Texas CWD Captive, and then there were 4 ?
Texas CWD Captive, and then there were 4 ?
Rancher: 4 deer from Medina County test positive for CWD
Chronic wasting disease outbreak being monitored by Texas Animal Health
Commission, Texas Parks and Wildlife Department
Updated TODAY, 2:58 PM
By Pilar Arias
Reporter
SAN ANTONIO - Texas Mountain Ranch owner Robert Patterson says four deer
from his facility in Medina County have tested positive for chronic wasting
disease.
Patterson has been working with the Texas Animal Health Commission and
Texas Parks and Wildlife Department since one of his captive white-tailed deer
tested positive for CWD in June.
snip...
He said 42 deer have been killed and tested since July 28, and three
additional positives were the result. He added that all four deer confirmed to
have the disease were males from the same father, which leads him to believe the
problem is genetic.
snip...
why in the world was there not already a CWD EMERGENCY RESPONSE PLAN FOR
CAPTIVES already set up and ready to go in Texas ?
what’s the cwd task force been doing?
what happened behind closed door sessions?
why is it taking so long for the TAHC et al to report these results ? (can
it be we are waiting to find someone that can find a false negative, maybe like
the mad cow in Texas that was covered up for 7 months so the BSE MRR policy
could be validated first? I’m just guessing out loud) of the fact that Texas
waited 10 years to finally test for CWD, right where I told them a decade
earlier where it would be. for 10 years that cwd walked or was trucked across
Texas. or longer.
when is the testing on the rest of the herd be done?
what about cohorts of cwd positives, their trace outs, those herds?
why aren’t those herds quarantined yet ?
where are the reports?
what’s the rush, right $$$
let’s see just how much, and how far we can spread this CWD TSE
prion.
this is absolutely crazy.
Thursday, August 06, 2015
WE HAVE LOST TEXAS TO CWD TASK FORCE CATERING TO INDUSTRY
Friday, August 7, 2015
Transgenic Mouse Bioassay: Evidence That Rabbits Are Susceptible to a
Variety of Prion Isolates
PrPSc detection and infectivity in semen from scrapie-infected sheep
Richard Rubenstein,1 Marie S. Bulgin,2 Binggong Chang,1 Sharon
Sorensen-Melson,2 Robert B. Petersen3 and Giuseppe LaFauci4 Correspondence
Richard Rubenstein richard.rubenstein@downstate. edu Received 13 October 2011
Accepted 3 February 2012 1Departments of Neurology and Physiology/Pharmacology,
SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, USA
2University of Idaho, Caine Veterinary Teaching and Research Center, 1020 E.
Homedale Road, Caldwell, ID 83607, USA 3Departments of Pathology Neuroscience,
and Neurology, Case Western Reserve University, 2103 Cornell Road, Cleveland, OH
44120, USA 4New York State Institute for Basic Research in Developmental
Disabilities, 1050 Forest Hill Road, Staten Island, NY 10314, USA
A scrapie-positive ewe was found in a flock that had been scrapie-free for
13 years, but housed adjacent to scrapie-positive animals, separated by a wire
fence. Live animal testing of the entire flock of 24 animals revealed seven more
subclinical scrapie-positive ewes. We hypothesized that they may have contracted
the disease from scrapie-positive rams used for breeding 4 months prior,
possibly through the semen. The genotypes of the ewe flock were highly
scrapiesusceptible and the rams were infected with the ‘Caine’ scrapie strain
having a short incubation time of 4.3–14.6 months in sheep with 136/171 VQ/VQ
and AQ/VQ genotypes. PrPSc accumulates in a variety of tissues in addition to
the central nervous system. Although transmission of prion diseases, or
transmissible spongiform encephalopathies, has been achieved via peripheral
organ or tissue homogenates as well as by blood transfusion, neither infectivity
nor PrPSc have been found in semen from scrapie-infected animals. Using serial
protein misfolding cyclic amplification followed by a surround optical fibre
immunoassay, we demonstrate that semen from rams infected with a
short-incubation-time scrapie strain contains prion disease-associatedseeding
activity that generated PrPSc in sPMCA (serial protein misfolding cyclic
amplification). Injection of the ovinized transgenic mouse line TgSShpPrP with
semen from scrapie-infected sheep resulted in PrPSc-seeding activity in clinical
and, probably as a result of the low titre, nonclinical mouse brain. These
results suggest that the transmissible agent, or at least the seeding activity,
for sheep scrapie is present in semen. This may be a strain-specific phenomenon.
