WAHIS, WOAH, OIE, REPORT Switzerland Bovine Spongiform Encephalopathy Atypical L-Type
Switzerland Bovine Spongiform Encephalopathy Atypical L-Type
Switzerland - Bovine spongiform encephalopathy - Immediate notification
GENERAL INFORMATION
COUNTRY/TERRITORY OR ZONE
COUNTRY/TERRITORY
ANIMAL TYPE
TERRESTRIAL
DISEASE CATEGORY
OIE-listed
EVENT ID
4962
DISEASE
Bovine spongiform encephalopathy
CAUSAL AGENT
Bovine spongiform encephalopathy prion, atypical strain, L-type
GENOTYPE / SEROTYPE / SUBTYPE
-
START DATE
2023/03/03
REASON FOR NOTIFICATION
Recurrence of an eradicated disease
DATE OF LAST OCCURRENCE
2020/02/03
CONFIRMATION DATE
2023/03/08
EVENT STATUS
On-going
END DATE
-
SELF-DECLARATION
NO
REPORT INFORMATION
REPORT NUMBER
Immediate notification
REPORT ID
IN_159813
REPORT REFERENCE
BSE_2023_GR
REPORT DATE
2023/03/13
REPORT STATUS
Validated
NO EVOLUTION REPORT
-
EPIDEMIOLOGY
SOURCE OF EVENT OR ORIGIN OF INFECTION
Unknown or inconclusive
EPIDEMIOLOGICAL COMMENTS
The 12-year-old cow from a holding in the Canton of Grisons has been sampled at slaughter in the framework of the BSE surveillance program on 27 February 2023. The brain stem material was tested positive by IDEXX HerdChek BSE-Scrapie Ag Test on 03 March 2023. The result was confirmed on 08 March 2023 by Western lmmunoblot at the National Reference Laboratory for Transmissible Spongiform Encephalopathies (Neurocenter, Vetsuisse Faculty, Berne, Switzerland). The pathological prion protein was classified as (L) -Type, therefore the case has been categorized as atypical BSE. The entire carcass was disposed of as category 1 material.
QUANTITATIVE DATA SUMMARY
MEASURING UNIT
Animal
SpeciesSusceptibleCasesDeathsKilled and Disposed ofSlaughtered/ Killed for commercial useVaccinated
Cattle (DOMESTIC)NEW1910010TOTAL1910010
DIAGNOSTIC DETAILS
CLINICAL SIGNS
NO
METHOD OF DIAGNOSTIC
Diagnostic test
Test name Laboratory Species sampled Outbreaks Result date Result
Antigen detection Western blot (Ag Western blot) NeuroCenter, Department of Clinical Research and Veterinary Public Health (DCR-VPH), Division of Experimental Clinical Research, University of Bern Cattle Prättigau-Davos 2023/03/08 Positive
CONTROL MEASURES AT EVENT LEVEL
CONTROL MEASURES AT EVENT LEVEL
DOMESTIC ANIMALS
WILD ANIMALS
Traceability
Applied
Selective killing and disposal
Applied
Official disposal of carcasses, by-products and waste
Applied
Disinfection
Applied
Official destruction of animal products
Applied
Ante and post-mortem inspections
To be applied
OIE Conclusions on transmissibility of atypical BSE among cattle
Given that cattle have been successfully infected by the oral route, at least for L-BSE, it is reasonable to conclude that atypical BSE is potentially capable of being recycled in a cattle population if cattle are exposed to contaminated feed. In addition, based on reports of atypical BSE from several countries that have not had C-BSE, it appears likely that atypical BSE would arise as a spontaneous disease in any country, albeit at a very low incidence in old cattle. In the presence of livestock industry practices that would allow it to be recycled in the cattle feed chain, it is likely that some level of exposure and transmission may occur. As a result, since atypical BSE can be reasonably considered to pose a potential background level of risk for any country with cattle, the recycling of both classical and atypical strains in the cattle and broader ruminant populations should be avoided.
Annex 7 (contd) AHG on BSE risk assessment and surveillance/March 2019
34 Scientific Commission/September 2019
3. Atypical BSE
The Group discussed and endorsed with minor revisions an overview of relevant literature on the risk of atypical BSE being recycled in a cattle population and its zoonotic potential that had been prepared ahead of the meeting by one expert from the Group. This overview is provided as Appendix IV and its main conclusions are outlined below. With regard to the risk of recycling of atypical BSE, recently published research confirmed that the L-type BSE prion (a type of atypical BSE prion) may be orally transmitted to calves1 . In light of this evidence, and the likelihood that atypical BSE could arise as a spontaneous disease in any country, albeit at a very low incidence, the Group was of the opinion that it would be reasonable to conclude that atypical BSE is potentially capable of being recycled in a cattle population if cattle were to be exposed to contaminated feed. Therefore, the recycling of atypical strains in cattle and broader ruminant populations should be avoided.
The Group acknowledged the challenges in demonstrating the zoonotic transmission of atypical strains of BSE in natural exposure scenarios. Overall, the Group was of the opinion that, at this stage, it would be premature to reach a conclusion other than that atypical BSE poses a potential zoonotic risk that may be different between atypical strains.
4. Definitions of meat-and-bone meal (MBM) and greaves
snip...
REFERENCES
SNIP...END SEE FULL TEXT;
Consumption of L-BSE–contaminated feed may pose a risk for oral transmission of the disease agent to cattle.
Thus, it is imperative to maintain measures that prevent the entry of tissues from cattle possibly infected with the agent of L-BSE into the food chain.
''H-TYPE BSE AGENT IS TRANSMISSIBLE BY THE ORONASAL ROUTE''
This study demonstrates that the H-type BSE agent is transmissible by the oronasal route. These results reinforce the need for ongoing surveillance for classical and atypical BSE to minimize the risk of potentially infectious tissues entering the animal or human food chains.
Switzerland OIE Bovine spongiform encephalopathy atypical BSE type L TSE Prion
OIE Bovine spongiform encephalopathy, Switzerland
Information received on 05/02/2020 from Mr Hans Wyss, Chief Veterinary Officer, Schwarzenburgstrasse 161, Swiss Federal Veterinary Office, LIEBEFELD BERNE, Switzerland
Summary
Report type Immediate notification (Final report)
Date of start of the event 29/01/2020
Date of confirmation of the event 03/02/2020
Report date 05/02/2020
Date submitted to OIE 05/02/2020
Date event resolved 03/02/2020
Reason for notification Recurrence of a listed disease
Date of previous occurrence 03/2012
Manifestation of disease Clinical disease
Causal agent Prion (atypical BSE type L)
Nature of diagnosis Laboratory (advanced)
This event pertains to the whole country
New outbreaks (1)
Outbreak 1 Einsiedeln, Schwyz
Date of start of the outbreak 29/01/2020
Outbreak status Resolved (03/02/2020)
Epidemiological unit Farm
Affected animals
Species Susceptible Cases Deaths Killed and disposed of Slaughtered
Cattle 68 1 0 0 1
Summary of outbreaks Total outbreaks: 1
Total animals affected
Species Susceptible Cases Deaths Killed and disposed of Slaughtered
Cattle 68 1 0 0 1
Outbreak statistics
Species Apparent morbidity rate Apparent mortality rate Apparent case fatality rate Proportion susceptible animals lost*
Cattle 1.47% 0.00% 0.00% 1.47%
*Removed from the susceptible population through death, destruction and/or slaughter
Epidemiology
Source of the outbreak(s) or origin of infection
Unknown or inconclusive
Epidemiological comments
Confirmation of atypical Bovine spongiform encephalopathy (BSE) in a cow on 3 February 2020. The 13 year old cow from a holding in the Canton of Schwyz was subject to emergency-slaughter on 23 January 2020. Samples of the brain stem were sent in for analysis and confirmed at the Swiss National Reference Laboratory for Transmissible Spongiform Encephalopathies. The sample tested positive in the rapid test for BSE "Priostrip BSE" and was confirmed by Western lmmunoblot. The pathological prion protein could be classified as (L) -Type BSE and therefore as an atypical case of BSE. The entire carcass was disposed of as category 1 material.
Control measures
Measures applied
Vaccination prohibited
No treatment of affected animals
Measures to be applied
No other measures
Diagnostic test results
Laboratory name and type Species Test Test date Result
National Reference Laboratory (National laboratory) Cattle western blot 03/02/2020 Positive
Future Reporting
The event is resolved. No more reports will be submitted.
Map of outbreak locations
A descriptive study of the prevalence of atypical and classical scrapie in sheep in 20 European countries
Prevalence estimates of AS were higher in the fallen stock stream than in the healthy slaughter stream in four countries (Finland, Norway, Portugal and Switzerland).
Published: 18 April 2010
Surveillance and simulation of bovine spongiform encephalopathy and scrapie in small ruminants in Switzerland
Chantal Häusermann, Heinzpeter Schwermer, Anna Oevermann, Alice Nentwig, Andreas Zurbriggen, Dagmar Heim & Torsten Seuberlich BMC Veterinary Research volume 6, Article number: 20 (2010)
Abstract
Background
After bovine spongiform encephalopathy (BSE) emerged in European cattle livestock in 1986 a fundamental question was whether the agent established also in the small ruminants' population. In Switzerland transmissible spongiform encephalopathies (TSEs) in small ruminants have been monitored since 1990. While in the most recent TSE cases a BSE infection could be excluded, for historical cases techniques to discriminate scrapie from BSE had not been available at the time of diagnosis and thus their status remained unclear. We herein applied state-of-the-art techniques to retrospectively classify these animals and to re-analyze the affected flocks for secondary cases. These results were the basis for models, simulating the course of TSEs over a period of 70 years. The aim was to come to a statistically based overall assessment of the TSE situation in the domestic small ruminant population in Switzerland.
Results
In sum 16 TSE cases were identified in small ruminants in Switzerland since 1981, of which eight were atypical and six were classical scrapie. In two animals retrospective analysis did not allow any further classification due to the lack of appropriate tissue samples. We found no evidence for an infection with the BSE agent in the cases under investigation. In none of the affected flocks, secondary cases were identified. A Bayesian prevalence calculation resulted in most likely estimates of one case of BSE, five cases of classical scrapie and 21 cases of atypical scrapie per 100'000 small ruminants. According to our models none of the TSEs is considered to cause a broader epidemic in Switzerland. In a closed population, they are rather expected to fade out in the next decades or, in case of a sporadic origin, may remain at a very low level.
Conclusions
In summary, these data indicate that despite a significant epidemic of BSE in cattle, there is no evidence that BSE established in the small ruminant population in Switzerland. Classical and atypical scrapie both occur at a very low level and are not expected to escalate into an epidemic. In this situation the extent of TSE surveillance in small ruminants requires reevaluation based on cost-benefit analysis.
FRIDAY, MARCH 16, 2012
atypical BSE Mad cow disease found in Switzerland March 12, 2012
Subject: atypical BSE Mad cow disease found in Switzerland
March 12, 2012
From: "Terry S. Singeltary Sr." <flounder9@VERIZON.NET>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@LISTS.AEGEE.ORG>
Date: Fri, 16 Mar 2012 12:48:54 -0500
Monday, 12 March, 2012, 13:52
Mad cow strain found in Switzerland The Federal Veterinary Office has confirmed one case of mad cow disease in the canton of Bern. The cow was slaughtered last month. The case was discovered as part of a monitoring programme implemented by Swiss authorities. According to vets the cow was not infected with a classical case of BSE, but rather an atypical strain. They say that means the disease was not triggered by a certain animal feed outlawed in 2005. The cow was imported to Switzerland in 2006. Until this case, no cows with BSE have been found in Switzerland since 2007. The Federal Veterinary Office confirms it may have found another one of these atypical cases but stresses this strain of disease is rare. BSE was first diagnosed in 1990, and since then 467 cows were diagnosed with it in Switzerland. No cases of the human strain have ever been detected here.
on the official government site for BSE, it’s not in English. ...TSS
BSE-Fall bei einer importierten Kuh Bern, 12.03.2012 - Im Rahmen des offiziellen Überwachungsprogrammes wurde bei einer Kuh eines Betriebes im Kanton Bern BSE festgestellt. Analysen zeigen jedoch, dass es sich nicht um einen Fall von klassischer BSE handelt, sondern um eine sogenannte atypische BSE. Das Tier stammte aus Deutschland und wurde 2006 in die Schweiz importiert. Der aktuelle Fall ändert nichts an der guten Lage bezüglich BSE in der Schweiz. Bis 2006 hatten sich die Fälle stark vermindert, zwischen 2007 und 2011 wurden überhaupt keine Fälle mehr diagnostiziert. Aber die letzten Fälle erinnern daran, dass wir trotz dieser vorteilhaften Situation doch vereinzelt mit Fällen von BSE rechnen müssen. Die Diagnostik des Referenzlabors zeigt, dass es sich nicht um einen typischen Fall von BSE, sondern um einen sogenannt atypischen BSE-Fall handelt. Seit einigen Jahren werden in verschiedenen europäischen Ländern vereinzelt solche atypische Fälle von BSE festgestellt. Die Wissenschaftler glauben, dass diese Form der BSE möglicherweise eine sporadisch auftauchende Krankheit ist, die nicht auf infektiöses Material im Futter zurückzuführen ist. Im Gegensatz dazu werden die typischen BSE-Fälle durch infektiöses Material im Futter der Tiere verursacht. Beim verseuchten Tier handelt es sich um eine bald 7-jährige Kuh welche im April 2005 in Deutschland geboren und im November 2006 in die Schweiz importiert wurde. Die Kuh musste wegen Festliegens nach der Geburt geschlachtet werden und wurde deshalb routinemässig labordiagnostisch untersucht. Dabei wurde BSE diagnostiziert. Seit dem ersten Schweizer Fall im Jahre 1990 wurden in der Schweiz insgesamt 467 BSE-Fälle gemeldet. Mit Ausnahme eines Falles in 2011 waren alle betroffenen Rinder vor dem totalen Tiermehlverfütterungsverbot vom Januar 2001 geboren worden. Seit 1990 war im Laufe der Jahre eine ganze Reihe von Massnahmen beschlossen und umgesetzt worden, um die Krankheit bei den Rindern einzudämmen. Es wurde in der Schweiz nie ein Fall der Variante der Creutzfeldt-Jakob-Erkrankung bei Menschen, die mit BSE in Verbindung gebracht wird, diagnostiziert. Weitere Informationen finden sie auf der Web-Seite des BVET unter http://www.bvet.admin.ch >
BSE. Sie finden dort insbesondere die Geschichte der getroffenen Massnahmen sowie Grafiken zur Entwicklung der BSE-Fälle in der Schweiz. Adresse für Rückfragen: Regula Kennel, Bundesamt für Veterinärwesen Tel.: 031 323 84 96 Herausgeber: Bundesamt für VeterinärwesenInternet: http://www.bvet.admin.ch Volltextsuche Suche erweiterte Suche Die Bundesbehörden der Schweizerischen Eidgenossenschaft webmaster@admin.ch | Rechtliche Grundlagen
also, atypical bse and feed IS A FACTOR !
amplification and transmission of atypical BSE via feed is real, and can transmit just like the typical BSE did. any other statement is NOT based on science, but on TRADE $$$
P.9.21 Molecular characterization of BSE in Canada
Jianmin Yang1, Sandor Dudas2, Catherine Graham2, Markus Czub3, Tim McAllister1, Stefanie Czub1 1Agriculture and Agri-Food Canada Research Centre, Canada; 2National and OIE BSE Reference Laboratory, Canada; 3University of Calgary, Canada
Background: Three BSE types (classical and two atypical) have been identified on the basis of molecular characteristics of the misfolded protein associated with the disease. To date, each of these three types have been detected in Canadian cattle.
Objectives: This study was conducted to further characterize the 16 Canadian BSE cases based on the biochemical properties of there associated PrPres.
Methods: Immuno-reactivity, molecular weight, glycoform profiles and relative proteinase K sensitivity of the PrPres from each of the 16 confirmed Canadian BSE cases was determined using modified Western blot analysis.
Results: Fourteen of the 16 Canadian BSE cases were C type, 1 was H type and 1 was L type. The Canadian H and L-type BSE cases exhibited size shifts and changes in glycosylation similar to other atypical BSE cases. PK digestion under mild and stringent conditions revealed a reduced protease resistance of the atypical cases compared to the C-type cases. N terminal- specific antibodies bound to PrPres from H type but not from C or L type. The C-terminal-specific antibodies resulted in a shift in the glycoform profile and detected a fourth band in the Canadian H-type BSE.
Discussion: The C, L and H type BSE cases in Canada exhibit molecular characteristics similar to those described for classical and atypical BSE cases from Europe and Japan. This supports the theory that the importation of BSE contaminated feedstuff is the source of C-type BSE in Canada. It also suggests a similar cause or source for atypical BSE in these countries.
*** It also suggests a similar cause or source for atypical BSE in these countries.
snip...end...BSE Listserve Archives...tss
FRIDAY, MARCH 16, 2012
atypical BSE Mad cow disease found in Switzerland March 12, 2012
Saturday, November 19, 2011
Novel Prion Protein in BSE-affected Cattle, Switzerland
Subject: BSE almost eradicated in Switzerland
From: "Terry S. Singeltary Sr." <flounder9@VERIZON.NET>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@LISTS.AEGEE.ORG>
Date: Tue, 20 Dec 2005 09:42:50 -0600 Content-Type: text/plain Parts/Attachments: text/plain (62 lines)
##################### Bovine Spongiform Encephalopathy #####################
BSE almost eradicated in Switzerland
For the second year in a row just three cases of BSE, or mad cow disease, were diagnosed in Switzerland in 2005. The disease appears to be close to eradication. Switzerland has been cited as a good role model in the fight against Bovine Spongiform Encephalopathy and Swiss expertise has been deployed abroad.
Related links: » UN Food and Agriculture Organization » Federal Veterinary Office » World Organisation for Animal Health (BSE cases worldwide)
Just three new cases of BSE were detected in Switzerland in 2005
The latest figures confirm the steady fall in the numbers of new BSE cases in the national herd since 2001.
Marcel Falk of the Federal Veterinary Office told swissinfo that it was now possible to "see the beginning of the end of BSE in Switzerland".
He said the fact that the number of cases had remained stable in 2005 confirmed that the drop seen in 2004 - from 21 to just three – was more than merely a statistical blip.
Falk said it was impossible to say when BSE might finally disappear from Switzerland, given that the incubation period for the disease was at least five years, and could be as long as ten to 12 years.
But he said the three cows diagnosed with the disease this year were likely to have contracted it several years ago.
"One knows from experiments that all cows [which contract the disease] are infected in the first year of life. The cows diagnosed in 2005 were all between seven and ten years old, which means they contracted the disease in the mid-1990s."
Controlling the disease
In 1990 Switzerland became the third European country after Britain and Ireland to register cases of BSE in its cattle. BSE was first defined in Britain in November 1986. Some 83,000 cases have been detected there since then.
The human disease, Variant Creutzfeldt-Jakob Disease (vCJD), was recognised in 1996 and is thought to result from the consumption of BSE-infected meat. There have been no cases of vCJD reported in Switzerland.
Last year, the United Nations praised Switzerland for its efforts to control mad cow disease, calling it a model for other nations.
Various measures aimed at controlling the disease have been introduced in Switzerland, including a complete ban on animal products in feed for livestock in 2001.
Andrew Speedy of the UN's Food and Agriculture Organization said testing was begun as soon as BSE was found in Swiss cattle.
"The control measures which we recommended – the ban on meat and bonemeal and the destruction of specified risk materials – were done right from the word go," Speedy said.
In 2004, Switzerland sent BSE experts to the United States, following that country's first confirmed case of BSE.
swissinfo with agencies
(swissinfo, 20.12.2005)
TSS
#################### https://lists.aegee.org/bse-l.html ####################
Subject: Switzerland reports 2 new BSE cases
From: "Terry S. Singeltary Sr." <flounder@WT.NET>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE>
Date: Tue, 16 Sep 2003 11:54:11 -0500
Tuesday 16.09.2003, CET 18:50
September 16, 2003 4:01 PM
New BSE cases
Two new mad cow disease, or BSE, cases have been reported in Switzerland since August this year.
The Federal Veterinary Office said both infected animals were born after restricitons on animal feed were tightened in 1996.
Authorities have recorded a total of 16 BSE cases in Switzerland this year, in comparison to 24 in 2002.
TSS
Subject: SWITZERLAND: Reports Three More Cases BSE
From: "Terry S. Singeltary Sr." <flounder@WT.NET>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE>
Date: Wed, 4 Apr 2001 14:04:49 -0700 Content-Type: text/plain Parts/Attachments: text/plain (41 lines)
######### Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE> #########
Switzerland: Reports Three More Cases BSE
Apr 04, 2001 (FWN Financial via COMTEX) -- Hamburg, April 4 (BridgeNews) - Three more cases of Bovine Spongiform Encephalopathy (BSE), or mad cow disease have been found in Switzerland, the Swiss Federal Veterinary Agency said Wednesday.
This brings the number of confirmed cases to eight in 2001.
One case was in the Zug canton (local government area), one in the Bern canton and one in Schwyz.
In 2000, Switzerland had 33 BSE cases.
* * *
Switzerland imposed a ban on feeding meat and bone meal (MBM) to cattle in May 1996.
Greetings List Members,
amazing what you can find when you are looking. amazing what you don't find, when you are not looking. a fine example of the later would be in the United States. I suppose the U.S. could be BSE free forever if we never look $$$
yes, the U.S. can debate which test they will use 'for ten years' and the whole time, the U.S. will be BSE free. the 950 SCRAPIE INFECTED FLOCKS will not matter...
(NOT)
thank you, Terry S. Singeltary Sr., Bacliff, Texas USA
############ http://mailhost.rz.uni-karlsruhe.de/warc/bse-l.html ############
Subject: BSE Switzerland...2000
From: "Terry S. Singeltary Sr." <flounder@wt.net>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE>
Date: Thu, 27 Apr 2000 15:00:59 -0700 Content-Type: text/plain Parts/Attachments: text/plain (31 lines)
######### Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE> #########
greetings everyone,
i was looking over the Swiss data, it would seem that they could very well double their BSE cases for the year 2000. with other E.U. Countries having increase in BSE, and the U.K. still putting out the biggest number, things do not look good. BSE will most likely turn into a world problem. the sooner the world realizes this, the sooner we can hopefully eradicate it. but that may prove to be very difficult, if every Country does not have the same guidelines, and those guidelines are enforced equally...
kind regards, Terry S. Singeltary Sr., Bacliff, Texas USA
--------------------------------------------
SWITZERLAND
Figures between () are cases detected under strict BSE-monitoring program
1998 1999 2000
14 25(25) 13(7)
as of April 25, 2000
---------------------------
http://www.admin.ch/bvet/tiergesundheit/d/ausbild_beratung/tierseuchen/ bse/bse-ch/kantone.html
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Subject: BSE Switzerland
From: "Terry S. Singeltary Sr." <flounder@wt.net>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE>
Date: Tue, 9 Nov 1999 08:24:42 -0600 Content-Type: text/plain Parts/Attachments: text/plain (21 lines)
Terry S. Singeltary Sr., Bacliff, Texas USA;
Greetings, it seems the new testing in Switzerland is working better than thought. I am beginning to see the big picture, as to why the U.S. refuses to use these tests...
ProMED source via Xinhua 5 Nov. 1999 (edited) --
Five new cases of mad cow disease (Bovine Spongiform Encephalopathy, BSE) have been discovered in Switzerland since mid-October 1999, bringing the total number of such cases to 41 this year, according to the Federal Veterinary Office Friday.
The office said 4 of the cows were born after the ban on contaminated animal feed was introduced in 1990.
Earlier this year, the government introduced rigorous testing of the cattle.
In the past 10 years, a total of 323 cases were found in Switzerland...
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Subject: Re: BSE Switzerland
From: Vincent Dedet <>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE>
Date: Tue, 9 Nov 1999 16:05:53 +0100
flounder@wt.net writes: >ProMED source via Xinhua 5 Nov. 1999 (edited) --
>Five new cases of mad cow disease (Bovine Spongiform Encephalopathy,
>BSE) have been discovered in Switzerland since mid-October 1999,
>bringing the total number of such cases to 41 this year, according to
>the Federal Veterinary Office Friday.
> >The office said 4 of the cows were born after the ban on contaminated >animal feed was introduced in 1990.
> >Earlier this year, the government introduced rigorous testing of the >cattle.
> >In the past 10 years, a total of 323 cases were found in Switzerland...
Hello to the list, I checked the Swiss website and saw that within these 5 cases, one seems to be non BAB, and among the 4 others : 1 with an age "under investigation" and 1 was born on the 11th of december 1995. This is, to my knowledge, the most recent date of birth for a BSE case ever (but I am not 100 % sure UK has not done better yet). We are coming closer and closer to the date of march 1996. When will we see a BAB case born after this date ? That will be puzzling everyone, because all the measures implemented after this date are supposed to have completely cut the transmission ways for BSE… Are consumers and crisis managers ready for when it comes up ? I wonder…
Vincent Dedet Auzalide Sante Animale 5, rue Charles Fourier 75013 PAris (France)
Tel : +33 1 45 65 96 06 Fax : +33 1 45 65 96 08 e-mail :
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Subject: Re: BSE Switzerland From: Roland Heynkes <>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE>
Date: Wed, 10 Nov 1999 09:02:14 +0100
Content-Type: text/plain Parts/Attachments: text/plain (76 lines)
Robert A. LaBudde answered Terry S. Singeltary Sr.:
>>Greetings, it seems the new testing in Switzerland is working better
>>than thought. I am beginning to see the big picture, as to why the U.S.
>>refuses to use these tests...
> > 1. The USDA does not 'refuse' to use these tests. In fact, it carries out > prion testing all referral samples as well as histopathological examination. > Terry wrote that the USDA refuses to use the Prionics test and as far as I know this is correct. The Prionics colleagues may correct me. But the USDA uses its own tests and at least in the past this has been much more expensive and very slow, but not necessarily less sensitive. It is ok when they use an evaluated immunological test, whereas a histopathological examination is not enough to exclude BSE.
> 2. USDA 'apparently' is not 'choosing' to run these tests on asymptomatic > animals. I say 'apparently', since I have seen no reports to the contrary. > This does not, however, mean they do not, only that we do not know for sure > that they do. Like you, I would be happier seeing a Swiss-style benchmark > evaluation of a random subset of, say, 10000 cattle prion-tested to confirm > the absence of ukBSE in the US herd. > The Prionics Test would identify not only ukBSE, but most likely also usBSE, because it marks scrapie and CJD too.
> 4. To be fair, the uniformly negative histopathological and prion-testing > results on referral samples present compelling evidence that testing > asymptomatic animals would be unproductive. If you can't find disease or > agent in referral samples where the incidence should be 100,000 times more > likely, why expect any positives in asymptomatic animals? Their 'apparent' > position is scientifically defensible. > Histopathological examination isn't able to exclude BSE, especially because we do not know if usBSE would show spongiform lesions and astrocytosis. I would be very interested in a reference for this "100,000 times more likely".
> 5. The issue also crucially hinges on the incidence of false positives in > the prion testing. Any test has a false positive rate for a large variety > of reasons, including faulty manufacture, cross-contamination in the lab, > mixed-up samples, aliasing fragments, etc. Only a person without experience > in such matters would believe otherwise. This false positive rate in the > best of circumstances is about 1:1,000-10,000 practically. So in a test of > a 10,000 animal cohort, all expected to show negative results, we might > very well find 1 or 2 false positives. Consider the negative implications > of such an event! Again, it is hard to fault the USDA for not taking on > such a large political risk of controversy for no scientific gain. > With the Prionics test we would not find 1 or 2 false positives in a country without BSE. It has internal controls that make this impossible. Furthermore each positive result of this test of course would lead to a reaxamination with other methods. Therefore false positive BSE test results are absolutely no problem.
> 6. A more interesting question is why the UK or Portugal or Belgium or > France do not carry out such studies on their own herds. Given the known > positives already being found in referral animals, they could expect a > measurable incidence (well beyond the false positive rate) in the > asymptomatic animals. Why not find out what that rate actually is? > This is indeed a good idea, but of course they are very afraid to find to many hidden cases. Don't forget that the identification of BSE infected cows is dangerous for business, whereas eating BSE cows is only dangerous for the consumers.
best regards
Roland
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Subject: 2 BSE cases in Switzerland
From: Marc Barbier <>
Reply To: Bovine Spongiform Encephalopathy <BSE-L@UNI-KARLSRUHE.DE>
Date: Mon, 1 Jan 1990 09:40:01 +0100
From Official sources (BVET)
Two new cases of "mad cow" in Switzerland
Two new cases of mad-cow have been confirmed in Switzerland (total of 291 till now).
These two cows were born in 1994 in Buren district (Berne Canton) and in Lucerne District 4 years after the MBM ban in 1990.
---------------
Are we facing the "Swiss efficiency" in systematic testing programme, in epidemiosurveillance and in veterinary control that allow to detect more cases than in other continental countries or is there something specific to the Swiss situation (a Swiss strain of agent) ? Perhaps some Swiss comments about it on this list ?
Marc BARBIER INRA ESR Domaine Universitaire BP 47X 38040 Grenoble cedex France TEL: 00.33.4.76.82.78.82
SNIP...END...TSS
Helen Pearson
Nature (2002)
BSE-related brain disease could emerge in different form.
The number of people dying from sporadic Creutzfeldt-Jakob disease (CJD) has risen sharply in Switzerland. The finding is raising fears that 'mad cow disease' could have spread to humans in another form1.
Lancet. 2002 Jul 13;360(9327):139-41.
Incidence of Creutzfeldt-Jakob disease in Switzerland.
Glatzel M1, Rogivue C, Ghani A, Streffer JR, Amsler L, Aguzzi A.
Author information
Abstract
The incidence of Creutzfeldt-Jakob disease (CJD) in Switzerland increased two-fold in 2001, and figures from the first quarter of 2002 indicate that it continues to rise. Neither age at onset nor duration of disease were different from previous years. Genetic analysis of the 27 reported cases revealed only one disease-associated mutation in the prion gene. None of the recognised risk factors for acquired CJD were reported on the official notification forms. Glycotype profiling, histopathology, and immunohistochemistry indicate that none of the cases fulfilled the definition of variant CJD, which is thought to be caused by bovine prions. Several scenarios could account for the increase in CJD, including improved reporting, iatrogenic transmission, and transmission of a prion zoonosis.
PMID: 12126826 DOI: 10.1016/S0140-6736(02)09384-4
Heightened incidence of sporadic Creutzfeldt-Jakob disease is associated with a shift in clinicopathological profiles
Stoeck, K; Hess, K; Amsler, L; Eckert, T; Zimmermann, D R; Aguzzi, A; Glatzel, M (2008).
Heightened incidence of sporadic Creutzfeldt-Jakob disease is associated with a shift in clinicopathological profiles.
Journal of Neurology, 255(10):1464-1472.
Abstract
Incidences of human transmissible spongiform encephalopathies are monitored by national registries in the majority of countries in Western Europe. During the past 13 years incidences for Creutzfeldt-Jakob disease (CJD) in Switzerland fluctuated between 0.4 and 2.63 cases/10(6) inhabitants. We have compared clinicpathological patient profiles including geographic and gender distribution, age at disease onset, duration of disease, clinical symptoms, and recognized or hypothetical risk factors for CJD, genetic risk factors, biochemical and histopathological data for two cohorts of Swiss sporadic CJD patients from years of regular sporadic CJD incidence (1996-2000, mean incidence 1.3 cases/10(6) inhabitants, n = 47) to Swiss sporadic CJD patients from years of elevated sporadic CJD incidence (2001-2004, mean incidence 2.3 cases/10(6) inhabitants, n = 73). Sporadic CJD patients from the cohort with elevated sporadic CJD incidence presented with a higher frequency of rare sporadic CJD subtypes. Patients of these subtypes were significantly older and showed a skewed male/female ratio when compared to published patients of identical sporadic CJD-types or to patients from the 1996-2000 cohort and indicates that improved detection of rare sporadic CJD subtypes may have contributed to increased incidence.
Interesting note on recent BSE cases;
BRAZIL BSE START DATE 2023/01/18
BRAZIL BSE CONFIRMATION DATE 2023/02/22
BRAZIL BSE END DATE 2023/03/03
SPAIN BSE START DATE 2023/01/21
SPAIN BSE CONFIRMATION DATE 2023/02/03
SPAIN BSE END DATE 2023/02/06
NETHERLANDS BSE START DATE 2023/02/01
NETHERLANDS BSE CONFIRMATION DATE 2023/02/01
NETHERLANDS BSE END DATE 2023/03/13
2.3.2. New evidence on the zoonotic potential of atypical BSE and atypical scrapie prion strains
Olivier Andreoletti, INRA Research Director, Institut National de la Recherche Agronomique (INRA) – École Nationale Vétérinaire de Toulouse (ENVT), invited speaker, presented the results of two recently published scientific articles of interest, of which he is co-author:
‘Radical Change in Zoonotic Abilities of Atypical BSE Prion Strains as Evidenced by Crossing of Sheep Species Barrier in Transgenic Mice’ (MarinMoreno et al., 2020) and ‘The emergence of classical BSE from atypical/Nor98 scrapie’ (Huor et al., 2019).
In the first experimental study, H-type and L-type BSE were inoculated into transgenic mice expressing all three genotypes of the human PRNP at codon 129 and into adapted into ARQ and VRQ transgenic sheep mice. The results showed the alterations of the capacities to cross the human barrier species (mouse model) and emergence of sporadic CJD agents in Hu PrP expressing mice: type 2 sCJD in homozygous TgVal129 VRQ-passaged L-BSE, and type 1 sCJD in homozygous TgVal 129 and TgMet129 VRQ-passaged H-BSE.
This study demonstrates that the H-type BSE agent is transmissible by the oronasal route. These results reinforce the need for ongoing surveillance for classical and atypical BSE to minimize the risk of potentially infectious tissues entering the animal or human food chains.
***Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility.***
Even if the prevailing view is that sporadic CJD is due to the spontaneous formation of CJD prions, it remains possible that its apparent sporadic nature may, at least in part, result from our limited capacity to identify an environmental origin.
O.05: Transmission of prions to primates after extended silent incubation periods: Implications for BSE and scrapie risk assessment in human populations
Emmanuel Comoy, Jacqueline Mikol, Valerie Durand, Sophie Luccantoni, Evelyne Correia, Nathalie Lescoutra, Capucine Dehen, and Jean-Philippe Deslys Atomic Energy Commission; Fontenay-aux-Roses, France
Prion diseases (PD) are the unique neurodegenerative proteinopathies reputed to be transmissible under field conditions since decades. The transmission of Bovine Spongiform Encephalopathy (BSE) to humans evidenced that an animal PD might be zoonotic under appropriate conditions. Contrarily, in the absence of obvious (epidemiological or experimental) elements supporting a transmission or genetic predispositions, PD, like the other proteinopathies, are reputed to occur spontaneously (atpical animal prion strains, sporadic CJD summing 80% of human prion cases).
Non-human primate models provided the first evidences supporting the transmissibility of human prion strains and the zoonotic potential of BSE. Among them, cynomolgus macaques brought major information for BSE risk assessment for human health (Chen, 2014), according to their phylogenetic proximity to humans and extended lifetime. We used this model to assess the zoonotic potential of other animal PD from bovine, ovine and cervid origins even after very long silent incubation periods.
*** We recently observed the direct transmission of a natural classical scrapie isolate to macaque after a 10-year silent incubation period,
***with features similar to some reported for human cases of sporadic CJD, albeit requiring fourfold long incubation than BSE. Scrapie, as recently evoked in humanized mice (Cassard, 2014),
***is the third potentially zoonotic PD (with BSE and L-type BSE),
***thus questioning the origin of human sporadic cases.
We will present an updated panorama of our different transmission studies and discuss the implications of such extended incubation periods on risk assessment of animal PD for human health.
===============
***thus questioning the origin of human sporadic cases***
===============
***our findings suggest that possible transmission risk of H-type BSE to sheep and human. Bioassay will be required to determine whether the PMCA products are infectious to these animals.
==============
PRION 2015 CONFERENCE
***Transmission data also revealed that several scrapie prions propagate in HuPrP-Tg mice with efficiency comparable to that of cattle BSE. While the efficiency of transmission at primary passage was low, subsequent passages resulted in a highly virulent prion disease in both Met129 and Val129 mice.
***Transmission of the different scrapie isolates in these mice leads to the emergence of prion strain phenotypes that showed similar characteristics to those displayed by MM1 or VV2 sCJD prion.
***These results demonstrate that scrapie prions have a zoonotic potential and raise new questions about the possible link between animal and human prions.
PRION 2016 TOKYO
Saturday, April 23, 2016
SCRAPIE WS-01: Prion diseases in animals and zoonotic potential 2016
Prion. 10:S15-S21. 2016 ISSN: 1933-6896 printl 1933-690X online
Taylor & Francis
Prion 2016 Animal Prion Disease Workshop Abstracts
WS-01: Prion diseases in animals and zoonotic potential
Transmission of the different scrapie isolates in these mice leads to the emergence of prion strain phenotypes that showed similar characteristics to those displayed by MM1 or VV2 sCJD prion.
