Probiotic Potential and Application of Indigenous Non-Starter Lactic Acid Bacteria in Ripened Short-Aged Cheese.

There are massive sources of lactic acid bacteria (LAB) in traditional dairy products. Some of these indigenous strains could be novel probiotics with applications in human health and supply the growing needs of the probiotic industry. In this work, were analyzed the probiotic and technological properties of three Lactobacilli strains isolated from traditional Brazilian cheeses. In vitro tests showed that the three strains are safe and have probiotic features. They presented antimicrobial activity against pathogenic bacteria, auto-aggregation values around 60%, high biofilm formation properties, and a survivor of more than 65% to simulated acid conditions and more than 100% to bile salts. The three strains were used as adjunct cultures separately in a pilot-scale production of Prato cheese. After 45 days of ripening, the lactobacilli counts in the cheeses were close to 8 Log CFU/g, and was observed a reduction in the lactococci counts (around -3 Log CFU/g) in a strain-dependent manner. Cheese primary and secondary proteolysis were unaffected by the probiotic candidates during the ripening, and the strains showed no lipolytic effect, as no changes in the fatty acid profile of cheeses were observed. Thus, our findings suggest that the three strains evaluated have probiotic properties and have potential as adjunct non-starter lactic acid bacteria (NSLAB) to improve the quality and functionality of short-aged cheeses.

Brazilian indigenous nonstarter lactic acid bacteria enhance the diversification of volatile compounds in short-aged cheese.

There is growing interest in using autochthonous lactic acid bacteria (LAB) that provide unique sensory characteristics to dairy products without affecting their safety and quality. This work studied the capacity of three Brazilian indigenous nonstarter LABs (NSLAB) to produce biogenic amines (BAs) and evaluated their effect on the volatile organic compounds (VOCs), microbial LAB communities, and physicochemical profile of short-aged cheese. Initially, the strain's potential for biosynthesis of BAs was assessed by PCR and in vitro assays. Then, a pilot-scale cheese was produced, including the NSLAB, and the microbial and VOC profiles were analyzed after 25 and 45 days of ripening. As a results, the strains did not present genes related to relevant BAs and did not produce them in vitro. During cheese ripening, the Lactococci counts were reduced, probably in the production of alcohols and acid compounds by the NSLAB. Each strain produces a unique VOC profile that changes over the ripening time without the main VOCs related to rancid or old cheese. Particularly, the use of the strain Lacticaseibacillus. paracasei ItalPN16 resulted in production of ester compounds with fruity notes. Thus, indigenous NSLAB could be a valuable tool for the enhancement and diversification of flavor in short-aged cheese.

Draft Genome Sequence and Comparative Genome Analysis Reveal Potential Functional Properties in Lacticaseibacillus paracasei ItalPN16.

Nowadays, there is a great interest on rapid and effective methods for initial identification of probiotic bacteria. In this work, potential probiotic features of the lactic acid bacteria strain ItalPN16 isolated from a traditional Brazilian cheese were studied using bioinformatic tools. The complete genome sequence was obtained, and in silico analyses were carried out to identify the strain and its potential probiotic properties. The sequenced genome (3.02 Mb) presented 3126 protein-coding sequences distributed on 244 SEED subsystems, classifying the strain as nomadic lactobacilli. Phylogenetic and ANI analyses allowed to locate the ItalPN16 strain as a member of the Lacticaseibacillus paracasei group, due to the highest number of orthologous genes in common with reference L. paracasei strains (>98%). In silico analyses revealed the presence of CDSs related to microbe-host interactions, such as adhesion proteins and exopolysaccharide biosynthesis genes. The comparative analysis reveals the presence of a strain-specific glycosyl transferases, compared with other three L. paracasei strains and a high level of protein expression (92%) with the probiotic L. paracasei BL29. The results obtained here indicated interesting probiotic features of the strain L. paracasei ItalPN16 that could favor a future application in the food industry.

Improved functional properties of the potential probiotic Lacticaseibacillus paracasei ItalPN16 growing in cheese whey.

