Induction of a viable but non-culturable state in Salmonella Typhimurium by thermosonication and factors affecting resuscitation

The objective of this work was to analyze the effects of thermosonication (TS) on induction of a viable but non-culturable (VBNC) state in Salmonella Typhimurium and to examine incubation factors affecting subsequent resuscitation of cells. A TS treatment of 380 W at 53°C for 30 min induced the VBNC state in S. Typhimurium cells in beef peptone yeast (BPY) broth, apple/carrot juice, physiological saline and phosphate buffer solution. The logarithmic and decline phases of growth were more sensitive to the TS treatment compared to stationary phase cells. Meanwhile, VBNC S. Typhimurium could be resuscitated back to culturable cells by using suitable incubation temperatures and media. Addition of Tween 20 hindered resuscitation compared to the use of BPY medium alone. The optimal growth temperature (i.e. 37°C) was the most suitable temperature to resuscitate cells from the VBNC state. The VBNC incidence index decreased with the addition of sodium pyruvate during TS treatment, as it accelerated resuscitation. The results demonstrated that free radicals produced during TS processing and the growth phase of cells affected induction of the VBNC state in S. Typhimurium. Hence, the kinds and amounts of free radicals generated during TS treatment should be analyzed in the future.