These results demonstrate that scrapie prions have a zoonotic potential and raise new questions about the possible link between animal and human prions.
http://www.tandfonline.com/doi/abs/10.1080/19336896.2016.1163048?journalCode=kprn20
Title: Transmission of scrapie prions to primate after an extended silent incubation period)
*** In complement to the recent demonstration that humanized mice are susceptible to scrapie, we report here the first observation of direct transmission of a natural classical scrapie isolate to a macaque after a 10-year incubation period. Neuropathologic examination revealed all of the features of a prion disease: spongiform change, neuronal loss, and accumulation of PrPres throughout the CNS.
*** This observation strengthens the questioning of the harmlessness of scrapie to humans, at a time when protective measures for human and animal health are being dismantled and reduced as c-BSE is considered controlled and being eradicated.
*** Our results underscore the importance of precautionary and protective measures and the necessity for long-term experimental transmission studies to assess the zoonotic potential of other animal prion strains.
***> UNITED STATES BSE TSE PRION OLD DAYS <***
Subject: U.S. Emergency Bovine Spongiform Encephalopathy Response Plan Summary
Date: Tue, 4 May 1999 18:25:12 -0500
From: "Terry S. Singeltary Sr."
Reply-To: Bovine Spongiform Encephalopathy
To: BSE-L@uni-karlsruhe.de
From: Terry S. Singeltary Sr., Bacliff, Texas......
I thought it might be interesting for those of you who have not seen this plan, to do so. So here it is...
The mission of the U.S. Department of Agriculture (USDA) is to enhance the quality of life for the American people by supporting production agriculture; ensuring a safe, affordable, nutritious, and accessible food supply; caring for agricultural, forest, and range lands; supporting sound development of rural communities; providing economic opportunities for farm and rural residents; expanding global markets for agricultural and forest products and services; and working to reduce hunger in America and throughout the world.
USDA's Animal and Plant Health Inspection Service (APHIS) is responsible for ensuring the health and care of animals and plants. APHIS improves agricultural productivity and competitiveness and contributes to the national economy and the public health. USDA's Food Safety and Inspection Service (FSIS) is responsible for protecting the Nation's meat and poultry supply--making sure it is safe, wholesome, unadulterated, and properly labeled and packaged. These two agencies have come together to lead USDA's actions in the prevention, monitoring, and control of bovine spongiform encephalopathy (BSE) in the U.S. livestock and food supply.
The public knows BSE as "MAD COW DISEASE", a disease linked to human cases of new-variant Creutzfeldt-Jakob disease (nvCJD). USDA knows BSE as the disease that devastated the livestock industry in the United Kingdom and shattered consumer confidence in Europe. BSE has affected international trade and all aspects of the animal and public health communities. It has called even greater attention to the U.S. Government's accountability for a safe food supply.
No case of BSE has ever been found in the United States. Since 1989, USDA has had a number of stringent safeguards in place to prevent BSE from entering the country. USDA conducts an ongoing, comprehensive interagency surveillance program for BSE. This surveillance program allows USDA to monitor actively for BSE to ensure immediate detection in the event that BSE were to be introduced into the United States. Immediate detection allows for swift response. As an emergency preparedness measure, USDA has developed this BSE Response Plan to be initiated in the event that a case of BSE is diagnosed in the United States. The Plan details comprehensive instructions for USDA staff as to who is to do what, when, where, and how in the event that BSE were to be diagnosed in the United States.
BACKGROUND
APHIS is responsible for being prepared for potential FOREIGN animal disease outbreaks. The purpose of such preparation is to provide a step-by-step plan of action in the event that a FOREIGN animal disease, such as BSE, is detected in the United States. These plans, often referred to as "RED BOOKS", provide guidance by outlining certain actions that should take place, such as identification of a suspect animal, laboratory confirmation, epidemiologic investigation, and animal and herd disposition activities. Copies of Red Books for specific FOREIGN animal diseases are distributed to agency headquarters and each regional and field office to have in preparation for a disease outbreak.
In 1990, APHIS developed a plan to respond to a confirmation of BSE in the United States. In August 1996, a joint APHIS-FSIS working group updated the BSE Red Book in accordance with current science and research surrounding BSE and the related family of disease called transmissible spongiform encephalopathies (TSE's). The BSE Red Book is officially entitled BSE EMERGENCY DISEASE GUIDELINES.
The APHIS-FSIS working group determined that the BSE Red Book, which detailed laboratory and field activities to be carried out in an emergency, needed another component. After the March 1996 announcement by the United Kingdom that BSE was linked to nvCJD, it became apparent to the working group that the Plan needed to address communication issues, both internally within USDA and the Federal Government and externally to the public at large. A confirmed case of BSE would affect such a vast array of stakeholders-consumers, cattle producers, the food animal industry, international trading partners, animal and public health communities, media, and others. Having clear, accurate information readily available would build trust and credibility and facilitate any response measures needed. There needed to be a notification plan. Who was responsible for notifying who, what, when, and how? The plan needed to identify clear channels of communication as to ensure immediate collection and dissemination of accurate information.
The joint APHIS--FSIS working group became formally known as the BSE Response Team and is responsible for the development of this BSE Response Response Plan. BSE Response Team members represent a mix of backgrounds and expertise, including veterinary medicine, food safety, public health, epidemiology, pathology, international trade, and public affairs. The Team is coordinated by two Team Leaders, one each from APHIS and FSIS, who serve as liaisons and technical advisors to their respective agencies on regulations and policies regarding BSE. Over the past 2 years, the BSE Response Plan has been reviewed, edited, revised, and approved by officials at all levels of APHIS, FSIS, and USDA. The Plan has also been shared with other Government agencies, such as the Food and Drug Administration (FDA), the Centers for Disease Control and Prevention (CDC), and the National Institutes of Health (NIH), and other stakeholders, such as the Animal Ag Coalition. The BSE Response Team monitors and assesses all ongoing events and research findings regarding TSE's. The Team leaders are responsible for ensuring that prevention and diagnostic measures are continually revised and adjusted as new information and knowledge become available.
NOTIFICATION: Roles and Responsibilities
Surveillance
As part of USDA's surveillance program for BSE in the United States, veterinary pathologists and field investigators from APHIS and FSIS have received training from British counterparts in diagnosing BSE. FSIS inspects cattle before they go to slaughter; these inspection procedures include identifying animals with central nervous system conditions. Animals with such conditions are considered suspect for BSE, prohibited from slaughter, and referred to APHIS for examination as explained below.
Pathologists at APHIS National Veterinary Services Laboratories (NVSL) histopathologically examine the brains from these condemned animals. In addition, samples are tested using a technique called immunohistochemistry, which tests for the presence of the protease-resistant prion protein (a marker for BSE). NVSL also examines samples from neurologically ill cattle and nonambulatory ("DOWNER") cattle identified on the farm or at slaughter and from rabies-negative cattle submitted to veterinary diagnostic laboratories and teaching hospitals.
NOTIFICATION
Because of their responsibility for examining condemned or BSE-suspect animals, NVSL is the organization responsible for activating the notification and BSE response process. It is NVSL that will begin the activation of the BSE Response Plan. From the time a sample is submitted, it takes 14 to 18 days to confirm a diagnosis of BSE In the first 10 to 13 days, pathologists at NVSL have enough information to either rule out BSE or determine the need for additional tests. If it is determined that there is no evidence of BSE, the results are added to the more than 7,500 others that have also been negative. NVSL maintains these data.
If additional tests do suggest a presumptive diagnosis of BSE, an NVSL pathologist will hand carry the sample to the United Kingdom for confirmation. It is at this critical point, when NVSL suggests a diagnosis of BSE and is preparing to send the sample to the United Kingdom, that this BSE Response Plan is initiated. The Plan begins the preliminary notification from NVSL to APHIS.
Preliminary Notification
The director of NVSL is responsible for immediately notifying the APHIS, Veterinary Services (VS) deputy administrator when tests suggest a presumptive diagnosis of BSE.
Once NVSL has made a presumptive diagnosis of BSE, APHIS and FSIS field activities will also be initiated. APHIS will receive notification (either confirming or not confirming NVSL's diagnosis) from the United Kingdom anywhere between 24 and 96 hours. (The international animal health community has recognized the United Kingdom's Central Veterinary Laboratory {CVL} as the world's reference laboratory for diagnosing BSE. Other countries, including Belgium, France, Ireland, Luxembourg, the Netherlands, Portugal, and Switzerland, have all sent samples to this lab to confirm their first case of BSE).
NVSL
NVSL will provide all laboratory support in carrying out this BSE Response Plan and serve as the liaison with CVL. NVSL will prepare its facility to receive and process additional samples from the suspect animal's progeny or herdmates or other suspects. NVSL will also coordinate any other assistance from State or university diagnostic laboratories if necessary.
APHIS, VS DEPUTY ADMINISTRATOR
Veterinary Services is the animal health arm of APHIS and the program responsible for carrying out field actions in response to BSE. Upon notification of a presumptive diagnosis from NVSL, the APHIS, VS deputy administrator immediately notifies the FSIS, Office of Public Health and Science (OPHS) deputy administrator. APHIS and FSIS deputy administrators will alert the BSE Response Team and activate the Response Plan. The VS deputy administrator serves as the liaison between the BSE Response Team and the APHIS administrator. The APHIS, VS deputy administrator notifies the APHIS administrator and the VS regional director of the State from which the suspect animal originated.
APHIS Administrator
The APHIS Administrator immediately notifies the USDA Assistant Secretary for Marketing and Regulatory Programs. This immediate notification will be followed by an official informational memorandum from the APHIS Administrator, through the Assistant Secretary for Marketing and Regulatory Programs, to the Secretary of Agriculture. This memorandum will be prepared by the BSE Response Team; a draft is maintained by the Team leaders in the reserved section of their plans. The APHIS Administrator is responsible for securing indemnity funds for depopulation of the herd if CVL confirms NVSL's diagnosis.
Assistant Secretary for Marketing and Regulatory Programs
The Assistant Secretary for Marketing and Regulatory Programs, in conjunction with the Undersecretary for Food Safety, is responsible for notifying the Secretary. The Assistant Secretary serves as the liaison between APHIS and Department-level officials.
Secretary of Agriculture
The Secretary has the authority to declare a Federal EMERGENCY if appropriate and approve funding as necessary. Information will be provided to the Secretary up the chain of command from the BSE Response Team.
FSIS, OPHS Deputy Administrator
The OPHS Deputy Administrator, together with the APHIS, VS Deputy Administrator, alert the BSE Response Team leaders and instruct them to assemble the BSE Response Team and activate the Plan. The OPHS Deputy Administrator serves as the liaison between the BSE Response Team and the FSIS Administrator. The OPHS Deputy Administrator is responsible for notifying the FSIS regional director in charge of the State from which the suspect animal originated.
FSIS Deputy Administrator
The FSIS Deputy Administrator is responsible for notifying the Undersecretary for Food Safety.
Undersecretary for Food Safety
The Undersecretary for Food Safety, in conjunction with the Assistant Secretary for Marketing and Regulatory Programs, notifies the Secretary of Agriculture.
APHIS, VS, Regional Director
The APHIS, VS regional director in charge of the State from which the suspect animal originated notifies the VS Area Veterinarian-in-Charge (AVIC) for that State. The regional director is the liaison between VS field staff and the VS Deputy Administrator at headquarters. In addition, the regional director shares all information with the BSE Response Team.
APHIS, VS, AVIC
The VS AVIC, in cooperation with State animal health authorities, is responsible for coordination the field activities surrounding the emergency response to BSE. The AVIC assembles the local VS staff to initiate activities outlined in the BSE Red Book including tracing the progeny and herdmates of the suspect animal and beginning an epidemiologic investigation. The VS AVIC coordinates with the State Veterinarian to quarantine the suspect animal's herd of origin. The State has the authority to order a routine quarantine for a neurological disease. The BSE Response Team surveyed every State to determine if they would utilize this authority in the event that NVSL identifies a presumptive diagnosis of BSE. All States responded that they would issue a quarantine.
BSE Response Team
The BSE Response Team leaders will notify each team member and instruct them to assemble in the Situation Room at APHIS headquarters in Riverdale, MD. The Team leaders are responsible for ensuring that all of the Team's duties are fulfilled. It is their responsibility to ensure that the technical information and expert recommendations reach the decisionmakers in a timely fashion. Together with VS Emergency Programs staff, the Team leaders will obtain APHIS, VS administrative support staff in Riverdale, MD, to ready the room for use as BSE headquarters. The Team will begin gathering and assembling information from APHIS and FSIS region and field staff. The Team will pull the draft documents from the third section in the Team leaders manuals and begin filling in current information as it becomes available.
Public Notification
Should NVSL receive notice from CVL confirming a case of BSE, the next level of notification is activated. Each player will follow the same notification protocol as described above for preliminary notification to confirm the diagnosis of a case of BSE.
BSE Response Team
The BSE Response Team will complete the informational memorandum for the Secretary. The Team will prepare the letter to the Office of International Epizootics (OIE), the international animal health organization, for signature by the APHIS, VS Deputy Administrator. OIE requires that all countries submit official notification within 24 hours of confirming a diagnosis of BSE.
The BSE Response Team and the office of the APHIS, VS Deputy Administrator would coordinate a teleconference to inform all APHIS regional directors and AVIC'S.
The BSE Response Team and the office of the FSIS, OPHS Deputy Administrator would coordinate a teleconference to inform all regional and field FSIS offices.
The BSE Response Team would coordinate a teleconference to notify other Federal agencies.
The BSE Response Team would coordinate a teleconference to notify key industry/consumer representatives.
The BSE Response Team and APHIS International Services would notify foreign embassies.
The BSE Response Team would establish a toll-free 800 telephone line for industry representatives, reporters, and the public.
The BSE Response Team would coordinate with APHIS Legislative and Public Affairs and USDA office of Communications to issue a press release the day the diagnosis is confirmed. The press release would announce a press conference to be held the morning after the diagnosis is confirmed...
THE END
From: Terry S. Singeltary Sr. (216-119-138-129.ipset18.wt.net)
Subject: Emergency Operations...BSE Red Book
Date: March 13, 2000 at 1:30 pm PST
BSE Red Book 2.1-35
7.0 Emergency Operations
The section below would be implemented only after a first case of BSE is confirmed in the United States.
7.1 READEO Activation
READEO activation will rarely be necessary for BSE outbreaks. Different from most other foreign animal diseases and infectious diseases, BSE is not a rapidly spreading, acute epizootic; is not thought to be transmitted horizontally between animals within a herd, has an extremely long incubation period, and usually affects only isolated single animals or, at most, a few animals within herds. Because BSE does not spread rapidly, the workload to investigate and manage most outbreaks should not normally exceed the capability of existing local field personnel. READEO activation should be considered only if the particular circumstances of a BSE outbreak warrant. If field personnel feel they are unable to manage a BSE outbreak, they should communicate this to their Regional Director and VS, Emergency Program staff, who will evaluate the need for READEO activation.
7.2 READEO Organization
If READEO is activated, a reference should be made to the revised READEO Manual for further guidance on READEO organization and operations.
7.2.1 Office of the Director
When an animal disease emergency exists, the Task Force Directors are responsible for the READEO activities. The directors immediately move to the location of the outbreak and setup the READEO headquarters. Work is coordinated with State officials of the States involved in the outbreak.
7.2.1.1 State Director--(Note: This is the new designation for the Assistant Director.) Each READEO may have one or more State Directors since each State where the disease outbreak is found will be represented in the READEO by State officials designated by the State Veterinarian.
7.2.1.2 Emergency Program Officer--This individual, designated by the Chief Staff Veterinarian of VS, Emergency Programs, provides liaison between the READEO and the Emergency Programs at APHIS headquarters.
7.2.1.3 Public Affairs Officer--The Public Affairs Officer plans, develops, supervises, and maintains information activities for the READEO.
7.2.1.4 Legal--The Legal Advisor provides counsel and assistance to the READEO.
7.2.1.5 Military--The U.S. Armed Forces Command will designate a senior line officer to be the Military Support Officer on the staff of the READEO Task Force Direction The individual is assigned to be the liaison between the Depart-
October 1998
BSE Red Book 2.1-36
ment of Defense and VS, Emergency Programs, and to coordinate needed military assistance during eradication of an FAD outbreak.
7.2.1.6 Meat and Poultry Inspection Operations--The Meat and Poultry Inspection Operations, Food Safety and Inspection Service, will designate personnel to report to the READEO Task Director and to provide liaison between the Task Force and the Meat and Poultry Inspection Operations.
7.2.1.7 Laboratory Coordination--The Laboratory Coordination Officer will advise the READE(3 Director concerning laboratory capabilities and appropriate laboratory examinations to be conducted to provide needed results as rapidly as possible. This individual will assist with interpretation of results.
7.2.2 Administration The Administrative Officer assigned to the READEO will direct and coordinate all facets of general administrative functions. Refer to the revised READEO Manual for a detailed description of the organization and responsibilities.
7.2.3 Field Operations The Field Operations Officer will direct line operations and supervise field personnel in a READEO. Disease investigation, field epidemiology, disease security and personnel security, animal movement control and quarantine enforcement, appraisals of animals and materials, depopulation and disposal, and cleaning and disinfection are among this person's responsibilities.
7.2.4 Technical Support Staff support consists of a technically competent staff designed to act as a resource for the READEO Task Force. Personnel may include but are not limited to individuals who have expertise in the following areas: animal welfare, data systems, disease reporting, economics, environmental impact, epidemiology, evaluation, orientation and training, risk analysis, and wildlife. The staff communicates the needs of the Field Epidemiology Delivery System (FEDS) to the READEO Director as required to maintain an efficient, accurate, up-to-date FEDS.
7.2.4.1 Animal Welfare---Animal Welfare Officers must be knowledgeable about current Federal and State animal welfare regulations, humane methods of animal depopulation, and socioeconomic concerns related to animal welfare issues. They advise the technical support staff and field operations concerning current procedures and accepted methods for use in the humane depopulation of livestock and poultry.
7.2.4.2 Wildlife-- Wildlife Officers participate with the Director and other officials of the READEO to establish and carry out wildlife policies and objectives for the emergency animal disease operation. Through familiarity with the topography, wildlife density, susceptible wildlife species, and movements of susceptible wildlife, the Wildlife Officers can review maps and make recommendations concerning areas to be included in the quarantined high-risk and buffer zones. These officers maintain contact with local, State, and Federal wildlife enforcement officers and wildlife biologists. They develop strategies for conducting surveys of susceptible wildlife in the outbreak
October 1998
BSE Red Book 2.1-37
area to determine the incidence of the disease. They direct and coordinate the vaccination and depopulation of wild animals as necessary to eliminate the disease.
7.3 Supplies and Equipment During an outbreak of BSE, supplies and equipment should be obtained through normal procurement procedures. If a READEO is activated, supplies and equipment should be ordered through the READEO Procurement and Supply Officer.
7.3.1 General Field Supplies Guidelines Refer to APHIS Directive 326.1, 10/10/77 and 221.1, 1/29/74.
7.4 Personnel Responsibilities During a BSE outbreak, field personnel should follow instructions issued through the normal chain of command. If a READEO is activated, personnel should refer to the revised READEO Manual for detailed descriptions of individual responsibilities.
7.4.1 Personnel Personnel assigned to the READEO Task Force are individually accountable for equipment and supplies checked out to them. They should order replacement equipment and supplies or return equipment for repairs through the READEO Procurement and Supply Officer. All damages or losses to equipment or vehicles should be reported immediately to the READEO Administrative Officer, and the required forms should be completed and submitted promptly.
7.4.2 Travel Employees of the READEO Task Force are responsible for recording and preparing all travel-related documents. Claims for travel, lodging, per diem, and incidental expenses should be submitted to the READEO Administrative Officer for processing.
7.4.3 Vehicles Employees of the READEO Task Force are responsible for operating, cleaning, and performing routine maintenance of assigned vehicles. They also are responsible for recording mileage, expenses, and services. Required reports are to be submitted to the READEO Vehicle Officer.
7.4. 4 Clothing Employees of the READEO Task Force are issued protective clothing to wear when entering a premises where BSE has been diagnosed or is suspected. Clean clothing should be worn on each premises. Employees are responsible for laundering the clothing before reusing it.
October 1998
BSE Red Book 2.1-38
In a large task-foree operation, arrangements may be made for a commercial laundry service to handle the clothing. If it is possible and practical, all clothing should be labeled to identify the employees to whom it is assigned.
7.4.5 Miscellaneous Responsibilities Employees are responsible for conducting their assigned tasks in a professional manner. Complaints concerning task force employees should be directed to the READEO Director for resolution or appropriate action.
All animals, products, and materials to be destroyed because of BSE should be appraised according to 9 CFR 53.3 and appropriate State regulations.
7.5.1 Appraisal Teams Appraisals must represent the interests of the owner, the State, and the Federal Government and be consistent with fair market values. If State authorities approve, State and Federal interests may be represented by a VS employee alone. Owners may, at their discretion and expense, employ a professional appraiser to advise them or to act as their agent. Either the owner or the owner's agent must be present at appraisals. No animals may be destroyed until after the appraisal forms are signed by the owner or the owner's agent. Appraisers should be certain that the owner or the owner's agent is aware of the indemnity form's clause concerning liens and mortgages.
When the number of animals to be destroyed is small, and the total value of animals, products, and materiais is low, APHIS field personnel may negotiate the appraised value with the animal's owner without assistance from a professional appraiser. The appraised value of a BSE suspect should be the slaughter value of the animal, taking into account any existing defects or diseases that would affect the slaughter value but ignoring those signs that caused the animal to be classified as a BSE suspect. If field personnel are in doubt concerning the need to use a professional appraiser, they should consult their supervisor or VS, Emergency Programs staff.
If a determination is made that healthy progeny, ova, semen, or embryos must be destroyed, they should be appraised at 100 percent of replacement value.
Feeds or feed ingredients located on suspect farms will rarely need to be destroyed. If a determination is made that feeds or feed ingredients must be destroyed (for example, to comply with a policy decision to remove all rendered products from animal feeds), then these materials should be appraised and indemnified according to 9 CFR 53.3.
October 1998
BSE Red Book 2.1-39
7.6 Depopulation Procedures
7.6.1 Factors and Considerations If the owner is agreeable, a humane method of euthanasia of BSE suspects will be necessary to facilitate the accurate diagnosis of the disease problem, to ensure that the suspect animal is not slaughtered or rendered, and to terminate the animal's suffering. Under no circumstances may BSE suspects be sent for slaughter or rendering. Notify FDA, CVM if you suspect that the carcass of a BSE-confirmed animal has moved to rendering or animal feed manufacturing. The VS, Emergency Programs staff, Riverdale, MD, must authorize the use of euthanasia, depopulation, and indemnity payments for READEO operations.
7.6.2 Humane Euthanasia Methods Only experienced veterinarians should perform euthanasia because there are inherent dangers. Precautions should be taken to prevent accidents. Owners should be given a complete explanation of what to expect, and only humane euthanasia methods should be used. Euthanasia should be performed away from public view, and, if possible, the owner should not be present. Euthanized animals must be checked to confirm death. (See VS Memo 583.1, 1992.)
7.6.2.1 Mechanical (Firearms)--Because the only acceptable method for euthanizing an animal by using firearms is to shoot it in the head, and because the animal's brain must be preserved to diagnose BSE, firearms are not an acceptable euthanasia method.
7.6.2.2 Chemicals(Toxic Gas or Lethal Injection)--Follow guidelines established by the American Veterinary Medical Association. When using a regulated controlled substance (e.g., barbiturates), control and administration of the euthanasia agent must be given by a veterinarian having a Drug Enforcement Administration (DEA) number issued by the U.S. Treasury. Control and administration of chemical substances for euthanasia must be authorized by the AVIC unless directed by the VS Deputy Administrator. (See VS Memorandum 583.1, 1992.)
7.6.3 Supervision of Depopulation Field personnel should never perform depopulation or euthanasia without explicit permission from their supervisor or, if appropriate, the READEO Humane and Disposal Officer. (Refer to the revised READEC) Manual.)
7. 6.4 Permits for Movement All BSE suspects may be moved under permit to facilitate medical treatment, euthanasia, necropsy examination, or carcass disposal. Permitted movement will be according to the quarantine restrictions and will be administered by the State or Federal officials.
7. 6. 5 Security Because BSE is neither contagious nor vector borne, strict disease security measures are not necessary. Personnel should observe normal disease security measures that are standard procedure for all farm visits. The READEO's Security and Disease Prevention Officer has the responsibility for establishing biosecurity measures.
October 1998
BSE Red Book 2.1-40
7.7 Disposal Under no circumstances may BSE suspects be sent to slaughter or rendering. Notify FDA, CVM if you suspect that the carcass of a BSE-confirmed animal has moved to rendering or animal feed manufacturing. Field personnel should arrange for the carcass to be transported to and examined by a qualified veterinary pathologist or field veterinary medical officer. After the pathologic examination has been completed and the necessary diagnostic specimens have been obtained, field personnel should arrange for disposal of the carcass. Before a method of disposal is selected, there are many factors that must be considered, and often other State and Federal agencies must be consulted. The environmental and legal impacts of the operation must be considered. Upon recommendation of the State or Federal agencies, VS may consider other disposal methods.
7.7.1 Incineration Incineration, although more expensive than burial, is the preferred disposal method for BSE-suspect carcasses. Federal, State, and local environmental regulations may restrict the use of this method and permits may be necessary. As soon as BSE suspects are reported to APHIS, field personnel should investigate the location and availability of incinerators of sufficient size to process a bovine carcass. Institutions likely to have incinerators include State and university diagnostic laboratories, waste contractors, large municipalities, and private industries. Ideally, the diagnostic laboratory where the pathologic examination was done will have incineration facilities. The BSE-suspect carcass disposal is APHIS' responsibility (not the diagnostic laboratory's). Field personnel should arrange for transportation and final disposal of the suspect carcass and should inform their supervisors and/or the READEO Humane and Disposal Officer of these arrangements.
Personnel should be aware that some laboratories dispose of carcasses by rendering and should specifically inquire if this is the case. CNS suspects should be incinerated or held from rendering until a diagnosis of BSE can be ruled out. Under no circumstances may BSE suspects be sent to slaughter or rendering. Notify FDA, CVM if you suspect that the carcass of a BSE-confirmed animal has moved to rendering or animal feed manufacturing.
Field personnel should be prepared to accompany the carcass from the farm of origin to the diagnostic laboratory and then to the disposal site if any doubt exists concerning the final disposal method.
7.7.2 Burial If there are no other avenues for carcass disposal, burial of BSE-suspect carcasses may be an acceptable disposal method. APHIS field personnel should inquire with environmental authorities concerning Federal, State, and local regulations that may impose restrictions on this method.
The burial site may be on the affected farm, at the diagnostic laboratory where the carcass is examined, or in a local landfill. The site should be inaccessible to animals, removed from populated areas, not used for agricultural purposes, clearly marked, and properly protected.
October 1998
BSE Red Book 2.1-41
Burial sites should also be located a sufficient distance from underground utility lines, septic systems, water wells, and surface water. Local environmental or public works officers may be helpful in locating a satisfactory site.
Field personnel should consult with their supervisors and/or the READEO Environmental Impact Officer before digging. Burial trenches are normally at least 9 feet deep with floor dimensions of 7 by 2 feet per adult bovine carcass. Carcasses should be covered with at least 6 feet of soil. This soil should not be tightly packed because gas formation may cause a tightly packed trench to crack and leak.
7.7.3 Rendering Because BSE is spread by rendered animal protein, BSE-suspect and confirmed carcasses must not be rendered, unless the rendered material is incinerated. Notify FDA, CVM if you suspect that dead BSE animals or carcasses have moved to rendering or animal feed manufacturing.
7.7.4 Other Disposal Methods The AVIC, the State animal health officials, and the READEO Director may recommend other methods of disposal to the Deputy Administer, VS, for approval (9 CFR 53.4). Options for disposal must be discussed and approved by VS, Emergency Programs staff and must comply with all State and local Environmental Protection Agency regulations.
7.8 Cleaning and Disinfecting (C&D)
Although BSE is neither contagious nor vector borne, appropriate C&D is required to prevent farm-to-farm transmission of most other infectious diseases. Field personnel must remember, however, that at the time they are requested to euthanize a BSE-suspect animal, a confirmed diagnosis of BSE will not be available. Signs compatible with BSE may be caused by numerous infectious diseases and many BSE-suspect animals will, in fact, have some other disease. Although the C&D of items such as manure, bedding, feed, stalls, halters, milking machines, and other supplies and equipment that have been in contact with BSE suspects is not specifically necessary to control BSE, C&D is still advisable to control other diseases that may be present.
7.8.1 Procedures for Cleaning and Disinfecting
7.8.1.1 Premises and Items--Field personnel are not responsible for C&D of premises such as barns, stalls, and animal pens unless invasive diagnostic procedures (such as a necropsy examination or the removal of the suspect animal's brain) were performed on the premises. If possible, field personnel should avoid doing such procedures on the farm. If circumstances require that such procedures must be done on the farm, personnel should clean and disinfect the immediate area after completing the work.
7.8.1.2 Vehicles--Vehicles used to transport personnel to affected premises should be kept clean, and normal precautions against the farm-to-farm spread of any disease should be observed.
October 1998
BSE Red Book 2.1-42
7,8.1.3 Carriers--Thoroughly clean trucks and trailers transporting BSE suspects. Manure and bedding may be disposed of by any environmentally accepted method such as spreading on fields or composting. After conveyances have been thoroughly cleaned, disinfectant should be sprayed on the sides and floor of the truck bed.
7.8.1.4 Livestock Markets--The risk of BSE transmission at livestock markets is negligible. If a BSE suspect is found at a livestock market, it should be managed the same as if it were found at a farm. Because of the high risk of transmission of diseases other than BSE, invasive diagnostic procedures, such as a necropsy examination or removal of the suspect animal's head, should not be performed at livestock markets. Due to the recent research findings concerning maternal transmission, any pens or areas in which calving occurs should be thoroughly cleaned and disinfected.
Cleaning and disinfection is not necessary to prevent the spread of BSE. However, the C&D procedures are recommended to prevent the spread of other diseases from pens or buildings where BSE suspects were held. 7.8.1.5 Slaughter Plants--Because BSE is spread by rendered animal protein in cattle feeds, BSE suspects must not be slaughtered nor rendered. If a BSE suspect is found at a slaughter plant, it should be managed similarly to finding a suspect at a farm.
7.8.2 Approved Disinfectants Field personnel should use professional judgment in the choice of a disinfectant. Preferred disinfectants to inactivate the BSE agent include 1N sodium hydroxide solution or sodium hypochlorite solution containing 2 percent chlorine (1 hour exposure at 20 %C [68 %F]). This should be used whenever there is reason to strongly suspect that BSE is in fact the cause of the suspect animal's disease. Such reasons include previously confirmed BSE in the geographic area or signs more compatible with BSE than with any other neurologic disease.
If the suspect animal's signs are more compatible with diseases such as rabies or listeriosis, then a phenolic disinfectant such as "One Stroke" may be preferable. (Refer to appendix A Survival of BSE Agent and sec. 1.4.4.)
7. 8. 3 Precautions All disinfectants are hazardous to human beings, animals, and the environment. Label directions should be carefully read and followed. Many disinfectants, including sodium hypochlorite solution, are also corrosive and should be used with caution on metal and other corrodible materials. Thorough rinsing is necessary if corrosive disinfectants are used on metallic items.
Disinfectants, especially in concentrated form, may irritate skin, eyes, and respiratory systems. Protective equipment such as appropriate clothing, rubber boots, rubber gloves, mask and goggles should be worn during mixing and application of disinfectants. If areas of the body are exposed to a disinfectant, they should be washed thoroughly with water. Employees should notify their supervisor and their Health and Safety Officer if excessive human or animal exposure to disinfectants occurs or if there is accidental release into the environment.
October l998
BSE Red Book 2.1-43
Field personnel should use normal hygienic procedures (such as washing and disinfecting boots and removing the outer layer of clothing) when leaving the farm. Unless the disease problem is noncontagious, personnel should not travel to other livestock premises for the duration of that day.
7.9 Vector Control
Current scientific data indicate that BSE is not spread by vectors.
7.10 Disease Prevention and Philosophy
The goal of disease prevention and control is to confine the occurrence of BSE to as few herds as possible and to prevent recycling of the BSE agent in the ruminant food supply. If undiagnosed cases are rendered and included in ruminant rations, the long incubation period may allow many animals to be exposed.
Action should be taken immediately after the detection and confirmation of BSE to initiate an extensive epidemiologic investigation to determine the source and extent of the disease, to stop the spread, and to eradicate the disease.
7.10.1 Philosophy--Immediate action should be taken to prevent contamination of the animal food supply by prohibiting rendering of any infected or suspect bovine carcasses. In addition, care should be taken to monitor those animals born and raised in affected herds and to prevent their becoming a source of infection to other herds.
7.10.2 Agent Spread--Epidemiologic evidence indicates that the primary route of BSE transmission is through the feeding of contaminated meat and bone meal that has been manufactured using scrapie infected sheep carcasses or BSE infected bovine carcasses. Recent research findings suggest that maternal transmission may occur at a rate of approximately 1 percent in some species. It is believed that this route of transmission is not significant enough to maintain an epidemic. Cases of apparent maternal transmission have also been identified in captive exotic ruminants.
7.10.3 Control of Products and Conveyances--Carcasses of BSE suspects should be incinerated. Carcasses must not be rendered and incorporated in animal feed. If carcasses are transported for disposal, conveyances should be cleaned and disinfected after use with either a sodium hypochlorite solution (2 percent available chlorine) or 1 N lye (sodium hydroxide solution).
7.10.4 Control of Biologics and Drugs--Although no documented cases of BSE have resulted from the use of biologics derived from bovines, tissues from suspect or exposed animals must not be used for the production of biologics and drugs.
The agents responsible for causing the transmissible spongiform encephalopathies are highly resistant to normal inactivation processes. Careful selection of source materials is the best way to secure maximum safety of ingredients or reagents of bovine origin used in the manufacture of biologics or other medicinals. Factors that should be considered are the age of the animals, exposure to the agent, and the tissue or organ from which the product is derived.
October 1998
BSE Red Book 2.1-44
7.10.5 Wild Birds, Wind and Insects---Wild birds, wind and insects are not known factors in the spread of BSE.
7.10.6 Rodents--Rodents are not known factors in the spread of BSE. 7.10.7 Hunting--Restrictions on the hunting of wild animals are not necessary to prevent BSE.
7.10.8 Exhibitions--Cancelling scheduled exhibitions is not necessary.
7.10.9 Rendering Trucks and Drivers--The carcasses from BSE suspects must not be rendered. If any rendering truck is used to transport a suspect, it should be cleaned, washed, and disinfected as above. (Refer to appendix A--Agent Survival and sec. 7.8.2--Disinfectants.)
7.10.10 Treatment--Currently there is no known treatment for BSE.
7.10.11 Prevention--Suspects and animals confirmed to have BSE must not be rendered. Producers, feed mills, and rendering establishments should adhere to U.S. State and local rendering policies and FDA regulations concerning the feeding of rendered animal protein to ruminants. Because of the possibility that some transmissible spongiform encephalopathies may be transmitted at the time of parturition, precautions should be taken to prevent exposure of healthy animal to placenta and reproductive fluids. Importation of live animals and animal products from countries with BSE or having high risk factors for BSE should be restricted based upon scientific risk assessment.
7.10.11.1 Immunization--The agent that causes BSE elicits no detectable immune response in the host. Therefore, vaccination is not a viable option. There is no vaccine currently developed for BSE or other TSE's.
7.10.11.2 Sanitation--Although it is unknown whether a contaminated environment plays any role in the spread of BSE, it is suggested that pens having contained BSE-infected animals be cleaned and disinfected. The disinfectants o choice are sodium hydroxide (lye) and sodium hypochlorite, in infected herds it is also advisable that all placentas be removed promptly and buried or incinerated. The calving pens also should be cleaned and disinfected.
7.10.113 Producer Defense---The most effective way to prevent an introduction of BSE into a herd is not to feed ruminant byproducts to ruminants. As of August 4, 1997, the FDA has a ban in place which prohibits the feeding of most mammalian proteins to ruminants.
7.11 Records Maintenance in a Foreign Animal Disease Outbreak
The APHIS FEDS will be used by the READEO to record information. FEDS a computerized network designed to transmit accurate information rapidly during any emergency disease outbreak. The use of FEDS will allow the READEO to direct its attention to the minute-to-minute business of containing and eradicating the disease.
For an accurate record of the activities, all field supervisors in a READEO task force should maintain a diary. Activities and observations should be recorded in the diary when they occur. Date all documents and enter events by time and date to show a correct chronology.
