Prions in intestines and feces


Human victims of Creutzfeldt Jakob Disease (CJD) are also  shedding infectious prions into public sewers, to end up in
both Class B sewage sludge and Class A sludge compost.   Between 2 and 25% of the 4.5 million cases of Alzheimer’s
Disease and senile dementia victims in the US are actually infected with sporadic CJD.  (Manuelidis, et al, 1989; Boller,
et al, 1989, 1995; Bendixen, 1996; Harrison, 1991; Teixeira, 1995;  Warren, et al, 2005).

Prions have been found in the blood and urine of CJD victims.  (Gabizon, et al, 2001; Reichl, et al 2002) .   Undertakers
and medical facilities routinely discharge CJD infected blood and body fluids into public sewers.  (Yale; UC Davis, CDC)

Prions have been found in feces:

"The Journal of Infectious Diseases 2008;198:81–89 © 2008 by the Infectious
Diseases Society of America. All rights reserved.
0022-1899/2008/19801-0015$15.00 DOI: 10.1086/588193

MAJOR ARTICLE

Transmission and Detection of Prions in Feces

Jiri G. Safar,1,2 Pierre Lessard,1 Gültekin Tamgüney,1,2 Yevgeniy Freyman,1 Camille Deering,1 Frederic Letessier,1
Stephen J. DeArmond,1,3 and Stanley
B. Prusiner1,2,4

Incubation times of 140 days and a rate of prion infection of 80%-100% among exposed animals suggested
transmission by feces, probably via coprophagy.= (eating feces)

" Our findings suggest that horizontal transmission of disease among herbivores may occur through the consumption of
feces or foodstuff tainted with prions from feces of CWD-infected cervids and scrapie-infected sheep.

Our findings suggest that horizontal transmission of disease among herbivores may occur through the consumption of
feces or foodstuff tainted with prions from feces of CWD-infected cervids and scrapie-infected sheep."


Prions can cross the intestinal barrier by riding piggyback on ferritin, a protein normally absorbed by the intestine.  
Because ferritin shares considerable homology across species, these data suggest that PrPSc-associated proteins, in
particular ferritin, may facilitate PrPSc uptake in the intestine from distant species, leading to a carrier state in humans.  
(Singh, et al 2004)

“ . . . enteric infection at early as well as later stages of (CJD) disease, and regardless of the route of agent entry,
implicates potential environmental spread by fecal matter.”  (Radebold, et al 2001)

Radebold K, Chernyak M, Martin D, Manuelidis L. 2001.Blood borne transit of CJD from brain to gut at early stages of
infection. BMC Infect Dis 1:20–25.

Journal of Cellular Biochemistry 9999:1–19 (2006)
PROSPECT A 25 nm Virion Is the Likely Cause of
Transmissible Spongiform Encephalopathies

Laura Manuelidis*   Yale Medical School, New Haven, Connecticut 06510

Page 2 - "

"The observation that infected blood carries the infectious agent to the intestinal tract, a direction opposite to that
commonly assumed, also raises the likelihood of shedding these infectious agents in feces, with further environmental
contamination [Radebold et al., 2001]. This is a common mode of spread for many viruses such as the enteroviruses
and hepatitis B.  "

The absence of detectable prions in material that is clearly infectious goes agains Koch’s first requirement that the
agent must always be present. ""  One Creutzfeldt-Jakob disease
(CJD) agent produces 100,000-fold more infectivity than a second CJD strain, yet these brains
show only 10-fold difference in PrP-res, a 10,000-fold discrepancy [Manuelidis, 1998].
Thus many different animal models show PrPres is a poor marker for infectious titers, and a
lack of PrP-res does not preclude infection. In terms of public health then, it is important
to recognize that asymptomatic but infected individuals without detectable PrP-res may continue to spread infection.


http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=59894
Journal List > BMC Infect Dis > v.1; 2001
BMC Infect Dis. 2001; 1: 20.
Published online 2001 October 25.
Copyright © 2001 Radebold et al; licensee BioMed Central Ltd. Verbatim copying and redistribution of this article are
permitted in any medium for any non-commercial purpose, provided this notice is preserved along with the article's
original URL. For commercial use, contact info@biomedcentral.com

Blood borne transit of CJD from brain to gut at early stages of infection
Klaus Radebold,1 Mark Chernyak,1 Daniel Martin,1 and Laura Manuelidis 1

The widespread infection of gastrointestinal lymphoreticular tissues is important epidemiologically because it suggests
there can be shedding of agent into the gut lumen during all stages of infection. This mechanism may be particularly
important if common gastrointestinal infections and chemical irritations lead to superficial epithelial and submucosal
shedding. Similarly, small amounts of agent might be shed through the urine under some circumstances. These
findings raise the concern that if the agent is repeatedly shed into the environment, it may be extremely difficult to
completely eradicate BSE in endemic areas where thousands of cows have been infected. The presence of any of
these agents (BSE, scrapie, CJD) in excreted materials, including cow manure used as fertilizer could be a source for
continuing human and animal spread.
http://www.medicalnewstoday.com/medicalnews.php?newsid=17807

New study shows how mad cow prions hitch a ride into intestine
15 Dec 2004

They piggyback on iron-storing proteins after surviving digestive juices - A new study from the Department of
Pathology at Case Western Reserve University School of Medicine shows that the infectious version of prion proteins,
the main culprits behind the human form of mad cow disease or variant Creutzfeldt-Jakob Disease (vCJD), are not
destroyed by digestive enzymes found in the stomach. Furthermore, the study finds that the infectious prion proteins,
also known as prions, cross the normally stringent intestinal barrier by riding piggyback on ferritin, a protein normally
absorbed by the intestine and abundantly present in a typical meat dish. The study appears in the Dec. 15 issue of the
Journal of Neuroscience.

Prions are a modified form of normal proteins, the prion proteins, which become infectious and accumulate in the
nervous system causing fatal neurodegenerative disease. Variant CJD results from eating contaminated beef products
from cattle infected with mad cow disease. To date, 155 cases of confirmed and probable vCJD in the world have been
reported, and it is unclear how many others are carrying the infection.

According to the study's senior author Neena Singh, M.D., Ph.D., associate professor of pathology, little is known about
the mechanism by which prions cross the human intestinal barrier, which can be a particularly difficult obstacle to cross.

"The mad cow epidemic is far from over, and the continuous spread of a similar prion disease in the deer and elk
population in the U.S. raises serious public health concerns," said Singh. "It is therefore essential to understand how
this disease is transmitted from one species to another, especially in the case of humans where the infectious prions
survive through stages of cooking and digestion."

Using brain tissues infected with the spontaneously occurring version of CJD which is also caused by prions, the
researchers simulated the human digestive process by subjecting the tissue to sequential treatment with digestive
fluids as found in the human intestinal tract. They then studied how the surviving prions are absorbed by the intestine
using a cell model. The prions were linked with ferritin, a cellular protein that normally binds excess cellular iron to store
it in a soluble, non-toxic form within the cell.

"Since ferritin shares considerable similarity between species, it may facilitate the uptake of prions from distant species
by the human intestine,"said Singh."This important finding provides insight into the cellular mechanisms by which
infectious prions ingested with contaminated food cross the species barrier, and provides the possibility of devising
practical methods for blocking its uptake," she said. "If we can develop a method of blocking the binding of prions to
ferritin, we may be able to prevent animals from getting this disease through feed, and stop the transmission to
humans."

Currently, Singh's group is checking whether prions from distant species such as deer and elk can cross the human
intestinal barrier.

The study was supported by National Institutes of Health grants.

