Monday, January 26, 2009

Chronic Wasting Disease cases up for second year South Dakota Game, Fish & Parks Department As of 12-31-2008

Chronic wasting cases up for second year By Journal staff Sunday, January 25, 2009

State wildlife biologists say tests for the second year found more cases of a fatal brain disease in deer and elk shot in the state. Big-game seasons ended Jan. 20.

Chronic wasting disease is caused by an abnormal protein, causing the animals to increase water consumption but lose weight and muscle control. The disease is not contagious to humans, but as a precaution, health officials advise to eat only muscle meat from healthy elk and deer, and not to eat any meat from animals that showed signs of the disease or test positive for CWD. Hunters also are urged to take precautions in field dressing the carcasses.

The South Dakota Game, Fish & Parks Department says hunters provided 2,052 elk, white-tailed deer and mule deer carcasses for testing, with 27 specimens showing CWD. Last year, 19 cases were found among 2,558 tests.

Big game biologist Steve Griffin in a news release said most of the cases came from areas with previous CWD cases, although “a couple of new areas” now apparently are contaminated. The release did not disclose the new locations.

Almost every state wildlife agency now conducts CWD testing. The disease is known to infect free-ranging and ranch-raised deer and elk in areas of 13 states and two Canadian provinces.



As of 12-31-2008

1,895 total samples collected since July 1, 2008:

380 elk 419 mule deer 1,096 white-tailed deer Most samples are from the Black Hills and from prairie hunting units in Fall River, Custer, and eastern Pennington counties of western South Dakota. Most samples were taken from hunter-harvested animals. RESULTS As of 12-31-2008 we have received results from the SDSU Diagnostic Lab or Wind Cave National Park on 1,800 samples listed below.

380 elk 412 mule deer 1,008 white-tailed deer

Of the 1,800 samples tested to date, we have found 9 CWD positive Elk and 17 CWD positive deer. Below is a summary of these animals:

Elk female from Wind Cave National Park in Custer County. (Sick/Surveillance) Elk female from Unit H3B in Custer County. (Hunter Harvest) Elk male from Unit H3A in Custer County. (Hunter Harvest) Elk female from Wind Cave National Park in Custer County. (Sick/Surveillance) Elk male from Unit H3A in Custer County. (Hunter Harvest) White-tailed female from Unit BD4 in Custer County. (Sick/Surveillance) Elk female from Unit H3C in Fall River County. (Hunter Harvest) Elk female from Wind Cave National Park in Custer County. (Sick Surveillance) White-tailed female from Unit 27A in Custer County. (Hunter Harvest Mule deer male from Unit 27B in Fall River County. (Hunter Harvest)

Mule deer female from Unit 21B in Custer County. (Hunter Harvest)

White-tailed female from Unit 27B in Fall River County. (Hunter Harvest)

White-tailed male from Unit BD3 in Pennington County. (Sick/Surveillance)

Mule deer male from Unit 27B in Fall River County. (Hunter Harvest)

Mule deer female from Unit 27B in Fall River County. (Hunter Harvest)

Mule deer female from Unit 21B in Custer County. (Hunter Harvest)

Elk male from Wind Cave National Park in Custer County. (Sick Surveillance)

Mule deer male from Unit 27B in Fall River County. (Hunter Harvest)

White-tailed male from Unit 21A in Custer County. (Hunter Harvest)

Mule deer female from Unit 27A in Fall River County. (Hunter Harvest) Mule deer female from Unit 21A in Custer County. (Hunter Harvest)

Mule deer female from Unit 27B in Fall River County. (Hunter Harvest)

White-tailed male from Unit BH1 in Custer County. (Hunter Harvest)

Elk female from Unit H3C in Custer County. (Hunter Harvest)

Mule deer female from Hot Springs in Fall River County. (City Deer Removal)

Mule deer female from Hot Springs in Fall River County. (City Deer Removal)

In Summary: South Dakota Department of Game Fish and Parks and Wind Cave National Park has found 26 case of CWD (9 elk, 17 deer) in free ranging cervids in the testing period -July 1, 2008 to present.

