This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Iannetta, A. Articles by Wedgwood, R. J. Search for Related Content PubMed PubMed Citation Articles by Iannetta, A. Articles by Wedgwood, R. J.

Culture of Serum-induced Spheroplasts from Vibrio cholerae

¹ Department of Pediatrics, University of Washington, School of Medicine, Seattle, Washington 98105

ABSTRACT

Spheroplasts of Vibrio cholerae types Inaba, Hikojima, and Ogawa were produced in serum. An excess of lysozyme was added to expedite transformation, and CaCl₂ was used as the stabilizing agent. At the optimal time for each strain, when less than 1% classical rod forms were observed microscopically, samples were plated on both conventional and modified L-form media. No growth occurred on conventional media; L-form type colonies appeared on L-form media after 2 to 6 days of incubation. L-form colonies could be subcultured on conventional media and required from four to six passages for complete reversion to classical parent forms. Reaction mixtures of Hikojima and Inaba types were passed through membrane filters (0.45 µ); L-form colonies were grown from both strains after spheroplast transformation. Appropriate controls were negative. It is suggested that the replicative particle may be filtrable.