Strain and serovar variants of Salmonella enterica exhibit diverse tolerance to food chain-related stress.

Non-Typhoidal Salmonella (NTS) continues to be a leading cause of foodborne illness worldwide. Food manufacturers implement hurdle technology by combining more than one approach to control food safety and quality, including preservatives such as organic acids, refrigeration, and heating. We assessed the variation in survival in stresses of genotypically diverse isolates of Salmonella enterica to identify genotypes with potential elevated risk to sub-optimal processing or cooking. Sub-lethal heat treatment, survival in desiccated conditions and growth in the presence of NaCl or organic acids were investigated. S. Gallinarum strain 287/91 was most sensitive to all stress conditions. While none of the strains replicated in a food matrix at 4 °C, S. Infantis strain S1326/28 retained the greatest viability, and six strains exhibited a significantly reduced viability. A S. Kedougou strain exhibited the greatest resistance to incubation at 60 °C in a food matrix that was significantly greater than S. Typhimurium U288, S Heidelberg, S. Kentucky, S. Schwarzengrund and S. Gallinarum strains. Two isolates of monophasic S. Typhimurium, S04698-09 and B54Col9 exhibited the greatest tolerance to desiccation that was significantly more than for the S. Kentucky and S. Typhimurium U288 strains. In general, the presence of 12 mM acetic acid or 14 mM citric acid resulted in a similar pattern of decreased growth in broth, but this was not observed for S. Enteritidis, and S. Typhimurium strains ST4/74 and U288 S01960-05. Acetic acid had a moderately greater effect on growth despite the lower concentration tested. A similar pattern of decreased growth was observed in the presence of 6% NaCl, with the notable exception that S. Typhimurium strain U288 S01960-05 exhibited enhanced growth in elevated NaCl concentrations.

Cleaning and sanitation of Salmonella-contaminated peanut butter processing equipment.

Microbial contamination of peanut butter by Salmonella poses a significant health risk as Salmonella may remain viable throughout the product shelf life. Effective cleaning and sanitation of processing lines are essential for preventing cross-contamination. The objective of this study was to evaluate the efficacy of a cleaning and sanitation procedure involving hot oil and 60% isopropanol, ± quaternary ammonium compounds, to decontaminate pilot-scale processing equipment harboring Salmonella. Peanut butter inoculated with a cocktail of four Salmonella serovars (∼ 7 log CFU/g) was used to contaminate the equipment (∼ 75 L). The system was then emptied of peanut butter and treated with hot oil (90 °C) for 2 h followed by sanitizer for 1 h. Microbial analysis of food-contact surfaces (7 locations), peanut butter, and oil were conducted. Oil contained ∼ 3.2 log CFU/mL on both trypticase soy agar with yeast extract (TSAYE) and xylose lysine deoxycholate (XLD), indicating hot oil alone was not sufficient to inactivate Salmonella. Environmental sampling found 0.25-1.12 log CFU/cm(2) remaining on processing equipment. After the isopropanol sanitation (± quaternary ammonium compounds), no Salmonella was detected in environmental samples on XLD (<0.16 log CFU/cm(2)). These data suggest that a two-step hot oil clean and isopropanol sanitization treatment may eliminate pathogenic Salmonella from contaminated equipment.

Comparison of Antibacterial Activity of Phytochemicals against Common Foodborne Pathogens and Potential for Selection of Resistance.

Antimicrobial resistance is now commonly observed in bacterial isolates from multiple settings, compromising the efficacy of current antimicrobial agents. Therefore, there is an urgent requirement for efficacious novel antimicrobials to be used as therapeutics, prophylactically or as preservatives. One promising source of novel antimicrobial chemicals is phytochemicals, which are secondary metabolites produced by plants for numerous purposes, including antimicrobial defence. In this report, we compare the bioactivity of a range of phytochemical compounds, testing their ability to directly inhibit growth or to potentiate other antimicrobials against Salmonella enterica Typhimurium, Pseudomonas aeruginosa, Listeria monocytogenes, and Staphylococcus aureus. We found that nine compounds displayed consistent bioactivity either as direct antimicrobials or as potentiators. Thymol at 0.5 mg/mL showed the greatest antimicrobial effect and significantly reduced the growth of all species, reducing viable cell populations by 66.8%, 43.2%, 29.5%, and 70.2% against S. enterica Typhimurium, S. aureus, P. aeruginosa, and L. monocytogenes, respectively. Selection of mutants with decreased susceptibility to thymol was possible for three of the pathogens, at a calculated rate of 3.77 × 10-8, and characterisation of S. enterica Typhimurium mutants showed a low-level MDR phenotype due to over-expression of the major efflux system AcrAB-TolC. These data show that phytochemicals can have strong antimicrobial activity, but emergence of resistance should be evaluated in any further development.

Development of a high pressure processing inactivation model for hepatitis A virus.

High pressure processing (HPP) inactivation data were obtained for hepatitis A virus (HAV) suspended in buffered growth medium containing salt at either 15 or 30 g/liter. Pressures between 300 and 500 MPa were applied for treatment times of 60 to 600 s. In medium containing 15 g/liter salt, the HAV titer was reduced by approximately 1 and 2 log 50% tissue culture infectious dose units (TCID50) per ml after 600 s of treatment with 300 and 400 MPa, respectively. Under the same treatment conditions but in medium containing 30 g/liter salt, HAV was reduced by <0.50 log TCID50/ml. HAV was inactivated by >3 log TCID50/ml after treatment with 500 MPa for 300 and 360 s in medium containing 15 and 30 g/liter salt, respectively. Weibull and log-linear models were fitted to inactivation data. Individual Weibull curves generally provided a good fit at each pressure and salinity, but the curve shapes were qualitatively inconsistent between treatments, making interpolation between pressures difficult and unreliable. High variability was observed in the inactivation data, but the log-linear model described the entire data set and interpolated between specific treatment conditions. Therefore, this model was evaluated by using high pressure to treat HAV artificially inoculated into Pacific oyster (Crassostrea gigas) homogenate adjusted to 15 or 30 g/liter salinity. The log-linear model generally provided fail-safe predictions at pressures > or = 375 MPa and may aid shellfish processors wishing to incorporate HPP into an oyster processing regime. Additional inactivation data with greater reproducibility should be collected to enable expansion of the model and to increase the accuracy of its predictions.

