Graduate Thesis Or Dissertation

The lipids of spore types B, E, and F and vegetative cell types E and F of Clostridium botulinum

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  • The primary purpose of this investigation was to characterize the lipids of the spores and vegetative cells of Cl. botulinum. A second purpose was to explore the possibility that lipids might serve as a means of differentiating the chiefly proteolytic Cl. botulinum type B from the nonproteolytic Cl. botulinum types E and F. The total lipid extracted accounted for 3.7%, 3.3%, 2.0%, 2.7%, and 3.0% of the dry weight of Cl. botulinum vegetative cell types 61E and F; and spore types 61E, F, and 115B, respectively. The fatty acids were analyzed in the form of their methyl esters by gas-liquid chromatography. Infrared spectroscopy, mercuric acetate fractionation, and silver nitrate-thin layer chromatography served as complementary means of analysis. The total fatty acids included straight chain saturated, unsaturated, and cyclopropane acids. Palmitic and myristic were the predominant acids in both the spores and vegetative cells of types 61E, F, and 115B. Together, they made up over 50% of the total fatty acids. Unsaturated acids were the second major group. These were primarily 7, 8-tetradecenoic, 9, 10-hexadecenoic, 7, 8-hexadecenoic, 11, 12-octadecenoic, and 9, 10-octadecenoic acids, Types E and F possessed an 18-carbon diunsaturate, which was not found in the vegetative cells or spores of type 115B. However, insufficient quantities prevented its further characterization. The vegetative cells and spores also contained C₁₅, C₁₇, C₁₉ cyclopropane fatty acids as adjudged by their infrared spectra and gas-liquid chromatographic behavior. The phospholipids accounted for approximately 60% of the total lipids in the vegetative cells and 40% of that in the spores. The primary phospholipid was phosphatidylethanolamine. Qualitative tests for plasmalogens, glycolipids, and phosphatidic acid were positive for both spores and vegetative cells.
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