Land Spreading of the TSE Prion Disease, blood tank for feed, plants,
vegetables, and sludge, stupid is as stupid does
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Grass Plants Bind, Retain, Uptake, and Transport Infectious Prions
Sandra Pritzkow, Rodrigo Morales , Fabio Moda 3, Uffaf Khan, Glenn C.
Telling, Edward Hoover, Claudio Soto
correspondence
email 3Present address: IRCCS Foundation Carlo Besta Neurological
Institute, 20133 Milan, Italy
Publication stage: In Press Corrected Proof
Open Access
Highlights
•Grass plants bind prions from contaminated brain and excreta •Prions from
different strains and species remain bound to living plants •Hamsters fed with
prion-contaminated plant samples develop prion disease •Stems and leaves from
grass plants grown in infected soil contain prions
Summary
Prions are the protein-based infectious agents responsible for prion
diseases. Environmental prion contamination has been implicated in disease
transmission. Here, we analyzed the binding and retention of infectious prion
protein (PrPSc) to plants. Small quantities of PrPSc contained in diluted brain
homogenate or in excretory materials (urine and feces) can bind to wheat grass
roots and leaves. Wild-type hamsters were efficiently infected by ingestion of
prion-contaminated plants. The prion-plant interaction occurs with prions from
diverse origins, including chronic wasting disease. Furthermore, leaves
contaminated by spraying with a prion-containing preparation retained PrPSc for
several weeks in the living plant. Finally, plants can uptake prions from
contaminated soil and transport them to aerial parts of the plant (stem and
leaves). These findings demonstrate that plants can efficiently bind infectious
prions and act as carriers of infectivity, suggesting a possible role of
environmental prion contamination in the horizontal transmission of the disease.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Received: October 31, 2014; Received in revised form: February 4, 2015;
Accepted: April 15, 2015; Published Online: May 14, 2015
© 2015 The Authors. Published by Elsevier Inc.
snip...
RESULTS
Prions Bind to Plants and Bound-PrPSc Efficiently Sustain Prion
Replication
To study whether plants can interact with prions, we exposed wheat grass
roots and leaves to brain homogenate from hamsters that have succumbed to prion
disease induced by experimental inoculation with the 263K prion strain. The
presence of PrPSc and infectivity attached to the plants was studied in vitro
using the protein misfolding cyclic amplification (PMCA) technique and in vivo
by infectivity bioassays. For in vitro analyses, the plant tissues (roots and
leaves) were incubated for 16 hr with serial dilutions of 263K-brain homogenate
ranging from 10 1 to 10 8. Roots and leaves were washed thoroughly and analyzed
for the presence of PrPSc by serial PMCA (Morales et al., 2012). The results
show that even highly diluted PrPSc can bind to roots and leaves and sustain
PrPC conversion (Figure 1A). Although a direct comparison cannot be made,
because of differences on the effective surface, roots appear to retain PrPSc
better than leaves. However, both roots and leaves capture PrPSc efficiently,
even at very small concentrations, equivalent to those present in biological
fluids, such as blood and urine (Chen et al., 2010). By comparing the detection
of PrPSc-bound to plants (Figure 1A) with an experiment in which the same
dilutions of 263K brain homogenate were added directly to the tubes containing
normal brain homogenate and an equivalent piece of leaves or roots (Figure 1B),
we can estimate that a high proportion of PrPSc present in the sample was
attached to the plant tissue. Importantly, no detection of PrPSc was observed
when leaves and roots were exposed to normal brain homogenate (Figure 1C).
However, comparing PMCA amplification in the presence (Figure 1B) or in the
absence (Figure S1A) of plant tissue, it is possible to appreciate that plants
(both leaves and roots) partially inhibits the PMCA reaction. This explains why
in most of the experiments with plants, protease-resistant PrPSc is only
observed after two rounds of PMCA. In our current PMCA settings, no
false-positive PrPSc signals were ever detectable when samples did not contain
PrPSc inoculum (Figure S1B). These results indicate that leaves and roots can
efficiently bind PrPSc, which remains able to catalyze PrPC to PrPSc conversion,
leading to prion replication. In these experiments, plant tissues were incubated
with prions for 16 hr, but a similar experiment in which roots and leaves were
exposed to a 10 5 dilution of 263K brain homogenate for different times, we
found that as little as 2 min of incubation was sufficient for the efficient
contamination of plants (Figure S2).
Animals Can Be Infected by Oral Administration of Prion-Contaminated
Plants
To investigate whether prion-contaminated plants were able to infect
animals by ingestion, leaves and roots previously incubated with either
263K-infected or control hamster brain homogenates were orally administered into
naive hamsters. After exposure, plants were extensively washed five times with
water and animals were fed with dried material. As positive controls, we orally
administered 750 ml of 5% 263K brain homogenate (same material used to
contaminate plant tissue). All animals that ingested prion contaminated leaves
and roots developed typical prion disease. Although the incubation times were
significantly longer in animals ingesting prions attached to leaves and roots as
compared with those fed directly with the brain material, the differences were
not as high as one could have expected (Figure 2A). Indeed, incubation periods
were 147 ± 10,
159 ± 10, and 164 ± 13 days (mean ± SEM) for the groups inoculated with
brain homogenate, and prion contaminated roots and leaves, respectively. Prion
disease was confirmed by histological study of PrPSc deposition, astrogliosis,
and brain vacuolation (Figure 2B), as well as by biochemical detection of
protease-resistant PrPSc by western blot (Figure 2C). None of the animals
inoculated with leaves and roots exposed to normal brain homogenate developed
disease up to 550 days post-inoculation. Histological analysis did not show any
PrPSc staining or disease specific alteration in control animals.
Plants Bind Prions from Different Strains and Species To analyze
prion-plant interaction with other species and strains of the prion agent, we
performed similar studies as described in Figure 1, by incubating leaves and
roots with a preparation containing hamster, murine, cervid, and human prions
corresponding to the Hyper, 301C, CWD, and vCJD prion strains, respectively.
PrPSc from these strains and species showed good amplification by PMCA, using
homologous substrates (Figure S3A). In all cases, leaves and roots bound prions
from these species and retained the ability to replicate in vitro (Figure S3B),
indicating that the interaction of PrPSc with plants is a general feature of
infectious prions.
