DETECTION OF PrPCWD IN FECES FROM NATURALLY EXPOSED ROCKY MOUNTAIN ELK (CERVUS ELAPHUS NELSONI) USING PROTEIN MISFOLDING CYCLIC AMPLIFICATION
DETECTION OF PrPCWD IN FECES FROM NATURALLY EXPOSED ROCKY MOUNTAIN ELK (CERVUS ELAPHUS NELSONI) USING PROTEIN MISFOLDING CYCLIC AMPLIFICATION
Bruce Pulford1, Terry R. Spraker1, A. Christy Wyckoff1, Crystal Meyerett1, Heather Bender1, Adam Ferguson1, Brittney Wyatt1, Krista Lockwood1, Jenny Powers2, Glenn C. Telling1, Margaret A. Wild2 and Mark D. Zabel1,3
+ Author Affiliations
1Department of Microbiology, Immunology and Pathology, Prion Research Program, Colorado State University, 1619 Campus Delivery, Fort Collins, Colorado 80523, USA
2National Park Service, 1201 Oakridge Drive, Fort Collins, Colorado 80525, USA
↵3 Corresponding author (email: mark.zabel@colostate.edu)
Abstract
Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy affecting captive and free-ranging cervids. Currently, tests for CWD in live animals involve relatively invasive procedures to collect lymphoid tissue biopsies and examine them for CWD-associated, protease-resistant cervid prion protein (PrPCWD) detected by immunohistochemistry (IHC). We adapted an ultrasensitive prion detection system, protein misfolding cyclic amplification (PMCA), to detect PrPCWD in Rocky Mountain elk (Cervus elaphus nelsoni) feces. Our PMCA reproducibly detected a 1.2×107 dilution of PrPCWD (a 10% infected brain homogenate diluted 1.2×106-fold into 10% fecal homogenates), equivalent to approximately 100 pg of PrPCWD/g of feces. We developed a semiquantitative scoring system based on the first PMCA round at which PrPCWD was detected and fit a nonlinear regression curve to our serial dilutions to correlate PMCA scores with known PrPCWD concentrations. We used this PMCA scoring system to detect PrPCWD and estimate its concentration in feces from free-ranging elk from Rocky Mountain National Park, Colorado. We compared our results to PrPCWD IHC of rectoanal mucosa-associated lymphoid tissue and obex from the same animals. The PMCA successfully detected PrPCWD in feces from elk that were positive by IHC, with estimated prion loads from 100 to 5,000 pg PrPCWD/g of feces. These data show for the first time PrPCWD in feces from naturally exposed free-ranging elk and demonstrate the potential of PMCA as a new, noninvasive CWD diagnostic tool to complement IHC.
http://www.jwildlifedis.org/content/48/2/425.abstract?etoc
Survival Patterns in White-tailed and Mule Deer after Oral Inoculation with a Standardized, Conspecific Prion Dose
Michael W. Miller1,4, Lisa L. Wolfe1, Tracey M. Sirochman1, Michael A. Sirochman1, Jean E. Jewell2 and Elizabeth S. Williams2,3
+ Author Affiliations
1Colorado Division of Wildlife, Wildlife Research Center, 317 West Prospect Road, Fort Collins, Colorado 80526-2097, USA
2Wyoming State Veterinary Laboratory, Department of Veterinary Sciences, University of Wyoming, 1174 Snowy Range Road, Laramie, Wyoming 82070, USA
↵4 Corresponding author (email: mike.miller@state.co.us)
Abstract
We orally inoculated white-tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus) with a standardized, conspecific prion dose and collected biologic samples throughout the disease course. Mule deer (PRNP genotype 225SS) and PRNP genotype 96GG white-tailed deer succumbed along similar trajectories, but 96GS- and 96SS-genotype individuals tended to survive longer. Received September 8, 2011. Accepted November 23, 2011. © Wildlife Disease Association 2012
http://www.jwildlifedis.org/content/48/2/526.abstract?etoc
Assessment of Prospective Preventive Therapies for Chronic Wasting Disease in Mule Deer
Lisa L. Wolfe1,4, David A. Kocisko2,3, Byron Caughey2 and Michael W. Miller1
+ Author Affiliations
1Colorado Division of Wildlife, Wildlife Research Center, 317 West Prospect Road, Fort Collins, Colorado 80526-2097, USA
2Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840, USA
↵4 Corresponding author (email: lisa.wolfe@state.co.us)
Abstract
We compared prion infection rates among mule deer (Odocoileus hemionus) receiving pentosan polysulfate, tannic acid, tetracycline HCl, or no treatment 14 days before to 14 days after (dpi) oral inoculation with tonsil tissue homogenate. All deer were infected, but the rapid disease course (230–603 dpi) suggested our challenge was overwhelming. Received September 8, 2011. Accepted November 23, 2011. © Wildlife Disease Association 2012
http://www.jwildlifedis.org/content/48/2/530.abstract?etoc
Thursday, April 05, 2012
Prevalence and Effects of Chronic Wasting Disease in Elk from Rocky Mountain National Park
http://chronic-wasting-disease.blogspot.com/2012/04/prevalence-and-effects-of-chronic.html
