaving only protease-resistant PrPSc for Western blot detection
and decreasing the PrPSc signal at least in half.
###
http://pub.ucsf.edu/newsservices/releases/200502147/
PNAS | March 1, 2005 | vol. 102 | no. 9 | 3501-3506
NEUROSCIENCE
Diagnosis of human prion disease
Jiri G. Safar *, , Michael D. Geschwind , , Camille Deering
*, Svetlana Didorenko *, Mamta Sattavat ¶, Henry Sanchez ¶,
Ana Serban * , Martin Vey ||, Henry Baron **, Kurt Giles *,
, Bruce L. Miller , , Stephen J. DeArmond * , ¶ and Stanley
B. Prusiner *, , ,
*Institute for Neurodegenerative Diseases, Memory and Aging
Center, and Departments of Neurology, ¶Pathology, and
Biochemistry and Biophysics, University of California, San
Francisco, CA 94143; ||ZLB Behring, 35041 Marburg, Germany;
and **ZLB Behring, 75601 Paris, France
Contributed by Stanley B. Prusiner, December 22, 2004
Abstract
With the discovery of the prion protein (PrP),
immunodiagnostic procedures were applied to diagnose
Creutzfeldt–Jakob disease (CJD). Before development of the
conformation-dependent immunoassay (CDI), all immunoassays
for the disease-causing PrP isoform (PrPSc) used limited
proteolysis to digest the precursor cellular PrP (PrPC).
Because the CDI is the only immunoassay that measures both
the protease-resistant and protease-sensitive forms of
PrPSc, we used the CDI to diagnose human prion disease. The
CDI gave a positive signal for PrPSc in all 10–24 brain
regions (100%) examined from 28 CJD patients. A subset of 18
brain regions from 8 patients with sporadic CJD (sCJD) was
examined by histology, immunohistochemistry (IHC), and the
CDI. Three of the 18 regions (17%) were consistently
positive by histology and 4 of 18 (22%) by IHC for the 8
sCJD patients. In contrast, the CDI was positive in all 18
regions (100%) for all 8 sCJD patients. In both gray and
white matter, 90% of the total PrPSc was protease-sensitive
and, thus, would have been degraded by procedures using
proteases to eliminate PrPC. Our findings argue that the CDI
should be used to establish or rule out the diagnosis of
prion disease when a small number of samples is available as
is the case with brain biopsy. Moreover, IHC should not be
used as the standard against which all other
immunodiagnostic techniques are compared because an
immunoassay, such as the CDI, is substantially more
sensitive.
snip...
Discussion
The clinical diagnosis of human prion disease is often
difficult until the patient shows profound signs of
neurologic dysfunction. It is widely accepted that the
clinical diagnosis must be provisional until a tissue
diagnosis either confirms or rules out the clinical
assessment. Before the availability of Abs to PrP, a tissue
diagnosis was generally made by histologic evaluation of
neuropil vacuolation. IHC with
anti-glial-fibrillary-acidic-protein Abs in combination with
H&E staining preceded the use of anti-PrP Ab staining.
Recently, the role of IHC in the diagnosis of scrapie in the
brains of eight clinically affected goats inoculated with
the SSBP1 prion isolate has been challenged (14). Thalamic
samples taken from seven of eight goats with scrapie were
positive for PrPSc by Western blotting but negative by IHC.
The eighth goat was negative by Western blotting and IHC.
Consistent with these findings in goats are the data
reported here, in which IHC of formalin-fixed,
paraffin-embedded human brain samples was substantially less
sensitive than the CDI.
The CDI was developed to quantify PrPSc in tissue samples
from mammals producing prions. Concerned that limited PK
digestion was hydrolyzing some or even most of the PrPSc, we
developed a CDI that does not require PK digestion. The CDI
revealed that as much as 90% of PrPSc is sPrPSc; thus, it
was being destroyed during limited proteolytic digestion
used to hydrolyze PrPC. sPrPSc comprises 80% of PrPSc in the
frontal lobe and in the white matter (Fig. 4).
The CDI detected HuPrPSc with a sensitivity comparable to
the bioassay for prion infectivity in Tg(MHu2M) mice (Fig.
