Wednesday, March 18, 2009

Detection of CWD Prions in Urine and Saliva of Deer by Transgenic Mouse Bioassay

Detection of CWD Prions in Urine and Saliva of Deer by Transgenic Mouse Bioassay Nicholas J. Haley1, Davis M. Seelig1, Mark D. Zabel1, Glenn C. Telling2, Edward A. Hoover1*

1 Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, United States of America, 2 Department of Molecular Biology and Genetics, University of Kentucky, Lexington, Kentucky, United States of America

Abstract Chronic wasting disease (CWD) is a prion disease affecting captive and free-ranging cervids (e.g. deer, elk, and moose). The mechanisms of CWD transmission are poorly understood, though bodily fluids are thought to play an important role. Here we report the presence of infectious prions in the urine and saliva of deer with chronic wasting disease (CWD). Prion infectivity was detected by bioassay of concentrated, dialyzed urine and saliva in transgenic mice expressing the cervid PrP gene (Tg[CerPrP] mice). In addition, PrPCWD was detected in pooled and concentrated urine by protein misfolding cyclic amplification (PMCA). The concentration of abnormal prion protein in bodily fluids was very low, as indicated by: undetectable PrPCWD levels by traditional assays (western blot, ELISA) and prolonged incubation periods and incomplete TSE attack rates in inoculated Tg(CerPrP) mice (373±3days in 2 of 9 urine-inoculated mice and 342±109 days in 8 of 9 saliva-inoculated mice). These findings help extend our understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections.

Citation: Haley NJ, Seelig DM, Zabel MD, Telling GC, Hoover EA (2009) Detection of CWD Prions in Urine and Saliva of Deer by Transgenic Mouse Bioassay. PLoS ONE 4(3): e4848. doi:10.1371/journal.pone.0004848

Editor: Mark R. Cookson, National Institutes of Health, United States of America

Received: November 8, 2008; Accepted: February 3, 2009; Published: March 18, 2009

Copyright: © 2009 Haley et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: This work was supported by NIH/NCRR Ruth L. Kirschstein Institutional T32 R07072-03 and NIH/NIAID NO1-AI-25491-02 (EAH, GCT). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

* E-mail: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000396/!x-usc:mailto:Edward.Hoover@colostate.edu

SNIP...

Discussion The salient feature of chronic wasting disease is its facile transmission among its host species. Until recently, little was known regarding the mechanisms of this efficient transmissibility, however, we have previously demonstrated infectious prions in the saliva and blood of infected deer [6]. By using intracerebral inoculation of concentrated urine in cervid PrP transgenic mice, we report the presence of infectious prions in urine from CWD-infected cervids, and confirm the phenomenon of prionsialia in these animals. The identification of CWD prions in bodily fluids described in the current report could portend infectivity in secretions and excretions in other prion diseases.

In contrast to the data presented here, oral inoculation of urine in cervid bioassays was unable to identify infectious prions in the urine of CWD+ deer [6]. This result could have been due to necessarily limited observation period possible in those studies (18 months), or variations in source and recipient genotype [14], [15], route of inoculation [16], or the sensitivity of traditional PrPCWD detection assays [17], [18]. The mule deer providing inoculum pools in prior studies were of an unreported genotype; the majority of the recipient deer were homozygous for glycine at residue 96, although a single animal was heterozygous; sharing both G96 and S96 alleles [6]. Likewise, the inocula used in the present study were pooled from sources heterogeneous at codon 96 of the cervid prion gene. Transgenic mice used in bioassay studies, on the other hand, were uniformly homogenous for a glycine residue at this position [9], a polymorphism which is reported to be overrepresented in CWD-infected deer [19]. As a result, it is possible that the genotypic background of either source or subject animals may have been a factor in susceptibility, though we are at present unable to draw any concrete conclusions regarding this relationship. While mouse genotype may have played a role in the outcome, it is also probable that cervid PrP transgenic mouse bioassay simply represents a more sensitive detection system for prions in excreta. Intracranial inoculation, reportedly a more sensitive route of prion exposure [16], [20], is more easily performed in mouse bioassay, a model which also permits extended incubation periods and inclusion of a greater number of test animals.

While our findings point to urine as an additional vehicle for CWD transmission, only 2 of 9 inoculated tg1536 mice were confirmed WB/IHC-positive for prion infection, with a third PrPCWD+ animal later identified by PMCA. This contrasts with 8 of 9 positive mice receiving saliva and infers a much lower concentration of prion infectivity in urine. The wide range of survival times in inoculated mice suggests relatively low levels of infectious prions and/or uneven distribution of infectious PrP moieties in the inocula [21]. Differing [CerPrP] zygosity in tg1536 mice (homozygous vs. hemizygous) may also have played a role in this variation.

Using sPMCA, PrPCWD was repeatedly identified in test urine and spiked urine and saliva used as positive control, but was not detected in test saliva after three rounds of amplification. The reasons for our inability to identify PrPCWD in saliva – given the definitive bioassay findings – remain unknown, and we propose the presence of as-yet unidentified inhibitors such as mucin or salivary proteases which are thought to negatively affect other in vitro assays [22], [23].

