Graduate Thesis Or Dissertation

 

Improved detection methods and physiological studies of staphylococcal enterotoxin Public Deposited

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/5h73pz94v

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  • The purpose of this study was to develop means to facilitate the detection of staphylococcal enterotoxin and to elucidate factors that influence enterotoxin formation. It was found that semipurification flat-gel electrophoresis in a water-cooled apparatus was more efficient than CM-cellulose and Sephadex column chromatography currently used to separate toxin from soluble food constituents. The electrophoretic process was less time consuming, less expensive and adaptable to mulitple sample analyses. A technique combining isoelectric focusing in a polyacrylamidestabilized ampholyte solution followed by electro-osmophoresis was 5ucce ssful in reducing the time required for the separation of enteroioxii. l. A and the formation of a specific immuno-precipitate. It was possible by this method to assay enterotoxin A at a concentration of 0.5 μg /ml in five hours. Inhibitor studies were conducted on enterotoxin A and B production by Staphylococcus aureus 265-1 and 14458, respectively. The elaboration of enterotoxin A and B was inhibited by streptomycin sulfate, NaF, chioraniphenicol, KC1 and bacitracin. Inhibition by KC1 and NaF was reversed by Mg⁺⁺ . Penicillin G inhibited toxin production by S. aureus 14458 but not cell growth, whereas both toxin elaboration and growth were inhibited in studies with S. aureus 265-1, Enterotoxin B produced by S. aureus 1445 appeared during the latter part of the exponential growth phase. Nonreplicating cells at a concentration of 8 x 10¹⁰/ml were found capable of producing toxin in the presence of glucose and water alone. Chloramphenicol did not inhibit toxin formation in nonreplicating cells. Data suggested the existence of a toxin precursor pool and that toxin excretion may be an adjustment to static growth conditions.
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