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The presence of surface (S)-layer proteins as a rare property of lactic acid bacteria plays an important role in the expression of the probiotic properties of the producer strain and protects the cells from unfavourable environmental conditions. Therefore, the aim of this work was to evaluate the role of S-layer proteins of four Levilactobacillus brevis MB1, MB2, MB13, and MB20 strains, previously isolated from human milk. The results showed better survival under simulated gastrointestinal tract (GIT) conditions and better adhesion to Caco-2 cells, achieved when S-proteins were present on the surface of the examined probiotic Levilactobacillus brevis strains. Free cells and cells microencapsulated in alginate, with or without galactooligosaccharides (GOS) and fructooligosaccharides (FOS), survived in high numbers during freeze-drying and one-month storage at 4°C, as well as during exposure to simulated GIT conditions (>108 CFU/g), thus fulfilling the primary technological criterion for the selection of probiotic strains. Higher survival of freeze-drying of encapsulated cells than free cells, can be attributed to the protective effect of alginate, GOS and FOS as prebiotic agents. However, GOS was more protective than FOS in maintaining the viability of microencapsulated Levilactobacillus brevis MB1, MB2, MB13 and MB20 cells in alginate during storage and under the harsh conditions of GIT and freeze-drying. [ABSTRACT FROM AUTHOR]

Proteases of Lactobacillus and Lactococcus strains catalyze casein degradation in fermented dairy products, which can result in the production of bioactive peptides. Proteolytic properties of a selection of lactic acid bacteria (LAB) strains previously isolated in Croatia, including Lactococcus lactis and Lactobacillus strains, are described. All strains of Lactobacillus and Lactococcus showed an Fmc+ phenotype that can be associated with efficient growth in milk. The significant caseinolytic effect, after incubation of culture supernatant or concentrated cell biomass, was observed for Levilactobacillus brevis D6 and Lactiplantilactobacillus plantarum D13 after growth in the optimal growth medium, while for Lactoccocus lactis ZGBP5-32 and Levilactobacillus brevis SF9B strains after growth in skimmed milk. To assess the LAB growth in skimmed milk, the acidification rate was monitored. Statistically, significant acidification capacity was determined for L. plantarum D13 in the optimal medium and by the proteolytic strain Lactobacillus helveticus M92 in skimmed milk. After extraction of proteinases from the strains with caseinolytic activity, protein samples were analysed by the SDS-PAGE. The protein extract of the Lc. lactis ZGBP5-32 and ZGZA7-10, retained proteolytic activity even at very low concentrations. The ultrafiltration improved protein extraction. The crude extract possibly contained putative protease, as a decrease in contaminating proteins was confirmed by SDSPAGE in samples of L. brevis D6 and SF9B, L. fermentum D12 and L. plantarum D13. [ABSTRACT FROM AUTHOR]

Fermented sausages have been manufactured traditionally in the Mediterranean basin. Istrian sausages are one of the many regionally specific products distinguished by the specific traditional recipes and consequentially exceptional sensory characteristics. The aim of this study was to evaluate sensory, physico-chemical and microbiological changes occurring during production and ripening of Istrian sausages made at traditional households, and to compare the results obtained with the identical product produced in industrial conditions. Additional goal was to characterise the autochthonous lactic acid bacteria populations and to obtain the candidate starter strains. Production stages and product ripening parameters could be strictly defined in controlled industrial conditions of the production process, and the results of this study can serve to standardise the production of traditional Istrian sausages. The results from sensory and physico-chemical examination do not show differences in relation to sausages of a similar type investigated in other papers. Lactic acid bacteria were favoured during fermentation at rising salt concentrations, consisting mostly of Lactobacillus strains followed with the abundance of coagulase-negative cocci. Further, we targeted the autochthonous lactic LAB population to select the candidate starter strains for controlled sausage fermentation. Fifty Gram-positive strains with catalase-negative phenotype, rod-shape or cocci morphology, effective acidification kinetics and production of lactic acid identified by sequence analysis of 16S rRNA were considered LAB. The selected LAB strains were able to grow in a range of pH values and high salt concentration important for their metabolic activity during sausage fermentation. According to our results, Lactobacillus sakei, which is a prevalent strain during fermentation of various types of fermented sausages, was frequently detected. Lactobacillus plantarum and Lactobacillus curvatus were also detected in abundance, implying their possible competitiveness among the complex microbial population. Isolated autochthonous strains L. curvatus S30, L. sakei S32 and L. plantarum S50 produced high lactic acid concentrations, tolerated NaCl concentrations present during sausage fermentation, survived at high concentrations of hydrogen ions and inhibited six pathogens associated with contamination of meat products in a strain-specific manner. [ABSTRACT FROM AUTHOR]

The antibiotic susceptibility and antimicrobial activity, as food safety parameters important for application of autochthonous lactic acid bacteria (LAB), that previously satisfied technological criteria for functional starter cultures in fresh cheese production were examined. Soluble whole cell protein patterns of autochthonous LAB strains from fresh cheese, obtained by SDS-PAGE, revealed the presence of two predominant strains, which were identified as Lactobacillus fermentum A8 and Enterococcus faecium A7. These strains were not resistant and shown susceptibility to antibiotics: ampicillin, bacitracin, penicillin G, azithromycin, chloramphenicol, clarithromycin, clindamycin, spiramycin, tetracycline, streptomycin, neomycin, gentamicin, erythromycin, rifampicin and novo-biocin. Lb. fermentum A8 strain displayed phenotypic resistance to vancomycin, but this resistance is intrinsic, not transferable and it is acceptable from the safety aspect. The capacity of Lb. fermentum A8 and Ec. faecium A7 to inhibit growth of test-microorganisms Listeria monocytogenes ATCC 11911, Escherichia coli 3014, Salmonella enterica serovar Typhimurium FP1 and Staphylococcus au-reus 3048, was also analysed. According to obtained results, Lb. fermentum A8 and Ec. faecium A7 are safe from the aspect of spreading antibiotic resistance and could be useful as bioprotective cultures that inhibit common bacterial food contaminants, including L. monocytogenes. [ABSTRACT FROM AUTHOR]