1). The high sensitivity achieved by the CDI is due to
several factors (8, 10, 11, 15). First, both sPrPSc and
rPrPSc conformers are specifically precipitated by PTA
(Table 5) (8, 9). PTA has also been used to increase the
sensitivity of Western blots enabling the detection of
rPrPSc in human muscle and other peripheral tissues (16,
17). Second, a sandwich protocol was used with the
high-affinity MAR1 mAb (11) to capture HuPrPSc and
Eu-labeled 3F4 mAb to detect HuPrPSc (12). Third, the CDI
detects PrPSc by Ab-binding to native and denatured forms of
the protein and, therefore, does not depend on proteolytic
degradation of PrPC. We chose not to perform Western blots
on most of the samples used in this study because such
immunoblots require denaturation of the sample, which
eliminates measurement of the native signal corresponding to
PrPC (Table 5). Moreover, a comparison between the CDI and
Western blotting on brain samples from sCJD and variant CJD
patients showed that the CDI was 50- to 100-fold more
sensitive (15). Additionally, Western blots combined with
densitometry are linear over a 10- to 100-fold range of
concentrations, whereas the CDI is linear over a >104-fold
range. The CDI has been automated, which not only improves
accuracy and reproducibility (10) but also allows numerous
samples to be analyzed, as reported here. Western blots are
difficult to automate and are labor intensive.
Our studies show that only the CDI detected PrPSc in all
regions examined in 24 sCJD and 3 fCJD(E200K) brains (Figs.
2 and 6). Comparative analyses demonstrated that the CDI was
vastly superior to histology and IHC. When 18 regions of 8
sCJD and 2 fCJD(E200K) brains were compared, we discovered
that histology and IHC were unreliable diagnostic tools
except for samples from a few brain regions. In contrast,
the CDI was a superb diagnostic procedure because it
detected PrPSc in all 18 regions in 8 of 8 sCJD and 2 of 2
fCJD(E200K) cases (Tables 1 and 2).
Histologic changes in prion disease have been shown to
follow the accumulation of prions as measured by bioassay of
infectivity and by PrPSc accumulation (18–22). Because low
levels of PrPSc are not associated with neuropathologic
changes, some discrepancy between vacuolation and PrPSc was
expected. In contrast to histology, IHC measures PrP
immunostaining after autoclaving tissue sections exposed to
formic acid. Because IHC measures PrP, we expected the
sensitivity of this procedure might be similar to the CDI,
but that proved not to be the case. Whether exposure of
formic acid-treated tissue sections to elevated temperature
destroys not only PrPC but also sPrPSc and only denatures
rPrPSc remains to be determined. Such a scenario could
account for the lower sensitivity of IHC compared with CDI
or bioassay (Tables 1 and 2).
Studies of the white matter in CJD brains were particularly
informative with respect to the sensitivity of the CDI,
where PrPSc levels were low but readily detectable, 10- to
100-fold above the threshold value (Fig. 4). Because animal
studies have shown that PrPSc and infectivity are
transported anterogradely from one brain region to another
along neuroanatomical pathways (23–25), we expected to find
PrPSc in white matter as demonstrated by the CDI but not
IHC. Axonal transport of PrPSc is also suggested by
diffusion-weighted MRI scans of CJD cases, which show
high-intensity signals in analogous neocortical regions of
the right and left cerebral hemispheres (26). This symmetry
of neuroradiological abnormalities is consistent with spread
of PrPSc to the contralateral cortex by means of callosal
commissural pathways.
Most immunoassays that detect HuPrPSc do so only after
subjecting the sample to limited proteolysis to form PrP
27–30, followed by denaturation. Because the CDI measures
the immunoreactivity before and after denaturation to an
epitope that is exposed in native PrPC but buried in PrPSc,
limited proteolysis to eliminate PrPC is unnecessary. Assays
based on limited proteolysis underestimate the level of
PrPSc because they digest sPrPSc, which represents 80–90% of
PrPSc in CJD and scrapie brains (Fig. 4 and Table 5).
Gerstmann–Sträussler–Scheinker, an inherited human prion
disease, is caused by the P102L mutation in the PRNP gene.
In mice expressing the Gerstmann–Sträussler–Scheinker mutant
PrP transgene, the CDI detected high levels of sPrPSc(P101L)
as well as low levels of rPrPSc(P101L) long before
neurodegeneration and clinical symptoms occurred (9).
sPrPSc(P101L) as well as low concentrations of rPrPSc(P101L)
previously escaped detection (27). Whether a similar
situation applies in other genetic forms of prion disease,
sCJD, or variant CJD remains to be determined. Because most
of the PrPSc in the brains of sCJD patients is
protease-sensitive (Fig. 4), it is likely that the lower
sensitivity of IHC is due to its inability to detect sPrPSc.
Presently, we have no information about the kinetics of
either sPrPSc or rPrPSc accumulation in human brain. Limited
information on the kinetics of PrPSc accumulation in
livestock comes from studies of cattle, sheep, and goats
inoculated orally, but most of the bioassays were performed
in non-Tg mice (28–30) in which prion titers were
underestimated by as much as a factor of 104 (10).
