Abstract
Grecz, N. (Quartermaster Food and Container Institute, Chicago, Ill.), A. Anellis, and M. D. Schneider. Procedure for cleaning of Clostridium botulinum spores. J. Bacteriol. 84:552–558. 1962.—Liberation of clean spores from vegetative sporangia of Clostridium botulinum strains was accomplished by the use of lytic enzymes and sonic oscillation. Suspensions of crude spores in phosphate buffer (pH 7) were digested with lysozyme (200 μg/ml) and trypsin (100 μg/ml). Rapid lysis of sporangia was induced by ultrasonic oscillation of the reacting mixture at 10 kc for 5 min at 0, 0.5, 1, 2, 4, and 6 hr of incubation at 45 C. Intermittent washing of the reacting spore suspension with a solution of lysozyme and trypsin hastened purification of the spore crop. The cleaning procedure was completed by repeated washing of the spores with distilled water. The spores produced by this procedure were clean, as judged by their microscopic appearance, refractility to staining, loss of heat-sensitive toxin, and partition behavior in a two-phase system composed of polyethylene glycol and 3 m potassium phosphate buffer (pH 7.1). The cleaning procedure appeared not to affect the viability, resistance to heat and gamma radiation, or the toxic nature of C. botulinum spores.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ANELLIS A., KOCH R. B. Comparative resistance of strains of Clostridium botulinum to gamma rays. Appl Microbiol. 1962 Jul;10:326–330. doi: 10.1128/am.10.4.326-330.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BLACK S. H., GERHARDT P. Permeability of bacterial spores. I. Characterization of glucose uptake. J Bacteriol. 1961 Nov;82:743–749. doi: 10.1128/jb.82.5.743-749.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BOYLES W. A., LINCOLN R. E. Separation and concentration of bacterial spores and vegetative cells by foam flotation. Appl Microbiol. 1958 Sep;6(5):327–334. doi: 10.1128/am.6.5.327-334.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BROWN W. L., ORDAL Z. J., HALVORSON H. O. Production and cleaning of spores of putrefactive anaerobe 3679. Appl Microbiol. 1957 May;5(3):156–159. doi: 10.1128/am.5.3.156-159.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CHURCH B. D., HALVORSON H. Dependence of the heat resistance of bacterial endospores on their dipicolinic acid content. Nature. 1959 Jan 10;183(4654):124–125. doi: 10.1038/183124a0. [DOI] [PubMed] [Google Scholar]
- Gaudin A. M., Mular A. L., O'connor R. F. Separation of Microorganisms by Flotation: I. Development and Evaluation of Assay Procedures. Appl Microbiol. 1960 Mar;8(2):84–90. doi: 10.1128/am.8.2.84-90.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gaudin A. M., Mular A. L., O'connor R. F. Separation of Microorganisms by Flotation: II. Flotation of Spores of Bacillus subtilis var. niger. Appl Microbiol. 1960 Mar;8(2):91–97. doi: 10.1128/am.8.2.91-97.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LAWRENCE N. L., HALVORSON H. O. Studies on the spores of aerobic bacteria. IV. A heat resistant catalase from spores of Bacillus terminalis. J Bacteriol. 1954 Sep;68(3):334–337. doi: 10.1128/jb.68.3.334-337.1954. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LONG S. K., WILLIAMS O. B. Method for removal of vegetative cells from Bacterial spore preparations. J Bacteriol. 1958 Sep;76(3):332–332. doi: 10.1128/jb.76.3.332-332.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SACKS L. E., ALDERTON G. Behavior of bacterial spores in aqueous polymer two-phase systems. J Bacteriol. 1961 Sep;82:331–341. doi: 10.1128/jb.82.3.331-341.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
- STEWART B. T., HALVORSON H. O. Studies on the spores of aerobic bacteria. I. The occurrence of alanine racemase. J Bacteriol. 1953 Feb;65(2):160–166. doi: 10.1128/jb.65.2.160-166.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]