Enter events as they occur in the diary as well. An accurate history is of considerable value in developing policies and plans for future disease-eradication
October 1998
BSE Red book 2.1-45
programs, and it may be important if there is litigation. A diary will be helpful for day-to-day administration of funds, personnel, and equipment. It is also useful as a later reference in preparing reports and summaries of activities.
7.11.1 Daily Reports Submit daily reports of significant activities to the READEO Director and the VS, Emergency Programs staff Riverdale, MD. (Refer to appendix F for current telephone listings.) Include the following as part of the historical file of an outbreak:
*Maps showing premises where BSE-infected animals were found;
*Inventory of feeds and feed sources;
*Origin of BSE-suspect and confirmed animals;
*Public information material distributed, newspaper clippings; and,
Administrative reports to support the expenditure of funds, utilization of personnel and equipment, and disposition of excess materials and equipment at the end of the program.
7.11.2 Distribution The VS, Emergency Programs staff will distribute reports of significant activities to all AVIC's, State cooperators, and industry cooperators at least weekly. As soon as significant events occur, Emergency Programs will inform all APHIS
will inform all APHIS headquarters units through normal reporting channels. Emergency Programs also will immediately report any significant events to the Deputy Administrator, VS, who will immediately advise the APHIS Administrator, especially of legal or politically important events. A weekly summary report of control and eradication activities will be provided to the APHIS Administrator and the Deputy Administrator, VS. See BSE Response Plan, communications section.
7.11.3 Disposition Records should be maintained until a historical account of the program has been prepared and all pertinent information has been gleaned from the records.
Furthermore, all records should be maintained if there may be legal action pending as a result of the program activities. Usually, administrative records are maintained a minimum of 3 years for audit purposes.
END...TSS
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USDA Bovine Spongiform Encephalopathy BSE, Scrapie, CWD, Testing and Surveillance 2022 A Review of History
Greetings, i lost my Mom to the Heidenhain Variant of Creutzfeldt Jakob Disease hvCJD way back on December 14, 1997. At the same time, we were watch Oprah Winfrey defend herself against the Cattle Industry in Texas about that Mad Cow Hamburger and those staggering cows she showed on her T.V. I said right then we had a problem here in the USA, and nobody was taking it seriously.
Since the, I have followed the science daily on TSE Prion, and every year around this time, i like to put together an annual report of sorts for BSE, Scrapie, CWD, now CPD i.e Camel Prion Disease, and the different Human TSE Prion strains. sometimes i may report on them individually, and sometimes just one report that includes all of it.
I sit here today in 2022 and say the same thing i said back from 1998 and forward, and would kindly like to show you. In fact, the Texas Mad Cow (the second one, the first one was sent off to be rendered without any testing at all, but this is the second one i speak of now), this cow Texas tried to cover up with BSE testing that was the least likely to find BSE, and in fact, i was so concerned, i wrote the OIG Office of Inspector General, and i also sent of letters to a bunch of scientist that i was concerned this cow that was negative, was in fact positive. After a year of mad cow wrangling with Scientist and the OIG, that negative mad cow was CONFIRMED AS POSITIVE. I will review Past and Present BSE testing, surveillance, the infamous 'Enhanced BSE Surveillance'' and the Harvard Assessment ONE and TWO, and a failed attempt to combat this deadly zoonotic disease, or was it a successful attempt at failing to find BSE in the USA?
I believe, imo, and through decades of following this daily, that the USDA/APHIS BSE Testing formula was intentionally set up to never detect BSE TSE Mad Cow disease, and this includes Scrapie TSE in Sheep and Goat.
so, let's review the facts.
FIRST, let's review what the EU is doing today, in terms of testing figures, surveillance, and detection in 2021...
The European Union summary report on surveillance for the presence of transmissible spongiform encephalopathies (TSE) in 2021
Published:
30 November 2022
Approved: 3 November 2022
Metadata
EFSA Journal 2022;20(11):7655
Keywords: TSE, BSE, CWD, scrapie, classical, atypical, surveillance
On request from: European Commission Question Number: EFSA‐Q‐2021‐00765
Contact: zoonoses@efsa.europa.eu
Abstract
This report presents the results of surveillance on transmissible spongiform encephalopathies (TSE) in cattle, sheep, goats, cervids and other species, and genotyping in sheep and goats, carried out in 2021 by 27 Member States (MS, EU27), the United Kingdom (in respect of Northern Ireland) (XI), and eight other non‐EU reporting countries: Bosnia and Herzegovina, Iceland, Montenegro, North Macedonia, Norway, Serbia, Switzerland and Turkey.
In total, 1,021,252 cattle were tested by EU27 and XI (−9%, compared with 2020 when data from the United Kingdom were not restricted to Northern Ireland), and 66,121 cattle by eight non‐EU reporting countries,
with two cases of H‐BSE in France and Spain, and four L‐BSE in France (2), Germany and Spain.
In total, 311,174 sheep and 118,457 goats were tested in the EU27 and XI (−6.4% and −1.8%, respectively, compared to 2020 when data from the whole United Kingdom were considered).
In sheep, 551 cases of scrapie were reported by 17 MS and XI: 448 classical scrapie (CS) by six MS [80 index cases (IC) with genotypes of susceptible groups in 97% of the cases], 103 atypical scrapie (AS) (96 IC) by 13 MS and XI.
In the other non‐EU reporting countries, 27,594 sheep were tested with 55 CS and 1 AS in Iceland and 8 AS in Norway.
Ovine random genotyping was reported by nine MS and genotypes of susceptible groups accounted for 7.9%.
In goats, 224 cases of scrapie were reported by six EU MS: 219 CS (30 IC) by six MS, and five AS (5 IC) by three MS.
In total, 5,854 cervids were tested for chronic wasting disease by eight MS; all resulted negative. Norway tested 21,670 cervids with two moose and one red deer positive. In total, 149 animals from four other species tested negative in Finland and Turkey.
© European Food Safety Authority
see full text;
SECOND, let's review the USDA/APHIS et al BSE, Scrapie, and CWD testing, past and present...
USDA BSE Surveillance Information Center
Introduction USDA conducts surveillance for Bovine spongiform encephalopathy (BSE), referred to as "mad cow disease", in cattle to determine if, and at what level, the disease is present in the U.S. cattle population. Our surveillance program allows us to assess any change in the BSE disease status of U.S. cattle, and identify any rise in BSE prevalence in this country. Identifying any changes in the prevalence of disease allows us to match our preventive measures - feed ban for animal health, and specified risk material removal for public health - to the level of disease in U.S. cattle.
It is the longstanding system of interlocking safeguards, including the removal of specified risk materials - or the parts of an animal that would contain BSE - at slaughter and the FDA's ruminant-to-ruminant feed ban that protect public and animal health from BSE.
Why did USDA decrease the number of samples per year in 2006? After the first confirmation of BSE in an animal in Washington State in December 2003, USDA evaluated its BSE surveillance efforts in light of that finding. We determined that we needed to immediately conduct a major surveillance effort to help determine the prevalence of BSE in the United States. Our goal over a 12-18 month period was to obtain as many samples as possible from the segments of the cattle population where we were most likely to find BSE if it was present. This population of cattle was exhibiting some signs of disease. We conducted this enhanced surveillance effort from June 2004 - August 2006. In that time, we collected a total of 787,711 samples and estimated the prevalence of BSE in the United States to be between 4-7 infected animals in a population of 42 million adult cattle. We consequently modified our surveillance efforts based on this prevalence estimate to a level we can monitor for any potential changes, should they occur. Our statistical analysis indicated that collecting approximately 40,000 samples per year from the targeted cattle population would enable us to conduct this monitoring.
Why is USDA "only" testing 25,000 samples a year? USDA's surveillance strategy is to focus on the targeted populations where we are most likely to find disease if it is present. This is the most effective way to meet both OIE and our domestic surveillance standards. After completing our enhanced surveillance in 2006 and confirming that our BSE prevalence was very low, an evaluation of the program showed that reducing the number of samples collected to 40,000 samples per year from these targeted, high risk populations would allow us to continue to exceed these standards. In fact, the sampling was ten times greater than OIE standards. A subsequent evaluation of the program in 2016 using data collected over the past 10 years showed that the surveillance standards could still be met with a further reduction in the number of samples collected by renderers and 3D/4D establishments which have a very low OIE point value because the medical history of these animals is usually unknown. Therefore, in 2016, the number of samples to be tested was reduced to 25,000 where it remains today.
How can USDA find every case of BSE in the United States when you are only testing 25,000 animals? The goal of our BSE surveillance program, even under the enhanced program, has never been to detect every case of BSE. Our goal is determine whether the disease exists at very low levels in the U.S. cattle population, and we do this by testing those animals most likely to have BSE. It is the longstanding system of interlocking safeguards, including the removal of specified risk materials - or the parts of an animal that would contain BSE - at slaughter and the FDA's ruminant-to- ruminant feed ban that protect public and animal health from BSE.
Why didn’t USDA continue to test animals at the enhanced surveillance level? USDA's 2004-2006 enhanced surveillance program was initiated in response to the first detection of BSE in the United States and was designed to detect the overall prevalence of the disease in this country. This required a very intensive effort and it allowed us to estimate extremely low prevalence levels of disease. Once that prevalence level was determined, USDA modified its testing levels to monitor any changes in the level of disease. Our current testing of approximately 25,000 targeted animals a year allows USDA to detect BSE at the very low level of less than 1 case per million adult cattle, assess any change in the BSE status of U.S. cattle, and identify any rise in BSE prevalence in this country.
Is USDA's surveillance program a food safety or public health measure? The primary, and most effective, food safety or public health measure is the removal of specified risk materials (SRMs) - or the parts of an animal that would contain BSE - from every animal at slaughter. USDA's BSE surveillance program is not a food safety measure; it is an animal health monitoring measure. However, it does support existing public health and food safety measures. By allowing us to monitor the level of disease in the US cattle population, we can help determine the appropriate level of public health and animal health measures required, and whether they should be increased or decreased.
Why doesn't USDA test every animal at slaughter? There is currently no test to detect the disease in a live animal. BSE is confirmed by taking samples from the brain of an animal and testing to see if the infectious agent - the abnormal form of the prion protein - is present. The earliest point at which current tests can accurately detect BSE is 2 to 3 months before the animal begins to show symptoms, and the time between initial infection and the appearance of symptoms is about 5 years. Therefore, there is a long period of time during which current tests would not be able to detect the disease in an infected animal.
Since most cattle are slaughtered in the United States at a young age, they are in that period where tests would not be able to detect the disease if present. Testing all slaughter cattle for BSE could produce an exceedingly high rate of false negative test results and offer misleading assurances of the presence or absence of disease.
Simply put, the most effective way to detect BSE is not to test all animals, which could lead to false security, but to test those animals most likely to have the disease, which is the basis of USDA's current program.
What animals are USDA testing in the surveillance program? These are random samples at slaughter, aren't they? No. USDA's BSE surveillance program is specifically targeted to the population most likely to have the disease, if it is present. This population is NOT clinically healthy animals that would be presented for slaughter. Rather, it includes animals that have some type of abnormality, such as central nervous system signs; non-ambulatory, or a "downer"; emaciated; or died for unknown reasons. Because these animals would not pass the required ante-mortem inspection requirements at slaughter for human consumption, we collect the majority of our samples at facilities other than slaughter facilities - at rendering or salvage facilities, on-farm, at veterinary clinics or veterinary diagnostic laboratories. With this targeted approach, we can monitor the presence of disease in the US cattle population in a much more efficient and meaningful way. The key to surveillance is to look where the disease is going to occur.
Key Points: BSE Ongoing Surveillance Plan Note: This Fact Sheet is based on the USDA Animal and Plant Health Inspection Service (APHIS) Bovine Spongiform Encephalopathy (BSE) Ongoing Surveillance Plan, July 20, 2006. To learn more, read the complete BSE Ongoing Surveillance Plan (PDF, 187 KB).
KEY POINTS In addition to a stringent feed ban imposed by the Food and Drug Administration in 1997 as well as the removal of all specified risk material which could harbor BSE, USDA has a strong surveillance program in place to detect signs of BSE in cattle in the United States. In fact, we test for BSE at levels greater than World Animal Health Organization standards. The program samples approximately 25,000 animals each year and targets cattle populations where the disease is most likely to be found. The targeted population for ongoing surveillance focuses on cattle exhibiting signs of central nervous disorders or any other signs that may be associated with BSE, including emaciation or injury, and dead cattle, as well as non-ambulatory animals. Samples from the targeted population are taken at farms, veterinary diagnostic laboratories, public health laboratories, slaughter facilities, veterinary clinics, and livestock markets.
USDA's National Veterinary Services Laboratories (NVSL) in Ames, IA, along with contracted veterinary diagnostic laboratories, use rapid screening tests as the initial screening method on all samples. Any inconclusive samples undergo further testing and analysis at NVSL.
NOT A FOOD SAFETY TEST BSE tests are not conducted on cuts of meat, but involve taking samples from the brain of a dead animal to see if the infectious agent is present. We know that the earliest point at which current tests can accurately detect BSE is 2-to-3 months before the animal begins to show symptoms. The time between initial infection and the appearance of symptoms is about 5 years. Since most cattle that go to slaughter in the United States are both young and clinically normal, testing all slaughter cattle for BSE might offer misleading assurances of safety to the public.
The BSE surveillance program is not for the purposes of determining food safety. Rather, it is an animal health surveillance program. USDA's BSE surveillance program allows USDA to detect the disease if it exists at very low levels in the U.S. cattle population and provides assurances to consumers and our international trading partners that the interlocking system of safeguards in place to prevent BSE are working.
The safety of the U.S. food supply from BSE is assured by the removal of specified risk materials - those tissues known to be infective in an affected animal - from all human food. These requirements have been in place since 2004.
ONGOING BSE SURVEILLANCE PROGRAM SUMMARY USDA's BSE surveillance program samples approximately 25,000 animals each year and targets cattle populations where the disease is most likely to be found. The statistically valid surveillance level of 25,000 is consistent with science-based internationally accepted standards. This level allows USDA to detect BSE at the very low level of less than 1 case per million adult cattle, assess any change in the BSE status of U.S. cattle, and identify any rise in BSE prevalence in this country.
The targeted population for ongoing surveillance focuses on cattle exhibiting signs of central nervous disorders or any other signs that may be associated with BSE, including emaciation or injury, and dead cattle, as well as nonambulatory animals. Samples from the targeted population are taken at farms, veterinary diagnostic laboratories, public health laboratories, slaughter facilities, veterinary clinics, and livestock markets.
USDA's National Veterinary Services Laboratories (NVSL) in Ames, IA, along with contracted veterinary diagnostic laboratories, will continue to use rapid screening tests as the initial screening method on all samples. Any inconclusive samples will be sent to NVSL for further testing and analysis. USDA's surveillance program uses OIE's weighted surveillance points system, which was adopted in May 2005 and reflects international scientific consensus that the best BSE surveillance programs focus on obtaining quality samples from targeted subpopulations rather than looking at the entire adult cattle population.
The number of points a sample receives correlates directly to an animal's clinical presentation at the time of sampling. The highest point values are assigned to those samples from animals with classic clinical signs of the disease. The lowest point values correspond to clinically normal animals tested at routine slaughter.
The goal of this weighted approach is to ensure that countries sample those cattle populations where the disease is most likely to be found. This system is not different from USDA's previous BSE surveillance approach, it is simply a different method for evaluating surveillance programs. Both approaches target those cattle populations where BSE is most likely to be found. The OIE is simply assigning point values to different categories of animals.
USDA has been targeting these subpopulations since BSE surveillance was initiated in 1990, and will continue to do so under the OIE weighted approach. Under the OIE guidelines, points compiled over a period of 7 consecutive years are used as evidence of adequate surveillance. At the current ongoing level of surveillance, the United States will far exceed OIE guidelines under the point system.
MY BULLSHIT METER PEGGED OUT ON THE ABOVE BSe by USDA et al!
COMPARING the EU, to the USA, on BSE Surveillance and Testing, you can see for yourself why the EU is finding more, because they are testing more, compared to USA.
''USDA's 2004-2006 enhanced surveillance program was initiated in response to the first detection of BSE in the United States and was designed to detect the overall prevalence of the disease in this country.''
I would kindly like to remind everyone of this very important document;
BSE research project final report 2005 to 2008 SE1796 SID5
I was very concerned of the BSE testing and even discussed this with Bio-Rad et al;
Audit Report
Animal and Plant Health Inspection Service
Bovine Spongiform Encephalopathy (BSE) Surveillance Program – Phase II
and
Food Safety and Inspection Service
Controls Over BSE Sampling, Specified Risk Materials, and Advanced Meat Recovery Products - Phase III
Report No. 50601-10-KC January 2006
Finding 2 Inherent Challenges in Identifying and Testing High-Risk Cattle
Still Remain Our prior report identified a number of inherent problems in
identifying and testing high-risk cattle.
snip...
BIO-RAD
> > -------- Original Message --------
> > Subject: USA BIO-RADs INCONCLUSIVEs
> > Date: Fri, 17 Dec 2004 15:37:28 -0600
> > From: "Terry S. Singeltary Sr."
> > To:
> >
> >
> >
> > Hello xxxx and Bio-Rad,
> >
> > Happy Holidays!
> >
> > I wish to ask a question about Bio-Rad and USDA BSE/TSE testing
> > and there inconclusive. IS the Bio-Rad test for BSE/TSE that
complicated,
> > or is there most likely some human error we are seeing here?
> >
> > HOW can Japan have 2 positive cows with
> > No clinical signs WB+, IHC-, HP- ,
> > BUT in the USA, these cows are considered 'negative'?
> >
> > IS there more politics working here than science in the USA?
> >
> > What am I missing?
> >
> >
> >
> > -------- Original Message --------
> > Subject: Re: USDA: More mad cow testing will demonstrate beef's safety
> > Date: Fri, 17 Dec 2004 09:26:19 -0600
> > From: "Terry S. Singeltary Sr."
> > snip...end
> >
> >
> > Experts doubt USDA's mad cow results
>
>
>
> snip...END
>
> WELL, someone did call me from Bio-Rad about this,
> however it was not xxxxxx xxxxx.
> but i had to just about take a blood oath not to reveal
> there name. IN fact they did not want me to even mention
> this, but i feel it is much much to important. I have omitted
> any I.D. of this person, but thought I must document this ;
>
> Bio-Rad, TSS phone conversation 12/28/04
>
> Finally spoke with ;
>
>
> Bio-Rad Laboratories
> 2000 Alfred Nobel Drive
> Hercules, CA 94547
> Ph: 510-741-6720
> Fax: 510-741-5630
> Email: XXXXXXXXXXXXXXXXXX
>
> at approx. 14:00 hours 12/28/04, I had a very pleasant
> phone conversation with XXXX XXXXX about the USDA
> and the inconclusive BSE testing problems they seem
> to keep having. X was very very cautious as to speak
> directly about USDA and it's policy of not using WB.
> X was very concerned as a Bio-Rad official of retaliation
> of some sort. X would only speak of what other countries
> do, and that i should take that as an answer. I told X
> I understood that it was a very loaded question and X
> agreed several times over and even said a political one.
>
> my question;
>
> Does Bio-Rad believe USDA's final determination of False positive,
> without WB, and considering the new
> atypical TSEs not showing positive with -IHC and -HP ???
>
> ask if i was a reporter. i said no, i was with CJD Watch
> and that i had lost my mother to hvCJD. X did not
> want any of this recorded or repeated.
>
> again, very nervous, will not answer directly about USDA for fear of
> retaliation, but again said X tell
> me what other countries are doing and finding, and that
> i should take it from there.
> "very difficult to answer"
>
> "very political"
>
> "very loaded question"
>
> outside USA and Canada, they use many different confirmatory tech. in
> house WB, SAF, along with
> IHC, HP, several times etc. you should see at several
> talks meetings (TSE) of late Paris Dec 2, that IHC- DOES NOT MEAN IT IS
> NEGATIVE. again, look what
> the rest of the world is doing.
> said something about Dr. Houston stating;
> any screening assay, always a chance for human
> error. but with so many errors (i am assuming
> X meant inconclusive), why are there no investigations, just false
> positives?
> said something about ''just look at the sheep that tested IHC- but were
> positive''. ...
>
>
> TSS
>
> -------- Original Message --------
> Subject: Your questions
> Date: Mon, 27 Dec 2004 15:58:11 -0800
> From: To: flounder@wt.net
>
>
>
> Hi Terry:
>
> ............................................snip Let me know your phone
> number so I can talk to you about the Bio-Rad BSE test.
> Thank you
>
> Regards
>
>
>
> Bio-Rad Laboratories
> 2000 Alfred Nobel Drive
> Hercules, CA 94547
> Ph: 510-741-6720
> Fax: 510-741-5630
> Email: =================================
>
>
> END...TSS
>
>
Audit Report
Animal and Plant Health Inspection Service
Bovine Spongiform Encephalopathy (BSE) Surveillance Program – Phase II
and
Food Safety and Inspection Service
Controls Over BSE Sampling, Specified Risk Materials, and Advanced Meat Recovery Products - Phase III
Report No. 50601-10-KC January 2006
Finding 2 Inherent Challenges in Identifying and Testing High-Risk Cattle
Still Remain Our prior report identified a number of inherent problems in
identifying and testing high-risk cattle.
snip...
NOW, Back to this very important document, and what i suspected back then was suspicious, and sure enough, years later, i find this document;
BSE research project final report 2005 to 2008 SE1796 SID5
Executive Summary
7. The executive summary must not exceed 2 sides in total of A4 and should be understandable to the intelligent non-scientist. It should cover the main objectives, methods and findings of the research, together with any other significant events and options for new work.
Studies of Bovine Spongiform Encephalopathy (BSE), carried out in the UK, showed it to be a single strain of prion disease based on histopathological (Simmons et al., 1996) and transmission data (Bruce et al., 1992 ). First reported in the 1980s (Wells et al., 1987) there appears to have been little change in the characteristics of the disease throughout the epidemic and BSE maintains a distinct molecular profile even following cross species transmission. However, during surveillance programmes in Europe and in North America two other distinct isolates of bovine prion disease have come to light, H and L type, so-called to reflect their unique molecular profiles (Yamakawa et al., 2003; Biacabe et al., 2004).
Reports were also emerging of atypical forms of scrapie that were distinct from classical scrapie isolates and were less easily recognised by the then current diagnostic tests (Benestad et al., 2003; Buschman et al., 2004). This led to concerns that cattle could also harbour a prion disease that was not detected by the current diagnostic tests for BSE. Importantly, approximately 15-20% of the clinical cases submitted for investigation were indeed negative and this proportion of negative cattle did not appear to vary despite increasing awareness of BSE clinical signs by the farming and veterinary community. While there maybe other explanations for this discrepancy (McGill et al., 1993), another underlying undiagnosed prion disease of cattle distinct from classical BSE could not be ruled out.
The study reported here investigated a small number of these BSE negative clinical cases by using more sensitive and modified diagnostic tests for abnormal PrP.
The majority of the cases that we studied were negative by all the tests employed and based on this observation we conclude that there was not a simultaneous epidemic of another form of bovine prion disease. However, we observed a number of classical cases that were missed prior to the advent of sensitive and rapid diagnostic tests and this provides an estimate of the number of cattle that were mis-diagnosed before 2000. In addition, we observed a few rare cases where the diagnostic tests were not in agreement and these cases were investigated further. One of these unusual samples emerged as a case of idiopathic bovine neuronal chromatolysis (IBNC).
During the study we also reported the first H-type BSE case in the UK (Terry et al., 2007).
snip...
Scientific Objectives as prescribed in the project:
All of the objectives have been met and are described in detail below. Three annexes accompany this report, one with the figures for the results below and two papers for submission to peer-reviewed journals.
Objective 1: To determine the variation of PK sensitivity of bovine PrPc from uninfected cattle brains and compare with bovine PrPsc from classical cases of BSE in order to set thresholds for negative, weak and strong positive values in commercially available rapid diagnostic tests.
Objective 2: Determine whether there are a greater proportion of bovine brain samples positive for the rapid diagnostic tests (hereby called reactors) in the clinically-suspect, negative subset of cattle than in healthy negative cattle. (True positives will be determined on the basis of evaluation by IHC but should be strongly positive in both the rapid diagnostic tests).
Objective 3: Determine whether the phenotypic and molecular characteristics of PrP from cattle identified in 2 are distinct from normal PrPc and from bovine PrPsc normally associated with classical BSE.
Studies of Bovine Spongiform Encephalopathy (BSE), carried out in the UK, showed it to be a single strain of prion disease based on histopathological (Simmons et al., 1996) and transmission data (Bruce et al., 1992). First reported in the 1980s (Wells et al., 1987) there appears to have been little change in the these characteristics of the disease throughout the epidemic; BSE also appear to maintain a distinct molecular profile in cattle and even when experimentally (or naturally) transmitted to other species such as humans and cats. However, during surveillance programmes in Europe, Japan and in North America, two other distinct isolates of bovine prion disease have come to light, H and L type, so-called to reflect their unique molecular profiles (Yamakawa et al., 2003; Biacabe et al., 2004). In the late 1990’s, a novel prion disease was discovered in sheep (Benestad et al., 2003; Buschman et al., 2004); this Nor98 or atypical scrapie is widespread in Europe but had previously been missed by histopathological or immunohistological examination. This led to concerns that cattle could also harbour a prion disease that, unlike H- and L-type BSE, was not detected by the current diagnostic tests for BSE. Importantly, approximately 15-20% of the clinical cases submitted for investigation were indeed negative and this proportion of negative cattle did not appear to vary despite increasing awareness of BSE clinical signs by the farming and veterinary community. While there maybe other explanations for this discrepancy (McGill et al., 1993), another underlying undiagnosed prion disease of cattle distinct from classical BSE could not be ruled out. The study reported here investigated a small number of these BSE negative clinical cases by using more sensitive and modified diagnostic tests for abnormal PrP. The majority of the cases that we studied were negative by all the tests employed and based on this observation we conclude that there was not a simultaneous epidemic of another form of bovine prion disease. However, we observed a number of cases of BSE in this “BSE negative” sub-set that were missed prior to the advent of more sensitive and rapid diagnostic tests and this provides an estimate of the number of cattle that were mis-diagnosed before 2000. In addition, we observed a few rare cases where the diagnostic tests were not in agreement and these cases were investigated further. One of these unusual samples emerged as a case of idiopathic bovine neuronal chromatolysis (IBNC) (Jeffrey & Wilesmith, 1992; 1996; Jeffrey et al., 2009). During the study we also reported the first H-type BSE case in the UK (Terry et al., 2007). Materials and Methods Tissue samples. Test samples: Frozen brain stem from 501 bovine BSE suspects with neurological signs, a) that were negative at the level of the obex for vacuolation by standard histopathological techniques from years 1991-1999 and b) by IHC and diagnostic Bio-Rad PlateliaTM from 2000 onwards. These tissues have been stored at the VLA at –80oC since submission.
Negative controls: Frozen brain stem from 90 cattle investigated as part of the active surveillance programme. These samples were submitted in 2006 to LGC for rapid testing by Bio-Rad TeSeE diagnostic ELISA and were negative. These samples were stored at –80oC prior to testing and were stored for a maximum of 36 months and therefore considerably less time than all experimental samples under investigation.
Cattle with suppurative encephalitis: 10 additional cattle samples were retrieved from the VLA Archive that were negative for BSE but showed signs of suppurative encephalitis and inflammation (lymphocyte cuffing and gliosis). These signs were consistent with listeria infection.
Tests for disease-associated PrP IDEXX BSE Herdchek BSE antigen test kit
All samples were assayed using the IDEXX Herdchek Bovine Spongiform Encephalopathy (BSE) Antigen Test Kit, EIA according to the manufacturer’s instructions and without modification. Briefly, brains were homogenised in the buffer provided by the manufacturer and diluted prior to adding to the seprion (polyanion) coated multiwell plate and incubated prior to washing. The samples were then treated with a conditioning buffer to expose the antigen epitopes. PrPsc was detected by PrP specific antibodies conjugated to horseradish peroxidase and visualised with TMB substrate. Samples were read using a microtitre plate reader (Victor-Perkin-Elmer). The method has no Proteinase K digestion step and has only a mild trypsin treatment that is not required for specificity but aids in the epitope exposure step. The normal curve of negative control samples is provided by the manufacturer and shows the diagnostic cut off value is set higher than most negative controls. The amount of brain added to a single well is approximately 20 mg. Diagnostic Bio-Rad TeSeE EIA
Sample extraction and detection was carried out according to the manufacturer’s instructions for the Bio-Rad TeSeE BSE ELISA. Briefly brain samples were homogenised in buffer provided by the manufacturer and then treated for 10 mins with Proteinase K at 37oC . The PK concentration is not provided by the manufacturer so we refer to it as 4 ul/ml which is the quantity of stock PK to final solution directed by Bio-Rad. A comparison with sigma PK indicated that the concentration is approximately 40 ug/ml. The samples were then precipitated and concentrated by centrifugation. Pellets were reconstituted and diluted in the buffers provided by the manufacturer. The PrPsc was then detected by a sandwich ELISA provided by the manufacturer. Details of the antibodies are not provided. Samples are read using a microtitre plate as above. Cut off values for the ELISA are calculated using the mean of four negative control ODs. The manufacturers indicate that a value of 0.14 should be added to the negative control mean and samples equal to or greater than this value should be further analysed. The amount of brain added to a single well is approximately 65 mg. BioRad TeSeE EIA with reduced PK digestion (0.3 Bio-Rad TeSeE ELISA)
The PrPsc associated with atypical scrapie is believed to be less PK resistant than classical scrapie (Everest et al., 2006). In order to investigate whether an atypical form of BSE in cattle exists biochemically similar to atypical scrapie a modified version of the Bio-Rad TeSeE protocol, using sub-diagnostic levels of Proteinase K (0.3ul/ml), was used. This quantity of PK was arrived at by titration of PK and digestion of PrPc from 47 cattle brains negative for TSEs.
The Bio-Rad TeSeE BSE diagnostic test was used as directed by the manufacturer with the addition of DNAse prior to the Proteinase K (0.3 ul/ml PK) treatment and Pefabloc was added alongside the kit PK stopping solution. The PK dilution for these assays was prepared from a Sigma stock solution and 0.3 units/ml was the equivalent activity as 0.3 ul/ml of Biorad PK. Bio-Rad TeSeE Western Blot
Sample extraction was carried out according to the manufacturer’s instructions (Bio-Rad TeSeE Western Blot) with several modifications. In brief, brain tissue was ribolysed to give 20 % (w/v) homogenate and subsequently incubated with DNAse. The samples were then digested with 0.3, 1, 4 or 20 units/ml PK (Sigma; where units/ml is an in-house nomenclature and 0.3 units/ml is equivalent to 0.3 µl of the Bio-Rad test PK in terms of activity as compared using the TAME test -Pierce) and the reaction stopped with Pefabloc. Following precipitation and centrifugation at 15,000 g for 7 minutes, in accordance with the Bio-Rad TeSeE Western blot protocol, the pellets were re-suspended in Laemmli sample buffer.
For analysis, the supernatants were heated at 100oC for 5 minutes, loaded on a 12% Criterion XT Bis-tris SDS gel (Bio-Rad) and subjected to electrophoresis in XT-MOPS running buffer (Bio-Rad) at 200 V for 50 minutes. Proteins were transferred to a PVDF membrane (Bio-Rad) at 115 V, 60 min using Tris/CAPS transfer buffer (Bio-Rad).
Blots to be evaluated using the Sha31 (Bio-Rad) antibody were incubated for one hour with the blocking solution provided by the manufacturers; and antibodies SAF84 (aa 175-180), P4 (aa 89-104) and FH11 (aa 55-65) using a 5% milk powder in PBS supplemented with Tween 20 (PBST). The membranes were incubated for one hour with the primary antibody and then with goat anti-mouse IgG antibody conjugated to horseradish peroxidase (Bio-Rad) prior to visualization by chemiluminescence (ECL; Amersham). Immunohistochemical analysis Formalin-fixed, paraffin wax-embedded tissue blocks were sectioned at 4mm, collected onto frosted charged slides (GmbH) and melted on at 60°C overnight to improve adhesion. Sections were de-waxed in xylene and alcohol and washing in water. They were subsequently put into 98% formic acid (Merck) for 30 minutes, washed in running tap water for 15 minutes and then fully immersed into citrate buffer (200mM trisodium citrate dehydrate (Sigma), 30mM citric acid (Sigma), pH 6.1) prior to being autoclaved for 30 minutes at 121°C. Endogenous peroxidase activity was quenched using 3% hydrogen peroxide (Sigma) and the sections immersed in purified water and stored at 4°C overnight. After warming to room temperature, non-specific antibody binding sites were blocked using normal goat serum (Vector Laboratories) for 20 minutes. Rat monoclonal anti-PrP R145 (VLA) was diluted to 2mg/ml and applied for one hour at ambient (19°C-24°C) temperature. Biotinylated rabbit-anti-mouse IgG (Vector Laboratories) was diluted appropriately and applied for 30 minutes at ambient temperature. Elite ABC (Vector Laboratories) was prepared according to the manufacturers’ directions and applied for 20 minutes at ambient temperature. Sections were washed between each stage using 5mM tris buffered saline supplemented with Tween-20 (5mM tris, 0.85% NaCl, 0.05% tween-20 (all from Sigma), pH 7.6). Diaminobenzidine tablets (Sigma) were prepared in McIlvanes buffer (200mM disodium hydrogen orthophosphate, 100mM citric acid (both from Sigma), pH 6.4) and applied for 10 minutes at ambient temperature. Sections were counterstained in Mayer’s haematoxylin and “blued” in running tap water, before being dehydrated through three changes each of absolute alcohol and xylene for three minutes each and finally mounted in DPX (Sigma). Definition of terms
Disease associated isoforms of PrP may be distinguished from normal PrP by its increased resistance to Proteinase digestion in immunoblotting or ELISA tests (PrPres), binding to polyanions or labelling with PrP specific antibodies in fixed and treated paraffin-embedded section (PrPd). Included within the operational definition of PrPd are all those detection systems that do not use Proteinase K digestion. The correlation between prion infectivity and PrPres or PrPd is inexact, and infectivity has been dissociated from PrPres or PrPd in several experiments, putatively this is because only a fraction of abnormal PrP isoforms are infectious. We will therefore use operational definitions for detected abnormal PrP forms and PrPsc for the hypothetical infectious sub-population of PrP isoforms detected by bioassay. Results Brains from cattle previously diagnosed as negative for BSE based on histopathological examination were investigated in this study for evidence of unusual prion diseases. The majority of the cattle investigated were submitted to the VLA as BSE suspect during the years 1997-2005 and were reported to have clinical signs similar to BSE. We applied a combination of modified and previously unused diagnostic tests to this subgroup of cattle including lower concentrations of PK for protein digestion, tests that do not use PK for PrPsc detection and standard Western blot (WB) procedures with Mabs reactive with different regions of the PrP glycoprotein. A flow chart detailing the sequence for the investigation of potential unusual prion diseases of cattle are shown in Figure 1. 1) Determination of the lowest PK concentration that digests PrPc from brains of cattle The minimum concentration of PK required for the elimination of PrPc in the majority of non-exposed control cattle samples, resulting in a negative value in the Bio-Rad TeSeE ELISA, was determined. PK titrations were performed on BSE positive and negative control reference material (CRM) and subsequently on 47 individual confirmed negative brainstems. The brainstems had previously tested negative with the diagnostic Bio-Rad TeSeE ELISA by LGC and were obtained from active surveillance and therefore unlikely to have had clinical signs of disease. An amount of 0.3 µl/ml PK was selected for use in the adapted Bio-Rad TeSeE ELISA (0.3 Bio-Rad) (Table 1). 2) Determination of threshold values for the IDEXX HerdChek and 0.3 modified Bio-Rad rapid tests
The diagnostic tests have cut-off values that are set by the manufacturers. For the 0.3 Bio-Rad ELISA new cut-off values were determined to take account of the modifications. While no modifications were made to the IDEXX HerdChek assay cut-off values were calculated using the same test samples for consistency. 90 confirmed BSE negative brainstems were assayed and threshold values calculated as 3 standard deviations above the mean (Table 2). Threshold values of 0.166 Absorbance Units (AU) and 0.137 AU were set for the 0.3 Bio-Rad TeSeE and IDEXX Herdchek EIAs respectively. A single confirmed negative sample gave a value above the IDEXX threshold limit (0.240AU) on first assay. However, when repeated this sample was negative (0.016AU). 3) Results of assays applied to the test BSE cattle population
The assays described above and mapped in Figure 1 were then applied to the brains from 501 clinically suspect cattle. Following analysis the cattle were divided into five groups and these are described below. The results are summarised in Table 3. Group1: Confirmed negative diagnosis of clinically suspect cattle Brainstems from 501 cattle submitted to the VLA for BSE diagnosis between the years 1991 and 2005 that were subsequently diagnosed as negative by the tests used at time of slaughter, were assayed using the IDEXX and 0.3 Bio-Rad immunoassays for detection of abnormal PrP. 436 (87%) were negative by both tests. All of the samples submitted after 1999 were confirmed negative (see below) (Figure 8). By these criteria we were unable to detect abnormal PrP in the brainstems of these cattle and this subset of clinically suspect cattle is unlikely to harbour a prion disease. However, we were unable to test other areas of the brain from these cattle and PrPsc distribution patterns distinct from classical BSE cannot be ruled out. In addition to the 501 brainstems we also tested 191 cerebella by the same methods, all of which were negative by standard tests. Group 2: Confirmed positive for BSE by all diagnostic tests Sixty five samples remained that were positive in either the IDEXX or the 0.3 Bio-Rad assays or in both of these tests. Of these, 40 were positive by both tests (modified as above) and following retesting were positive using diagnostic concentrations of PK for the Bio-Rad TeSeE (figure 2). Immunohistochemical evaluation of abnormal PrP in the obex demonstrated normal distribution of PrPsc deposits similar to those observed for classical BSE (Figure 8).