The studies reported here are likely to change profoundly
the approach to the diagnosis of prion disease in both
humans and livestock (31–33). The superior performance of
the CDI in diagnosing prion disease compared to routine
neuropathologic examination and IHC demands that the CDI be
used in future diagnostic evaluations of prion disease.
Prion disease can no longer be ruled out by routine
histology or IHC. Moreover, the use of IHC to confirm cases
of bovine spongiform encephalopathy after detection of
bovine PrPSc by the CDI (10) seems an untenable approach in
the future. Clearly, the CDI for HuPrPSc is as sensitive or
more sensitive than bioassays in Tg(MHu2M) mice (Fig. 1).
Our results suggest that using the CDI to test large numbers
of samples for human prions might alter the epidemiology of
prion diseases. At present, there is limited data on the
frequency of subclinical variant CJD infections in the U.K.
population (34). Because appendixes and tonsils were
evaluated only by IHC, many cases might have escaped
detection (Tables 1 and 2). Equally important may be the use
of CDI-like tests to diagnose other neurodegenerative
disorders, such as Alzheimer's disease, Parkinson's disease,
and the frontotemporal dementias. Whether IHC underestimates
the incidence of one or more of these common degenerative
diseases is unknown. Moreover, CDI-like tests may help
determine the frequency with which these disorders and the
prion diseases occurs concomitantly in a single patient (35,
36).
Acknowledgements
snip...END
http://www.pnas.org/
Volume 349:1812-1820 November 6, 2003 Number 19
Extraneural Pathologic Prion Protein in Sporadic Creutzfeldt-Jakob Disease
Background In patients with sporadic Creutzfeldt–Jakob disease, pathologic disease-associated prion protein (PrPSc) has been identified only in the central nervous system and olfactory-nerve tissue. Understanding the distribution of PrPSc in Creutzfeldt–Jakob disease is important for classification and diagnosis and perhaps even for prevention.
Methods We used a highly sensitive method of detection — involving the concentration of PrPSc by differential precipitation with sodium phosphotungstic acid, which increased the sensitivity of Western blot analysis by up to three orders of magnitude — to search for PrPSc in extraneural organs of 36 patients with sporadic Creutzfeldt–Jakob disease who died between 1996 and 2002.
Results PrPSc was present in the brain tissue of all patients. In addition, we found PrPSc in 10 of 28 spleen specimens and in 8 of 32 skeletal-muscle samples. Three patients had PrPSc in both spleen and muscle specimens. Patients with extraneural PrPSc had a significantly longer duration of disease and were more likely to have uncommon molecular variants of sporadic Creutzfeldt–Jakob disease than were patients without extraneural PrPSc.
Conclusions Using sensitive techniques, we identified extraneural deposition of PrPSc in spleen and muscle samples from approximately one third of patients who died with sporadic Creutzfeldt–Jakob disease. Extraneural PrPSc appears to correlate with a long duration of disease.
Source Information
From the Institute of Neuropathology and National Reference Center for Prion Diseases, University Hospital of Zurich, Zurich, Switzerland.
Dr. Glatzel and Mr. Abela contributed equally to the article.
Address reprint requests to Dr. Aguzzi at the Institute of Neuropathology, University Hospital of Zurich, Schmelzbergstr. 12, CH-8091 Zurich, Switzerland, or at [email protected] .
http://content.nejm.org/cgi/
Creutzfeldt-Jakob disease and inclusion body myositis: Abundant disease-associated prion protein in muscle
Gabor G. Kovacs, MD PhD 1 2, Elisabeth Lindeck-Pozza, MD 1, Leila Chimelli, MD, PhD 3, Abelardo Q. C. Araújo, MD, PhD 4, Alberto A. Gabbai, MD, PhD 5, Thomas Ströbel, PhD 1, Markus Glatzel, MD 6, Adriano Aguzzi, MD, PhD 6, Herbert Budka, MD 1 *
1Institute of Neurology, University of Vienna, and Austrian Reference Centre for Human Prion Diseases, Vienna, Austria
2National Institute of Psychiatry and Neurology, Budapest, Hungary
3Department of Pathology, School of Medicine, Federal University of Rio de Janeiro
4Department of Neurology, School of Medicine, Federal University of Rio de Janeiro
5Department of Neurology, School of Medicine, Federal University of Sao Paulo, Brazil
6Institute of Neuropathology, University Hospital of Zürich, Zürich, Switzerland
email: Herbert Budka ([email protected]. )
*Correspondence to Herbert Budka, Institute of Neurology, AKH 4J, Wühringer Gürtel 18-20, POB 48, A-1097 Vienna, Austria
Funded by:
European Union (EU) Project; Grant Number: TSELAB QLK2-CT-2002-81523
EU Concerted Action PRIONET; Grant Number: QLK2-2000-CT-00837
Abstract
Pathologicalprion protein (PrPSc) is the hallmark of prion diseases affecting primarily the central nervous system. Using immunohistochemistry, paraffin-embedded tissue blot, and Western blot, we demonstrated abundant PrPSc in the muscle of a patient with sporadic Creutzfeldt-Jakob disease and inclusion body myositis. Extraneural PrPC-PrPSc conversion in Creutzfeldt-Jakob disease appears to become prominent when PrPC is abundantly available as substrate, as in inclusion body myositis muscle.
