Brad S said:
While I'm aware of the lone incedent in Japan where an animal was perported to be 21 months old and diagnosed with bse, yes I'm saying the test is claimed by noone to be effective on the vast majority of US raised fed beef. I realize that the lone savior to "testing fed beef" is on an animal perported to gain less than 2# per day of age. That ain't reality in the US or Canada.
I've turned trip after trip born in march and processed in feb(11 months later). This is along the lines of what Creekstone processes. OT, What test claims to be able to test for bse in these cattle?
Young steer negative for BSE
20-month-old animal would have been one of youngest to have disease
TOKYO (AP)--A 20-month-old steer in northeastern Japan initially suspected of having bovine spongiform encephalopathy has tested negative, a health official said April 18.
The young Holstein was slaughtered for meat recently in Fukushima prefecture (state) and initially tested positive for the brain-wasting disease.
But further tests at the National Institute of Infectious Diseases in Tokyo showed the steer did not have BSE, according to prefectural health official Shinichi Nakajima.
The case had caused concern because of the steer's young age. The youngest cow to have contracted the disease as of 2005 was a 20-month-old cow found in the United Kingdom in 1992, according to Japan's Food Safety Commission.
Separately, inspectors in Okayama prefecture (state), west of Tokyo, found late April 17 that a 6-year-old dairy cow, intended to be slaughtered for meat, has tested positive for the disease, said health official Waichiro Kawai.
Confirmation of the results could come as early as April 19, Kawai said.
Japan in December eased a two-year-old ban on U.S. beef to allow imports from cows aged 20 months or younger which did not contain body parts thought at risk of the brain-wasting disease. The trade was again halted after a U.S. veal shipment was found to contain prohibited bones.
The United States, in contrast, requires removal of at-risk parts from animals older than 30 months, although there is a short list of tissues that must be removed from younger animals. U.S. officials have argued cows younger than that face minuscule risks of BSE.
Japan's ban on U.S. beef has been detrimental to a trading relationship worth millions of dollars to American producers. Japan's market was worth US$1.4 billion annually when its government banned American beef in response to the first U.S. case of BSE in December 2003.
The two countries are still negotiating possible safeguards that might allow trade to resume.
BSE is a degenerative nerve disease in cattle. Eating contaminated meat products has been linked to the rare but fatal variant Creutzfeldt-Jakob disease.
Japan, which tests all cattle killed for meat, has confirmed 24 cases of BSE since 2001 - including three cases this year, according to the Agricultural Ministry. There have been three confirmed cases of the disease in the U.S.
5/1/06
http://www.hpj.com/archives/2006/may06/may1/YoungsteernegativeforBSE.cfm
BSE: Statistics - Youngest and oldest cases by year of onset (for passive surveillance cases) and on year of slaughter (for active surveillance cases)
As at 1 March 2007.
NB The last case in an animal aged 30 months or less was in 1996
Year of onset Age youngest case
(mnths) Age 2nd youngest case
(mnths) Age 2nd oldest case
(yrs.mnths) Age oldest case
(yrs.mnths)
1986 30 33 5.03 5.07
1987 30 31 9.09 10
1988 24 27 10.02 11.01 (2)
1989 21 24 (4) 12 (2) 15.04
1990 24 (2) 26 13.03 14
1991 24 26 (3) 14.02 17.05
1992 20 26 15.02 16.02
1993 29 30 (3) 14.1 18.1
1994 30 (2) 31 (2) 14.05 16.07
1995 24 32 14.09 15.05
1996 29 30 15.07 17.02
snip...
http://www.defra.gov.uk/animalh/bse/statistics/bse/yng-old.html
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 controversies
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.
T. Kitamoto (Ed.)PRIONSFood 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
1. 8. 2003.10.6. Fukushima Tochigi 2001.10.13. 23
2. 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.
