In vivo Assessment of Cold Atmospheric Pressure Plasma Technology on the Bioactivity of Spirulina.

The present study challenges the in vivo assessment of cold atmospheric pressure plasma (CAPP) technology on the bioactive activity (antioxidant/antiaging and antimicrobial potential) of Spirulina powder, using Caenorhabditis elegans as an animal model. Surface microdischarge cold atmospheric pressure plasma (SMD-CAPP) treatment was 3.3 W discharge power for 7 min. C. elegans lifespan and egg laying were used as indicators of antioxidant/antiaging potential of Spirulina (1 mg/mL), when grown with Spirulina CP-treated [E_SCP] and untreated [E_S], compared with a control [E_0] (non-supplemented with Spirulina). According to our results, under both Spirulina supplemented media [E_SCP and E_S] and for the first 17 days, nematodes experienced an increase in lifespan but without significant differences (p > 0.05) between control and Spirulina CP-treated. Regarding the in vivo assay of the antimicrobial potential of Spirulina against Salmonella enterica serovar Typhimurium (infected worms), no significant differences (p > 0.05) were found between the three exposure scenarios (control [S_0]; Spirulina supplemented media [S_S]; CP-treated Spirulina supplemented media [S_SCP]). According to present results, CAPP-treatment do not influence negatively the lifespan of C. elegans but a reduction in the Spirulina antiaging potential was found. No in vivo modifications in antimicrobial activity seem to be linked to CAPP-processed Spirulina.

Caenorhabditis elegans as an in vivo model to assess fucoidan bioactivity preventing Helicobacter pylori infection.

Currently, Helicobacter pylori is the unique biological carcinogenic agent. The search for antimicrobial alternatives to antibiotics against this pathogen has been categorized as a priority due to the drastic failure associated with current applied antibiotic therapy. The present study assessed the bioactive antimicrobial capability of fucoidan ("Generally Recognized as Safe" approval - European Commission December 2017) from different species of Phaeophyceae algae (Fucus vesiculosus, Undaria pinnatifida, Macrocystis pyrifera) against H. pylori. All the studied fucoidans showed bacteriostatic and bactericidal effects at the studied concentrations [5-100] µg ml-1 and exposure times [0-7 days]. The most effective anti-H. pylori fucoidan was validated in Caenorhabditis elegans as an in vivo model. C. elegans feed was supplemented with Undaria pinnatifida [0-100] µg ml-1 fucoidan, resulting in a significant improvement in lifespan, lowered H. pylori concentration in the digestive tract, and increased egg-laying pattern. New research lines proposing this compound as an active agent in nutraceutical and preventive novel therapies should be opened.

Helicobacter Pylori Detection in Shellfish: A Real-Time Quantitative Polymerase Chain Reaction Approach.

Shellfish is a highly valuated natural food product that is usually consumed minimally processed. Some foodborne pathogens have been associated to marine products and isolated from aquatic environments. Helicobacter pylori emerges as one of the most concerning human pathogens associated to water and, thereby, it could be present in raw and slightly treated marine food products. The present research work aimed to detect the presence of H. pylori in Spanish commercial samples of shellfish (mussels, clams, and cockles) by means of a quantitative real-time polymerase chain reaction (qPCR) approach based on the vacuolating cytotoxin A (vacA) gene specificity. Putative H. pylori amplicons were confirmed by sequencing. qPCR was positive for 12 out of the 100 samples, being 67% (8/12) from mussels, 25% (3/12) from clams, and only 8% (1/12) from cockles. After sequencing, three of the amplicons showed 97-99% homology with the H. pylori vacA gene. Quantitative results indicate that the levels of contamination remained below 102 log10 colony forming units per mL (CFU/mL). The present research shows for the first time the effectiveness of the optimized qPCR in the identification of potentially H. pylori contaminated shellfish products. Our results confirm the presence of H. pylori in shellfish from the Spanish western seacoast and verify the possible relationship between the presence of H. pylori in seawater and the role of contaminated seafoods as vehicles of H. pylori entrance into the food chain.

Use of the modified Gompertz equation to assess the Stevia rebaudiana Bertoni antilisterial kinetics.

In order to assess the antibacterial activity of Stevia rebaudiana Bertoni (Stevia), Listeria innocua growth was characterized at 37 °C, in reference medium supplemented with a leaf infusion, a crude extract, and a steviol glycosides purified extract. Experimental data were fitted to the modified Gompertz model and the antibacterial activity of Stevia was determined based on the lag time (λ) and the maximum growth rate (µmax) reached, depending on the incubation conditions. As the leaf infusion showed the most marked elongation of λ and the most marked µmax reduction, its antimicrobial effect was evaluated at different concentrations, at 37, 22 and 10 °C. According to the results obtained, in general, the lower the temperature or the higher the Stevia concentration, the longer the λ and the lower the µmax, statistically significant being the effect of reducing temperature from 37 or 22 to 10 °C, the effect of increasing Stevia concentration from 0 or 0.5 to 1.5 or 2.5% (w/v), at 37 °C, and the elongation of λ observed in presence of 1.5 and 2.5% (w/v) of Stevia, at 22 °C. These results show that Stevia could be a bacterial growth control measure if a cold chain failure occurs.

