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Title: IMMUNO-BASED METHODOLOGY FOR DETECTION OF MICROORGANISMS IN COMPETITIVE EXCLUSION CULTURES

Author
item Stanker, Larry
item Young, Colin
item Nisbet, David
item Deloach, John

Submitted to: Milk Food and Environmental Sanitarians International Association
Publication Type: Abstract Only
Publication Acceptance Date: 7/3/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: We previously described an anaerobic culture system (CF3) containing 29 bacterial species that were obtained from adult chickens. The administration of CF3 cultures to newly hatched chicks results in exclusion of Salmonella and is known as competitive exclusion. In order to evaluate the fate of these bacteria in the bird we have generated a panel of monoclonal antibodies against the constituent microorganisms of the mixed microflora CF3 culture. Presently, we have developed monoclonal antibodies to the following bacteria: Fusobacterium spp.; Bifidobacterium spp.; Enterococcus avium; Proprionobacteria granulosum; Eubacteria spp.; and Veillonella spp. from the CF3 culture. Data on the preparation and characterization of these monoclonal antibodies will be presented in this study. Five to ten individual monoclonal antibodies to each bacterial species listed above have been isolated. Isotype determination revealed antibodies of the following classes IgA, IgG1, IgG2a, IgG2b, IgG3, and IgM, and both kappa and lambda light chains. All of the antibodies were able to detect between 10**2 and 10**3 cfu. All displayed a high degree of specificity as determined by cross-reactivity to a panel of heterologous bacteria. Using these monoclonal antibodies we have developed a rapid method for detecting and quantifying the number of specific bacteria in the ceca of chickens and in the chemostats used to produce the CF3 material. The assay is formatted as a competition enzyme- linked immunosorbent assays (cELISA). Measurements of bacterial population in both the chemostat and in the ceca from chickens will be discussed.