The finding of PrPCWD in urine and saliva calls for the identification of the pathological processes and cellular associations of the prion protein involved in shedding. Previous studies have related renal pathology to prionuria [24], [25], a finding which corresponds to our identification of mild to moderate nephritis in those deer providing samples for the current study. It is plausible that renal pathology contributed to prionuria in each of these animals; as samples were pooled, however, we cannot identify specific animals in which it may have been occurring, nor can we accurately estimate the relative level of prionuria occurring in each donor as ultrastructural studies were not performed [26]. While we have not yet identified pathologic prions in renal source tissues [Unpublished data], protease-resistant PrPCWD has been identified by immunostaining in renal tissue of prion-infected deer [27], sheep [28], hamsters and most intriguingly humans [29], foreshadowing the potential for prionuria in other transmissible spongiform encephalopathies. We continue to examine tissues from CWD+ deer in an effort to determine the pathogenesis and kinetics of CWD prion excretion and shedding.

Evidence for excretion and shedding of infectious prions is also accumulating in the scrapie system. PrPC-converting activity has been identified by sPMCA in the urine of scrapie-infected sheep, hamsters and mice [21], [30], [31], [32]. Prion infectivity has also been demonstrated in the feces of hamsters orally infected with scrapie [33]. Other studies point to infectious prions in the milk of scrapie-infected ewes [34], [35]. As noted above, it remains unknown whether other prion diseases (e.g. Kuru, BSE, CJD, TME) may be transmitted by bodily fluids or excreta other than blood. Additional studies examining feces, milk, and other body fluids are therefore necessary in CWD and other prion diseases, studies currently underway in our laboratory.

As CWD transmission may model communicability of other TSE's, the transmissible nature of prion diseases may serve as a model for other protein-misfolding diseases. For example, feces, but not urine, from both mice and cheetahs affected with systemic amyloidosis A (SAA) was recently shown to induce SAA in a mouse model, although negative controls were not available in those studies [36]. In light of the prionuria detected in CWD and in models of scrapie, further investigations of infectivity in body fluids in other protein folding diseases may be warranted in the event that prion diseases are not the only infectious proteinopathies.

In summary, we confirm prionsialia in CWD-affected deer by bioassay in cervidized mice and demonstrate for the first time infectious prions in the urine of these cervids by both bioassay and sPMCA. We are currently evaluating urine and saliva from individual animals in hopes of identifying predisposing factors, such as genotypic background and underlying pathology, which may contribute to prionuria and prionsialia. Concurrently, we have begun to explore the tissue origins and protease sensitivity of the infectious prions as well as the onset and duration of shedding in these bodily fluids.

Acknowledgments

snip...full text ;



http://www.plosone.org/article/fetchObjectAttachment.action;jsessionid=EC743DAAFE1B5688EFD3D7B2A31D4647?uri=info%3Adoi%2F10.1371%2Fjournal.pone.0004848&representation=PDF



http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0004848



http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0004848#s4



Monday, February 09, 2009

Exotic Meats USA Announces Urgent Statewide Recall of Elk Tenderloin Because It May Contain Meat Derived From An Elk Confirmed To Have CWD



http://chronic-wasting-disease.blogspot.com/2009/02/exotic-meats-usa-announces-urgent.html



Wednesday, March 18, 2009 Noah's Ark Holding, LLC, Dawson, MN RECALL Elk products contain meat derived from an elk confirmed to have CWD NV, CA, TX, CO, NY, UT, FL, OK RECALLS AND FIELD CORRECTIONS: FOODS CLASS II



http://chronic-wasting-disease.blogspot.com/2009/03/noahs-ark-holding-llc-dawson-mn-recall.html



Thursday, December 25, 2008

Elk meat recalled due to CWD Boulder County Health Department and Colorado Department of Public Health and Environment



http://chronic-wasting-disease.blogspot.com/2008/12/elk-meat-recalled-due-to-cwd-boulder.html



Saturday, January 24, 2009

Research Project: Detection of TSE Agents in Livestock, Wildlife, Agricultural Products, and the Environment Location: 2008 Annual Report



http://bse-atypical.blogspot.com/2009/01/research-project-detection-of-tse.html





Subject: CWD/POTENTIAL SOURCE/URINE/HUNTERS ? (Mrs. Doe Pee Doe in Estrus)Date: Sun, 14 Jul 2002 08:42:51 -0700
From: "Terry S. Singeltary Sr."
Reply-To: Bovine Spongiform Encephalopathy
To: BSE-L@uni-karlsruhe.de

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

1: Hum Reprod 2002 Jul;17(7):1676-80
Bye-bye urinary gonadotrophins?: Is there a risk of prion diseaseafter the administration of urinary-derived gonadotrophins?

Balen A.

Department of Reproductive Medicine, The General Infirmary, LeedsLS2 9NS, UK. E-mail: adam.balen@leedsth.nhs.uk

Concern has been raised recently about the possibility of prionproteins appearing in the urine of animals and, possibly, humansaffected by prion disease [scrapie, bovine spongiform encephalopathy(BSE) and Creutzfeldt Jakob disease (CJD)]. A debate has started inwhich the suggestion has been made that the purification of human urinefor the provision of gonadotrophins should be discontinued. Thealternative would be to use recombinantly-derived gonadotrophinpreparations. The recombinant products, however, rely upon bovine serumduring the cell culture process and could potentially also be exposed toabnormal prion proteins. It is reassuring that the different types ofgonadotrophin preparations that are currently available are producedwith either urine or bovine serum that is sourced from countries that atthe present time appear to be free of BSE and new variant CJD. We cantherefore be reassured that the gonadotrophins that we usetherapeutically appear to be equally safe.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12093821&dopt=Abstract



Greetings List,


besides the _animal protein_ in deer/elk feed, and the CWDinfected road-kill that goes to render to be manufacturedinto feed, not to mention the Scrapie infected sheep of thepast, and Lord only knows about the cattle, but what aboutthe 100% deer urine they use to atract deer ?
just one example of many below;


CWD/POTENTIAL SOURCE/URINE/HUNTERS ?


