Progress 10/01/12 to 09/30/15
Outputs
Target Audience:The target audience for this work includes the prion research community, wildlife managers, the U.S. Environmental Protection Agency, state environmental agencies, state departments of public health, U.S. Department of Agriculture, andcrop industries in areas impacted by cervid chronic wasting disease and potentially bovine spongiform encephalopathy. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Annabel Austin, a DVM student at the University of Pennsylvania, completed a Merial Veterinary Summer Scholarship project focused on evaluating the oral transmissibility prion-contaminated plants. Ms. Austin carried out the PMCA analyses on rodent tissues to detect clinically and subclinically infected hosts resulting from oral consumption of prion-contaminated plants. C. Carlson, and A. Austin presented research findings from this project at national and international conferences. How have the results been disseminated to communities of interest?1.Carlson, C.M. Environmental prion transmission: Identifying novel disease dissemination routes. Invited presentation at the University of Wisconsin Women in Science Seminar Series. University of Wisconsin - River Falls, River Falls, WI (Feb 2015) 2. Johnson, C.J., C.M. Carlson, M.W. Keating, N.M. Gibbs, H. Chang, J.K. Wiepz, and J.A. Pedersen. Uptake of prions into plants.PRION 2015 Conference, Ft. Collins, CO (May 2015). 3. Carlson, C.M. Identifying novel environmental transmission routes for chronic wasting disease.Association of Fish and Wildlife Agencies & USGS Ecosystems Fish and Wildlife Disease Seminar Series. Online Webinar (July 2015). 4. Austin, A.G., C.M. Carlson, and C.J. Johnson. Evaluation of the transmissibility of prions in plants.Merial-NIH National Veterinary Scholars Symposium. University of California - Davis, Davis, CA (August 2015). 5. Pedersen, J.A. Uptake of prions into plants and inactivation of prions. Huazhong Agricultural University, Wuhan, China, (December 2015). What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The amount of infectious prion protein taken up by the model plantArabidopsis thalianawas determined taking into account inhibitory effects of plant tissue on protein misfolding cyclic amplification (PMCA), the assay used to amplify, detect and quantify prion protein in plant samples. PMCA analyses of brain and peripheral tissues from rodents orally challenged with prion contaminated plants were used to confirm prion disease in clinical animals and to identify sub-clinically infected animals. Additional procedural controls were performed on our hydroponic, agar, and soil growth systems. We also initiated the study of the uptake of the fluorescent proteinmCherry viaArabidopsis thaliana roots to further examine mechanisms of intact protein uptake by plants. Lastly, one manuscript is being finalized further.
Publications
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Progress 10/01/13 to 09/30/14
Outputs
Target Audience: Prion research community, state government officials. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? The M.S. student involved in the project defended his thesis titled "Assessment of Prion Uptake by Plants using Protein Misfolding Cyclic Amplification" in partial fulfillment of the requirements for his degree in Civil and Environmental Engineering. Both the M.S. and Ph.D. students involved in the project trained laboratory staff to conduct the novel microplate-based PMCA technique. How have the results been disseminated to communities of interest? A collaborating scientist (C. Johnson) on the project presented results from this study as an invited speaker at an international prion conference. The project director communicated results to state governmental officials. What do you plan to do during the next reporting period to accomplish the goals? During the next reporting period, we intend to publish our findings on prion uptake and quantitation in various species of plants and from various types of media (hydroponic, agar, and soil) as well as those indicating oral transmissibility of prions taken up by plants to rodent hosts. We plan to conduct further experiments to elucidate the mechanisms of prion uptake by plants.
Impacts
What was accomplished under these goals?
The amount of infectious prion protein taken up by the model plantArabidopsis thalianaand several crop species (alfalfa, barley, maize) grown on hydroponic, agar, and soil media was quantified using protein misfolding cyclic amplification (PMCA) technology. This was accomplished by adaptation and optimization of a new, high throughput and more efficient PMCA method over previous methods used, termed microplate-based PMCA (Moudjou et al. 2013. MBio. 5(1):e00829-13), to the detection of prion protein taken up by plants in our studies. Knowledge of the quantity of infectious prion protein per weight of plant tissue allows us to make initial assessments of risk of infection to mammalian hosts posed by consumption of prion contaminated plants. Rodent bioassays in which animals were orally inoculated with prion contaminated plants and assessed for disease transmission were completed during this reporting period.
Publications
- Type:
Theses/Dissertations
Status:
Submitted
Year Published:
2014
Citation:
Keating, M.W. Assessment of Prion Uptake by Plants using Protein Misfolding Cyclic Amplification. M.S. dissertation, University of Wisconsin, Madison, USA, 2014
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? The Ph.D. student involved in the project participated in an international prion conference where she won an award for the best poster. The Ph.D. student involved in the project trained a M.S. student to conduct analyses for prions. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals? During the next reporting period, we intend to publish our initial findings, conduct further experiments to assess prion uptake from soil to plants, and begin examining mechanisms of uptake.
Impacts
What was accomplished under these goals?
To determine plant uptake and transmission of infectious prion protein, we exposed the model plant Arabidopsis thaliana and several crop species (alfalfa, barley, maize, tomato) to prion-contaminated hydroponic, agar, and soil media and tested for the presence of prion protein and infectivity by confocal laser scanning microscopy (CLSM), protein misfolding cyclic amplification with beads (PMCAb) and rodent bioassay. CLSM microscopy was used to assess uptake of fluoresecently labeled pathogenic prion protein into roots. PMCAb and rodent bioassays (intracerebral and oral routes of inoculation) were used to assess the translocation of prion infectivity to aerial portions of plants (i.e., leaves, stems).
Publications
- Type:
Theses/Dissertations
Status:
Submitted
Year Published:
2013
Citation:
Carlson, C.M. Environmental prion transmission: Identifying novel prion dissemination routes. Ph.D. dissertation, University of Wisconsin, Madison, USA, 2013
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: This project began in October. We have set up plant exposures to prions. We have adapted our detection method method for prion protein in plant extracts. PARTICIPANTS: Joel A. Pedersen (principal investigator) - overall supervision of project; Clarissa Booth (graduate research assistant) - trained prospective (rotating) graduate student on techniques relevant for project; Christina Carlson (graduate research assistant) - initiated plant exposures, optimized detection method; Christopher J. Johnson (collaborator, USGS National Wildlife Health Center) - supervision of activities at his facility related to project; Haeyoon Chang (biological science technician, USGS National Wildlife Health Center) - provided laboratory assistance to Carlson. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Conditions have been identified to allow successful detection of pathogenic prion protein in plant extracts.
Publications
- No publications reported this period
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