RESULTS
The use of SOFIA alone to detect PrPSc in semen samples from
scrapie-infected sheep was unsuccessful (data not shown). In previous studies
(Rubenstein et al., 2010, 2011), we also reported that SOFIA alone was unable to
detect PrPSc in blood and urine from scrapie-infected sheep and cervids with
chronic wasting disease. However, in combination with sPMCA, which utilizes
PASA, PrPSc in blood and urine was detectable. The level of PrPSc detected was
dependent on the number of serial cycles of PMCA. Furthermore, amplification is
efficient independent of the genotypes of the PrPC and PrPSc (Rubenstein et al.,
2010, 2011). The level of PrPSc measured by SOFIA following sPMCA80 and sPMCA120
(Fig. 1) was similar to that found in urine (Rubenstein et al., 2011) and
significantly less than that found in blood (Rubenstein et al., 2010) from
scrapieinfected sheep. This suggests that the PASA found in the semen of these
animals is similar to that for urine from infected animals, approximately 1
attogram ml21. However, it is not possible to determine whether prion-seeding
activity is actually PrPSc (Rubenstein et al., 2011).
The ovinized PrP transgenic mouse line, TgSShpPrP, was analysed for ovine
PrPC expression. Serial twofold dilutions of brain homogenates from TgSShpPrP
mice and wild-type (WT) mice on the same FVB genetic background were analysed by
Western blotting. PrPC was immunostained using the PrP-specific mAb,
08-6/2F7-2F11. The dilutions producing equivalent PrPC immunostaining between
the two mouse lines indicate that the level of expression of sheep PrPC in the
brains of TgSShpPrP mice is approximately fourfold greater than mouse PrPC in
the WT mice (Fig. 2).
The ability of TgSShpPrP mice to support replication of the sheep prion
agent was analysed and is summarized in Table 2. Brain homogenate from
scrapie-infected ram #8001, which was positive for PK-resistant PrPSc by Western
blotting (Fig. 3a, lanes 3–4), was used to infect TgSShpPrP mice. Injection of
nine TgSShpPrP mice by the intracerebral (IC) route with a 10% brain homogenate
from ram #8001 had a mean incubation period of 167±7 days, indicating that these
mice support replication of the ovine prion agent. Western blotting of brain
from clinical TgSShpPrP mice demonstrated the presence of PK-resistant PrPSc
(Fig. 3a, lane 8). In contrast, TgSShpPrP mice showed no clinical signs or PrPSc
after over 520 days following IC injection of either a 10% normal sheep brain
(NSB) homogenate or a 10% ME7 mouse-adapted scrapie strain brain homogenate
(Table 2).
Semen samples from scrapie-positive rams (# 3272, 7031, 7035 and 8001) were
injected IC into TgSShpPrP mice (five mice per semen sample) to bioassay for
prion infectivity (Table 2). One TgSShpPrP mouse from the ram #7031 semen sample
group and one from the ram #3272 semeninjected group showed clinical signs after
an incubation period of 310 and 655 days, respectively. Western blotting of the
brain from these mice demonstrated PK-resistant PrPSc (Fig. 3a and b).
Interestingly, the mouse from the ram #3272 group did not display any clinical
signs at 648 days post-injection (p.i.), but by 655 days p.i. the animal was at
the end stage of clinical disease. This is about half the time it typically
takes mice to proceed through the same clinical disease course. The remaining
mice from these groups, as well as all the mice from the other ram
semen-injected groups did not display any clinical disease throughout the entire
experiment (.670 days p.i.).
