Microencapsulation and sonication: A multiple physical approach to attenuate the probiotic Lacticaseibacillus casei ATCC 393.

Modulation of probiotic performances represents a tool to avoid the probiotic off-flavor in probiotic food. Microencapsulation and sonication were evaluated in slowing down the Lacticaseibacillus casei ATCC 393 induced acidification. Firstly, the influence of alginate concentration and chitosan coating on acidification rate were tested. Microcapsule morphology and the entrapment efficacy were also evaluated. Then, two time of exposure to ultrasound, 6 and 8 min, were applied for L. casei attenuation. Finally, sonicated cells were encapsulated. ΔpH after 6 and 24 h of incubation at 37 °C revealed that chitosan-alginate microcapsules and the 8-min sonicated probiotic presented a significant delayed acidification. When all the systems were compared, the encapsulation of 8-min sonicated L. casei in chitosan-alginate microcapsules significantly improved the results obtained with the single technologies. These results suggest that by modulating the operating parameters and combining these two technologies an increasingly efficient attenuation system can be developed.

Development of Antimicrobial Cellulose Nanofiber-Based Films Activated with Nisin for Food Packaging Applications.

The cellulose nanofiber (CNF) is characterized by the nano-sized (fibers with a diameter between 5 and 20 nm and a length between 2 and 10 µm), flexible and cross-linked structure that confer enhanced mechanical and gas barrier properties to cellulosic fiber-based packaging materials. The purpose of this work was to develop an antimicrobial packaging film by direct mixing nisin with CNF, followed by coating it onto polyethylene (PE), polypropylene (PP), and polylactic acid (PLA) films. The antimicrobial effectiveness of CNF-Nis+PE, CNF-Nis+PP, and CNF-Nis+PLA was investigated both in vitro end in ex vivo tests. In the latter case, challenge test experiments were carried out to investigate the antimicrobial activity of the coupled films of CNF-Nisin+PLA to inhibit the growth of Listeria innocua 1770 during the storage of a meat product. The films were active against the indicator microorganisms Brochothrix thermosphacta and Listeria innocua in in vitro test. Moreover, a reduction in the Listeria population of about 1.3 log cycles was observed immediately after the contact (T0) of the active films with hamburgers. Moreover, when the hamburgers were stored in active films, a further reduction of the Listeria population of about 1.4 log cycles was registered after 2 days of storage. After this time, even though an increase in Listeria load was observed, the trend of the Listeria population in hamburgers packed with active films was maintained significantly lower than the meat samples packed with control films during the whole storage period.

Novel microencapsulated yeast for the primary fermentation of green beer: kinetic behavior, volatiles and sensory profile.

The development of innovative and more cost-effective approaches of making beer throughout continuous fermentation process remains a challenging problem, which is worthy of serious exploration. The current work focuses on the application of a commercial brewing yeast (S. cerevisiae Nottingham Ale), entrapped into chitosan-calcium alginate double layer microcapsules, for the production of a Pale Ale beer. During the primary alcoholic fermentation, the consumption rate of fermentable brewing sugars and dissolved O2, estimated by the Gompertz equation, was halved in the beer obtained by encapsulated yeast in comparison with the free cell. The physical-chemical parameters of beer (i.e. pH, alcohol content, color and bitterness) were not remarkably affected by the different yeast-inoculating form. However, the volatile profiles identified by means of HS-SPME-GC-MS analysis, significantly differed in terms of terpenes, esters and alcohols content, thus proving that the yeast-inoculating form may typify the odor and flavor descriptors of the green beer.

Metabolic Profiling and Cold-Starvation Stress Response of Oxygen-Tolerant Lactobacillus gasseri Strains Cultured in Batch Bioreactor.

Phenotypic and genotypic evidence indicates that many LAB strains can grow in presence of oxygen and can shift from fermentative to aerobic and/or respiratory metabolism. The aerobic and respiratory growth of several LAB species have been studied, allowing the selection of strains showing improved biomass production, long-term survival, and resistance under oxygen and stress conditions. The aim of this work was to observe the adaptation of two Lactobacillus gasseri strains, described in a previous work, to aerobic (air injection) and respiratory (air injection plus hemin and menaquionone) conditions obtained in a batch bioreactor. One strain showed the higher biomass production and oxygen consumption as well as the lower acidification in respiratory condition. Instead, the other one grew better in aerobic condition, even though the higher resistance to cold-starvation stress was registered in respiratory condition. In silico analysis revealed notable differences between AL3 and AL5 genomes and that of the type strain. This work contributes to understanding the adaptation response of lactobacilli to aerobic and respiratory metabolism. We demonstrated that the supposed activation of respiratory metabolism may provide several modifications to cell physiology. These features may be relevant in some technological and health-promoting applications, including starter and probiotic formulations.

Yeast cells in double layer calcium alginate-chitosan microcapsules for sparkling wine production.

This paper focuses on the use of a new type of yeast encapsulation procedure, applying the chitosan-calcium alginate double layer microcapsules, for the production of Riesling sparkling wine. Four different sparkling wines were produced by free or encapsulated yeasts. The four types of yeast used were adapted (Free EtOH-A, Encapsulated EtOH-A) and non-adapted to ethanol (Free, Encapsulated). The different yeast-inoculating formats had a significant impact on oxygen consumption and pressure increase rate in the bottle during the prise de mousse. Similarly to the free form, encapsulated yeast successfully completed the secondary fermentation. After an ageing period of 6 months, volatiles and sensory profiles of sparkling wines were compared. Although, some differences in volatile profiles were found among samples, sparkling wines produced by Encapsulated EtOH-A showed sensory properties, in terms of aroma, taste and body, similar to those produced by free yeast (both adapted and non-adapted to ethanol).

Adaptation to Aerobic Environment of Lactobacillus johnsonii/gasseri Strains.

Oxygen is considered one of the main factors affecting probiotic bacteria survival due to the induction of oxidative damages caused by the action of reactive oxygen species (ROS). It has been shown that oxidative stress resistance in lactic acid bacteria is strongly dependent on the type of cell metabolism. Shift from fermentative to respiratory metabolism (through the addition of heme and menaquinone and in presence of oxygen) was associated to increase in biomass, long-term survival, and production of antioxidant enzymes. The aim of this work was to investigate the effect of aerobic (presence of oxygen) and respiratory (presence of oxygen, heme, and menaquinone) cultivation on the growth kinetic, catalase production, oxygen uptake, and oxidative stress response of Lactobacillus johnsonii/gasseri strains previously isolated from infant feces. Seven strains showed to consume oxygen under aerobic and respiratory conditions. The strain AL5 showed a catalase activity in both growth conditions, while AL3 showed this activity only in respiratory condition. Respiratory condition improved their tolerance to oxidative compounds (hydrogen peroxide and ROS generators) and further they showed promising probiotic features. The exploration of respiratory competent phenotypes with probiotic features may be extremely useful for the development of competitive starter or probiotic cultures.

Draft Genome Sequences of the Aerobic Strains Lactobacillus gasseri AL3 and AL5.

Adaptation to the aerobic environment has been investigated in heterofermentative lactobacilli, while data on how homofermentative lactobacilli adapt to oxygen are limited. We report here the draft genome sequences of the aerobic strains Lactobacillus gasseri AL3 and AL5 that allow an in-depth investigation of the genes involved in oxidative metabolism and the stress response.