http://chronic-wasting-disease.blogspot.com/
TSS
Bruce Pulford1, Terry R. Spraker1, A. Christy Wyckoff1, Crystal Meyerett1, Heather Bender1, Adam Ferguson1, Brittney Wyatt1, Krista Lockwood1, Jenny Powers2, Glenn C. Telling1, Margaret A. Wild2 and Mark D. Zabel1,3
+ Author Affiliations
1Department of Microbiology, Immunology and Pathology, Prion Research Program, Colorado State University, 1619 Campus Delivery, Fort Collins, Colorado 80523, USA
2National Park Service, 1201 Oakridge Drive, Fort Collins, Colorado 80525, USA
↵3 Corresponding author (email: mark.zabel@colostate.edu)
Abstract
Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy affecting captive and free-ranging cervids. Currently, tests for CWD in live animals involve relatively invasive procedures to collect lymphoid tissue biopsies and examine them for CWD-associated, protease-resistant cervid prion protein (PrPCWD) detected by immunohistochemistry (IHC). We adapted an ultrasensitive prion detection system, protein misfolding cyclic amplification (PMCA), to detect PrPCWD in Rocky Mountain elk (Cervus elaphus nelsoni) feces. Our PMCA reproducibly detected a 1.2×107 dilution of PrPCWD (a 10% infected brain homogenate diluted 1.2×106-fold into 10% fecal homogenates), equivalent to approximately 100 pg of PrPCWD/g of feces. We developed a semiquantitative scoring system based on the first PMCA round at which PrPCWD was detected and fit a nonlinear regression curve to our serial dilutions to correlate PMCA scores with known PrPCWD concentrations. We used this PMCA scoring system to detect PrPCWD and estimate its concentration in feces from free-ranging elk from Rocky Mountain National Park, Colorado. We compared our results to PrPCWD IHC of rectoanal mucosa-associated lymphoid tissue and obex from the same animals. The PMCA successfully detected PrPCWD in feces from elk that were positive by IHC, with estimated prion loads from 100 to 5,000 pg PrPCWD/g of feces. These data show for the first time PrPCWD in feces from naturally exposed free-ranging elk and demonstrate the potential of PMCA as a new, noninvasive CWD diagnostic tool to complement IHC.
http://www.jwildlifedis.org/content/48/2/425.abstract?etoc
Survival Patterns in White-tailed and Mule Deer after Oral Inoculation with a Standardized, Conspecific Prion Dose
Michael W. Miller1,4, Lisa L. Wolfe1, Tracey M. Sirochman1, Michael A. Sirochman1, Jean E. Jewell2 and Elizabeth S. Williams2,3
+ Author Affiliations
1Colorado Division of Wildlife, Wildlife Research Center, 317 West Prospect Road, Fort Collins, Colorado 80526-2097, USA
2Wyoming State Veterinary Laboratory, Department of Veterinary Sciences, University of Wyoming, 1174 Snowy Range Road, Laramie, Wyoming 82070, USA
↵4 Corresponding author (email: mike.miller@state.co.us)
Abstract
We orally inoculated white-tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus) with a standardized, conspecific prion dose and collected biologic samples throughout the disease course. Mule deer (PRNP genotype 225SS) and PRNP genotype 96GG white-tailed deer succumbed along similar trajectories, but 96GS- and 96SS-genotype individuals tended to survive longer. Received September 8, 2011. Accepted November 23, 2011. © Wildlife Disease Association 2012
http://www.jwildlifedis.org/content/48/2/526.abstract?etoc
Assessment of Prospective Preventive Therapies for Chronic Wasting Disease in Mule Deer
Lisa L. Wolfe1,4, David A. Kocisko2,3, Byron Caughey2 and Michael W. Miller1
+ Author Affiliations
1Colorado Division of Wildlife, Wildlife Research Center, 317 West Prospect Road, Fort Collins, Colorado 80526-2097, USA
2Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840, USA
↵4 Corresponding author (email: lisa.wolfe@state.co.us)
Abstract
We compared prion infection rates among mule deer (Odocoileus hemionus) receiving pentosan polysulfate, tannic acid, tetracycline HCl, or no treatment 14 days before to 14 days after (dpi) oral inoculation with tonsil tissue homogenate. All deer were infected, but the rapid disease course (230–603 dpi) suggested our challenge was overwhelming. Received September 8, 2011. Accepted November 23, 2011. © Wildlife Disease Association 2012
http://www.jwildlifedis.org/content/48/2/530.abstract?etoc
Thursday, April 05, 2012
Prevalence and Effects of Chronic Wasting Disease in Elk from Rocky Mountain National Park
http://chronic-wasting-disease.blogspot.com/2012/04/prevalence-and-effects-of-chronic.html
Saturday, April 07, 2012
EFFECTS OF CHRONIC WASTING DISEASE ON REPRODUCTION AND FAWN HARVEST VULNERABILITY IN WISCONSIN WHITE-TAILED DEER
http://chronic-wasting-disease.blogspot.com/
TSS
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