Animals were randomly selected from the groups of nonclinical TgSShpPrP
mice injected with semen samples from ram #3272 (385 days p.i.), 7031 (385 days
p.i.), 7035 (350 days p.i.) and 8001 (335 days p.i.). TgSShpPrP mouse brain
homogenates were prepared and analysed by Western blotting for the presence of
PrPSc. PK-resistant PrPSc, indicative of prion disease, was detected in the
brain homogenates of all the semen-injected non-clinical TgSShpPrP mice (Fig.
3b, lane 2; Fig. 4 lanes 6, 8, 10 and 12). Interestingly, all of the
PK-resistant PrPSc profiles from the non-clinical mice injected with different
semen samples were similar with a prominent protein band at 27–30 kDa,
representing the truncated diglycosylated PrP, a less intense immunostained band
at 23–25 kDa indicative of the truncated monoglycosylated form of PrP and a
lighter, truncated unglycosylated PrP form at 19–20 kDa. Additional analysis of
random, non-clinical TgSShpPrP mice selected from the semen-injected groups at
523 days p.i. produced similar positive Western blot results and the same PrPSc
profile (data not shown). In contrast, and as expected, clinical TgSShpPrP mice
injected with either ram #8001 brain homogenate (Fig. 3a, lane 8; Fig. 4, lane
2), or semen from ram #7031 (Fig. 3a, lane 10) and 3272 (Fig. 3b, lane 6) had a
PK-resistant PrPSc pattern consisting of three intensely immunostained bands. A
similar pattern of three intensely immunostained bands was also seen in a
nonclinical mouse from ram #3272 semen-injected group at 648 days p.i. (Fig. 3b,
lane 4), just 7 days before another mouse from this group displayed extensive
clinical disease. None of the five TgSShpPrP mice/group injected IC with semen
from the uninfected rams displayed any clinical signs or PK-resistant PrPSc by
650 days p.i. (data not shown). The IHCs performed on all reproductive tissues
(prostrate, epididymis, vesicular glands and testes) of the above rams were
negative for PrPSc (Table S2). Ram #8031 (ARQ/ ARQ) also tested negative by IHC
on RAMALT and third eyelid and his brain and all other tissues tested were IHC
negative for PrPSc when euthanized following semen collection (Table S2).
Western blot analysis of brain tissue from ram #8031 was negative for
PK-resistant PrPSc. Western blotting of the samples following 40 cycles of
serial PMCA (sPMCA40) using ram #8031 brain tissue as a seed was also negative
for PK-resistant PrPSc (data not shown) presumably due to the limitations of
assay sensitivity. However, using sPMCA40, PASA in ram #8031 brain tissue could
be detected and the product (i.e. PrPSc) measured by SOFIA with signal
intensities (sample/background) of 39.8±2.1 compared with 1.01±0.02 for
uninfected control sheep brain.
DISCUSSION
The decision to use rams from the scrapie-positive flock to breed ewes in
the sentinel flock resulted from the scarcity and difficulty of finding 136V/V
and A/V rams in private certified scrapie-free flocks and the prevalent belief
at that time that rams do not transmit scrapie as indicated by the National
Scrapie Surveillance Plan (2010). Current USDA Scrapie Eradication Uniform
Methods and Rules (2005) do not require the quarantining of flocks following the
introduction of exposed rams. Furthermore, the European Food Safety Authority
recently reached the same conclusion that the risk of TSE transmission
associated with semen and embryos collected from classical scrapie incubating
sheep and goats ranges from negligible to low [EFSA Panel on Biological Hazards
(BIOHAZ), 2010].
Despite extensive research, the natural routes of TSE transmission are
still rather poorly understood. Alimentary excretion and oral ingestion
contributes to horizontal transmission, but long incubation periods make it
difficult to link clinical cases to the original source of infection. Host
genetic factors, especially in sheep scrapie, make epidemiological
interpretations even more difficult, and the existence of different TSE strains
brings further complexity. Understanding the risks of transmission, especially
those posed by reproductive technologies, requires information on the presence
and levels of infectivity in various body tissues, which typically entails
detection of PrPSc by immunohistochemical or immunoblotting assays. However,
since the ability to detect PrPSc does not necessarily equate with the amount of
infectivity, bioassays in mice are also required.
