In 1995, when we first tested the Clostridium chauvoei antiserum that serves as the raw material for our C. chauvoei conjugate, vehement expostulations proceeded from our darkened microscopy room that are probably not printable in this newsletter. The source of this angst was the fluorescence of not only rod-shaped C. chauvoeibacteria but also what appeared to be spirochetes. As you can see from Figures 1 & 2, there is a certain resemblance between the spiral structure that our conjugate labeled and a typical spirochete. We theorized that the C. chauvoei culture against which we had raised our antiserum (and from which we had made our slides) had been contaminated with a spirochete, resulting in the development of antibody not only to C. chauvoei but also to the mystery spirochete. Though we were somewhat chagrined by this development we decided that, since spirochetes are easy to differentiate from rods, the antibody would still be useful for detectingC. chauvoei. We conjugated the antibody, noted the apparent snafu on our Certificate of Analysis, and sold the conjugate.
Fast-forward a decade to 2005 when a gentleman named Peter Wragg at Veterinary Laboratory Agencies in the UK observed what we had thought was cross-reactivity. Being better informed than we, Peter knew that Clostridium chauvoei is peritrichous (has flagella uniformly distributed over its cell wall). Peter realized that what we had thought to be spirochetes were actually quite probably detached flagella. He was kind enough to share some photomicrographs of our conjugate staining a fresh culture of C. chauvoei that clearly show the flagella attached (Figure 3). Following Peter’s revelation, we applied our conjugate to quite a few wells of our C. chauvoei slides, which are made with our killed C. chauvoei culture, and photographed a number of spiral structures (Fig.4). We agree with Peter’s opinion of these structures and have modified our Certificate of Analysis accordingly.
We thank Peter Wragg for pointing out our error and kindly sharing his photomicrographs. We also thank Veterinary Laboratory Agencies for permitting the use of Peter’s findings and photos in this newsletter.
