Inactivation of Bacillus cereus by Na-chlorophyllin-based photosensitization on the surface of packaging.

AIMS: This study was focused on the possibility to inactivate food-borne pathogen Bacillus cereus by Na-chlorophyllin (Na-Chl)-based photosensitization in vitro and after attachment to the surface of packaging material. METHODS AND RESULTS: Bacillus cereus in vitro or attached to the packaging was incubated with Na-Chl (7·5×10(-8) to 7·5×10(-5) mol l(-1) ) for 2-60min in phosphate buffer saline. Photosensitization was performed by illuminating cells under a light with a λ of 400nm and an energy density of 20mW cm(-2) . The illumination time varied 0-5min and subsequently the total energy dose was 0-6J cm(-2) . The results show that B. cereus vegetative cells in vitro or attached to the surface of packaging after incubation with 7·5×10(-7) mol l(-1) Na-Chl and following illumination were inactivated by 7log. The photoinactivation of B. cereus spores in vitro by 4log required higher (7·5×10(-6) mol l(-1) ) Na-Chl concentration. Decontamination of packaging material from attached spores by photosensitization reached 5log at 7·5×10(-5) mol l(-1) Na-Chl concentration. Comparative analysis of different packaging decontamination treatments indicates that washing with water can diminish pathogen population on the surface by <1log, 100ppm Na-hypochlorite reduces the pathogens about 1·7log and 200ppm Na-hypochlorite by 2·2log. Meanwhile, Na-Chl-based photosensitization reduces bacteria on the surface by 4·2 orders of magnitude. CONCLUSIONS: Food-borne pathogen B. cereus could be effectively inactivated (7log) by Na-Chl-based photosensitization in vitro and on the surface of packaging material. Spores are more resistant than vegetative cells to photosensitization-based inactivation. Comparison of different surface decontamination treatments indicates that Na-Chl-based photosensitization is much more effective antibacterial tool than washing with water or 200ppm Na-hypochlorite. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data support the idea that Na-Chl-based photosensitization has great potential for future application as an environment-friendly, nonthermal surface decontamination technique.

Inactivation of food pathogen Bacillus cereus by photosensitization in vitro and on the surface of packaging material.

AIMS: The study was focused on the possibility to inactivate food pathogen Bacillus cereus by 5-aminolevulinic acid (ALA) - based photosensitization in vitro and after adhesion on the surface of packaging material. METHODS AND RESULTS: Bacillus cereus was incubated with ALA (3-7.5 mmol l(-1)) for 5-60 min in different environment (PBS, packaging material and wheat grains) and afterwards illuminated with visible light. The light source used for illumination emitted light at lambda = 400 nm with energy density at the position of the cells, 20 mW cm(-2). The illumination time varied from 0 to 20 min, and subsequently a total energy dose was between 0 and 24 J cm(-2). The obtained results indicate that B. cereus after the incubation with 3-7.5 mmol l(-1) ALA produces suitable amounts of endogenous photosensitizers. Following illumination, micro-organism inactivated even by 6.3 log. The inactivation of B. cereus after adhesion on the surface of food packaging by photosensitization reached 4 log. It is important to note that spores of B. cereus were susceptible to this treatment as well; 3.7-log inactivation in vitro and 2.7-log inactivation on the surface of packaging material were achieved at certain experimental conditions. CONCLUSIONS: Vegetative cells and spores of Gram-positive food pathogen B. cereus were effectively inactivated by ALA-based photosensitization in vitro. Moreover, the significant inactivation of B. cereus adhered on the surface of packaging material was observed. It was shown that photosensitization-based inactivation of B. cereus depended on the total light dose (illumination time) as well as on the amount of endogenous porphyrins (initial ALA concentration, time of incubation with ALA). SIGNIFICANCE AND IMPACT OF THE STUDY: Our previous data, as well as the one obtained in this study, support the idea that photosensitization with its high selectivity, antimicrobial efficiency and nonthermal nature could serve in the future for the development of completely safe, nonthermal surface decontamination and food preservation techniques.