--------------
Received: 16 June 2003; Revised: 11 September 2003; Accepted: 11 September 2003
Digital Object Identifier (DOI)
10.1002/ana.10813 About DOI
http://www3.interscience.wiley.com/
AS Professor Aguzzi kindly put it most recently ;
107
Vet Pathol 42:107 108 (2005)
Letters to the Editor
Editor:
Absence of evidence is not always evidence of absence. In the article Failure to detect prion protein (PrPres) by immunohistochemistry in striated muscle tissues of animals experimentally inoculated with agents of transmissible spongiform encephalopathy, recently published in Veterinary Pathology (41:78 81, 2004), PrPres was not detected in striated muscle of experimentally infected elk, cattle, sheep, and raccoons by immunohistochemistry (IHC). Negative IHC, however, does not exclude the presence of PrPSc. For example, PrPres was detected in skeletal muscle in 8 of 32 humans with the prion disease, sporadic Creutzfeldt-Jakob disease (CJD), using sodium phosphotungstic acid (NaPTA) precipitation and western blot.1 The NaPTA precipitation, described by Wadsworth et al.,3 concentrates the abnormal isoform of the prion, PrPres, from a large tissue homogenate volume before western blotting. This technique has increased the sensitivity of the western blot up to three orders of magnitude and could be included in assays to detect PrPres. Extremely conspicuous deposits of PrPres in muscle were detected by IHC in a recent case report of an individual with inclusion body myositis and CJD.2 Here, PrPres was detected in the muscle by immunoblotting, IHC, and paraf- fin-embedded tissue blot. We would therefore caution that, in addition to IHC, highly sensitive biochemical assays and bioassays of muscle are needed to assess the presence or absence of prions from muscle in experimental and natural TSE cases.
Christina Sigurdson, Markus Glatzel, and Adriano Aguzzi
Institute of Neuropathology
University Hospital of Zurich
Zurich, Switzerland
References
1 Glatzel M, Abela E, et al: Extraneural pathologic prion
protein in sporadic Creutzfeldt-Jakob disease. N Engl J
Med 349(19):1812 1820, 2003
2 Kovacs GG, Lindeck-Pozza E, et al: Creutzfeldt-Jakob
disease and inclusion body myositis: abundant diseaseassociated
prion protein in muscle. Ann Neurol 55(1):
121 125, 2004
3 Wadsworth JDF, Joiner S, et al: Tissue distribution of protease
resistant prion protein in variant CJD using a highly
sensitive immuno-blotting assay. Lancet 358:171 180,
2001...///
EMBO reports AOP Published online: 11 April 2003
Widespread PrPSc accumulation in muscles of hamsters orally infected with scrapie
http://www.emboreports.org/
Watson Meldrum et al Scrapie agent in muscle - Pattison I A (1990)
Veterinary record, 20 January 1990. p.68
http://www.bseinquiry.gov.uk/files/yb/1990/01/19009001.pdf
===============================
GREETINGS AGAIN APHIS ET AL,
FURTHERMORE, WE HAVE FAILED TO EVEN STOP THE SRMs FROM WHOLE
CUTS OF BONELESS BEEF IMPORTED FROM CANADA IN THE VERY ONSET OF
THE NEW BSE MRR (MINIMAL RISK REGION). THIS IS THE VERY REASON I HAVE
SAID TIME AND TIME AGAIN THAT BY THIS ADMINISTRATION ABANDONING THE
BSE GBR RISK ASSESSMENTS BECAUSE THEY DID NOT LIKE THE ASSESSMENT
OF BSE GBR III, AND ADHERING TO A NEW BSE MRR POLICY THAT WAS DESIGNED
NOT FOR HUMAN HEALTH, BUT ONLY FOR COMMODITIES AND FUTURES, WILL FURTHER
EXPOSE NEEDLESSLY MILLIONS AND MILLIONS OF HUMANS AND ANIMALS VIA THE FREE
TRADING OF ALL STRAINS OF TSE GLOBALLY. references as follow ;
Wisconsin Firm Recalls Beef Products
Recall Release CLASS II RECALL
FSIS-RC-032-2005 HEALTH RISK: LOW
Congressional and Public Affairs
(202) 720-9113
Steven Cohen
WASHINGTON, Aug. 19, 2005 - Green Bay Dressed Beef, a Green Bay, Wis., establishment, is voluntarily recalling approximately 1,856 pounds of beef products that may contain portions of the backbone from a cow just over 30 months old, the U.S. Department of Agriculture's Food Safety and Inspection Service announced today. The product was from a cow imported directly for slaughter from Canada.