Mrs. Doe Pee Doe in Estrus


Model FDE1 Mrs. Doe Pee's Doe in Estrus is made from Estrus urinecollected at the peak of the rut, blended with Fresh Doe Urine for anextremely effective buck enticer. Use pre-rut before the does come intoheat. Use during full rut when bucks are most active. Use duringpost-rut when bucks are still actively looking for does. 1 oz.
http://www.gamecalls.net/huntingproducts/deerlures.html


ELK SCENT/SPRAY BOTTLE
*
Works anytime of the year*
100 % Cow Elk-in-Heat urine (2oz.)*
Economical - mix with water in spray mist bottle*
Use wind to your advantage
Product Code WP-ESB $9.95
http://www.elkinc.com/Scent.asp


prions in urine?
[PDF] A URINE TEST FOR THE IN-VIVO DIAGNOSIS OF PRION DISEASES


http://www.sigov.si/vurs/PDF/diagnoastika-bse-urin.pdf



TSS



########### http://mailhost.rz.uni-karlsruhe.de/warc/bse-l.html ############


http://www.michiganwalleye.com/forum/printthread.php?t=23313


http://www.wapiti.net/discussion/showPost.cfm?post=10918



now, what about those 'deer scents' of 100% urine', and the prion that is found in urine, why not just pass the prion with the urine to other deer... Mrs. Doe Pee Doe in Estrus Model FDE1 Mrs. Doe Pee's Doe in Estrus is made from Estrus urine collected at the peak of the rut, blended with Fresh Doe Urine for an extremely effective buck enticer. Use pre-rut before the does come into heat. Use during full rut when bucks are most active. Use during post-rut when bucks are still actively looking for does. 1 oz. http://www.gamecalls.net/huntingproducts/deerlures.html ELK SCENT/SPRAY BOTTLE * Works anytime of the year * 100 % Cow Elk-in-Heat urine (2oz.) * Economical - mix with water in spray mist bottle * Use wind to your advantage Product Code WP-ESB $9.95 http://www.elkinc.com/Scent.asp prions in urine? [PDF] A URINE TEST FOR THE IN-VIVO DIAGNOSIS OF PRION DISEASES


http://www.sigov.si/vurs/PDF/diagnoastika-bse-urin.pdf



http://www.fda.gov/OHRMS/Dockets/dailys/03/Jan03/012403/8004be07.html



Subject: DOCKET-- 03D-0186 -- FDA Issues Draft Guidance on Use of Material From Deer and Elk in Animal Feed; AvailabilityDate: Fri, 16 May 2003 11:47:37 -0500From: "Terry S. Singeltary Sr." To: fdadockets@oc.fda.gov


http://madcowfeed.blogspot.com/2008/07/docket-03d-0186-fda-issues-draft.html


http://www.buckmasters.com/bm/Community/Forums/tabid/60/forumid/14/postid/10141/view/topic/Default.aspx



From: TSSSubject: DEER SCENTS BANNED DUE TO CWD TRANSMISSIONDate: April 25, 2007 at 7:18 am PST
Last updated at 4:42 PM on 24/04/07
Deer scents banned Wildlife Act amended to avoid chronic wasting disease

BY BETH JOHNSTON The Daily News

Nova Scotian hunters will have to leave their deer pee at home.
In an effort to stop the contagious, lethal Chronic Wasting Disease from hitting Nova Scotian deer and elk, the Department of Natural Resources is banning the use of deer scents which contain deer bodily fluid.

The disease has been diagnosed in commercial game farms in several states and provinces where the products originate. There are no regulations on the imported scents, which hunters can purchase at WalMart and Canadian Tire.

Hunters often soak cotton balls in the urine from a doe in heat to attract bucks.

Chronic wasting disease – a transmissible neurological disease of deer and elk – is a very serious problem in Western Canada and parts of the United States, said Natural Resources wildlife director Barry Sabean.

“We don’t have it and we don’t want it,” he said.

(For full story, see Wednesday's edition of The Daily News)

http://www.hfxnews.ca/index.cfm?sid=24902&sc=89


BETTER LATE THAN NEVER......TSS


From: TSS (216-119-163-192.ipset45.wt.net)
Subject: SEWING THE SEEDS OF CWD MAD DEER/ELK THROUGH FEEDING, ESPECIALLY FROM ANIMAL PROTEIN !!!
Date: September 12, 2002 at 9:52 am PST
CWD AND STUPID SAFETY TIP & COMMENTS TEXAS & SEWING THE SEEDS OF CWD THROUGH ANIMAL PROTEIN?

Houston Chronicle

TDH

CWD is probably not a zoonotic disease. In other words, there isno evidence that CWD can be passed from infected animal to humans

AND

* Always thoroughly cook meat

http://www.tdh.state.tx.us/zoonosis/diseases/CWD.pdf
http://www.tdh.state.tx.us/zoonosis/



with that said, there is no evidence that it cannot, but my opinion, there is more evidence it can, that it cannot.

