Impact of pulsed-electric field and high-voltage electrical discharges on red wine microbial stabilization and quality characteristics.

AIMS: In this study, pulsed-electric fields (PEF) and high-voltage electrical discharges (HVED) are proposed as new techniques for the microbial stabilization of red wines before bottling. The efficiency of the treatment was then evaluated. METHODS AND RESULTS: PEF and HVED-treatments have been applied to wine for the inactivation of Oenococcus oeni CRBO 9304, O. oeni CRBO 0608, Pediococcus parvulus CRBO 2.6 and Brettanomyces bruxellensis CB28. Different treatment times (1, 2, 4, 6, 8 and 10 ms) were used at 20 kV cm(-1) for the PEF treatments and at 40 kV for the HVED treatments, which correspond to applied energies from 80 to 800 kJ l(-1) . The effects of the treatments on the microbial inactivation rate and on various characteristics of red wines (phenolic composition, chromatic characteristics and physico-chemical parameters) were measured. CONCLUSIONS: The application of PEF or HVED treatments on red wine allowed the inactivation of alteration yeasts (B. bruxellensis CB28) and bacteria (O. oeni CRBO 9304, O. oeni CRBO 0608 and P. parvulus CRBO 2.6). The electric discharges at 40 kV were less effective than the PEF even after 10 ms of treatments. Indeed, 4 ms of PEF treatment at 20 kV cm(-1) were sufficient to inactivate all micro-organisms present in the wines. Also, the use of PEF had no negative impact on the composition of wines compared to the HVED treatments. Contrary to PEF, the phenolics compounds were degraded after the HVED treatment and the physico-chemical composition of wine were modified with HVED. SIGNIFICANCE AND IMPACT OF THE STUDY: PEF technology seems to be an interesting alternative to stabilize microbiologically wines before bottling and without modifying their composition. This process offers many advantages for winemakers: no chemical inputs, low energy consumption (320 kJ l(-1) ), fast (treatment time of 4 ms) and athermal (ΔT ≈ 10°C).

Pediococcus pentosaceus SB83 as a potential probiotic incorporated in a liquid system for vaginal delivery.

Pediococcus pentosaceus SB83 is a bacteriocinogenic culture having potential use as a vaginal probiotic. The objective of this study was to evaluate the behaviour of P. pentosaceus SB83 incorporated in a liquid system for eventually formulating a gel for vaginal delivery. The vaginal probiotic incorporated into glycerol was able to survive in simulated vaginal fluid at normal vaginal pH (4.2) and at pH of vaginal infections (5.5 and 6.5). The probiotic can be stored at 4 °C for a longer period of time than at room temperature, however, after 13 weeks of storage at low temperature, there was a total loss of viable cells. The probiotic strain incorporated into glycerol showed bacteriocinogenic activity in simulated vaginal fluid, although the antimicrobial activity against Listeria monocytogenes declined during storage. This study showed the behaviour of P. pentosaceus SB83 in glycerol and in simulated vaginal fluid. However, it is necessary to optimise the formulation to produce an actively probiotic vaginal gel.

Characterisation of the heat-stable bacteriocin-producing and vancomycin-sensitive Pediococcus pentosaceus CFR B19 isolated from beans.

A bacteriocin-producing lactic culture with antilisterial activity was isolated from beans and identified as Pediococcus pentosaceus CFR B19. It was able to grow and produce bacteriocin at 41 °C but not at 50 °C. This isolate was found to be sensitive to vancomycin and produced heat-stable (at 121 °C for 15 min) bacteriocin. Molecular weight of the purified bacteriocin was found to be ∼4.8 kDa. This isolate can be used as a starter culture or co-culture in fermented milk products and the bacteriocin can be used as a natural preservative in various food products.

Exploring the use of natural antimicrobial agents and pulsed electric fields to control spoilage bacteria during a beer production process.

Different natural antimicrobials affected viability of bacterial contaminants isolated at critical steps during a beer production process. In the presence of 1 mg/ml chitosan and 0.3 mg/ml hops, the viability of Escherichia coli in an all malt barley extract wort could be reduced to 0.7 and 0.1% respectively after 2 hour- incubation at 4 degrees C. The addition of 0.0002 mg/ml nisin, 0.1 mg/ml chitosan or 0.3 mg/ml hops, selectively inhibited growth of Pediococcus sp. in more than 10,000 times with respect to brewing yeast in a mixed culture. In the presence of 0.1 mg ml chitosan in beer, no viable cells of the thermoresistant strain Bacillus megaterium were detected. Nisin, chitosan and hops increased microbiological stability during storage of a local commercial beer inoculated with Lactobacillus plantarum or Pediococcus sp. isolated from wort. Pulsed Electric Field (PEF) (8 kV/cm, 3 pulses) application enhanced antibacterial activity of nisin and hops but not that of chitosan. The results herein obtained suggest that the use of these antimicrobial compounds in isolation or in combination with PEF would be effective to control bacterial contamination during beer production and storage.