Contamination of Plants with Prions Excreted in Urine and Feces
Under natural conditions, it is likely that the main source of prions in
the environment comes from secretory and excretory fluids, such as saliva,
urine, and feces. We and others have shown that PrPSc is released in these
fluids and excretions in various animal species (Gonzalez-Romero et al., 2008;
Haley et al., 2009, 2011; Maddison et al., 2010; Terry et al., 2011; Moda et
al., 2014). It has been estimated that the amount of infectious prions spread by
excreta during the animals’ lifespan could match or even surpass the quantity
present in the brain of a symptomatic individual (Tamgu¨ ney et al., 2009). To
study whether plant tissue can be contaminated by waste products excreted from
prion-infected hamsters and deer, leaves and roots were incubated with samples
of urine and feces and the presence of PrPSc analyzed by serial rounds of PMCA.
For these experiments, plant tissues were incubated for 1 hr with urine or feces
homogenates obtained either from 263K-infected hamsters or CWD-affected cervids.
This time was chosen because longer incubation with these biological fluids
affected the integrity of the plant tissue. After being thoroughly washed and
dried, PrPSc attached to leaves and roots was detected by PMCA. The results
clearly show that PrPSc was readily detectable after three or four rounds of
PMCA in samples of wheat grass leaves and roots exposed to both urine and feces
from 263K sick hamsters (Figure 3A) and CWD-affected cervids (Figure 3B).
Comparing these results with studies of the direct detection of PrPSc in urine
and feces (Figures 3A and 3B), it seems that the majority of PrPSc present in
these waste products was effectively attached to leaves and roots. No signal was
observed in plant tissue exposed to urine or feces coming from non-infected
hamsters.
Prions Bind to Living Plants
To investigate a more natural scenario for prion contamination of living
plants, we sprayed the leaves of wheat grass with a preparation containing 1%
263K hamster brain homogenate. Plants were let to grow for different times after
exposure, and PrPSc was detected in the leaves by PMCA in duplicates for each
time point. The results show that PrPSc was able to bind to leaves and remained
attached to the living plants for at least 49 days after exposure (Figure 4).
Considering that PrPSc signal was detectable normally in the second or third
round of PMCA without obvious trend in relation to time, we conclude that the
relative amount of PrPSc present in leaves did not appear to change
substantially over time. These data indicate that PrPSc can be retained in
living plants for at least several weeks after a simple contact with prion
contaminated materials, and PrPSc remains competent to drive prion
replication.
Plants Uptake Prions from Contaminated Soil
The experiments described above were done by exposure of the surface of
leaves and roots with different solutions containing prions. To evaluate whether
living plants can uptake PrPSc from contaminated soil, we grew barley grass
plants on soil that was contaminated by addition of 263K brain homogenate.
Plants were grown for 1 or 3 weeks under conditions that carefully prevented any
direct contact of the aerial part of the plant with the soil. After this time,
pieces of stem and leaves were collected and analyzed for the presence of PrPSc
by PMCA. As shown in Figure 5A, all plants grown for 3 weeks in contaminated
soil contained PrPSc in their stem, albeit in small quantities that required
four serial rounds of PMCA for detection. One of the four plants analyzed
contained a detectable amount of PrPSc in the leaves (Figure 5B), indicating
that prions were uptaken from the soil and transported into the aerial parts of
the plants, far from the soil. These results differ from a recent article
reporting that infectious prions were not detectable in above the ground tissues
of wheat plants exposed to CWD prions (Rasmussen et al., 2014). The lack of
detection in this article is most likely due to the low sensitive techniques
(western blots or ELISA) employed to analyze the presence of PrPSc. Indeed, as
we reported previously, PMCA has a power of detection, which is several millions
times higher than western blots or ELISA (Saa´ et al., 2006). In order to
estimate the amount of PrPSc present in stem and leaves coming from contaminated
soil, we performed a quantitative PMCA study, as previously described (Chen et
al., 2010). Unfortunately, by comparing the PMCA amplification in the absence or
the presence of plant tissue, it is possible to conclude that stems and leaves
substantially interfered with the PMCA procedure, and thus the calculation
cannot be very precise (Figure S4). Indeed, after two rounds of PMCA we cannot
detect any protease-resistant PrPSc, but on the third round we observed the
maximum amplification (10 9), presumably because at this round the concentration
of PMCA inhibitors has been reduced enough to permit good amplification. At this
point, we can estimate that the amount of PrPSc that reaches the stem and leaves
from contaminated soil is equivalent to the PrPSc concentration present in a
10 6 to 10 9 dilution of sick brain homogenate. Nevertheless, this result is
interesting, because it indicates that the amount of prions uptaken from soil
and transported to aerial parts of the plant is within the infectious range.
Indeed, titration studies showed that the last infectious dilution of a 263K
brain homogenate is 10 9 (Gregori et al., 2006).
DISCUSSION
This study shows that plants can efficiently bind prions contained in brain
extracts from diverse prion infected animals, including CWD-affected cervids.
PrPSc attached to leaves and roots from wheat grass plants remains capable of
seeding prion replication in vitro. Surprisingly, the small quantity of PrPSc
naturally excreted in urine and feces from sick hamster or cervids was enough to
efficiently contaminate plant tissue. Indeed, our results suggest that the
majority of excreted PrPSc is efficiently captured by plants’ leaves and roots.
Moreover, leaves can be contaminated by spraying them with a prion-containing
extract, and PrPSc remains detectable in living plants for as long as the study
wasperformed (several weeks). Remarkably, prion contaminated plants transmit
prion disease to animals upon ingestion, producing a 100% attack rate and
incubation periods not substantially longer than direct oral administration of
sick brain homogenates. Finally, an unexpected but exciting result was that
plants were able to uptake prions from contaminated soil and transport them to
aerial parts of the plant tissue. Although it may seem farfetched that plants
can uptake proteins from the soil and transport it to the parts above the
ground, there are already published reports of this phenomenon (McLaren et al.,
1960; Jensen and McLaren, 1960; Paungfoo-Lonhienne et al., 2008). The high
resistance of prions to degradation and their ability to efficiently cross
biological barriers may play a role in this process. The mechanism by which
plants bind, retain, uptake, and transport prions is unknown. Weare currently
studying the way in which prions interact with plants using purified,
radioactively labeled PrPSc to determine specificity of the interaction,
association constant, reversibility, saturation, movement, etc.
