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
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
This report documents research conducted under a Reimbursable Agreement between ARS and the UNIVERSITY OF WISCONSIN. The research has included collaborative development of assay technologies for detection of botulinum neurotoxins in foods. The Cooperator also provides advice on administration of Select Agent research and provides delivery of critical crude and purified botulinum neurotoxin reagents to the ADODR. These services have been valuable and cost-effective. All material shipments were documented and contents confirmed upon arrival.
The ADODR monitors this project through telephone conversations, email, and fax exchanges with the Cooperator. The ADODR’s staff and the Cooperator meet at technical meetings and have jointly prepared publications.