Heat Resistance, Virulence, and Gene Expression of Desiccation-Adapted Salmonella Enteritidis During Long-Term Storage in Low-Water Activity Foods.

Desiccation stress could induce crossprotection and even affect virulence of Salmonella enterica. However, the influence of food matrices with low-water activity on desiccation adaptation of Salmonella still remains unclear. This study investigated the survival and adaptation of Salmonella Enteritidis in skim milk powder, ginger powder, and chocolate powder under desiccation storage conditions for a total of 12 weeks. High survival rates of Salmonella Enteritidis in all food matrices maintained over the long-term desiccation storage. Desiccation-adapted Salmonella Enteritidis enhanced heat resistance (p < 0.05) with the increase of storage time. Food composition plays an important role in the induction of crossresistance of desiccation-adapted Salmonella. After desiccation storage, Salmonella Enteritidis in ginger powder was most tolerant to heat treatment. Salmonella Enteritidis in skim milk powder was most resistant to the gastrointestinal simulation environment, and had strongest adhesion to Caco-2 cells. The effects of food composition on gene expression (rpoS, proV, otsA, otsB, grpE, dnaK, rpoH, and sigDE) in desiccation-adapted Salmonella Enteritidis were not significant (p > 0.05). At initial desiccation storage, osmotic protection-related genes (fadA, proV, otsA, and otsB), stress response regulator (rpoS), and heat-resistance-related genes (grpE, dnaK, and rpoH) were all significantly upregulated (p < 0.05). However, after 4-week storage, the expression level of desiccation-related genes, proV, otsA, otsB, grpE, dnaK, and rpoH, significantly decreased (p < 0.05). This study enables a better understanding of Salmonella's responses to long-term desiccation stress in different kinds of low-water activity foods.

Induction of a Viable but Nonculturable State, Thermal and Sanitizer Tolerance, and Gene Expression Correlation with Desiccation-Adapted Biofilm and Planktonic Salmonella in Powdered Infant Formula.

ABSTRACT: This study was conducted to investigate the effects of the physiological state, desiccation adaptation, and storage of powdered infant formula on Salmonella cell survival and expression of desiccation stress-related genes. Powdered infant formula was inoculated with Salmonella Typhimurium in the biofilm state on beads and in the planktonic state on nitrocellulose filters and stored at 25°C for up to 270 days. Both 5-cyano-2,3-ditoyl tetrazolium chloride flow cytometry and xylose lysine deoxycholate agar plate counts revealed that biofilm-forming Salmonella cells tended to enter the viable but nonculturable (VBNC) state (P < 0.05). The population reduction of all desiccation-adapted Salmonella Typhimurium decreased significantly in both physiological states after exposure to mild heat (60°C) compared with nonadapted control cells (P < 0.05). Salmonella cells were cross-protected from heat in both physiological states, but cross-protection against hydrogen peroxide was induced for only planktonic Salmonella cells. The reverse transcription quantitative PCR results revealed that the rpoS gene in biofilm Salmonella cells on all desiccation adaption days and in planktonic Salmonella cells on day 7 of dry storage was significantly upregulated (P < 0.05). The rpoE, grpE, and invA genes in Salmonella cells in both physiological states were significantly down-regulated (P < 0.05). Physiological state and storage time might affect expression of these genes. Prior exposure to adverse conditions, including low water activity, and the physiological state impacted Salmonella survival, and its ability to enter the VBNC state and gene expression.