Inactivation of foodborne viruses of significance by high pressure and other processes.

The overall safety of a food product is an important component in the mix of considerations for processing, distribution, and sale. With constant commercial demand for superior food products to sustain consumer interest, nonthermal processing technologies have drawn considerable attention for their ability to assist development of new products with improved quality attributes for the marketplace. This review focuses primarily on the nonthermal processing technology high-pressure processing (HPP) and examines current status of its use in the control and elimination of pathogenic human viruses in food products. There is particular emphasis on noroviruses and hepatitis A virus with regard to the consumption of raw oysters, because noroviruses and hepatitis A virus are the two predominant types of viruses that cause foodborne illness. Also, application of HPP to whole-shell oysters carries multiple benefits that increase the popularity of HPP usage for these foods. Viruses have demonstrated a wide range of sensitivities in response to high hydrostatic pressure. Viral inactivation by pressure has not always been predictable based on nomenclature and morphology of the virus. Studies have been complicated in part from the inherent difficulties of working with human infectious viruses. Consequently, continued study of viral inactivation by HPP is warranted.

Norovirus cross-contamination during preparation of fresh produce.

Infection with human norovirus (HuNoV) is considered a common cause of foodborne illness worldwide. Foodborne HuNoV outbreaks may result from consumption of food contaminated by an infected food handler in the foodservice environment, in which bare-hand contact, lack of hand washing, and inadequate cleaning and disinfection are common contributing factors. The goal of this study was to examine cross-contamination of a HuNoV surrogate, murine norovirus (MNV-1), during common procedures used in preparing fresh produce in a food service setting, including turning water spigots, handling and chopping Romaine lettuce, and washing hands. MNV-1 transfer % was log-transformed to achieve a normal distribution of the data and enable appropriate statistical analyses to be performed. MNV-1 transfer coefficients varied by surface type, and a greater affinity for human hands and chopped lettuce was observed. For example, greater transfer was observed from a contaminated stainless steel spigot to a clean hand (24% or 1.4-log transfer %) compared to transfer from hand to spigot (0.6% or -0.2-log transfer %). During the chopping of Romaine lettuce, MNV-1 was transferred from either a contaminated cutting board (25% or 1.4-log transfer %) or knife (~100% or 2.0-log transfer %) to lettuce at a significantly greater rate (p>0.05) than from contaminated lettuce to the board (2.1% or 0.3-log transfer %) and knife (1.2% or 0.06-log transfer %). No significant difference (p>0.05) in MNV-1 transfer coefficients was observed between bare hands and Romaine lettuce during handling. For handwashing trials, only one hand was inoculated with MNV-1 prior to washing. The handwashing methods included rubbing hands under tap water for at least 5s (average 2.8-log reduction) or washing hands for at least 20s with liquid soap (average 2.9-log reduction) or foaming soap (average 3.0-log reduction), but no statistical difference between these reductions was achieved (p>0.05). Despite the reductions of MNV-1 observed, residual virions were detected on both hands after washing in every replicate trial. This observation reveals that virions are transferred from one hand to the other during washing with and without soap. Each transfer scenario was repeated at least 9 times, and the data gathered indicate that MNV-1 transfers readily between common surfaces during food preparation. These data are important for the development of quantitative risk analyses, and will assist in the development of appropriate intervention strategies for enteric viruses in food preparation.

Effect of growth on the thermal resistance and survival of Salmonella Tennessee and Oranienburg in peanut butter, measured by a new thin-layer thermal death time device.

In published data the thermal destruction of Salmonella species in peanut butter deviates from pseudo-first-order kinetics. The reasons for such deviation are unknown. This study examined both the method used to measure the thermal destruction rate and the method of growth of the microorganisms to explain variations in destruction kinetics. Growth on a solid matrix results in a different physiological state that may provide greater resistance to adverse environments. In this study, Salmonella Tennessee and Oranienburg were grown for 24 h at 37°C under aerobic conditions in broth and agar media to represent planktonic and sessile cell growth, respectively. Peanut butter was held at 25°C and tested for Salmonella levels immediately after inoculation and at various time intervals up to 2 weeks. Thermal resistance was measured at 85°C by use of a newly developed thin-layer metal sample holder. Although thermal heat transfer through the metal device resulted in longer tau values than those obtained with plastic bags (32.5 ± 0.9 versus 12.4 ± 1.9 s), the bags have a relative variability of about 15 % compared with about 3 % in the plates, allowing improved uniformity of sample treatment. The two serovars tested in the thin-layer device showed similar overall thermal resistance levels in peanut butter regardless of growth in sessile or planktonic states. However, thermal destruction curves from sessile cultures exhibited greater linearity than those obtained from planktonic cells (P = 0.0198 and 0.0047 for Salmonella Oranienburg and Salmonella Tennessee, respectively). In addition, both Salmonella serovars showed significantly higher survival in peanut butter at 25°C when originally grown on solid media (P = 0.001) with a <1-log loss over 2 weeks as opposed to a 1- to 2-log loss when grown in liquid culture. Consequently, the use of cells grown on solid media may more accurately assess the survival of Salmonella at different temperatures in a low-water-activity environment such as peanut butter.