The general conclusion from research to date is that the risk of TSE
infection by semen from sheep, goats and cattle is extremely low.
Cross-contamination of the semen samples used in this study is one explanation
for the infectivity detected in the semen samples from the infected animals.
However, TgSShpPrP mice injected with semen from the uninfected rams did not
show any sign of disease or the presence of PrPSc in their brains. The results
of the current study demonstrate detection of PASA in semen as well as the
presence of PrPSc in transgenic mice following IC injection. The detection of
PASA was performed by using normal brain homogenate as a PrPC source from sheep
with a genotype that, in some cases, did not match the genotype of the ram semen
samples. It is doubtful that this had any negative influence on PASA
amplification of the semen samples since previous studies using mismatched
genotypes similar to that for semen did not hinder amplification in blood and
urine (Rubenstein et al., 2011, 2011). The detection of the infectious agent in
semen by bioassays in ovinized transgenic mice contrasts with the previous
report by Sarradin et al. (2008). Sarradin et al. (2008) examined the role of
semen in sheep scrapie transmission by analysing whether semen (i.e. seminal
plasma and spermatozoa) could infect and cause disease in the tg338 transgenic
mouse line, which overexpresses the permissive V136R154Q171 allele of the ovine
prion protein. None of the tg338 mice injected with semen from the VRQ/VRQ
genotype or from the ARR/ARR control ram showed any clinical signs during the
period of the experiment. The semen samples differed between the two studies, as
did the transgenic mouse lines, and most likely the scrapie strain. While both
the VRQ and ARQ genotypes are more sensitive to scrapie infection than the ARR
genotype, it is conceivable that in the transgenic mouse bioassay the ARQ
protein confers greater susceptibility to semen-derived PrPSc. PrP in sperm is
known to undergo different processing than that in brain (Shaked et al., 1999;
Ecroyd et al., 2005). Further, the impact of alanine substitutions can be
substantial since the threonine to alanine mutation at position 183 of the human
prion protein is linked to a familial disease and results in retention of the
prion protein in the endoplasmic reticulum due to structural changes in the
protein (Capellari et al., 2000). Whether or not these factors were responsible
for the differences in the final outcomes of these studies remains to be
determined. However, it seems premature to rule out the potential of
semen-associated-seeding activity as a factor in the transmission of
scrapie.
Ram #8031 is of particular interest. He was euthanized due to an
overabundance of ARQ/ARQ and showed no clinical signs at the time of death at
age 16 months. All postmortem tissues were IHC negative (Table S2) and yet
sPMCA80 and sPMCA120 followed by SOFIA showed that PASA was present in the semen
of this ram. No doubt this ram was infected and subclinical. He had been born
and raised in a highly contaminated environment, but possessed a long incubation
time genotype. Typical of this strain, he would not have shown clinical signs or
positive testing utilizing IHC for 3–5 years. This supports and strengthens our
view that the assay used in this study is more sensitive than the presently
accepted IHC for early detection of scrapie and could be of value as a live
animal test.
We tend to believe that a positive ram was the likely source of the
scrapie-infected ewes, if not from semen, then by another type of contact. All
but one of the affected ewes were VRQ/VRQ; the incubation time of the ‘Caine’
strain of scrapie normally in these genotypes is 4–7 months (Bulgin et al.,
2006; Hamir et al., 2009), suggesting that the exposure occurred during the
period when the rams were placed with the ewes for breeding. However, the
presence of the infectious agent in the semen of the 136AV and VV rams infected
with the ‘Caine’ strain of scrapie, adds another intriguing possibility.
