Back to the Past With New TSE Testing ARS RESEARCH

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Agricultural Research/May-June 2009

Back to the Past With New TSE Testing

Transmissible spongiform encephalopathies (TSEs) are rare—but lethal— neurodegenerative disorders that affect a range of mammals, including humans. Bovine spongiform encephalopathy (BSE)—or “mad cow disease”—is one TSE that has had significant economic and public health impact. The TSEs in the United States are found in sheep, goats, elk, and deer.

TSE epidemiology is complicated by the fact that a definitive diagnosis cannot be made until after an animal has died. ARS chemist Eric Nicholson and veterinary medical officer Robert Kunkle have found a way to facilitate postmortem TSE diagnoses—even when tissue samples are in short supply. Proteins called “prions” are produced in all animals. But the development of abnormal prions is believed to prompt the onset of TSE-related damage to brain tissue. If an animal dies from a TSE infection, both abnormal and healthy prions can be found in its body tissues.

Researchers check tissues for abnormal prions with either Western blotting (WB) or immunohistochemistry (IHC). WB is used for fresh or frozen tissues, and IHC is used to test formalin- fixed tissue that has never been frozen. Sometimes only formalin-fixed tissues are available for testing—a situation the BSE surveillance program faced in 2005, when an entire tissue sample was inadvertently preserved in formalin. In that instance, initial IHC results were not conclusive, but there were no other fresh or frozen tissue samples available for WB testing. Nicholson and Kunkle, who work at the National Animal Disease Center in Ames, Iowa, wanted to help scientists avoid similar situations in the future. They found a way to extract and identify abnormal prions in formalin-fixed tissue by using a combination of mild detergent, a series of freeze-boil cycles, and enzyme digestion. Initial results indicate that the accuracy of this method begins to decline 2 years after the tissue is first preserved, and is completely lost at the end of 6 years. “With this technique, we can easily distinguish between tissues from TSE-positive and TSE-negative animals,” Nicholson says. “And it requires only a minimal adaptation of existing Western blotting procedures.”

Nicholson and Kunkle also devised a way to use WB to test for TSE in tissues that had been fixed in formalin and preserved in paraffin. Their results equaled—and at times even exceeded— the effectiveness of WB analysis for tissues that had only been fixed in formalin.

These combined results add to the tools animal scientists can use to study the development and spread of TSEs. Their findings will facilitate WB testing of tissue samples that were originally archived for microscopy examination and should simplify preservation of samples collected in the field.

WB and IHC analyses can also be conducted on the same preserved sample—a breakthrough that could significantly enhance ongoing TSE research in the field and in the lab.—By Ann Perry, ARS.

This research is part of Animal Health, an ARS national program (#103) described on the World Wide Web at www.nps.

Eric M. Nicholson and Robert A. Kunkle are with the USDAARS National Animal Disease Center, 2300 Dayton Ave., Ames, IA 50010; phone (515) 663-7443 [Nicholson], (515) 663-7190 [Kunkle], fax (515) 663-7458, e-mail [email protected]. gov, [email protected]. ?

Chemist Eric Nicholson (left) and veterinary medical officer Robert Kunkle discuss a Western blot image from a TSE test of formalinfixed tissues.

Agricultural Research/May-June 2009

>>> The TSEs in the United States are found in sheep, goats, elk, and deer. <<<


IS Transmissible Mink Encephalopathy i.e. TME, not a Transmissible Spongiform Encephalopathy ?

DID the mad cow in TEXAS (the h-BSE atypical that WAS FINALLY confirmed), and the h-BSE atypical in Alabama, are these not confirmed cases a Transmissible Spongiform Encephalopathy ?

NOW, one could also argue that this was another cases of mad cow disease in Texas, one that was cover-up ;

FOR IMMEDIATE RELEASE Statement May 4, 2004 Media Inquiries: 301-827-6242 Consumer Inquiries: 888-INFO-FDA

Statement on Texas Cow With Central Nervous System Symptoms 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.

Cattle with central nervous system symptoms are of particular interest because cattle with bovine spongiform encephalopathy or BSE, also known as "mad cow disease," can exhibit such symptoms. In this case, there is no way now to test for BSE. But even if the cow had BSE, FDA's animal feed rule would prohibit the feeding of its rendered protein to other ruminant animals (e.g., cows, goats, sheep, bison).

FDA is sending a letter to the firm summarizing its findings and informing the firm that FDA will not object to use of this material in swine feed only. If it is not used in swine feed, this material will be destroyed. Pigs have been shown not to be susceptible to BSE. If the firm agrees to use the material for swine feed only, FDA will track the material all the way through the supply chain from the processor to the farm to ensure that the feed is properly monitored and used only as feed for pigs.

