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United States Department of Agriculture

Agricultural Research Service

2007 Annual Report

1a.Objectives (from AD-416)
1. We will determine enteric dose-response relationships for crude preparations of botulinum neurotoxin (BoNT)in food using rodent models. 2. We will optimize sample preparation and develop rapid immunological and biochemical tests for BoNT.

1b.Approach (from AD-416)
1. We will administer BoNT in buffer and BoNT spiked food to mice, testing various time and temperature storage conditions (including heating) for effects on toxicity. We will compare commercially available purified BoNT to crude toxin (sterile culture fractions). 2. We will evaluate BoNT assays currently available commercially or through collaborators. We will develop new monoclonal antibodies to BoNT toxin and toxoid. Initial evaluations will be made in ELISA and other standard formats, and selected assays will be ported onto new assay platforms. Examples of new platforms include microbead assays read via microfluorimetry, paramagnetic fluorescent nanosphere assays, microarray analyses employing glass or nitrocellulose surfaces, "dip-stick" format rapid ELISAs, and "black-box" turnkey assay systems. Biochemical assays will include measurements of protease activity using fluorescence polarization and/or fluorescence resonance energy transfer. Documents SCA with the University of Wisconsin-Madison.

3.Progress Report
This report documents research conducted under a Specific Cooperative Agreement between ARS and the University of Wisconsin. Additional details of research can be found in the report for the parent CRIS 5325-42000-043-00D, Development of Detection Technologies for Toxins and Their Validation in Food Matrices. In FY07 this project was extended into a 2d year, albeit with no additional funding.

The original goal of the research was collaborative development of sample preparation and assay technologies for detection of botulinum neurotoxin in foods. However, because of shifting priorities on the part of the Cooperator, the SCA has served mostly to provide advice on administration of Select Agent research and regular delivery of critical crude and purified botulinum neurotoxin reagents to the ADODR. These services are valuable and cost-effective.

The ADODR monitors this project through occasional telephone conversations and frequent email and fax exchanges with the Cooperator. The ADODR’s staff and the Cooperator and staff occasionally meet at national scientific meetings for updates and additional coordination. All material shipments were documented and contents confirmed upon arrival.

Because of administrative delays work under the SCA achieved a late start. However, it is now proceeding well. We presented several collaborative abstracts in FY07.

Last Modified: 8/4/2015
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