Based on information provided by Canada, the products subject to this Class II recall are from a cow that is approximately one month older than the 30-month age limit. Both ante-mortem and post-mortem inspection were done on the cow in question. FSIS inspection program personnel determined the cow to be healthy and fit for human food. FSIS' designation of this recall as Class II is because it is a situation where there is a remote probability of adverse health consequences from the use of the product.
FSIS learned about this as a result of a Canadian audit of their health certificate that accompanied the imported cow. Prior to slaughter, the health certificate accompanying the cow was presented to the establishment, and it appeared complete and accurate. However, a subsequent audit of information related to the health certificate by Canadian officials found that it was not accurate. Action has been taken by Canadian Food Inspection Agency officials in response to findings from the audit.
The products subject to recall are:
Five boxes of 243 lb. vacuum pouched packages of "American Foods Group, NECKBONE UNTRIM'D, USDA CHOICE OR HIGHER" with the case code of 77333;
One box of 50 lb. vacuum pouched package of "American Foods Group, SHORTLOIN 2X2, USDA SELECT OR HIGHER" with the case code of 75231;
One box of 60 lb. vacuum pouched package of "American Foods Group, SHORTLOIN 2X2, USDA CHOICE OR HIGHER" with the case code of 75060;
Five boxes of 258 lb. vacuum pouched packages of "Dakota Supreme Beef, SHORTLOIN 0X11/4, USDA SELECT OR HIGHER" with the case code of 75442;
Sixteen boxes of 811 lb. vacuum pouched packages of "American Foods Group, BLADE BI N/O CHUCK, USDA CHOICE OR HIGHER" with the case code of 75955;
Nine boxes of 435 lb. vacuum pouched packages of "American Foods Group, BLADE BI N/O CHUCK, USDA SELECT OR HIGHER" with the case code of 75952.
Each box bears the establishment number "410" inside the USDA seal of inspection. The products were produced on August 4, and were distributed to wholesale distributors in Pennsylvania, Florida, Illinois, Maryland, Minnesota and Wisconsin.
Under the interim final rules FSIS implemented on January 12, 2004, certain specified risk materials must be removed from all cattle depending on the age of the animal. On this animal all specified risk materials for cattle 30 months and over were removed, with the exception of the vertebral column. At the time of slaughter, the animal was certified to be under 30 months of age and removal of the vertebral column was not required. A subsequent audit determined the animal was just over 30 months of age; therefore, the vertebral column is required to be removed. This is the reason for the recall of the selected products.
Consumers with questions about the recall may contact Sally VandeHei, Executive Assistant at 1-877-894-3927. National media with questions may contact Jim Mulhern at (202) 496-2468. Local media with questions may contact Susan Finco at (920) 965-7750 ext.158.
Consumers with other food safety questions can phone the toll-free USDA Meat and Poultry Hotline at 1-888-MPHotline (1-888-674-6854). The hotline is available in English and Spanish and can be reached from 10 a.m. to 4 p.m. (Eastern Time), Monday through Friday. Recorded food safety messages are available 24 hours a day.
Sample Product Labels: These are similar to, but not identical to, labels on the recalled product.
#
USDA Recall Classifications
Class I This is a health hazard situation where there is a reasonable probability that the use of the product will cause serious, adverse health consequences or death.
Class II This is a health hazard situation where there is a remote probability of adverse health consequences from the use of the product.
Class III This is a situation where the use of the product will not cause adverse health consequences.
http://www.fsis.usda.gov/News_&_Events/Recall_032_2005_Release/index.asp
:
Docket No. 03-080-1 -- USDA ISSUES PROPOSED RULE TO ALLOW LIVE ANIMAL IMPORTS FROM CANADA
[TSS SUBMISSION 11/03/2003 01:19 PM To: [email protected] ]
https://web01.aphis.usda.gov/BSEcom.nsf/0/b78ba677e2b0c12185256dd300649f9d?OpenDocument&AutoFramed
THE BSE MRR POLICY SHOULD BE ABOLISHED/REPEALED IMMEDIATELY AND THE BSE GBR RISK ASSESSMENTS AND POLICY SHOULD BE STRICTLY ENFORCED AND FURTHER ENHANCED TO INCLUDE CWD AND ALL TSEs...TSS
EFSA Scientific Report on the Assessment of the Geographical BSE-Risk (GBR) of the United States of America (USA)
Publication date: 20 August 2004
Adopted July 2004 (Question N° EFSA-Q-2003-083)
Report
Summary
Summary of the Scientific Report
The European Food Safety Authority and its Scientific Expert Working Group on the Assessment of the Geographical Bovine Spongiform Encephalopathy (BSE) Risk (GBR) were asked by the European Commission (EC) to provide an up-to-date scientific report on the GBR in the United States of America, i.e. the likelihood of the presence of one or more cattle being infected with BSE, pre-clinically as well as clinically, in USA. This scientific report addresses the GBR of USA as assessed in 2004 based on data covering the period 1980-2003.
