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From: TSS ()
Subject: Re: Canada’s Protocols for BSE Surveillance DOES NOT MANDATE WB CONFIRMATION
Date: August 6, 2005 at 10:05 am PST

In Reply to: Canada’s Protocols for BSE Surveillance DOES NOT MANDATE WB CONFIRMATION posted by TSS on August 4, 2005 at 1:16 pm:

##################### Bovine Spongiform Encephalopathy #####################

Hello Karin,

indeed i did read the last line, i just think any confusion should be
eliminated. i think with the word 'or' is just asking for trouble. one
persons opinion on a 'poor quality' tissue sample, may not be the same as
the next persons opinion. so why even have the potential for confusion
there? i am sure you are aware of this, but maybe some other newer members
on the list have not read this study;

Effect of Tissue Deterioration on Postmortem BSE Diagnosis by
Immunobiochemical Detection of an Abnormal Isoform of Prion Protein
Hiroko HAYASHI1), Masuhiro TAKATA1), Yoshifumi IWAMARU1), Yuko USHIKI1)2),
Kumiko M. KIMURA1), Yuichi TAGAWA1), Morikazu SHINAGAWA1) and Takashi
1) Prion Disease Research Center, National Institute of Animal Health
2) Nippi Research Institute of Biomatrix
(Received 25-Aug-2003)
(Accepted 14-Jan-2004)
ABSTRACT. Surveillance for bovine spongiform encephalopathy (BSE) in fallen
stock in Japan is conducted with a commercial enzyme-linked immunosorbent
assay (ELISA) for mass screening, with Western blotting (WB) and
immunohistochemistry performed for confirmation of the ELISA. All tests are
based on immunological detection of an abnormal isoform of the prion protein
(PrPSc) in brain tissues, which have sometimes deteriorated by the time
samples from fallen stock reach a diagnostic laboratory. To evaluate BSE
surveillance procedures for fallen stock, we examined PrPSc detection from
artificially deteriorated BSE-affected bovine brain tissues with a
commercial ELISA kit and compared the results with those of WB. The optical
density (OD) values of the ELISA decreased with advancing deterioration of
the tissues, whereas no reduction in the signal for PrPSc was observed in
WB, even when performed after 4 days of incubation at 37°C. The progressive
decrease in the OD values in the ELISA appear to be caused by a partial loss
of the N-terminal moiety of PrPSc due to digestion by endogeneous and/or
contaminated microbial enzymes, and by the presence of ELISA inhibitors that
are generated in deteriorated tissues. These results suggest that WB is the
most reliable test for fallen stock, especially for cattle brains within
decaying carcasses.
KEY WORDS: BSE, ELISA, PrPSc, Western blot

[PDF (96K)] [References]

To cite this article:
Hiroko HAYASHI, Masuhiro TAKATA, Yoshifumi IWAMARU, Yuko USHIKI, Kumiko M.
KIMURA, Yuichi TAGAWA, Morikazu SHINAGAWA and Takashi YOKOYAMA “Effect of
Tissue Deterioration on Postmortem BSE Diagnosis by Immunobiochemical
Detection of an Abnormal Isoform of Prion Protein”. J. Vet. Med. Sci.. Vol.
66: 515-520. (2004) .


Another problem in testing fallen stock for BSE may arise from unequal
distribution of PrPSc in BSE-affected brains. Spongiform changes and
accumulation of PrPSc are most frequently observed in the obex region [15,
18], but, it could be quite difficult to collect the obex region precisely
from extensively deteriorated and liquefied brain tissue. Furthermore, in
such cases it would be difficult to perform IHC as a confirmation test.

It has been shown that sample autolysis does not affect detection of PrPSc
by means of WB [3, 5, 13]. Our WB results also demonstrated no reduction in
the PrPSc signal as a result of deterioration at 30*C or 37*C for up to 4
days, as so far examined (Figs. 2A and 3A). In this study, we showed that
several problems undermine the utility of the ELISA with deteriorated
samples, whereas WB remains very dependable. Therefore, WB might be the only
reliable procedure to detect PrPSc in severely damaged samples from fallen


Karin writes;

> I would be more worried about the latest USA suspect where no WB can be
done, due to formalin fixation of the sample. I don’t know if the
“reference” laboratory in Weybridge has ever missed any BSE-positive cattle
(or atypical bovine TSEs), but they have certainly failed to confirm several
cases of atypical scrapie, because they insisted on using the so-called
validated methods recommended by the OIE before 2003. I hope they now have
solved this problem.

i agree with this. in fact, the OIE since adhering to GWs BSE MRR policy
(minimal risk region), and doing away with the USA, Canada, and Mexico BSE
GBR Risk assessments, was anything but 'sound science'. by the OIE adhering
to this 'junk science' of this administrations corporate scientists, the OIE
has done nothing more than become a commodity brokerage for the legal
trading of all phenotypes of TSEs Globally. THEY have in essence done away
with 30 years of trying to eradicate TSEs globally. MILLIONS and millions of
dollars down the drain, with MILLIONS and millions of humans and animals now
becoming exposed even more than before, due to nothing more than greed and
the almighty buck. as GW says, ''bring em on''. he will get exactly what he
asks for again in the years and decades to come. but, as my birthday card
today states;

This birthday you have something to be thankful for (with a picture of GW on
the front),
I CAN'T RUN AGAIN....................amen!

SO why should he care, he will not be in office, but the markets will be
o.k. for now, borders open, and the list of demented and dead will continue
to slowly grow even more, with more strains of TSE mutating and being
exported throughout the globe...gee thanks GW/OIE!


Working Group Report on the Assessment of the Geographical BSE-Risk (GBR
III) of USA 2004 ''extremely/very unstable BSE/cattle system''




Canada and the United States have been raised to level III (presence of BSE
likely but not confirmed, or confirmed at a lower level) following a new
assessment taking into account the most recent evidence. EFSAs Scientific
Expert Working Group on geographic BSE risk assessment also evaluated the
status of Mexico and South Africa which were classified as level III.

From: Terry S. Singeltary Sr. []
Sent: Tuesday, July 29, 2003 1:03 PM
Cc:;; BSE-L
Subject: Docket No. 2003N-0312 Animal Feed Safety System [TSS SUBMISSION
TO DOCKET 2003N-0312]

Greetings FDA,


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
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

kindest regards,

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