To date, South Dakota has found 103 cases of CWD (71 deer and 32 elk) in free ranging deer and elk since testing began in 1997. Wind Cave National Park accounts for 24of these animals (16 elk, 8 deer). A total of 19,299 wild deer and elk have been tested for CWD since 1997.

Hunters may get their animal tested for chronic wasting disease by making their own arrangements directly through the SDSU Diagnostic Lab at (605) 688-5171.


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Research Project: Detection of Transmissible Spongiform Encephalopathy Agents in Livestock, Wildlife, Agricultural Products, and the Environment Location: Foodborne Contaminants Research

2008 Annual Report

1a.Objectives (from AD-416) We will develop highly sensitive diagnostic tests to detect transmissible spongiform encephalopathy (TSE) in livestock and wildlife animal species prior to the onset of clinical disease. We will also develop tests to confirm the presence or absence of TSE disease agents in ingredients of animal origin and decontaminated environments.

1b.Approach (from AD-416) The threat of BSE continues to affect export economics for US meat. Meanwhile scrapie continues to influence sheep profits and herd biosecurity, and CWD is spreading throughout North America. Thus U.S. animal industry stakeholders have identified detection of the TSE infectious agent (prions) as a priority biosecurity research issue essential for prevention of TSE diseases. We will build on our previous successes using mass spectrometry (MS) for high-sensitivity and specificity in detection of PrPsc as a marker for TSE infectivity in blood using a hamster scrapie model. We will also develop a novel PrP-null mouse strain and related myeloma cell culture system for production of monoclonal antibodies (MAb), which may be specific for PrPsc. We will then choose MS or MAb and validate our novel diagnostic for preclinical diagnosis of scrapie in sheep blood. Whereas MS and MAb methods rely on dissolved samples, contamination of agricultural products and environmental surfaces is associated with solid samples. So we will produce a cell culture based assay for TSE infectivity that is surface-adsorbed. After using the relatively convenient hamster model for early development, we will validate this technology for detection of scrapie in sheep brain on meat-and-bone meal and stainless steel. Replacing 5325-32000-007-00D (3/19/2008).

3.Progress Report At this point in the Project, in general, we are completing preliminary studies using our relatively convenient hamster and mouse models, and are starting to work with more agriculturally relevant sheep and deer tissues. We are finding the cervid tissues quite different from rodent tissues, in their requirements for sample workup (e.g., amount and quality of lipid and fiber) and in their expression of TSE infectivity and presence of markers. OSQR required us to establish a new collaboration with a reputable cell biologist, to assist with our cell-based scrapie assay. We now have a new MTA with Dr. Charles Weissmann (Scripps), under which we are sharing cell lines and laboratory protocols. We have completed one part of our speed congenics project to develop PrP-null (disease-resistant) mice for use in antibody generation. After conceiving a new procedure for immunogen enrichment, we performed experimental vaccination of these animals in our facilities. This project relates to NP103 Component 8: Prevention and control of transmissible spongiform encephalopathies. Problem statement 9A: Scrapie; 9B Chronic Wasting Disease (CWD); and 9C: Bovine Spongiform Encephalopathy (BSE).

4.Accomplishments 1. Proteinase K-free method for preparation of samples facilitates TSE blood assay.

The most widely used and regulatory approved methods for detection of Transmissible Spongiform Encephalopathy (TSE) contain a step in which the sample is subjected to digestion by a very strong enzyme, proteinase K, which degrades almost all proteins in the sample except for an Infectious isoform of the normal cellular prion protein, a prion (PrPsc). Although PrPsc has served well as a marker for brain disease, infectivity in the blood is mostly not proteinase K resistant. The proteinase K-free technique developed by ARS scientists in the Foodborne Contaminants Research Unit in Albany, CA will allow scientists to detect infectivity in blood. These efforts will lead to diagnostic tests that will save farmers and ranchers money and resources by allowing them to identify infected animals prior to purchase, sale or slaughter, and keep TSE-infected animals out of the US food supply. This accomplishment addresses NP103 Component 8: Prevention and Control of Transmissible Spongiform Encephalopathies; Problem Statement 9A: Scrapie; 9B: Chronic Wasting Disease (CWD); and 9C: Bovine Spongiform Encephalopathy (BSE).