Contact: George Stamatis
George.Stamatis@case.edu
216-368-3635
Case Western Reserve University


http://www.biomedcentral.com/1471-2334/1/20

The widespread infection of gastrointestinal lymphoreticular tissues is important epidemiologically because it suggests
there can be shedding of agent into the gut lumen during all stages of infection. This mechanism may be particularly
important if common gastrointestinal infections and chemical irritations lead to superficial epithelial and submucosal
shedding. Similarly, small amounts of agent might be shed through the urine under some circumstances. These
findings raise the concern that if the agent is repeatedly shed into the environment, it may be extremely difficult to
completely eradicate BSE in endemic areas where thousands of cows have been infected. The presence of any of
these agents (BSE, scrapie, CJD) in excreted materials, including cow manure used as fertilizer could be a source for
continuing human and animal spread.
conclusion
In these studies, we provide evidence that blood can carry the CJD agent toward the gastrointestinal tract in a direction
opposite to that commonly assumed. Peripheral PrP pathology can be detected months before significant agent
replication in the brain. The fact that the agent can spread through blood into excretory organs such as gut implicates
potential shedding of agent into the environment with greater epidemic spread.
*************************************************************************************

http://infectioncontrol.ucsfmedicalcenter.org/Infection_Control_Manual/Sec4-11_Human_Prion_Disease_2005.pdf

Section 4.11    Written 4/98    Revised 4/01, 9/01, 11/01, 3/02, 5/02, 12/02, 4/04
Infection Control Manual Reviewed 10/05

INFECTION CONTROL POLICIES AND PROCEDURES FOR PATIENTS WITH
SUSPECTED OR CONFIRMED HUMAN PRION DISEASE (e.g., Creutzfeldt-Jakob Disease [CJD])
*

UNIVERSITY OF CALIFORNIA, SAN FRANCISCO --
PATIENTS WITH CREUTZFELDT JAKOB DISEASE
‘VIII. DISTRIBUTION OF INFECTIVITY OF PRIONS IN THE HUMAN BODY
Table 1. Distribution of infectivity of prions in the human body’
[PAGE 3 OF 21]
MEDIUM or LOW INFECTIVITY *(Listed  alphabetically)”
TISSUE,
:  adipose tissue adrenal gland   blood*   bone marrow  CSF   dental pulp   feces    gingival tissue   heart   
intestine   kidney   liver    lung

*(HS note:   3 people in UK have already contracted vCJD from blood transfusions )
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15893117&dopt=Citation

Med Hypotheses. 2005;65(1):47-54.        Related Articles,   Links          

Risk analysis of ectoparasites acting as vectors for chronic wasting disease.

Lupi O.
The mode of contamination is not known, although direct contact between infected and non-infected animals via saliva,
urine and feces have been considered. Increasing spread of CWD has raised concerns about the potential
transmission to humans and the conversion of human prion protein by CWD-associated prions has been demonstrated
in laboratory experiments

http://www.ncbi.nlm.nih.gov/pubmed/15893117?dopt=Citation


All: 1         Review: 0        Click to change filter selection through MyNCBI.         


1: Med Hypotheses. 2005;65(1):47-54.
Related Articles,  Links

Risk analysis of ectoparasites acting as vectors for chronic wasting disease.

Lupi O.

Department of Medical Clinics, Microcirculation Research Laboratory, Universidade do Estado do Rio de Janeiro, Rua
Frei Leandro, 16\501, 22.470-210 Rio de Janeiro, RJ, Brazil. omarlupi@globo.com

Prion diseases are rare neurodegenerative diseases of humans and animals with a lethal evolution. Animal prion
infections, such as chronic wasting disease (CWD) and scrapie (sheep) have shown a pattern of horizontal
transmission. CWD is an endemic disease that has been affecting thousands of domestic and wild cervids in US for the
last three decades. The mode of contamination is not known, although direct contact between infected and non-
infected animals via saliva, urine and feces have been considered. Increasing spread of CWD has raised concerns
about the potential transmission to humans and the conversion of human prion protein by CWD-associated prions has
been demonstrated in laboratory experiments. Fly larvae exposed to brain infected material were able to readily
transmit scrapie to hamsters. Prion rods were identified in both larvae and fly pupae. New lines of evidence confirmed
that adult flies are also able to express prion proteins. The most prevalent species of myiasis in cattle, sheep and wild
cervids (Hypoderma spp.) present a very different life cycle from human myiasis, with a long contact with neurologic
structures, such as the spinal canal and epidural fat, that are potentially rich in prion rods. Considering the huge
amount of fly larvae that affects each animal, it is important to discuss the possibility that these ectoparasites could
theoretically act as reservoirs and vectors for CWD and other prion diseases. It is critical to recognize all the possible
factors involved in CWD transmission since ectoparasites could be handled in an easier way than the environmental
persistence of infectious prions.

http://www.bseinquiry.gov.uk/report/volume2/chaptea5.htm

3.125 Among untested mechanisms for lateral transmission are faecal and other contamination of pasture, and the
occurrence of an intermediate host. It is known that TSE infectivity can persist after being buried in the soil for at least
three years. 31 There is evidence that field mice and maggots can carry the TSE agent following feeding on infected
cattle waste, and it is possible that either can act to contaminate the feed or pasture provided for cattle.    (Can flies
transmit infectious prions ? )


**************************************************************************************

Blood Transfusion and Spread of Variant Creutzfeldt-Jakob Disease Klaus Dietz,* Gnter Raddatz,* Jonathan
Wallis,† Norbert Mller,‡ Inga Zerr,ァ Hans-Peter Duerr,*Hans Lefvre,カ Erhard Seifried,# and Johannes Lwer**

The model predicts that an infection like vCJD,which has been introduced into the population by the alimentary route,
could not become endemic by transfusion alone and that only <1% of cases would be avoided by excluding from blood
donation those persons who have received a transfusion. ...

[The alimentary canal includes the mouth, pharynx, esophagus, stomach, small intestine, large intestine, and anus.]


http://www.wildlifeinformation.org/Subdirectories_for_Search2/SampleTechs/O_cwd05understprions.htm
The normal route of transmission for the TSEs is thought to be oral. In natural disease PrPSc  can be detected in
lymphoid tissues associated with the alimentary tract before it is detected in nervous tissues and the CNS.
In CWD the disease is known to be transmitted between animals (lateral transmission) and this is thought probably to
be by the oral route.
How the agent leaves the host:
Since PrPCWD can be found in lymphoid tissues associated with the alimentary tract from relatively early in the course
of infection it is possible that secretions and excretions such as saliva and faeces may act as sources of the infectious
agent for other animals to take in orally.


**********************************************************

http://desmoinesregister.com/apps/pbcs.dll/article?AID=/20060618/Bhttp://desmoinesregister.com/apps/pbcs.dll/article?
AID=/20060618/BUSINESS01/606180340/1030USINESS01/606180340/1030

Waste dispute threatens disease lab's work

Workers and others fear poor disposal practices mean animal illnesses the Ames facility studies could reach Iowa's
waters.
"The room in Building 5 where workers perform necropsies has drains on the floor that eventually lead to the Ames
sewage treatment plant and then into the South Skunk River. Workers flush tissue scraps, blood, urine and other items
down the drain into a heated storage tank and then on to the city sewage plant."

""It is known that CWD is an environmental contaminant from work done at Colorado State University," Morgan wrote. "It
most likely is a fecal contamination that occurs, but that is not entirely proven (a good research project). It is known the
sewage treatment does not kill prions, and it is also known that CWD is coming this direction naturally."
******************************************************



CWD – feces

http://www.cdc.gov/ncidod/EID/vol10no6/04-0010.htm

Research
Environmental Sources of Prion Transmission in Mule Deer
Michael W. Miller,*  Elizabeth S. Williams,† N. Thompson Hobbs,‡ and Lisa L. Wolfe*
*Colorado Division of Wildlife, Fort Collins, Colorado, USA; †University of Wyoming, Laramie, Wyoming, USA; and
‡Colorado State University, Fort Collins, Colorado, USA


“Prions cannot be directly demonstrated in excreta or soil. However, CWD infection–specific protease-resistant prion
protein (PrPCWD) accumulates in gut-associated lymphoid tissues (e.g., tonsils, Peyer patches, and mesenteric lymph
nodes) of infected mule deer (11,17,22), which implicates alimentary shedding of the CWD agent in both feces and
saliva (10,11,17).”

“As a result of such behavior, encounters with contaminated environments will occur more frequently than if deer
movements were random. Feces and carcass remains are routinely encountered on native ranges, thus representing
natural opportunities for exposure. Social behavior of deer, particularly their tendency to concentrate and become
sedentary on their winter range, also may increase the probability of coming into contact with sources of infection in
their environment.”
http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=PubMed&list_uids=16544935&dopt=Citation

Jpn J Vet Res. 2006 Feb;53(3-4):149-57.                 
Alymphoplasia mice are resistant to prion infection via oral route.

Horiuchi M, Furuoka H, Kitamura N, Shinagaw M.