To confirm that the PK resistant glycoproteins of abnormal PrP resembled the molecular profile of classical BSE, all 40 cases were immunoblotted using SHa31 MAb (figure 3). In all cases a signature 3 glycoprotein banding pattern was observed with relative mass and glycoprotein ratios indistinguishable from classical BSE. These animals ranged in age from 5 years to 12 years, with a mean age of 6 years, 10 months. All 40 animals were female and comprised 32 Friesians, 2 Holsteins, 2 Herefords, 1 Limousin/Friesian Cross, 2 Holstein/Friesian Cross and 1 Simmental.
The 40 confirmed positive samples were from cattle slaughtered between the years 1997 and 1999. We tested a total of 285 from this period and this represents 14.0% of the clinical suspects that were confirmed negative for BSE at this time. If this is representative of the entire clinical suspect unconfirmed cattle (total 2,426) during this period (1997-1999 inclusive) a total of 340 BSE positive cattle would have been missed. This under-diagnosis is likely to be a result of the diagnostic tests applied at the time. Up until the year 2000, all BSE cases were diagnosed by detection of vacuolation and gliosis in the obex. It is clear that this method is not 100% sensitive for prion diseases either because not all cases present with vacuolation or that vacuolation is a late onset phenomenon during clinical disease (Arnold et al., 2007). Our data showed that there were no additional cases of under-diagnosis after more sensitive diagnostic tests were introduced in 2000. During the years 1997-1999, a total of 12,171 clinical cases were submitted for BSE diagnosis of which 9,745 (80.1%) were confirmed positive with an estimated 2.8% of the total suspects submitted under-diagnosed by our calculations.
Assuming no other factors influenced the levels of correct diagnosis and that the numbers estimated for 1997 to 1999 were a true representation of the potential under-diagnosis of the entire epidemic up until 1999, then the total number of missed cases positive for BSE could have been in the region of 5,500.
A draft version of this manuscript has been prepared.
Group 3: Confirmed positive for BSE by all rapid diagnostic tests but negative by IHC
2 of the 501 negative subset brainstem tested were positive by standard biochemical, diagnostic tests (Table 4) but abnormal PrP deposits were not observed in the obex when evaluated by IHC (Figure 8). This is clearly an unusual finding and both cases were rigorously audited prior to further investigation to determine that the sample for biochemistry was identical to the paraffin-embedded sample. As far as can be determined no errors in sampling and dispatch occurred for these two samples. Further DNA profiling and matching frozen sample to histology processed sample would confirm this. There was insufficient sample to perform any further analysis on one case, but the other case was further investigated using the modified TeSeE Western blot protocol described above – at the diagnostic standard PK concentration of 4 µl/ml for PrPsc digestion. Western blotting of abnormal PrP from this sample confirmed the ELISA data with intense labelling of PK resistant PrP using the PrP-specific antibodies Sha31 and SAF84 (Figure 4a and 4b). The glycoprofile and molecular mass of the PrP bands were indistinguishable from classical BSE A band was labelled strongly with FH11 Mab (that recognises an N terminal PrP epitope) and is therefore likely to represent undigested PrP (Figure 4c). In addition, at 4 µl/ml PK, strong reactivity is also observed with the P4 mAb (Figure 4d). Molecular comparison of this case with classical BSE and with scrapie – using different levels of PK, different dilutions of positive sample and different PrP-specific antibodies, indicates that there is no discernible difference of the test sample with classical BSE. Both cases were extensively followed up by IHC using Mabs to different regions of the PrP molecule but were negative in all cases (data not shown).
Why the PrPsc could not be detected by IHC is unclear. Further analysis by transmission to rodent models of prion disease may shed further light on the characteristics of this sample. Indeed, murine models of prion disease have been reported where PrPsc cannot be detected in the brains but these studies confirmed the lack of PrPsc by all assays including Western Blot. Group 4: IDEXX Herdchek positive, 0.3 Bio-Rad negative, IHC positive. Two brainstem samples (98/00819; 98/02316) were positive by the IDEXX Herdchek EIA (Table 5) but Bio-Rad test negative even following PK digestion at sub-optimal concentrations. Both of these samples demonstrated abnormal PrP deposition in the obex by IHC evaluation (Figure 8). Western blot analysis of PK resistant PrP glycoprotein from sample 98/2316 indicated that low amounts of PrPres could be detected using Sha31 and SAF84 Mabs. From these blots and taking into account the low levels of PrPres detected the banding patterns appeared indistinguishable from classical BSE (Figure 5a and 5b). No further sample was available for 98/00819. The sample contained very low levels of PrPres as shown by the WB data and this is likely to be the reason for lack of signal in the Bio-Rad ELISA. At these levels of abnormal PrP we are at the threshold of detection. The IDEXX HerdChek assay has consistently shown a higher analytical sensitivity for classical scrapie in our hands than the Bio-Rad assays. The values for the IDEXX HerdChek were in the region of 0.15-0.88 and these values are much lower than any of the other samples we have tested in this study. These data suggest that the IDEXX assay is more analytically sensitive than the Bio-Rad TeSeE for BSE. However, there are alternative explanations for the discordance in test results. The Bio-Rad TeSeE ELISA detects PrPres with Mabs that detect 2 regions of the molecule. Any changes in PrP sequence in the region of Mab binding could alter analytical sensitivity. Therefore the bovine PrP open reading frame from 98/02316 was compared with that of two classical BSE samples, all three samples were 6:6 with respect to the octapeptide repeat. The only mutation seen in this unusual sample was at codon 78 and this is a “silent” mutation in that it does not affect the PrP protein sequence (glutamine, Q78). The Western blot results suggest that the PK cleavage sites of sample 98/02316 were not different from classical cases of BSE. Therefore we conclude that PrPres concentration in this sample was low, as indicated by the control BSE positive brain homogenate, when diluted to a level of 1/250, still producing bands of a far greater density than the test sample when assayed neat. Group 5: Diagnostic Bio-Rad and IDEXX negative, IHC negative but 0.3 Bio-Rad positive
Twenty-one of the clinical suspect brainstems tested by 0.3 Bio-Rad modified protocol had OD values above the calculated cut off point (range 0.166 to 0.857) (Figure 6) but were IDEXX Herdchek negative and IHC negative (figure 8). The samples were also diagnostic Bio-Rad TeSeE negative and the cattle, all female, ranged in age from 3 years to 11.5 years. They comprised Friesian, Friesian/Holsten, Hereford Cross, Aberdeen Angus Cross, Simmental Cross and Limousin Cross breeds. These samples, where sufficient tissue was available, were analysed, for the presence of partially PK resistant PrP, using the Bio-Rad Western blot protocol with digestion carried out at 20 and 0.3 µl/ml of PK and detected using the SHa31 Mab. Following digestion of the samples with 20 µl/ml PK the samples were shown to be negative for the characteristic PrPsc banding patterns when compared to three individual BSE-negative samples and a classical BSE positive sample (Fig 7a). However, faint bands were observed at approximately 16 and 25 KDa for 4 of the samples (T5, T8-T10) but this faint banding is consistent with partially digested PrPc but could also be a result of variable amounts of protein loaded per lane. At 0.3 µl/ml PK, banding is observed for all test samples, with banding consistent with partially digested PrPC, as also observed for the three known BSE-negative samples. In contrast, the classical BSE-positive sample gave a distinct banding pattern, different from that observed for the BSE-negative samples (Fig.7b). Consistent with the above results samples T5 and T8-T10 demonstrated increased intensity of labelling that could result from an up-regulation or increase in PrPc and could also account for the high signals in the modified ELISA.
Variable banding intensity between lanes may also be a result of inconsistent loading of amounts of protein per lane. However, our previous experience of testing protein concentrations PRIOR to PK digestion in the individual samples showed that they were very consistent to within <5% of the total amount. In addition, although we add pefabloc to stop PK digestion it is also likely that there is variation in the PK digestion amongst samples. Both variables could account for the differences in intensities between lanes. However, we cannot exclude the possibilitity that a PK sensitive variant of abnormal PrP is present as demonstrated by Barron et al 2007 who also demonstrated a 22 KDa band following sub optimal PK digestion. The samples were further investigated as below. Encephalitis may up regulate PrPc
One explanation for high values in the immunoassay following digestion with suboptimal concentrations of PK could be high levels of PrPc in the sample. Increased levels of PrPc may occur as a result of up-regulation of PrPc on tissue resident cells or from the influx of inflammatory cells into the site following infection. Differential diagnoses were available for 9 of the 21 animals and nine had confirmed encephalitic lesions and inflammation. Further to this observation we therefore analysed brainstems from 10 BSE negative cattle (but also clinical suspects) by both modified rapid tests that had confirmed encephalitis.
The brainstems from 9 encephalitis cattle were negative by both the 0.3 Bio-Rad TeSeE and IDEXX assays. The brainstem from 1 animal was positive by the 0.3 Bio-Rad assay but negative by the IDEXX EIA. The result from this sample is similar to the 21 observed above in group 5. It is unclear therefore whether the high levels of PrP are a result of concurrent infection as there is not a 100% correlation. However, PrPc is more susceptible to endogenous proteases and a low signal could be partly explained by inappropriate handling of the tissue at post-mortem. Loss of PrP detection following retesting of group 5 samples.
When all 21 samples were re-analysed from a fresh piece of tissue from the archive (likely to have been frozen and thawed by the archive staff) only one retested as positive (figure 6). Further analysis of this sample (sample number 99/00514) by Western blot has not shown any bands suggesting the presence of an atypical form of prion protein. Any PK sensitive PrP, whether PrPc or unusual prion disease-associated PrP, is likely to be affected by tissue handling techniques including freezing, thawing and the amount of time in storage. This could explain loss of signal. These samples may also represent a small number of outliers in the negative population. This is still higher than we would expect given that only 1/90 negative control samples were outliers in the original testing. Identification of Idiopathic Brainstem Neuronal Chromatolysis (IBNC) in group 5 samples One of the 21 samples identified in group 5 was shown to have IBNC following histological investigation (03/00002) (figure 8). Concurrently, we investigated the PrP distribution in known cases of IBNC (Jeffrey et al 2008; “Idiopathic Brainstem Neuronal Chromatolysis (IBNC): a novel prion protein related disorder of cattle?” BMC Vet Res. 2008 Sep 30;4:38. The IHC and histology profile of this case was very similar to that of the known IBNC cases. Investigation of the distribution and molecular characteristics of PrP from known IBNC See also: Idiopathic Brainstem Neuronal Chromatolysis (IBNC): a novel prion protein related disorder of cattle? Jeffrey M, Perez BB, Martin S, Terry L, González L. BMC Vet Res. 2008 Sep 30;4:38 Further investigations demonstrated that 57% the assays performed on the confirmed IBNC samples, using the 0.3 Bio-Rad TeSeE assay (n=42), gave values above those of the test kit control and also the BSE negative brain pool control. Half brains from six IBNC affected animals were retrieved from the TSE archive alongside the brainstem from a seventh animal. The cortex, brainstem, cerebellum and midbrain from these brains were sub-sampled and the adapted Bio-Rad TeSeE EIA, IDEXX Herdchek and Western Blot protocols applied to these tissues, in order to determine whether they could represent a form of atypical BSE. These samples had previously been found to be negative using the commercial Bio-Rad EIA and re-testing using this assay and the IDEXX Herdchek assay confirmed their negative status. When assayed using the adapted Bio-Rad protocol at 0.3µl/ml PK, 24/42 (57%) of the sample assays performed gave values above those of the test kit control and also the BSE negative brain pool control. Values above twice that of the calculated cut-off levels were found for each case but not for each brain site No PrPres was detected when Western blotting these samples at either 20 or 4µl/ml PK but a signal was detected on the gels when blotted at the 0.12 and 0.3µl/ml PK levels. At 0.12µl/ml PK the IBNC samples were indistinguishable from the negative controls but at the 0.3µl/ml level more PrPres was detected in the IBNC cases than in the controls with each of the antibodies tested (SHA31, F99, SAF84 and P4). Illustrations of the F99 blot are shown in the paper. Other data not shown.
These data suggest that IBNC affected cattle abnormally express or accumulate PrP in brain and that the abnormal PrP is not strongly resistant to protease digestion. The results suggest that either the range of prion diseases is still wider than previously thought or that abnormalities of prion protein expression may be associated with brain lesions unconnected with prion disorders. Biochemical and transmission studies are planned in order to investigate further (under SE2014). First case of H-type BSE identified in GB During the course of this study, 1/5 frozen brainstem from bovine BSE cases when immunoblotted using the Bio-Rad TeSeE Western blot with antibodies P4, L42, 6H4, Sha31 and SAF84, was found to have a PrP profile indistinguishable from French H-type BSE. This sample was the first case of H-type BSE to be identified in GB. It was a fallen 13-year-old Galloway cow, first tested and confirmed as a case of BSE in November 2005. Due to autolysis its brain was unsuitable for further characterisation by IHC. Its age and reported absence of clinical signs are consistent with other cases of H-type BSE.
When blotting the samples, mAbs Sha31 and 6H4 revealed, in this sample, an unglycosylated band with relative mobility less than BSE, and mAb P4, labelled the sample more strongly than the BSE samples hence supporting the observed similarities with the French H-type sample. Additionally, this study revealed in both this unusual sample and the French H-type a lower molecular weight band with relative mobility of between 6 and 10 kD labelled with the P4 and L42 mAbs. This band is not seen in BSE samples. This data was published in June 2007 (L. A. Terry et al. Veterinary record (2007) 160, 873-875). Discussion and Conclusions Here we report the investigation of 501 cattle samples that were submitted to the VLA for BSE diagnosis but subsequently confirmed as negative by the diagnostic test used at the time of submission. Prior to 2000 this was by histology alone and positive diagnosis was made solely upon the observation of vacuolation and gliosis in the relevant brain regions. As a result, using more sensitive diagnostic assays, we were able to diagnose BSE positive cattle from the years 1997-1999 inclusive that were originally negative by vacuolation. From these data we have estimated that approximately 3% of the total suspect cases submitted up until the year 1999 were mis-diagnosed. This is likely to be due to the relative sensitivities of the methods. In addition, it has been demonstrated in cattle that vacuolation occurs after PrPsc can be detected in the brain stem and that PrPsc is detected prior to clinical disease (Arnold et al, 2007). Thus these cattle may have suffering very early clinical signs. However, we have not ruled out the possibility that there may be a subset of BSE affected cattle where vacuolation at the obex does not occur. The two cattle that were positive by the rapid biochemical tests but negative by IHC is an unexplained observation. The samples both contained high amounts of abnormal prion protein as determined by the OD values from the rapid tests that according to our experience of confirmatory testing should have been easily detected by IHC. Furthermore, epitope mapping of the PK cleaved proteins demonstrated no unusual glycoform patterns and IHC evaluation with the same antibodies still did not reveal PrPd deposition in the wax embedded sections. Thus it is unlikely that lack of detection by IHC is the result of an unusual conformation of the PrPd that masks the epitope of R145, the antibody of choice for IHC evaluation at the VLA.
The two cattle that were positive by all tests except Bio-Rad ELISA are easier to explain. Previously we have demonstrated that the IDEXX HerdChek scrapie antigen EIA is more analytically sensitive for scrapie than the Bio-Rad ELISA (project SE2007) and this also appears to be the case for bovine BSE. Indeed the two samples were positive by the Bio-Rad Western blot but with significantly reduced signals compared to a bovine positive control. Samples in group 5 were only positive in the Bio-Rad ELISA and only if sub-optimal concentrations of PK were used. Several explanations could account for this result. First, the samples may contain a subset of PrP molecules that have a slightly higher resistance to PK digestion than normal PrPc and that it is not sufficiently aggregated to be detected by the IDEXX assay; whether this is related to a prion disease or some other event that confers such properties on normal PrP remains unanswered. There are notable descriptions in the literature of TSE models where disease is not accompanied by the characteristic accumulation of PK resistant PrP or was found at extremely low levels (Piccardo et al., 2007; Barron et al 2007; Nazor et al., 2005). These findings together might suggest an additional family of neurodegenerative diseases where the infectious form of PrP is not readily detected by our current diagnostic tests.
Second, the higher signal could be the result of an increase in the overall amount of PrPc in the samples as discussed in the results and related to up-regulation of PrP in cells resident in the brain or due to influx of inflammatory cells either as a result of damage or the presence of a non-prion related disease. Third, that the PrP in these samples is bound to an unidentified molecule that confers higher PK resistance, or fourth, inhibits proteinase K. IBNC is likely to represent a subset of this group of cattle. Based on these data, our overall conclusion is that a second type of BSE is unlikely to have co-existed at a high prevalence with the classical form in the cattle population during the UK epidemic.
Final Report - Annex : Atypical prion proteins in cattle (10064k)
Final Report - SID5 : Atypical prion proteins in cattle (201k)
USDA announces expanded BSE surveillance program Filed Under: BSE
By: Marty Heiberg | Mar 15, 2004 Editor's note: Some material was added to this story Mar 16.
Mar 15, 2004 (CIDRAP News) – Secretary of the US Department of Agriculture (USDA) Ann Veneman this afternoon announced an expanded program of surveillance for bovine spongiform encephalopathy (BSE) in the United States. Preparations for the increased testing will begin immediately and the program is expected to be fully operational by June 1. The new testing procedures will be in place for 12 to 18 months, after which an assessment will determine future plans.
"The intensive one-time surveillance effort will allow us to determine more accurately whether BSE is present in the US cattle population, and if so, estimate the level of disease. By expanding our surveillance, we will be able to provide consumers, trading partners, and industry increased assurances about the BSE status of the U.S. cattle population," states the new plan, which was published on the USDA Web site today.
The new plan incorporates last month's recommendations from the international scientific review panel and it is supported by the Harvard Center for Risk Analysis, Veneman said at a press briefing. It calls for testing a much larger number of specimens from the high-risk BSE cattle population than the current 40,000 per year as well as about 20,000 random samples from normal-appearing adult cows.
Cattle at high risk for BSE are estimated to number approximately 446,000 currently in the United States. The definition of high risk, based on experience in the United Kingdom and Europe, includes adult cattle that are nonambulatory ("downers"), dead on the farm, or showing clinical signs consistent with BSE.
Ron DeHaven, the USDA's chief veterinary officer, said at the briefing that the expanded program would mean testing "as many as we possibly can" of the target population of cattle. He explained that the new testing would allow for identification of BSE at a rate of 1 in 10 million cattle with a confidence level of 95% if 201,000 samples were tested and a confidence level of 99% if 268,000 samples were tested.
Testing will be done at 17 state and university laboratories, with confirmation of any positive results at the National Veterinary Services Laboratory in Ames, Iowa. Funding for the new program totals $70 million.
When questioned about proposals to test 100% of cattle, DeHaven said that science does not justify this level of testing and that, while the USDA is still evaluating the proposals, testing at this level would be solely for marketability and export purposes. The USDA's newly enhanced program, he said, is strictly for surveillance purposes and will determine whether and at what level BSE exists in the target cattle population.
DeHaven said the expanded testing program will rely on rapid screening tests, several of which the USDA is currently evaluating. "We would anticipate in two or three months' time being able to license perhaps a couple or more of those tests," he said.
Because the screening tests are designed to be very sensitive, some false-positive results are expected, DeHaven said, adding, "That's just the nature of the beast." The national laboratory in Ames will use immunohistochemical staining, considered the "gold standard" in BSE testing, to confirm any positives.
DeHaven said the USDA has made no decision yet on the proposal by Creekstone Farms of Arkansas City, Kan., to test all its cattle so the beef can be exported to Japan and other Asian markets.
The USDA will collect samples from high-risk cattle at a variety of places, including federally inspected slaughter plants, farms, rendering plants, veterinary diagnostic laboratories, pet food plants, and livestock sale barns, DeHaven said.
To test a random sample of healthy older cattle, the USDA will focus its main efforts on 40 slaughter plants in 17 states, according to DeHaven. Those plants slaughter more than 86% of all cattle in the nation, he said.
Under questioning, DeHaven refused to give a specific target for the number of high-risk cattle to be tested. "To estimate how many we will be able to collect is simply premature," he said. "It's possible that we would collect somewhere less than 200,000 and still have a very statistically valid sampling."
DeHaven said USDA veterinarians will work with state veterinarians and other state officials to develop plans for collecting cattle samples for testing in each state.
See also:
Transcript of USDA's Mar 15 news briefing http://www.usda.gov/Newsroom/0106.04.html
Robert Roos, CIDRAP News Editor, contributed to this story.
USDA did not test possible mad cows
By Steve Mitchell
United Press International
Published 6/8/2004 9:30 PM
WASHINGTON, June 8 (UPI) -- The U.S. Department of Agriculture claims it tested 500 cows with signs of a brain disorder for mad cow disease last year, but agency documents obtained by United Press International show the agency tested only half that number.
i almost forgot LOL;
BESIDES THE TEXAS MAD COW THAT WAS RENDERED AND NEVER TESTED;
On Friday, April 30 th , the Food and Drug Administration learned that a cow with central nervous system symptoms had been killed and shipped to a processor for rendering into animal protein for use in animal feed.
FDA, which is responsible for the safety of animal feed, immediately began an investigation. On Friday and throughout the weekend, FDA investigators inspected the slaughterhouse, the rendering facility, the farm where the animal came from, and the processor that initially received the cow from the slaughterhouse.
FDA's investigation showed that the animal in question had already been rendered into "meat and bone meal" (a type of protein animal feed). Over the weekend FDA was able to track down all the implicated material. That material is being held by the firm, which is cooperating fully with FDA. ...
http://www.fda.gov/bbs/topics/news/2004/NEW01061.html
USDA orders silence on mad cow in Texas
By Steve Mitchell United Press International Published 5/11/2004 10:16 PM
WASHINGTON, May 11 (UPI) -- The U.S. Department of Agriculture has issued an order instructing its inspectors in Texas, where federal madcow disease testing policies recently were violated, not to talk about the cattle disorder with outside parties, United Press International has learned.
The order, sent May 6 by e-mail from the USDA's Dallas district office,was issued in the wake of the April 27 case at Lone Star Beef in San Angelo, in which a cow displaying signs of a brain disorder was not tested for mad cow disease despite a federal policy to screen all such animals.
The deadly illness also is known as bovine spongiform encephalopathy.
Both the USDA and its Inspector General -- amid allegations that an offsite supervisor overruled the opinion of the inspectors on site and made the final decision not to test the animal -- have opened up investigations to determine why agency policy was violated.
The order, which was obtained by UPI, was issued by Ijaz Qazi, circuit supervisor for the USDA's Food Safety and Inspection Service's Dallas district, which covers the entire state. It reads: "All BSE inquiries MUST be directed to Congressional Public Affairs Phone 202-720-9113 attention Rob Larew OR Steve Khon. This is an urgent message. Any question contact me. Ijaz Qazi."
Although the language might sound innocuous, experienced inspectors familiar with USDA parlance have taken to referring to the notice as a "gag order."
The National Joint Council of Food Inspection Locals -- the national inspectors union -- considers the order a violation of inspectors' freespeech rights and is considering legal action against the USDA for breaching the labor agreement they have with the agency.
Inspectors alleged the order also suggests the agency is concerned about its personnel leaking damaging information about either the Texas case or the USDA's overall mad cow disease surveillance program, which has come under fire since the discovery of an infected cow in Washington state last December.
"Anytime the government suppresses an individual's freedom of speech,that's unconstitutional," Gary Dahl, president of Local 925, the Colorado inspectors union, told UPI.
Stanley Painter, chairman of the National Joint Council, said the USDA has sent out notices in the past stating inspectors cannot talk to reporters.
"It's an intimidation thing," Painter told UPI. Inspectors have the right to talk to anybody about any subject, as long as they clarify they are not speaking on behalf of the USDA and they are not doing it on government time, he said.
USDA spokesman Steven Cohen said he was not familiar with the notice from the Dallas office. He said he would look into it, but did not respond by UPI's publication time. In general, Cohen said, "There's an expectation any statement on behalf of the agency would come from the office of communications (in Washington.)"
Asked if employees could speak freely as long as they clarified that their views did not reflect those of the agency, Cohen said, "We'd rather that agency policy be communicated by those in a position to speak for the agency."
Qazi told UPI the notice was not issued in conjunction with the Texas case and it was routine agency practice that outside inquiries be referred to the Washington office. He said inspectors are free to talk to outside parties, including reporters, and he did not consider the e-mail a violation of the labor agreement with the inspectors.
Painter said the USDA's efforts to keep its employees from talking about mad cow would be better spent "with issues like protecting the consuming public instead of trying to hide things." He added he would "just about bet his last nickel" agency management was attempting to suppress information about the Texas case.
"To keep federal employees from reporting government waste, misuse ofappropriations -- those types of things -- that's not a good thing either," Dahl said. "If there is something wrong, let's get it out in the open -- let's get it fixed. We're working for the public, the American consumers. I think they have the right to know this," he said.
"And believe me there's so many indicators saying that the USDA's madcow testing program is broken," Dahl added.
At least one member of Congress, Sen. Tom Harkin, D-Iowa, agrees.
Harkin, a long-time critic of the USDA, sent a letter to Agriculture Secretary Ann Veneman on Monday, saying the Texas incident "calls into question the effectiveness and reliability of USDA's current and proposed surveillance system."
The USDA has proposed testing more than 200,000 cows -- or 10 times its current rate -- in an expanded program scheduled to begin June 1. Harkin wrote in the five-page letter, however, that given the realities of the cattle industry, it is "quite doubtful" the USDA will be able to test that many cows, particularly because it had difficulty finding 20,000 last year.
"We simply cannot tolerate a BSE testing system that fails to give valid answers to critical questions for U.S. consumers and foreign customers,"Harkin said in the letter, which sharply criticizes the agency's failure to address explicitly how its new surveillance program will be implemented.
"We look forward to receiving (Harkin's) letter and having the opportunity to review it and respond to him," USDA spokesman Ed Loyd told UPI. "USDA has acknowledged there was a failure in not testing that cow in Texas for BSE, so we are all working to ensure that does not occur again."
Jim Rogers, a spokesman for USDA's Animal and Plant Health InspectionService, which oversees the agency's mad cow surveillance program, told UPI the agency has tested about 15,500 animals since fiscal year 2004 began, on Oct. 1, 2003. However, the agency has refused to identify the states and facilities from which the cows originated. Rogers said UPI would have to seek that information through the Freedom of Information Act.
The question is central to the USDA's implementation of its expanded surveillance program. Downer cows -- those unable to stand or walk --made up the bulk of the animals the agency tested for mad cow inprevious years, but these were banned from being slaughtered for human consumption in December. This means the agency inspectors no longer can obtain brain samples from these cows at slaughterhouses as they could in the past.
Furthermore, the USDA has not provided any evidence it has worked out agreements with rendering facilities or ranchers, where downers and dead cows are now most likely to be found, to obtain the extra animals for testing.
Loyd said the agency is "working very hard to get animals on the farm that would never show up in a processing facility," and he was "not aware of any issues" that would delay the launch of the new program.
However, he was unable to provide the names or locations of the rendering facilities where the agency will be obtaining cow brains for BSE testing. He said he would look into it but did not return two follow-up phone calls from UPI before publication.
--
Steve Mitchell is UPI's Medical Correspondent. E-mail sciencemail@upi.com
Copyright © 2001-2004 United Press International
THE REST IS HISTORY, more atypical bse mad cow cases were showing up, testing questionable to say the least, i remind you of the infamous BSE ENHANCED and SUPRESSED BSE SURVEILLANCE AND THE HARVARD BSE BS that followed, and why the infamous ENHANCED BSE SURVEILLANCE AND TESTING WAS SHUT DOWN...terry
Audit Report Animal and Plant Health Inspection Service Bovine Spongiform Encephalopathy (BSE) Surveillance Program – Phase II
and
Food Safety and Inspection Service Controls Over BSE Sampling, Specified Risk Materials, and Advanced Meat Recovery Products - Phase III
"These 9,200 cases were different because brain tissue samples were preserved with formalin, which makes them suitable for only one type of test--immunohistochemistry, or IHC."
THIS WAS DONE FOR A REASON!
THE IHC test has been proven to be the LEAST LIKELY to detect BSE/TSE in the bovine, and these were probably from the most high risk cattle pool, the ones the USDA et al, SHOULD have been testing. ...TSS
TEXAS 2ND MAD COW THAT WAS COVERED UP, AFTER AN ACT OF CONGRESS, AND CALLS FROM TSE PRION SCIENTIST AROUND THE GLOBE, THIS 2ND MAD COW IN TEXAS WAS CONFIRMED
THE USDA MAD COW FOLLIES POSITIVE TEST COVER UP
JOHANNS SECRET POSTIVE MAD COW TEST THAT WERE IGNORED
OIG AND THE HONORABLE FONG CONFIRMS TEXAS MAD AFTER AN ACT OF CONGRESS 7 MONTHS LATER
TEXAS MAD COW
THEY DID FINALLY TEST AFTER SITTING 7+ MONTHS ON A SHELF WHILE GW BORE THE BSE MRR POLICY, i.e. legal trading of all strains of TSE. now understand, i confirmed this case 7 months earlier to the TAHC, and then, only after i contacted the Honorable Phyllis Fong and after an act of Congress, this animal was finally confirmed ;
During the course of the investigation, USDA removed and tested a total of 67 animals of interest from the farm where the index animal's herd originated. All of these animals tested negative for BSE. 200 adult animals of interest were determined to have left the index farm. Of these 200, APHIS officials determined that 143 had gone to slaughter, two were found alive (one was determined not to be of interest because of its age and the other tested negative), 34 are presumed dead, one is known dead and 20 have been classified as untraceable. In addition to the adult animals, APHIS was looking for two calves born to the index animal. Due to record keeping and identification issues, APHIS had to trace 213 calves. Of these 213 calves, 208 entered feeding and slaughter channels, four are presumed to have entered feeding and slaughter channels and one calf was untraceable.
see new link;
Executive Summary In June 2005, an inconclusive bovine spongiform encephalopathy (BSE) sample from November 2004, that had originally been classified as negative on the immunohistochemistry test, was confirmed positive on SAF immunoblot (Western blot). The U.S. Department of Agriculture (USDA) identified the herd of origin for the index cow in Texas; that identification was confirmed by DNA analysis. USDA, in close cooperation with the Texas Animal Health Commission (TAHC), established an incident command post (ICP) and began response activities according to USDA’s BSE Response Plan of September 2004. Response personnel removed at-risk cattle and cattle of interest (COI) from the index herd, euthanized them, and tested them for BSE; all were negative. USDA and the State extensively traced all at-risk cattle and COI that left the index herd. The majority of these animals entered rendering and/or slaughter channels well before the investigation began. USDA’s response to the Texas finding was thorough and effective.
snip...
Trace Herd 3 The owner of Trace Herd 3 was identified as possibly having received an animal of interest. The herd was placed under hold order on 7/27/05. The herd inventory was conducted on 7/28/05. The animal of interest was not present within the herd, and the hold order was released on 7/28/05. The person who thought he sold the animal to the owner of Trace Herd 3 had no records and could not remember who else he might have sold the cow to. Additionally, a search of GDB for all cattle sold through the markets by that individual did not result in a match to the animal of interest. The animal of interest traced to this herd was classified as untraceable because all leads were exhausted.
Trace Herd 4 The owner of Trace Herd 4 was identified as having received one of the COI through an order buyer. Trace Herd 4 was placed under hold order on 7/29/05. A complete herd inventory was conducted on 8/22/05 and 8/23/05. There were 233 head of cattle that were examined individually by both State and Federal personnel for all man-made identification and brands. The animal of interest was not present within the herd. Several animals were reported to have died in the herd sometime after they arrived on the premises in April 2005. A final search of GDB records yielded no further results on the eartag of interest at either subsequent market sale or slaughter. With all leads having been exhausted, this animal of interest has been classified as untraceable. The hold order on Trace Herd 4 was released on 8/23/05.
Trace Herd 5 The owner of Trace Herd 5 was identified as having received two COI and was placed under hold order on 8/1/05. Trace Herd 5 is made up of 67 head of cattle in multiple pastures. During the course of the herd inventory, the owner located records that indicated that one of the COI, a known birth cohort, had been sold to Trace Herd 8 where she was subsequently found alive. Upon completion of the herd inventory, the other animal of interest was not found within the herd. A GDB search of all recorded herd tests conducted on Trace Herd 5 and all market sales by the owner failed to locate the identification tag of the animal of interest and she was subsequently classified as untraceable due to all leads having been exhausted. The hold order on Trace Herd 5 was released on 8/8/05.
Trace Herd 6 The owner of Trace Herd 6 was identified as possibly having received an animal of interest and was placed under hold order on 8/1/05. This herd is made up of 58 head of cattle on two pastures. A herd inventory was conducted and the animal of interest was not present within the herd. The owner of Trace Herd 6 had very limited records and was unable to provide further information on where the cow might have gone after he purchased her from the livestock market. A search of GDB for all cattle sold through the markets by that individual did not result in a match to the animal of interest. Additionally, many of the animals presented for sale by the owner of the herd had been re-tagged at the market effectually losing the traceability of the history of that animal prior to re-tagging. The animal of interest traced to this herd was classified as untraceable due to all leads having been exhausted. The hold order on Trace Herd 6 was released on 8/3/05.
Trace Herd 7 The owner of Trace Herd 7 was identified as having received an animal of interest and was placed under hold order on 8/1/05. Trace Herd 7 contains 487 head of cattle on multiple pastures in multiple parts of the State, including a unit kept on an island. The island location is a particularly rough place to keep cattle and the owner claimed to have lost 22 head on the island in 2004 due to liver flukes. Upon completion of the herd inventory, the animal of interest was not found present within Trace Herd 7. A GDB search of all recorded herd tests conducted on Trace Herd 7 and all market sales by the owner failed to locate the identification tag of the animal of interest. The cow was subsequently classified as untraceable. It is quite possible though that she may have died within the herd, especially if she belonged to the island unit. The hold order on Trace Herd 7 was released on 8/8/05.
Owner and Corporation Plead Guilty to Defrauding Bovine Spongiform Encephalopathy (BSE) Surveillance Program
An Arizona meat processing company and its owner pled guilty in February 2007 to charges of theft of Government funds, mail fraud, and wire fraud. The owner and his company defrauded the BSE Surveillance Program when they falsified BSE Surveillance Data Collection Forms and then submitted payment requests to USDA for the services. In addition to the targeted sample population (those cattle that were more than 30 months old or had other risk factors for BSE), the owner submitted to USDA, or caused to be submitted, BSE obex (brain stem) samples from healthy USDA-inspected cattle. As a result, the owner fraudulently received approximately $390,000. Sentencing is scheduled for May 2007.
snip...
