Separation of active and inactive fractions from starved culture of Vibrio parahaemolyticus by density dependent cell sorting

• Published in: FEMS microbiology ecology Vol. 51; no. 2; pp. 179 - 186
• Main Authors: Nayak, Binaya Bhusan, Kamiya, Eriko, Nishino, Tomohiko, Wada, Minoru, Nishimura, Masahiko, Kogure, Kazuhiro
• Format: Journal Article
• Language: English
• Published: Oxford, UK Elsevier B.V 2005; Blackwell Publishing Ltd; Blackwell; Oxford University Press
• Subjects: Bacteriology; Biological and medical sciences; Cell culture; Cell Respiration; Cell Separation - methods; Cell specific activity; Centrifugation, Density Gradient - methods; Density; Density dependent cell sorting; Ecology; Fundamental and applied biological sciences. Psychology; Leucine; Microbiology; Miscellaneous; Percoll gradient; Reduction; Viable but nonculturable; Vibrio parahaemolyticus; Vibrio parahaemolyticus - cytology; Vibrio parahaemolyticus - isolation & purification; Vibrio parahaemolyticus - metabolism; Waterborne diseases; Density dependent cell sorting; Viable but nonculturable; Vibrio parahaemolyticus; Cell specific activity; Percoll gradient; Cell culture; Gradient; Microbiology; Ecology; Leucine; Method; Cell fraction; Bacteria; Vibrionaceae
• ISSN: 0168-6496; 1574-6941
• DOI: 10.1016/j.femsec.2004.08.005

The co-existence of physiologically different cells in bacterial cultures is a general phenomenon. We have examined the applicability of the density dependent cell sorting (DDCS) method to separate subpopulations from a long-term starvation culture of Vibrio parahaemolyticus. The cells were subjected to Percoll density gradient and separated into 12 fractions of different buoyant densities, followed by measuring the cell numbers, culturability, respiratory activity and leucine incorporation activity. While more than 78% of cells were in lighter fractions, about 95% of culturable cells were present in heavier fractions. The high-density subpopulations also had high proportion of cells capable of forming formazan granules. Although this was accompanied by the cell specific INT-reduction rate, both leucine incorporation rates and INT-reduction rates per cell had a peak at mid-density fraction. The present results indicated that DDCS could be used to separate subpopulations of different physiological conditions.