Laboratory of Prion Disease, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku, Sapporo 060-
0818, Japan. horiuchi@vetmed.hokudai.ac.jp

”The major cause of infection in animal prion diseases is thought to be consumption of prion-contaminated stuff. There
is evidence that the enteric nerve system (ENS) and gut-associated lymphoid tissues (GATL) are involved in the
establishment of prion infection through alimentary tract. “

“These results indicate that GALT rather than ENS acts as the primary entry port for prion after oral exposure.” = gut-
associated lymphoid tissues (GATL)


Subject: Resistance to CWD in Transgenic Mice Expressing a Naturally Occurring Allelic Variant of Deer
Prion Protein
Date: April 12, 2007 at 9:17 am PST

Resistance to Chronic Wasting Disease in Transgenic Mice Expressing a Naturally Occurring Allelic Variant of Deer
Prion Protein

Kimberly Meade-White,1 Brent Race,1 Matthew Trifilo,2 Alex Bossers,3 Cynthia Favara,1 Rachel Lacasse,1 Michael
Miller,4 Elizabeth Williams,5 Michael Oldstone,2 Richard Race,1 and Bruce Chesebro1*

Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, NIAID, Hamilton, Montana 59840,1

“Extraneural accumulation of PrPres was also noted in spleen and intestinal tissue of clinically ill G96 mice. These
results demonstrate the importance of deer PrP polymorphisms in susceptibility to CWD infection. Furthermore, this
deer PrP transgenic model is the first to demonstrate extraneural accumulation of PrPres in spleen and intestinal tissue
and thus may prove useful in studies of CWD pathogenesis and transmission by oral or other natural routes of
infection.”


CWD Annual Report – 2005
To date, we have developed a reliable, non-lethal method for detecting
chronic wasting disease in  mule deer that represented a significant breakthrough in improving the  ability to monitor
prevalence of the disease over time and space. We also have demonstrated  that chronic wasting disease can be
transmitted indirectly, and that fecal shedding and  fecal-oral transmission is likely.

“Transmission Although the exact method of transmission of CWD is not known, horizontal and indirect transmission
appear to be the most important routes of spread.99 Epidemiologic observations suggest that in addition to
transmission within species, CWD can be transmitted from elk to mule deer and white-tailed deer, from mule
deer to elk, and from mule deer to white-tailed deer.156”
The role of environmental contamination in maintaining infectivity is not entirely understood; however, controlled
studies showed infectivity remained on pastures
in which CWD-affected deer resided approximately 2 years previously. These studies were conducted in pastures
presumed to be highly contaminated. Thus, extrapolation to field situations should be done with care, but these data
suggest caution in managing pastures or
paddocks that have housed CWD-affected cervids

[how about dairy pastures, hay fields and grazing lands topdressed with sewage sludge which may contain infectious
human and animal prions ? ]



http://www.stanford.edu/group/virus/prion/2005priya/newfindings.html

Could other animal TSEs pose a risk to humans?
Belay et al. 2004. Chronic Wasting Disease and Potential Transmission to Humans. Emerging Infectious Diseases Vol.
10, No. 6, June 2004, pg. 977-984.

Chronic wasting disease (CWD) of deer and elk is endemic in a tri-corner area of Colorado, Wyoming, and Nebraska,
and new foci of CWD have been detected in other parts of the United States. Although detection in some areas may be
related to increased surveillance, introduction of CWD due to translocation or natural migration of animals may account
for some new foci of infection. Increasing spread of CWD has raised concerns about the potential for increasing human
exposure to the CWD agent. The foodborne transmission of bovine spongiform encephalopathy to humans indicates
that the species barrier may not completely protect humans from animal prion diseases. Conversion of human prion
protein by CWD associated prions has been demonstrated in an in vitro cell free experiment, but limited investigations
have not identified strong evidence for CWD transmission to humans. More epidemiologic and laboratory studies are
needed to monitor the possibility of such transmissions.

****************************************************************************





SHEEP    SCRAPIE  INTESTINES

Copyright © 2004 Elsevier B.V. All rights reserved.

Involvement of gut-associated lymphoid tissue of ruminants in the spread of transmissible spongiform encephalopathies

Charles McL. Press Corresponding Author Contact Information
Department of Morphology, Genetics and Aquatic Biology, Norwegian School
of Veterinary Science, P.O. Box 8146 Dep., Oslo, Norway

Received 15 September 2003;  accepted 3 November 2003.  Available online
3 February 2004

Scrapie is a transmissible spongiform encephalopathy (TSE) and its
spread across the intestine of sheep is linked to the biology of
intestinal Peyer's patches (PPs).

***************************************************************

February 22, 2006    Also appeared in print February 27, 2006, p. 9 Chemical & Engineering News   ISSN 0009-2347  
Copyright © 2006 American Chemical Society

ENVIRONMENT     Tracking Prions
Method to detect infectious proteins in soils could help monitor spread of the diseases they cause
Steve Ritter
A method to extract and quantitatively detect prions from soil samples has been devised by a team of scientists at two
National Institute for Agricultural Research (INRA) labs in France (Environ. Sci. Technol. 2006, 40, 1497). The
technique could be "a good starting point" to help identify and map prion-contaminated farmland as well as to monitor
the fate of prions over time, notes lead author Peggy Rigou.

Prions are malformed proteins that are thought to be the infectious agents responsible for transmissible spongiform
encephalopathies (TSEs), such as mad cow disease, scrapie in sheep, and chronic wasting disease in deer. Prions
can persist in soil for years, and some animals are suspected of contracting TSEs by drinking water or grazing on
ground that was exposed to the carcasses of dead animals; by-products from animal processing; or animal manure,
urine, or blood.

http://www.drugresearcher.com/news/ng.asp?n=73490-fda-tse-bse-medical-vcjd

FDA cuts cattle material in latest BSE precautions

by Anna Lewcock   18/01/2007
Prohibited Materials:
The brain, skull, eyes and spinal cords from cattle 30 months and older;
The tonsils and a portion of the small intestines from all cattle regardless of their age or health;
************************************************************

Dr Michael Greger, Organic Consumers Association– April 2004:

“One proposal is to exclude cattles' small intestines from human  consumption in the U.S.  In Europe, though, all of the
intestines are  excluded from human food, from the small intestine down to the  rectum,[1] in part because there is
concern that the colon may also  be infectious.[2] Tell the USDA that they should follow Europe's  example and exclude
all cow and calf rectum, colon, and anus from the American food supply.”

*****************************************************************************

http://www.cnn.com/HEALTH/library/DS/00531.html

Creutzfeldt-Jakob disease
From MayoClinic.com
Special to CNN.com

Choose the right cuts. In the United States and elsewhere, avoid the highest risk parts of cattle, such as the eyes,
brain, spinal cord and intestine.   You may feel more comfortable avoiding hot dogs, hamburger, sausages and
luncheon meats. Keep in mind that natural sausage casings made from sheep intestines also may put you at risk.


http://www.organicconsumers.org/madcow/base21604.cfm

New mad cow strain similar to human CJD

February 16, 2004 United Press International
WASHINGTON, Feb. 16 (UPI) -- Italian researchers said Monday they have discovered a new strain of mad cow
disease that is very similar to a spontaneously occurring form of a deadly human brain disorder called sporadic
Creutzfeldt-Jakob disease.

Consumers have some degree of protection because the USDA requires the brain and spinal cord -- the most
infectious parts of a cow -- to be removed and not allowed into human food. Hansen, however, noted bits of brain and
spinal cord have been found in hamburger meat processed by mechanical devices. Other infectious parts of the
animal, such as bone marrow and segments of the intestine, have also been allowed for human consumption.

http://ceresnet.org/images%5CMisc%5CCFNP_TSE_Proceedings.pdf


"Oral exposure may be important because CWD prions are first found in gastrointestinal tract lymphoid tissues. This
suggests that prions are shed in feces and/or saliva.
Page 11 - "Dr. Joel Pederson from the University of Wisconsin-Madison noted that prions can survive in the soil for
years. This is a critical factor because livestock and wild animals incidentally and accidentally consume soil."
pg. 78: "Early prion accumulation in GI tract lymphoid tissue is additional evidence that oral exposure is important and
that prions may be shed in feces and saliva."