Epidemiological studies have shown numerous instances of scrapie or CWD
recurrence upon reintroduction of animals on pastures previously exposed to
prion-infected animals. Indeed, reappearance of scrapie has been documented
following fallow periods of up to 16 years (Georgsson et al., 2006), and
pastures were shown to retain infectious CWD prions for at least 2 years after
exposure (Miller et al., 2004). It is likely that the environmentally mediated
transmission of prion diseases depends upon the interaction of prions with
diverse elements, including soil, water, environmental surfaces, various
invertebrate animals, and plants. However, since plants are such an important
component of the environment and also a major source of food for many animal
species, including humans, our results may have far-reaching implications for
animal and human health. Currently, the perception of the risk for
animal-to-humanprion transmissionhas beenmostly limited to consumption or
exposure to contaminated meat; our results indicate that plants might also be an
important vector of transmission that needs to be considered in risk
assessment.
snip...see full text here ;
Grass Plants Bind, Retain, Uptake, and Transport Infectious Prions
Friday, September 27, 2013
Uptake of Prions into Plants
Friday, May 15, 2015 Grass Plants Bind, Retain, Uptake, and Transport
Infectious Prions
Report
*** COMMERCIAL IN CONFIDENCE ***
SPREADING OF UNPROCESSED BLOOD ON LAND
SCRAPIE SEMEN COVER-UP
Immediate and ongoing detection of prions in the blood of hamsters and deer
following oral, nasal, or blood inoculations
Alan M. Elder1, Davin M. Henderson1, Amy V. Nalls1, Edward A. Hoover1,
Anthony E. Kincaid2,3, Jason C. Bartz2 and Candace K. Mathiason1#
+ Author Affiliations 1Department of Microbiology, Immunology and
Pathology, Colorado State University, Fort Collins, CO, 80523, United States of
America 2Medical Microbiology and Immunology, Creighton University, Omaha, NE,
68178, United States of America 3Department of Pharmacy Sciences, Creighton
University, Omaha, NE, 68178, United States of America
ABSTRACT
Infectious prions traverse epithelial barriers to gain access to the
circulatory system, yet the temporal parameters of transepithelial transport and
persistence in the blood over time remains unknown. We used wbRT-QuIC to analyze
whole blood collected from TSE-inoculated deer and hamsters throughout the
entire incubation period for the presence of PrPC-conversion competent amyloid
(PrPC-CCA). We observed PrPC-CCA in the blood of TSE-inoculated hosts throughout
disease course from minutes post exposure to terminal disease.
FOOTNOTES
↵#To whom any correspondence should be addressed:
candace.mathiason@colostate.edu, 1619 Campus Delivery, Fort Collins, Co
80523-1619, 970 491-3975 Copyright © 2015, American Society for Microbiology.
All Rights Reserved.
Thursday, April 30, 2015
*** Immediate and ongoing detection of prions in the blood of hamsters and
deer following oral, nasal, or blood inoculations ***
2001
4. PROPERTIES OF WASTES RELEVANT TO AGRICULTURAL BENEFIT AND ENVIRONMENTAL
IMPACT
European Commission-Directorate-General for Environment WRc Ref: CO
4953-2/11768-1 July 2001 49 4.2.2 Background Wastes from abattoirs include
blood, gut contents, wash waters and sludge from dissolved air flotation
treatment where this process has been used to separate solids from liquid waste
materials of the abattoir. Some wastes such as hoof parts and bone meal are
recycled in other industries (e.g. fertiliser and glue). Landspreading of
abattoir wastes is probably the best practicable environmental option for
small-scale abattoirs but it is likely to be much less appropriate for modern
large-scale abattoir operations. Landspreading of blood and gut contents from
abattoirs is liable to cause public nuisance due to odours and environmental
concerns. If spread on the soil surface it is unsightly and there is potential
for disease transmission. The material should be dealt with as for untreated
sewage sludge and applied to the land by subsurface soil injection or else
incorporated as soon as possible after spreading on the surface of the arable
land. The land-use restrictions as for untreated sewage sludge should apply. The
rate of application of the waste should be in accordance with crop requirements
for nutrients. 4.2.3 Key Properties Waste blood is produced in large quantities
from abattoirs and has various uses including landspreading. Its high fertiliser
value has been known for a long time, and it is one of the more traditional
materials spread on land. Its nitrogen content is extremely high and its levels
of potassium and phosphorus make it a good source of plant nutrients. Nutrients
are also found to be more available than those found in other organic wastes.
Waste stomach contents consist predominantly of partially digested feed or
vegetable matter. As with the blood waste, stomach contents usually contain high
levels of nitrogen, potassium and phosphorus. These nutrients are generally in
well balanced proportions with an N:P:K ratio of around 5:1:1. Moderately high
ammonium nitrogen content is an added benefit. As with many other food
processing industries, large volumes of wash waters are produced, and the term
is often used to describe a wide range of low solid waste materials. This
category can contain dung and urine from animal holding areas and washings from
distribution vehicles. As for the other abattoir wastes, the wash waters contain
a mixture of nitrogen, potassium and phosphorus but at lower concentrations.
4.2.4 Potential Problems From the data above, it is seen that abattoir wastes
contain high levels of nitrogen, potassium and phosphorus. If applied in excess
to plant requirements, these elements can cause potential water pollution
problems, and may also pose a danger to plant health. These wastes also have a
tendency to have a high BOD which makes the waste readily degradable by soil
micro-organisms ; this can rapidly result in anaerobic soil conditions if over
applied. In general, slaughterhouse wastes are a recognised source of
environmental contamination by Salmonella and other zoonotic pathogens (Wray and
Sojka 1977, Edel et al. 1978) Cryptosporidium may occur in gut contents although
not necessarily in infective form. Veterinary ante-mortem inspection at
slaughterhouses ensures that no animal suffering from European
Commission-Directorate-General for Environment WRc Ref: CO 4953-2/11768-1 July
2001 50 notifiable disease or any other disease likely to affect the fitness of
meat is slaughtered for human consumption. However, slaughtered animals may be
symptomless carriers of pathogenic bacteria and therefore slaughterhouse wastes
should be used with caution and with restrictions on land for rearing livestock
or grazing after application. Strict statutory procedures are now enforced at
abattoirs and renderers with the intention of removing, for separate disposal,
components of cattle carcasses which might contain BSE.