AND if you plan on cooking the TSE agents out of the meatas implied above, you had better ash it to 1000 degrees celsius.

New studies on the heat resistance of hamster-adapted scrapie agent: Threshold survival after ashing at 600°C suggests an inorganic template of replication

Paul Brown*, [dagger ] , Edward H. Rau [Dagger ] , Bruce K. Johnson*, Alfred E. Bacote*, Clarence J. Gibbs Jr.*, and D. Carleton Gajdusek§
* Laboratory of Central Nervous System Studies, National Institute of Neurological Disorders and Stroke, and [Dagger ] Environmental Protection Branch, Division of Safety, Office of Research Services, National Institutes of Health, Bethesda, MD 20892; and § Institut Alfred Fessard, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France
Contributed by D. Carleton Gajdusek, December 22, 1999

One-gram samples from a pool of crude brain tissue from hamsters infected with the 263K strain of hamster-adapted scrapie agent were placed in covered quartz-glass crucibles and exposed for either 5 or 15 min to dry heat at temperatures ranging from 150°C to 1,000°C. Residual infectivity in the treated samples was assayed by the intracerebral inoculation of dilution series into healthy weanling hamsters, which were observed for 10 months; disease transmissions were verified by Western blot testing for proteinase-resistant protein in brains from clinically positive hamsters. Unheated control tissue contained 9.9 log10LD50/g tissue; after exposure to 150°C, titers equaled or exceeded 6 log10LD50/g, and after exposure to 300°C, titers equaled or exceeded 4 log10LD50/g. Exposure to 600°C completely ashed the brain samples, which, when reconstituted with saline to their original weights, transmitted disease to 5 of 35 inoculated hamsters. No transmissions occurred after exposure to 1,000°C. These results suggest that an inorganic molecular template with a decomposition point near 600°C is capable of nucleating the biological replication of the scrapie agent.

snip...

http://www.pnas.org/cgi/content/full/97/7/3418

But some scientists advocate stricter measures.

Pierluigi Gambetti, director of the National Prion Disease PathologySurveillance Center at Case Western Reserve University in Cleveland, said all deer should be tested for chronic wasting disease before any processing is done.

"There is no way around it," he said. "Nobody should touch that meat unless it has been tested."

snip...

also, what is TEXAS stance on feeding deer and CWD risk?

but before that, lets look at a few things;

Oral transmission and early lymphoid tropism of chronic wasting disease PrPres in mule deer fawns (Odocoileus hemionus )

Christina J. Sigurdson1, Elizabeth S. Williams2, Michael W. Miller3, Terry R. Spraker1,4, Katherine I. O'Rourke5 and Edward A. Hoover1
Department of Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523- 1671, USA1Department of Veterinary Sciences, University of Wyoming, 1174 Snowy Range Road, University of Wyoming, Laramie, WY 82070, USA 2Colorado Division of Wildlife, Wildlife Research Center, 317 West Prospect Road, Fort Collins, CO 80526-2097, USA3Colorado State University Veterinary Diagnostic Laboratory, 300 West Drake Road, Fort Collins, CO 80523-1671, USA4Animal Disease Research Unit, Agricultural Research Service, US Department of Agriculture, 337 Bustad Hall, Washington State University, Pullman, WA 99164-7030, USA5
Author for correspondence: Edward Hoover.Fax +1 970 491 0523. e-mail ehoover@lamar.colostate.edu

Abstract
Top
Abstract

IntroductionMethodsResultsDiscussionReferences

Mule deer fawns (Odocoileus hemionus) were inoculated orally with a brain homogenate prepared from mule deer with naturally occurring chronic wasting disease (CWD), a prion-induced transmissible spongiform encephalopathy. Fawns were necropsied and examined for PrP res, the abnormal prion protein isoform, at 10, 42, 53, 77, 78 and 80 days post-inoculation (p.i.) using an immunohistochemistry assay modified to enhance sensitivity. PrPres was detected in alimentary-tract-associated lymphoid tissues (one or more of the following: retropharyngeal lymph node, tonsil, Peyer's patch and ileocaecal lymph node) as early as 42 days p.i. and in all fawns examined thereafter (53 to 80 days p.i.). No PrPres staining was detected in lymphoid tissue of three control fawns receiving a control brain inoculum, nor was PrPres detectable in neural tissue of any fawn. PrPres-specific staining was markedly enhanced by sequential tissue treatment with formic acid, proteinase K and hydrated autoclaving prior to immunohistochemical staining with monoclonal antibody F89/160.1.5. These results indicate that CWD PrP res can be detected in lymphoid tissues draining the alimentary tract within a few weeks after oral exposure to infectious prions and may reflect the initial pathway of CWD infection in deer. The rapid infection of deer fawns following exposure by the most plausible natural route is consistent with the efficient horizontal transmission of CWD in nature and enables accelerated studies of transmission and pathogenesis in the native species.

snip...

