Influence of tonicity and chloramphenicol on hyperthermic cytotoxicity and cell permeability under various heating rates

Abstract

The cell lethality and permeability induced in Escherichia coli E/r, Escherichia coli B_s-1 and Zygosaccharornyces bailii cells by high temperature (52°C) after heating at different rates (mean s 0.015, 0.25 and 150°C per s) and in media of different tonicity and content (isotonic YEP broth versus 0.01 M phosphate buffer, pH7.0 containing different concentrations of NaCl) and with versus without chloramphenicol (10pg/ml) have been investigated. Hyperthermic treatment in YEP broth or isotonic 0.01 M phosphate buffer resulted in markedly reduced cytotoxicity with decreasing heat rate. The heating rate effect was larger when the cells were treated in YEP broth. Chloramphenicol, which is known to inhibit expression of heat shock proteins in bacteria, did not affect the viability of cells or the development of thermotolerance in cells heated at different heating rates in isotonic phosphate buffer but prevented the development of an additional degree of thermotolerance in cells heated slowly in YEP broth. In contrast, the differential effect of heating rate on cytotoxicity and cell permeability was not demonstrated when cells were heated in hypertonic solution (1 M NaCl in 0.01 M phosphate buffer, pH 7.0). It is proposed that heat destabilization of the osmotic cell homeostasis, which is more profound after rapid heating, plays a major part in heat induced cellular lethality.

Key Words:

• Heating rate
• viability
• permeability
• osmotic cell homeostasis
• tonicity