2001
4. PROPERTIES OF WASTES RELEVANT TO AGRICULTURAL BENEFIT AND ENVIRONMENTAL
IMPACT
European Commission-Directorate-General for Environment WRc Ref: CO
4953-2/11768-1 July 2001 49 4.2.2 Background Wastes from abattoirs include
blood, gut contents, wash waters and sludge from dissolved air flotation
treatment where this process has been used to separate solids from liquid waste
materials of the abattoir. Some wastes such as hoof parts and bone meal are
recycled in other industries (e.g. fertiliser and glue). Landspreading of
abattoir wastes is probably the best practicable environmental option for
small-scale abattoirs but it is likely to be much less appropriate for modern
large-scale abattoir operations. Landspreading of blood and gut contents from
abattoirs is liable to cause public nuisance due to odours and environmental
concerns. If spread on the soil surface it is unsightly and there is potential
for disease transmission. The material should be dealt with as for untreated
sewage sludge and applied to the land by subsurface soil injection or else
incorporated as soon as possible after spreading on the surface of the arable
land. The land-use restrictions as for untreated sewage sludge should apply. The
rate of application of the waste should be in accordance with crop requirements
for nutrients. 4.2.3 Key Properties Waste blood is produced in large quantities
from abattoirs and has various uses including landspreading. Its high fertiliser
value has been known for a long time, and it is one of the more traditional
materials spread on land. Its nitrogen content is extremely high and its levels
of potassium and phosphorus make it a good source of plant nutrients. Nutrients
are also found to be more available than those found in other organic wastes.
Waste stomach contents consist predominantly of partially digested feed or
vegetable matter. As with the blood waste, stomach contents usually contain high
levels of nitrogen, potassium and phosphorus. These nutrients are generally in
well balanced proportions with an N:P:K ratio of around 5:1:1. Moderately high
ammonium nitrogen content is an added benefit. As with many other food
processing industries, large volumes of wash waters are produced, and the term
is often used to describe a wide range of low solid waste materials. This
category can contain dung and urine from animal holding areas and washings from
distribution vehicles. As for the other abattoir wastes, the wash waters contain
a mixture of nitrogen, potassium and phosphorus but at lower concentrations.
4.2.4 Potential Problems From the data above, it is seen that abattoir wastes
contain high levels of nitrogen, potassium and phosphorus. If applied in excess
to plant requirements, these elements can cause potential water pollution
problems, and may also pose a danger to plant health. These wastes also have a
tendency to have a high BOD which makes the waste readily degradable by soil
micro-organisms ; this can rapidly result in anaerobic soil conditions if over
applied. In general, slaughterhouse wastes are a recognised source of
environmental contamination by Salmonella and other zoonotic pathogens (Wray and
Sojka 1977, Edel et al. 1978) Cryptosporidium may occur in gut contents although
not necessarily in infective form. Veterinary ante-mortem inspection at
slaughterhouses ensures that no animal suffering from European
Commission-Directorate-General for Environment WRc Ref: CO 4953-2/11768-1 July
2001 50 notifiable disease or any other disease likely to affect the fitness of
meat is slaughtered for human consumption. However, slaughtered animals may be
symptomless carriers of pathogenic bacteria and therefore slaughterhouse wastes
should be used with caution and with restrictions on land for rearing livestock
or grazing after application. Strict statutory procedures are now enforced at
abattoirs and renderers with the intention of removing, for separate disposal,
components of cattle carcasses which might contain BSE.
The plant is designed to slaughter a maximum of 400 cattle and 2,000 sheep
per day equivalent to 4,000 units.
Cattle and sheep are delivered to lairage from where they are taken and
slaughtered. Blood is allowed to drain from the slaughtered animals into an
underground collection trough. Blood (process) from here is vacuum pumped to the
blood treatment plant where anti-coagulant is added after which it is chilled
and stored on-site in an overground collection tank. Processed blood is
subsequently removed from the site and transported by sealed tanker to APC
Technologies Craigavon, Co. Armagh, for further processing.
Blood (waste) arising primarily from washing operations is directed to an
overground collection tank. From here it is transported to Ronan Industries,
Cashel, Co. Tipperary for further processing for use in animal feed.
This office has on file a notarized affidavit from [manufacturer/exporter]
verifying the accuracy of the statements below.
1. The certified products are U.S. origin and are [insert type of blood
product, e.g., whole blood; serum; plasma; such as
albumin or globulin].
2. The products are intended to be used for [insert intended end use, e.g.,
in-vitro purposes (laboratory/diagnostics); animal feed].
3. The products were manufactured from bovine blood or one of its
components or derivatives.
4. The blood used in the manufacture of the products was derived from
healthy cattle subjected to slaughter inspection in an official establishment
under the control of the competent authority.
5. The animals from which the blood was derived were not subjected to a
stunning process, prior to slaughter, with a device injecting air or gas into
the cranial cavity, or to a pithing process.
6. The products were manufactured in a facility approved by the competent
authority and meet the requirements of the United States for domestic sale and
use.
7. The products were manufactured in accordance with U.S. laws and
regulations intended to ensure that they are unlikely to transmit disease
agents, including transmissible spongiform encephalopathies (TSEs).
8. Adequate precautions were taken following processing to prevent product
contamination with microbiological pathogens.
LOL
The total volume of feeds produced by the 83 feed manufacturers in 1990 was
about 6.4 million tons. Complete feeds made up 56 percent of the total volume,
and supplements/concentrates made up another 38 percent. Approximately 80
percent of the feed manufactures sold their products within 150 miles of the
production facility, and 85 percent acquired their meat and bone meal/meat
by-products (MBM/MBP) from within 150 miles.
Almost all of the manufacturers used MBM/MBP in their feeds. Although most
(63%) of the MBM/MBP went into swine feeds, 15 percent went into beef cattle
feeds and 12 percent into dairy cattle feeds. About three-fourths of the
manufacturers did not require any animal species to be excluded from the MBM/MBP
that was used in the feeds. Of the feed products intended for dairy cattle, 53
percent included MBM/MBP from unknown species that did not exclude sheep.
