Inactivation kinetics of polyphenol oxidase using a two-stage method with low pressurized carbon dioxide microbubbles

Inactivation kinetics of polyphenol oxidase (PPO) by a two-stage method which was additionally pressurized at ambient temperature after carbon dioxide microbubbles (MB-CO₂) was mixed with a solution at low temperature and pressure (two-stage MB-CO₂) was analyzed. The PPO inactivation efficiency of the two-stage MB-CO₂ treatment was higher than that of heat treatment. A decrease in the decimal reduction time (D value) and activation energy during PPO inactivation using the two-stage MB-CO₂ treatment was observed as the temperature of the heating coil and the pressure in the mixing vessel increased. The temperature rise required for a 90% reduction in the D value was increased. Furthermore, the pressure rise required for a 90% reduction in the D value and activation volume increased with the temperature of the heating coil. These results showed that PPO inactivation using the two-stage MB-CO₂ treatment could be achieved with less energy than heat treatment.     

Inactivation of Salmonella enterica serovar Typhimurium and Escherichia coli O157:H7 in peanut butter cracker sandwiches by radio-frequency heating

A multistate outbreak of Salmonella enterica serovar Typhimurium recently occurred in the USA, which was traced back to various food products made with contaminated peanut butter. This study was conducted to investigate the efficacy of radio-frequency (RF) heating to inactivate S. Typhimurium and Escherichia coli O157:H7 in peanut butter cracker sandwiches using creamy and chunky commercial peanut butter and to determine the effect on quality by measuring color changes and sensory evaluation. Samples were treated for a maximum time of 90 s in a 27.12 MHz RF heating system. Samples were prepared in the form of peanut butter cracker sandwiches and placed in the middle of two parallel-plate electrodes. After 90 s of RF treatment, the log reductions of S. Typhimurium and E. coli O157:H7 were 4.29 and 4.39 log CFU/g, respectively, in creamy peanut butter. RF treatment of chunky peanut butter for 90 s also significantly (P < 0.05) reduced levels of S. Typhimurium and E. coli O157:H7 by 4.55 log CFU/g and 5.32 log CFU/g. Color values and sensory characteristics of the RF treated peanut butter and crackers were not significantly (P > 0.05) different from the control. These results suggest that RF heating can be applied to control pathogens in peanut butter products without affecting quality.     

Bovine cathepsin D activity under high pressure

The stability and catalytic activity of bovine cathepsin D in Bis-Tris buffer (pH 6.0) in different pressure–temperature domains (0.1–650 MPa, 20–75 °C) were investigated and described with mathematical models. Cathepsin D inactivation followed first-order kinetics at all pressure–temperature conditions tested. The protease was largely pressure stable at room temperature and heat stable at ambient pressure up to 300 MPa and 55 °C, respectively, causing less than 10% inactivation after 10 min treatment. Pressure and temperature act synergistically on the enzyme inactivation under most conditions. However, at 100 MPa a significant stabilisation of the enzyme against temperature-induced inactivation was observed. Pressure drastically inhibited the cleavage of a synthetic substrate by cathepsin D in Bis-Tris buffer (pH 6.0) causing a reduction of the catalytic rate of more than 50% at 100–400 MPa. Maximal substrate cleavage by cathepsin D was identified at 60 °C and ambient pressure conditions after 20 min treatment.     