These results indicate that mule deer fawns develop detectable PrP res after oral exposure to an inoculum containing CWD prions. In the earliest post-exposure period, CWD PrPres was traced to the lymphoid tissues draining the oral and intestinal mucosa (i.e. the retropharyngeal lymph nodes, tonsil, ileal Peyer's patches and ileocaecal lymph nodes), which probably received the highest initial exposure to the inoculum. Hadlow et al. (1982) demonstrated scrapie agent in the tonsil, retropharyngeal and mesenteric lymph nodes, ileum and spleen in a 10-month-old naturally infected lamb by mouse bioassay. Eight of nine sheep had infectivity in the retropharyngeal lymph node. He concluded that the tissue distribution suggested primary infection via the gastrointestinal tract. The tissue distribution of PrPres in the early stages of infection in the fawns is strikingly similar to that seen in naturally infected sheep with scrapie. These findings support oral exposure as a natural route of CWD infection in deer and support oral inoculation as a reasonable exposure route for experimental studies of CWD.

snip...

http://vir.sgmjournals.org/cgi/content/full/80/10/2757



now, just what is in that deer feed? _ANIMAL PROTEIN_


Subject: MAD DEER/ELK DISEASE AND POTENTIAL SOURCES
Date: Sat, 25 May 2002 18:41:46 -0700
From: "Terry S. Singeltary Sr."
Reply-To: BSE-LTo: BSE-L
8420-20.5% Antler Developer For Deer and Game in the wild
Guaranteed Analysis Ingredients / Products Feeding Directions


snip...


_animal protein_


http://www.surefed.com/deer.htm


BODE'S GAME FEED SUPPLEMENT #400A RATION FOR DEERNET WEIGHT 50 POUNDS22.6 KG.
snip...


_animal protein_


http://www.bodefeed.com/prod7.htm


Ingredients
Grain Products, Plant Protein Products, Processed Grain By-Products,Forage Products, Roughage Products 15%, Molasses Products, __Animal Protein Products__, Monocalcium Phosphate, Dicalcium Pyosphate, Salt,Calcium Carbonate, Vitamin A Acetate with D-activated Animal Sterol (source of Vitamin D3), Vitamin E Supplement, Vitamin B12 Supplement,Riboflavin Supplement, Niacin Supplement, Calcium Panothenate, CholineChloride, Folic Acid, Menadione Soduim Bisulfite Complex, PyridoxineHydorchloride, Thiamine Mononitrate, d-Biotin, Manganous Oxide, ZincOxide, Ferrous Carbonate, Calcium Iodate, Cobalt Carbonate, DriedSacchoromyces Berevisiae Fermentation Solubles, Cellulose gum,Artificial Flavors added.


http://www.bodefeed.com/prod6.htm

===================================


MORE ANIMAL PROTEIN PRODUCTS FOR DEER
Bode's #1 Game PelletsA RATION FOR DEERF3153
GUARANTEED ANALYSISCrude Protein (Min) 16%Crude Fat (Min) 2.0%Crude Fiber (Max) 19%Calcium (Ca) (Min) 1.25%Calcium (Ca) (Max) 1.75%Phosphorus (P) (Min) 1.0%Salt (Min) .30%Salt (Max) .70%

Ingredients

Grain Products, Plant Protein Products, Processed Grain By-Products,Forage Products, Roughage Products, 15% Molasses Products, __Animal Protein Products__, Monocalcium Phosphate, Dicalcium Phosphate, Salt,Calcium Carbonate, Vitamin A Acetate with D-activated Animal Sterol (source of Vitamin D3) Vitamin E Supplement, Vitamin B12 Supplement,Roboflavin Supplement, Niacin Supplement, Calcium Pantothenate, CholineChloride, Folic Acid, Menadione Sodium Bisulfite Complex, PyridoxineHydrochloride, Thiamine Mononitrate, e - Biotin, Manganous Oxide, ZincOxide, Ferrous Carbonate, Calcium Iodate, Cobalt Carbonate, DriedSaccharyomyces Cerevisiae Fermentation Solubles, Cellulose gum,Artificial Flavors added.

FEEDING DIRECTIONSFeed as Creep Feed with Normal Diet

http://www.bodefeed.com/prod8.htm

INGREDIENTS

Grain Products, Roughage Products (not more than 35%), Processed GrainBy-Products, Plant Protein Products, Forage Products, __Animal Protein Products__, L-Lysine, Calcium Carbonate, Salt, Monocalcium/DicalciumPhosphate, Yeast Culture, Magnesium Oxide, Cobalt Carbonate, BasicCopper Chloride, Manganese Sulfate, Manganous Oxide, Sodium Selenite,Zinc Sulfate, Zinc Oxide, Sodium Selenite, Potassium Iodide,Ethylenediamine Dihydriodide, Vitamin E Supplement, Vitamin ASupplement, Vitamin D3 Supplement, Mineral Oil, Mold Inhibitor, CalciumLignin Sulfonate, Vitamin B12 Supplement, Menadione Sodium BisulfiteComplex, Calcium Pantothenate, Riboflavin, Niacin, Biotin, Folic Acid,Pyridoxine Hydrochloride, Mineral Oil, Chromium Tripicolinate

DIRECTIONS FOR USE

Deer Builder Pellets is designed to be fed to deer under rangeconditions or deer that require higher levels of protein. Feed to deerduring gestation, fawning, lactation, antler growth and pre-rut, allphases which require a higher level of nutrition. Provide adequateamounts of good quality roughage and fresh water at all times.

