2011 Annual Report 1a.Objectives (from AD-416) The objective of this cooperative research project is to develop an electrochemiluminescence assay for BoNT in foods, using novel monoclonal antibodies developed by our ARS laboratory. Specifically, BoNT occurs in many serotype variations. This Agreement addresses BoNT serotypes A, B, and E. The foods covered under this Agreement include ground beef, liquid eggs, milk, and leafy greens, such as lettuce and spinach. 1b.Approach (from AD-416) We will test various monoclonal antibodies developed by ARS, as well as additional monoclonal and polyclonal antibodies available through commercial license. Antibodies will be employed in pairs, in sandwich format, using the Meso Scale platform for electrochemiluminescence. For validation, crude supernatant from cultures of each serotype will be spiked into foods, and the Lower Limit Of Detection (LLOD) determined. 3.Progress Report We have successfully developed methods for detection of serotypes A and B of botulinum neurotoxin (BoNT) responsible for foodborne botulism, and we have moved on to look at serotype E. We also ported an enyzme-linked immunosorbent assay (ELISA) method for detection of ricin (the toxin from castor bean)to the electrochemiluminescence platform supported by this instrument. While the Cooperator has informally loaned us their instrument for over one year, we are negotiating terms for possible extension of the Agreement and the instrument loan, in order to participate with the Cooperator in an extension of this Agreement that would involve grant funding provided by a third party. The ADODR and staff consulted with the Cooperator via email, site visits by the Cooperator, and conference calls.