Inactivation kinetics of spores of Bacillus cereus strains treated by a peracetic acid-based disinfectant at different concentrations and temperatures.

The purpose of this study was to assess the effect of a commercial peracetic acid-based disinfectant against spores of Bacillus cereus, to identify the most influential factor for the final number of microorganisms after different disinfection procedures, and to evaluate the nature of the inactivation kinetics. The spores of four different strains of B. cereus (DSM 318, 4312, 4313, and 4384) were treated with five different disinfectant concentrations (0.25%, 0.5%, 1.0%, 1.5%, and 2.0% [w/v]) at three different temperatures (10°C, 15°C, and 20°C) with or without protein load. A higher temperature and PES 15/23 concentration resulted in a higher inactivation. Inactivation of B. cereus strain 4312 was around 2 log10 cycles at 10°C and around 7 log10 at 20°C (conc=1% [w/v] PAA; t=60 min; without protein). The protein load at higher concentrations did not significantly reduce the efficacy of the disinfectant (p>0.05). This article indicates the applicability of the Weibull model to fit the B. cereus disinfectant survival curves. A Monte Carlo simulation was used to carry out a sensitivity analysis, which revealed the most influential factors affecting the final number of microorganisms after the disinfection process.

Monte Carlo simulation as a method to determine the critical factors affecting two strains of Escherichia coli inactivation kinetics by high hydrostatic pressure.

The survival curves of two strains of Escherichia coli (ATCC 4157 and ATCC 23716) were obtained by high hydrostatic pressure (HHP) at four pressure levels (100, 200, 300, and 400 MPa) in an orange juice-skim milk (OJ-SM) mixed beverage. The effect of suspension medium pH on HHP inactivation was studied at three pH levels: 4, 4.5, and 5. The Weibull model provided an accurate fit to the data according to adjusted-regression coefficients (adjusted-R(2)) [0.860-0.996], root mean square error values [0.001-0.150], and accuracy factor [1.001-1.125] values. For sensitivity analysis, Monte Carlo simulation was run to determine the most influential factors affecting HHP inactivation of E. coli strains. According to these results, the most influential factor for E. coli (ATCC 4157) HHP inactivation was the pH of the OJ-SM beverage at low HHP intensity, whereas pressure was more influential under strong HHP treatment conditions. Concerning E. coli K-12 (ATCC 23716), pressure (P) was the most influential parameter independently of the pH of OJ-SM beverage. The higher the P, the higher the inactivation achieved. The sensitivity analysis indicated greater E. coli K-12 (ATCC 23716) resistance to the HHP processing, possibly because of greater acidity tolerance.

Synergistic effect of high hydrostatic pressure and natural antimicrobials on inactivation kinetics of Bacillus cereus in a liquid whole egg and skim milk mixed beverage.

An in-depth study was conducted in order to extend the storage life of a liquid whole egg-skim milk (LWE-SM) mixed beverage to enhance its safety and the safety of related beverages. Bacillus cereus vegetative cells (1 x 10(8) colony-forming units [CFU]/mL) were inoculated in LWE-SM beverages with or without natural antimicrobial supplements: flavonol rich-cocoa powder (cocoanOX 12%, CCX) (700 ppm), vanillin (700 ppm), anise (700 ppm), and cinnamon (700 ppm). B. cereus cells were maintained at 10 degrees C for 10 days in the different beverages to test the bacteriostatic or inhibitory effect of the aforementioned ingredients. Beverages were treated with high hydrostatic pressure (HHP) technology and stored at 10 degrees C for 15 days after treatment. All natural antimicrobials reduced the micro(max) values and increased the lag phase time of B. cereus, and Gompertz growth curves showed different inhibitory effects depending on the substance. The maximum inhibitory effect (1.330 log cycle reduction) was achieved in LWE-SM-cinnamon-supplemented beverage. The maximum inactivation achieved by HHP in LWE-SM beverage was a reduction of around 3.89 +/- 0.25 log cycles at 300 MPa for 12 minutes. When supplemented beverages were treated under the same conditions, enhanced inactivation levels were achieved. This increased inactivation can be attributed to a synergistic effect when the LWE-SM was supplemented with flavonol-rich cocoa powder, cinnamon, and vanillin. The maximum synergistic effect was observed in LWE-SM-CCX-supplemented beverage. During the refrigerated storage of B. cereus HHP-treated cells in beverages to which antimicrobials had been added, the inhibitory effect was dependent on the previously applied pressure level.