2. Demonstrated conversion of a non-infectious normal cellular prion protein (PrP) into disease isoform in cell culture.

Although Transmissible Spongiform Encephalopathy (TSE) infectivity can be detected using animal models and mass spectroscopy, a cell culture system offers increased speed and throughput. ARS scientists in the Foodborne Contaminants Research Unit in Albany, CA developed conditions for growth and infection of existing cell cultures and cultures expressing transgenic PrP genes, observing conversion to the disease-associated PrPsc isoform. This method will be further developed to detect infectivity that is adsorbed onto surfaces, such as stainless steel and soil. These efforts will lead to diagnostic tests that will save farmers and ranchers money and resources by allowing them to identify infected areas and equipment before these areas or items can infect their animals. This accomplishment addresses NP103 Component 8: Prevention and Control of Transmissible Spongiform Encephalopathies; Problem Statement 9A: Scrapie; 9B: Chronic Wasting Disease (CWD); and 9C: Bovine Spongiform Encephalopathy (BSE).

5.Significant Activities that Support Special Target Populations None.

6.Technology Transfer Number of New Commercial Licenses Executed 1

Review Publications Bruederle, C.E., Hnasko, R.M., Kraemer, T., Garcia, R.A., Haas, M.J., Marmer, W.N., Carter, J.M. 2008. Prion infected Meat-and-Bone Meal is still infectious after biodiesel production. PLoS Pathogens. Available:

Onisko, B.C., Chen, N., Napoli, J. 2008. The Nuclear Transcription Factor RAR Associates with Neuronal RNA Granules and Suppresses Translation. Journal of Biological Chemistry. 283(30):20841-20847.

Sajnani, G., Pastrana, M.A., Dynin, I.A., Onisko, B.C., Requena, J.R. 2008. Insights on scrapie prion protein (prpsc) structure obtained by limited proteolysis and mass spectrometry. Journal of Molecular Biology. 382(2008):88-98.

FY2006: Tests for prion contamination in soil and water will be developed.


Scrapie Agent (Strain 263K) Can Transmit Disease via the Oral Route after Persistence in Soil over Years Bjoern Seidel1#*, Achim Thomzig2#, Anne Buschmann3#, Martin H. Groschup3, Rainer Peters1, Michael Beekes2, Konstantin Terytze4

1 Fraunhofer Institute for Molecular Biology und Applied Ecology (IME), Schmallenberg, Germany, 2 P24 -Transmissible Spongiform Encephalopathies, Robert Koch-Institut, Berlin, Germany, 3 Institute for Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Insel Riems, Germany, 4 German Federal Environmental Agency (Umweltbundesamt, UBA), Dessau, Germany

Abstract The persistence of infectious biomolecules in soil constitutes a substantial challenge. This holds particularly true with respect to prions, the causative agents of transmissible spongiform encephalopathies (TSEs) such as scrapie, bovine spongiform encephalopathy (BSE), or chronic wasting disease (CWD). Various studies have indicated that prions are able to persist in soil for years without losing their pathogenic activity. Dissemination of prions into the environment can occur from several sources, e.g., infectious placenta or amniotic fluid of sheep. Furthermore, environmental contamination by saliva, excrements or non-sterilized agricultural organic fertilizer is conceivable. Natural transmission of scrapie in the field seems to occur via the alimentary tract in the majority of cases, and scrapie-free sheep flocks can become infected on pastures where outbreaks of scrapie had been observed before. These findings point to a sustained contagion in the environment, and notably the soil. By using outdoor lysimeters, we simulated a contamination of standard soil with hamster-adapted 263K scrapie prions, and analyzed the presence and biological activity of the soil-associated PrPSc and infectivity by Western blotting and hamster bioassay, respectively. Our results showed that 263K scrapie agent can persist in soil at least over 29 months. Strikingly, not only the contaminated soil itself retained high levels of infectivity, as evidenced by oral administration to Syrian hamsters, but also feeding of aqueous soil extracts was able to induce disease in the reporter animals. We could also demonstrate that PrPSc in soil, extracted after 21 months, provides a catalytically active seed in the protein misfolding cyclic amplification (PMCA) reaction. PMCA opens therefore a perspective for considerably improving the detectability of prions in soil samples from the field.