Fifty-six (56) percent of beef cattle feeds incorporated MBM/MBP from unknown
species which did not exclude sheep.
LOL
Blood, not used in the manufacture of feed-stuff, is not an animal
by-product if the blood is from healthy animals and may be eligible to be
landspread under the exemption of WMLR 1994. If blood is from diseased animals,
it is an animal by-product.
SNIP...
Strict procedures are now enforced at abattoirs and renderers with the
intention of removing, for separate disposal, components of cattle carcasses
which might contain BSE prions. The term Specified Bovine Material (SBM), now
called Specified Risk Material (SRM), is used to refer to these parts of the
carcass. An example of these statutory procedures is The Specified Bovine
Material (No. 2) Order 1996 (SI 1996 No. 1192) which came into effect on 1 May
1996. Further legislation (Article 2e SBM (No. 3) Order 1996) includes the need
to ensure that European Commission - Directorate-General for Environment WRc
Ref:CO4953-2/11768-1 July 2001 508 trapped abattoir waste (i.e. caught in
screens and drain traps in areas handling SBM) is dealt with separately as SBM
and is not discharged onto land.
LOL
SCIENTIFIC REPORT OF EFSA
Protocol for further laboratory investigations into the distribution of
infectivity of Atypical BSE1
European Food Safety Authority2,3
European Food Safety Authority (EFSA), Parma, Italy
Suggested citation: European Food Safety Authority, 2014. Protocol for
further laboratory investigations into the distribution
of infectivity of Atypical BSE. EFSA Journal 2014;12(7):3798, 55 pp.
doi:10.2903/j.efsa.2014.3798 Available online:
www.efsa.europa.eu/efsajournal
© European Food Safety Authority, 2014
SCIENTIFIC REPORT OF EFSA
Protocol for further laboratory investigations into the distribution of
infectivity of Atypical BSE1
European Food Safety Authority2,3
European Food Safety Authority (EFSA), Parma, Italy
ABSTRACT
Information on the pathogenesis and tissue distribution of Atypical Bovine
Spongiform Encephalopathy (BSE) in cattle through the study of field cases and
experimental transmission studies is lacking. The latter are limited to
transmission of Atypical BSE through intracerebral (i.c.) inoculation of cattle.
All data currently available relate to the presence or absence of PrPSc, but do
not quantify relative amounts of PrPSc or levels of infectivity. A laboratory
protocol for further studies is recommended, to allow the assessment of the
relative infectious titre, PrPSc accumulation and prion seeding activity in the
tissues of cattle that developed H-BSE or L-BSE (using posterior brainstem as a
reference). Tissues to be covered by those studies are categorised in three
priorities, based on their inclusion in the list of specific risk material in
cattle, on the presence of infectivity, or PrPSc presence, demonstrated in
Atypical BSEs or other Transmissible Spongiform Encephalopathies (TSEs) in
ruminants, and on the importance in terms of input into the food chain in the
EU. The protocol provides details in terms of the minimum number of animals to
be tested, processing and preparation of tissues, and methods to be used to
identify abnormal PrP and quantify infectivity, also depending on the expected
level of infectivity and amount of tissue available for analysis. It is
recommended that, through the implementation of the protocol, information should
also be obtained on the performance of currently validated rapid tests for TSE
active surveillance in cattle/bioassay for detecting H-BSE and L-BSE
agents.
© European Food Safety Authority, 2014
KEY WORDS
Atypical BSE, cattle, H-BSE, L-BSE, laboratory protocol, prion
Thursday, April 30, 2015
*** Immediate and ongoing detection of prions in the blood of hamsters and
deer following oral, nasal, or blood inoculations
Thursday, September 18, 2014
Development of Dose-Response Models of Creutzfeldt-Jakob Disease Infection
in Nonhuman Primates for Assessing the Risk of Transfusion-Transmitted variant
Creutzfeldt-Jakob Disease
Development of Dose-Response Models of Creutzfeldt-Jakob Disease Infection
in Nonhuman Primates for Assessing the Risk of Transfusion-Transmitted variant
Creutzfeldt-Jakob Disease
Yin Huanga, Luisa Gregorib, Steven A. Andersona, David M. Asherb and Hong
Yanga# + Author Affiliations
Office of Biostatistics & Epidemiology, U.S. Food and Drug
Administration, Silver Spring, Maryland, USAa Office of Blood Research and
Review, U.S. Food and Drug Administration, Silver Spring, Maryland, USAb
ABSTRACT Estimates for the risk of transmitting variant Creutzfeldt-Jakob
disease (vCJD) via blood transfusion have largely relied on data from rodent
experiments, but the relationship between dose (amount of infected blood) and
response (vCJD infection) has never been well quantified. The goal of this study
was to develop a dose-response model based on nonhuman primate data to better
estimate the likelihood of transfusion-transmitted vCJD (TTvCJD) in humans. Our
model used dose-response data from nonhuman primates inoculated intracerebrally
(IC) with brain tissues of patients with sporadic and familial CJD. We analyzed
the data statistically using a beta-Poisson dose-response model. We further
adjusted model parameters to account for the differences in infectivity between
blood and brain tissue and in transmission efficiency between intravenous (IV)
and IC routes to estimate dose-dependent TTvCJD infection. *** The model
estimates a mean infection rate of 76% among recipients who receive one unit of
whole blood collected from an infected donor near the end of the incubation
period. The nonhuman primate model provides estimates that are more consistent
with those derived from a risk analysis of transfused non-leukoreduced red blood
cells in United Kingdom compared to prior estimates based on rodent models.
IMPORTANCE TTvCJD was recently identified as one of three emerging
infectious diseases posing the greatest immediate threat to the safety of the
blood supply. Cases of TTvCJD were reported in recipients of non-leukoreduced
red blood cells and coagulation Factor VIII manufactured from blood of UK
donors. As the quantity of abnormal prions (the causative agent of TTvCJD)
varies significantly in different blood components and products, it is necessary
to quantify the dose-response relationship for a wide range of doses for the
vCJD agent in transfused blood and plasma derivatives. In this paper we suggest
the first mechanistic dose-response model for TTvCJD infection based on data
from experiments with nonhuman primates. This new model may improve estimates of
the possible risk to humans.