The BSE agent was probably imported into USA and could have reached domestic cattle in the middle of the eighties. These cattle imported in the mid eighties could have been rendered in the late eighties and therefore led to an internal challenge in the early nineties. It is possible that imported meat and bone meal (MBM) into the USA reached domestic cattle and leads to an internal challenge in the early nineties.
A processing risk developed in the late 80s/early 90s when cattle imports from BSE risk countries were slaughtered or died and were processed (partly) into feed, together with some imports of MBM. This risk continued to exist, and grew significantly in the mid 90's when domestic cattle, infected by imported MBM, reached processing. Given the low stability of the system, the risk increased over the years with continued imports of cattle and MBM from BSE risk countries.
EFSA concludes that the current GBR level of USA is III, i.e. it is likely but not confirmed that domestic cattle are (clinically or pre-clinically) infected with the BSE-agent. As long as there are no significant changes in rendering or feeding, the stability remains extremely/very unstable. Thus, the probability of cattle to be (pre-clinically or clinically) infected with the BSE-agent persistently increases.
http://www.efsa.eu.int/science/efsa_scientific_reports/gbr_assessments/573_en.html
From: Terry S. Singeltary Sr. [[email protected]]
Sent: Tuesday, July 29, 2003 1:03 PM
To: [email protected]
Cc: [email protected]; [email protected].; BSE-L
Subject: Docket No. 2003N-0312 Animal Feed Safety System [TSS SUBMISSION
TO DOCKET 2003N-0312]
Greetings FDA,
snip...
PLUS, if the USA continues to flagrantly ignore the _documented_ science to date about the known TSEs in the USA (let alone the undocumented TSEs in cattle), it is my opinion, every other Country that is dealing with BSE/TSE should boycott the USA and demand that the SSC reclassify the USA BSE GBR II risk assessment to BSE/TSE GBR III 'IMMEDIATELY'. for the SSC to _flounder_ any longer on this issue, should also be regarded with great suspicion as well. NOT to leave out the OIE and it's terribly flawed system of disease surveillance. the OIE should make a move on CWD in the USA, and make a risk assessment on this as a threat to human health. the OIE should also change the mathematical formula for testing of disease. this (in my opinion and others) is terribly flawed as well. to think that a sample survey of 400 or so cattle in a population of 100 million, to think this will find anything, especially after seeing how many TSE tests it took Italy and other Countries to find 1 case of BSE (1 million rapid TSE test in less than 2 years, to find 102 BSE cases), should be proof enough to make drastic changes of this system. the OIE criteria for BSE Country classification and it's interpretation is very problematic. a text that is suppose to give guidelines, but is not understandable, cannot be considered satisfactory. the OIE told me 2 years ago that they were concerned with CWD, but said any changes might take years. well, two years have come and gone, and no change in relations with CWD as a human health risk. if we wait for politics and science to finally make this connection, we very well may die before any decisions
or changes are made. this is not acceptable. we must take the politics and the industry out of any final decisions of the Scientific community. this has been the problem from day one with this environmental man made death sentence. some of you may think i am exaggerating, but you only have to see it once, you only have to watch a loved one die from this one time, and you will never forget, OR forgive...yes, i am still very angry... but the transmission studies DO NOT lie, only the politicians and the industry do... and they are still lying to this day...TSS
http://www.fda.gov/ohrms/dockets/dockets/03n0312/03N-0312_emc-000001.txt
GREETINGS AGAIN APHIS ET AL,
Moving on to the theory that BSE agent is not in blood. THIS is what they use to think with nvCJD. However
the nvCJD agent has now been detected and transmitted the TSE agent by blood. nvCJD is the BSE agent
that has transmitted to humans. nvCJD is human BSE. so if nvCJD transmits by blood, why not BSE? with
the limited testing to date, the limited sensitivity of the detection of the BSE/TSE agent blood to date, i would
not be so sure that the BSE/TSE agent does not transmit by blood. just recent Ag. Comm. Johanns stated
that they would not address the blood issue being fed to cattle. a foolish and careless mistake. but typical.