Prions Adhere to Soil Minerals and Remain Infectious Christopher J. Johnson1,2, Kristen E. Phillips3, Peter T. Schramm3, Debbie McKenzie2, Judd M. Aiken1,2, Joel A. Pedersen3,4*

1 Program in Cellular and Molecular Biology, University of Wisconsin Madison, Madison, Wisconsin, United States of America, 2 Department of Animal Health and Biomedical Sciences, School of Veterinary Medicine, University of Wisconsin Madison, Madison, Wisconsin, United States of America, 3 Molecular and Environmental Toxicology Center, University of Wisconsin Madison, Madison, Wisconsin, United States of America, 4 Department of Soil Science, University of Wisconsin Madison, Madison, Wisconsin, United States of America

Abstract An unidentified environmental reservoir of infectivity contributes to the natural transmission of prion diseases (transmissible spongiform encephalopathies [TSEs]) in sheep, deer, and elk. Prion infectivity may enter soil environments via shedding from diseased animals and decomposition of infected carcasses. Burial of TSE-infected cattle, sheep, and deer as a means of disposal has resulted in unintentional introduction of prions into subsurface environments. We examined the potential for soil to serve as a TSE reservoir by studying the interaction of the disease-associated prion protein (PrPSc) with common soil minerals. In this study, we demonstrated substantial PrPSc adsorption to two clay minerals, quartz, and four whole soil samples. We quantified the PrPSc-binding capacities of each mineral. Furthermore, we observed that PrPSc desorbed from montmorillonite clay was cleaved at an N-terminal site and the interaction between PrPSc and Mte was strong, making desorption of the protein difficult. Despite cleavage and avid binding, PrPSc bound to Mte remained infectious. Results from our study suggest that PrPSc released into soil environments may be preserved in a bioavailable form, perpetuating prion disease epizootics and exposing other species to the infectious agent.

Synopsis Transmissible spongiform encephalopathies (TSEs) are a group of incurable diseases likely caused by a misfolded form of the prion protein (PrPSc). TSEs include scrapie in sheep, bovine spongiform encephalopathy ("mad cow" disease) in cattle, chronic wasting disease (CWD) in deer and elk, and Creutzfeldt-Jakob disease in humans. Scrapie and CWD are unique among TSEs because they can be transmitted between animals, and the disease agents appear to persist in environments previously inhabited by infected animals. Soil has been hypothesized to act as a reservoir of infectivity, because PrPSc likely enters soil environments through urinary or alimentary shedding and decomposition of infected animals. In this manuscript, the authors test the potential for soil to serve as a reservoir for PrPSc and TSE infectivity. They demonstrate that PrPSc binds to a variety of soil minerals and to whole soils. They also quantitate the levels of protein binding to three common soil minerals and show that the interaction of PrPSc with montmorillonite, a common clay mineral, is remarkably strong. PrPSc bound to Mte remained infectious to laboratory animals, suggesting that soil can serve as a reservoir of TSE infectivity.

Direct Detection of Soil-Bound Prions Sacha Genovesi1, Liviana Leita2, Paolo Sequi3, Igino Andrighetto4, M. Catia Sorgato1,5, Alessandro Bertoli1*

1 Dipartimento di Chimica Biologica, Università di Padova, Padova, Italy, 2 Istituto Sperimentale per la Nutrizione delle Piante, Gorizia, Italy, 3 Istituto Sperimentale per la Nutrizione delle Piante, Roma, Italy, 4 Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Italy, 5 CNR Istituto di Neuroscienze, Padova, Italy

Abstract Scrapie and chronic wasting disease are contagious prion diseases affecting sheep and cervids, respectively. Studies have indicated that horizontal transmission is important in sustaining these epidemics, and that environmental contamination plays an important role in this. In the perspective of detecting prions in soil samples from the field by more direct methods than animal-based bioassays, we have developed a novel immuno-based approach that visualises in situ the major component (PrPSc) of prions sorbed onto agricultural soil particles. Importantly, the protocol needs no extraction of the protein from soil. Using a cell-based assay of infectivity, we also report that samples of agricultural soil, or quartz sand, acquire prion infectivity after exposure to whole brain homogenates from prion-infected mice. Our data provide further support to the notion that prion-exposed soils retain infectivity, as recently determined in Syrian hamsters intracerebrally or orally challanged with contaminated soils. The cell approach of the potential infectivity of contaminated soil is faster and cheaper than classical animal-based bioassays. Although it suffers from limitations, e.g. it can currently test only a few mouse prion strains, the cell model can nevertheless be applied in its present form to understand how soil composition influences infectivity, and to test prion-inactivating procedures.