Subject: Infectious Prions in the Saliva and Blood of Deer with Chronic
Wasting Disease
Date: October 5, 2006 at 1:45 pm PST
Infectious Prions in the Saliva and Blood of Deer with Chronic Wasting Disease

Candace K. Mathiason,1 Jenny G. Powers,3 Sallie J. Dahmes,4 David A.
Osborn,5 Karl V. Miller,5 Robert J. Warren,5 Gary L. Mason,1 Sheila A. Hays,1 Jeanette Hayes-Klug,1   Davis M. Seelig,
1 Margaret A. Wild,3 Lisa L. Wolfe,6 Terry R. Spraker,1,2 Michael W. Miller,6 Christina J. Sigurdson,1Glenn C. Telling,
7 Edward A. Hoover1*
The transmission of CWD by a single blood transfusion from two symptomatic and one asymptomatic CWDþ donor is
important in at least three contexts: (i) It reinforces that no tissue from CWD-infected cervids can be considered free of
prion infectivity; (ii) it poses the possibility of hematogenous spread of CWD, such as through insects; and (iii) it
provides a basis for seeking in vitro assays sufficiently sensitive to demonstrate PrPCWD or alternate prion protein
conformers in blood—one of the grails of prion biology and epidemiology.
“Recent studies have shown that PrPres is poorly preserved after incubation with intestinal or fecal content (23, 24).
Further research using cervid and surrogate cervid PrP transgenic mice (25) are indicated to continue to address the
presence of infectious CWD prions in excreta of CWDþ deer and to provide a more  substantial basis for
reconsideration of the assumption that excreta are the chief vehicle for CWDdissemination and transmission. “

BSE, TSE
http://www.stanford.edu/group/virus/prion/2005priya/newfindings.html

Could infectious prions be distributed in a range of body tissues?

Cunningham et al. 2004. Distribution of Bovine Spongiform Encephalopathy in Greater Kudu. Emerging Infectious
Diseases ¥
www.cdc.gov/eid Vol. 10, No. 6, June 2004.

Contrary to findings in cattle with BSE in which the tissue distribution of infectivity is the most limited recorded for any of
the transmissible spongiform encephalopathies (TSE), infectivity in greater kudu (Tragelaphus strepsiceros) with BSE
is distributed in as wide a range of tissues as occurs in any TSE.

BSE agent was also detected in skin, conjunctiva, and salivary gland, tissues in which infectivity has not previously
been reported in any naturally occurring TSE. The distribution of infectivity in greater kudu with BSE suggests possible
routes for transmission of the disease and highlights the need for further research into the distribution of TSE
infectious agents in other host species.


[
BSE-L] Prions spread via the autonomic nervous system from the gut to the central nervous system in
cattle incubating BSE

Prions spread via the autonomic nervous system from the gut to the central
nervous system in cattle incubating bovine spongiform encephalopathy
Christine Hoffmann, Ute Ziegler, Anne Buschmann, Artur Weber, Leila Kupfer,
Anja Oelschlegel, Baerbel Hammerschmidt and Martin H. Groschup


Institute for Novel and Emerging Infectious Diseases,
Friedrich-Loeffler-Institut, Boddenblick 5a, 17493 Greifswald-Insel Riems,
Germany

Correspondence   Martin H. Groschup     martin.groschup@fli.bund.de

“Our results demonstrate two routes involving the autonomic nervous system through which BSE prions spread by
anterograde pathways from the gastrointestinal tract (GIT) to the central nervous system (CNS): (i) via the coeliac and
mesenteric ganglion complex, splanchnic nerves and the lumbal/caudal thoracic spinal cord
(representing the sympathetic GIT innervation); and (ii) via the Nervus
vagus (parasympathetic GIT innervation).”

http://vir.sgmjournals.org/cgi/content/abstract/88/3/1048?
maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=prion&searchid=1&FIRSTINDEX=0&volume=88&issue=3&r
esourcetype=HWCIT
**************************************************************









************************************************************

UK – SEAC – UK
Spongiform Encephalopathy Advisory Committee bases conclusion that sewage sludge is
no risk on false assumptions.   
  
http://www.seac.gov.uk/papers/paper_inf81-8.pdf

The following question was submitted by Mr Clapp for the SEAC 80th meeting Q&A session:
“How can human waste disposal to farmland be justified with the increasing likelihood of human prion
diseases being the true source of BSE? The continuing number of cases in the UK would suggest the
origin of BSE was not offal but another feed contaminant.”
This question was addressed by correspondance as Mr Clapp was not present at the SEAC meeting.
Correspondance  1. Reply by e-mail from SEAC secretariat    10th December, 2003
Dear Mr Clapp    Thank you for your query to SEAC for the public Q&A. As you werenot present at the meeting itself, I
thought it would be helpful to send you a reply in writing.

With regards to your question “How can human waste disposal to farm land be justified with the increasing likelihood of
human prion diseases being the true source of BSE?”

This concerns the theoretical possibility that infective prion agent may be passed from infected cattle and humans into
sewage, which could lead to re-infection of grazing cattle via the spreading of sewage sludge on farmland.
SEAC has not considered issues specifically related to human sewage sludge disposal on land with respect to the
potential presence of CJD infectivity, but from the guidance issued by theAdvisory Committee on Dangerous
Pathogens (ACDP)/SEAC joint working group, to date, no TSE infectivity has been shown in human faeces or urine
(see Part 4 and Annexes A1 and A2 of the guidance).[ the truth:   Prions have been found in the blood and urine of
CJD victims.  (Gabizon, et al, 2001; Reichl, et al 2002) .   ] The ACDP/SEAC guidance can be found using the following
web link
http://www.doh.gov.uk/cjd/tseguidance/ .

SEAC has considered the practice of spreading sewage sludge on and in the context of the risk of specified risk
material (SRM) particles from abattoir waste entering the sewage system and advised that provided the particulate
matter was retained and disposed of as SRM, SEAC was content for abattoirs to discharge their liquid waste to sewers
and for sewage sludge to be disposed of by spreading on land. Any small particulate matter passing through the trap
would be diluted to such an extent to pose a negligible risk.   [to the contrary, see Aiken, Pedersen, et al – soil
increases prion infective risk by factor of 680 . . . ]

Regarding your comment that “the continuing number of cases in the UK would suggest the origin of BSE was not offal
but another feed contaminant”, SEAC reviewed hypotheses with regard to the origin and transmission of BSE at their
most recent meeting on 26 November 03 and concluded that, in the context of (livestock) born after the reinforced feed
ban (BARB) cases, although feed contamination was a plausible explanation, other hypotheses such as maternal or
horizontal transmission or environmental factors could not be excluded as plausible explanations for at least some of
the BARB cases. A summary of the 26 November SEAC meeting can be found on our website
http://www.seac.gov.
uk/summaries/summ_1103.htm
I hope that this information is of help to you, please contact the secretariat if you require further information.
Yours sincerely, Pat Keep Dr Pat Keep   SEAC secretariat
******************************************************************************
The UK Spongiform Encephalopathy Advisory Committee (SEAC) forwarded a new research article  this past week:

"Interactions of prion proteins with soil", Liviana Leita, et al, Soil Biology & Biochemistry, (Elsevier) accepted
November 7, 2005"
Excerpts:
"Results clearly demonstrate that both the no infectious (PrPc) and infectious (PrPsc) forms are absorbed by all soils. .
. . . The strong interaction of PrPsc with soil favors the accumulation of prions in soils, especially if amended with prion-
containing organic fertilizers and/or whenever TSE-affected animal carcasses, placenta, and excreta in general are
buried or laid at the soil surface."

“If the adsorbed prions also retain their pathological activity (Brown and Gajdusek, 1991; Leita et al., unpublished
results), they could represent a hazardous environmental source of infectivity, and provide a further explanation for the
horizontal transmissibility of TSE forms, such as scrapie and CWD."

*****************************************************

http://www.cdpqinc.qc.ca/Champs_dactivite/04Sante/references/BIOSECURITY1.pdf

"This is because infection causing agents (prions, viruses, bacteria and parasites) can be transmitted in a great variety
of ways, some of them extremely insidious. Infected animals excrete different microbes via different routes. As a rule,
microbes that live in an animal's respiratory tract are airborne, while those that live in the digestive tract are passed in
manure. Some microbes, however, may also be found in semen, saliva, nasal discharge, urine, milk, vaginal discharge,
meat and so on, where they may survive for days or even weeks."

ALIMENTARY SHEDDING

http://www.plos.org/press/plpa-02-04-pedersen.pdf   (last page:  Funding:  This work was supported by USEPA grant . .
. and DOD grant . . . "

Because their work is funded by EPA and DOD, the authors are loathe to use the words "feces" "excreta" or "manure"
because connecting the dots may lead to questions about land application of  sludge/biosolids . . . . . so they dealt with
the issue as follows:

"Soil has been hypothesized to act as a reservoir of infectivity because PrPsc likely enters soil environment through
urinary or alimentary shedding and decomposition of infected animals."

The CDC, et al, are less circumspect and spell it out - alimentary shedding = feces, saliva, urine . . .