To protect the U.S. against BSE, FDA works to keep certain mammalian protein out of animal feed for cattle and other ruminant animals. FDA established its animal feed rule in 1997 after the BSE epidemic in the U.K. showed that the disease spreads by feeding infected ruminant protein to cattle.

Under the current regulation, the material from this Texas cow is not allowed in feed for cattle or other ruminant animals. FDA's action specifying that the material go only into swine feed means also that it will not be fed to poultry.

FDA is committed to protecting the U.S. from BSE and collaborates closely with the U.S. Department of Agriculture on all BSE issues. The animal feed rule provides crucial protection against the spread of BSE, but it is only one of several such firewalls. FDA will soon be improving the animal feed rule, to make this strong system even stronger.


I wouldn't doubt that the APHIS/USDA et al even try to persuade the WHO at the next meeting to exempt atypical BSE in the USA as with the Nor-98 atypical scrapie they are trying to get exempt. they are so out of touch with reality on the true extent of mad cow disease in the USA that i think they now believe their own lies. ...TSS

Saturday, May 2, 2009


Tuesday, April 28, 2009 Nor98-like Scrapie in the United States of America ... es-of.html

hmm, were these not Transmissible Spongiform Encephalopathy cases ???


In April of 1985, a mink rancher in Wisconsin reported a debilitating neurologic disease in his herd which we diagnosed as TME by histopathologic findings confirmed by experimental transmission to mink and squirrel monkeys.

The rancher was a ''dead stock'' feeder using mostly (>95%) downer or dead dairy cattle and a few horses. Sheep had never been fed. We believe that these findings may indicate the presence of a previously unrecognized scrapie-like disease in cattle and wish to alert dairy practitioners to this possibility.



Over the next 8-10 weeks, approximately 40% of all the adult mink on the farm died from TME. Since previous incidences of TME were associated with common or shared feeding practices, we obtained a careful history of feed ingredients used over the past 12-18 months. ***The rancher was a “dead stock” feeder using mostly (>95%) downer or dead dairy cattle and a few horses. Sheep had never been fed.***


Saturday, May 2, 2009


Wednesday, February 11, 2009 Atypical BSE North America Update February 2009 Both of the BSE cases ascertained in the US native-born cattle were atypical cases (H-type), which contributed to the initial ambiguity of the diagnosis. 174, 185 In Canada, there have been 2 atypical BSE cases in addition to the 14 cases of the classic UK strain of BSE2: one was the H-type, and the other was of the L-type.198 snip...end source : Enhanced Abstract Journal of the American Veterinary Medical Association January 1, 2009, Vol. 234, No. 1, Pages 59-72 Bovine spongiform encephalopathy Jane L. Harman, DVM, PhD; Christopher J. Silva, PhD ... a.234.1.59

Atypical BSE North America Update February 2009 ... pdate.html

I ask Professor Kong ;

Thursday, December 04, 2008 3:37 PM Subject: RE: re--Chronic Wating Disease (CWD) and Bovine Spongiform Encephalopathies (BSE): Public Health Risk Assessment

''IS the h-BSE more virulent than typical BSE as well, or the same as cBSE, or less virulent than cBSE? just curious.....''

Professor Kong reply ;


''As to the H-BSE, we do not have sufficient data to say one way or another, but we have found that H-BSE can infect humans. I hope we could publish these data once the study is complete. Thanks for your interest.''

Best regards, Qingzhong Kong, PhD Associate Professor Department of Pathology Case Western Reserve University Cleveland, OH 44106 USA


I look forward to further transmission studies, and a true ENHANCED BSE/atypical BSE surveillance program put forth testing all cattle for human and animal consumption for 5 years. a surveillance program that uses the most sensitive TSE testing, and has the personnel that knows how to use them, and can be trusted. I look forward to a stringent mad cow feed ban being put forth, and then strictly enforced. we need a forced, not voluntary feed ban, an enhanced feed ban at that, especially excluding blood. we need some sort of animal traceability. no more excuses about privacy. if somebody is putting out a product that is killing folks and or has the potential to kill you, then everybody needs to know who they are, and where that product came from. same with hospitals, i think medical incidents in all states should be recorded, and made public, when it comes to something like a potential accidental transmission exposure event. so if someone is out there looking at a place to go have surgery done, if you have several hospitals having these type 'accidental exposure events', than you can go some place else. it only makes sense. somewhere along the road, the consumer lost control, and just had to take whatever they were given, and then charged these astronomical prices. some where along the line the consumer just lost interest, especially on a long incubating disease such as mad cow disease i.e. Transmissible Spongiform Encephalopathy. like i said before, there is much more to the mad cow story than bovines and eating a hamburger, we must start focusing on all TSE in all species. ...TSS

Month Number of Tests

Feb 2009 -- 1,891

Jan 2009 -- 4,620 ... ults.shtml


Monday, May 4, 2009

Back to the Past With New TSE Testing Agricultural Research/May-June 2009 ... sting.html

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