Modelling the photosensitization-based inactivation of Bacillus cereus.

AIMS: To study and to develop a model for the photo-destruction of the foodborne pathogen Bacillus cereus, initially treated with a precursor of endogenous photosensitizers (5-aminolevulinic acid, ALA). MATERIALS AND METHODS: The cells were incubated in the presence of ALA (3 or 7.5 mmol l(-1)) for incubation times ranging from 2 to 60 min, inoculated onto the surface of LB Agar plates and submitted to light irradiation. The Weibull model was used to describe the survival curves of B. cereus. Quadratic equations were used to describe the effects of ALA concentration and incubation time on the Weibull model parameters. RESULTS: ALA-based photosensitization proved to be an effective tool for inactivation of B. cereus. The decrease in viable counts observed after 20 min of irradiation, ranged from 4 to 6 log CFU g(-1). CONCLUSIONS: The developed model proved to be a parsimonious and robust solution to describe the observed data. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrates the effectiveness of photosensitization on B. cereus on agar plates. The model developed may be useful to optimize inactivation treatments by photosensitization.

Novel approach to control Salmonella enterica by modern biophotonic technology: photosensitization.

AIMS: Salmonellosis is one of the most common foodborne diseases in the world. The aim of this study was to evaluate the antibacterial efficiency of 5-aminolevulinic acid (ALA) based photosensitization against one of food pathogens Salmonella enterica. METHODS AND RESULTS: Salmonella enterica was incubated with ALA (7.5 mmol l(-1)) for 1-4 h and afterwards illuminated with visible light. The light source used for illumination of S. enterica emitted light lambda = 400 nm with energy density 20 mW cm(-2). The illumination time varied from 0 to 20 min and subsequently a total energy dose reached 0-24 J cm(-2). The data obtained indicate that S. enterica is able to produce endogenous photosensitizer PpIX when incubated with ALA. Remarkable inactivation of micro-organisms can be achieved (6 log) after photosensitization. It is obvious that photosensitization-based inactivation of S. enterica depends on illumination as well as incubation with ALA time. CONCLUSION: ALA-based photosensitization can be an effective tool against multi-drug resistant Gram-negative bacteria S. enterica serovar Typhimurium. SIGNIFICANCE AND IMPACT OF THE STUDY: Experimental data and mathematical evaluations support the idea that ALA-based photosensitization can be a useful tool for the development of nonthermal food preservation technology in future.

Advanced high-power pulsed light device to decontaminate food from pathogens: effects on Salmonella typhimurium viability in vitro.

AIMS: The aim of this study was to construct an advanced high-power pulsed light device for decontamination of food matrix and to evaluate its antibacterial efficiency. Key parameters of constructed device-emitted light spectrum, pulse duration, pulse power density, frequency of pulses, dependence of emitted spectrum on input voltage, irradiation homogenicity, possible thermal effects as well as antimicrobial efficiency were evaluated. METHODS AND RESULTS: Antimicrobial efficiency of high-power pulsed light technique was demonstrated and evaluated by two independent methods - spread plate and Miles-Misra method. Viability of Salmonella typhimurium as function of a given light dose (number of pulses) and pulse frequency was examined. According to the data obtained, viability of Salmonella typhimurium reduced by 7 log order after 100 light pulses with power density 133 W cm(-2). In addition, data indicate, that the pulse frequency did not influence the outcome of pathogen inactivation in the region 1-5 Hz. Moreover, no hyperthermic effect was detected during irradiation even after 500 pulses on all shelves with different distance from light source and subsequently different pulse power density (0-252 W cm(-2)). CONCLUSION: Newly constructed high-power pulsed light technique is effective nonthermal tool for inactivation of Salmonella typhimurium even by 7 log order in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: Novel advanced high-power pulsed light device can be a useful tool for development of nonthermal food decontamination technologies.