CDC - Environmental Sources of Prion Transmission in Mule Deer



CDC - Environmental Sources of Prion Transmission in Mule Deer
Whether transmission of the chronic wasting disease (CWD) prion among cervids ... which implicates alimentary
shedding of the CWD agent in both feces and ...
www.cdc.gov/ncidod/eid/vol10no6/04-0010.htm - 54k - Cached - Similar pages

[

[PDF
] Chronic Wasting Disease in Deer and Elk: Scientific Facts and Findings
File Format: PDF/Adobe Acrobat
Chronic wasting disease (CWD) is a prion disease of cervids such as deer and elk in North ... agent may be shed
through the alimentary tract (feces and ...
www.cwd-info.org/pdf/ CWD%20Review%20Article%20-%20Salman%202003.pdf - Similar pages

Mad deer
It was prompted by concerns that chronic wasting disease (CWD), ... their saliva or urine as the infectious agent is
shed through the alimentary tract. ...
www.pubmedcentral.nih.gov/ articlerender.fcgi?artid=1308310 - Similar pages


Mad deer
'Chronic wasting disease is the most mysterious of all prion diseases' ... saliva or urine as the infectious agent is shed
through the alimentary tract. ...
www.nature.com/embor/journal/v3/n12/full/embor006.html - Similar pages
*******************************************************************************

The Pedersen, et al, study basically confirms what earlier researchers  found -- that healthy animals have contracted
CWD from soil exposure after they are placed in pens previously inhabitated by CWD infected animals.   What is really
worrisome is the following:

"Taken together, these data support the motion that PrPsc-contaminated soil may allow intraspecies TSE
transmission and enhance the likelihood of spread to other species."

Dr. Pedersen, Dr. Trina McMahon, et al, also have a $100,000 grant from EPA to study prions in sludge biosolids . . . .
so far, the EPA  has only acknowledged the potential for infectious animal prions in sludge (BSE and CWD) . . . the
EPA and  waste industry have assiduously avoided acknowledging the potential for infectious human prions in  Class B
AND particulary in the "pathogen-free"  Class A sludge biosolids . . .


MISCELLANEOUS

feces EXCRETA scrapie ferritin summary intestines animals dogs cats pigs sheep no species barrier ? joel pederson -
feces FLIES - ECTOPARASITES CWD CONVERSION TO humans BELAY STANFORD

http://www.pnas.org/cgi/content/full/99/suppl_4/16378

It appears quite likely that sheep scrapie spreads by ingestion of the infectious agent, although the source has not
been established; infected placenta has been suggested (41), but scrapie-prion-contaminated feces are a likely
possibility that merits investigation

http://lib.bioinfo.pl/auth:Will,RG

Vet Immunol Immunopathol. 2007 Feb 2; : 17320972       
       
Quantification of Peyer's patches in Cheviot sheep for future scrapie pathogenesis studies.            
Suzanne G St Rose , Nora Hunter , James D Foster , Dawn Drummond , Calum McKenzie , David Parnham , Robert G
Will , Mark E J Woolhouse , Susan M Rhind            
Peyer's patches (PPs) are the most probable sites of intestinal uptake of the transmissible spongiform encephalopathy
(TSE) agent.          

**************************************************************


http://www.jneurosci.org/cgi/content/abstract/24/50/11280

Ravi Shankar Mishra, * Subhabrata Basu, * Yaping Gu, Xiu Luo, Wen-Quan Zou, Richa Mishra, Ruliang Li, Shu G.
Chen, Pierluigi Gambetti, Hisashi Fujioka, and Neena Singh

Because ferritin shares considerable homology across species, these data suggest that PrPSc-associated proteins, in
particular ferritin, may facilitate PrPSc uptake in the intestine from distant species, leading to a carrier state in humans.
*********************************************************************
http://www.pnas.org/cgi/content/abstract/102/3/640

Although this global architecture coincides with the previously reported murine, Syrian hamster, bovine, and human
PrPC structures, there are local differences between the globular domains of the different species. Because the five
newly determined PrPC structures originate from species with widely different transmissible spongiform encephalopathy
records, the present data indicate previously uncharacterized possible correlations between local features in PrPC
three-dimensional structures and susceptibility of different mammalian species to transmissible spongiform
encephalopathies.

**************************************************************************************
Environmental sources of prion transmission in mule deer - group of 8 »
MW Miller, ES Williams, NT Hobbs, LL Wolfe - Emerg Infect Dis, 2004 - cdc.gov
... However, CWD infection–specific protease-resistant prion protein (PrP CWD ) accumulates
in gut ... alimentary shedding of the CWD agent in both feces and saliva ...
Cited by 9 - Cached - Web Search

Transmission of Prions
EP Transmission - The Journal of Infectious Diseases, 2002 - journals.uchicago.edu
... has not been established, infected placenta has been suggested [72]; however, scrapie
prion-contaminated feces are a possibility that merits investigation. ...
***************************************************
Bovine Spongiform Encephalopathy, Chronic Wasting Disease, Scrapie, and the Threat to Humans from … -
group of 4 »
PJ Bosque - CURRENT NEUROLOGY AND NEUROSCIENCE REPORTS, 2002 - current-reports.com
... Lateral transmission certainly occurs, but the vector or fomite involved is a mystery.
Saliva, urine, feces, or blood all might carry infectious prions. ...
Cited by 7 - Web Search - BL Direct
Chronic Wasting Disease and the Science in support of the Ban on Baiting and Feeding Deer. - group of 2 »
TR Van Deelen - 2003 - gwf.org
... diseased deer infect other deer) · Deer can get CWD by ingesting something contaminated
with the disease prion · CWD prions may be shed in feces and saliva ...

http://dnr.wi.gov/org/land/wildlife/whealth/issues/CWD/cwd-report.pdf

WISCONSIN
page 16 of 22: "Baiting and feeding artificially concentrates deer and facilitates increased animal to animal contact and
exposure to CWD-containing excreta. A consequence of increased contacts is an increased risk of disease
transmission among deer."
*************************************
http://biology.plosjournals.org/perlserv/plosonline/?request=get-document&doi=10.1371/journal.pbio.0020121
peer reviewed open access magazine

Chronic Wasting Disease—Prion Disease in the Wild

Steve Bunk
Steve Bunk is a freelance writer based in Boise, Idaho, United States of America. E-mail: stevebunk@sbcglobal.net
Feces is also a possible reservoir because animals nose in the ground for feed, and urine is yet another candidate,
because it is involved in the scenting activities of cervids.

Soil could be an environmental reservoir, because cervids ingest dirt to supplement their diets with minerals. Bucks
also lick soil on which does have urinated to ascertain their mating status. University of Wisconsin soil science
professor Joel Pedersen has discovered that abnormally folded prions stick to the surface of some soil types, such as
clay, resisting environmental and chemical damage. “Captive elk contracted CWD when introduced into paddocks
occupied by infected elk more than 12 months earlier, despite fairly extensive efforts to disinfect the enclosures,”
Pedersen notes. He has begun a five-year project to characterize interactions between infectious prions and soil
particles and determine the extent to which infectivity is retained.

******************************************************************************
http://www.stanford.edu/group/virus/prion/2005priya/newfindings.html

Prions may be transmitted indirectly from the environment

Miller et al. 2004. Environmental Sources of Prion Transmission in Mule Deer. Emerging Infectious Diseases www.cdc.
gov/eid Vol. 10, No. 6, June 2004

Chronic wasting disease, a prion disease affecting cervids, can be transmitted to susceptible animals indirectly, from
environments contaminated by excreta or decomposed carcasses. Under experimental conditions, mule deer
(Odocoileus hemionus) became infected in two of three paddocks containing naturally infected deer, in two of three
paddocks where infected deer carcasses had decomposed 1.8 years earlier, and in one of three paddocks where
infected deer had last resided 2.2 years earlier. Indirect transmission and environmental persistence of infectious
prions will complicate efforts to control CWD and perhaps other animal prion diseases.

***************************************************************************************
http://infectioncontrol.ucsfmedicalcenter.org/Infection_Control_Manual/Sec4-11_Human_Prion_Disease_2005.pdf
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO --
PATIENTS WITH CREUTZFELDT JAKOB DISEASE
PAGE 3 OF 21
LIST OF TISSUES WITH MEDIUM OR LOW INFECTIVITY -  Adipose tissue -- Adrenal gland -- blood -- bone marrow -
CSF -- dental pulp -- FECES -- gingival tissue -- heart -- INTESTINES -- kidney, etc.
***************************************************************************
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15893117&dopt=Citation

Med Hypotheses. 2005;65(1):47-54.        Related Articles,   Links          

Risk analysis of ectoparasites acting as vectors for chronic wasting disease.