4 USDA OIG SEMIANNUAL REPORT TO CONGRESS FY 2007 1st Half
Audit Report Animal and Plant Health Inspection Service Bovine Spongiform Encephalopathy (BSE) Surveillance Program – Phase II and Food Safety and Inspection Service Controls Over BSE Sampling, Specified Risk Materials, and Advanced Meat Recovery Products - Phase III
UNITED STATES DEPARTMENT OF AGRICULTURE OFFICE OF INSPECTOR GENERAL Washington, D.C. 20250 January 25, 2006 REPLY TO ATTN OF: 50601-10-KC TO: W. Ron DeHaven Administrator Animal and Plant Health Inspection Service Barbara Masters Administrator Food Safety and Inspection Service
ATTN: William J. Hudnall Deputy Administrator Marketing Regulatory Program Business Services William C. Smith Assistant Administrator Office of Program Evaluation, Enforcement, and Review FROM: Robert W. Young /s/ Assistant Inspector General for Audit
SUBJECT: Animal and Plant Health Inspection Service - Bovine Spongiform Encephalopathy (BSE) Surveillance Program - Phase II and Food Safety and Inspection Service - Controls Over BSE Sampling, Specified Risk Materials, and Advanced Meat Recovery Products - Phase III
This report presents the results of our audit of the enhanced BSE surveillance program and controls over specified risk materials and advanced meat recovery products. Your written response to the official draft report, dated January 20, 2006, is included as exhibit G with excerpts of the response and the Office of Inspector General’s (OIG) position incorporated into the Findings and Recommendations section of the report, where applicable. We accept the management decisions for all recommendations. Please follow your agency’s internal procedures in forwarding documentation for final action to the Office of the Chief Financial Officer (OCFO). We are providing a separate memorandum to the agencies and OCFO that provides specific information on the actions to be completed to achieve final action. We appreciate your timely response and the cooperation and assistance provided to our staff during the audit USDA/OIG-A/50601-10-KC/ Page i
Executive Summary
Animal and Plant Health Inspection Service - Bovine Spongiform Encephalopathy (BSE) Surveillance Program - Phase II and Food Safety and Inspection Service - Controls Over BSE Sampling, Specified Risk Materials, and Advanced Meat Recovery Products - Phase III
Results in Brief This report evaluates elements of the interlocking safeguards in place to protect United States (U.S.) beef from Bovine Spongiform Encephalopathy, widely known as BSE or "mad cow disease." Since 1990, the U.S. Department of Agriculture (USDA), Animal and Plant Health Inspection Service (APHIS), has led a multi-agency effort to monitor and prevent BSE from entering the food supply. After discovering a BSE-positive cow in December 2003, APHIS expanded its BSE surveillance program. To further protect the food supply, USDA banned materials identified as being at risk of carrying BSE (specified risk materials (SRM)), such as central nervous system tissue. As part of this effort, USDA’s Food Safety and Inspection Service (FSIS) required beef slaughter and processing facilities to incorporate controls for handling such materials into their operational plans. Onsite FSIS inspectors also inspect cattle for clinical signs in order to prevent diseased animals from being slaughtered for human consumption. To evaluate the effectiveness of the safeguards, we assessed APHIS’ implementation of the expanded surveillance program, as well as FSIS’ controls to prevent banned SRMs from entering the food supply.
In June 2004, APHIS implemented its expanded surveillance program; participation by industry in this surveillance program is voluntary. As of May 2005, over 350,000 animals were sampled and tested for BSE. To date, two animals tested positive for BSE; one tested positive after implementation of the expanded surveillance program.
USDA made significant efforts to implement the expanded BSE surveillance program. Much needed to be done in a short period of time to establish the necessary processes, controls, infrastructure, and networks to assist in this effort. In addition, extensive outreach and coordination was undertaken with other Federal, State, and local entities, private industry, and laboratory and veterinary networks. This report provides an assessment as to the progress USDA made in expanding its surveillance effort and the effectiveness of its controls and processes. This report also discusses the limitations of its program and data in assessing the prevalence of BSE in the U.S. herd.
snip...
40 ELISA test procedures require two additional (duplicate) tests if the initial test is reactive, before final interpretation. If either of the duplicate tests is reactive, the test is deemed inconclusive.
41 Protocol for BSE Contract Laboratories to Receive and Test Bovine Brain Samples and Report Results for BSE Surveillance Standard Operating Procedure (SOP), dated October 26, 2004.
42 The NVSL conducted an ELISA test on the original material tested at the contract laboratory and on two new cuts from the sample tissue.
43 A visual examination of brain tissue by a microscope.
44 A localized pathological change in a bodily organ or tissue.
SNIP...
PLEASE SEE FLAMING EVIDENCE THAT THE USDA ET AL COVERED UP MAD COW DISEASE IN TEXAS ;
PAGE 43;
Section 2. Testing Protocols and Quality Assurance Controls
snip...
FULL TEXT 130 PAGES
Comments on technical aspects of the risk assessment were then submitted to FSIS.
Comments were received from Food and Water Watch, Food Animal Concerns Trust (FACT), Farm Sanctuary, R-CALF USA, Linda A Detwiler, and Terry S. Singeltary.
This document provides itemized replies to the public comments received on the 2005 updated Harvard BSE risk assessment. Please bear the following points in mind:
Owens, Julie From: Terry S. Singeltary Sr. [flounder9@verizon.net]
Sent: Monday, July 24, 2006 1:09 PM To: FSIS Regulations Comments
Subject: [Docket No. FSIS-2006-0011] FSIS Harvard Risk Assessment of Bovine Spongiform Encephalopathy (BSE) Page 1 of 98 8/3/2006
Greetings FSIS, I would kindly like to comment on the following ;
Suppressed peer review of Harvard study October 31, 2002.
October 31, 2002 Review of the Evaluation of the Potential for Bovine Spongiform Encephalopathy in the United States Conducted by the Harvard Center for Risk Analysis, Harvard School of Public Health and Center for Computational Epidemiology, College of Veterinary Medicine, Tuskegee University Final Report Prepared for U.S. Department of Agriculture Food Safety and Inspection Service Office of Public Health and Science Prepared by RTI Health, Social, and Economics Research Research Triangle Park, NC 27709 RTI Project Number 07182.024
FULL TEXT OF GOA REPORT BELOW (takes a while to load)
2. Mad Cow Disease: Improvements in the Animal Feed Ban and Other Regulatory Areas Would Strengthen U.S. Prevention Efforts. GAO-02-183, January 25.
SATURDAY, AUGUST 16, 2008
Qualitative Analysis of BSE Risk Factors in the United States February 13, 2000 at 3:37 pm PST (BSE red book)
Tuesday, September 14, 2010
Transmissible Spongiform Encephalopathies Advisory Committee; Notice of Meeting October 28 and 29, 2010 (COMMENT SUBMISSION)
FULL TEXT OF GOA REPORT BELOW (takes a while to load)
2. Mad Cow Disease: Improvements in the Animal Feed Ban and Other Regulatory Areas Would Strengthen U.S. Prevention Efforts. GAO-02-183, January 25.
8 hr BSE confirmation turnaround took 7+ months to confirm this case, so the BSE MRR policy could be put into place. ...TSS
-------- Original Message --------
Subject: re-USDA's surveillance plan for BSE aka mad cow disease
Date: Mon, 02 May 2005 16:59:07 -0500
From: "Terry S. Singeltary Sr."
Greetings Honorable Paul Feeney, Keith Arnold, and William Busbyet al at OIG, ...............
snip...
There will be several more emails of my research to follow. I respectfully request a full inquiry into the cover-up of TSEs in the United States of America over the past 30 years. I would be happy to testify...
Thank you, I am sincerely, Terry S. Singeltary Sr. P.O. Box , Bacliff, Texas USA 77518 xxx xxx xxxx
Date: June 14, 2005 at 1:46 pm PST
In Reply to:
Re: Transcript Ag. Secretary Mike Johanns and Dr. John Clifford, Regarding further analysis of BSE Inconclusive Test Results
posted by TSS on June 13, 2005 at 7:33 pm:
Secretary of Agriculture Ann M. Veneman resigns Nov 15 2004, three days later inclusive Mad Cow is announced. June 7th 2005 Bill Hawks Under Secretary for Marketing and Regulatory Programs resigns. Three days later same mad cow found in November turns out to be positive. Both resignation are unexpected. just pondering... TSS
*** 2009 UPDATE ON ALABAMA AND TEXAS MAD COWS 2005 and 2006 ***
Suppressed peer review of Harvard study October 31, 2002
October 31, 2002
Review of the Evaluation of the Potential for Bovine Spongiform Encephalopathy in the United States Conducted by the Harvard Center for Risk Analysis, Harvard School of Public Health and Center for Computational Epidemiology, College of Veterinary Medicine, Tuskegee University
Final Report
Harvard Risk Assessment of Bovine Spongiform Encephalopathy (BSE) Update; Notice of Availability and Technical Meeting
Owens, Julie
From: Terry S. Singeltary Sr. [flounder9@verizon.net]
Sent: Monday, July 24, 2006 1:09 PM
To: FSIS RegulationsComments
Subject: [Docket No. FSIS-2006-0011] FSIS Harvard Risk Assessment of Bovine Spongiform Encephalopathy (BSE)
Response to Public Comments on the Harvard Risk Assessment of Bovine Spongiform Encephalopathy Update,
October 31, 2005
INTRODUCTION
The United States Department of Agriculture’s Food Safety and Inspection Service (FSIS) held a public meeting on July 25, 2006 in Washington, D.C. to present findings from the Harvard Risk Assessment of Bovine Spongiform Encephalopathy Update, October 31, 2005 (report and model located on the FSIS website: http://www.fsis.usda.gov/Science/Risk_Assessments/index.asp). Comments on technical aspects of the risk assessment were then submitted to FSIS. Comments were received from Food and Water Watch, Food Animal Concerns Trust (FACT), Farm Sanctuary, RCALF USA, Linda A Detwiler, and Terry S. Singeltary. This document provides itemized replies to the public comments received on the 2005 updated Harvard BSE risk assessment. Please bear the following points in mind:
03-025IFA
03-025IFA-2
Terry S. Singeltary
From: Terry S. Singeltary Sr. [flounder9@verizon.net]
Sent: Thursday, September 08, 2005 6:17 PM
To: fsis.regulationscomments@fsis.usda.gov
Subject: [Docket No. 03-025IFA] FSIS Prohibition of the Use of Specified Risk Materials for Human Food and Requirements for the Disposition of Non-Ambulatory Disabled Cattle
ONE final comment tonight, i just cannot take anymore. well, ill just let the facts speak for themselves, no need to even comment ;
Section 2. Testing Protocols and Quality Assurance Controls
In November 2004, USDA announced that its rapid screening test, Bio-Rad Enzyme Linked Immunosorbent Assay (ELISA), produced an inconclusive BSE test result as part of its enhanced BSE surveillance program. The ELISA rapid screening test performed at a BSE contract laboratory produced three high positive reactive results.40 As required,41 the contract laboratory forwarded the inconclusive sample to the APHIS National Veterinary Services Laboratories (NVSL) for confirmatory testing. NVSL repeated the ELISA testing and again produced three high positive reactive results.42 In accordance with its established protocol, NVSL ran its confirmatory test, an immunohistochemistry (IHC) test, which was interpreted as negative for BSE. In addition, NVSL performed a histological43 examination of the tissue and did not detect lesions44 consistent with BSE.
Faced with conflicting results, NVSL scientists recommended additional testing to resolve the discrepancy but APHIS headquarters officials concluded no further testing was necessary because testing protocols were followed. In our discussions with APHIS officials, they justified their decision not to do additional testing because the IHC is internationally recognized as the "gold standard." Also, they believed that conducting additional tests would undermine confidence in USDA’s established testing protocols.
full text 130 pages ;
PDF]Freas, William TSS SUBMISSION
File Format: PDF/Adobe Acrobat - Page 1. J Freas, William From: Sent: To: Subject: Terry S. Singeltary Sr. [flounder@wt.net] Monday, January 08,200l 3:03 PM freas ... http://web.archive.org/web/20170301223601/https://www.fda.gov/OHRMS/DOCKETS/AC/01/slides/3681s2_09.pdf
Sunday, January 10, 2021APHIS Concurrence With OIE Risk Designation for Bovine Spongiform Encephalopathy [Docket No. APHIS-2018-0087] Singeltary Submission June 17, 2019APHIS Concurrence With OIE Risk Designation for Bovine Spongiform Encephalopathy [Docket No. APHIS-2018-0087] Singeltary SubmissionGreetings APHIS et al,I would kindly like to comment on APHIS Concurrence With OIE Risk Designation for Bovine Spongiform Encephalopathy [Docket No. APHIS-2018-0087], and my comments are as follows, with the latest peer review and transmission studies as references of evidence.THE OIE/USDA BSE Minimal Risk Region MRR is nothing more than free pass to import and export the Transmissible Spongiform Encephalopathy TSE Prion disease. December 2003, when the USDA et al lost it's supposedly 'GOLD CARD' ie BSE FREE STATUS (that was based on nothing more than not looking and not finding BSE), once the USA lost it's gold card BSE Free status, the USDA OIE et al worked hard and fast to change the BSE Geographical Risk Statuses i.e. the BSE GBR's, and replaced it with the BSE MRR policy, the legal tool to trade mad cow type disease TSE Prion Globally. The USA is doing just what the UK did, when they shipped mad cow disease around the world, except with the BSE MRR policy, it's now legal.Also, the whole concept of the BSE MRR policy is based on a false pretense, that atypical BSE is not transmissible, and that only typical c-BSE is transmissible via feed. This notion that atypical BSE TSE Prion is an old age cow disease that is not infectious is absolutely false, there is NO science to show this, and on the contrary, we now know that atypical BSE will transmit by ORAL ROUTES, but even much more concerning now, recent science has shown that Chronic Wasting Disease CWD TSE Prion in deer and elk which is rampant with no stopping is sight in the USA, and Scrapie TSE Prion in sheep and goat, will transmit to PIGS by oral routes, this is our worst nightmare, showing even more risk factors for the USA FDA PART 589 TSE PRION FEED ban.The FDA PART 589 TSE PRION FEED ban has failed terribly bad, and is still failing, since August 1997. there is tonnage and tonnage of banned potential mad cow feed that went into commerce, and still is, with one decade, 10 YEARS, post August 1997 FDA PART 589 TSE PRION FEED ban, 2007, with 10,000,000 POUNDS, with REASON, Products manufactured from bulk feed containing blood meal that was cross contaminated with prohibited meat and bone meal and the labeling did not bear cautionary BSE statement. you can see all these feed ban warning letters and tonnage of mad cow feed in commerce, year after year, that is not accessible on the internet anymore like it use to be, you can see history of the FDA failure August 1997 FDA PART 589 TSE PRION FEED ban here, but remember this, we have a new outbreak of TSE Prion disease in a new livestock species, the camel, and this too is very worrisome.WITH the OIE and the USDA et al weakening the global TSE prion surveillance, by not classifying the atypical Scrapie as TSE Prion disease, and the notion that they want to do the same thing with typical scrapie and atypical BSE, it's just not scientific.WE MUST abolish the BSE MRR policy, go back to the BSE GBR risk assessments by country, and enhance them to include all strains of TSE Prion disease in all species. With Chronic Wasting CWD TSE Prion disease spreading in Europe, now including, Norway, Finland, Sweden, also in Korea, Canada and the USA, and the TSE Prion in Camels, the fact the the USA is feeding potentially CWD, Scrapie, BSE, typical and atypical, to other animals, and shipping both this feed and or live animals or even grains around the globe, potentially exposed or infected with the TSE Prion. this APHIS Concurrence With OIE Risk Designation for Bovine Spongiform Encephalopathy [Docket No. APHIS-2018-0087], under it's present definition, does NOT show the true risk of the TSE Prion in any country. as i said, it's nothing more than a legal tool to trade the TSE Prion around the globe, nothing but ink on paper.AS long as the BSE MRR policy stays in effect, TSE Prion disease will continued to be bought and sold as food for both humans and animals around the globe, and the future ramifications from friendly fire there from, i.e. iatrogenic exposure and transmission there from from all of the above, should not be underestimated. ...
APHIS Indemnity Regulations [Docket No. APHIS-2021-0010] RIN 0579-AE65 Singeltary Comment SubmissionComment from Singeltary Sr., TerryPosted by the Animal and Plant Health Inspection Service on Sep 8, 2022SO, WHO'S UP FOR SOME MORE TSE PRION POKER, WHO'S ALL IN $$$
SO, ATYPICAL SCRAPIE ROUGHLY HAS 50 50 CHANCE ATYPICAL SCRAPIE IS CONTAGIOUS, AS NON-CONTAGIOUS, TAKE YOUR PICK, BUT I SAID IT LONG AGO WHEN USDA OIE ET AL MADE ATYPICAL SCRAPIE A LEGAL TRADING COMMODITY, I SAID YOUR PUTTING THE CART BEFORE THE HORSE, AND THAT'S EXACTLY WHAT THEY DID, and it's called in Texas, TEXAS TSE PRION HOLDEM POKER, WHO'S ALL IN $$$
***> AS is considered more likely (subjective probability range 50–66%) that AS is a non-contagious, rather than a contagious, disease.
SNIP...SEE;
THURSDAY, JULY 8, 2021
EFSA Scientific report on the analysis of the 2‐year compulsory intensified monitoring of atypical scrapie
RE-Inactivation of porcine endogenous retrovirus in pigs using CPISPR-Cas9
seems that the USA feed ban for ruminant protein is still a serious problem, so there seems to still be a risk factor for pigs and Transmissible Spongiform Encephalopathy TSE prion disease. now with the updated science showing that pigs are susceptible to the Chronic Wasting Disease TSE Prion ORALLY, and cwd running rampant in the USA, any use of porcine organs should be tested for the CWD TSE Prion...
Research Project: TRANSMISSION, DIFFERENTIATION, AND PATHOBIOLOGY OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHIES
Location: Virus and Prion Research
Title: Disease-associated prion protein detected in lymphoid tissues from pigs challenged with the agent of chronic wasting disease
Author item Moore, Sarah item Kunkle, Robert item Kondru, Naveen item Manne, Sireesha item Smith, Jodi item Kanthasamy, Anumantha item West Greenlee, M item Greenlee, Justin
Submitted to: Prion Publication Type: Abstract Only Publication Acceptance Date: 3/15/2017
Publication Date: N/A Citation: N/A Interpretive Summary:
Technical Abstract: Aims: Chronic wasting disease (CWD) is a naturally-occurring, fatal neurodegenerative disease of cervids. We previously demonstrated that disease-associated prion protein (PrPSc) can be detected in the brain and retina from pigs challenged intracranially or orally with the CWD agent. In that study, neurological signs consistent with prion disease were observed only in one pig: an intracranially challenged pig that was euthanized at 64 months post-challenge. The purpose of this study was to use an antigen-capture immunoassay (EIA) and real-time quaking-induced conversion (QuIC) to determine whether PrPSc is present in lymphoid tissues from pigs challenged with the CWD agent.
Methods: At two months of age, crossbred pigs were challenged by the intracranial route (n=20), oral route (n=19), or were left unchallenged (n=9). At approximately 6 months of age, the time at which commercial pigs reach market weight, half of the pigs in each group were culled (<6 month challenge groups). The remaining pigs (>6 month challenge groups) were allowed to incubate for up to 73 months post challenge (mpc). The retropharyngeal lymph node (RPLN) was screened for the presence of PrPSc by EIA and immunohistochemistry (IHC). The RPLN, palatine tonsil, and mesenteric lymph node (MLN) from 6-7 pigs per challenge group were also tested using EIA and QuIC.
Results: PrPSc was not detected by EIA and IHC in any RPLNs. All tonsils and MLNs were negative by IHC, though the MLN from one pig in the oral <6 month group was positive by EIA. PrPSc was detected by QuIC in at least one of the lymphoid tissues examined in 5/6 pigs in the intracranial <6 months group, 6/7 intracranial >6 months group, 5/6 pigs in the oral <6 months group, and 4/6 oral >6 months group. Overall, the MLN was positive in 14/19 (74%) of samples examined, the RPLN in 8/18 (44%), and the tonsil in 10/25 (40%). Conclusions:
This study demonstrates that PrPSc accumulates in lymphoid tissues from pigs challenged intracranially or orally with the CWD agent, and can be detected as early as 4 months after challenge.
CWD-infected pigs rarely develop clinical disease and if they do, they do so after a long incubation period. This raises the possibility that CWD-infected pigs could shed prions into their environment long before they develop clinical disease.
Furthermore, lymphoid tissues from CWD-infected pigs could present a potential source of CWD infectivity in the animal and human food chains.
CONFIDENTIAL
EXPERIMENTAL PORCINE SPONGIFORM ENCEPHALOPATHY
While this clearly is a cause for concern we should not jump to the conclusion that this means that pigs will necessarily be infected by bone and meat meal fed by the oral route as is the case with cattle. ...
we cannot rule out the possibility that unrecognised subclinical spongiform encephalopathy could be present in British pigs though there is no evidence for this: only with parenteral/implantable pharmaceuticals/devices is the theoretical risk to humans of sufficient concern to consider any action.
Our records show that while some use is made of porcine materials in medicinal products, the only products which would appear to be in a hypothetically ''higher risk'' area are the adrenocorticotrophic hormone for which the source material comes from outside the United Kingdom, namely America China Sweden France and Germany. The products are manufactured by Ferring and Armour. A further product, ''Zenoderm Corium implant'' manufactured by Ethicon, makes use of porcine skin - which is not considered to be a ''high risk'' tissue, but one of its uses is described in the data sheet as ''in dural replacement''. This product is sourced from the United Kingdom.....
snip...see much more here ;
OIE Bulletin
Camel prion disease: a possible emerging disease in dromedary camel populations?
The identification of a new prion disease in dromedary camels in Algeria and Tunisia, called camel prion disease (CPD), extends the spectrum of animal species naturally susceptible to prion diseases and opens up new research areas for investigation.
Camel prion disease was identified in 2018 in adult camels showing clinical signs at the ante mortem inspection at slaughterhouses in the region of Ouargla (Algeria), and in 2019 in the region of Tataouine (Tunisia). It adds to the group of existing animal prion diseases, including scrapie in sheep and goats, chronic wasting disease (CWD) in cervids and BSE (mainly in bovines). The detection of a new prion disease in the dromedary population requires attention and investigation needs to be carried out to assess the risks of this disease to animal and public health. As of today, very limited epidemiological information is available to assess the prevalence, geographical distribution and dynamic of the transmission of the disease.
Based on the clinical signs suggesting prion disease, CPD seems to have occurred in 3.1% of the dromedaries brought to the abattoir in Ouargla. Pathognomonic neurodegeneration and disease specific prion protein (PrPSc) were detected in brain tissue from three symptomatic animals (source:
In May 2019, the OIE received a report from Tunisia on a single case of a 12-year-old slaughtered dromedary camel showing neurological signs confirmed as CPD by the Istituto Superiore di Sanità (ISS) based in Italy.
©B. Babelhadj/University Kasdi Merbah, Algeria
2
Is camel prion disease transmissible in natural conditions?
The involvement of lymphoid tissue in prion replication, observed both in the Algeria and Tunisia cases, is suggestive of a peripheral pathogenesis, which is thought to be a prerequisite for prion shedding into the environment. As with other animal prion diseases, such as scrapie and CWD, in which lymphoid tissues are extensively involved and horizontal transmission occurs efficiently under natural conditions, the detection of prion proteins in lymph nodes is suggestive of the infectious nature of CPD and concurs to hypothesise the potential impact of CPD on animal health. No evidence is currently available with which to argue for the relevance of CPD for human health. However, no absolute species barrier exists in prion diseases and minimising the exposure of humans to prion-infected animal products is an essential aspect of public health protection. As for the relationship between CPD and other animal prion diseases, preliminary analyses suggest that CPD prions have a different molecular signature from scrapie and BSE.
Actions on the follow up of CPD
Since the first description of CPD, the OIE promoted discussions on the impact of this new disease through the OIE Scientific Commission for Animal Diseases (Scientific Commission). The Scientific Commission consulted two OIE ad hoc Groups, one on BSE risk status evaluation of Members and the other on camelids. It analysed the information available from the Algeria and Tunisia cases to evaluate if CPD should be considered an ‘emerging disease’ based on the criteria listed in the Terrestrial Animal Health Code1 .
The OIE Scientific Commission noted that limited surveillance data were available on the prevalence of CPD and that the evidence was not sufficient to measure, at that time, the impact of the disease on animal or public health. Therefore, it was concluded that, with the current knowledge, CPD did not currently meet the criteria to be considered an emerging disease. Nonetheless, it was emphasised that CPD should be considered as a new disease not to be overlooked and called for the collection of further scientific evidence through research and surveillance in the affected countries and in countries with dromedary camel populations to measure the impact of the disease. As new scientific evidence becomes available, the OIE Scientific Commission will reassess whether this disease should be considered as an emerging disease.
The worldwide camel population is ~35 million head (FAO, 2019), 88% of which is found in Africa. The camel farming system is evolving rapidly, and these animals represent vital sources of meat, milk and transportation for millions of people living in the most arid regions of the world. This makes it necessary to assess the risk for animal and human health and to develop evidence-based policies to control and limit the spread of the disease in animals, and to minimise human exposure. As a first step, the awareness of Veterinary Services about CPD and its diagnostic capacity needs to be improved in all countries where dromedaries are part of the domestic livestock.
At the regional level, CPD was first discussed in the 18th Joint Permanent Committee of the Mediterranean Animal Health Network (REMESA) held in Cairo, Egypt, in June 2019 where an expert 1 a new occurrence in an animal of a disease, infection or infestation, causing a significant impact on animal or public health resulting from a) a change of a known pathogenic agent or its spread to a new geographic area or species, or b) a previously unrecognised pathogenic agent or disease diagnosed for the first time www.oiebulletin.com
3
from ISS, Italy, shared the knowledge available on the new disease with the 15 REMESA Member Countries. The discussion highlighted the need to strengthen surveillance systems in order to collect epidemiological data to inform the risk assessments. The results of these risk assessments will support the implementation of evidence-based policies to manage the risks in both animals and humans.
CPD was recently discussed atthe 15thConference of the OIE Regional Commission for the Middle East in November. During this conference, the CAMENET (Camel Middle East Network) launched a wide ranging proposal for training, coordinated surveillance and research on CPD. In addition, the ERFAN (Enhancing Research for Africa Network), a platform aimed at enhancing scientific cooperation between Africa and Italy, during its 2nd ERFAN meeting for North Africa, presented a project on CPD with the objective of increasing CPD coordinated surveillance in North Africa.
The OIE, through its Reference Laboratories for prion diseases, and by involving the above scientific initiatives, is keeping a close watch on the evolution of the disease to gather scientific evidence and to allow a proper and more thorough assessment of the risk associated with this novel disease.
◼ December 2019
Very low oral exposure to prions of brain or saliva origin can transmit chronic wasting disease
Nathaniel D. Denkers ,Clare E. Hoover ,Kristen A. Davenport,Davin M. Henderson,Erin E. McNulty,Amy V. Nalls,Candace K. Mathiason,Edward A. Hoover
Published: August 20, 2020
We report that oral exposure to as little as 300 nanograms (ng) of CWD-positive brain or to saliva containing seeding activity equivalent to 300 ng of CWD-positive brain, were sufficient to transmit CWD disease. This was true whether the inoculum was administered as a single bolus or divided as three weekly 100 ng exposures. However, when the 300 ng total dose was apportioned as 10, 30 ng doses delivered over 12 weeks, no infection occurred. While low-dose exposures to prions of brain or saliva origin prolonged the time from inoculation to first detection of infection, once infection was established, we observed no differences in disease pathogenesis. These studies suggest that the CWD minimum infectious dose approximates 100 to 300 ng CWD-positive brain (or saliva equivalent), and that CWD infection appears to conform more with a threshold than a cumulative dose dynamic.
WE know now, and we knew decades ago, that 5.5 grams of suspect feed in TEXAS was enough to kill 100 cows.
look at the table and you'll see that as little as 1 mg (or 0.001 gm) caused 7% (1 of 14) of the cows to come down with BSE;
Risk of oral infection with bovine spongiform encephalopathy agent in primates
Corinne Ida Lasmézas, Emmanuel Comoy, Stephen Hawkins, Christian Herzog, Franck Mouthon, Timm Konold, Frédéric Auvré, Evelyne Correia, Nathalie Lescoutra-Etchegaray, Nicole Salès, Gerald Wells, Paul Brown, Jean-Philippe Deslys
Summary The uncertain extent of human exposure to bovine spongiform encephalopathy (BSE)--which can lead to variant Creutzfeldt-Jakob disease (vCJD)--is compounded by incomplete knowledge about the efficiency of oral infection and the magnitude of any bovine-to-human biological barrier to transmission. We therefore investigated oral transmission of BSE to non-human primates. We gave two macaques a 5 g oral dose of brain homogenate from a BSE-infected cow. One macaque developed vCJD-like neurological disease 60 months after exposure, whereas the other remained free of disease at 76 months. On the basis of these findings and data from other studies, we made a preliminary estimate of the food exposure risk for man, which provides additional assurance that existing public health measures can prevent transmission of BSE to man.
snip...
BSE bovine brain inoculum
100 g 10 g 5 g 1 g 100 mg 10 mg 1 mg 0·1 mg 0·01 mg
Primate (oral route)* 1/2 (50%)
Cattle (oral route)* 10/10 (100%) 7/9 (78%) 7/10 (70%) 3/15 (20%) 1/15 (7%) 1/15 (7%)
RIII mice (ic ip route)* 17/18 (94%) 15/17 (88%) 1/14 (7%)
PrPres biochemical detection
The comparison is made on the basis of calibration of the bovine inoculum used in our study with primates against a bovine brain inoculum with a similar PrPres concentration that was inoculated into mice and cattle.8 *Data are number of animals positive/number of animals surviving at the time of clinical onset of disease in the first positive animal (%). The accuracy of bioassays is generally judged to be about plus or minus 1 log. ic ip=intracerebral and intraperitoneal.
Table 1: Comparison of transmission rates in primates and cattle infected orally with similar BSE brain inocula
Published online January 27, 2005
It is clear that the designing scientists must
also have shared Mr Bradley’s surprise at the results because all the dose
levels right down to 1 gram triggered infection.
6. It also appears to me that Mr Bradley’s answer (that it would take less than say 100
grams) was probably given with the benefit of hindsight; particularly if one
considers that later in the same answer Mr Bradley expresses his surprise that it
could take as little of 1 gram of brain to cause BSE by the oral route within the
same species. This information did not become available until the "attack rate"
experiment had been completed in 1995/96. This was a titration experiment
designed to ascertain the infective dose. A range of dosages was used to ensure
that the actual result was within both a lower and an upper limit within the study
and the designing scientists would not have expected all the dose levels to trigger
infection. The dose ranges chosen by the most informed scientists at that time
ranged from 1 gram to three times one hundred grams. It is clear that the designing
scientists must have also shared Mr Bradley’s surprise at the results because all the
dose levels right down to 1 gram triggered infection.
***> cattle, pigs, sheep, cwd, tse, prion, oh my!
***> In contrast, cattle are highly susceptible to white-tailed deer CWD and mule deer CWD in experimental conditions but no natural CWD infections in cattle have been reported (Sigurdson, 2008; Hamir et al., 2006).
Sheep and cattle may be exposed to CWD via common grazing areas with affected deer but so far, appear to be poorly susceptible to mule deer CWD (Sigurdson, 2008). In contrast, cattle are highly susceptible to white-tailed deer CWD and mule deer CWD in experimental conditions but no natural CWD infections in cattle have been reported (Sigurdson, 2008; Hamir et al., 2006). It is not known how susceptible humans are to CWD but given that the prion can be present in muscle, it is likely that humans have been exposed to the agent via consumption of venison (Sigurdson, 2008). Initial experimental research suggests that human susceptibility to CWD is low and there may be a robust species barrier for CWD transmission to humans (Sigurdson, 2008), however the risk appetite for a public health threat may still find this level unacceptable.
DEFRA
Friday, December 14, 2012
DEFRA U.K. What is the risk of Chronic Wasting Disease CWD being introduced into Great Britain? A Qualitative Risk Assessment October 2012
snip.....
In the USA, under the Food and Drug Administration's BSE Feed Regulation (21 CFR 589.2000) most material (exceptions include milk, tallow, and gelatin) from deer and elk is prohibited for use in feed for ruminant animals. With regards to feed for non-ruminant animals, under FDA law, CWD positive deer may not be used for any animal feed or feed ingredients. For elk and deer considered at high risk for CWD, the FDA recommends that these animals do not enter the animal feed system. However, this recommendation is guidance and not a requirement by law. Animals considered at high risk for CWD include:
1) animals from areas declared to be endemic for CWD and/or to be CWD eradication zones and
2) deer and elk that at some time during the 60-month period prior to slaughter were in a captive herd that contained a CWD-positive animal.
Therefore, in the USA, materials from cervids other than CWD positive animals may be used in animal feed and feed ingredients for non-ruminants.
The amount of animal PAP that is of deer and/or elk origin imported from the USA to GB can not be determined, however, as it is not specified in TRACES.
It may constitute a small percentage of the 8412 kilos of non-fish origin processed animal proteins that were imported from US into GB in 2011.
Overall, therefore, it is considered there is a __greater than negligible risk___ that (nonruminant) animal feed and pet food containing deer and/or elk protein is imported into GB.
There is uncertainty associated with this estimate given the lack of data on the amount of deer and/or elk protein possibly being imported in these products.
snip.....
36% in 2007 (Almberg et al., 2011). In such areas, population declines of deer of up to 30 to 50% have been observed (Almberg et al., 2011). In areas of Colorado, the prevalence can be as high as 30% (EFSA, 2011). The clinical signs of CWD in affected adults are weight loss and behavioural changes that can span weeks or months (Williams, 2005). In addition, signs might include excessive salivation, behavioural alterations including a fixed stare and changes in interaction with other animals in the herd, and an altered stance (Williams, 2005). These signs are indistinguishable from cervids experimentally infected with bovine spongiform encephalopathy (BSE). Given this, if CWD was to be introduced into countries with BSE such as GB, for example, infected deer populations would need to be tested to differentiate if they were infected with CWD or BSE to minimise the risk of BSE entering the human food-chain via affected venison. snip..... The rate of transmission of CWD has been reported to be as high as 30% and can approach 100% among captive animals in endemic areas (Safar et al., 2008).
snip.....
In summary, in endemic areas, there is a medium probability that the soil and surrounding environment is contaminated with CWD prions and in a bioavailable form. In rural areas where CWD has not been reported and deer are present, there is a greater than negligible risk the soil is contaminated with CWD prion. snip..... In summary, given the volume of tourists, hunters and servicemen moving between GB and North America, the probability of at least one person travelling to/from a CWD affected area and, in doing so, contaminating their clothing, footwear and/or equipment prior to arriving in GB is greater than negligible... For deer hunters, specifically, the risk is likely to be greater given the increased contact with deer and their environment. However, there is significant uncertainty associated with these estimates.
snip.....
Therefore, it is considered that farmed and park deer may have a higher probability of exposure to CWD transferred to the environment than wild deer given the restricted habitat range and higher frequency of contact with tourists and returning GB residents.
snip.....
a review of banned mad cow feed in USA;
BANNED MAD COW FEED IN COMMERCE IN ALABAMA
Date: September 6, 2006 at 7:58 am PST PRODUCT
a) EVSRC Custom dairy feed, Recall # V-130-6;
b) Performance Chick Starter, Recall # V-131-6;
c) Performance Quail Grower, Recall # V-132-6;
d) Performance Pheasant Finisher, Recall # V-133-6.
CODE None RECALLING FIRM/MANUFACTURER Donaldson & Hasenbein/dba J&R Feed Service, Inc., Cullman, AL, by telephone on June 23, 2006 and by letter dated July 19, 2006. Firm initiated recall is complete.
REASON
Dairy and poultry feeds were possibly contaminated with ruminant based protein.
VOLUME OF PRODUCT IN COMMERCE 477.72 tons
DISTRIBUTION AL
______________________________
http://www.fda.gov/bbs/topics/enforce/2006/ENF00968.html
PRODUCT Bulk custom dairy pre-mixes,
Recall # V-120-6 CODE None RECALLING FIRM/MANUFACTURER Ware Milling Inc., Houston, MS, by telephone on June 23, 2006. Firm initiated recall is complete. REASON Possible contamination of dairy animal feeds with ruminant derived meat and bone meal.
VOLUME OF PRODUCT IN COMMERCE 350 tons
DISTRIBUTION AL and MS
______________________________
PRODUCT
a) Tucker Milling, LLC Tm 32% Sinking Fish Grower, #2680-Pellet, 50 lb. bags, Recall # V-121-6;
b) Tucker Milling, LLC #31120, Game Bird Breeder Pellet, 50 lb. bags, Recall # V-122-6;
c) Tucker Milling, LLC #31232 Game Bird Grower, 50 lb. bags, Recall # V-123-6;
d) Tucker Milling, LLC 31227-Crumble, Game Bird Starter, BMD Medicated, 50 lb bags, Recall # V-124-6;
e) Tucker Milling, LLC #31120, Game Bird Breeder, 50 lb bags, Recall # V-125-6;
f) Tucker Milling, LLC #30230, 30 % Turkey Starter, 50 lb bags, Recall # V-126-6;
g) Tucker Milling, LLC #30116, TM Broiler Finisher, 50 lb bags, Recall # V-127-6
CODE All products manufactured from 02/01/2005 until 06/20/2006 RECALLING FIRM/MANUFACTURER Recalling Firm: Tucker Milling LLC, Guntersville, AL, by telephone and visit on June 20, 2006, and by letter on June 23, 2006. Manufacturer: H. J. Baker and Brothers Inc., Stamford, CT. Firm initiated recall is ongoing.
REASON Poultry and fish feeds which were possibly contaminated with ruminant based protein were not labeled as "Do not feed to ruminants".