Page 6 of 98
8/3/2006
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]
=================================
Atypical Proteinase K-Resistant Prion Protein (PrPres) observed in an Apparently Healthy 23-Month-Old Holstein Steer
Jpn. J. Infect. Dis., 56, 221-222, 2003
Laboratory and Epidemiology Communications
Atypical Proteinase K-Resistant Prion Protein (PrPres) Observed in an Apparently Healthy 23-Month-Old Holstein Steer
Yoshio Yamakawa*, KenÕichi Hagiwara, Kyoko Nohtomi, Yuko Nakamura, Masahiro Nishizima ,Yoshimi Higuchi1, Yuko Sato1,
Tetsutaro Sata1 and the Expert Committee for BSE Diagnosis, Ministry of Health, Labour and Welfare of Japan2
Department of Biochemistry & Cell Biology and 1Department of Pathology, National Institute of Infectious Diseases, Tokyo 162-
8640 and 2Miistry of Health, Labour and Welfare, Tokyo 100-8916
Communicated by Tetsutaro Sata
(Accepted December 2, 2003)
*Corresponding author: Mailing address: Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases,
Toyama 1-23-1, Shinjuku-ku, Tokyo 1628640, Japan. Tel: +81-3-5285-1111, Fax: +81-3-5285-1157, E-mail:
[email protected]
Since October 18, 2001, 'bovine spongiform encephalopathy (BSE) examination for all cattle slaughtered at abattoirs in the
country' has been mandated in Japan by the Ministry of Health, Labour and Welfare (MHLW). 'Plateria' ELISA-kit (Bio-Rad
Laboratories, Hercules, Calif., USA) is routinely used at abattoirs for detecting proteinase K (PK)-resistant prion protein (PrPSc) in
the obex region. Samples positive according to the ELISA screening are further subjected to Western blot (WB) and histologic and
immunohistochemical examination (IHC) at the National Institute of Infectious Diseases (NIID) or Obihiro University. If PrPSc is
detected either by WB or by IHC, the cattle are diagnosed as BSE. The diagnosis is approved by the Expert Committee for BSE
Diagnosis, MHLW. From October 18, 2001 to September 30, 2003, approximately 2.5 million cattle were screened at abattoirs. A
hundred and ten specimens positive according to ELISA were subjected to WB/IHC. Seven showed positive by both WB and IHC,
all exhibiting the typical electrophoretic profile of a high content of the di-glycosylated molecular form of PrPSc (1-3) and the
distinctive granular deposition of PrPSc in neuronal cells and neuropil of the dorsal nucleus of vagus.
An ELISA-positive specimen from a 23 month-old Holstein steer slaughtered on September 29, 2003, in Ibaraki Prefecture (Ibaraki
case) was sent to the NIID for confirmation. The animal was reportedly healthy before slaughter. The OD titer in ELISA was
slightly higher than the 'cut-off' level given by the manufacturer. The histology showed no spongiform changes and IHC revealed
no signal of PrPSc accumulation typical for BSE. However, WB analysis of the homogenate that was prepared from the obex
region and used for ELISA revealed a small amount of PrPSc with an electrophoretic profile different from that of typical BSEassociated
PrPSc (1-3). The characteristics were (i) low content of the di-glycosylated molecular form of PrPSc, (ii) a faster
migration of the non-glycosylated form of PrPSc on SDS-PAGE, and (iii) less resistance against PK digestion as compared with
an authentic PrPSc specimen derived from an 83-month-old Holstein (Wakayama case) (Fig. 1). Table 1 summarizes the relative
amounts of three distinctive glycoforms (di-, mono, non-glycosylated) of PrPSc calculated by densitometric analysis of the blot
shown in Fig. 1. As 2.5 mg wet weight obex-equivalent homogenate of the Ibaraki case (Fig. 1, lane 4) gave slightly stronger band
intensities of PrPSc than an 8 mg wet weight obex-equivqlent homogenate of a typical BSE-affected Wakayama case (Fig. 1, lane
2), the amount of PrPSc accumulated in the Ibaraki case was calculated to be 1/500 - 1/1000 of the Wakayama case. In the
Ibaraki case, the PrPSc bands were not detectable in the homogenates of the proximal surrounding region of the obex. These
findings were consistent with the low OD value in ELISA, i.e., 0.2 -0.3 for the Ibaraki case versus over 3.0 for the Wakayama case.
