Seeded amplification of chronic wasting disease prions in nasal brushings and recto-anal mucosa associated lymphoid tissues from elk by real time quaking-induced conversion
Seeded amplification of chronic wasting disease prions in nasal brushings
and recto-anal mucosa associated lymphoid tissues from elk by real time
quaking-induced conversion
Nicholas J. Haley#,a, Chris Siepkera, Laura L. Hoon-Hanksb, Gordon
Mitchellc, W. David Walterd, Matteo Mancae, Ryan J. Monellof, Jenny G. Powersf,
Margaret A. Wildf, Edward A. Hooverb, Byron Caugheye and Jürgen A. Richta +
Author Affiliations
Department of Diagnostic Medicine and Pathobiology, College of Veterinary
Medicine, Kansas State University (KSU), Manhattan, KS, USAa Department of
Microbiology, Immunology and Pathology, College of Veterinary Medicine and
Biomedical Sciences, Colorado State University, Fort Collins, CO, USAb Canadian
Food Inspection Agency, National and OIE Reference Laboratory for Scrapie and
CWD, Ottawa Laboratory Fallowfield, Ottawa, ON, Canadac U.S. Geological Survey,
Pennsylvania Cooperative Fish and Wildlife Research Unit, The Pennsylvania State
University, University Park, PA, USAd TSE/Prion Biochemistry Section, Laboratory
of Persistent Viral Diseases, Rocky Mountain Laboratories (RML), National
Institute of Allergy and Infectious Diseases, Hamilton, MT, USAe National Park
Service, Biological Resources Division, 1201 Oak Ridge Drive, Suite 200, Fort
Collins, Colorado 80525, USAf
ABSTRACT
Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of
cervids, was first documented nearly fifty years ago in Colorado and Wyoming,
and has since been detected across North America and to the Republic of Korea.
The expansion of this disease makes the development of sensitive diagnostic
assays and antemortem sampling techniques crucial for the mitigation of spread;
this is especially true in cases of relocation/reintroduction, or prevalence
studies in large or protected herds where depopulation may be contraindicated.
This study sought to evaluate the sensitivity of the real-time quaking-induced
conversion (RT-QuIC) assay in recto-anal mucosa associated lymphoid tissue
(RAMALT) biopsies and nasal brushings collected antemortem. These findings were
compared to results from ante- and postmortem samples evaluated using
immunohistochemistry (IHC). RAMALT samples were collected from populations of
farmed and free-ranging Rocky Mountain elk (Cervus elaphus nelsoni, n=323), with
nasal brushes collected from a subpopulation of these animals (n=205). We
hypothesized the sensitivity of RT-QuIC would be comparable to IHC in RAMALT,
and would correspond to IHC of postmortem tissues. We found RAMALT sensitivity
(77.3%) to be highly correlative between RT-QuIC and IHC. Sensitivity was lower
when testing nasal brushings (34%), though both RAMALT and nasal brush
sensitivities were dependent on both PRNP genotype and disease progression
determined by obex score. These data suggest that RT-QuIC, like IHC, is a
relatively sensitive assay for detection of CWD prions in RAMALT biopsies, and
with further investigation has potential for large scale and rapid automated
testing for CWD in antemortem samples.