we now have detected new atypical strains of the BSE/TSE agent in cattle in many countries i.e. Japan, France,
Belgium, Germany, and Italy. In the Italian study of BASE, a new? TSE in cattle they have discovered that is
not like the nvCJD, but very similar to sporadic CJD. They have detected 2 such cattle at printing of this study
March 2, 2004. Identification of a second bovine amyloidotic spongiform encephalopathy: Molecular similarities
with sporadic Creutzfeldt-Jakob disease PNAS. MY point is that with these new atypical TSEs showing up in
cattle, sheep and goats, we don't know if blood and other tissues transmits the disease. THE SRM list may
and should be reevaluated. WE know BSE is in the USA, but we also know that in the studies of Mission Texas,
where USA suffolk scrapie sheep were inoculated into USA cattle, the PRION agent that was produced did not
look like the UK BSE strain. so why would all CJD cases in the USA look like the UK human BSE i.e. nv/v CJD?
WHY wait and expose millions and millions needlessly as in the past with previous TSE blunders, why wait to
act. why not act first with what we know, which is very concerning, then as science evolves, go from there.
references as follow ;
UK Strategy for Research and
Development on Human and Animal
Health Aspects of Transmissible
Spongiform Encephalopathies
2005-2008
snip...
3.2 Tackling the spread of infection
3.2.1 The species barrier and the carrier state
3.2.1.1 The possibility of 'carrier' states in animals and humans, and our present inability
to identify them, pose a potential threat to public and animal health. The susceptibility of
humans to BSE infection, and the ability of the disease to remain clinically silent for
many years, is of major concern to DH. Although the death of a UK blood donor from
vCJD in 1999 three years after making the donation and the subsequent death from vCJD
of the recipient in 2003 have not been causally linked, transmission of infection through
blood transfusion is the most likely explanation327. The case heightens concerns that
'carriers' could be transmitting the disease through blood, tissue and organ donation or
by contaminating surgical instruments when undergoing surgery. DH will continue to
support research to detect infectious prions in human tissue, to investigate the
decontamination of surgical instruments and to develop measures to protect blood
supplies.
3.2.1.2 Animal models of some TSEs have detected infectivity in blood. Experiments,
which have involved transfusing large volumes of blood from infected sheep to healthy
recipient sheep, have demonstrated that infectivity can be transmitted by blood
transfusion. A central part of DH policy in this area has been the leucodepletion of blood
donations and the efficacy of this technology can now be tested in sheep.
snip...
4.5.4 In 1987, epidemiological studies of BSE cases identified meat and bone meal as the
probable means by which the disease was being spread. In an attempt to prevent
further infections a ban on incorporating ruminant protein in ruminant feed was
introduced in July 1988. Due to the long incubation period associated with this disease
the efficacy of this control measure was not immediately apparent. As time passed it
became clear from the number of cases born after the ban that it was not wholly
effective.
4.5.5 Epidemiological analysis of these cases showed that a high proportion of them
occurred in areas where the pig population was high. This observation, coupled with
research data that showed that only a very small dose of the infective material was
needed to cause disease in cattle, led to the conclusion that cross-contamination of feed
was occurring.
4.5.6 Since 1988, increasingly stringent feed controls have been put in place. Key
amongst these have been:
• the ban on the use of specified bovine offal in all animal feed (September 1990);
• the ban on feeding any farmed animal, including horses and fish, with mammalian
meat and bone meal. (This began in March 1996 but following this ban there was
a recall scheme and the date from which the ban was considered to be fully
effective is regarded as being 1 August 1996);
62
v6.1
• EU-wide controls on feed which extended the ban to include all processed animal
protein, including that derived from birds and fish (implemented in the UK from 1
August 2001).
4.5.7 As illustrated in fig. [ ] these later measures have reduced the spread of BSE.
However, they have not been one hundred per cent effective. As at 31st December 2003
there had been 81 cases of BSE in animals born since 1 August 1996 in the UK.
full text 91 pages;
http://www.mrc.ac.uk/pdf-about-tse_uk_strategy_june2005.pdf
GREETINGS AGAIN APHIS ET AL,
JUST what about those old studies at Mission Texas and the atypical TSE in cattle?
would it not be prudent for human health purpose, the question that, with all the atypical
TSEs showing up in animal and man in different countries, the fact that most all of these
TSEs transmit as freely or not as freely as BSE (depending whom you have witnessed die from
this agent either directly or indirectly via a multitude of potential routes and sources) to primates.
would it not be prudent to ask yourself if some if not all of these sporadic CJDs might be a
by-product of these TSEs either directly or indirectly via a multitude of proven routes and sources
in animal studies? it is unethical for human transmission studies considering the fact that the agent
is 100% fatal, slow, but fatal. there has been no sound science in any of the recent decisions in the
USA in regards to BSE/TSE human or animal, all one has to do is look at TEXAS, the mad cow that
got away, the stumbling and staggering one that NO TSE TEST AT ALL was done, ordered rendered,
and then the infamous positive, positve, secret positive, inconclusive (NO WB), negative, 8 month delay,
then the 'Fong Effect' took place, THEN FINALLY CONFIRMED SOME 8 MONTHS LATER IN WEYBRIDGE.