Lupi O.
The mode of contamination is not known, although direct contact between infected and non-infected animals via saliva,
urine and feces have been considered. Increasing spread of CWD has raised concerns about the potential
transmission to humans and the conversion of human prion protein by CWD-associated prions has been demonstrated
in laboratory experiments
******************************************************************************************************
http://www.stanford.edu/group/virus/prion/2005priya/newfindings.html

Could other animal TSEs pose a risk to humans?

Belay et al. 2004. Chronic Wasting Disease and Potential Transmission to Humans. Emerging Infectious Diseases Vol.
10, No. 6, June 2004, pg. 977-984.

Chronic wasting disease (CWD) of deer and elk is endemic in a tri-corner area of Colorado, Wyoming, and Nebraska,
and new foci of CWD have been detected in other parts of the United States. Although detection in some areas may be
related to increased surveillance, introduction of CWD due to translocation or natural migration of animals may account
for some new foci of infection. Increasing spread of CWD has raised concerns about the potential for increasing human
exposure to the CWD agent. The foodborne transmission of bovine spongiform encephalopathy to humans indicates
that the species barrier may not completely protect humans from animal prion diseases. Conversion of human prion
protein by CWD associated prions has been demonstrated in an in vitro cell free experiment, but limited investigations
have not identified strong evidence for CWD transmission to humans. More epidemiologic and laboratory studies are
needed to monitor the possibility of such transmissions.

Risk analysis of ectoparasites acting as vectors for chronic ...

... the potential transmission to humans and the conversion of human prion protein by CWD-associated prions has
been demonstrated in laboratory experiments. ...
www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve& db=PubMed&list_uids=15893117&dopt=Abstract - Similar pages
Although there is no evidence that CWD has been transmitted to humans, it may have the potential to infect humans.
Salman MD
  

RESEARCH

American Journal of Pathology. 2005;167:1033-1042.)
Bovine Prion Is Endocytosed by Human Enterocytes via the 37 kDa/67 kDa
Laminin Receptor Etienne Morel*, Thibault Andrieu, Fabrice Casagrande, Sabine Gauczynski,  Stefan Weiss, Jacques
Grassi, Monique Rousset*, Dominique Dormont and Jean  Chambaz*

From UMR505 INSERM/Université Pierre et Marie Curie,* Paris, France; the
Service de Neurovirologie, Commissariat à l'Energie Atomique (CEA),
Fontenay aux Roses, France; the Service de Pharmacologie et d'Immunologie
CEA/Saclay, Gif-sur-Yvette, France; and the Laboratorium für Molekulare
Biologie-Genzentrum, Institut für Biochemie der Ludwig-Maximilian
Universität München, Munich, Germany

”Some forms of transmissible spongiform encephalopathies result from oral  infection. We have thus analyzed the early
mechanisms that could account  for an uptake of infectious prion particles by enterocytes, the major cell population of
the intestinal epithelium. . . “. “We observed internalization by enterocytes of  prion particles from bovine spongiform
encephalopathy brain homogenates  but not from mouse-adapted scrapie-strain brain homogenates or purified  bovine
spongiform encephalopathy scrapie-associated fibrils. Bovine prion  particles were internalized via endocytosis within
minutes of infection
and were associated with subapical vesicular structures related to early  endosomes. “

“Altogether, our results underscore a potential role of enterocytes in the
absorption of bovine prions during oral infection through specific LRP/LR-
dependent endocytosis.”

http://ajp.amjpathol.org/cgi/content/abstract/167/4/1033

Enterocyte is a type of epithelial cell of the superficial layer of the small and large intestine tissue. These cells can help
break up molecules and transport them into the tissues.

Unexpectedly high incidence of visceral AA-amyloidosis in slaughtered cattle
in Japan . . . increase in CJD in Japan . . . .

Authors: Kana Tojo a; Takahiko Tokuda a; Yoshinobu Hoshii b; Xiaoying...eiichi Higuchi c; Takane Matsui d; Fuyuki
Kametani e; Dr Shu-Ichi Ikeda a Affiliations: a Third Department of Medicine, Shinshu University School of Medicine.
Matsumoto. Japan b First Department of Pathology, Yamaguchi University School of Medicine. Ube. Japan c
Department of Aging Biology, Institute on Aging and Adaptation, Shinshu University, Graduate School of Medicine.
Matsumoto. Japan d Department of Pathobiological Science, School of Veterinary Medicine,
Obihiro University of Agriculture and Veterinary Medicine. Obihiro. Japan e Department of Molecular Biology, Tokyo
Institute of Psychiatry. Tokyo. Japan

“Rather, it may be the true fact that in Japan our results reflect to a large
extent a genuine increase in CJD. The number of iCJD cases may still
increase even after the total ban on the practice of causal grafts.5,8
Regarding sporadic CJD (sCJD), a recent report from the European Unions collective study on CJD suggests that the
mortality rates from sCJD increased with time between 1993 and 2002.20 It is quite probable that this temporal
increase of sCJD may also exist in Japan. The increase may have
been accompanied to some extent by the improvement of physicians diagnostic”skills for CJD since 1997 when a
manual for clinical practice on CJD was introduced in our country.20,21


January 2004   Mad-Cow Worries
May Force Change   In Rules for Feed
By ANNA WILDE MATHEWS and SARAH LUECK Staff Reporters of THE WALL STREET JOURNAL WASHINGTON --
The Bush administration, under pressure to further reduce
the risk of mad-cow disease, is preparing to tighten rules on animal feed, as well as human food and dietary
supplements.

The moves by the Food and Drug Administration are likely to close several loopholes that might allow potentially
contaminated materials derived from cattle to be fed back to cattle and other livestock, according to people with
knowledge of the matter. The FDA would ban the use of cow blood and blood products and poultry litter, among
otheringredients, in feed for cows, these people say. The agency also is leaning toward banning the use of certain
high-risk material, such as cows' small intestines, in human food or supplements.

http://www.organicconsumers.org/madcow/mad12901.cfm

-- According to the World Health Organization, tainted protein feed made from carcasses of sick animals spread BSE in
Europe. The feed was sold all over the world, even after Britain imposed a domestic ban on it in 1988. Also, potentially
contaminated European meat products were imported to the U.S. well into the '90s.

Prions are a hardy lot, able to withstand radiation, temperatures of 600F [600 degrees Celsius, which is over a
thousand degrees Fahrenheit--], and standard hospital disinfection. In Europe, policies that underestimated the threat
have proven disastrous. ''It reminds me of the early days of HIV, when the blood industry wouldn't do anything,'' says
Michael Hansen, who analyzes U.S. prion disease policies for the Consumers Union. ''If you want to forestall disaster,
you have to take a precautionary approach,'' he says.
While bad feed is blamed for spreading BSE, scientists believe prions from CWD and the sheep disease scrapie CAN
BE SHED THROUGH MUCUS AND FECES, THUS INFECTING THE LAND.
***********************************************************
http://www.zyworld.com/Alan_Long/BSEin%20sheep.htm

BSE in Sheep - Report of the Core Stakeholder Group, May 2002
"1.7
Birds, vermin, wildlife and fish
1.7.1 These are all carriers, whether farmed or scavengers, of zoonotic diseases. They cannot be disregarded in
transmissions and passage of prions; nor can the disposal of EXCRETA, afterbirths, and mortal remains of all these
species, as well as of affected sheep and goats, be overlooked. Relevant interactions among food-producing animals,
wild and domesticated animals, and insect and parasitic vectors can be detected in zoonotic diseases such as
tuberculosis, paratuberculosis, Lyme disease, and liver fluke.

2 Transmissions other than by feeds

2.1 Grazing animals are farmed in conditions, some definitely ‘unnatural’, in which they are in contact with wild animals
and their EXCRETA, as well as with carriers of prion diseases among their own species. Environmental contamination
with infective particles (and prions are robust) has been little investigated, although VEGA has for many years drawn
attention to consequences that reach into water supplies and possible means of suspect transmissions. Modern
domestic farm animals graze over recently-soiled territories, and cattle and sheep may be grazed together. These
animals are kept in conditions that do not allow them much chance to reserve areas as LATRINES; and a grazing
animal can consume large amounts of soil with grass. The present problems arising with the association of badgers
and cattle in tuberculosis and of dogs and foxes and sheep with hydatidosis illustrate this point."

www.nrel.colostate.edu/projects/ cwd/papers/progress_report.pdf

"Because we believe that FECES is likely to be a major route of excretion of the CWD agent, mule deer fawns are
being hand raised at a non-CWD endemic facility for exposure to feces from the experimentally inoculated deer. This
will allow us to gather data on the potential for FECAL SHEDDING in advance of the results from the transgenic mice."


