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Decontamination of Bacillus subtilis Spores in a Sealed Package Using a Non-thermal Plasma System

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Book cover Plasma for Bio-Decontamination, Medicine and Food Security

Abstract

The safety of packaged food and medical devices is a major concern to consumers and government officials. Recent inventions (PK-1 and PK-2) based on the principles of non-thermal, atmospheric plasma has shown significant reduction in bacterial contamination inside a sealed package. The objective of this study was to evaluate the PK-1 and PK-2 systems in the reduction of Bacillus subtilis spores using packages containing air or modified atmosphere (MA) gas (65% O2/30% CO2/5% N2). The experimental design consisted of the following parameters: (1) two voltage conditions: 13.5 kV with 1.0 cm electrode gap (PK-1) and 80 kV with 4.5 cm electrode gap (PK-2), (2) two treatment conditions: inside and outside the field of ionization, (3) PK-1 and PK-2 optimized treatment times: 300 and 120 s, respectively, and (4) two package gas types: air and modified atmosphere (MA) gas (65% O2/30% CO2/5% N2). Measurements included: (1) bacterial reductions of Bacillus subtilis var. niger (B. atrophaeus), (2) ozone, nitrous oxides (NOx), and carbon monoxide concentrations, and (3) relative humidity. Bacillus subtilis (1.7 × 106/strip) were loaded into sterile uncovered petri dishes and treated with ionization generated in packages using air or MA gas blend. Samples were treated for 300 s (PK-1) or 120 s (PK-2) and stored at room ­temperature for 24 h. Results documented relative humidity (RH) ranged from 20% to 30%. After 300 s of PK-1 treatment (13.5 kV/44 W/1.0 cm gap), ozone concentrations were 6,000 ppm (air) and 7,500 ppm (MA). After 120 s of PK-2 treatment (80 kV/150 W/4.5 cm), ozone concentrations were 7,500 ppm (air) and 12,000 ppm (MA). Ozone and NOx concentrations were non-detect (ND) after 24 h. PK-1 carbon monoxide levels were <20 ppm (air) and <100 ppm (MA) after 24 h. The PK-2 carbon monoxide levels were <20 ppm (air) and <400 ppm (MA) after 24 h. Treatments showed reductions in spores of greater than 6 log10 after 24 h. Reductions were maintained without additional re-growth at 72 h. These results indicate that the PK-1 and PK-2 systems have the capacity to reduce Bacillus subtilis spores in an in-package ionization process.

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Correspondence to Kevin M. Keener .

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Keener, K.M. et al. (2012). Decontamination of Bacillus subtilis Spores in a Sealed Package Using a Non-thermal Plasma System. In: Machala, Z., Hensel, K., Akishev, Y. (eds) Plasma for Bio-Decontamination, Medicine and Food Security. NATO Science for Peace and Security Series A: Chemistry and Biology. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-2852-3_34

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