The production of probiotic bacteria requires specific and expensive culture media for maintain their viability and metabolic response during gastro-intestinal transit and cell adhesion process. The aim of this study was to compare the ability of the potential probiotic Laticaseibacillus paracasei ItalPN16 to grow in plain sweet whey (SW) and acid whey (AW), evaluating changes in some probiotic properties related to the culture media. Pasteurized SW and AW were suitable media for L. paracasei growth, since counts above 9 Log CFU/ml were achieved using <50% of the total sugars in both whey samples after 48 h at 37°C. The L. paracasei cells obtained from AW or SW cultures showed increased resistance to pH 2.5 and 3.5, higher autoaggregation, and lower cell hydrophobicity, as compared with the control of MRS. SW also improved the biofilm formation ability and cell adhesion capability to Caco-2 cells. Our results indicate that the L. paracasei adaptation to the SW conditions, inducing metabolic changes that improved its stability to acid stress, biofilm formation, autoaggregation, and cell adhesion properties, which are important functional probiotic properties. Overall, the SW could be considered as low-cost culture medium for sustainable biomass production of L. paracasei ItalPN16.

Carvacrol encapsulation into nanoparticles produced from chia and flaxseed mucilage: Characterization, stability and antimicrobial activity against Salmonella and Listeria monocytogenes.

Carvacrol is a natural antimicrobial with excellent antimicrobial properties against several foodborne pathogens. Encapsulation can increase carvacrol stability and solubility, and mask its pronounced odor. Mucilages have been studied as wall material for nanoparticles due to their high retention capacity of bioactive compounds and ease of chemical modifications to improve their stability. In this study, 1.67 mg/mL of carvacrol encapsulated into chia mucilage nanoparticles (CMNP) and flaxseed mucilage nanoparticles (FMNP) were produced by high-energy emulsification technique and tested against Listeria monocytogenes and Salmonella. Encapsulation efficiency around 98% of carvacrol was obtained for both formulations. CMNP showed a diameter size of 179 nm and zeta potential of -11.4 mV. Bacterial Inactivation Concentration (BIC) of CMNP was 0.42 mg/mL against Salmonella and 0.83 mg/mL against L. monocytogenes. FMNP showed diameter size of 165.3 nm and zeta potential of -12.6 mV. BIC of FMNP was 0.83 mg/mL against both microorganisms. Scanning electron microscopy analysis showed that the nanoparticles are spherically shaped. Concentrations of BIC and ½ BIC were used to evaluate the kinetics of bacterial growth in the presence of antimicrobials (CMNP, FMNP and carvacrol solution). The results of this test showed that viable counts of Salmonella and L.monocytogenes were below the detection limit (1.69 log CFU/mL) after 2 h incubation (37 °C) using CMNP at the BIC. The wall material, rehydrated chia and flaxseed mucilages, reduced L. monocytogenes growth during 24 h. However, unloaded nanoparticles kept the viable counts of both microorganisms 2-5 log CFU/mL below the control curve of microbial growth during the 48 h experiment, suggesting that nanostructured mucilages potentiate antimicrobial properties. The results indicate that CMNP and FMNP have potential for use as food preservatives.

Characterization, Bioactivity and Application of Chitosan-Based Nanoparticles in a Food Emulsion Model.

In this study, chitosan nanoparticles (CNPs) were prepared by the ionic gelation technique with tripolyphosphate (TPP), and the effect of CNP composition and physicochemical characteristics were evaluated. After the synthesis optimization, CNPs were used as carriers for a fish protein hydrolysate (FPH) with bioactive properties (CNPH). The physicochemical characteristics, antioxidant capacity and antimicrobial, antihypertensive and emulsifier properties of unloaded and loaded CNPs in a food system model were studied. CNPH showed a uniform particle distribution, size ~200 nm, high stability (zeta potential around 30 mV), radical scavenging activity and increased antimicrobial activity against Staphylococcus aureus, Shigella sonnei and Aeromonas hydrophila. Additionally, CNPH showed an angiotensin I-converting enzyme (ACE)-inhibitory activity of 63.6% and, when added to a food emulsion model, this system containing CNPs, with or without FHP, exhibited improved food emulsion stability. Thus, CNPs were able to carry the FPH while maintaining their bioactive properties and can be an alternative to the delivery of bioactive peptides with potential as an emulsion stabilizer for food applications.