Effect of cell wall treatment using high pressure on CARD-FISH method

Abstract

 We investigated the effect of cell wall treatment utilizing high pressure on the CARD-FISH method against Escherichia coli and Rhodococcus equi cells as a Gram-negative and Gram-positive bacteria respectively. As a result, fluorescence signals from both E. coli and R. equi cells could not be confirmed under almost all conditions with high pressure treatment alone. However, it was revealed that by combining high pressure treatment with enzyme treatment such as lysozyme and achromopeptidase, a higher fluorescence signal can be obtained than each treatment alone. It is possible to obtain a strong fluorescent signal even for microorganisms with different cell wall structures by carrying out cell wall treatment with appropriate conditions such as pressure level, treatment temperature, enzyme concentration. It was shown that high pressure treatment can be applied as a versatile and effective cell wall treatment in the CARD-FISH method.