Optimization of broth recovery for repair of heat‐injured Salmonella enterica serovar Typhimurium and Escherichia coli O157:H7

• Published in: Journal of applied microbiology 2019-06, Vol.126 (6), p.1923-1930
• Main Authors: Han, J.‐Y., Song, W.‐J., Kang, D.‐H.
• Format: Article
• Language: English
• Subjects: Bacteria; Brain; broth recovery; Cell suspensions; Diluents; E coli; Escherichia coli; foodborne pathogen; Fruit juices; Heat; heat injury; Heat recovery; injured cell; Injuries; Intervals; Optimization; overlay; Phenols; Repair; Resuscitation; Salmonella; Sorbitol; Yeast
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/jam.14263

Aims The purpose of this research was to determine optimum conditions for broth recovery of heat‐injured Salmonella Typhimurium and Escherichia coli O157:H7. Methods and Results Exposure to 55°C for 15 and 25 min, respectively, induced cellular injury to those pathogens. Comparison was made with the commonly used overlay method using selective medium for recovering sublethally injured cells of S. Typhimurium. For E. coli O157:H7, phenol red agar base with 1% sorbitol was used. After cell suspensions were heated at 55°C for selected time intervals, microbes were 10‐fold diluted with brain heart infusion (BHI), tryptic soy broth (TSB) and TSB with 0·6% yeast extract (TSBYE) and incubated at 37°C for up to 3 h. At hourly intervals, diluents were plated onto selective medium for recovery. Simultaneously, diluents were plated onto tryptic soy agar (TSA) for recovery of sublethally injured cells. For overlays, diluents were plated onto TSA and overlaid with selective agar after a resuscitation interval. Broth recovery conditions for S. Typhimurium and E. coli O157:H7 were determined to be 1 h in any of the following broth media: BHI, TSB or TSBYE. When liquid resuscitation was applied to sublethally injured cells in food samples (milk), 1 h was also sufficient time for recovery. Conclusions The broth recovery method is a convenient alternative to conventional recovery methods. Significance and Impact of the Study Cells sublethally injured by control interventions might not grow on selective medium because they have no resistance to several selective compounds. However, injured cells can recuperate and multiply under conditions sufficient for recovery. To repair and detect heat‐injured cells, the overlay method is commonly used but this method has some limitations. This study confirms the effectiveness of liquid resuscitation method on recovery of injured cells. The broth recovery can replace the overlay method due to greater convenience and timesaving.