January 2004
Dear Ms. Shields,

Avenues of transmission of prions amongst captive and wild cervids (deer and elk) are not clearly understood.
Anecdotal evidence suggest that when animals are in large concentrations over time (captives) various modes of
transmission (including contact with EXCRETA) likely play a role. However, in the WILD, contact with EXCRETA is likely
not as efficient a way to maintain or spread the disease. Much of this information is
speculation. To date, prions have not been detected in saliva yet most researchers are reluctant to exclude SALIVA as
a source. While prions have not been detected inURINE other metabolites associated with prion disease seem to have
promise as a diagnostic tool. I am not aware of specific studies of feces to detect prions. However, because prions
have been detected in lymphoid tissue within the INTESTINES (Peyer's patches), FECES seems a good candidate as a
source.

The reason that studies have not been forthcoming on this matter is because of the difficulty in developing a reliable
test. So far, infectivity studies are based upon the ability of an unknown sample to either cause illness or death in an
animal.

Typically, urine and feces are toxic by themselves and so efforts to either
extract potentially harmful particles or neutralize the excreta are very
difficult making tests of these materials unreliable.


Scott D. Wright, Ph.D. Branch Chief, Disease Investigations  USGS National Wildlife Health Center   6006 Schroeder
Road   Madison, Wisconsin 53711  608-270-2460
608-270-2415 FAX
http://www.iol.ie/~craigie/BSE/Pages/Transfer.htm  PEYERS PATCHES

The first obstacle that the PRION has to surmount is the gastric juice which contains enzymes to destroy protein, but it
is known that this mechanism sometimes leaks allowing through between one in a million and one in ten million protein
molecules, this is accentuated during illness or after drinking alcohol.
The prion is then thought to LEAVE THE GUT via part of the immune system, the PEYERS PATCHES. These are small
nodules scattered over the inside of the intestines, with an outer layer of specialised cells that are very good at
absorbing bacteria, proteins and viruses from the gut. This is used by the body to detect organisms and molecules that
the immune system needs to be aware of. From here the route of the PRION is thought to lead it through the tissues
surrounding the GUT, and then into the peripheral nerves, through which it travels up to the brain; although this route
is not known for certain, and it is not known whether the body contains any special measures to protect against the
passage of PRION diseases. Only once the PRIONS reach the brain can the disease begin.
*********************************************************************
http://www.vegsource.com/talk/madcow/messages/9911604.html

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.
*************************************************
p-. 160
of Mad Cow - USA - "For that matter, FDA wasn't planning to ban whole sheep -- just the "specified offals"
known to contain the highest concentrations of infection, which FDA defined as "any tissue from the brain, spinal cord,
spleen, thymus, tonsil, lymph nodes, or INTESTINES (DUODENUM TO ANUS, inclusive)."
*****************************************************************************
http://www.hawgslimited.com/CWD_html/CWD1.htm

According to experts it’s become suspect CWD is spread through TSE prions, the mutant protein causing CWD. And
proteins (including prions) ARE PASSED IN BOTH URINE AND FECES. In an article appearing in the NATIONAL POST
Rob Miskosky, editor of Alberta Outdoorsmen, says, "Scientific journals confirm TSE prions have been found in URINE
long before the disease is detected in animals and COULD REMAIN VIABLE IN THE SOIL FOR YEARS."
*********************************************************************
http://www.unizh.
ch/pathol/neuropathologie/pdf/KleinM_AguzziA_2000_The_neuroimmune_interface_in_prion_disease___NewsPhysiolS
ci15_250-255.pdf

"For example, BSE appears to have low affinity for lymphoid tissue of cow and is confined to the nervous system of
experimentally infected cattle, although very limited amounts of infectivity have been detected in other sites, SUCH AS
TERMINAL ILEUM AND BONE MARROW. In contrast, during the human disease vCJD which is most probably derived
from BSE, PrP accumulates in the tonsils, spleen and appendix of infected individuals."
*************************************************************
CROSSING THE "SPECIES BARRIER"
A disease run wild; a killer of deer and elk breaks out of Colorado "We don't
know how it would jump," said Gregory Raymond, a federal scientist with
Rocky Mountain Laboratory of Persistent Viral Diseases in Hamilton, Mont.

"However, we do know it's possible for diseases of one species to cross the
barrier to other species."

When mad cow disease crossed the species barrier to kill humans, it became
known as new variant Creutzfeldt-Jakob disease, or vCJD.

"A number of people now recall that the sheep in the corrals at CSU had
scrapie, but no documentation has been found to prove it.

"There were a number of deer projects going on at the time and deer were
coming in from the wild that may have been infected, and we were trading
deer with Sybille (the Wyoming Game and Fish Department's Sybille Research
Unit, near Wheatland, Wyo.), and so it's impossible to say for sure how it
got started," Schoonveld said.

"But my guess as a biologist is those sheep had scrapie, and in close
confinement - something that they wouldn't do out in the wild - it jumped to
deer and infected them.

Colorado Division of Wildlife veterinarian Miller, Wyoming Game and Fish
veterinarian Thorne and others believe in the "lateral transfer" of the
disease, meaning it passes directly through FECES, URINE OR SALIVA. IT ALSO SEEMS LIKELY THAT IT
CONTAMINATES THE GROUND AND PLANTS, AND THEN REMAINS CONTRACTIBLE FOR MANY YEARS."
*********************************************************************
http://www.medical-library.org/journals2a/creutzfeldt_jacob2.htm

Pathogenesis
In lambs exposed to scrapie-infected flocks, infectivity is first found at about one year of age in the lymphatic tissues
and intestines, suggesting transmission by way of the alimentary tract. Infectivity in the brain is found at about two
years of age, and infectivity slowly increases in the brain, with resultant spongiform changes and clinical disease during
the

***************************************************************************
http://humrep.oupjournals.org/cgi/content/abstract/17/10/2501

Prion transmission in blood and urine: what are the implications for recombinant and urinary-derived
gonadotrophins?
H. Reichl1,4, A. Balen2 and C.A.M. Jansen3
1 Hämosan Life Science Services, Vienna Biocenter, Dr Bohr Gasse 7b, A-1030 Wien, Austria,  2 Leeds General
Infirmary, Leeds LS2 9NS, UK and
3 Reinier de graafgroep, loc Diaconessenhuis, Fonteynenburghlaan 5, 2275 CX Voorburg, The Netherlands
Evidence is emerging that suggests that the protease-resistant isoform (PrPsc) of the normal cellular prion protein
(PrPc) can be detected in the BLOOD AND URINE OF ANIMALS AND HUMANS with transmissible spongiform
encephalopathies (TSEs).
*********************************************************************
http://pubs.acs.org/cen/topstory/7929/7929notw7.html

NEWS OF THE WEEK
SCIENCE
July 16, 2001
Volume 79, Number 29

PROBE FOR PRIONS   DIAGNOSTIC PRIONS FOUND IN URINE BEFORE DISEASE SYMPTOMS APPEAR    STU
BORMAN
In a surprising and unexpected discovery, researchers in Israel have found a form of prion in URINE that can indicate
the presence of disease caused by the mysterious protein even before symptoms appear.   The findings suggest that a
urine test could be developed to identify cows with mad cow disease, people with CJD , sheep with scrapie, and animals
with other forms of prion diseases--transmissible spongiform encephalopathies, or TSEs.
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http://www.organicconsumers.org/madcow/early82101.cfm

In the research in Israel, Gabizon's team also tried to see whether the prion particles in urine are infectious, but no
cases of prion-induced disease were seen in the hamsters. But there were signs that the prion particles - perhaps
incomplete or altered prion particles - got into the animals' brains. So the Hadassah team concluded that "the clinical
and epidemiological implications" of its findings are yet to be determined.