Food treatment with high pressure carbon dioxide: Saccharomyces cerevisiae inactivation kinetics expressed as a function of CO2 solubility

Experimental survival curves of Saccharomyces cerevisiae cells exposed to high pressure carbon dioxide (HPCD) treatments under several constant temperatures (35, 40 and 50 °C), pressures (7.5, 10.0 and 13.0 MPa) and suspended in distilled water with different sodium phosphate monobasic buffer concentrations (0.02, 0.10, 0.20 and 0.40 M) were obtained. The Peleg model was applied to the isobaric and isothermal conditions described by the power law equation log[S(t)] = −btⁿ, where S(t) is the momentary survival ratio and ‘b’ and ‘n’ are the rate and the shape parameters, respectively. The values of the coefficients ‘b’ and ‘n’ were calculated for each experiment at fixed pressure and temperature. For each suspending medium the power law model was proposed to describe the combined effects of pressure and temperature. Taking into account the CO₂ solubility as a function of the sodium phosphate monobasic concentration, ‘b’ and ‘n’ were correlated to the CO₂ solubility values and temperature. An equation was proposed for ‘b’ as a function of CO₂ solubility and temperature while ‘n’ was a weak function of temperature. The resulting equation was much simpler that the one obtained correlating the microbial inactivation to pressure and temperature and, more important, it was independent of the suspending medium. The results indicate that the coupling of carbon dioxide solubility, also predicted with commercial software, and the use of inactivation models referred to solubility and temperature may provide a powerful instrument for the interpretation of microbial inactivation experiments and for the design of HPCD processes and equipments.     

Inactivation of Escherichia coli O157:H7 during thermophilic anaerobic digestion of manure from dairy cattle

Inactivation of the pathogenic Escherichia coli serotype O157:H7 and a non-pathogenic E. coli strain isolated from dairy cattle manure was evaluated with batch tests at 50 and 55 degrees C in biosolids from a thermophilic anaerobic digester treating the manure. Using differential-selective plating on sorbitol-MacConkey (SMAC) agar to quantify E. coli, the decline in concentrations of both the sorbitol-negative (putative E. coli O157:H7) and sorbitol-positive (putative non-pathogenic E. coli) organisms followed a model that assumed there was a heat-sensitive fraction and a heat-resistant fraction. Inactivation rates of the heat-sensitive fractions were similar for both colony types at each temperature, suggesting that wild-type E. coli can be used as an indicator of inactivation of serotype O157:H7. The decimal reduction time for the heat-sensitive fractions was in the order of 10min at 55 degrees C and ranged from approximately 1-3h at 50 degrees C. Concentrations of heat-resistant organisms at 55 degrees C were 1.4-1.7log(10)cfu/mL. Confirmatory analyses conducted on 30 randomly selected colonies of heat-resistant sorbitol-negative cells from treatment at 55 degrees C indicated that none were serotype O157:H7, nor were they E. coli. Similar analyses on 10 sorbitol-negative isolates from untreated manure indicated that none were serotype O157:H7, although 16S rRNA gene sequence analysis indicated that eight were E. coli or closely related enteric bacteria. These findings suggest that plating on differential-selective media to quantify E. coli, including serotype O157:H7, in effluent samples from thermophilic anaerobic digestion can lead to false positive results. Therefore, more specific methods should be used to evaluate the extent of thermal inactivation of both pathogenic and non-pathogenic E. coli in manure treatment systems.     

Mechanisms of Escherichia coli inactivation by several disinfectants

The objective of this study was to elucidate dominant mechanisms of inactivation, i.e. surface attack versus intracellular attack, during application of common water disinfectants such as ozone, chlorine dioxide, free chlorine and UV irradiation. Escherichia coli was used as a representative microorganism. During cell inactivation, protein release, lipid peroxidation, cell permeability change, damage in intracellular enzyme and morphological change were comparatively examined. For the same level of cell inactivation by chemical disinfectants, cell surface damage was more pronounced with strong oxidant such as ozone while damage in inner cell components was more apparent with weaker oxidant such as free chlorine. Chlorine dioxide showed the inactivation mechanism between these two disinfectants. The results suggest that the mechanism of cell inactivation is primarily related to the reactivity of chemical disinfectant. In contrast to chemical disinfectants, cell inactivation by UV occurred without any changes measurable with the methods employed. Understanding the differences in inactivation mechanisms presented herein is critical to identify rate-limiting steps involved in the inactivation process as well as to develop more effective disinfection strategies.     