FOOTNOTES ↵#Address correspondence to Hong Yang, Hong.Yang@fda.hhs.gov
Copyright © 2014, American Society for Microbiology. All Rights Reserved.
can anyone say TSEAC meeting in 2014 ???
Tuesday, April 21, 2015 Transmissible Spongiform Encephalopathy Advisory
Committee TSEAC MEETING SCHEDULED FOR June 1, 2015
Tuesday, December 30, 2014
TSEAC USA Reason For Recalls Blood products, collected from a donors
considered to be at increased risk for Creutzfeldt-Jakob Disease (CJD), were
distributed END OF YEAR REPORT 2014
Sunday, March 09, 2014
A Creutzfeldt-Jakob Disease (CJD) Lookback Study: Assessing the Risk of
Blood Borne Transmission of Classic Forms of Creutzfeldt-Jakob Disease
FDA TSEAC CIRCUS AND TRAVELING ROAD SHOW FOR THE TSE PRION DISEASES
Sunday, June 9, 2013
TSEAC March 14, 2013: Transmissible Spongiform Encephalopathies Advisory
Committee Meeting Webcast
see more history TSEAC here ;
Saturday, May 09, 2015
*** Psychiatric Symptoms in Patients With Sporadic Creutzfeldt-Jakob
Disease ***
PRION2015 FORT COLLINS
Wednesday May 27
14:45 Jean-Phillipe Deslys Atomic Energy Commission, France,
Transmission of prions to primates after extended silent incubation
periods: * IMPLICATIONS FOR BSE AND SCRAPIE RISK ASSESSMENT IN HUMAN
POPULATIONS.
16:45
Quingzhong Kong Case Western Reserve University
Zoonotic Potential of CWD Prions
*** Kuru Video ***
Kuru: The Science and The Sorcery
*** Scrapie Video
*** Human Mad Cow Video
*** USA sporadic CJD MAD COW DISEASE HAS HUGE PROBLEM Video
2014
***Moreover, L-BSE has been transmitted more easily to transgenic mice
overexpressing a human PrP [13,14] or to primates [15,16] than C-BSE.
***It has been suggested that some sporadic CJD subtypes in humans may
result from an exposure to the L-BSE agent.
*** Lending support to this hypothesis, pathological and biochemical
similarities have been observed between L-BSE and an sCJD subtype (MV genotype
at codon 129 of PRNP) [17], and between L-BSE infected non-human primate and
another sCJD subtype (MM genotype) [15].
snip...
Monday, October 10, 2011
EFSA Journal 2011 The European Response to BSE: A Success Story
snip...
EFSA and the European Centre for Disease Prevention and Control (ECDC)
recently delivered a scientific opinion on any possible epidemiological or
molecular association between TSEs in animals and humans (EFSA Panel on
Biological Hazards (BIOHAZ) and ECDC, 2011). This opinion confirmed Classical
BSE prions as the only TSE agents demonstrated to be zoonotic so far
*** but the possibility that a small proportion of human cases so far
classified as "sporadic" CJD are of zoonotic origin could not be excluded.
*** Moreover, transmission experiments to non-human primates suggest that
some TSE agents in addition to Classical BSE prions in cattle (namely L-type
Atypical BSE, Classical BSE in sheep, transmissible mink encephalopathy (TME)
and chronic wasting disease (CWD) agents) might have zoonotic potential.
snip...
Thursday, August 12, 2010
Seven main threats for the future linked to prions
First threat
The TSE road map defining the evolution of European policy for protection
against prion diseases is based on a certain numbers of hypotheses some of which
may turn out to be erroneous. In particular, a form of BSE (called atypical
Bovine Spongiform Encephalopathy), recently identified by systematic testing in
aged cattle without clinical signs, may be the origin of classical BSE and thus
potentially constitute a reservoir, which may be impossible to eradicate if a
sporadic origin is confirmed.
*** Also, a link is suspected between atypical BSE and some apparently
sporadic cases of Creutzfeldt-Jakob disease in humans.
*** These atypical BSE cases constitute an unforeseen first threat that
could sharply modify the European approach to prion diseases.
Second threat
snip...
*** Creutzfeldt-Jakob Disease Public Health Crisis VIDEO
Tuesday, December 16, 2014
Evidence for zoonotic potential of ovine scrapie prions
Hervé Cassard,1, n1 Juan-Maria Torres,2, n1 Caroline Lacroux,1, Jean-Yves
Douet,1, Sylvie L. Benestad,3, Frédéric Lantier,4, Séverine Lugan,1, Isabelle
Lantier,4, Pierrette Costes,1, Naima Aron,1, Fabienne Reine,5, Laetitia
Herzog,5, Juan-Carlos Espinosa,2, Vincent Beringue5, & Olivier
Andréoletti1, Affiliations Contributions Corresponding author Journal name:
Nature Communications Volume: 5, Article number: 5821 DOI:
doi:10.1038/ncomms6821 Received 07 August 2014 Accepted 10 November 2014
Published 16 December 2014 Article tools Citation Reprints Rights &
permissions Article metrics
Abstract
Although Bovine Spongiform Encephalopathy (BSE) is the cause of variant
Creutzfeldt Jakob disease (vCJD) in humans, the zoonotic potential of scrapie
prions remains unknown. Mice genetically engineered to overexpress the human
prion protein (tgHu) have emerged as highly relevant models for gauging the
capacity of prions to transmit to humans. These models can propagate human
prions without any apparent transmission barrier and have been used used to
confirm the zoonotic ability of BSE. Here we show that a panel of sheep scrapie
prions transmit to several tgHu mice models with an efficiency comparable to
that of cattle BSE. The serial transmission of different scrapie isolates in
these mice led to the propagation of prions that are phenotypically identical to
those causing sporadic CJD (sCJD) in humans. These results demonstrate that
scrapie prions have a zoonotic potential and raise new questions about the
possible link between animal and human prions.