http://www.profilenutrition.com/Products/Specialty/deer_builder_pellets.html



DEPARTMENT OF HEALTH & HUMAN SERVICESPUBLIC HEALTH SERVICEFOOD AND DRUG ADMINISTRATION
April 9, 2001 WARNING LETTER
01-PHI-12CERTIFIED MAILRETURN RECEIPT REQUESTED
Brian J. Raymond, OwnerSandy Lake Mills26 Mill StreetP.O. Box 117Sandy Lake, PA 16145PHILADELPHIA DISTRICT
Tel: 215-597-4390
Dear Mr. Raymond:

Food and Drug Administration Investigator Gregory E. Beichner conductedan inspection of your animal feed manufacturing operation, located inSandy Lake, Pennsylvania, on March 23,2001, and determined that your firm manufactures animal feeds includingfeeds containing prohibited materials. The inspection found significantdeviations from the requirements set forth inTitle 21, code of Federal Regulations, part 589.2000 - Animal ProteinsProhibited in Ruminant Feed. The regulation is intended to prevent theestablishment and amplification of Bovine Spongiform Encephalopathy(BSE) . Such deviations cause products being manufactured at thisfacility to be misbranded within the meaning of Section 403(f), of theFederal Food, Drug, and CosmeticAct (the Act).

Our investigation found failure to label yourswine feed with the required cautionary statement "Do Not Feed to cattleor other Ruminants" The FDA suggests that the statement bedistinguishedby different type-size or color or other means of highlighting thestatement so that it is easily noticed by a purchaser.

In addition, we note that you are using approximately 140 pounds ofcracked corn to flush your mixer used in the manufacture of animalfeeds containing prohibited material. Thisflushed material is fed to wild game including deer, a ruminant animal.Feed material which may potentially contain prohibited material shouldnot be fed to ruminant animals which may become part of the food chain.

The above is not intended to be an all-inclusive list of deviations fromthe regulations. As a manufacturer of materials intended for animalfeed use, you are responsible for assuring that your overall operationand the products you manufacture and distribute are in compliance withthe law. We have enclosed a copy of FDA's Small Entity Compliance Guideto assist you with complying with the regulation... blah, blah, blah...

http://www.fda.gov/foi/warning_letters/g1115d.pdf


===================================================


now, what about those 'deer scents' of 100% urine',and the prion that is found in urine, why not justpass the prion with the urine to other deer...


Mrs. Doe Pee Doe in EstrusModel FDE1 Mrs. Doe Pee's Doe in Estrus is made from Estrus urinecollected at the peak of the rut, blended with Fresh Doe Urine for anextremely effective buck enticer. Use pre-rut before the does come intoheat. Use during full rut when bucks are most active. Use duringpost-rut when bucks are still actively looking for does. 1 oz.

http://www.gamecalls.net/huntingproducts/deerlures.html



ELK SCENT/SPRAY BOTTLE*Works anytime of the year*100 % Cow Elk-in-Heat urine (2oz.)*Economical - mix with water in spray mist bottle*Use wind to your advantage
Product Code WP-ESB $9.95


http://www.elkinc.com/Scent.asp


prions in urine?


[PDF] A URINE TEST FOR THE IN-VIVO DIAGNOSIS OF PRION DISEASES


http://www.sigov.si/vurs/PDF/diagnoastika-bse-urin.pdf



1st, other states stance on feeding deer and CWD risk?


Although there is no proof how CWD spreads from one deer to the next, common sense tells many people that mouth-to-mouth contact is possibly the culprit,? Stroess said.
The feed pile or feeder presents a perfect opportunity for deer to have mouth, nose or saliva contact with deer carrying DWD.

Just as you and I catch a cold from someone who coughs on us or with whom we have close contact, deer likely get some sicknesses the same way,? he said.
As of July 3, both baiting for the purpose of hunting wildlife and feeding of wildlife became illegal in Wisconsin. This means that backyard deer feeders, feed piles, mineral blocks, salt blocks, protein supplement blocks and all other bait is illegal to use for any deer or other wildlife viewing or hunting purposes.


snip...


http://www.wisinfo.com/heraldtimes/news/archive/local_5812834.shtml


Poulter said the ban on feeding is to keeping deer from congregating and transmitting the disease to one another.

The ban includes food, salt, mineral blocks, and other food products with some exceptions. For example, bird and squirrel feeders close to homes and incidental feeding of wildlife within active livestock operations are exempt from the ban.

http://www.zwire.com/site/news.cfm?newsid=4994835&BRD=606&PAG=461&dept_id=172213&rfi=6


The department is banning feeding of wild deer and other wildlife in areas where wild deer are present. The ban includes food, salt, mineral blocks and other food products, with some exceptions. For example, bird and squirrel feeders close to homes and incidental feeding of wildlife within active livestock operations are exempt from the ban.

The rule also bans the importation of hunter-harvested deer and elk carcasses into Illinois, except for deboned meat, antlers, antlers attached to skull caps, hides, upper canine teeth, and finished taxidermist mounts. Skull caps must be cleaned of all brain and muscle tissue.
Officials from the state said should anyone be caught violating the rule, they would be charged with a petty offense and fined $1,000.

For more information about the rule, visit the department's Web site at

http://dnr.state.il.us/legal/rules-status.htm.
http://www.zwire.com/site/news.cfm?newsid=5091636&BRD=1719&PAG=461&dept_id=25271&rfi=6



NOW, what has the media in TEXAS been saying about this type feeding?

here is something from the Houston Chronicle today;

PLANTING SEEDS FOR CWD, TEXAS STYLE...TSS

Sept. 11, 2002, 7:31PM
It's time to plant seeds for deer season

By SHANNON TOMPKINSCopyright 2002 Houston Chronicle

Texas deer hunters always look for an edge -- something to increase their chances of success or improve the health of deer haunting their lease.