now, something i have pondered long about, with the atypical BSE in Texas and Alabama, where, as far as i know, those farms WERE NOT quarantined for 5 years due to an atypical TSE. HOWEVER, the farms of the atypical scrapie from where the mad sheep of mad river valley occurred, these farms were quarantined. ...

----- Original Message -----

From: Terry S. Singeltary Sr.

To: mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000214/!

Sent: Sunday, February 25, 2007 12:35 PM


Greetings USDA,

I respectfully request the final results of the mouse bio-assays test that were to have supposedly began 2+ years late, 5 years ago, on the imported sheep from Belgium ?

WHAT happened to the test results and MOUSE BIO-ASSAYS of those imported sheep from Belgium that were confiscated and slaughtered from the Faillace's, what sort of TSE did these animals have ?

WERE they atypical scrapie, BSE, and or typical scrapie ?

HOW much longer will you refuse to give us this information ? and for what reason ?

WHY is it that the Farm of the Mad Sheep of Mad River Valley were quarantined for 5 years, but none of these farms from Texas and Alabama with Atypical TSE in the Bovine, they have not been quarantined for 5 years,why not, with the real risk of BSE to sheep, whom is to say this was not BSE ?


full text ;

Monday, September 1, 2008


Greetings again BSE-L members,

I had a pleasant surprise this past Saturday. I got an unexpected package from O.I.G. on my old F.O.I.A. request, of the final test results of the infamous mad sheep of mad river valley. IF you all remember, back on Thu, 24 Apr 2008 15:00:20 -0500 I wrote ;

snip...full text ;;f=12;t=000469;f=12;t=000469

i remember a few years back ???

that a study showed the prion uptake in a tomato plant, not that this would surprise me ;

56. Members considered that there is no evidence that crops grown on the land which received composted excreta from BSE-challenged animals pose a TSE risk to humans or animals. One member suggested that, as some of these animals are orally challenged with high doses of BSE-infected materials, and the distribution of infectivity in the digestive system is not completely understood, it might be premature to conclude that there is no infective agent in the manure. Furthermore, an unpublished study had indicated low level absorption of PrP from soil by tomato plants although it should be noted that this study had not been repeated. Details of this work would be sent to the SEAC Secretary. Dr Matthews explained that most of the manure from animals challenged with high doses of BSE had already been composted and used for coppicing. Members agreed that the risks from disposal of residual manure from experimental animals would be much less than historic risks of on farm contamination from naturally infected animals at the height of the BSE epidemic.

disturbing to say the least. ...TSS

Tuesday, January 13, 2009

Antemortem detection of PrPCWD in preclinical, ranch-raised Rocky Mountain elk (Cervus elaphus nelsoni) by biopsy of the rectal mucosa Full Scientific Reports

Saturday, January 10, 2009

Chronic Wasting Disease Investigation Update Michigan December 18, 2008

Sunday, September 07, 2008

CWD LIVE TEST, and the political aspects or fallout of live testing for BSE in cattle in the USA