The DNA sequence of the PrP coding region of the Ibaraki case was the same as that appearing in the database (GenBank
accession number: AJ298878). More recently, we encountered another case that resembled the Ibaraki case. It was a 21-
monthold Holstein steer from Hiroshima Prefecture. WB showed typical BSE-specific PrPSc deposition though IHC did not detect
positive signals of PrPSc (data not shown).
Page 7 of 98
8/3/2006
Though the clinical onset of BSE is usually at around 5 years of age or later, a 20-month-old case showing the clinical signs has
been reported (4). Variant forms of BSE similar to our cases, i.e., with atypical histopathological and/or biochemical phenotype,
have been recently reported in Italy (5) and in France (6). Such variant BSE was not associated with mutations in the prion protein
(PrP) coding region as in our case (5,6).
The Ministry of Agriculture, Forestry and Fisheries of Japan (MAFF) announced a ban of feeding ruminants with meat bone meal
(MBM) on September 18, 2001, and a complete ban was made on October 15 of the same year. According to the recent MAFF
report, the previous seven cases of BSE in Japan were cattle born in 1995 - 1996 and possibly fed with cross-contaminated feed.
However, the two cattle in this report were born after the complete ban. Whether contaminated MBM was implicated in the present
cases remains to be investigated.
REFERENCES
Collinge, J., Sidle, K. C. L., Meads, J., Ironside, J. and Hill, A. F. (1996): Molecular analysis of prion strain variation and the
aetiology of 'new variant' CJD. Nature, 383, 685
690. Bruce, M. E., Will, R. G., Ironside, J. W., McConnell, I., Drummond, D., Suttie, A., McCardle, L., Chree, A., Hope, J., Birkett,
C., Cousens, S., Fraser, H. and Bostock, C. J. (1997): Transmissions to mice indicate that 'new variant' CJD is caused by the BSE
agent. Nature, 389, 498-501. Hill, A. F., Desbruslais, M., Joiner, S., Sidle, K. C. L., Gowland, I. and Collinge, J. (1997): The same
prion strain causes vCJD and BSE. Nature, 389, 448-450. Matravers, W., Bridgeman, J. and Smith, M.-F. (ed.)(2000): The BSE
Inquiry. p. 37. vol. 16. The Stationery Office Ltd., Norwich, UK. Casalone, C., Zanusso, G., Acutis, P. L., Crescio, M. I., Corona,
C., Ferrari, S., Capobianco, R., Tagliavini, F., Monaco, S. and Caramelli, M. (2003): Identification of a novel molecular and
neuropathological BSE phenotype in Italy. International Conference on Prion Disease: from basic research to intervention
concepts. Gasreig, Munhen, October 8-10. Bicaba, A. G., Laplanche, J. L., Ryder, S. and Baron, T. (2003): A molecular variant of
bovine spongiform encephalopatie. International Conference on Prion Disease: from basic research to intervention concepts.
Gasreig, Munhen, October 8-10. Asante, E. A., Linehan, J. M., Desbruslais, M., Joiner, S., Gowland, I., Wood, A. L., Welch, J.,
Hill, A. F., Lloyd, S. E., Wadsworth, J. D. F. and Collinge, J. (2002). BSE prions propagate as either variant CJD-like or sporadic
CJD-like prion strains in transgenic mice expressing human prion protein. EMBO J., 21, 6358-6366.