Subject terms: Biological sciences• Medical research At a glance
why do we not want to do TSE transmission studies on chimpanzees $
5. A positive result from a chimpanzee challenged severly would likely
create alarm in some circles even if the result could not be interpreted for
man. I have a view that all these agents could be transmitted provided a large
enough dose by appropriate routes was given and the animals kept long enough.
Until the mechanisms of the species barrier are more clearly understood it might
be best to retain that hypothesis.
snip...
R. BRADLEY
Suspect symptoms
What if you can catch old-fashioned CJD by eating meat from a sheep
infected with scrapie?
28 Mar 01 Most doctors believe that sCJD is caused by a prion protein
deforming by chance into a killer. But Singeltary thinks otherwise. He is one of
a number of campaigners who say that some sCJD, like the variant CJD related to
BSE, is caused by eating meat from infected animals. Their suspicions have
focused on sheep carrying scrapie, a BSE-like disease that is widespread in
flocks across Europe and North America.
Now scientists in France have stumbled across new evidence that adds weight
to the campaigners' fears. To their complete surprise, the researchers found
that one strain of scrapie causes the same brain damage in mice as sCJD.
"This means we cannot rule out that at least some sCJD may be caused by
some strains of scrapie," says team member Jean-Philippe Deslys of the French
Atomic Energy Commission's medical research laboratory in Fontenay-aux-Roses,
south-west of Paris. Hans Kretschmar of the University of Göttingen, who
coordinates CJD surveillance in Germany, is so concerned by the findings that he
now wants to trawl back through past sCJD cases to see if any might have been
caused by eating infected mutton or lamb...
2001
Suspect symptoms
What if you can catch old-fashioned CJD by eating meat from a sheep
infected with scrapie?
28 Mar 01
Like lambs to the slaughter
31 March 2001
by Debora MacKenzie Magazine issue 2284.
FOUR years ago, Terry Singeltary watched his mother die horribly from a
degenerative brain disease. Doctors told him it was Alzheimer's, but Singeltary
was suspicious. The diagnosis didn't fit her violent symptoms, and he demanded
an autopsy. It showed she had died of sporadic Creutzfeldt-Jakob disease.
Most doctors believe that sCJD is caused by a prion protein deforming by
chance into a killer. But Singeltary thinks otherwise. He is one of a number of
campaigners who say that some sCJD, like the variant CJD related to BSE, is
caused by eating meat from infected animals. Their suspicions have focused on
sheep carrying scrapie, a BSE-like disease that is widespread in flocks across
Europe and North America.
Now scientists in France have stumbled across new evidence that adds weight
to the campaigners' fears. To their complete surprise, the researchers found
that one strain of scrapie causes the same brain damage in mice as sCJD.
"This means we cannot rule out that at least some sCJD may be caused by
some strains of scrapie," says team member Jean-Philippe Deslys of the French
Atomic Energy Commission's medical research laboratory in Fontenay-aux-Roses,
south-west of Paris. Hans Kretschmar of the University of Göttingen, who
coordinates CJD surveillance in Germany, is so concerned by the findings that he
now wants to trawl back through past sCJD cases to see if any might have been
caused by eating infected mutton or lamb.
Scrapie has been around for centuries and until now there has been no
evidence that it poses a risk to human health. But if the French finding means
that scrapie can cause sCJD in people, countries around the world may have
overlooked a CJD crisis to rival that caused by BSE.
Deslys and colleagues were originally studying vCJD, not sCJD. They
injected the brains of macaque monkeys with brain from BSE cattle, and from
French and British vCJD patients. The brain damage and clinical symptoms in the
monkeys were the same for all three. Mice injected with the original sets of
brain tissue or with infected monkey brain also developed the same symptoms.
As a control experiment, the team also injected mice with brain tissue from
people and animals with other prion diseases: a French case of sCJD; a French
patient who caught sCJD from human-derived growth hormone; sheep with a French
strain of scrapie; and mice carrying a prion derived from an American scrapie
strain. As expected, they all affected the brain in a different way from BSE and
vCJD. But while the American strain of scrapie caused different damage from
sCJD, the French strain produced exactly the same pathology.
"The main evidence that scrapie does not affect humans has been
epidemiology," says Moira Bruce of the neuropathogenesis unit of the Institute
for Animal Health in Edinburgh, who was a member of the same team as Deslys.
"You see about the same incidence of the disease everywhere, whether or not
there are many sheep, and in countries such as New Zealand with no scrapie." In
the only previous comparisons of sCJD and scrapie in mice, Bruce found they were
dissimilar.
But there are more than 20 strains of scrapie, and six of sCJD. "You would
not necessarily see a relationship between the two with epidemiology if only
some strains affect only some people," says Deslys. Bruce is cautious about the
mouse results, but agrees they require further investigation. Other trials of
scrapie and sCJD in mice, she says, are in progress.
People can have three different genetic variations of the human prion
protein, and each type of protein can fold up two different ways. Kretschmar has
found that these six combinations correspond to six clinical types of sCJD: each
type of normal prion produces a particular pathology when it spontaneously
deforms to produce sCJD.
But if these proteins deform because of infection with a disease-causing
prion, the relationship between pathology and prion type should be different, as
it is in vCJD. "If we look at brain samples from sporadic CJD cases and find
some that do not fit the pattern," says Kretschmar, "that could mean they were
caused by infection."
There are 250 deaths per year from sCJD in the US, and a similar incidence
elsewhere. Singeltary and other US activists think that some of these people
died after eating contaminated meat or "nutritional" pills containing dried
animal brain. Governments will have a hard time facing activists like Singeltary
if it turns out that some sCJD isn't as spontaneous as doctors have insisted.
Deslys's work on macaques also provides further proof that the human
disease vCJD is caused by BSE. And the experiments showed that vCJD is much more
virulent to primates than BSE, even when injected into the bloodstream rather
than the brain. This, says Deslys, means that there is an even bigger risk than
we thought that vCJD can be passed from one patient to another through
contaminated blood transfusions and surgical instruments.