FOOTNOTES
↵#Corresponding author (e-mail: nicholas.j.haley@gmail.com) Copyright ©
2016, American Society for Microbiology. All Rights Reserved. http://jcm.asm.org/content/early/2016/02/11/JCM.02700-15.abstract
Sunday, February 14, 2016
Antemortem detection of chronic wasting disease prions in nasal brush
collections and rectal biopsies from white-tailed deer by real time
quaking-induced conversion
Real and perceived issues involving animal proteins C. R. Hamilton May 3,
2002, a review of USDA MAD COW DISEASE BSE FEED
FIRST, please see the drastic decrease of mad cow disease in the UK after
the mad cow feed ban went into force ;
SUMMARY OF PASSIVE SURVEILLANCE REPORTS IN GREAT BRITAIN
PASSIVE SURVEILLANCE
SUSPECTS RESTRICTIONS LIFTED DUE TO ALTERNATIVE SLAUGHTER NOT YET SUSPECTS
KNOWN TO HAVE BEEN SLAUGHTERED SUSPECTS WITH OUTCOME SLAUGHTERED SUSPECTS IN
WHICH BSE NOT SLAUGHTERED SUSPECTS IN WHICH BSE PRIVATE SUBMISSIONS IN WHICH
YEAR RESTRICTED DIAGNOSIS NOTIFIED SLAUGHTERED PENDING CONFIRMED CONFIRMED
BSE CONFIRMED
Pre 18.07.88 1142 1 0 980 0 414 727 0
1988 2512 140 0 2372 0 192 2180 4
1989 8443 386 0 8057 0 924 7133 4
1990 17323 682 0 16641 0 2460 14181 0
1991 30003 982 0 29021 0 3995 25026 6
1992 44844 1690 0 43154 0 6474 36680 2
1993 42931 1847 0 41084 0 6714 34370 0
1994 30259 1551 0 28708 0 4765 23943 2
1995 17945 789 0 17156 0 2855 14301 1
1996 10697 547 0 10150 0 2137 8013 3
1997 5604 302 0 5302 0 992 4310 3
1998 4291 260 0 4031 0 852 3179 1
1999 2984 146 0 2838 0 582 2256 2
2000 1870 117 0 1753 0 442 1311 0
2001 1218 66 0 1152 0 371 781 0
2002 877 68 0 809 0 364 445 0
2003 456 43 0 413 0 240 173 0
2004 351 41 0 310 0 228 82 0
2005 186 30 0 156 0 117 39 0
2006 144 26 0 118 0 103 15 0
2007 69 11 0 58 0 51 7 0
2008 39 7 0 32 0 30 2 0
2009 22 7 0 15 0 14 1 0
2010 19 7 0 12 0 12 0 0
2011 11 2 0 9 0 9 0 0
2012 5 2 0 3 0 3 0 0
2013 2 0 0 2 0 2 0 0
2014 2 0 0 2 0 2 0 0
2015 0 0 0 0 0 0 0 0
2016 0 0 0 0 0 0 0 0
Total 223107 9749 0 213358 0 34930 178428 28
Data valid to 31 January 2016
SECOND, what has MAFF...now DEFRA, what have they said about the mad cow
feed ban in the USA in 2012?
Friday, December 14, 2012
DEFRA U.K. What is the risk of Chronic Wasting Disease CWD being introduced
into Great Britain? A Qualitative Risk Assessment October 2012
snip...
In the USA, under the Food and Drug Administration’s BSE Feed Regulation
(21 CFR 589.2000) most material (exceptions include milk, tallow, and gelatin)
from deer and elk is prohibited for use in feed for ruminant animals. With
regards to feed for non-ruminant animals, under FDA law, CWD positive deer may
not be used for any animal feed or feed ingredients. For elk and deer considered
at high risk for CWD, the FDA recommends that these animals do not enter the
animal feed system. However, this recommendation is guidance and not a
requirement by law.
Animals considered at high risk for CWD include:
1) animals from areas declared to be endemic for CWD and/or to be CWD
eradication zones and
2) deer and elk that at some time during the 60-month period prior to
slaughter were in a captive herd that contained a CWD-positive animal.
Therefore, in the USA, materials from cervids other than CWD positive
animals may be used in animal feed and feed ingredients for non-ruminants.
The amount of animal PAP that is of deer and/or elk origin imported from
the USA to GB can not be determined, however, as it is not specified in TRACES.
It may constitute a small percentage of the 8412 kilos of non-fish origin
processed animal proteins that were imported from US into GB in 2011.
Overall, therefore, it is considered there is a __greater than negligible
risk___ that (nonruminant) animal feed and pet food containing deer and/or elk
protein is imported into GB.
There is uncertainty associated with this estimate given the lack of data
on the amount of deer and/or elk protein possibly being imported in these
products.
snip...
36% in 2007 (Almberg et al., 2011). In such areas, population declines of
deer of up to 30 to 50% have been observed (Almberg et al., 2011). In areas of
Colorado, the prevalence can be as high as 30% (EFSA, 2011). The clinical signs
of CWD in affected adults are weight loss and behavioural changes that can span
weeks or months (Williams, 2005). In addition, signs might include excessive
salivation, behavioural alterations including a fixed stare and changes in
interaction with other animals in the herd, and an altered stance (Williams,
2005). These signs are indistinguishable from cervids experimentally infected
with bovine spongiform encephalopathy (BSE). Given this, if CWD was to be
introduced into countries with BSE such as GB, for example, infected deer
populations would need to be tested to differentiate if they were infected with
CWD or BSE to minimise the risk of BSE entering the human food-chain via
affected venison.
snip...