The use of commercial starter cultures in fresh cheese production from pasteurized milk results in the loss of typical characteristics of artisan fresh cheese due to the replacement of complex native microbiota with a defined starter culture. Hence, the aim of this research is to isolate and characterize dominant lactic acid bacteria (LAB) in artisan fresh cheese and to evaluate their capacity as autochthonous starter cultures for fresh cheese production. Fifteen most prevalent Gram-positive, catalase-negative and asporogenous bacterial strains were selected for a more detailed characterization. Eleven lactic acid bacterial strains were determined to be homofermentative cocci and four heterofermentative lactobacilli. Further phenotypic and genotypic analyses revealed that those were two different LAB strains with high acidifying and proteolytic activity, identified as Lactobacillus fermentum A8 and Enterococcus faecium A7. These two autochthonous strains, alone or in combination with commercial starter, were used to produce different types of fresh cheese, which were evaluated by a panel. Conventional culturing, isolation, identification and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) procedures, applied to the total fresh cheese DNA extracts, were employed to define and monitor the viability of the introduced LAB strains and their effect on the final product characteristics. Production of fresh cheese using a combination of commercial starter culture and selected autochthonous strains resulted in improved sensorial properties, which were more similar to the ones of spontaneously fermented fresh cheese than to those of cheese produced with only starter culture or selected strains. After 10 days of storage, that cheese retained the best sensorial properties in comparison with all other types of cheese. The presence of inoculated autochthonous and starter cultures and their identification was demonstrated by DGGE analysis. The obtained results indicate that autochthonous strains have a strong potential to enrich the flavour of industrially produced fresh type cheese under controlled conditions. [ABSTRACT FROM AUTHOR]

The antimicrobial activity of industrially important lactic acid bacteria as starter cultures and probiotic bacteria is the main subject of this review. This activity has been attributed to the production of metabolites such as organic acids (lactic and acetic acid), hydrogen peroxide, ethanol, diacetyl, acetaldehyde, acetoine, carbon dioxide, reuterin, reutericyclin and bacteriocins. The potential of using bacteriocins of lactic acid bacteria, primarily used as biopreservatives, represents a perspective, alternative antimicrobial strategy for continuously increasing problem with antibiotic resistance. Another strategy in resolving this problem is an application of probiotics for different gastrointestinal and urogenital infection therapies. [ABSTRACT FROM AUTHOR]

In this research the application of bacterial strains Lactobacillus acidophilus M92, Lactobacillus plantarum L4 and Enterococcus faecium L3 in the production of fermented probiotic beverages from milk permeate enriched with 10 % (v/v) whey retentate, was investigated. In the previous researches of probiotic concept, probiotic properties of these three strains of lactic acid bacteria have been defined. At the end of controlled fermentation, probiotic strains have produced 7.4 g/L lactic acid, pH was decreased to 4.7, and number of live cells was around 108 CFU/mL. Number of viable count of probiotic bacteria, which were identified with RAPD (RandomAmplifiedPolymorphicDNA)(Random Amplified Polymorphic DNA) method, was maintained at around 107 CFU/mL during 28 days of the preservation at 4 °C. Furthermore, a spontaneous fermentation of milk permeate enriched with 10 % (v/v) of whey retentate was carried out and lactic acid bacteria present in these substrates were isolated. All of these bacterial strains have rapidly acidified the growth media and have shown antibacterial activity against chosen test-microorganisms, what are important properties of potential starter cultures for the fermentation of dairy products. The results of biochemical API analysis have identified isolated strains as Lactococcus lactis subsp. lactis and Lactobacillus helveticus. [ABSTRACT FROM AUTHOR]

Lactic acid bacteria (LAB) produce a broad spectrum of exopolysaccharides (EPSs), commonly used as texturizers in food products. Due to their potential contribution to LAB probiotic properties, like adhesion to human epithelial cells and competitive exclusion of pathogens from human intestinal epithelial cells, this study was focussed on the structural and functional characterization of the EPSs produced by two Limosilactobacillus fermentum strains – MC1, originating from mother's milk, and D12, autochthonous from Croatian smoked fresh cheese. Whole-genome sequencing and functional annotation of both L. fermentum strains by RAST server revealed the genes involved in EPS production and transport, with some differences in functionally related genes. EPSs were extracted from the cell surface of both bacterial strains and purified by size-exclusion chromatography. Structural characterization of the EPSs, achieved by chemical analyses and 1D and 2D NMR spectroscopy, showed that both strains produce an identical mixture of three different EPSs containing galactofuranose and glucopyranose residues. However, a comparison of the functional properties showed that the MC1 strain adhered better to the Caco-2 cell line and exhibited stronger antimicrobial effect against Salmonella enterica serovar Typhimurium FP1 than the D12 strain, which may be attributed to the potential bacteriocin activity of the MC1 strain. [Display omitted] • two EPS-producing L. fermentum strains were isolated from different sources. • WGS showed that the genomes of L. fermentum MC1 and D12 are slightly different. • both strains produce an identical mixture of three different EPSs. • in vitro experiments showed different functional properties of the two strains. [ABSTRACT FROM AUTHOR]