Inactivation of peroxidase and polyphenol oxidase in red beet (Beta vulgaris L.) extract with continuous high pressure carbon dioxide

The inactivation of peroxidase (POD) and polyphenol oxidase (PPO) in red beet extract (RBE) with continuous high pressure carbon dioxide (HPCD) was investigated. HPCD treatment at 7.5 MPa (55 °C, 30 min) resulted in a reduction of their activities by approximately 73% and 93%, respectively. Compared with thermal treatment, continuous HPCD treatment reduced the decimal reduction time (D) of POD and PPO from 555.6 min to 55.9 min and 161.3 min to 32.1 min, respectively. The inactivation process could be described by first-order kinetics (r² > 0.70, p < 0.05); D values declined when temperature increased and continuous HPCD at 7.5 MPa and 55 °C resulted in the highest reaction rate constant (k value; smallest D value). The activation energy of the inactivation was reduced by HPCD treatment from 92.5 kJ/mol to 69.8 kJ/mol and 57.1 kJ/mol to 49.5 kJ/mol for POD and PPO, respectively. Continuous HPCD treatment had little effect on the antioxidant capacities of RBE samples.     

Electrochemical inactivation of coliforms by in-situ generated hydroxyl radicals

Electrochemical disinfection is quite attractive as a promising alternative technology to chlorination. It is still debated whether conventional electrochemical disinfection, which electrolyzes the solution with very high chloride concentration to produce excess amounts of chlorine species, will generate toxic disinfection byproducts (DBPs) and have the same health risks as chlorination. To resolve this critical issue, we explored the possibility of electrochemical disinfection based on electrogenerated free radicals but not on active chlorine. The germicidal efficiency of 99.99% was achieved with a contact time of 5 min and current density of 7 mA cm⁻² for a chloride-free model wastewater contaminated by coliforms. Electron spin resonance detection clearly confirmed that hydroxyl radicals were the major germicidal species responsible for efficient electrochemical disinfection. This process would not generate poisonous DBPs due to the avoidance of dangerous chlorine species. pH in the range of 5–9 has little effect on the bacteria inactivation. Formation mechanism of hydroxyl radicals was discussed.     

Nature of the inactivation of Escherichia coli suspended in an orange juice and milk beverage

The killing effect of pulsed electric fields (PEF) on Escherichia coli (ATCC 8739) suspended in an orange juice and milk beverage was studied. Bipolar square pulses with a pulse width of 2.5 μs were applied. Electric field strength and treatment times ranged from 15 to 40 kV/cm, and from 0 to 700 μs, respectively. A maximum of 3.83 log reductions was achieved at 15 kV/cm and 700 μs. The experimental data were fitted to Bigelow and Hülsheger models and Weibull distribution function. Results indicated that Weibull function best described the experimental data (lowest mean square error). As there were no significant differences in the values of the shape factor (n) at the electric field strength of 25–40 kV/cm, the number of parameters in the Weibull model were reduced, leading to a simplified model with a fit similar to that obtained with the full model.     

Destruction of Helminth (Ascaris suum) Eggs by Ozone

Based on the properties of ozone as a strong germicidal agent, the inactivation kinetics or disintegration of Ascaris suum eggs in water were studied. Ozone was applied in synthetic samples, the concentration of dissolved ozone in the liquid phase was typically between 3.5 to 4.7mg/L, pH 5, 9. The value of 90% inactivation (t₉₀) obtained was at 1 h, the remaining 10% being inactivated in 2 h. A linear correlation between the logarithm of bacterial or yeast concentration (N) and contact time was found, being the linear significant correlation coefficient (r of 0.95). That 2-log inactivation occurred at a CT value near 4.7mg/min/L. The best reduction percentage was of 99.00 in 120min. It therefore could be demonstrated that ozone indeed induces effects on the helminth eggs.