The rate of transmission of CWD has been reported to be as high as 30% and
can approach 100% among captive animals in endemic areas (Safar et al.,
2008).
snip...
In summary, in endemic areas, there is a medium probability that the soil
and surrounding environment is contaminated with CWD prions and in a
bioavailable form. In rural areas where CWD has not been reported and deer are
present, there is a greater than negligible risk the soil is contaminated with
CWD prion.
snip...
In summary, given the volume of tourists, hunters and servicemen moving
between GB and North America, the probability of at least one person travelling
to/from a CWD affected area and, in doing so, contaminating their clothing,
footwear and/or equipment prior to arriving in GB is greater than negligible.
For deer hunters, specifically, the risk is likely to be greater given the
increased contact with deer and their environment. However, there is significant
uncertainty associated with these estimates.
snip...
Therefore, it is considered that farmed and park deer may have a higher
probability of exposure to CWD transferred to the environment than wild deer
given the restricted habitat range and higher frequency of contact with tourists
and returning GB residents.
snip...
THIRD, THE USDA FDA TRIPLE MAD COW DISEASE FIREWALL, WAS NOTHING MORE THAN
INK ON PAPER !
now, let’s just for a moment put away the corporate junk science, and let’s
look at recent updated BSE, CWD, Scrapie, TSE Prion sound science, could not
hurt...
Saturday, January 31, 2015
European red deer (Cervus elaphus elaphus) are susceptible to Bovine
Spongiform Encephalopathy BSE by Oral Alimentary route
I strenuously once again urge the FDA and its industry constituents, to
make it MANDATORY that all ruminant feed be banned to all ruminants, and this
should include all cervids as soon as possible for the following
reasons...
======
In the USA, under the Food and Drug Administrations BSE Feed Regulation (21
CFR 589.2000) most material (exceptions include milk, tallow, and gelatin) from
deer and elk is prohibited for use in feed for ruminant animals. With regards to
feed for non-ruminant animals, under FDA law, CWD positive deer may not be used
for any animal feed or feed ingredients. For elk and deer considered at high
risk for CWD, the FDA recommends that these animals do not enter the animal feed
system.
***However, this recommendation is guidance and not a requirement by law.
======
31 Jan 2015 at 20:14 GMT
*** Ruminant feed ban for cervids in the United States? ***
31 Jan 2015 at 20:14 GMT
see Singeltary comment ;
Oral transmission and early lymphoid tropism of chronic wasting disease
PrPres in mule deer fawns (Odocoileus hemionus )
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...
Title: Experimental oral transmission of chronic wasting disease (CWD) to
red deer (Cervus elaphus elaphus): early detection and late stage distribution
of protease-resistant protein (PrP-res)
In this study, red deer (Cervus elaphus elaphus) were exposed to the prion
agent by oral administration of brain homogenates from infected Rocky Mountain
elk. Antemortem testing was performed at 7 months post infection and the deer
were euthanized when clinical disease was observed at approximately 18 months
after infection. The abnormal prion protein was assayed by immunohistochemistry,
enzyme linked immunosorbent assay and western blot. Abnormal prion protein was
found in the spinal cord, brainstem, cerebellum, midbrain, thalamus, and
cerebrum in all 4 infected red deer. Most of the lymph nodes throughout the body
were positive for abnormal prion proteins. Abnromal prion protein was observed
in some additional peripheral tissues in some but not all of the deer. In
particular, most areas of the gastrointestinal tract were positive for abnormal
prions, although the salivary glands were rarely positive. This study
demonstrates the potential for oral transmission of chronic wasting disease to
red deer and confirms the usefulness of the current testing methods for post
mortem diagnosis of the disease in this species.
*** Singeltary reply ; Molecular, Biochemical and Genetic Characteristics
of BSE in Canada Singeltary reply ;
SNIP...SEE FULL TEXT AND MORE HERE ;
Tuesday, February 16, 2016
Real and perceived issues involving animal proteins C. R. Hamilton May 3,
2002, a review of USDA MAD COW DISEASE BSE FEED AND CERVIDS
Terry S. Singeltary Sr.
posted by Terry S. Singeltary Sr. at
4:11 PM
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