Commenting on the Israeli team's work, neuroscientist Huntington Potter, of the University of South Florida College of
Medicine, said, "The finding of scrapie-like protein in the urine before clinical symptoms is going to be very helpful for
both the veterinary and medical diagnosis of these diseases." But he also questioned whether prion-based diseases
can be transmitted via urine. The Israeli researchers, he said, gave "no data addressing that question." And he called
their raising the "alarming possibility" of such transmission a "hyper-scare paragraph" that he found confusing.
Nonetheless, Potter said, this idea of disease transmissibility via URINE "is definitely worth following up. It has been a
MYSTERY IN SCRAPIE about how the animals infect each other in the field, and this is as good a hypothesis as any."
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http://www.organicconsumers.org/madcow/urine71701.cf

If the rogue proteins are in lymph nodes, Dr. Gabizon said, it stands to reason that they will end up in the bloodstream.
Because the abnormal proteins cannot be broken down by enzymes in blood, she said, it also stands to reason that
they will be cleared from the body by the kidneys.

The rogue proteins are small enough to pass through the kidneys' filtration system, she said, so they should turn up in
urine.

To find the rogue proteins in urine, Dr. Gabizon and her assistant, Gideon Shaked, came up with a purification scheme.
The kidneys contain urea, a compound that disrupts protein folding but does not destroy the proteins. The researchers
suspected that the rogue proteins might be altered by urea and excreted in urine as particles that would not be
recognized as prions.

To test that idea, the researchers took urine samples from hamsters, humans and cattle infected with known prion
diseases, and from healthy controls. The urine was put into a machine that removes urea, a process that allows
proteins to fold back into their original shapes. These refolded proteins were then exposed to enzymes that broke down
normal proteins, but not prions.

In healthy controls, all proteins were destroyed. But in animals and people with prion diseases, one protein could not
be broken down. Dr. Gabizon called that enzyme-resistant protein a URINARY PRION and said it was new. The
presence of such a protein is the diagnostic hallmark of prion diseases, she said.

Tests show that the urinary prion is not overtly infectious, although it may have some low level of infectivity, Dr.
Gabizon said. Hamsters injected with the particles have not developed any signs of disease after 300 days but are still
under observation. Other rodents have been shown to harbor infectious proteins without showing signs of disease, she
said, and yet as carriers they can pass on the disease.

The urinary prion seems to be an excellent marker for the progression of prion diseases, Dr. Gabizon said. For
example, hamsters were injected with the classic rogue prion that causes scrapie, a sheep disease. For one week, the
hamsters excreted the urinary prion, then stopped. Sixty days later, the urinary prion reappeared in increasing
amounts. Dr. Gabizon said this suggested that as the infection built up in the body and brain, the amount of urinary
prion increased.     "It is as if the urine was a mirror to the brain," she said.
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http://w3.aces.uiuc.edu/AnSci/BSE/BSE_Urine.htm

PrPRes in TSE Urine
The first report a protease resistant form of PrP in the urine of animals and humans affected with TSE is quite notable.
PrPRes was found in the urine of: hamsters inoculated with Scrapie as early as 17d post inoculation (long before
clinical and pathological changes) (however, not between 35 and 49d post inoculation)  these findings may indicate
clearance of inoculated PrPSc at day 17 in increasing amount from 56d post inoculation until clinical signs appeared
(day 105) cattle with BSE humans with CJD E200K mutation PrPRes was not found in the urine of:
normal hamsters, normal cattle , normal humans
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"Genesis Bioventures, Inc. (AMEX:GBI - formerly BioLabs, Inc.) announced today that one of its portfolio companies,
Prion Developmental Laboratories, Inc. (PDL) is currently in late-stage discussions with a European pharmaceutical
company to provide them with an easy-to-use, patents pending rapid strip test for detecting Bovine Spongiform
Encephalopathy (BSE), or Mad Cow Disease. " (in urine)
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http://www.bmu.de/english/download/soil/files/bse_tse.pdf
ON PAGE 19

"The leaching of prions, for examples, from landfills, (this concerns landfills of BSE-infected animal corpses) has
likewise been recognized and risk-reducing measures recommended (SSC, 1999). Possible methods of assessing the
persistence of the infectivity in soil, e.g. in soil of grazing land must be investigated."

(NOTE: In NH and many other parts of the country, landfill leachates are discharged into the local POTWs !! -- PRIONS
ARE NEARLY INDESTRUCTIBLE -- REQUIRING TEMPERATURES EXCEEDING 212 DEGREES FAHRENHEIT . . . In
August 2002 the Wisconsin Dept. of Natural Resources published a Risk Assessment acknowledging that -- with regard
to landfill leachate containing prions from CWD infected deer --the prions will survive the wastewater treatment process
and be concentrated in the sewage sludge which is spread on agricultural fields and cattle grazing lands throughout
the Midwest and Rocky Mountain states . . . . some of which are areas where CWD is endemic.)

-- Page 16 - "As long as animals are exposed to infected feed, the elimination of some BSE infectivity with faeces is
likely to take place. This will be the case for both BSE-susceptible and non-BSE=susceptible species. The extent of this
elimination depends on the amount of BSE infectivity ingested during the period of exposure as well as on the species
involved and the health conditions of the involved animals."

Page 7 - "With respect to the entry route via farm manure, there is consensus that ingested infected material is partially
degraded or inactivated metabolically but is also EXCRETED UNCHANGED and thus enters the soil."

Page 8 - "The same applies, for example, for the entry route via SEWAGE SLUDGE. Although it can be assumed that
the residual infectivity or the content of pathogenic prion proteins is very low in sewage sludge, the possibility of this
route, e.g. for sewage sludge from waste water from rendering plants, cannot be ruled out, as long as there is no
guarantee that no infected animals were processed."

(We're talking about AT LEAST 100,000 "Downers cows" each year in the USA -- poor cows which can't get
up - or which are so-called "Rabies negative" cows -- or cows exhibiting CNS symptoms) .... which are
slaughtered and rendered each year -- many of which could be infected with prion disease - transmissible
spongiform encephalopathy (TSE).
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http://www.veggieplace.com/printarticle.asp?ID=228

PRIONS IN MUSCLE TISSUE?

"For years, the prions were thought to be confined mostly to the brain and spinal cord. The research could mean that
scientists must look further. Prusiner has called for the testing of muscles in infected deer, elk and cattle. (Prusiner was
considered to be mad until he won the Nobel Prize in medicine in 1997 for his research on prions.)
Simply put, muscle is meat, the part consumed by humans.
Says Race: "No one knows the full range of the human implications."
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- [ ]
http://www.www.petrinum.ac.at/schule/buch/framepg/bse2.htm


... Beim Menschen wurden bei der Creutzfeldt-Jakob-Krankheit, die über Rinder übertragene
nvCreutzfeld-Jakob ... icon4.gif (3061 bytes). Creutzfeldt-Jakob-Disease ..
.
www.petrinum.ac.at/schule/buch/framepg/bse2.htm - 30k - (Search term: nCreutzfeld Jakob Disease"

"BSE is an illness, which arises actually only with cattle. Since it is transferable on humans however from the cattle, and
there to one Creutzfeldt Jakob illness similar illness, which n.v. Creutzfeldt Jakob illness (n.v.=new variant) leads, is
explained in detail it here"

With humans with the Creutzfeldt Jakob illness, the nvCreutzfeld Jakob illness, transferred over cattle, Kuru, the
Gerstmann Straeussler syndrome and that fatal Familial Insomnia (FFI) were found to Prionen. There these changed
proteins (Prionen) all these illnesses to reason lie, summarize one them recently under the term Prionenkrankheiten .
The neurologist Stanley Prusiner received 1997 for the list of a theory about the nature of the Prionenkrankheiten to
the Nobelpreis for medicine.

Until November of the yearly 2000 88 humans and the 5 in France at the new illness nv. Creutzfeld Jakobs D deceased
in England.

Transmission paths

High concentrations are in the nerve tissue, also in the Lymphorganen like the spleen, to the placenta, where boy can
be infected, in the muscle and in the blood. The infection was originally probably spread over animal fodder by
admixture of animal flour.
Also by milk spare products, which contain animal fats, an infection could take place by BSE.
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http://organicconsumers.org/madcow/level6802.cfm

But wait a minute. As the Rocky Mountain News special report, "Killer in the Herds," pointed out last Saturday, recent
research is "raising alarms" about whether prions lodge in muscle tissue, too. Scientists have in fact detected high
levels of prions in the muscles of laboratory mice infected with transmissible spongiform encephalopathies, the family of
diseases that include CWD. As a result, Nobel laureate Stanley Prusiner is calling for the testing of muscles in infected
deer and elk as well.
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a lot more information remains to be consolidated into my computer on this subject . . . . ..













comment: If there are prions in urine -- then there are prions in the sewage sludge being spread on dairy farms and
cattle grazing lands across the country . . .. particularly in the mid west . . .where CWD is emdemic . . . . . .