Thursday, March 20, 2014
CHRONIC WASTING DISEASE CWD TSE PRION OF CERVID AND THE POTENTIAL FOR HUMAN
TRANSMISSION THEREFROM 2014
Tuesday, July 01, 2014
*** CHRONIC WASTING DISEASE CWD TSE PRION DISEASE, GAME FARMS, AND
POTENTIAL RISK FACTORS THERE FROM ***
Thursday, July 03, 2014
*** How Chronic Wasting Disease is affecting deer population and what’s the
risk to humans and pets? ***
Thursday
CWD TO HUMANS, AND RISK FACTORS THERE FROM (see latest science)
Tuesday, November 04, 2014
*** Six-year follow-up of a point-source exposure to CWD contaminated
venison in an Upstate New York community: risk behaviours and health outcomes
2005–2011
Thursday, April 30, 2015
Immediate and ongoing detection of prions in the blood of hamsters and deer
following oral, nasal, or blood inoculations
Friday, January 30, 2015
*** Scrapie: a particularly persistent pathogen ***
Sunday, April 12, 2015
*** Research Project: Transmission, Differentiation, and Pathobiology of
Transmissible Spongiform Encephalopathies 2014 Annual Report ***
http://transmissiblespongiformencephalopathy.blogspot.com/2015/04/research-project-transmission.html
Saturday, April 11, 2015
*** ISU veterinary researchers study retinal scans as early detection
method for mad cow disease
Sunday, November 23, 2014
*** Confirmed Variant Creutzfeldt-Jakob Disease (variant CJD) Case in Texas
in June 2014 confirmed as USA case NOT European ***
Monday, November 3, 2014
USA CJD TSE PRION UNIT, TEXAS, SURVEILLANCE UPDATE NOVEMBER 2014
National Prion Disease Pathology Surveillance Center Cases Examined1
(October 7, 2014)
***6 Includes 11 cases in which the diagnosis is pending, and 19
inconclusive cases;
***7 Includes 12 (11 from 2014) cases with type determination pending in
which the diagnosis of vCJD has been excluded.
***The sporadic cases include 2660 cases of sporadic Creutzfeldt-Jakob
disease (sCJD),
***50 cases of Variably Protease-Sensitive Prionopathy (VPSPr)
***and 21 cases of sporadic Fatal Insomnia (sFI).
Thursday, January 15, 2015
41-year-old Navy Commander with sporadic Creutzfeldt–Jakob disease CJD TSE
Prion: Case Report
Subject: *** Becky Lockhart 46, Utah’s first female House speaker, dies
diagnosed with the extremely rare Creutzfeldt-Jakob disease aka mad cow type
disease
what is CJD ? just ask USDA inc., and the OIE, they are still feeding the
public and the media industry fed junk science that is 30 years old.
why doesn’t some of you try reading the facts, instead of rubber stamping
everything the USDA inc says.
sporadic CJD has now been linked to BSE aka mad cow disease, Scrapie, and
there is much concern now for CWD and risk factor for humans.
My sincere condolences to the family and friends of the House Speaker Becky
Lockhart. I am deeply saddened hear this.
with that said, with great respect, I must ask each and every one of you
Politicians that are so deeply saddened to hear of this needless death of the
Honorable House Speaker Becky Lockhart, really, cry me a friggen river. I am
seriously going to ask you all this...I have been diplomatic for about 17 years
and it has got no where. people are still dying. so, are you all stupid or
what??? how many more need to die ??? how much is global trade of beef and other
meat products that are not tested for the TSE prion disease, how much and how
many bodies is this market worth?
Saturday, January 17, 2015
*** Becky Lockhart 46, Utah’s first female House speaker, dies diagnosed
with the extremely rare Creutzfeldt-Jakob disease
*** ALERT new variant Creutzfeldt Jakob Disease nvCJD or vCJD, sporadic CJD
strains, TSE prion aka Mad Cow Disease United States of America Update December
14, 2014 Report ***
Tuesday, November 04, 2014
Towards an Age-Dependent Transmission Model of Acquired and Sporadic
Creutzfeldt-Jakob Disease
Thursday, January 22, 2015
Transmission properties of atypical Creutzfeldt-Jakob disease: a clue to
disease etiology?
Sunday, July 06, 2014
Dietary Risk Factors for Sporadic Creutzfeldt-Jakob Disease: A Confirmatory
Case-Control Study
Conclusions—The a priori hypotheses were supported.
*Consumption of various meat products may be one method of transmission of
the infectious agent for sCJD.
PLEASE REMEMBER ;
The Akron, Ohio-based CJD Foundation said the Center for Disease Control
revised that number in October of 2004 to about one in 9,000 CJD cases per year
in the population group age 55 and older.
HAVE YOU GOT YOUR CJD QUESTIONNAIRE ASKING REAL QUESTIONS PERTAINING TO
ROUTE AND SOURCE OF THE TSE AGENT THAT KILLED YOUR LOVED ONE ???
if not, why not...
Friday, November 30, 2007
CJD QUESTIONNAIRE USA CWRU AND CJD FOUNDATION
Friday, January 10, 2014
vpspr, sgss, sffi, TSE, an iatrogenic by-product of gss, ffi, familial type
prion disease, what it ???
Self-Propagative Replication of Ab Oligomers Suggests Potential
Transmissibility in Alzheimer Disease
Received July 24, 2014; Accepted September 16, 2014; Published November 3,
2014
Singeltary comment ;
Saturday, December 13, 2014
Terry S. Singeltary Sr. Publications TSE prion disease
Diagnosis and Reporting of Creutzfeldt-Jakob Disease
Singeltary, Sr et al. JAMA.2001; 285: 733-734. Vol. 285 No. 6, February 14,
2001 JAMA
snip...
Tuesday, April 14, 2015
Transmissible Spongiform Encephalopthy TSE Prion Disease
Wednesday, April 15, 2015
KURU Transmissible Spongiform Encephalopthy TSE Prion Disease
Sunday, May 3, 2015
PRION2015 FORT COLLINS
Saturday, December 13, 2014
Terry S. Singeltary Sr. Publications TSE prion disease
Diagnosis and Reporting of Creutzfeldt-Jakob Disease
Singeltary, Sr et al. JAMA.2001; 285: 733-734. Vol. 285 No. 6, February 14,
2001 JAMA
snip...
and that’s why they call it sporadic creutzfeldt-jakob disease sCJD.
simply meaning unknown route and source of the TSE prion agent. ...
TSS
posted by Terry S. Singeltary Sr. at
8:30 AM
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