An interesting question is why it took so long for the disease to present
itself when the first exposure of the flock during breeding to positive rams
occurred 4 years previously. During the preceding 7 years, even though the flock
was genetically susceptible, there was no scrapie transmission. The disease
onslaught definitely appears to be promoted by the change in genetic composition
of the sentinel flock and their degree of susceptibility. When the 136VV and AV
rams were first introduced to the flock in 2004, the ewes were all 136AA. Thus,
the population slowly shifted to become more susceptible as the 136V genotype
was selected for. It is noteworthy that every one of the 136VV ewes in the flock
(26 %) was affected. Only one 136AV ewe was affected in spite of the fact that
AV ewes made up the majority of the flock (57.9 %). Interestingly, of the VV
ewes, two had been in the flock for 3 years, two for 2 years and three were
yearlings.
In summary, it seems premature to rule out the potential of
semen-associated-seeding activity as a factor in transmission of scrapie. In
that regard, experiments are currently under way to assess the transmissibility
of the PrPSc from the clinically normal mice that had been infected with sheep
semen as well as the material produced by PMCA using a semen seed. These results
will demonstrate whether the seeding activity found in semen is competent to
promote scrapie transmission. Also, demonstration of natural transmission of
scrapie in sheep by semen will have to be confirmed by inoculation of highly
susceptible oestrous ewes via artificial insemination using semen collected from
positive ‘Caine’ strain scrapie-infected rams since this may be a
strain-specific phenomenon.
PrPSc has been detected in sheep, deer, and hamster blood, both at
terminal stages of disease and in pre-symptomatic animals [21], [36], [45]–[48]
and in CSF and urine from naturally and experimentally infected animals [22],
[34], [48]–[50]. Furthermore, epidemiologic studies have demonstrated the
presence of prion infectivity in blood and plasma from vCJD cases [51]–[54]. It
is possible that the level of infectivity and/or PrPSc is too low to be detected
or that there may be various forms of sCJD with subtle differences, which
reflects varied amounts of infectivity in the blood. We have shown that PrPSc is
detectable by western blotting of samples from CNS and lymphoid tissues,
including tonsil, of both vCJD and sCJD patients. We also demonstrated that
PrPSc is detectable by SOFIA in unconcentrated, and even highly diluted, CNS and
non-CNS tissues from human prion disease samples, as well as in CSF from sCJD
patients. Thus, our assay could be used for antemortem diagnosis of both sCJD
and vCJD. In addition, our data suggests that PMCA of blood followed by SOFIA
should be pursued as a non-invasive assay for antemortem diagnosis of vCJD. http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0066352
Sunday, August 02, 2015
TEXAS CWD, Have you been ThunderStruck, deer semen, straw bred bucks, super
ovulation, and the potential TSE Prion connection, what if?
Saturday, August 01, 2015
Texas CWD Medina Captive Two more deer test positive for chronic wasting
disease CWD TSE Prion
Sunday, August 02, 2015
TEXAS CWD, Have you been ThunderStruck, deer semen, straw bred bucks, super
ovulation, and the potential TSE Prion connection, what if?
Sunday, July 26, 2015
TEXAS IN MELT DOWN MODE OVER CAPTIVE CWD AND THEY ARE PUTTING LIPSTICK ON
THAT PIG AND TAKING HER TO THE DANCE LIKE MAD COW DISEASE
Thursday, July 23, 2015
Chronic Wasting Disease (CWD) 101 Drs. Walter Cook & Donald S. Davis
Tuesday, July 28, 2015
TEXAS Kills 35 Deer at Medina County Ranch (Texas Captive CWD)
Tuesday, July 21, 2015
*** Texas CWD Medina County Herd Investigation Update July 16, 2015
***
Friday, July 17, 2015
TPW Commission Holds Special Meeting on Chronic Wasting Disease
Wednesday, July 01, 2015
TEXAS Chronic Wasting Disease Detected in Medina County Captive Deer
Thursday, July 09, 2015
TEXAS Chronic Wasting Disease (CWD) Herd Plan for Trace-Forward Exposed
Herd with Testing of Exposed Animals
Tuesday, July 14, 2015
Texas Parks and Wildlife Commission Special Meeting Thursday on Chronic
Wasting Disease CWD
Rare report of deer disease in Texas causes stir
Houston Chronicle
Rare report of deer disease in Texas causes stir, especially since it’s the
8 case of CWD documented in Texas, and the first case of CWD in Captive deer.
here is how I would have titled this article, and why.