VOLUME OF PRODUCT IN COMMERCE 7,541-50 lb bags
DISTRIBUTION AL, GA, MS, and TN
END OF ENFORCEMENT REPORT FOR AUGUST 9, 2006
###
http://www.fda.gov/bbs/topics/ENFORCE/2006/ENF00964.html
Subject: MAD COW FEED RECALL AL AND FL VOLUME OF PRODUCT IN COMMERCE 125 TONS Products manufactured from 02/01/2005 until 06/06/2006
Date: August 6, 2006 at 6:16 pm PST PRODUCT
a) CO-OP 32% Sinking Catfish, Recall # V-100-6;
b) Performance Sheep Pell W/Decox/A/N, medicated, net wt. 50 lbs, Recall # V-101-6;
c) Pro 40% Swine Conc Meal -- 50 lb, Recall # V-102-6;
d) CO-OP 32% Sinking Catfish Food Medicated, Recall # V-103-6;
e) "Big Jim's" BBB Deer Ration, Big Buck Blend, Recall # V-104-6;
f) CO-OP 40% Hog Supplement Medicated Pelleted, Tylosin 100 grams/ton, 50 lb. bag, Recall # V-105-6;
g) Pig Starter Pell II, 18% W/MCDX Medicated 282020, Carbadox -- 0.0055%, Recall # V-106-6;
h) CO-OP STARTER-GROWER CRUMBLES, Complete Feed for Chickens from Hatch to 20 Weeks, Medicated, Bacitracin Methylene Disalicylate, 25 and 50 Lbs, Recall # V-107-6;
i) CO-OP LAYING PELLETS, Complete Feed for Laying Chickens, Recall # 108-6;
j) CO-OP LAYING CRUMBLES, Recall # V-109-6;
k) CO-OP QUAIL FLIGHT CONDITIONER MEDICATED, net wt 50 Lbs, Recall # V-110-6;
l) CO-OP QUAIL STARTER MEDICATED, Net Wt. 50 Lbs, Recall # V-111-6;
m) CO-OP QUAIL GROWER MEDICATED, 50 Lbs, Recall # V-112-6 CODE
Product manufactured from 02/01/2005 until 06/06/2006
RECALLING FIRM/MANUFACTURER Alabama Farmers Cooperative, Inc., Decatur, AL, by telephone, fax, email and visit on June 9, 2006. FDA initiated recall is complete.
REASON Animal and fish feeds which were possibly contaminated with ruminant based protein not labeled as "Do not feed to ruminants".
VOLUME OF PRODUCT IN COMMERCE 125 tons
DISTRIBUTION AL and FL
END OF ENFORCEMENT REPORT FOR AUGUST 2, 2006
###
http://www.fda.gov/bbs/topics/enforce/2006/ENF00963.html
MAD COW FEED RECALL USA EQUALS 10,878.06 TONS NATIONWIDE Sun Jul 16, 2006 09:22 71.248.128.67
RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINE -- CLASS II
______________________________
PRODUCT
a) PRO-LAK, bulk weight, Protein Concentrate for Lactating Dairy Animals, Recall # V-079-6;
b) ProAmino II, FOR PREFRESH AND LACTATING COWS, net weight 50lb (22.6 kg), Recall # V-080-6;
c) PRO-PAK, MARINE & ANIMAL PROTEIN CONCENTRATE FOR USE IN ANIMAL FEED, Recall # V-081-6;
d) Feather Meal, Recall # V-082-6 CODE
a) Bulk
b) None
c) Bulk
d) Bulk
RECALLING FIRM/MANUFACTURER H. J. Baker & Bro., Inc., Albertville, AL, by telephone on June 15, 2006 and by press release on June 16, 2006. Firm initiated recall is ongoing.
REASON
Possible contamination of animal feeds with ruminent derived meat and bone meal.
VOLUME OF PRODUCT IN COMMERCE 10,878.06 tons
DISTRIBUTION Nationwide
END OF ENFORCEMENT REPORT FOR July 12, 2006
###
http://www.fda.gov/bbs/topics/enforce/2006/ENF00960.html
10,000,000+ LBS. of PROHIBITED BANNED MAD COW FEED I.E. BLOOD LACED MBM IN COMMERCE USA 2007
Date: March 21, 2007 at 2:27 pm PST
RECALLS AND FIELD CORRECTIONS: VETERINARY MEDICINES -- CLASS II
___________________________________
PRODUCT
Bulk cattle feed made with recalled Darling's 85% Blood Meal, Flash Dried, Recall # V-024-2007
CODE
Cattle feed delivered between 01/12/2007 and 01/26/2007
RECALLING FIRM/MANUFACTURER
Pfeiffer, Arno, Inc, Greenbush, WI. by conversation on February 5, 2007.
Firm initiated recall is ongoing.
REASON
Blood meal used to make cattle feed was recalled because it was cross- contaminated with prohibited bovine meat and bone meal that had been manufactured on common equipment and labeling did not bear cautionary BSE statement.
VOLUME OF PRODUCT IN COMMERCE
42,090 lbs.
DISTRIBUTION
WI
___________________________________
PRODUCT
Custom dairy premix products: MNM ALL PURPOSE Pellet, HILLSIDE/CDL Prot- Buffer Meal, LEE, M.-CLOSE UP PX Pellet, HIGH DESERT/ GHC LACT Meal, TATARKA, M CUST PROT Meal, SUNRIDGE/CDL PROTEIN Blend, LOURENZO, K PVM DAIRY Meal, DOUBLE B DAIRY/GHC LAC Mineral, WEST PIONT/GHC CLOSEUP Mineral, WEST POINT/GHC LACT Meal, JENKS, J/COMPASS PROTEIN Meal, COPPINI - 8# SPECIAL DAIRY Mix, GULICK, L-LACT Meal (Bulk), TRIPLE J - PROTEIN/LACTATION, ROCK CREEK/GHC MILK Mineral, BETTENCOURT/GHC S.SIDE MK-MN, BETTENCOURT #1/GHC MILK MINR, V&C DAIRY/GHC LACT Meal, VEENSTRA, F/GHC LACT Meal, SMUTNY, A- BYPASS ML W/SMARTA, Recall # V-025-2007
CODE
The firm does not utilize a code - only shipping documentation with commodity and weights identified.
RECALLING FIRM/MANUFACTURER
Rangen, Inc, Buhl, ID, by letters on February 13 and 14, 2007. Firm initiated recall is complete.
REASON
Products manufactured from bulk feed containing blood meal that was cross contaminated with prohibited meat and bone meal and the labeling did not bear cautionary BSE statement.
VOLUME OF PRODUCT IN COMMERCE
9,997,976 lbs.
DISTRIBUTION
ID and NV
END OF ENFORCEMENT REPORT FOR MARCH 21, 2007
http://www.fda.gov/Safety/Recalls/EnforcementReports/2007/ucm120446.htm
MONDAY, OCTOBER 10, 2022
Docket No: 2002N-0273 (formerly Docket No. 02N-0273) Substances Prohibited From Use in Animal Food and Feed Scientists Comments December 20, 2005
This information is critical, and should continue to be collected.
The TSE prion is spreading across the USA in Cervid as in CWD TSE Prion.
The mad cow surveillance, feed ban, testing, and SRM removal there from, has been, and still is, a terrible failure.
WE know that the USA Food and Drug Administration's BSE Feed Regulation (21 CFR 589.2000) of August 1997 was/is a colossal failure, and proven to be so year after year, decade after decade, and this was just admitted by the FDA et al (see below FDA Reports on VFD Compliance Sept. 2019 report).
God, all these decades you hear from all the warning letters on SRM that were released to the public for consumption, that even if they did eat a SRM, the BSE Feed Regulation (21 CFR 589.2000) of August 1997 would save that tissue from that animal from having a TSE Prion, was nothing but lies. what about those children all across the USA that were fed the most high risk cattle for mad cow disease, i.e. dead stock downer cows via the USDA School lunch program, who will watch those kids for the next 50 years for cjd tse prion aka mad cow disease, let alone all the folks consuming SRMs that have been exposed to mad cow type disease in different livestock species, due to the fact the USA colossal failure of the BSE Feed Regulation (21 CFR 589.2000) of August 1997. it's all documented below, see for yourself; SUNDAY, SEPTEMBER 1, 2019 FDA Reports on VFD Compliance
Tuesday, September 10, 2019
FSIS [Docket No. FSIS–2019–0021] Notice of Request To Renew an Approved Information Collection: Specified Risk Materials Singeltary Submission
-------- Original Message --------
Subject: re-BSE prions propagate as either variant CJD-like or sporadic CJD
Date: Thu, 28 Nov 2002 10:23:43 -0000
From: "Asante, Emmanuel A" e.asante@ic.ac.uk
To: "'flounder@wt.net'" flounder@wt.net
Dear Terry,
I have been asked by Professor Collinge to respond to your request. I am a Senior Scientist in the MRC Prion Unit and the lead author on the paper. I have attached a pdf copy of the paper for your attention.
Thank you for your interest in the paper.
In respect of your first question, the simple answer is, ***yes. As you will find in the paper, we have managed to associate the alternate phenotype to type 2 PrPSc, the commonest sporadic CJD. It is too early to be able to claim any further sub-classification in respect of Heidenhain variant CJD or Vicky Rimmer's version. It will take further studies, which are on-going, to establish if there are sub-types to our initial finding which we are now reporting. The main point of the paper is that, as well as leading to the expected new variant CJD phenotype, BSE transmission to the 129-methionine genotype can lead to an alternate phenotype which is indistinguishable from type 2 PrPSc.
I hope reading the paper will enlighten you more on the subject. If I can be of any further assistance please to not hesitate to ask. Best wishes.
Emmanuel Asante
<<Asante et al 2002.pdf>>
____________________________________
Dr. Emmanuel A Asante MRC Prion Unit & Neurogenetics Dept. Imperial College School of Medicine (St. Mary's) Norfolk Place, LONDON W2 1PG Tel: +44 (0)20 7594 3794 Fax: +44 (0)20 7706 3272 email: e.asante@ic.ac.uk (until 9/12/02) New e-mail: e.asante@prion.ucl.ac.uk (active from now)
____________________________________
''This study demonstrates that the H-type BSE agent is transmissible by the oronasal route. Cattle with the EK211 genotype are oronasally susceptible to small doses of the H-BSE agent from either EK211 or EE211 (wild type) donors. Wild-type EE211 cattle remained asymptomatic for the duration of the experiment with this small dose (0.1g) of inoculum. These results reinforce the need for ongoing surveillance for classical and atypical BSE to minimize the risk of potentially infectious tissues entering the animal or human food chains.''
***Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility.***
Even if the prevailing view is that sporadic CJD is due to the spontaneous formation of CJD prions, it remains possible that its apparent sporadic nature may, at least in part, result from our limited capacity to identify an environmental origin.
PRION CONFERENCE 2022 ABSTRACTS CWD TSE PrP ZOONOSIS
Transmission of prion infectivity from CWD-infected macaque tissues to rodent models demonstrates the zoonotic potential of chronic wasting disease.
Samia Hannaouia, Ginny Chenga, Wiebke Wemheuerb, Walter J. Schulz-Schaefferb, Sabine Gilcha, and Hermann M. Schätzla aDepartment of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine & Hotchkiss Brain Institute; University of Calgary, Calgary, Canada; bInstitute of Neuropathology, Medical Faculty, Saarland University, Homburg/Saar, Germany
Aims: Chronic wasting disease (CWD) is a prion disease of cervids. Its rapid geographic expansion, shedding of infectivity and persistence in the environment for many years are of concern for humans. Here, we provide the first evidence by transmission experiments to different transgenic mouse models and bank voles that Cynomolgus macaques inoculated via different routes with CWD-positive cervid tissues harbor infectious prions that elicit clinical disease in rodents.
Material and Methods: We used tissue materials from macaques inoculated with CWD to inoculate transgenic mice overexpressing cervid PrPCfollowed by transmission into bank voles. We used RT-QuIC, immunoblot and PET blot analysis to assess brains, spinal cords, and tissues of the gastrointestinal tract (GIT) for the presence of prions.
Results: Our results show that of the macaque materials that induced clinical disease in transgenic mice,73% were from the CNS (46% spinal cord and 27% brain), and 27% were from the spleen, although attack rates were low around 20%. Clinical mice did not display PK-resistant PrPSc(PrPres) in immunoblot, but showed low-levels of prion seeding activity. Transmission into bank voles from clinical transgenic mice led to a 100% attack rate with typical PrPressignature in immunoblot, which was different from that of voles inoculated directly with CWD or scrapie prions. High-level prion seeding activity in brain and spinal cord and PrPresdeposition in the brain were present. Remarkably, we also found prion seeding activity in GIT tissues of inoculated voles. Second passage in bank voles led to a 100% attack rate in voles inoculated with brain, spinal cord and small intestine material from first round animals, with PrPresin immunoblot, prion seeding activity, and PrPresdeposition in the brain. Shortened survival times indicate adaptation in the new host. This also shows that prions detected in GIT tissues are infectious and transmissible. Transmission of brain material from sick voles back to cervidized mice revealed transmission in these mice with a 100% attack rate, and interestingly, with different biochemical signature and distribution in the brain.
Conclusions: Our findings demonstrate that macaques, considered the best model for the zoonotic potential of prions, were infected upon CWD challenge, including oral one. The disease manifested as atypical in macaques and transgenic mice, but with infectivity present at all times, as unveiled in the bank vole model with an unusual tissue tropism.
Funded by: The National Institutes of Health, USA, and the Alberta Prion Research Institute/Alberta Innovates Canada. Grant number: 1R01NS121016-01; 201,600,023
Acknowledgement: We thank Umberto Agrimi, Istituto Superiore di Sanità, Rome, Italy, and Michael Beekes, Robert-Koch Institute Berlin, Germany, for providing the bank vole model. We thank the University of Calgary animal facility staff and Dr. Stephanie Anderson for animal care.
Transmission of Cervid Prions to Humanized Mice Demonstrates the Zoonotic Potential of CWD
Samia Hannaouia, Irina Zemlyankinaa, Sheng Chun Changa, Maria Immaculata Arifina, Vincent Béringueb, Debbie McKenziec, Hermann M. Schatzla, and Sabine Gilcha
aDepartment of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine; Hotchkiss Brain Institute; University of Calgary, Calgary, Canada; bUniversité Paris-Saclay, INRAE, UVSQ, VIM, Jouy-en-Josas, France; cDepartment of Biological Sciences, Center for Prions and Protein Folding Diseases, University of Alberta, Edmonton, Canada
Aims: Chronic wasting disease (CWD), a prion disease of cervids, spreads efficiently among wild and farmed animals. Potential transmission to humans of CWD is a growing concern due to its increasing prevalence. Here, we aimed to determine the zoonotic potential of CWD using a mouse model for human prion diseases.
Material and Methods: Transgenic mice overexpressing human PrPChomozygous for methionine at codon 129 (tg650) were inoculated intracerebrally with brain homogenates of white-tailed deer infected with Wisc-1/CWD1 or 116AG CWD strains. Mice were monitored for clinical signs and were euthanized at terminal disease. Brains were tested by RT-QuIC, western blot upon PK digestion, and immunohistochemistry; fecal homogenates were analyzed by RT-QuIC. Brain/spinal cord and fecal homogenates of CWD-inoculated tg650 mice were inoculated into tg650 mice or bank voles. Brain homogenates of bank voles inoculated with fecal homogenates of CWD-infected tg650 mice were used for second passage in bank voles.
Results: Here, we provide the strongest evidence supporting the zoonotic potential of CWD prions, and their possible phenotype in humans. Inoculation of mice expressing human PrPCwith deer CWD isolates (strains Wisc-1 and 116AG) resulted in atypical clinical manifestations in > 75% of the mice, with myoclonus as leading clinical sign. Most of tg650 brain homogenates were positive for seeding activity in RT-QuIC. Clinical disease and presentation was transmissible to tg650 mice and bank voles. Intriguingly, protease-resistant PrP in the brain of tg650 mice resembled that found in a familial human prion disease and was transmissible upon passage. Abnormal PrP aggregates upon infection with Wisc-1 were detectable in thalamus, hypothalamus, and midbrain/pons regions.
Unprecedented in human prion disease, feces of CWD-inoculated tg650 mice harbored prion seeding activity and infectious prions, as shown by inoculation of bank voles and tg650 with fecal homogenates.
Conclusions: This is the first evidence that CWD can infect humans and cause disease with a distinctive clinical presentation, signature, and tropism, which might be transmissible between humans while current diagnostic assays might fail to detect it. These findings have major implications for public health and CWD-management.
Funded by: We are grateful for financial support from the Natural Sciences and Engineering Research Council of Canada, the National Institutes of Health, Genome Canada, and the Alberta Prion Research Institute. SG is supported by the Canada Research Chairs program.
Acknowledgement: We thank Dr. Trent Bollinger, WCVM, University of Saskatchewan, Saskatoon, Canada, for providing brain tissue from the WTD-116AG isolate, Dr. Stéphane Haïk, ICM, Paris, France, for providing brain tissue from vCJD and sCJD cases, and Dr. Umberto Agrimi, Istituto Superiore di Sanità, Italy, for the bank vole model. We thank animal facility staff for animal care, Dr. Stephanie Anderson for veterinary oversight, and Yo-Ching Cheng for preparing recombinant PrP substrates. Thank you to Dr. Stephanie Booth and Jennifer Myskiw, Public Health Agency of Canada, Canada.
The chronic wasting disease agent from white-tailed deer is infectious to humanized mice after passage through raccoons
Eric Cassmanna, Xu Qib, Qingzhong Kongb, and Justin Greenleea
aNational Animal Disease Center, Agricultural Research Service, US Department of Agriculture, Ames, IA, USA bDepartments of Pathology, Neurology, National Center for Regenerative Medicine, and National Prion Disease Pathology Surveillance Center, Case Western Reserve University, Cleveland, Ohio, USA
Aims: Evaluate the zoonotic potential of the raccoon passaged chronic wasting disease (CWD) agent in humanized transgenic mice in comparison with the North American CWD agent from the original white-tailed deer host.
Material and Methods: Pooled brain material (GG96) from a CWD positive herd was used to oronasally inoculate two white-tailed deer with wild-type prion protein genotype and intracranially inoculate a raccoon. Brain homogenates (10% w/v) from the raccoon and the two white-tailed deer were used to intracranially inoculate separate groups of transgenic mice that express human prion protein with methionine (M) at codon 129 (Tg40h). Brains and spleens were collected from mice at experimental endpoints of clinical disease or approximately 700 days post-inoculation. Tissues were divided and homogenized or fixed in 10% buffered neutral formalin. Immunohistochemistry, enzyme immunoassay, and western blot were used to detect misfolded prion protein (PrPSc) in tissue.
Results: Humanized transgenic mice inoculated with the raccoon passaged CWD agent from white-tailed deer exhibited a 100% (12/12) attack rate with an average incubation period of 605 days. PrPScwas detected in brain tissue by enzyme immunoassay with an average optical density of 3.6/4.0 for positive brains. PrPScalso was detected in brain tissue by western blot and immunohistochemistry. No PrPScwas detected in the spleens of mice inoculated with the raccoon passaged CWD agent. Humanized mice inoculated with the CWD agent from white-tailed deer did not have detectable PrPScusing conventional immunoassay techniques.
Conclusions: The host range of the CWD agent from white-tailed deer was expanded in our experimental model after one passage through raccoons.
Funded by: This research was funded in its entirety by congressionally appropriated funds to the United States Department of Agriculture, Agricultural Research Service. The funders of the work did not influence study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Acknowledgement: We thank Quazetta Brown, Lexi Frese, Rylie Frese, Kevin Hassall, Leisa Mandell, and Trudy Tatum for providing excellent technical support to this project.
Stable and highly zoonotic cervid prion strain is possible
Manuel Camacho, Xu Qi, Liuting Qing, Sydney Smith, Jieji Hu, Wanyun Tao, Ignazio Cali, and Qingzhong Kong Department of Pathology, Case Western Reserve University, Cleveland, USA
Aims: Whether CWD prions can infect humans remains unclear despite the very substantial scale and long history of human exposure of CWD in some areas. Multiple in vitro conversion experiments and in vivo animal studies suggest that the CWD-to-human transmission barrier is not unbreakable. A major public health concern on CWD zoonosis is the emergence of highly zoonotic CWD strains. We aim to address the question of whether highly zoonotic CWD strains are possible.
Material and Methods: We inoculated a few sCJD brain samples into cervidized transgenic mice, which were intended as negative controls for bioassays of brain tissues from sCJD cases who had hunted or consumed vension from CWD-endemic states. Some of these mice became infected and their brain tissues were further examined by serial passages in humanized or cervidized mice.
Results: Passage of sCJDMM1 in transgenic mice expressing elk PrP (Tg12) resulted in a ‘cervidized’ CJD strain that we termed CJDElkPrP. We observed 100% transmission of CJDElkPrPin transgenic mice expressing human PrP (Tg40h). We passaged CJDElkPrPtwo more times in the Tg12 mice. We found that such second and third passage CJDElkPrPprions also led to 100% infection in the Tg40h mice. In contrast, we and others found zero or poor transmission of natural elk CWD isolates in humanized mice, despite that natural elk CWD isolates and CJDElkPrPshare the same elk PrP sequence.
Conclusions: Our data demonstrate that highly zoonotic cervid prion strains are not only possible but also can be stably maintained in cervids and that CWD zoonosis is prion strain-dependent.
Funded by: NIH
Grant number: R01NS052319, R01NS088604, R01NS109532
Acknowledgement: We want to thank the National Prion Disease Pathology Surveillance Center and Drs. Allen Jenny and Katherine O’Rourke for providing the sCJD samples and the CWD samples, respectively.
Adaptation of chronic wasting disease (CWD) prion strains in hosts with different PRNP genotypes
Camilo Duque Velasqueza,c, Elizabeth Triscotta,c, Chiye Kima,c, Diana Morenoa,c, Judd Aikenb,c, and Debbie McKenziea,c
aDepartment of Biological Science, University of Alberta, Edmonton, AB T6G 2G8, Canada; bDepartment of Agriculture, Food & Nutritional Science, University of Alberta, Edmonton, AB T6G 2G8, Canada; cCentre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, AB T6G 2M8, Canada
Aims: The contagious nature of CWD epizootics and the PrPCamino acid variation of cervids (and susceptible sympatric species) guarantee the expansion of prion conformational diversity and selective landscapes where new strains can arise. CWD strains can have novel transmission properties including altered host range that may increase zoonotic risk as circulating strains diversify and evolve. We are characterizing the host adaptability of characterized CWD strains as well as CWD isolates from different cervid species in various enzootic regions.
Material and Methods: Characterized CWD strains as well as a number of isolates from hunter-harvested deer were bioassayed in our rodent panel (transgenic mice expressing cervid alleles G96, S96 and H95-PrPC, elk PrPC, bovine PrPC, and both hamsters and non-transgenic laboratory mice). Strain characteristics were compared using computer based scoring of brain pathology (e.g. PrPCWDbrain distribution), western blot and protein misfolding cyclic amplification (PMCA).
Results: Transmission of various isolates resulted in the selection of strain mixtures in hosts expressing similar PrPC, particularly for polymorphic white-tailed deer and for Norwegian reindeer. As of the second passage, transmission of P153 moose prions from Norway has not resulted in emergence of strains with properties similar to any North American CWD strains in our taxonomic collection (Wisc-1, CWD2, H95+and 116AG).
Conclusions: Our data indicates polymorphic white-tailed deer can favor infection with more than one strain. Similar to transmission studies of Colorado CWD isolates from cervids expressing a single PrPCprimary structure, the isolate from Norway reindeer (V214) represents a strain mixture, suggesting intrinsic strain diversity in the Nordfjella epizootic. The diversity of CWD strains with distinct transmission characteristics represents a threat to wildlife, sympatric domestic animals and public health.
Funded by: Genome Canada and Genome Alberta (Alberta Prion Research Institute and Alberta Agriculture & Forestry); NSERC Grant number: #LSARP 10205; NSERC RGPIN-2017-05539
Acknowledgement: We would like to thank Margo Pybus (Alberta Environment and Parks) Trent Bollinger (University of Saskatchewan) for providing us with tissue samples from hunter-harvested deer and Sylvie Benestad for providing moose and reindeer samples.
Application of PMCA to understand CWD prion strains, species barrier and zoonotic potential
Sandra Pritzkowa, Damian Gorskia, Frank Ramireza, Fei Wanga, Glenn C. Tellingb, Justin J. Greenleec, Sylvie L. Benestadd, and Claudio Sotoa aDepartment of Neurology, University of Texas Medical School at Houston, Houston, Texas, USA; bDepartment of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA; cVirus and Prion Research Unit, United States Department of Agriculture, Ames, Iowa, USA; dNorwegian Veterinary Institute, OIE Reference Laboratory for CWD, Ås, Norway
Aims: Chronic wasting disease (CWD) is a prion disease affecting various species of cervids that continues to spread uncontrollably across North America and has recently been detected in Scandinavia (Norway, Sweden and Finland). The mechanisms responsible for the natural transmission of CWD are largely unknown. Furthermore, the risk of CWD transmission to other species, including humans, is also unknown and remains a dangerous enigma. In this study, we investigated the potential of CWD prions to infect several other animal species (sheep, cattle, pig, hamster, and mouse) including humans, by examining their capacity to convert the normal prion protein of distinct species in a PMCA reaction. Moreover, we also investigated whether the in vivo passage of CWD through intermediate species alters their capacity for zoonotic transmission, which may represent a major hazard to human health.
Material and Methods: For these studies, we used brain material from CWD-infected white-tailed deer (Odocoileus virginianus), elk (Cervus canadensis), and mule deer (Odocoileus hemionus) as species native to North America. We also used CWD-infected Moose (Alces alces), reindeer (Rangifer tarandus) and red deer (Cervus elaphus) as Norwegian cervids. We also used brains from cattle, sheep and pigs experimentally infected by CWD. To study interspecies-transmission and zoonotic potential, samples were tested via PMCA for the conversion of PrPCinto PrPScusing different combinations of inoculum and host species. Based on these analyses we estimated the spillover and zoonotic potential for different CWD isolates. We define and quantify spillover and zoonotic potential indices as the efficiency by which CWD prions sustain prion generation in vitro at the expense of normal prion proteins from various mammals and human, respectively.
Results: Our results show that prions from some cervid species, especially those found in Northern Europe, have a higher potential to transmit disease characteristics to other animals. Conversely, CWD-infected cervids originated in North America appear to have a greater potential to generate human PrPSc. We also found that in vivo transmission of CWD to cattle, but not to sheep or pigs substantially increases the ability of these prions to convert human PrPCby PMCA.
Conclusions: Our findings support the existence of different CWD prion strains with distinct spillover and zoonotic potentials. We also conclude that transmission of CWD to other animal species may increase the risk for CWD transmission to humans. Our studies may provide a tool to predict the array of animal species that a given CWD prion could affect and may contribute to understanding the risk of CWD for human health.
Funded by: National Institute of Health Grant number: P01 AI077774
Generation of human chronic wasting disease in transgenic mice
Zerui Wanga, Kefeng Qinb, Manuel V. Camachoa, Ignazio Cali a,c, Jue Yuana, Pingping Shena, Tricia Gillilanda, Syed Zahid Ali Shaha, Maria Gerasimenkoa, Michelle Tanga, Sarada Rajamanickama, Anika Yadatia, Lawrence B. Schonbergerd, Justin Greenleee, Qingzhong Konga,c, James A. Mastriannib, and Wen-Quan Zoua,c
aDepartment of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH, USA; bDepartment of Neurology and Center for Comprehensive Care and Research on Memory Disorders, the University of Chicago Pritzker School of Medicine, Chicago, USA; cNational Prion Disease Pathology Surveillance Center, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA; dDivision of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, 1600 Clifton Rd, Atlanta, GA, USA; eVirus and Prion Research Unit, National Animal Disease Center, USDA, Agricultural Research Service, 1920 Dayton Avenue, Ames, IA, USA
Aims: Chronic wasting disease (CWD) results from the accumulation of an infectious misfolded conformer (PrPSc) of cellular prion protein (PrPC) in the brains of deer and elk. It has been spreading rapidly throughout many regions of North America, exported inadvertently to South Korea, and more recently identified in Europe. Mad cow disease has caused variant Creutzfeldt-Jakob disease (vCJD) in humans and is currently the only known zoonotic prion disease. Whether CWD is transmissible to humans remains uncertain. The aims of our study were not only to confirm whether CWD prion isolates can convert human brain PrPCinto PrPScin vitro by serial protein misfolding cyclic amplification (sPMCA) but also to determine whether the sPMCA-induced CWD-derived human PrPScis infectious.
Material and Methods: Eight CWD prion isolates from 7 elks and 1 deer were used as the seeds while normal human brain homogenates containing either PrP-129 MM (n = 2) or PrP-129 VV (n = 1) were used as the substrates for sPMCA assay. A normal elk brain tissue sample was used as a negative control seed. Two lines of humanized transgenic (Tg) mice expressing either human PrP-129VV or −129 MM polymorphism were included for transmission studies to determine the infectivity of PMCA-amplified PrPSc. Wester blotting and immunohistochemistry and hematoxylin & eosin staining were used for determining PrPScand neuropathological changes of inoculated animals.
Results: We report here the generation of the first CWD-derived infectious human PrPScusing elk CWD PrPScto initiate conversion of human PrPCfrom normal human brain homogenates with PMCA in vitro. Western blotting with a human PrP selective antibody confirmed that the PMCA-generated protease-resistant PrPScwas derived from the human brain PrPCsubstrate. Two lines of humanized transgenic mice expressing human PrPCwith either Val or Met at the polymorphic codon 129 developed clinical prion disease following intracerebral inoculation with the PMCA-generated CWD-derived human PrPSc. Diseased mice exhibited distinct PrPScpatterns and neuropathological changes in the brain.
Conclusions: Our study, using PMCA and animal bioassays, provides the first evidence that CWD PrPSchas the potential to overcome the species barrier and directly convert human PrPCinto infectious PrPScthat can produce bona fide prion disease when inoculated into humanized transgenic mice.
Funded by: CJD Foundation and NIH
Mortality surveillance of persons potentially exposed to chronic wasting disease
R.A. Maddoxa, R.F. Klosb, L.R. Willb, S.N. Gibbons-Burgenerb, A. Mvilongoa, J.Y. Abramsa, B.S. Applebyc, L.B. Schonbergera, and E.D. Belaya aNational Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (CDC), Atlanta, USA; bWisconsin Department of Health Services (WDHS), Division of Public Health, Madison, USA; cNational Prion Disease Pathology Surveillance Center (NPDPSC), Case Western Reserve University, Cleveland, USA
Aims: It is unknown whether chronic wasting disease (CWD), a prion disease of cervids, can infect people, but consumption of meat from infected animals would be the most likely route of transmission. Wisconsin Department of Health Services, Division of Public Health (WDHS) personnel maintain a database consisting of information collected from hunters who reported eating, or an intention to eat, venison from CWD-positive cervids. These data, collected since 2003, allow for the evaluation of causes of mortality in individuals potentially exposed to CWD.
Material and Methods: The WDHS database contains the name, date of birth, when available, year of CWD-positive deer harvest, and city and state of residence for each potentially exposed individual. The database also includes information on how the deer was processed (self-processed or by a commercial operator) and when applicable, names of others with whom the venison was shared. Duplicate entries (i.e., those who consumed venison from CWD-positive deer in multiple hunt years) are determined by first name, last name, and date of birth. All names in the database are cross-checked with reported cases of human prion disease in Wisconsin and cases in the National Prion Disease Pathology Surveillance Center (NPDPSC) diagnostic testing database. Persons with date of birth available are also cross-checked with prion disease decedents identified through restricted-use national multiple cause-of-death data via a data use agreement with the National Center for Health Statistics (NCHS).
Results: The database currently consists of 1561 records for hunt years 2003–2017 and 87 additional records for 2018–2019. Of these, 657 records have accompanying date of birth; 15 entries were removed as duplicates leaving 642 unique individuals. Of these individuals, 278 of 426 (66%) who ate venison from a CWD-positive deer and provided processing information reported self-processing. No matches were found among any persons in the database cross-checked with WDHS human prion disease surveillance data, NPDPSC data (February 2022 update), and NCHS data through 2020.
Conclusions: Because of the linkage of person and CWD-positive animal in the WDHS database, reviewing the cause of mortality in potentially exposed persons is possible. The number of individuals cross-checked so far is likely only a small percentage of those potentially exposed to CWD in Wisconsin, and many more years of vital status tracking are needed given an expected long incubation period should transmission to humans occur. Nevertheless, the findings of this ongoing review are thus far reassuring.
Prion disease incidence, United States, 2003–2020
R.A. Maddoxa, M.K. Persona, K. Kotobellib, A. Mvilongoa, B.S. Applebyb, L.B. Schonbergera, T.A. Hammetta, J.Y. Abramsa, and E.D. Belaya aNational Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (CDC), Atlanta, USA; bNational Prion Disease Pathology Surveillance Center (NPDPSC), Case Western Reserve University, Cleveland, USA
Aims: Mortality data, in conjunction with neuropathological and genetic testing results, are used to estimate prion disease incidence in the United States.
Material and Methods: Prion disease decedents for 2003–2020 were identified from restricted-use U.S. national multiple cause-of-death data, via a data use agreement with the National Center for Health Statistics, and from the National Prion Disease Pathology Surveillance Center (NPDPSC) database. NPDPSC decedents with neuropathological or genetic test results positive for prion disease for whom no likely match was found in the NCHS multiple cause-of-death data were added as cases for incidence calculations, while those with negative neuropathology results but with cause-of-death data indicating prion disease were removed. Unmatched cases in the NPDPSC database lacking neuropathological testing but with a positive real-time quaking-induced conversion (RT-QuIC) test result were additionally assessed. Age-specific and age-adjusted average annual incidence rates were calculated from the combined data; the year 2000 as the standard population and the direct method were used for age-adjustment.
Results: A total of 7,921 decedents were identified as having prion disease during 2003–2020 for an age-adjusted average annual incidence of 1.2 per million population. The age-adjusted incidence between males and females (1.3 and 1.1 per million, respectively) differed significantly (p < 0.0001). The age-specific average annual incidence among those <55 and ≥55 years of age was 0.2 and 4.8 per million, respectively; incidence among those ≥65 was 6.1 per million. Eighteen cases were <30 years of age for an age-specific incidence of 8.0 per billion; only 6 of these very young cases were sporadic (3 sporadic CJD, 3 sporadic fatal insomnia), with the rest being familial (9), variant (2), or iatrogenic (1). The age-adjusted annual incidence for the most recent year of data, 2020, was 1.3 per million. However, assessment of RT-QuIC positive cases lacking neuropathology in the NPDPSC database suggested that approximately 20% more cases may have occurred in that year; the addition of a subset of these cases that had date of death information available (n = 44) increased the 2020 rate to 1.4 per million.
Conclusions: Mortality data supplemented with the results of neuropathological, CSF RT-QuIC, and genetic testing can be used to estimate prion disease incidence. However, the identification in the NPDPSC database of RT-QuIC-positive cases lacking date of death information suggests that this strategy may exclude a number of probable prion disease cases. Prion disease cases <30 years of age, especially those lacking a pathogenic mutation, continue to be very rare.
Shedding of Chronic Wasting Disease Prions in Multiple Excreta Throughout Disease Course in White-tailed Deer
Nathaniel D. Denkersa, Erin E. McNultya, Caitlyn N. Krafta, Amy V. Nallsa, Joseph A. Westricha, Wilfred Goldmannb, Candace K. Mathiasona, and Edward A. Hoovera
aPrion Research Center, College of Veterinary Medicine and Biological Sciences, Department of Microbiology, Immunology, and Pathology; Colorado State University, Fort Collins, CO, USA; bDivision of Infection and Immunity, The Roslin Institute and the Royal Dick School of Veterinary Studies, University of Edinburgh, Midlothian, UK
Aims: Chronic wasting disease (CWD) now infects cervids in South Korea, North America, and Scandinavia. CWD is unique in its efficient transmission and shedding of prions in body fluids throughout long course infections. Questions remain as to the magnitude of shedding and the route of prion acquisition. As CWD continues to expand, the need to better understand these facets of disease becomes more pertinent. The purpose of the studies described was to define the longitudinal shedding profile of CWD prions in urine, saliva, and feces throughout the course of infection in white-tailed deer.
Material and Methods: Twelve (12) white-tailed deer were inoculated with either 1 mg or 300ng of CWD. Urine, saliva, and feces were collected every 3-month post-inoculation (MPI) throughout the study duration. Cohorts were established based on PNRP genotype: codon 96 GG (n = 6) and alternate codons 96 GS (n = 5) & 103NT (n = 1). Urine and saliva were analyzed using iron-oxide magnetic extraction (IOME) and real-time quaking induced conversion (RT-QuIC)(IQ). Feces were subjected to IOME, followed by 4 rounds protein misfolding cyclic amplification (PMCA) with products analyzed by RT-QuIC (IPQ). To determine whether IPQ may be superior to IQ, a subset of urine and saliva were also tested by IPQ. Results were compared with clinical disease status.