That's why they spend piles of money on equipment such as infrared-sensing cameras to monitor trails and feeders, mineral blocks and protein pellets as supplemental feed and spend restless nights figuring where to put a new blind.

And it's why increasing numbers of deer hunters are investing considerable time, money and sweat equity in creating and husbanding food plots.

"You definitely see a lot of interest in putting in food plots, these days," said Clayton Wolf, coordinator of the Texas Parks and Wildlife Department's white-tailed deer programs. "People are spending a lot of time and effort trying to improve their land or leases to benefit deer."
Proof of that is visible along highways leading from Houston any weekend for the next month or so. Pickups pulling trailers holding tractors, Bush Hog or Agri-Five mowers and disk sets are as nearly as common as those piled with four-wheelers, feeders and box blinds.

The next few weeks -- now through the middle of October -- are the heart of the planting season for deer hunters looking to sow the seeds for cool-weather food plots.

Considering the expense of owning or renting a tractor and implements, buying seed and fertilizer and the physical work involved in putting in food plots, hunters should approach the effort with some planning and knowledge.

Without it, many of those food plots will disappoint their planters, failing to produce the wished-for lush, green carpets and the regular visits by whitetails.

Wolf, who has put in many food plots as a serious East Texas deer hunter and studied the process as part of his profession, has some ideas and input for hunters planning their fall planting.

·Put food plots in the right places:

The idea of food plots is to make the opening attractive to deer. To do that, it has to be a place that offers easy access and security as well as something to eat.

Best places for food plots are adjacent to travel corridors or other thick cover. Corners of fields and other large openings are good choices, as are right-of-ways, fire lanes and old logging roads.
Make certain the area to be planted receives enough sunlight. This is a particular problem in East Texas where many food plots are placed in openings surrounded by heavy forest.

"You need to get a minimum of four to six hours of good sunlight a day on a plot," Wolf said. "The more, the better."

Look for fire lanes, pipelines or other openings that run east-west, Wolf suggested. They'll get a lot more direct sunlight than those running north-south.

·Don't make plots too small:

Many wildlife managers suggest food plot size of at least an acre.

But clearing and planting food plots the size of a football field is impractical for most deer hunters, particularly in East Texas.

Smaller plots will work, but with caveats. Small plots are very susceptible to being "annihilated" by deer before they become established, Wolf said.

"A lot of people think their food plots didn't `make,' when what really happens is the deer hit them so hard early on that they just destroy them," he said.

·Don't plant too early:

Early September typically is hot and dry -- not prime conditions for planting anything.
Also, if hunters mow and disk too early, undesirable plants can take over the plot.

Best bet is to wait until weather turns a bit cooler, usually by late-September, because it slows the growth of warm-weather plants.

"I really like to wait until October to plant my food plots," Wolf said. "You tend to have cooler weather that holds down the weeds and soil moisture usually is better."

·Do soil tests:

East Texas soil typically is hideous acidic and needs help to produce decent stands of forage in food plots.

Soil tests give hunters information about what their soil will produce and what fertilizers or other substances need to be applied for best production for the plants they plan to use.
Soil tests are inexpensive ($10-15) and easily conducted. For hunters in East Texas, the soil laboratory at Stephen F. Austin State University in Nacogdoches is the most accessible Information and forms for soil testing can be found at the lab's Web site,
www.sfasu.edu/ag/soils/.

·Prepare a proper seed bed:

Getting a piece of ground ready to plant involves more than just mowing and disking.

Typically, East Texas soils are so acidic that agriculture consultants suggest applying two tons of lime per acre to get the soil pH near neutral.

That's seldom possible, physically or monetarily, for most hunters. But a good liming, even if at less than 4,000 pounds per acre, is a big plus.

Also, it helps to apply a good general fertilizer -- 13-13-13 is the most common combination.
·What to plant?

Providing seeds for deer food plots has become big business in the past few years. Several "special" seed mixtures are marketed to deer hunters, most of them promising bigger bucks to hunters who use them.

Truth is, the only big bucks produced by the "special" seed mixtures are the ones going into packagers' pockets. The same seeds can be bought in "generic" packaging for far less money.
A mixture of small cereal grains (oats, winter wheat, ryegrass) and one of several varieties of clover is a good choices for cool-weather food plots in East Texas and much of the rest of the state, Wolf said.

Some hunters add turnips, Austrian peas or iron clay peas or other cool-weather plants to their mix.

The cereal grains come on early, providing forage through the early part of deer season. But oats, particularly, shrivel once freezing weather hits.

That's when clover comes on. Clover provides good late-season forage, and really comes into its own in late-winter and early spring.

Clover's spring growth can be very important for deer, particularly bucks, Wolf said. Once bucks drop their antlers and are rebuilding for the coming year, they'll hit the high-protein clover hard, he said.

Good choices for Texas are crimson clover and a new arrowleaf clover developed by Texas A&M. That clover variety, Apache, is more resistant than other types of arrowleaf to wilt and other diseases.

Find out more about Apache at http://overton.tamu.edu/clover/.

Oats and such should be lightly disked when planted.