9/13/2005
Page 12 of 17
SEE SLIDES IN PDF FILE;
http://www.nih.go.jp/JJID/56/221.pdf
http://www.fsis.usda.gov/OPPDE/Comments/03-025IFA/03-025IFA-2.pdf
http://www.fsis.usda.gov/OPPDE/Comments/2006-0011/2006-0011-1.pdf
Subject: OPINION: BATCH testing of TSE rapid tests: SAMPLE SELECTION AND TEST SENSITIVITY ISSUES
Date: March 24, 2007 at 8:23 am PST
Opinion of the Scientific Panel BIOHAZ on the CRL report on batch testing of TSE rapid tests: sample selection and test sensitivity issues
Adopted on 7 March 2007. (Question Nº EFSA-Q-2006-204)
Last updated: 23 March 2007 Publication Date: 23 March 2007
Summary
The European Food Safety Authority (EFSA) and its Scientific Panel on Biological Hazards and the Expert Working Group on Transmissible Spongiform Encephalopathy (TSE) Testing were asked by the European Commission (EC) to evaluate a report of the Community Reference Laboratory (CRL) on batch testing of TSE rapid test kits which highlighted some matters of concern including sample selection and test sensitivity issues. At present, 12 rapid BSE test kits are approved by the EC for the post mortem testing of slaughtered cattle in accordance with the TSE Regulation (EC) No 999/2001.
The aim of a “Batch testing” programme is to compare different batches of a particular test kit for consistency of performance. A panel of samples is tested using each new batch of kits produced. The results obtained must fall within pre-determined limits. Batch release testing and /or approval are carried out to varying degrees by Member States. In order to establish a European wide batch testing procedure the CRL has assembled a panel of brain homogenates prepared from BSE positive bovine brain to be used for batch testing purposes. This sample panel was tested by the test manufacturers in their own laboratories using EU approved rapid tests. Most of the tests identified all of the positive samples in the set as positive, with medium to high readings. However, several of the tests failed to detect some of the positive samples, including some strongly positive samples. The CRL prepared a report on the testing and this was communicated to the companies concerned. These companies were given time to respond to the report and their replies were forwarded together with the CRL report to the EFSA for evaluation.
The experts of the Scientific Panel on Biological Hazards (BIOHAZ Panel) reviewed the CRL report on batch testing data and concluded that not all of the nine tests evaluated performed equally. The implications of this are twofold; firstly, the sample panel cannot be used in its current state to provide a batch testing system for all currently approved EU BSE rapid tests, although it is suitable for most of them. Secondly, they also suggest that there are profound differences in performance in terms of robustness, with respect to sample format, displayed by currently approved rapid tests. Consequentially, any observed differences in performance, if real, would be of concern. The observation that aliquots of the same positive sample were found to be highly positive according to some of the approved rapid tests but negative according to others, could be attributable to aspects of the test performance and/or to properties of the sample material tested. These concerns are addressed in a number of recommendations, as formulated in the Opinion.
The BIOHAZ Panel further concluded that these batch testing data do not compromise the previous Institute for Reference Materials and Measurements (IRMM)-EFSA evaluation of rapid BSE tests.
________________________________
1 For citation purposes: Opinion of the Scientific Panel on Biological Hazards on a request from the European Commission on the CRL report on batch testing of TSE rapid tests: sample selection and test sensitivity issues, The EFSA Journal (2007), 443, 1-18.
http://www.efsa.europa.eu/en/science/biohaz/biohaz_opinions/ej445_batch.html
Opinion
http://www.efsa.europa.eu/etc/medialib/efsa/science/biohaz/biohaz_opinions/ej445_batch.Par.0001.File.dat/biohaz_op_ej445_crl_batch_en.pdf
Summary
http://www.efsa.europa.eu/etc/medialib/efsa/science/biohaz/biohaz_opinions/ej445_batch.Par.0002.File.dat/biohaz_op_ej445_crl_batch_summary_en.pdf
TSS