Shannon Tompkins Finally Breaks Silence on Texas First Captive CWD Case and
Starts Off Spreading False Information About Risk Factors. ...
Thursday, July 16, 2015
Wednesday, March 18, 2015
Chronic Wasting Disease CWD Confirmed Texas Trans Pecos March 18,
2015
Wednesday, March 25, 2015
Chronic Wasting Disease CWD Cases Confirmed In New Mexico 2013 and 2014
UPDATE 2015
Thursday, May 02, 2013
*** Chronic Wasting Disease (CWD) Texas Important Update on OBEX ONLY
TEXTING
Monday, February 11, 2013
TEXAS CHRONIC WASTING DISEASE CWD Four New Positives Found in Trans Pecos
Tuesday, July 10, 2012
Chronic Wasting Disease Detected in Far West Texas
Monday, March 26, 2012
Texas Prepares for Chronic Wasting Disease CWD Possibility in Far West
Texas
***for anyone interested, here is some history of CWD along the Texas, New
Mexico border, and my attempt to keep up with it...terry
snip...
see history CWD Texas, New Mexico Border ;
Monday, March 26, 2012
3 CASES OF CWD FOUND NEW MEXICO MULE DEER SEVERAL MILES FROM TEXAS BORDER
Sunday, October 04, 2009
CWD NEW MEXICO SPREADING SOUTH TO TEXAS 2009 2009 Summary of Chronic
Wasting Disease in New Mexico New Mexico Department of Game and Fish
Wednesday, August 05, 2015
*** Ohio confirms to me Chronic Wasting Disease CWD Spreads 19 confirmed
cases to date ***
Just got off the phone with Christy Clevenger of Ohio
Ohio Department of Agriculture March 2012 – Present (3 years 6
months)Reynoldsburg, Ohio CWD program
Ms. Clevenger confirmed, to date, from the Yoder debacle, 1 confirmed case
of CWD from the Hunting Preserve, 2 confirmed cases from the Breeding Farm, and
16 confirmed cases of CWD from the Breeder Depopulation, with a total to date of
19 cases of CWD in Ohio...
Terry
Thursday, August 06, 2015
Michigan DNR confirms third deer positive for CWD; hunter participation is
critical this fall
Tuesday, August 4, 2015
FDA U.S. Measures to Protect Against BSE
Diagnosis and Reporting of Creutzfeldt-Jakob Disease
Singeltary, Sr et al. JAMA.2001; 285: 733-734. Vol. 285 No. 6, February 14,
2001 JAMA
Diagnosis and Reporting of Creutzfeldt-Jakob Disease
Singeltary, Sr et al. JAMA.2001; 285: 733-734. Vol. 285 No. 6, February 14,
2001 JAMA
Diagnosis and Reporting of Creutzfeldt-Jakob Disease
To the Editor: In their Research Letter, Dr Gibbons and colleagues1
reported that the annual US death rate due to Creutzfeldt-Jakob disease (CJD)
has been stable since 1985. These estimates, however, are based only on reported
cases, and do not include misdiagnosed or preclinical cases. It seems to me that
misdiagnosis alone would drastically change these figures. An unknown number of
persons with a diagnosis of Alzheimer disease in fact may have CJD, although
only a small number of these patients receive the postmortem examination
necessary to make this diagnosis. Furthermore, only a few states have made CJD
reportable. Human and animal transmissible spongiform encephalopathies should be
reportable nationwide and internationally.
Terry S. Singeltary, Sr Bacliff, Tex
1. Gibbons RV, Holman RC, Belay ED, Schonberger LB. Creutzfeldt-Jakob
disease in the United States: 1979-1998. JAMA. 2000;284:2322-2323.
TSS
posted by Terry S. Singeltary Sr. at
6:30 PM
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