Results: Within the 96 GG cohort, positive seeding activity was detected in feces from all deer (100%), in saliva from 5 of 6 (83%), and in urine from 4 of 6 (66%). Shedding in all excreta occurred at, or just after, the first positive tonsil biopsy result. In the 96 GS/103NT cohort, positive seeding activity could be detected in feces from 3 of 6 (50%) deer, saliva in 2 of 6 (33%), and urine in 1 of 6 (16%). Shedding in excreta was detected >5 months after the first tonsil positive result. Four of six 96 GG deer developed clinical signs of CWD, whereas only 2 of the 96 GS/103NT did. Shedding was more frequently detected in deer with clinical disease. The IPQ protocol did not significantly improve detection in saliva or urine samples, however, it significantly augmented detection in feces by eliminating non-specific background commonly experienced with IQ. Negative control samples remained negative in samples tested.
Conclusions: These studies demonstrate: (a) CWD prion excretion occurs throughout infection; (2) PRNP genotype (GG≫GS/NT) influences the excreta shedding; and (3) detection sensitivity in excreta can vary with different RT-QuIC protocols. These results provide a more complete perspective of prion shedding in deer during the course of CWD infection.
Funded by: National Institutes of Health (NIH)
Grant number: RO1-NS061902-09 R to EAH, PO1-AI077774 to EAH, and R01-AI112956-06 to CKM
Acknowledgement: We abundantly thank Sallie Dahmes at WASCO and David Osborn and Gino D’Angelo at the University of Georgia Warnell School of Forestry and Natural Resources for their long-standing support of this work through provision of the hand-raised, CWD-free, white-tailed deer used in these studies
Large-scale PMCA screening of retropharyngeal lymph nodes and in white-tailed deer and comparisons with ELISA and IHC: the Texas CWD study
Rebeca Benaventea, Paulina Sotoa, Mitch Lockwoodb, and Rodrigo Moralesa
aDepartment of Neurology, McGovern Medical School, University of Texas Health Science Center at Houston, Texas, USA; bTexas Park and Wildlife Department, Texas, USA
Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy that affects various species of cervids, and both free-ranging and captive animals. Until now, CWD has been detected in 3 continents: North America, Europe, and Asia. CWD prevalence in some states may reach 30% of total animals. In Texas, the first case of CWD was reported in a free-range mule deer in Hudspeth and now it has been detected in additional 14 counties. Currently, the gold standard techniques used for CWD screening and detection are ELISA and immunohistochemistry (IHC) of obex and retropharyngeal lymph nodes (RPLN). Unfortunately, these methods are known for having a low diagnostic sensitivity. Hence, many CWD-infected animals at pre-symptomatic stages may be misdiagnosed. Two promising in vitro prion amplification techniques, including the real-time quaking-induced conversion (RT-QuIC) and the protein misfolding cyclic amplification (PMCA) have been used to diagnose CWD and other prion diseases in several tissues and bodily fluids. Considering the low cost and speed of RT-QuIC, two recent studies have communicated the potential of this technique to diagnose CWD prions in RPLN samples. Unfortunately, the data presented in these articles suggest that identification of CWD positive samples is comparable to the currently used ELISA and IHC protocols. Similar studies using the PMCA technique have not been reported.
Aims: Compare the CWD diagnostic potential of PMCA with ELISA and IHC in RPLN samples from captive and free-range white-tailed deer. Material and Methods: In this study we analyzed 1,003 RPLN from both free-ranging and captive white-tailed deer collected in Texas. Samples were interrogated with the PMCA technique for their content of CWD prions. PMCA data was compared with the results obtained through currently approved techniques.
Results: Our results show a 15-fold increase in CWD detection in free-range deer compared with ELISA. Our results unveil the presence of prion infected animals in Texas counties with no previous history of CWD. In the case of captive deer, we detected a 16% more CWD positive animals when compared with IHC. Interestingly, some of these positive samples displayed differences in their electroforetic mobilities, suggesting the presence of different prion strains within the State of Texas.
Conclusions: PMCA sensitivity is significantly higher than the current gold standards techniques IHC and ELISA and would be a good tool for rapid CWD screening.
Funded by: USDA
Grant number: AP20VSSPRS00C143
ATYPRION project: assessing the zoonotic potential of interspecies transmission of CWD isolates to livestock (preliminary results).
Enric Vidala,b, Juan Carlos Espinosac, Samanta Gilera,b, Montserrat Ordóñeza,b, Guillermo Canteroa,b, Vincent Béringued, Justin J. Greenleee, and Juan Maria Torresc
aUnitat mixta d’Investigació IRTA-UAB en Sanitat Animal. Centre de Recerca en Sanitat Animal (CReSA). Campus de la Universitat Autònoma de Barcelona (UAB), Bellaterra, Catalonia; bIRTA. Programa de Sanitat Animal. Centre de Recerca en Sanitat Animal (CReSA). Campus de la Universitat Autònoma de Barcelona (UAB), Bellaterra, Catalonia; cCentro de Investigación en Sanidad Animal, CISA-INIA-CSIC, Valdeolmos, Madrid, Spain; dMolecular Virology and Immunology, French National Research Institute for Agriculture, Food and Environment (INRAE), Université Paris-Saclay, Jouy-en-Josas, France; eVirus and Prion Research Unit, National Animal Disease Center, ARS, United States Department of Agriculture, Ames, IA, USA
Aims: Since variant Creutzfeldt-Jackob disease was linked to the consumption of bovine spongiform encephalopathy prions, the study of the pathobiological features of animal prions, particularly their zoonotic potential, is of great concern to the scientific community and public health authorities. Furthermore, interspecies transmission of prions has been demonstrated as a putative evolutionary mechanism for prions, that can lead to the emergence of new features including the ability to infect humans. For instance, small ruminants’ atypical scrapie prions, when propagated in a bovine or porcine host, can shift to a classical BSE phenotype thus posing a potential risk in case of human exposure. So far, no hard evidence of zoonotic transmission of cervids’ chronic wasting disease (CWD) to humans has been published, however experimental transmission to bovine, ovine and caprine hosts has been achieved. Our goal is to investigate if, once passaged through these domestic species, CWD prions might become infectious to humans.
Material and Methods: Different CWD isolates experimentally adapted to cattle, sheep and goat (Hamir et al, 2005, 2006, 2007, Greenlee et al 2012) have been intracerebrally inoculated to transgenic mouse models expressing the human cellular prion protein either homozygous for methionine or valine at codon 129 (Tg340-Met129 and Tg362-Val129). Additionally, inocula obtained from experimental transmission of elk CWD to ovinized (Tg501) and bovinized (BoTg110) transgenic mice, as well as white-tailed deer CWD to BoTg110 mice, are currently being bioassayed in both human PrPCtransgenic models.
Results and conclusions: No evidence of transmission has been found on first passage for bovine adapted elk and mule deer CWD to none of the humanized models. The remaining bioassays are ongoing without showing clinical signs yet, as well as second passages for the negative 1stpassages.
Funded by: La Marató de TV3 foundation. Grant number: ATYPRION (201,821–30-31-32)
PRION CONFERENCE 2022 ABSTRACTS CWD TSE PrP ZOONOSIS and ENVIRONMENTAL FACTORS
Chronic wasting disease detection in environmental and biological samples from a taxidermy site
Paulina Sotoa,b, J. Hunter Reedc, Mitch Lockwoodc, and Rodrigo Moralesa,b aDepartment of Neurology, McGovern Medical School, University of Texas Health Science Center at Houston, Texas, USA; bUniversidad Bernardo O’Higgins, Santiago, Chile; cTexas Parks and Wildlife Department, Texas, USA
Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy affecting captive and free-ranging cervids (e.g., mule deer, white-tailed deer, elk, reindeer, and moose). Nowadays, CWD is widely distributed in North America. It is suggested that CWD spreads due to direct animal contact or through exposure to contaminated environments previously inhabited by infected animals. CWD may also be spread through the movement of infected animals and carcasses. Taxidermy practices involve processing deer tissues (or whole animal carcasses). In many cases, the CWD status of processed animals is unknown. This can generate risks of disease spread and transmission. Taxidermy practices include different steps involving physical, chemical, and biological procedures. Without proper tissue handling or disposal practices, taxidermist facilities may become a focus of prion infectivity. Aims: In this study, we evaluated the presence of infectious prions in a taxidermy facility believed to be exposed to CWD. Detection was performed using the Protein Misfolding Cyclic Amplification (PMCA) technique in biological and inert environmental samples. Methods: We collected biological and environmental samples (plants, soils, insects, excreta, and others) from a taxidermy facility, and we tested these samples using the PMCA technique. In addition, we swabbed different surfaces possibly exposed to CWD-infected animals. For the PMCA reaction, we directly used a swab piece or 10 µL of 20% w/v homogenized samples. Results: The PMCA analysis demonstrated CWD seeding activity in some of the components of this facility, including insects involved in head processing, soils, and a trash dumpster. Conclusions: Different areas of this property were used for various taxidermy procedures. We were able to detect the presence of prions in i) soils that were in contact with the heads of dead animals, ii) insects involved in the cleaning of skulls, and iii) an empty dumpster where animal carcasses were previously placed. This is the first report demonstrating that swabbing is a helpful method to screen for prion infectivity on surfaces potentially contaminated with CWD. These findings are relevant as this swabbing and amplification strategy may be used to evaluate the disease status of other free-ranging and captive settings where there is a concern for CWD transmissions, such as at feeders and water troughs with CWD-exposed properties. This approach could have substantial implications for free-ranging cervid surveillance as well as in epidemiological investigations of CWD.
Funded by: USDA Grant number: AP20VSSPRS00C143
Large-scale PMCA screening of retropharyngeal lymph nodes and in white-tailed deer and comparisons with ELISA and IHC: the Texas CWD study
Rebeca Benaventea, Paulina Sotoa, Mitch Lockwoodb, and Rodrigo Moralesa aDepartment of Neurology, McGovern Medical School, University of Texas Health Science Center at Houston, Texas, USA; bTexas Park and Wildlife Department, Texas, USA
Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy that affects various species of cervids, and both free-ranging and captive animals. Until now, CWD has been detected in 3 continents: North America, Europe, and Asia. CWD prevalence in some states may reach 30% of total animals. In Texas, the first case of CWD was reported in a free-range mule deer in Hudspeth and now it has been detected in additional 14 counties. Currently, the gold standard techniques used for CWD screening and detection are ELISA and immunohistochemistry (IHC) of obex and retropharyngeal lymph nodes (RPLN). Unfortunately, these methods are known for having a low diagnostic sensitivity. Hence, many CWD-infected animals at pre-symptomatic stages may be misdiagnosed. Two promising in vitro prion amplification techniques, including the real-time quaking-induced conversion (RT-QuIC) and the protein misfolding cyclic amplification (PMCA) have been used to diagnose CWD and other prion diseases in several tissues and bodily fluids. Considering the low cost and speed of RT-QuIC, two recent studies have communicated the potential of this technique to diagnose CWD prions in RPLN samples. Unfortunately, the data presented in these articles suggest that identification of CWD positive samples is comparable to the currently used ELISA and IHC protocols. Similar studies using the PMCA technique have not been reported. Aims: Compare the CWD diagnostic potential of PMCA with ELISA and IHC in RPLN samples from captive and free-range white-tailed deer. Material and Methods: In this study we analyzed 1,003 RPLN from both free-ranging and captive white-tailed deer collected in Texas. Samples were interrogated with the PMCA technique for their content of CWD prions. PMCA data was compared with the results obtained through currently approved techniques. Results: Our results show a 15-fold increase in CWD detection in free-range deer compared with ELISA. Our results unveil the presence of prion infected animals in Texas counties with no previous history of CWD. In the case of captive deer, we detected a 16% more CWD positive animals when compared with IHC. Interestingly, some of these positive samples displayed differences in their electroforetic mobilities, suggesting the presence of different prion strains within the State of Texas. Conclusions: PMCA sensitivity is significantly higher than the current gold standards techniques IHC and ELISA and would be a good tool for rapid CWD screening.
Funded by: USDA Grant number: AP20VSSPRS00C143
Protein misfolding cyclic amplification (PMCA) as an ultra-sensitive technique for the screening of CWD prions in different sample types
Francisca Bravo‐Risia,b, Paulina Sotoa,b, Rebeca Benaventea, Hunter Reedc, Mitch Lockwoodc, Tracy Nicholsd, and Rodrigo Moralesa,b aDepartment of Neurology, The University of Texas Health Science Center at Houston, Houston, TX, USA; bCentro Integrativo de Biologia y Quimica Aplicada (CIBQA), Universidad Bernardo O’Higgins, Santiago, Chile; cTexas Park and Wildlife Department, Texas, USA; dVeterinary Services Cervid Health Program, United States Department of Agriculture, Animal and Plant Health Inspection Service, Fort Collins, Colorado, USA
Chronic wasting disease (CWD) is a prion disease that affects farmed and free-ranging cervids. The infectious agent in CWD is a misfolded form of the prion protein (PrPSc) that promotes conformational changes in the host’s cellular prion protein (PrPC). Currently, definitive CWD status is confirmed in the brain and lymphoid tissues by immunohistochemistry. The limitation of this technique is its poor sensitivity. Protein misfolding cyclic amplification (PMCA) and real-time quaking-induced conversion (RT- QuIC) are ultra-sensitive techniques that overcome these issues. PMCA mimics the self- propagation of infectious prions in vitro through multiple incubation/sonication cycles, increasing the number of prion particles present in a given sample. The detection of proteinase K (PK) -resistant PrPScby PMCA has been performed in experimental and natural samples that might harbor subclinical levels of prions. These samples include several tissues, bodily fluids, excreta, and different manmade and natural materials, including mineral licks, soils, and plants. Aims: In this study, we highlight recent advances and contributions that our group has performed in the detection of CWD prions from samples collected in farmed and free-ranging cervids, as well as other specimens involving the environment that contains CWD-infected deer. Material and Methods: A set of diverse samples analyzed in this study were collected by USDA and TPWD personnel in breeding and taxidermy facilities, and deer breeding facilities. These included animal and environmental samples. Additional samples from free-ranging animals were provided by hunters. Results: The diverse range of samples successfully detected for CWD prion infection in this study include blood, semen, feces, obex, retropharyngeal lymph node, fetuses (neural and peripheral tissues) and gestational tissues, parasites, insects, plants, compost/soil mixtures, and swabs from trash containers. Importantly, these results helped to identify seeding-competent prions in places reported to be free of CWD. The levels of prion infectivity in most of these samples are currently being investigated. Conclusions: Our findings contribute to the understanding of the transmission dynamics and prevalence of CWD. In addition, our data have helped to identify CWD in areas previously considered to be free of CWD. We also demonstrate that PMCA is a powerful technique for the screening of biological and environmental samples. Overall, our research suggests that PMCA may be a useful tool to implement for the surveillance and management of CWD. Funded by: NIH/NIAID and USDA Grant number: 1R01AI132695 (NIH) and AP20VSSPRS00C143 (USDA)
Nasal bot: an emerging vector for natural chronic wasting disease transmission
Paulina Sotoa,b, Francisca Bravo-Risia,b, Carlos Kramma, Nelson Pereza, Rebeca Benaventea, J. Hunter Reedc, Mitch Lockwoodc, Tracy A. Nicholsd, and Rodrigo Moralesa,b aDepartment of Neurology, McGovern Medical School, University of Texas Health Science Center at Houston, Texas, USA; bUniversidad Bernardo O’Higgins, Santiago, Chile; cTexas Park and Wildlife Department, Texas, USA; dVeterinary Services Cervid Health Program, United States Department of Agriculture, Animal and Plant Health Inspection Service, Fort Collins, Colorado, USA
Chronic wasting disease (CWD) is a fatal neurodegenerative disease that affects farmed and free-ranging cervids populations. The spread of CWD in cervids is thought to occur through the direct contact between cervids or through the exposure of naïve animals to contaminated environments. Parasites are known vectors of multiple diseases in animals. However, the potential role of parasites in CWD transmission remains unclear. Aims: The main objective of this study was to determine if CWD prions could be detected in the larvae of deer nasal bot flies, a common deer parasite, taken from CWD-infected white-tailed deer (Odocoileus virginianus). Methods: Bot fly larvae were collected from the nasal cavity of naturally infected CWD- positive or CWD non-detect white-tailed deer. The CWD seeding activity of the larvae was interrogated by PMCA. Prion infectivity was also evaluated in cervidized transgenic mouse bioassay (intra-cerebral administration in Tg1536 mice). Mice inoculated with bot larvae homogenate were sacrificed when they showed established signs of prion disease, or at extended periods after treatment (600 days). All inoculated mouse brains were evaluated for protease resistant prions to confirm clinical or sub-clinical infection. Bot larvae from CWD non-detect deer were used as controls. To further mimic environmental transmission, bot larvae homogenates were mixed with soils and plants were grown on them. Both plants and soils were tested for prion seeding activity. Results: PMCA analysis demonstrated CWD seeding activity in nasal bot larvae from captive and free-ranging white-tailed deer. CWD-contaminated bots efficiently infected transgenic mice, with attack rates and incubation periods suggesting high infectivity titers. Further analyses of treated animals (biochemical characterization of protease resistant prions and immunohistochemistry) confirmed prion infection. Analyses on dissected parts of the bot larvae demonstrate that the infectivity is concentrated in the larvae cuticle (outer part). Nasal bot larvae extracts mixed with
soils showed seeding activity by PMCA. Interestingly, plants grown in soil contaminated with the nasal bot larvae extract were found to produce seeding activity by PMCA. Conclusion: In this study we described for the first time that deer nasal bot larvae from CWD-infected deer carry high CWD infectivity titers. We also demonstrate that CWD prions in these parasites can interact with other environmental components relevant for disease transmission. Considering this information, we propose that deer nasal bot larvae could act as vectors for CWD transmission in wild and farming settings. Funded by: NIH/NIAID and USDA/APHIS Grant number: R01AI132695 and AP20VSSPRS00C143 PRION 2022 ABSTRACTS, AND A BIG THANK YOU TO On behalf of the Prion2020/2022 Congress Organizing Committee and the NeuroPrion Association, we heartily invite you to join us for the International Conference Prion2020/2022 from 13.-16. September 2022 in Göttingen.
Prion 2022 Conference abstracts: pushing the boundaries
Shedding of Chronic Wasting Disease Prions in Multiple Excreta Throughout Disease Course in White-tailed Deer
Nathaniel D. Denkersa, Erin E. McNultya, Caitlyn N. Krafta, Amy V. Nallsa, Joseph A. Westricha, Wilfred Goldmannb, Candace K. Mathiasona, and Edward A. Hoovera
aPrion Research Center, College of Veterinary Medicine and Biological Sciences, Department of Microbiology, Immunology, and Pathology; Colorado State University, Fort Collins, CO, USA; bDivision of Infection and Immunity, The Roslin Institute and the Royal Dick School of Veterinary Studies, University of Edinburgh, Midlothian, UK
Aims: Chronic wasting disease (CWD) now infects cervids in South Korea, North America, and Scandinavia. CWD is unique in its efficient transmission and shedding of prions in body fluids throughout long course infections. Questions remain as to the magnitude of shedding and the route of prion acquisition. As CWD continues to expand, the need to better understand these facets of disease becomes more pertinent. The purpose of the studies described was to define the longitudinal shedding profile of CWD prions in urine, saliva, and feces throughout the course of infection in white-tailed deer.
Material and Methods: Twelve (12) white-tailed deer were inoculated with either 1 mg or 300ng of CWD. Urine, saliva, and feces were collected every 3-month post-inoculation (MPI) throughout the study duration. Cohorts were established based on PNRP genotype: codon 96 GG (n = 6) and alternate codons 96 GS (n = 5) & 103NT (n = 1). Urine and saliva were analyzed using iron-oxide magnetic extraction (IOME) and real-time quaking induced conversion (RT-QuIC)(IQ). Feces were subjected to IOME, followed by 4 rounds protein misfolding cyclic amplification (PMCA) with products analyzed by RT-QuIC (IPQ). To determine whether IPQ may be superior to IQ, a subset of urine and saliva were also tested by IPQ. Results were compared with clinical disease status.
Results: Within the 96 GG cohort, positive seeding activity was detected in feces from all deer (100%), in saliva from 5 of 6 (83%), and in urine from 4 of 6 (66%). Shedding in all excreta occurred at, or just after, the first positive tonsil biopsy result. In the 96 GS/103NT cohort, positive seeding activity could be detected in feces from 3 of 6 (50%) deer, saliva in 2 of 6 (33%), and urine in 1 of 6 (16%). Shedding in excreta was detected >5 months after the first tonsil positive result. Four of six 96 GG deer developed clinical signs of CWD, whereas only 2 of the 96 GS/103NT did. Shedding was more frequently detected in deer with clinical disease. The IPQ protocol did not significantly improve detection in saliva or urine samples, however, it significantly augmented detection in feces by eliminating non-specific background commonly experienced with IQ. Negative control samples remained negative in samples tested.
Conclusions: These studies demonstrate: (a) CWD prion excretion occurs throughout infection; (2) PRNP genotype (GG≫GS/NT) influences the excreta shedding; and (3) detection sensitivity in excreta can vary with different RT-QuIC protocols. These results provide a more complete perspective of prion shedding in deer during the course of CWD infection.
Funded by: National Institutes of Health (NIH)
Grant number: RO1-NS061902-09 R to EAH, PO1-AI077774 to EAH, and R01-AI112956-06 to CKM
Acknowledgement: We abundantly thank Sallie Dahmes at WASCO and David Osborn and Gino D’Angelo at the University of Georgia Warnell School of Forestry and Natural Resources for their long-standing support of this work through provision of the hand-raised, CWD-free, white-tailed deer used in these studies
Carrot plants as potential vectors for CWD transmission
Paulina Sotoa,b, Francisca Bravo-Risia,b, Claudio Sotoa, and Rodrigo Moralesa,b
aDepartment of Neurology, McGovern Medical School, University of Texas Health Science Center at Houston, Texas, USA; bUniversidad Bernardo O’Higgins, Santiago, Chile
Prion diseases are infectious neurodegenerative disorders afflicting humans and other mammals. These diseases are generated by the misfolding of the cellular prion protein into a disease-causing isoform. Chronic wasting disease (CWD) is a prevalent prion disease affecting cervids (captive and free-range). CWD is thought to be transmitted through direct animal contact or by indirect exposure to contaminated environments. Many studies have shown that infectious prions can enter the environment through saliva, feces, or urine from infected animals and decaying carcasses. However, we do not fully understand the specific contribution of each component to disease transmission events. Plants are logical environmental components to be evaluated since they grow in environments contaminated with CWD prions and are relevant for animal and human nutrition.
Aims: The main objective of this study is to study whether prions are transported to the roots and leaves of carrots, an edible plant commonly used in the human diet and as deer bait.
Methods: We have grown carrot plants in CWD-infected soils. After 90 days, we harvested the carrots and separated them from the leaves. The experiment was controlled by growing plants in soil samples treated with brain extracts from healthy animals. These materials were interrogated for their prion seeding activity using the Protein Misfolding Cyclic Amplification (PMCA) technique. Infectivity was evaluated in mouse bioassays (intracerebral injections in Tg1536 mice). The animals were sacrificed when they showed established signs of prion disease. Animals not displaying clinical signs were sacrificed at 600 days post-inoculation.
Results: The PMCA analysis demonstrated CWD seeding activity in soils contaminated with CWD prions, as well as in carrot plants (leaves and roots) grown on them. Bioassays demonstrated that both leaves and roots contained CWD prions in sufficient quantities to induce disease (92% attack rate). As expected, animals treated with prion-infected soils developed prion disease at shorter incubation periods (and complete attack rates) compared to plant components. Animals treated with soil and plant components exposed with CWD-free brain extracts did not display prion-associated clinical signs or evidence of sub-clinical prion infection.
Conclusions: We show that edible plant components can absorb prions from CWD contaminated soils and transport them to their aerial parts. Our results indicate that plants could participate as vectors of CWD transmission. Importantly, plants designated for human consumption represent a risk of introducing CWD prions into the human food chain.
Funded by: NIH
Grant number: R01AI132695
***Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility.***
Even if the prevailing view is that sporadic CJD is due to the spontaneous formation of CJD prions, it remains possible that its apparent sporadic nature may, at least in part, result from our limited capacity to identify an environmental origin.
O.05: Transmission of prions to primates after extended silent incubation periods: Implications for BSE and scrapie risk assessment in human populations
Emmanuel Comoy, Jacqueline Mikol, Valerie Durand, Sophie Luccantoni, Evelyne Correia, Nathalie Lescoutra, Capucine Dehen, and Jean-Philippe Deslys Atomic Energy Commission; Fontenay-aux-Roses, France
Prion diseases (PD) are the unique neurodegenerative proteinopathies reputed to be transmissible under field conditions since decades. The transmission of Bovine Spongiform Encephalopathy (BSE) to humans evidenced that an animal PD might be zoonotic under appropriate conditions. Contrarily, in the absence of obvious (epidemiological or experimental) elements supporting a transmission or genetic predispositions, PD, like the other proteinopathies, are reputed to occur spontaneously (atpical animal prion strains, sporadic CJD summing 80% of human prion cases).
Non-human primate models provided the first evidences supporting the transmissibiity of human prion strains and the zoonotic potential of BSE. Among them, cynomolgus macaques brought major information for BSE risk assessment for human health (Chen, 2014), according to their phylogenetic proximity to humans and extended lifetime. We used this model to assess the zoonotic potential of other animal PD from bovine, ovine and cervid origins even after very long silent incubation periods.
*** We recently observed the direct transmission of a natural classical scrapie isolate to macaque after a 10-year silent incubation period,
***with features similar to some reported for human cases of sporadic CJD, albeit requiring fourfold long incubation than BSE. Scrapie, as recently evoked in humanized mice (Cassard, 2014),
***is the third potentially zoonotic PD (with BSE and L-type BSE),
***thus questioning the origin of human sporadic cases.
We will present an updated panorama of our different transmission studies and discuss the implications of such extended incubation periods on risk assessment of animal PD for human health.
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***thus questioning the origin of human sporadic cases***
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***our findings suggest that possible transmission risk of H-type BSE to sheep and human. Bioassay will be required to determine whether the PMCA products are infectious to these animals.
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PRION 2015 CONFERENCE
***Transmission data also revealed that several scrapie prions propagate in HuPrP-Tg mice with efficiency comparable to that of cattle BSE. While the efficiency of transmission at primary passage was low, subsequent passages resulted in a highly virulent prion disease in both Met129 and Val129 mice.
***Transmission of the different scrapie isolates in these mice leads to the emergence of prion strain phenotypes that showed similar characteristics to those displayed by MM1 or VV2 sCJD prion.
***These results demonstrate that scrapie prions have a zoonotic potential and raise new questions about the possible link between animal and human prions.
PRION 2016 TOKYO
Saturday, April 23, 2016
SCRAPIE WS-01: Prion diseases in animals and zoonotic potential 2016
Prion. 10:S15-S21. 2016 ISSN: 1933-6896 printl 1933-690X online
Taylor & Francis
Prion 2016 Animal Prion Disease Workshop Abstracts
WS-01: Prion diseases in animals and zoonotic potential
Transmission of the different scrapie isolates in these mice leads to the emergence of prion strain phenotypes that showed similar characteristics to those displayed by MM1 or VV2 sCJD prion.
These results demonstrate that scrapie prions have a zoonotic potential and raise new questions about the possible link between animal and human prions.
Title: Transmission of scrapie prions to primate after an extended silent incubation period)
*** In complement to the recent demonstration that humanized mice are susceptible to scrapie, we report here the first observation of direct transmission of a natural classical scrapie isolate to a macaque after a 10-year incubation period. Neuropathologic examination revealed all of the features of a prion disease: spongiform change, neuronal loss, and accumulation of PrPres throughout the CNS.
*** This observation strengthens the questioning of the harmlessness of scrapie to humans, at a time when protective measures for human and animal health are being dismantled and reduced as c-BSE is considered controlled and being eradicated.
*** Our results underscore the importance of precautionary and protective measures and the necessity for long-term experimental transmission studies to assess the zoonotic potential of other animal prion strains.
Research Project: Pathobiology, Genetics, and Detection of Transmissible Spongiform Encephalopathies
Location: Virus and Prion Research
Title: Transmission of the atypical/nor98 scrapie agent to suffolk sheep with VRQ/ARQ, ARQ/ARQ, and ARQ/ARR genotypes
Author item Cassmann, Eric item MAMMADOVA, JAJIBA - Orise Fellow item BENESTAD, SYLVIE - Norwegian Veterinary Institute item MOORE, SARA JO - Orise Fellow item Greenlee, Justin
Submitted to: PLoS ONE Publication Type: Peer Reviewed Journal Publication Acceptance Date: 1/21/2021 Publication Date: 2/11/2021
Citation: Cassmann, E.D., Mammadova, J., Benestad, S., Moore, S., Greenlee, J.J. 2021. Transmission of the atypical/nor98 scrapie agent to suffolk sheep with VRQ/ARQ, ARQ/ARQ, and ARQ/ARR genotypes. PLoS ONE. 16(2). Article e0246503. https://doi.org/10.1371/journal.pone.0246503. DOI: https://doi.org/10.1371/journal.pone.0246503
Interpretive Summary: Atypical scrapie is a prion disease that affects sheep. Unlike classical scrapie, atypical scrapie is thought to occur spontaneously, and it is unlikely to transmit between sheep under natural conditions. Another notable distinction between classical and atypical scrapie is the prion protein genotype of afflicted sheep and the locations in the brain where misfolded prions accumulate. Atypical scrapie generally occurs in sheep that are resistant to classical scrapie. Misfolded prions are predominantly found in the cerebellum for atypical scrapie and not in the brainstem as seen with classical scrapie. Atypical scrapie is a relevant disease because of its potential association with other prion diseases. Some research has shown that the atypical scrapie agent can undergo a transformation of disease forms that makes it appear like classical scrapie or classical bovine spongiform encephalopathy (mad cow disease). Therefore, atypical scrapie is thought to be a possible source for these prion diseases. We investigated the transmission of the atypical scrapie agent to sheep with three different prion protein genotypes. A diagnosis of atypical scrapie was made in all three genotypes of sheep. Misfolded prion protein was detected earliest in the cerebellum and the retina. This is the first report describing the early accumulation of misfolded prions in the retina of sheep with atypical scrapie. Understanding where misfolded prions accumulate in cases of atypical scrapie can lead to better detection earlier in the disease. Furthermore, the materials derived from this experiment will aid in investigating origins of other prion diseases.
Technical Abstract: Scrapie is a transmissible spongiform encephalopathy that occurs in sheep. Atypical/Nor98 scrapie occurs in sheep with that tend to be resistant to classical scrapie and it is thought to occur spontaneously. The purpose of this study was to test the transmission of the Atypical/Nor98 scrapie agent in three genotypes of Suffolk sheep and characterize the distribution of misfolded prion protein (PrPSc). Ten sheep were intracranially inoculated with brain homogenate from a sheep with Atypical/Nor98 scrapie. All sheep with the ARQ/ARQ and ARQ/ARR genotypes developed Atypical/Nor98 scrapie confirmed by immunohistochemistry, and one (1/3) sheep with the VRQ/ARQ genotype had detectable PrPSc consistent with Atypical/Nor98 scrapie at the experimental endpoint of 8 years. Sheep with mild early accumulations of PrPSc in the cerebellum had concomitant retinal PrPSc. Accordingly, large amounts of retinal PrPSc were identified in clinically affected sheep and sheep with dense accumulations of PrPSc in the cerebellum.
Research Project: Pathobiology, Genetics, and Detection of Transmissible Spongiform Encephalopathies Location: Virus and Prion Research
Title: Scrapie in white-tailed deer: a strain of the CWD agent that efficiently transmits to sheep?
Author item Greenlee, Justin item KOKEMULLER, ROBYN - US Department Of Agriculture (USDA) item MOORE, S - Oak Ridge Institute For Science And Education (ORISE) item WEST GREENLEE, M - Iowa State University
Submitted to: Meeting Abstract Publication Type: Abstract Only Publication Acceptance Date: 3/29/2019 Publication Date: N/A Citation: N/A
Interpretive Summary:
Technical Abstract: Scrapie is a transmissible spongiform encephalopathy of sheep and goats that is associated with widespread accumulation of abnormal prion protein (PrPSc) in the central nervous and lymphoid tissues. Chronic wasting disease (CWD) is the natural prion disease of cervid species, and the tissue distribution of PrPSc in affected cervids is similar to scrapie in sheep. There are several lines of evidence that suggest that multiple strains of CWD exist, which may affect the agent’s potential to transmit to hosts of the same or different species. We inoculated white-tailed deer with the scrapie agent from ARQ/ARQ sheep, which resulted in 100% attack rates by either the intracranial or oronasal route of inoculation. When examining tissues from the brainstems or lymphoid tissues by traditional diagnostic methods such as immunohistochemistry or western blots, it is difficult to differentiate tissues from deer infected with scrapie from those infected with CWD. However, there are several important differences between tissues from scrapie-infected white-tailed deer (WTD scrapie) and those infected with CWD (WTD CWD). First, there are different patterns of PrPSc deposition in the brains of infected deer: brain tissues from deer with WTD scrapie had predominantly particulate and stellate immunoreactivity whereas those from deer with WTD-CWD had large aggregates and plaque-like staining. Secondly, the incubation periods of WTD scrapie isolates are longer than CWD isolates in mice expressing cervid prion protein. Most notably, the transmission potential of these two isolates back to sheep is distinctly different. Attempts to transmit various CWD isolates to sheep by the oral or oronasal routes have been unsuccessful despite observation periods of up to 7 years. However, WTD scrapie efficiently transmitted back to sheep by the oronasal route. Upon transmission back to sheep, the WTD scrapie isolate exhibited different phenotypic properties when compared to the sheep receiving the original sheep scrapie inoculum including different genotype susceptibilities, distinct PrPSc deposition patterns, and much more rapid incubation periods in transgenic mice expressing the ovine prion protein. The scrapie agent readily transmits between sheep and deer after oronasal exposure. This could confound the identication of CWD strains in deer and the eradication of scrapie from sheep.
''The scrapie agent readily transmits between sheep and deer after oronasal exposure. This could confound the identication of CWD strains in deer and the eradication of scrapie from sheep.''
RT-QuIC detection of pathological prion protein in subclinical goats following experimental oral transmission of L-type BSE
Alessandra Favole1* , Maria Mazza1 , Antonio D’Angelo2 , Guerino Lombardi3 , Claudia Palmitessa1 , Luana Dell’Atti1 , Giulia Cagnotti2 , Elena Berrone1 , Marina Gallo1 , Tiziana Avanzato1 , Erika Messana1 , Loretta Masoero1 , Pier Luigi Acutis1 , Daniela Meloni1 , Franco Cardone4 , Maria Caramelli1 , Cristina Casalone1 and Cristiano Corona1*
Abstract
Objective: The spread of bovine spongiform encephalopathy (BSE) agent to small ruminants is still a major issue in the surveillance of transmissible spongiform encephalopathies (TSEs). L-type bovine spongiform encephalopathy (L-BSE) is an atypical form of BSE with an unknown zoonotic potential that is transmissible to cattle and small ruminants. Our current knowledge of bovine atypical prion strains in sheep and goat relies only on experimental transmission studies by intracranial inoculation. To assess oral susceptibility of goats to L-BSE, we orally inoculated five goats with cattle L-BSE brain homogenates and investigated pathogenic prion protein (PrPsc) distribution by an ultrasensitive in vitro conversion assay known as Real-Time Quaking Induced Conversion (RT-QuIC).
Results: Despite a prolonged observation period of 80 months, all these animals and the uninfected controls did not develop clinical signs referable to TSEs and tested negative by standard diagnostics. Otherwise, RT-QuIC analysis showed seeding activity in five out of five examined brain samples. PrPsc accumulation was also detected in spinal cord and lymphoreticular system. These results indicate that caprine species are susceptible to L-BSE by oral transmission and that ultrasensitive prion tests deserve consideration to improve the potential of current surveillance systems against otherwise undetectable forms of animal prion infections.
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Discussion and conclusions
Data here presented indicate that caprine species are susceptible to L-BSE after oral administration and are able to produce very low levels of prions in both lymphatic and central nervous tissues as demonstrated by optimized, high-sensitive, RT-QuIC assay.
At variance with goats intracerebrally infected with L-BSE [4], in this study, no animal developed clinical signs of disease despite prolonged periods of observation, suggesting a comparatively low efficiency of the oral route versus the intracerebral one in L-BSE, a feature that further distinguish this strain from classical BSE [14, 15].
Interestingly, all goats tested negative by standard diagnostics for PrPsc performed on brainstem. This finding, associated with the low amount of PrPsc detected in different brain areas, suggests a partial strain-specific transmission barrier. Indeed, inoculation of a prion into a new host species can produce prolonged incubation periods and/or subclinical infection [16, 17]. In addition, the lack of clinical signs suggests that naturally L-BSE-infected goats may be asymptomatic similarly to what proposed by Okada et al. for oral L-BSE in cattle [17].