2008 CWD Laboratory Testing for Wild White-tailed Deer,1607,7-186-25806-202922--,00.html

Wednesday, January 07, 2009

CWD to tighten taxidermy rules Hunters need to understand regulations

Monday, January 05, 2009


Thursday, December 25, 2008

Lions and Prions and Deer Demise

Tuesday, January 06, 2009

CWD Update 93 December 29, 2008

Tuesday, September 09, 2008

CWD MICHIGAN UPDATE September 5, 2008

Monday, August 25, 2008


Saturday, January 24, 2009

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

8. A transgenic mouse for the study of chronic wasting disease (CWD). Environmental persistence of prions may be involved in transmission of CWD however there are few reliable biologic tools to assess whether water or soil samples contain CWD prions and to assess effectiveness of decontamination procedures. ARS scientists in the Animal Disease Research Unit in Pullman, WA, created a transgenic mouse in order to assess CWD contamination in a sample and determine by which routes in the body transmission may occur. The transgenic mouse is susceptible to CWD as indicated by neurologic symptoms and accumulation of abnormal prion protein, and testing is continuing to determine which routes of administration and how much sample can be used for detection of CWD prions. The mouse model has potential for use in detecting CWD infectivity in tissue and body fluids, in soil and water samples, and for assessing effectiveness of decontamination procedures. This work was performed in accordance with the ARS Animal Health National Program Component 8: Countermeasures to prevent and control transmissible spongiform encephalopathies (TSEs), addressing the problem of understanding transmission, infectivity, and pathogenesis.

5.Significant Activities that Support Special Target Populations None

6.Technology Transfer

Review Publications Schneider, D.A., Yan, H., Fry, L.M., Alverson, J., White, S.N., Orourke, K.I. 2008. Myenteric neurons of the ileum that express somatostatin are a target of prion neuroinvasion in an alimentary model of sheep scrapie. Acta Neuropathologica. 115(6):651-661.

Manning, L., Orourke, K.I., Knowles Jr, D.P., Marsh, S., Spencer, Y.I., Moffat, E., Wells, G.A., Czub, S. 2008. A collaborative Canadian-United Kingdom evaluation of an immunohistochemistry protocol to diagnose bovine spongiform encephalopathy. Journal of Veterinary Diagnostic Investigation. 20(4):113-117.

Hamir, A.N., Richt, J., Miller, J.M., Kunkle, R.A., Hall, S., Nicholson, E.M., Orourke, K.I., Greenlee, J.J., Williams, E.S. 2008. Experimental transmission of chronic wasting disease (CWD) of elk (Cervus elaphus nelsoni), white-tailed deer (Odocoileus virginianus), and mule deer (Odocoileus hemionus hemionus) to white-tailed deer by intracerebral route. Veterinary Pathology. 45(3):297-306.

O'Rourke, K.I., Spraker, T.R., Zhuang, D., Greenlee, J.J., Gidlewski, T.E., Hamir, A.N. 2007. Elk with a long incubation prion disease phenotype have a unique PrP-d profile. NeuroReport. 18(18):1935-1938.

Harrington, R.D., Baszler, T.V., Orourke, K.I., Schneider, D.A., Spraker, T.R., Liggitt, H.D., Knowles Jr, D.P. 2008. A species barrier limits transmission of chronic wasting disease to mink (Mustela vison). Journal of General Virology. 89(4):1086-1096.

Evoniuk, J., Johnson, M., Borowicz, P., Caton, J.S., Vonnahme, K., Reynolds, J., Taylor, J.B., Stoltenow, C., Orourke, K.I., Redmer, D. 2008. Effects of Nutrition and Genotype on Prion Protein (PrPC) Gene Expression in the Fetal and Maternal Sheep Placenta. Placenta. 29(5):422-428.

Evoniuk, J.M., Berg, P., Johnson, M.L., Larson, D.M., Maddock, T., Stoltenow, C.L., Schauer, C.S., Orourke, K.I., Redmer, D.A. 2007. Association between genotypes at codon 171 and 136 of the prion protein gene and production traits in market lambs. American Journal of Veterinary Research. 68(10):1073-1078.

Casas, E., White, S.N., Shackelford, S.D., Wheeler, T.L., Koohmaraie, M., Bennett, G.L., Smith, T.P.L. 2007. Assessing the association of single nucleotide polymorphisms at the thyroglobulin gene with carcass traits in beef cattle. Journal of Animal Science. 85:2807-2814.

Knowles Jr, D.P. 2008. Bovine Spongiform Encephalopathy. In: Brown, C., Torres, A., editors. The United States Animal Health Association - Foreign Animal Diseases. Seventh Edition. Boca Raton, FL: Boca Publications Group, Inc. p. 185-188.

Saturday, January 24, 2009

Bovine Spongiform Encephalopathy h-BSE ATYPICAL USA 2008 Annual Report Research Project: Study of Atypical Bse

Location: Virus and Prion Diseases of Livestock

2008 Annual Report

Terry S. Singeltary Sr. P.O. Box 42 Bacliff, Texas USA 77518



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