Theoretical and Experimental Investigation of the Thermal Inactivation of Thermoanaerobacterium Thermosaccharolyticum and Geobacillus Stearothermophilus in Different Canned Food Matrices

In the canning industry, thermal preservation processes typically are designed based on Clostridium botulinum thermal destruction kinetics. However, some bacteria can still survive, necessitating implementation of stricter timetemperature regimen for sterilization process. The aim of this study was to compare processing effectiveness at F0 (sterilization value) 8 ±1 min from the perspective of the vegetable-based product canning facility, while analyzing the inactivation, viability, and recovery of thermophilic bacteria. Four commercial products [tomato soup and rassolnik soup - acidified food (AF), and mushroom soup and pea porridge - low-acid food (LACF)] with different heat transfer characteristics (convection and conduction) were inoculated with 6.6 log10 spores/ml Geobacillus stearothermophilus LMKK 244 (reported as DSM 6790 and ATCC 10149 in other collections) and 4.810 log spores/ml Thermoanaerobacterium thermosaccharolyticum DSM 571 spore suspensions. Food samples contaminated with bacterial spores were processed in a steam-air retort at 118 °C for 75 min. G. stearothermophilus and T. thermosaccharolyticum growth was not detected in AF samples (pH = 4.4 and 4.5), but was observed in LACF samples (pH = 5.1 and 5.8). Practical evaluation showed that T. thermosaccharolyticum did not survive thermal processing, which was verified using a presence/absence test after incubation at 55 °C. G. stearothermophilus did not survive thermal processing, but recovered in pea porridge (pH = 5.8) during incubation. Our observations showed that food pH is a crucial factor determining microorganism survival during heat treatment and may be used by the vegetable-based product canning facilities to improve the food sterilization conditions.

Purified proteases of two Antarctic bacteria: from screening to characterization

Proteases are widely used in industrial processes, and the discovery of new, more kinetically efficient proteases can have a positive impact on industry. Enzymes from Antarctic microorganisms exhibit cold-adaptive properties, making them useful in biotechnology. The cold and harsh environment of Antarctica makes it a valuable source for new biotechnologically related enzymes. In this study, we characterized two cold-adapted proteases purified from Pseudoalteromonas issachenkonii P14M1-4 and Flavobacterium frigidimaris ANT34-7, isolated from King George Island, Antarctica, and compared these with proteases from the non-cold-adapted bacteria Bacillus licheniformis and Geobacillus stearothermophilus. The best temperature growing conditions were used for protease purification and characterization. The protease from P. issachenkonii P14M1-4 was identified as a 40–43 kDa metal-dependent subtilisin-like serine protease and the protease from F. frigidimaris ANT34-7 was identified as a 28 kDa metalloprotease. The enzymes showed an optimum temperature of between 35°C and 40°C and an optimum pH in the neutral to alkaline range. Their activation energies, catalytic constants and growth capacities at different temperatures categorize them as cold-adapted enzymes. We conclude that the characteristics exhibited by these proteases make them useful for biotechnological purposes requiring high activity at low temperatures. Moreover, to the best of our knowledge, this is the first characterization of a cold-adapted protease from F. frigidimaris.

Public Buses Decontamination by Automated Hydrogen Peroxide Aerosolization System

BACKGROUND: Public transportation has been linked to an increase in the risk of coronavirus disease 2019 transmission. The effective decontamination system using aerosolized hydrogen peroxide can mitigate the transmission risk from using public transportation. AIM: The aim of this study was to develop and validate an effective decontamination system for public transport. METHODS: The experimental research was performed in 13 inter-city public buses. The aerosol generator with ultrasonic atomizer was used in the experiment. The validation process for disinfection was conducted using both a chemical indicator (CI) and spore discs biological indicator (inoculated with 106 Geobacillus stearothermophilus enclosed in glassine envelopes). The CIs and biological indicators were marked by number and placed in nine locations on each bus. The decontamination cycle was developed by analyzed of various aerosolized and decomposition period. Both concentrations of hydrogen peroxide, 5% and 7%, were used for comparison. RESULTS: In an aerosolized period, both concentrations of hydrogen peroxide at 30 min were effective for sporicidal 6-log reductions. The decontamination cycle totaled 100 min, based on a 70 min average decomposition time. CONCLUSIONS: The automated hydrogen peroxide aerosolized system is a highly effective and safe method of decontaminating public buses.

Submerged Bioreactor Production of Geobacillus stearothermophilus ATCC 7953 Spores for Use as Bioindicators to Validate Hydrogen Peroxide Inactivation Processes

In the food and pharmaceutical industries, evaluating the sterilization performance preceding aseptic production processes is of central importance. In the case of hydrogen peroxide sterilization of solid surfaces, bioindicators (BI) consisting of spores of Bacillus atrophaeus or Geobacillus stearothermophilus are used to validate the effectiveness and efficiency of the inactivation procedure. Commercial production of G. stearothermophilus is commonly performed on agar plates, where cultivation and sporulation conditions are not well-defined. Therefore, the produced BI can vary in their resistance, which in turn creates unacceptable uncertainties in the evaluation of aseptic processes. Submerged production in the bioreactor would allow more control over sporulation conditions, while reducing production time, resistance variability, and avoidance of false-positive or false-negative test results. In addition, submerged production of G. stearothermophilus so far was a challenge to achieve sufficiently high spore concentrations for BI production. This study reports on the development of a method for submerged production of G. stearothermophilus spores (pH 7.0, 57 °C, 30% pO2) that can achieve 1.6 × 107 spores/mL with a resistance against 35% H2O2 at 25 °C of D25°C,35% H2O2 = 73 s. This resistance ranks within the range of commercially available BI, making the results directly transferable to industrial applications.