In line with previous results [18], RT-QuIC detected lower levels of prions than traditional diagnostic tools. Rapid and confirmatory tests failed to identify any PrPsc in the subclinical animals, while RT-QuIC allowed us to detect misfolded prion protein in multiple brain regions, spinal cord and lymphoreticular system. Studies have established that the rate of fluorescence increase in RTQuIC, while not measuring infectivity, is directly related to the concentration of prions in the sample seeding the reaction [19, 20]. Prolonged lag phases of RT-QuIC reactions indicate relatively low amounts of PrPsc in the examined tissues and may reassure about the possibility of goat to play as silent L-BSE spreaders in natural conditions. However, we believe that prudence must be always adopted when dealing with the risk of prion spread in field conditions as also suggested by recent data by Denkers and colleagues, who showed that the oral route of infection for chronic wasting disease in deer, may be much more efficient than previously thought [21]. Furthermore, although the mere presence of PrPsc is not indicative of a possible infectivity of the tissue, the finding of positivity in the lymphoreticular tissue must alert to the potential distribution of PrPsc in peripheral body regions which may increase the risks for humans. Bioassay of infectivity by inoculation of susceptible animals with brains of these goats may help to clarify this issue.
Based on the results achieved with this prion form and also other animal strains, it would be useful to consider the possibility to enlarge current diagnostic criteria to include, in defined conditions (e.g. very limited amounts of source tissue, or preclinical testing), the application of ultrasensitive diagnostic methods. This will not only improve the sensitivity of our surveillance systems but will also help to protect food chain from accidental spillovers of the agent of L-BSE.
Limitations
Te primary limitation of this work is that infectivity was not demonstrated by bioassay and the infectious titre was not determined. Terefore, we cannot comment the degree of risk for human.
Despite these limitations, this work specifcally demonstrates prion-seeding activity in tissues of goats orally exposed to L-BSE and provide RT-QuIC as useful method to enhance surveillance of TSEs.
Keywords: Prion, L-BSE, RT-QuIC, Goat, Oral transmission, PrPsc, Ultrasensitive detection
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TUESDAY, NOVEMBER 29, 2022
CHRONIC WASTING DISEASE DETECTION AND MANAGEMENT: WHAT HAS WORKED AND WHAT HAS NOT?
Control of Chronic Wasting Disease OMB Control Number: 0579-0189 APHIS-2021-0004 Singeltary Submission
Docket No. APHIS-2018-0011 Chronic Wasting Disease Herd Certification
APHIS Indemnity Regulations [Docket No. APHIS-2021-0010] RIN 0579-AE65 Singeltary Comment SubmissionComment from Singeltary Sr., Terry
Posted by the Animal and Plant Health Inspection Service on Sep 8, 2022
Scrapie Field Trial was developed at Mission, Texas, on 450 acres of pastureland, part of the former Moore Air Force Base
EPIDEMIOLOGY OF SCRAPIE IN THE UNITED STATES
Academic Preg
James Hourriganl, Albert Klingsporn2, Edited by » Peast
W. W. Clark3, and M, de Camp4
United States Department of Agriculture, Animal and Plant Health Inspection Service, Veterinary Services
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METHODS
A Scrapie Field Trial was developed at Mission, Texas, to provide additional information for the eradication program on the epidemiology of natural scrapie. The Mission Field Trial Station is located on 450 acres of pastureland, part of the former Moore Air Force Base, near Mission,
Texas. It was designed to bring previously exposed, and later also unexposed, sheep or goats to the Station and maintain and breed them under close observation for extended periods
to determine which animals would develop scrapie and define more closely the natural spread and other epidemiological aspects of the disease.
The 547 previously exposed sheep brought to the Mission Station beginning in 1964 were of the Cheviot, Hampshire, Montadale, or Suffolk breeds. They were purchased as field outbreaks occurred, and represented 21 bloodlines in which scrapie had been diagnosed. Upon arrival at the Station, the sheep were maintained on pasture, with supplemental feeding as necessary. The station was divided into 2 areas:
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RESULTS
Table 1 indicated that previously exposed sheep brought to the Station at various times and ages (1 to 89 months old) included 333 Suffolks at risk. Of these, 98 (29%) developed scrapie. This demonstrated the necessity to slaughter such sheep to prevent further Spread of the disease, These pre- viously exposed Suffolks were bred at the Station and produced 446 progeny at risk. Of these 153 (34%) developed scrapie.
Although the minimum and average ages when scnapied were similar for both groups, some of the previously exposed Suffolks brought to the Station developed scrapie when much older--ewes 60 to 142 months old and rams 67 to 102 months old. O£ the 153 Suffolks born at the Station, only 3 were more than 60 months of age (65, 66, and 69 months old).
This difference in age scrapied was attributed to the fact that the Suffolks born at the Station may have been sub- ject to a greater exposure from birth.
It was also observed that when both dam and progeny were scrapied, the progeny nearly always developed clinical disease at a younger age than their respective dam. Thirty- two dams were scrapied at an average of 60 months of age. Forty-six of their progeny developed the disease at an average of 38 months (range 25 to 53 months). Thirty-seven of the 46 progeny were younger than the dam (average 20 months younger, range 2 to 99 months younger). Two were scrapied at the same age as their dams, and 8 were older (average 5 months, range 1 to 13 months older).
++. Although the incidence of scrapie was considerably Greater in the progeny of scrapied compared to free dams, the progeny of either scrapied or free dams manifested scrapie at the typical age and irrespective of the age their respective dams were scrapied. The differences in ages that dams and progeny were scrapied was believed due to difference of exposure, particularly whether they were exposed at an early age,
Table 2 summarized the data on exposed Suffolks and was Prepared so as to show scrapie incidence in the progeny of dams and sires of known Scrapie status. The scrapie incidence in the progeny of Free X Free parents was 25%, progeny of scrapied Sires 39%, and scrapied dams 42%. When both sire and dam were scrapied, the scrapie incidence in 18 Progeny at risk was 78%.
When the scrapie status of the sire was ignored, scrapie incidence in th- progeny of free dams was 34% and in pre y of scrapied da as 62%. When the scrapie status of the dam was ignored, scrapie incidence in the progeny of free sires was 26% and in the progeny of scrapied sires was 452.
Although the scrapie incidence was nearly double in the progeny of scrapied compared to free dams, the latter con- tributed a greater number of scrapied progeny, 116, compared to only 51 cases which had scrapied dams. This was because free dams made a considerably heavier contribution to the progeny at risk4-342 compared to 82. It was felt that in farm flocks a similar situation could exist.
It was possible that free dams could have been mis- classified; however, this was unlikely to have been significant, unless "nonclinical or carrier" dams exist. In this Suffolk group, the ages of 100 free dams of scrapied progeny ranged from 25 to 160 (average 97) months. These free dams did not show clinical signs of scrapie,”and there were no histopathological lesions suggesting scrapie in those which died, If one cannot classify as free, ewes which have reached 97 months (average) and did not develop the disease, from a practical standpoint, it is not possible to classify sheep as free, at least on the basis of clinical signs and histology. The free dams of 50% of the scrapied progeny were more than 100 months of age, averaging 126 months.
Upon arrival at the Mission Station at 3 to 9 months of age, the 140 previously unexposed sheep and goats were placed in infected pastures and corrals and were subjected to con- tact with a succession of natural cases of scrapie in sheep, and eventually also in goats. These animals were bred only within their respective groups and were not crossbred to other breeds of sheep or those brought to the Station from infected flocks or their progeny. The male or female animals mixed freely with animals of their respective sex of the infected Flock and were similarly identified and subjected to similar flock management and diagnostic procedures.
Table 3 indicated that natural scrapie had occurred in 5 of the 140 previously unexposed sheep. One case each occurred in Rambouillet, Targhee, and Hampshire ewes at 88, 89, and 89 months of age and in % Suffolk ewes at 73 and 102 months of age, and 85, 82, 80, 64, and 93 months following initial natural exposure. This represented a natural situation involving lateral spread, under the circumstances involved, when sheep were not exposed when very young. Scrapie was not detected clinicaliy or histologically in any of the dairy or Angora goats brought to the Station. The disease occurred in an average of 27% of the progeny of previously unexposed sheep or goats born at the Station and included cases in progeny of all breeds of sheep or goats taken there, The incidence in the progeny ranged from 14% in Rambouillet sheep to 61% in dairy goats. ~
These data showed that scrapie spread laterally, by contact exposure, from scrapied te previously free animals, but at an apparently lower rate when exposure was first received at the age of 3 to 9 months. These animals were presumed to be susceptible to the disease, as their progeny developed scrapie at rates and ages similar (on the average) to the progeny, pf previously exposed Suffolk sheep born and reared in the same environment.
It was suggested that the progeny of previously unexposed animals developed scrapie at a much higher rate than their parents, and at a younger age, because they were subjected to exposure from birth. The data did not rule out the possibility that the animals born at the Station could have also received the virus from their dams "vertically" prior te, at, or following birth.
Table 4 summarized the scrapie incidence in #he progeny, born at the Station, of previously unexposed dairy goats.
The data were prepared so as to show scrapie incidence in the progeny of dams and sires of known scrapie status.
The 58% incidence in the progeny (24 at risk) of Free X Free parents was more than twice the 25% seen in the Suffolk group (Table 2). Scrapied sires did not increase the incidence in goat progeny (it was 44%); scrapied dams increased the incidence to 71%. When both sire and dam were scrapied the incidence was 89%, with only 9 goat progeny at risk.
When the scrapie status of the sire was ignored, the scrapie incidence in the progeny of free dams was 56% and in the progeny of scrapied dams it was 74%.
Free dams contributed 34 progeny at risk and scrapied dams 31 progeny.
When the scrapie status of the dam was ignored, scrapie incidence was 64% in the progeny of free sires and a similar 66% in the progeny of scrapied sires.
A total of 244 sheep (127 Suffolk, 59 Rambouillet, and 58 Targhee) were removed from scrapie exposure within a few hours of birth or at 4, 9, or 20 months of age and placed in isolation pens. Removal of sheep from exposure at these ages was selected as being representative of usual flock operations when sheep might be sold from an infected flock at weaning, the first fall or the second fall after their birth.
Table 5 reflected the fate of such animals. Four of the 6 scrapied sheep which had been isolated at birth were Suffolks and the 2 older animals were Targhees. The first case in the group isolated at birth was a Targhee, progeny of a ewe that did not develop clinical scrapie. The scrapie incidence in 36 at risk Suffolks removed from exposure at birth was 11%, con- siderably less -“en that expected had these animals remz d in an infected en ment.
Table 6 reflected the status of 51 goats isolated from scrapie exposure at birth, and at 6, 8 to 10, 20, 32 to 59 and 60 to 82 months of age.
None of the goats removed at birth developed scrapie, although all 5 of those alive at 5 years of age had scrapied dams and 1 also had a scrapied sire. The sire of the remaining 4 had sired 7 scrapied progeny. Under such circumstances, had they remained in an infected environment nearly all of these goats would have been expected to develop scrapie. With the exception of the 20 month group, scrapie occurred at an incidence of 25 to 100% in ali other groups and at the expected age. A further observation was that 4 of the progeny of these dairy goats, born and kept apart from any sheep, developed scrapie which suggested that goats were not "dead- end hosts" insofar as scrapie was concerned.
Table 7 recorded the fate of progeny of certain selected scrapied or free Suffolk sheep or dairy goat dams.’
Suffolk ewe G298 was scrapied at 46 months of age. She had twin lambs in 1969 and 1 lamb in 1970. All 3 lambs developed scrapie. Suffolk ewe G27a was scrapied at 39 months. Her lamb born in 1966 was scrapied at 53 months; however, her lambs born in 1967 and 1968 remained free--lived to 102 months of age.
Suffolk ewe G25a died at 131] months of age and was nega- tive clinically and histologically. Mice remained negative following intracerebral inoculation of brain, spleen, and lymph nodes from this ewe. This ewe had 9 progeny at risk, of which 4 developed scrapie and 5 did not. There was no dis- cernible pattern to the cases. In two instances, 1 twin was scrapied and 1 remained free.
Goat B259 was scrapied when 43 months old. All of her 6 progeny at risk developed scrapie.
Goat B14a remained free and died at 101 months of age. Of her 11 progeny at risk, 7 were scrapied and 4 were not.
It was observed at the Station that when scrapied dams had several progeny at risk, 1 or more progeny usually developed the disease. However, many such scrapied dams also had progeny which lived, or are living, considerably beyond the age of their dams and beyond the age animals born at the Station manifested the disease.
It was also observed that individual free dams had free progeny in earlier years followed by scrapied progeny when they were older, or had scrapied progeny when young followed by free progeny when older, or scrapie and free progeny dis- persed throughout the dam's breeding life. The same situation occurred in progeny of scrapied dams; however, the pattern was less irregular due to the smaller number of progeny from each scrapied dam and the higher incidence of scrapie in such progeny. Circumstances prevented breeding all ewes ary year and, thus, many had only 1 progeny at risk. Scrapie developed in 100% of the single progeny at risk of 11 scrapied and 15 free dams. The 26 scrapied progeny were equally divided between ewes and rams.
Table 8 reflected the difference in age scrapied of - sheep brought to the Station compared to the age scrapied of those born there. Although the average age of previously exposed sheep (Suffolks) brought to the Station did not differ greatly from the overall average, several animals brought to the Station developed the disease at quite advanced ages. The previously unexposed scrapied animals brought to the Station were also considerably older than animals born there. Progeny of scrapied dams developed the disease at a slightly younger age than did progeny of free dams. The average age was nearly the same for males and females.
DISCUSSION
snip...see full text;
Tuesday, May 31, 2022
89th General Session of the World Assembly of OIE Delegates image for WOAH General Summit 2022 Chronic Wasting Disease CWD TSE Prion Discussions and Concerns
FRIDAY, NOVEMBER 25, 2022
USA National Scrapie Eradication Program (NSEP) 2021 to 2003 A Year by Year Review
WEDNESDAY, MARCH 16, 2022
SHEEP BY-PRODUCTS AND WHAT ABOUT Scrapie TSE PrP and Potential Zoonosis?
IBNC Tauopathy or TSE Prion disease, it appears, no one is sure
Terry S. Singeltary Sr., 03 Jul 2015 at 16:53 GMT
PLOS ONE Journal
IBNC Tauopathy or TSE Prion disease, it appears, no one is sure
Terry S. Singeltary Sr., 03 Jul 2015 at 16:53 GMT
***however in 1 C-type challenged animal, Prion 2015 Poster Abstracts S67 PrPsc was not detected using rapid tests for BSE.
***Subsequent testing resulted in the detection of pathologic lesion in unusual brain location and PrPsc detection by PMCA only.
*** IBNC Tauopathy or TSE Prion disease, it appears, no one is sure ***
WEDNESDAY, DECEMBER 23, 2020
Idiopathic Brainstem Neuronal Chromatolysis IBNC BSE TSE Prion a Review 2020
Monday, November 14, 2022
Prion Diseases in Dromedary Camels (CPD) 2022 Review
Tuesday, April 27, 2021
Working Document on Camel Prion Disease (CPrD) 14/09/2020
SATURDAY, SEPTEMBER 24, 2022
Transmission of CH1641 in cattle
WEDNESDAY, DECEMBER 8, 2021
Importation of Sheep, Goats, and Certain Other Ruminants AGENCY: Animal APHIA, USDA, FINAL RULE [Docket No. APHIS–2009–0095] RIN 0579–AD10
WEDNESDAY, MARCH 24, 2021
USDA Animal and Plant Health Inspection Service 2020 IMPACT REPORT BSE TSE Prion Testing and Surveillance MIA
https://animalhealthreportpriontse.blogspot.com/2021/03/usda-animal-and-plant-health-inspection.html
SUNDAY, MARCH 21, 2021
Investigation Results of Texas Cow That Tested Positive for Bovine Spongiform Encephalopathy (BSE) Aug. 30, 2005 Singeltary's Regiew 2021
FRIDAY, APRIL 1, 2022
USDA TAKES THE C OUT OF COOL, what's up with that?
THURSDAY, AUGUST 20, 2020
Why is USDA "only" BSE TSE Prion testing 25,000 samples a year?
THURSDAY, JANUARY 23, 2020
USDA Consolidates Regulations for NAHLN Laboratory Testing USDA Animal and Plant Health Inspection Service
sent this bulletin at 01/23/2020 02:15 PM EST
WEDNESDAY, APRIL 24, 2019
USDA Announces Atypical Bovine Spongiform Encephalopathy Detection Aug 29, 2018 A Review of Science 2019
Saturday, July 23, 2016
BOVINE SPONGIFORM ENCEPHALOPATHY BSE TSE PRION SURVEILLANCE, TESTING, AND SRM REMOVAL UNITED STATE OF AMERICA UPDATE JULY 2016
Tuesday, July 26, 2016
Atypical Bovine Spongiform Encephalopathy BSE TSE Prion UPDATE JULY 2016
Monday, June 20, 2016
Specified Risk Materials SRMs BSE TSE Prion Program
*** PLEASE SEE THIS URGENT UPDATE ON CWD AND FEED ANIMAL PROTEIN ***
Sunday, March 20, 2016
Docket No. FDA-2003-D-0432 (formerly 03D-0186) Use of Material from Deer and Elk in Animal Feed ***UPDATED MARCH 2016*** Singeltary Submission
SEE MAD COW FEED VIOLATIONS AFER MAD COW FEED VIOLATIONS ;
Tuesday, April 19, 2016
Docket No. FDA-2013-N-0764 for Animal Feed Regulatory Program Standards Singeltary Comment Submission
17 years post mad cow feed ban August 1997
Monday, October 26, 2015
FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEED VIOLATIONS OFFICIAL ACTION INDICATED OIA UPDATE October 2015
Tuesday, December 23, 2014
FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEEDVIOLATIONS OFFICIAL ACTION INDICATED OAI UPDATE DECEMBER 2014 BSE TSE PRION
16 years post mad cow feed ban August 1997 2013
Sunday, December 15, 2013
FDA PART 589 -- SUBSTANCES PROHIBITED FROM USE IN ANIMAL FOOD OR FEED VIOLATIONS OFFICIAL ACTION INDICATED OIA UPDATE DECEMBER 2013 UPDATE
Saturday, August 29, 2009
FOIA REQUEST FEED RECALL 2009 Product may have contained prohibited materials Bulk Whole Barley, Recall # V-256-2009
Friday, September 4, 2009
FOIA REQUEST ON FEED RECALL PRODUCT 429,128 lbs. feed for ruminant animals may have been contaminated with prohibited material Recall # V-258-2009
Thursday, March 19, 2009
MILLIONS AND MILLIONS OF POUNDS OF MAD COW FEED IN COMMERCE USA WITH ONGOING 12 YEARS OF DENIAL NOW, WHY IN THE WORLD DO WE TO TALK ABOUT THIS ANYMORE $$$
MONDAY, FEBRUARY 25, 2019
***> MAD DOGS AND ENGLISHMEN BSE, SCRAPIE, CWD, CJD, TSE PRION A REVIEW 2019
SATURDAY, OCTOBER 8, 2022
Cattle with the EK211 PRNP polymorphism are susceptible to the H-type bovine spongiform encephalopathy agent from either E211K or wild type donors after oronasal inoculation
TUESDAY, NOVEMBER 01, 2022
SEAC Position statement Chronic wasting disease in UK deer January 2005 (updated July 2006) to 2021
TUESDAY, NOVEMBER 1, 2022
SEAC Scientific Steering Committee on TSE Prion
SATURDAY, NOVEMBER 5, 2022
EFSA Network on BSE-TSE Minutes of the 17th meeting Held on 13-14 October 2022
THURSDAY, OCTOBER 22, 2015
Former Ag Secretary Ann Veneman talks women in agriculture and we talk mad cow disease USDA and what really happened
THURSDAY, FEBRUARY 23, 2012
EIGHT FORMER SECRETARIES OF AGRICULTURE SPEAKING AT USDA'S 2012 AGRICULTURE OUTLOOK FORUM INDUCTED INTO USA MAD COW HALL OF SHAME
2020 DECEMBER
WEDNESDAY, DECEMBER 9, 2020
Biden's pick Tom Vilsack Failed Terribly on Mad Cow BSE TSE Prion, why put him back as top Agriculture pick?
SPONGIFORM ENCEPHALOPATHY ADVISORY COMMITTEE
Minutes of the 99th meeting held on 14th December 2007
snip...
ITEM 8 – PUBLIC QUESTION AND ANSWER SESSION 40.
The Chair explained that the purpose of the question and answer session was to give members of the public an opportunity to ask questions related to the work of SEAC. Mr Terry Singeltary (Texas, USA) had submitted a question prior to the meeting, asking: “With the Nor-98 now documented in five different states so far in the USA in 2007, and with the two atypical BSE H-base cases in Texas and Alabama, with both scrapie and chronic 14 © SEAC 2007 wasting disease (CWD) running rampant in the USA, is there any concern from SEAC with the rise of sporadic CJD in the USA from ''unknown phenotype'', and what concerns if any, in relations to blood donations, surgery, optical, and dental treatment, do you have with these unknown atypical phenotypes in both humans and animals in the USA? Does it concern SEAC, or is it of no concern to SEAC? Should it concern USA animal and human health officials?” 41.
A member considered that this question appeared to be primarily related to possible links between animal and human TSEs in the USA.
There is no evidence that sCJD is increasing in the USA and no evidence of any direct link between TSEs and CJD in the USA. Current evidence does not suggest that CWD is a significant risk to human health. There are unpublished data from a case of human TSE in the USA that are suggestive of an apparently novel form of prion disease with distinct molecular characteristics. However, it is unclear whether the case had been further characterised, if it could be linked to animal TSEs or if other similar cases had been found in the USA or elsewhere. In relation to the possible public health implications of atypical scrapie, H-type BSE and CWD, research was being conducted to investigate possible links and surveillance was in place to detect any changes in human TSEs. Although possible links between these diseases and human TSEs are of concern and require research, there is no evidence to suggest immediate public health action is warranted. The possible human health risks from classical scrapie had been discussed earlier in the meeting. Members noted that there are effective channels of discussion and collaboration on research between USA and European groups. Members agreed it is important to keep a watching brief on new developments on TSEs.
snip...
>>>There is no evidence that sCJD is increasing in the USA and no evidence of any direct link between TSEs and CJD in the USA.<<<
TUESDAY, APRIL 05, 2022 2022
American Academy of Neurology Emerging Sciences
Abstract Website
Incidence of Creutzfeldt-Jakob Disease in the United States 1993-2014
https://creutzfeldt-jakob-disease.blogspot.com/2022/04/incidence-of-creutzfeldt-jakob-disease_5.html
TUESDAY, MAY 24, 2022
Texas Creutzfeldt Jakob Disease CJD TSE Prion Update Singeltary FOIA Request Received May 23, 2022
MONDAY, JUNE 14, 2021
Texas Health and Human Services The Department of State Health Services Creutzfeldt Jakob Disease TSE Prion Report 2021?
SUNDAY, MAY 08, 2022
USA National Prion Disease Pathology Surveillance Center Surveillance Update April 11th, 2022
THURSDAY, JUNE 23, 2022
UK Research and analysis Creutzfeldt-Jakob disease (CJD) update (data to end of December 2021) Updated 21 June 2022
TUESDAY, MAY 10, 2022 Concordance of CSF RT-QuIC across the European Creutzfeldt-Jakob Disease surveillance network https://creutzfeldt-jakob-disease.blogspot.com/2022/05/concordance-of-csf-rt-quic-across.htmlTUESDAY, OCTOBER 18, 2022Assessing the Potential Transmissibility of Bovine and Cervid Prions with a Human Prion Protein-based Model ARS RESEARCHDiagnosis and Reporting of Creutzfeldt-Jakob DiseaseSingeltary, Sr et al. JAMA.2001; 285: 733-734. Vol. 285 No. 6, February 14, 2001 JAMA Diagnosis and Reporting of Creutzfeldt-Jakob Disease To the Editor: In their Research Letter, Dr Gibbons and colleagues1 reported that the annual US death rate due to Creutzfeldt-Jakob disease (CJD) has been stable since 1985. These estimates, however, are based only on reported cases, and do not include misdiagnosed or preclinical cases. It seems to me that misdiagnosis alone would drastically change these figures. An unknown number of persons with a diagnosis of Alzheimer disease in fact may have CJD, although only a small number of these patients receive the postmortem examination necessary to make this diagnosis. Furthermore, only a few states have made CJD reportable. Human and animal transmissible spongiform encephalopathies should be reportable nationwide and internationally.. Terry S. Singeltary, Sr Bacliff, Tex 1. Gibbons RV, Holman RC, Belay ED, Schonberger LB. Creutzfeldt-Jakob disease in the United States: 1979-1998. JAMA. 2000;284:2322-2323.http://jama.jamanetwork.com/article.aspx?articleid=1031186Elsevier Editorial System(tm) for The Lancet Infectious DiseasesManuscript DraftManuscript Number:Title: HUMAN and ANIMAL TSE Classifications i.e. mad cow disease and the UKBSEnvCJD only theoryArticle Type: Personal ViewCorresponding Author: Mr. Terry S. Singeltary,Corresponding Author's Institution: naFirst Author: Terry S Singeltary, noneOrder of Authors: Terry S Singeltary, none; Terry S. SingeltaryAbstract: TSEs have been rampant in the USA for decades in many species, and they all have been rendered and fed back to animals for human/animal consumption. I propose that the current diagnostic criteria for human TSEs only enhances and helps the spreading of human TSE from the continued belief of the UKBSEnvCJD only theory in 2007.HUMAN and ANIMAL TSE Classifications i.e. mad cow disease and the UKBSEnvCJD only theory August 2007August 2007HUMAN and ANIMAL TSE Classifications i.e. mad cow disease and the UKBSEnvCJD only theoryTSEs have been rampant in the USA for decades in many species, and they all have been rendered and fed back to animals for human/animal consumption. I propose that the current diagnostic criteria for human TSEs only enhances and helps the spreading of human TSE from the continued belief of the UKBSEnvCJD only theory in 2007. With all the science to date refuting it, to continue to validate this myth, will only spread this TSE agent through a multitude of potential routes and sources i.e. consumption, surgical, blood, medical, cosmetics etc. I propose as with Aguzzi, Asante, Collinge, Caughey, Deslys, Dormont, Gibbs, Ironside, Manuelidis, Marsh, et al and many more, that the world of TSE Transmissible Spongiform Encephalopathy is far from an exact science, but there is enough proven science to date that this myth should be put to rest once and for all, and that we move forward with a new classification for human and animal TSE that would properly identify the infected species, the source species, and then the route.This would further have to be broken down to strain of species and then the route of transmission would further have to be broken down. Accumulation and Transmission are key to the threshold from sub-clinical to clinical disease, and key to all this, is to stop the amplification and transmission of this agent, the spreading of, no matter what strain. In my opinion, to continue with this myth that the U.K. strain of BSE (one strain TSE in cows), and the nv/v CJD (one strain TSE humans) and that all the rest of human TSE are just one single strain i.e. sporadic CJD (when to date there are 6 different phenotypes of sCJD, and growing per Gambetti et al), and that no other animal TSE transmits to humans, to continue with this masquerade will only continue to spread, expose, and kill, who knows how many more in the years and decades to come. ONE was enough for me, My Mom, hvCJD i.e. Heidenhain Variant CJD, DOD 12/14/97 confirmed, which is nothing more than another mans name added to CJD, like CJD itself, Jakob and Creutzfeldt, or Gerstmann-Straussler-Scheinker syndrome, just another CJD or human TSE, named after another human.WE are only kidding ourselves with the current diagnostic criteria for human and animal TSE, especially differentiating between the nvCJD vs the sporadic CJD strains and then the GSS strains and also the FFI fatal familial insomnia strains or the ones that mimics one or the other of those TSE? Tissue infectivity and strain typing of the many variantsManuscriptof the human and animal TSEs are paramount in all variants of all TSE. There must be a proper classification that will differentiate between all these human TSE in order to do this. With the CDI and other more sensitive testing coming about, I only hope that my proposal will some day be taken seriously. ...Terry S. Singeltary Sr. P.O. Box Bacliff, Texas USA 77518 flounder9@verizon.net
DFA 18
COSMETICS
DFA 17
Medicines and medical devices
DFA 16
MID 1995 TO THE FINAL DAYS farmers with BSE, teenagers with cjd,
DFA 15
Monitoring and Enforcement of the SBO Regulations
BSE Inquiry
DFA 15 Monitoring and Enforcement of the SBO Specified Bovine Offal Regulations
DFA 14
Consideration of the Risk from Mechanically Recovered Meat (MRM) in 1989-1990
TUESDAY, AUGUST 1, 2017
BSE INQUIRY DFA 17 Medicines and medical devices
CJD9/10022
October 1994
Mr R.N. Elmhirst Chairman British Deer Farmers Association Holly Lodge Spencers Lane BerksWell Coventry CV7 7BZ
Dear Mr Elmhirst,
CREUTZFELDT-JAKOB DISEASE (CJD) SURVEILLANCE UNIT REPORT
Thank you for your recent letter concerning the publication of the third annual report from the CJD Surveillance Unit. I am sorry that you are dissatisfied with the way in which this report was published.
snip...
I believe that a further statement about the report, or indeed statistical links between CJD and consumption of venison, would increase, and quite possibly give damaging credence, to the whole issue. From the low key media reports of which I am aware it seems unlikely that venison consumption will suffer adversely, if at all.
http://web.archive.org/web/20030511010117/http://www.bseinquiry.gov.uk/files/yb/1994/10/00003001.pdf
5.195 Among occupational groups exposed to BSE, farmers remain unusual in having such an excess over the incidence of CJD for the population as a whole. No cases of CJD have been reported amount veterinarians exposed to BSE. Four people in the meat industry (butchers, abattoirs, rendering plants, etc) have been reported to have vCJD.386 The present evidence has been accepted by some as reassuring in that such occupations may not pose as serious a risk as might have been expected.
This was not simply another farmer but the third farmer......
suspect case of CJD in a farmer who has had a case of BSE in his beef suckler herd.
http://web.archive.org/web/20030331213802/http://www.bseinquiry.gov.uk/files/yb/1995/10/23006001.pdf
cover-up of 4th farm worker ???
http://web.archive.org/web/20030516083454/http://www.bseinquiry.gov.uk/files/yb/1995/10/23006001.pdf
http://web.archive.org/web/20030330175323/http://www.bseinquiry.gov.uk/files/yb/1995/10/20006001.pdf
CONFIRMATION OF CJD IN FOURTH FARMER
now story changes from;
SEAC concluded that, if the fourth case were confirmed, it would be worrying, especially as all four farmers with CJD would have had BSE cases on their farms.
to;
This is not unexpected...
was another farmer expected?
http://web.archive.org/web/20030728074919/http://www.bseinquiry.gov.uk/files/yb/1995/11/13010001.pdf
4th farmer, and 1st teenager
2. snip...
Over a 5 year period, which is the time period on which the advice from Professor Smith and Dr. Gore was based, and assuming a population of 120,000 dairy farm workers, and an annual incidence of 1 per million cases of CJD in the general population, a DAIRY FARM WORKER IS 5 TIMES MORE LIKELY THAN an individual in the general population to develop CJD. Using the actual current annual incidence of CJD in the UK of 0.7 per million, this figure becomes 7.5 TIMES.
3. You will recall that the advice provided by Professor Smith in 1993 and by Dr. Gore this month used the sub-population of dairy farm workers who had had a case of BSE on their farms - 63,000, which is approximately half the number of dairy farm workers - as a denominator. If the above sums are repeated using this denominator population, taking an annual incidence in the general population of 1 per million the observed rate in this sub-population is 10 TIMES, and taking an annual incidence of 0.7 per million, IT IS 15 TIMES (THE ''WORST CASE'' SCENARIO) than that in the general population...
http://web.archive.org/web/20030516181226/http://www.bseinquiry.gov.uk/files/yb/1995/01/31004001.pdf
CJD FARMERS WIFE 1989
20 year old died from sCJD in USA in 1980 and a 16 year old in 1981. A 19 year old died from sCJD in France in 1985. There is no evidence of an iatrogenic cause for those cases....
http://web.archive.org/web/20030330212925/http://www.bseinquiry.gov.uk/files/yb/1995/10/04004001.pdf
THE COVER UP OF MAD COW DISEASE IN FARMERS, FARMERS WIVES, AND VICKY RIMMER, THE DAY MAD COW SCIENCE CHANGED $$$
Monday, May 19, 2008
*** SPORADIC CJD IN FARMERS, FARMERS WIVES, FROM FARMS WITH BSE HERD AND ABATTOIRS ***
DOES ANYONE BESIDES ME SEE A PATTERN YET ???
Vickey Rimmer, 16, DID NOT DIE FROM nvCJD, she died from a form of sporadic CJD, whatever the hell that is. and there have been 16 year old die from sporadic CJD in the USA as well.
SIMPLY PUT, the ukbsenvcjd only theory was wrong from day one. the elderly are expendable, pets and kids are not.
Science was dictated by 'big buisness' after the Vickey Rimmer case with the ukbsenvcjd only myth.
and there have been 16 year old die from sporadic CJD in the USA as well.
snip...
I have interviewed Mrs Rimmer at my constituency surgery
IF there is nothing to hide, why is there so much SECRECY? WHY is the Government and other Bodies trying to stop any CHANCE OF PEOPLE CONNECTING THE TWO DISEASES. The B.S.E. problem is obvious, but if the correct measures are taken, surely the problem could be contained, however, as it stands the lack of investigation and interest of the possibility of B.S.E. and C.J.D. being linked is open for speculation and surely someone has to account for peoples lives! WHY is so much trouble being taken to convice people that B.S.E. and C.J.D. are not linked? Guilty Conscience perhaps ? - or cover up?
HOUSE OF COMMONS
FROM BARRY JONES, M.P.
22 FEBRUARY 1994
http://web.archive.org/web/20040526010710/http://www.bseinquiry.gov.uk/files/yb/1994/02/22009001.pdf
Alleged Case of Creutzfeld Jakob Disease: Victoria Rimmer.
(now story changes that biopsy shows she does not have CJD...tss)
http://web.archive.org/web/20030511045541/http://www.bseinquiry.gov.uk/files/yb/1994/06/06004001.pdf
now story changes to ;
Advice
7. The Parliamentary Secretary is invited to note the recent statements made on __________ and the present position which remains that CJD cannot be confirmed, in this case at this stage.
http://web.archive.org/web/20030510165315/http://www.bseinquiry.gov.uk/files/yb/1994/06/08004001.pdf
3. The Medical Director at ___________________ Hospital advised the Department on 6 June that the results of ___________________ brain biopsy had been received and that it showed NO EVIDENCE OF CJD. ______________ Hospital subsequently issued a statement to the press to this effect and this was publicised widely in the press (doc 1). News coverage which followed suggested that the statement made by ________________ Hospital had been misleading (doc 2). Enquires have been made of the Medical Director at _______________ Hospital who has CONFIRMED THAT THE STATEMENT ISSUED BY THE HOSPITAL WAS ISSUED IN ERROR. The facts are that two pathology reports on the same piece of brain tissue were recieved. The first report indicated that CJD was unlikely, The second report indicated that CJD was possible, PERHAPS EVEN LIKELY, but that no definitive diagnosis could be made before a post mortem was undertaken.
http://web.archive.org/web/20030511023028/http://www.bseinquiry.gov.uk/files/yb/1994/06/08006001.pdf
MAD COW MEAL DESTROYED MY DAUGHTERS LIFE
A TEENAGE GIRL may have caught the human form of MAD COW DISEASE by eating a contaminated burger it was claimed last night.
VICKY RIMMER, 16, has the killer Creutzfeldt-Jakob disease (CJD).
http://web.archive.org/web/20030510205841/http://www.bseinquiry.gov.uk/files/yb/1994/01/25007001.pdf
GIVE ME BACK MY LIFE
THEY BEGGED ME TO HUSH IT UP – GRAN’S AGONY
http://web.archive.org/web/20040521224258/http://www.bseinquiry.gov.uk/files/yb/1994/01/25008001.pdf
HUSH UP! GOVERNMENT TOLD GRAN: ''YOU MUST THINK OF THE ECONOMY''
http://web.archive.org/web/20031025182000/http://www.bseinquiry.gov.uk/files/yb/1994/01/25009001.pdf
WHY IS MY GIRL DYING ? '' IT WAS LIKE SOMEBODY OLD INSIDE A YOUNG PERSON'S BODY
http://web.archive.org/web/20030513071650/http://www.bseinquiry.gov.uk/files/yb/1994/01/25010001.pdf
see Professor Aguzzi on scjd and bse in Switzerland.
Terry S. Singeltary Sr., Bacliff, Texas USA 77518, Galveston Bay...on the bottom...flounder9@verizon.net
TERRY S. SINGELTARY SR.- retired
- Mr.