Temperature and NaCl Affect Growth and Survival of Escherichia coli 0157:H7 in Poultry-Based and Laboratory Media

In tryptic soy broth (TSB) and a poultry extract broth (PB) with 0 to 10% (w/v) NaCl incubated at 37°C, growth of E. coli 0157:H7 was inhibited at 28% NaCl whereas at 10°C, growth was inhibited at 24% NaCl in TSB and at 26% NaCl in PB. The bacterium did not grow at 4°C. Increased NaCl-sensitivity observed at 10°C was a bacteriostatic effect that was ineffective with increasing incubation temperature. At 10°C, E. coli 0157:H7 was more salt-tolerant in PB than in TSB, although PB growth rates were lower. Findings suggest that PB may be a more suitable medium for testing E. coli 0157:H7 in poultry products. Cells of E. coli 0157:H7 that were exposed to refrigeration (4°C) and/or NaCl for 24 days did not grow on MacConkey agar with 1% sorbitol.     

Evaluation of Fourier transform infrared (FT-IR) spectroscopy and chemometrics as a rapid approach for sub-typing Escherichia coli O157:H7 isolates

The importance of tracking outbreaks of foodborne illness and the emergence of new virulent subtypes of foodborne pathogens have created the need for rapid and reliable sub-typing methods for Escherichia coli O157:H7. Fourier transform infrared (FT-IR) spectroscopy coupled with multivariate statistical analyses was used for sub-typing 30 strains of E. coli O157:H7 that had previously been typed by multilocus variable number tandem repeat analysis (MLVA) and pulsed field gel electrophoresis (PFGE). Hierarchical cluster analysis (HCA) and canonical variate analysis (CVA) of the FT-IR spectra resulted in the clustering of the same or similar MLVA types and separation of different MLVA types of E. coli O157:H7. The developed FT-IR method showed better discriminatory power than PFGE in sub-typing E. coli O157:H7. Results also indicated the spectral relatedness between different outbreak strains. However, the grouping of some strains was not in complete agreement with the clustering based on PFGE and MLVA. Additionally, HCA of the spectra differentiated the strains into 30 sub-clusters, indicating the high specificity and suitability of the method for strain level identification. Strains were also classified (97% correct) based on the type of Shiga toxin present using CVA of the spectra. This study demonstrated that FT-IR spectroscopy is suitable for rapid (≤16 h) and economical sub-typing of E. coli O157:H7 with comparable accuracy to MLVA typing. This is the first report of using an FT-IR-based method for sub-typing E. coli O157:H7.     

Effect of environmental stresses on the mean and distribution of individual cell lag times of Escherichia coli O157:H7

The effect of starvation, heat or acid stress on duration of individual cell lag time (τ) and standard deviation (SD) of τ was investigated using Escherichia coli O157:H7. Cells were stressed by exposure to acid (pH 3.5), heat (50 °C), or starvation in either glucose-free mineral medium (MOPS), tryptic soy broth (TSB) or Luria broth (LB). Stressed cells were then diluted into wells of a Bioscreen plate to obtain single cells per well. Replicate time to detection (t_d) values were obtained using the Bioscreen and used to calculate the τ and SD. Significant (P ≤ 0.05) increases in τ over untreated controls were found for the following treatments: 14 days in acid; 2 h of heating; 3 days starvation in MOPS; and 2 days starvation in either TSB or LB. The largest increase in τ was > 2-fold from 2.5 to 5.6 h observed with the heat treatment. MOPS starvation was more detrimental to the cells than was acid treatment over the same time period. A significant increase in SD was found with 21 days acid treatment, and 2 days starvation in either TSB or LB. No significant increase in SD was found for MOPS starvation or heat treatment. Lognormal, Gamma, ExtremeValue and Weibull distributions were fitted to the τ data using BestFit. The results suggest that the Lognormal distribution is suitable for fitting τ data from either stressed or unstressed cells.     

Using of infrared spectroscopy to study the survival and injury of Escherichia coli O157:H7, Campylobacter jejuni and Pseudomonas aeruginosa under cold stress in low nutrient media

The inactivation and sublethal injury of Escherichia coli O157:H7, Campylobacter jejuni and Pseudomonas aeruginosa at three temperatures (22 °C, 4 °C and −18 °C) were studied using traditional microbiological tests and mid-infrared spectroscopy (4000-400 cm⁻¹). Bacteria were cultivated in diluted nutrient matrices with a high initial inoculation (∼10⁷ CFU/ml) levels. Both E. coli O157:H7 and P. aeruginosa survived and cell numbers increased at 22 °C for 5 days while C. jejuni numbers decreased one log₁₀ CFU/ml. A two log CFU/ml decrease was observed for the three pathogens held at 4 °C for 12 days. C. jejuni survived poorly following incubation at −18 °C for 20 days while levels of E. coli O157:H7 and P. aeruginosa remained high (10⁴ CFU/ml). Temperature stress response of microbes was observed by infrared spectroscopy in polysaccharide, protein, lipid, and nucleic acid regions and was strain specific. Level of cold injury could be predicted using cluster, discriminant function and class analog analysis models. Pathogens may produce oligosaccharides and potentially other components in response to stress as indicated by changes in spectral features at 1200-900 cm⁻¹ following freezing.     

Efficacy of various plant hydrosols as natural food sanitizers in reducing Escherichia coli O157:H7 and Salmonella Typhimurium on fresh cut carrots and apples

In the present study, inhibitory effects of the hydrosols of thyme, black cumin, sage, rosemary and bay leaf were investigated against Salmonella Typhimurium and Escherichia coli O157:H7 inoculated to apple and carrots (at the ratio of 5.81 and 5.81 log cfu/g for S. Typhimurium, and 5.90 and 5.70 log cfu/g for E. coli O157:H7 on to apple and carrot, respectively). After the inoculation of S. Typhimurium or E. coli O157:H7, shredded apple and carrot samples were washed with the hydrosols and sterile tap water (as control) for 0, 20, 40 and 60 min. While the sterile tap water was ineffective in reducing (P > 0.05) S. Typhimurium and E. coli O157:H7, 20 min hydrosol treatment caused a significant (P < 0.05) reduction compared to the control group. On the other hand, thyme and rosemary hydrosol treatments for 20 min produced a reduction of 1.42 and 1.33 log cfu/g respectively in the E. coli O157:H7 population on apples. Additional reductions were not always observed with increasing treatment time. Moreover, thyme hydrosol showed the highest antibacterial effect on both S. Typhimurium and E. coli O157:H7 counts. Inhibitory effect of thyme hydrosol on S. Typhimurium was higher than that for E. coli O157:H7. Bay leaf hydrosol treatments for 60 min reduced significantly (P < 0.05) E. coli O157:H7 population on apple and carrot samples. In conclusion, it was shown that plant hydrosols, especially thyme hydrosol, could be used as a convenient sanitizing agent during the washing of fresh-cut fruits and vegetables.