Exploring the use of natural antimicrobial agents and pulsed electric fields to control spoilage bacteria during a beer production process / Exploración del uso de agentes antimicrobianos naturales y de campos eléctricos pulsantes para el control de bacterias contaminantes durante el proceso de elaboración de cerveza

Different natural antimicrobials affected viability of bacterial contaminants isolated at critical steps during a beer production process. In the presence of 1 mg/ml chitosan and 0.3 mg/ml hops, the viability of Escherichia coli in an all malt barley extract wort could be reduced to 0.7 and 0.1% respectively after 2 hour- incubation at 4 °C. The addition of 0.0002 mg/ml nisin, 0.1 mg/ml chitosan or 0.3 mg/ml hops, selectively inhibited growth of Pediococcus sp. in more than 10,000 times with respect to brewing yeast in a mixed culture. In the presence of 0.1mg ml chitosan in beer, no viable cells of the thermoresistant strain Bacillus megaterium were detected. Nisin, chitosan and hops increased microbiological stability during storage of a local commercial beer inoculated with Lactobacillus plantarum or Pediococcus sp. isolated from wort. Pulsed Electric Field (PEF) (8 kV/cm, 3 pulses) application enhanced antibacterial activity of nisin and hops but not that of chitosan. The results herein obtained suggest that the use of these antimicrobial compounds in isolation or in combination with PEF would be effective to control bacterial contamination during beer production and storage.

Diferentes antimicrobianos naturales disminuyeron la viabilidad de bacterias contaminantes aisladas en etapas críticas del proceso de producción de cerveza. En un extracto de malta, el agregado de 1 mg/ml de quitosano y de 0,3 mg ml de lúpulo permitió reducir la viabilidad de Escherichia coli a 0,7 y 0,1%, respectivamente, al cabo de 2 horas de incubación a 4 °C. El agregado de 0,0002 mg/ml de nisina, 0,1 mg/ml de quitosano o de 0,3 mg/ml de lúpulo inhibió selectivamente (10.000 veces más) el crecimiento de Pediococcus sp. respecto de la levadura de cerveza en un cultivo mixto. El agregado de 0,1 mg/ml de quitosano permitió disminuir la viabilidad de una cepa bacteriana termorresistente, Bacillus megaterium, hasta niveles no detectables. Por otra parte, el agregado de nisina, quitosano y lúpulo aumentó la estabilidad microbiológica durante el almacenamiento de cervezas inoculadas con Lactobacillus plantarum y Pediococcus sp. aislados de mosto de cerveza. La aplicación de campos eléctricos pulsantes (CEP) (3 pulsos de 8kV/cm) aumentó el efecto antimicrobiano de la nisina y del lúpulo, pero no el del quitosano. Los resultados obtenidos indicarían que el uso de antimicrobianos naturales en forma individual o en combinación con CEP puede constituir un procedimiento efectivo para el control de la contaminación bacteriana durante el proceso de elaboración y almacenamiento de la cerveza.

Partial characterization of bacteriocins produced by human Lactococcus lactis and Pediococccus acidilactici isolates.

AIMS: The aim of this study was to isolate bacteriocin-producing lactic acid bacteria (LAB) from human intestine. METHODS AND RESULTS: A total of 111 LAB were isolated from human adult stool and screened for their bacteriocin production. Neutralized cell-free supernatants from Lactococcus lactis subsp. lactis MM19 and Pediococcus acidilactici MM33 showed antimicrobial activity. The antimicrobials in the supernatant from a culture of L. lactis inhibited Enterococcus faecium, various species of Lactobacillus and Staphylococcus aureus; while those in the supernatant from a culture of P. acidilactici inhibited Enterococcus spp., some lactobacilli and various serotypes of Listeria monocytogenes. The antimicrobial metabolites were heat-stable and were active over a pH range of 2-10. The antimicrobial activities of the supernatants of both bacteria were inhibited by many proteases but not by catalase. The plate overlay assay allowed an approximation of size between 3.5 and 6 kDa for both antimicrobial substances. CONCLUSIONS: As the antagonistic factor(s) produced by L. lactis MM19 and P. acidilactici MM33 were sensitive to proteolytic enzymes, it could be hypothesized that bacteriocins were involved in the inhibitory activities. Inhibition spectrum and biochemical analysis showed that these bacteria produced two distinct bacteriocins. SIGNIFICANCE AND IMPACT OF THE STUDY: We are the first to isolate bacteriocin-producing strains of Pediococcus and Lactococcus from human intestine. These strains might be useful for control of enteric pathogens.

Inactivation of microorganisms with microwaves at reduced temperatures.

We developed a pilot-plant nonthermal flow process using microwave energy to inactivate microorganisms. The process consists of multiple passes through the microwave generator. Each passed material goes to a receiving tank for subsequent passes. The flow rate was 0.96 to 1.26 kg/min and the dwell time per pass was 1.1 to 1.5 min. Five passes were used. The microwave energy is instantaneously and simultaneously applied to the system, and thermal energy is removed by a cooling tube within the process line in the microwave generator. The cooling tube maintains the temperature below 40 degrees C. There was significant reduction in microorganisms in water, 10% glucose solution, and apple juice, and in yeast in beer. There was a slight decrease in microorganisms in tomato juice, pineapple juice, apple cider, and beer; and no effect in skim milk.