Clover does best if simply broadcast, then lightly pushed into the soil. Running a four-wheeler over a plot after broadcasting clover works fine, Wolf said.

·Seek professional help:

Technical guidance biologists with TPWD's wildlife division are professionals at helping folks improve their land for wildlife. They can advise hunters or landowners on how best to approach creating food plots for deer and other wildlife on their property.

Contact TPWD's regional wildlife office with requests for assistance; contact information is available on the agency's Web page, www.tpwd.state.tx.us.

·Don't expect miracles from food plots:

Deer prefer native forage over food plots. If native forage is abundant, they'll turn their noses up at oats and such, just as they will ignore corn feeders when acorns and other native mast are available.

Also, if the overall quality of a tract's deer habitat on a tract is poor, no food plot is going to solve that problem.

Deer thrive best in places with rich, natural, biological diversity.

Food plots can be a positive for both deer and deer hunters though they are not a panacea. But, truth is, what's around food plots is much more important than what grows in them.

Shannon Tompkins covers outdoor recreation for the Chronicle. His columns appear Thursdays, Fridays and Sundays.

http://www.chron.com/cs/CDA/story.hts/outdoors/1571427


TSS

Subject: MAD DEER/ELK DISEASE AND POTENTIAL SOURCES
Date: Sat, 25 May 2002 18:41:46 -0700
From: "Terry S. Singeltary Sr."
Reply-To: Bovine Spongiform Encephalopathy
To: BSE-L@uni-karlsruhe.de


now, what about those 'deer scents' of 100% urine',and the prion that is found in urine, why not justpass the prion with the urine to other deer...


Mrs. Doe Pee Doe in EstrusModel FDE1 Mrs. Doe Pee's Doe in Estrus is made from Estrus urinecollected at the peak of the rut, blended with Fresh Doe Urine for anextremely effective buck enticer. Use pre-rut before the does come intoheat. Use during full rut when bucks are most active. Use duringpost-rut when bucks are still actively looking for does. 1 oz.


www.gamecalls.net/hunting...lures.html


ELK SCENT/SPRAY BOTTLE
*
Works anytime of the year*
100 % Cow Elk-in-Heat urine (2oz.)*
Economical - mix with water in spray mist bottle*
Use wind to your advantage
Product Code WP-ESB $9.95
www.elkinc.com/Scent.asp


prions in urine?


[PDF] A URINE TEST FOR THE IN-VIVO DIAGNOSIS OF PRION DISEASES


http://www.sigov.si/vurs/PDF/diagnoastika-bse-urin.pdf
http://p079.ezboard.com/fwolftracksproductionsfrm2.showMessage?topicID=54.topic
http://www.fda.gov/ohrms/dockets/dailys/03/Jan03/012403/8004be07.html



tss



Mrs. Doe Pee Doe in Estrus Model FDE1 Mrs. Doe Pee's Doe in Estrus is made from Estrus urine collected at the peak of the rut, blended with Fresh Doe Urine for an extremely effective buck enticer. Use pre-rut before the does come into heat. Use during full rut when bucks are most active. Use during post-rut when bucks are still actively looking for does. 1 oz.

http://www.gamecalls.net/huntingproducts/deerlures.html


ELK SCENT/SPRAY BOTTLE * Works anytime of the year * 100 % Cow Elk-in-Heat urine (2oz.) * Economical - mix with water in spray mist bottle * Use wind to your advantage Product Code WP-ESB $9.95 http://www.elkinc.com/Scent.asp


prions in urine? [PDF] A URINE TEST FOR THE IN-VIVO DIAGNOSIS OF PRION DISEASES

http://www.sigov.si/vurs/PDF/diagnoastika-bse-urin.pdf


TSS


########### http://mailhost.rz.uni-karlsruhe.de/warc/bse-l.html ############


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http://www.biggamehunt.net/forums/archive/o_t__t_17713__view_previous__index.html



http://www.biggamehunt.net/forums/archive/o_t__t_12826__deer-scents-banned-due-to-cwd-transmission.html



CWD, GAME FARMS, AND BAITING


http://chronic-wasting-disease.blogspot.com/2009/01/cwd-game-farms-baiting-and-politics.html





http://chronic-wasting-disease.blogspot.com/2008/11/commentary-crimes-hurt-essence-of.html





ALSO, NOTE MINERAL LICKS A POSSIBLE SOURCE AND TRANSMISSION MODE FOR CWD ;



http://chronic-wasting-disease.blogspot.com/2009/08/third-international-cwd-symposium-july.html





http://www.cwd-info.org/pdf/3rd_CWD_Symposium_utah.pdf





Thursday, September 10, 2009

Experimental oral transmission of CWD to red deer (Cervus elaphus elaphus): early detection and late stage distribution of protease-resistant protein

http://chronic-wasting-disease.blogspot.com/2009/09/experimental-oral-transmission-of.html


Thursday, September 24, 2009

Validation of Use of Rectoanal Mucosa-Associated Lymphoid Tissue for Immunohistochemical Diagnosis of Chronic Wasting Disease in White-Tailed Deer


http://chronic-wasting-disease.blogspot.com/2009/09/validation-of-use-of-rectoanal-mucosa.html




Sunday, October 04, 2009

CWD NEW MEXICO SPREADING SOUTH TO TEXAS 2009


http://chronic-wasting-disease.blogspot.com/2009/10/cwd-new-mexico-spreading-south-to-texas.html




TSS

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