Synergistic Effects of Nisin, Lysozyme, Lactic Acid, and CitricidalTM for Enhancing Pressure-Based Inactivation of Bacillus amyloliquefaciens, Geobacillus stearothermophilus, and Bacillus atrophaeus Endospores

The inactivation of bacterial endospores continues to be the main curtailment for further adoption of high-pressure processing in intrastate, interstate, and global food commerce. The current study investigated the effects of elevated hydrostatic pressure for the inactivation of endospore suspension of three indicator spore-forming bacteria of concern to the food industry. Additionally, the effects of four bacteriocin/bactericidal compounds were studied for augmenting the decontamination efficacy of the treatment. Elevated hydrostatic pressure at 650 MPa and at 50 °C was applied for 0 min (untreated control) and for 3, 7, and 11 min with and without 50K IU of nisin, 224 mg/L lysozyme, 1% lactic acid, and 1% CitricidalTM. The results were statistically analyzed using Tukey- and Dunnett’s-adjusted ANOVA. Under the condition of our experiments, we observed that a well-designed pressure treatment synergized with mild heat and bacteriocin/bactericidal compounds could reduce up to >4 logs CFU/mL (i.e., >99.99%) of bacterial endospores. Additions of nisin and lysozyme were able, to a great extent, to augment (p < 0.05) the decontamination efficacy of pressure-based treatments against Bacillus amyloliquefaciens and Bacillus atrophaeus, while exhibiting no added benefit (p ≥ 0.05) for reducing endospores of Geobacillus stearothermophilus. The addition of lactic acid, however, was efficacious for augmenting the pressure-based reduction of bacterial endospores of the three microorganisms.

Sensitivity of wild-type and rifampicin-resistant O157 and non-O157 Shiga toxin-producing Escherichia coli to elevated hydrostatic pressure and lactic acid in ground meat and meat homogenate

Various serogroups of Shiga toxin-producing Escherichia coli have been epidemiologically associated with foodborne disease episodes in the United States and around the globe, with E. coli O157: H7 as the dominant serogroup of public health concern. Serogroups other than O157 are currently associated with about 60% of Shiga toxin-producing E. coli related foodborne illness episodes. Current study evaluated sensitivity of the O157 and epidemiologically important non-O157 serogroups of the pathogen to elevated hydrostatic pressure and 1% lactic acid. Pressure intensity of 250 to 650 MPa were applied for 0 to 7 min for inactivation of strain mixtures of wild-type and rifampicin-resistant E. coli O157, as well as O26, O45, O103, O111, O121, and O145 serogroups and ATCC® 43895™ strain in ground meat and 10% meat homogenate. E. coli O157 were reduced (p < 0.05) from 6.86 ± 0.2 to 4.56 ± 0.1 log CFU/g when exposed to pressure of 650 MPa for 7 min. Corresponding reductions (p < 0.05) for non-O157 E. coli were from 6.98 ± 0.3 to 4.72 ± 0.1. The D-values at 650 MPa were 3.71 and 3.47 min for O157 and non-O157 serogroups, respectively. Presence of 1% lactic acid to a great extent augmented (p < 0.05) decontamination efficacy of the treatment in meat homogenate resulting in up to 5.6 and 6.0 log CFU/mL reductions for O157 and non-O157 serogroups, respectively. Among the tested serogroups, the wild-type and rifampicin-resistant phenotypes exhibited (p ≥ 0.05) comparable pressure sensitivity. Thus, these two phenotypes could be used interchangeably in validation studies. Our results also illustrate that, application of elevated hydrostatic pressure could be utilized for assuring safety of ground and non-intact meat products against various serogroups of Shiga toxin-producing E. coli. Addition of 1% lactic acid additionally provided industrially appreciable augmentation in efficacy of the pressure-based treatments.

Sensitivity of Planktonic Cells of Staphylococcus aureus to Elevated Hydrostatic Pressure as Affected by Mild Heat, Carvacrol, Nisin, and Caprylic Acid

Current study investigated effects of elevated hydrostatic pressure exposure in the presence of mild heat and natural antimicrobials against Staphylococcus aureus. Hydrostatic pressure of 350 to 550 MPa with nisin (5000 IU/mL), carvacrol, or caprylic acid (0.5% v/v) were applied for the reduction in four-strain mixture of S. aureus in HEPES buffer at 4 and 40 °C for up to 7 min. Results were statistically analyzed by ANOVA and D-values were additionally calculated using best-fitted linear model. Prior to exposure to treatments at 4 °C, counts of the pathogen were 7.95 ± 0.4 log CFU/mL and were reduced (p < 0.05) to 6.44 ± 0.3 log CFU/mL after 7 min of treatment at 450 MPa. D-value associated with this treatment was 5.34 min (R2 = 0.72). At 40 °C, counts were 8.21 ± 0.7 and 5.77 ± 0.3 log CFU/mL before and after the 7-min treatments, respectively. D-value associated with 40 °C treatment was 3.30 min (R2 = 0.62). Application of the antimicrobials provided additional pathogen reduction augmentation for treatments < 5 min. The results of the current study could be incorporated for meeting regulatory requirements such as Food Code, HACCP, and Preventive Control for Human Food of Food Safety Modernization Act for assuring microbiological safety of products against this prevalent pathogen of public health concern.

Augmenting the Pressure-Based Pasteurization of Listeria monocytogenes by Synergism with Nisin and Mild Heat

The current study investigated Listeria monocytogenes inactivation using mild heat with elevated hydrostatic pressure and nisin under buffered condition. A four-strain pathogen mixture was exposed to 0 (control) and up to 9 min of (1) 4 °C elevated pressure; (2) 4 °C elevated pressure and nisin; (3) 4 °C nisin; (4) heat at 40 °C; (5) 40 °C elevated pressure; (6) 40 °C elevated pressure and nisin; and (7) 40 °C nisin. Elevated hydrostatic pressure at 400 MPa (Hub880 Explorer, Pressure BioScience Inc., Easton, MA, USA) and nisin concentration of 5000 IU/mL were used in the trials. Analyses of variance were conducted, followed by Dunnett’s- and Tukey-adjusted means separations. Under conditions of these experiments, nisin augmented (p < 0.05) decontamination efficacy of 40 °C heat and elevated hydrostatic pressure treatments, particularly at treatment interval of 3 min. This synergism with nisin faded away (p ≥ 0.05) as the treatment time for thermal, high-pressure, and thermal-assisted pressure processing increased. The results of our study, thus, exhibit that practitioners and stakeholders of pressure-based technologies could benefit from synergism of mild heat and nisin for short-term, high-pressure pasteurization treatments to achieve microbial safety and economic feasibility comparable to traditional heat-treated products.