Finally recently, another inconclusive that took place that sat untested on some shelf for about 4 months while
the Texas mad cow blunder was going on. the tissues of this cow this time were preserved in preservative as
to render any WB for further confirmation, what i called the 'FONG SYNDROME' or the 'end around' the WB
ordered previously by the Honorable Phyllis Fong of the OIG. Politics at its finest, to hell with human health.
WE find now that 9,200 USA POTENTIAL MAD COWS IN JUNE 2004 ENHANCED COVER-UP
SURVEILLANCE PROGRAM WENT UNTESTED WITH NO RAPID TEST OR WB, ONLY IHC, the test
that fails the most, that is very unreliable as noted above in my submission by Prusiner et al and other scientist.
NOW, back to Mission, Texas ;
>> Differences in tissue distribution could require new regulations
>> regarding specific risk material (SRM) removal.
snip...end
full text 33 PAGES ;
http://www.bseinquiry.gov.uk/files/mb/m11b/tab01.pdf
http://www.bseinquiry.gov.uk/files/yb/1988/10/00001001.pdf
It was, however, performed in the USA in 1979, when it was shown that cattle
inoculated with the scrapie agent endemic in the flock of Suffolk sheep at
the United States Department of Agriculture in Mission, Texas, developed a
TSE quite unlike BSE. 32 The findings of the initial transmission, though
not of the clinical or neurohistological examination, were communicated in
October 1988 to Dr Watson, Director of the CVL, following a visit by Dr
Wrathall, one of the project leaders in the Pathology Department of the CVL,
to the United States Department of Agriculture. 33 The results were not
published at this point, since the attempted transmission to mice from the
experimental cow brain had been inconclusive. The results of the clinical
and histological differences between scrapie-affected sheep and cattle were
published in 1995. Similar studies in which cattle were inoculated
intracerebrally with scrapie inocula derived from a number of
scrapie-affected sheep of different breeds and from different States, were
carried out at the US National Animal Disease Centre. 34 The results,
published in 1994, showed that this source of scrapie agent, though
pathogenic for cattle, did not produce the same clinical signs of brain
lesions characteristic of BSE.
http://www.bseinquiry.gov.uk/
1: J Infect Dis. 1994 Apr;169(4):814-20.
Intracerebral transmission of scrapie to cattle.
Cutlip RC, Miller JM, Race RE, Jenny AL, Katz JB, Lehmkuhl HD, DeBey BM,
Robinson MM.
USDA, Agriculture Research Service, National Animal Disease Center, Ames, IA
50010.
To determine if sheep scrapie agent(s) in the United States would induce a
disease in cattle resembling bovine spongiform encephalopathy, 18 newborn
calves were inoculated intracerebrally with a pooled suspension of brain
from 9 sheep with scrapie. Half of the calves were euthanatized 1 year after
inoculation. All calves kept longer than 1 year became severely lethargic
and demonstrated clinical signs of motor neuron dysfunction that were
manifest as progressive stiffness, posterior paresis, general weakness, and
permanent recumbency. The incubation period was 14-18 months, and the
clinical course was 1-5 months. The brain from each calf was examined for
lesions and for protease-resistant prion protein. Lesions were subtle, but a
disease-specific isoform of the prion protein was present in the brain of
all calves. Neither signs nor lesions were characteristic of those for
bovine spongiform encephalopathy.
MeSH Terms:
Animals
Brain/microbiology*
Brain/pathology
Cattle
Cattle Diseases/etiology*
Cattle Diseases/pathology
Encephalopathy, Bovine Spongiform/etiology*
Encephalopathy, Bovine Spongiform/pathology
Immunoblotting/veterinary
Immunohistochemistry
Male
Motor Neurons/physiology
Prions/analysis
Scrapie/pathology
Scrapie/transmission*
Sheep
Sleep Stages
Time Factors
Substances:
Prions
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8133096&dopt=Citation
Intracerebral transmission of scrapie to cattle FULL TEXT PDF;
SNIP...
Discussion
WE conclude that American sources of sheep scrapie are transmissible to
cattle by direct intracerebral inoculation but the disease induced is NOT
identical to BSE as seen in the United Kingdom. While there were
similarities in clinical signs between this experimental disease and BSE,
there was no evidence of a
