This study aimed to define a consortium of lactic acid bacteria (LAB) that will bring added value to dried fresh cheese through specific probiotic properties and the synthesis of bioactive peptides (biopeptides). The designed LAB consortium consisted of three Lactobacillus strains: S-layer carrying Levilactobacillus brevis D6, exopolysaccharides producing Limosilactobacillus fermentum D12 and plantaricin expressing Lactiplantibacillus plantarum D13, and one Enterococcus strain, Enterococcus faecium ZGZA7-10. Chosen autochthonous LAB strains exhibited efficient adherence to the Caco-2 cell line and impacted faecal microbiota biodiversity. The cheese produced by the LAB consortium showed better physicochemical, textural and sensory properties than the cheese produced by a commercial starter culture. Liquid chromatography coupled with matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry (LC-MALDI-TOF/TOF) showed the presence of 18 specific biopeptides in dried fresh cheeses. Their identification and relative quantification was confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using multiple reaction monitoring (MRM). The results also showed that their synthesis resulted mainly from β-casein and also α-S1 casein degradation by proteolytic activities of the LAB consortium. The designed LAB consortium enhanced the functional value of the final product through impact on biopeptide concentrations and specific probiotic properties.

Human milk not only provides a perfect balance of nutrients to meet all the needs of the infant in the first months of life but also contains a variety of bacteria that play a key role in tailoring the neonatal faecal microbiome. Microbiome analysis of human milk and infant faeces from mother-breastfed infant pairs was performed by sequencing the V1–V3 region of the 16S rRNA gene using the Illumina MiSeq platform. According to the results, there is a connection in the composition of the microbiome in each mother-breastfed infant pair, supporting the hypothesis that the infant’s gut is colonised with bacteria from human milk. MiSeq sequencing also revealed high biodiversity of the human milk microbiome and the infant faecal microbiome, whose composition changes during lactation and infant development, respectively. A total of 28 genetically distinct strains were selected by hierarchical cluster analysis of RAPD-PCR (Random Amplified Polymorphic DNA-Polymerase Chain Reaction) electrophoresis profiles of 100 strains isolated from human milk and identified by 16S RNA sequencing. Since certain cellular molecules may support their use as probiotics, the next focus was to detect (S)-layer proteins, bacteriocins and exopolysaccharides (EPSs) that have potential as therapeutic biomolecules. SDS-PAGE (Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis) coupled with LC-MS (liquid chromatography-mass spectrometry) analysis revealed that four Levilactobacillus brevis strains expressed S-layer proteins, which were identified for the first time in strains isolated from human milk. The potential biosynthesis of plantaricin was detected in six Lactiplantibacillus plantarum strains by PCR analysis and in vitro antibacterial studies. 1H NMR (Proton Nuclear Magnetic Resonance) analysis confirmed EPS production in only one strain, Limosilactobacillus fermentum MC1. The overall microbiome analysis suggests that human milk contributes to the establishment of the intestinal microbiota of infants. In addition, it is a promising source of novel Lactobacillus strains expressing specific functional biomolecules.

Research background. Increasing awareness of the importance of nutrition in health promotion and disease prevention has driven to the development of foods for special medical purposes (FSMPs). In this research, the probiotic strain Lacticaseibacillus paracasei subsp. paracasei (Lacticaseibacillus casei 431®) was incorporated into FSMPs to develop an innovative product. The aim was to investigate the influence of FSMPs matrix on the specific probiotic properties of L. casei 431® in vitro. Experimental approach. A series of in vitro experiments were performed as part of the probiotic approach. After evaluation of antibiotic susceptibility profiles, functional properties such as survival under simulated gastrointestinal tract (GIT) conditions, bile salt deconjugation activities, cholesterol assimilation, antagonistic activity against spoilage bacteria, and adhesion to Caco-2 cell line monolayers and extracellular matrix proteins were investigated. Results and conclusions. L. casei 431® strain, both the lyophilised one and the one isolated from FSMPs matrix, effectively survived the adverse simulated gastrointestinal conditions without significant effect of the food matrix. The effect of FSMPs matrix on the deconjugation activity of bile salts of L. casei 431® was minimal, however, cholesterol assimilation was increased by 16.4 %. L. casei 431® showed antibacterial activity against related lactic acid bacteria whether it was implemented in FSMPs or not. Conversely, the probiotic strain isolated from FSMPs matrix showed significantly higher inhibitory activity against 6 potential pathogens than the lyophilised culture. The autoaggregation ability of L. casei 431® cells was not affected by FSMPs matrix. The adhesion of L. casei 431® bacterial cells to the extracellular matrix proteins was reduced after treatment with proteinase K, with the highest adhesion observed to laminin. Adhesion of L. casei 431® reduced E. coli 3014 binding by 1.81 log units and S. Typhimurium FP1 binding by 1.85 log units to Caco-2 cell lines, suggesting the potential for competitive exclusion of these pathogens. Novelty and scientific contribution. The results support the positive influence of FSMPs matrix on the specific probiotic properties of L. casei 431®, such as antibacterial activity, bile salts deconjugation and cholesterol assimilation, while the incorporation of this probiotic strain brought additional functional value to the FSMPs. The achieved synergistic effect of the join application of L. casei 431® and innovative FSMPs matrix contributed to the development of the novel formulation of an improved functional food product with added value.

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The paper shows results of the archaeological research conducted at the site Stari grad Ljubuški, in Ljubuški. Those excavations were of great significance because they were the first archaeological excavations of that remarkable Herzegovinian monument known among the people as "Kula hercega Stjepana". Conservation and restoration work followed the archaeological excavations. They will save the fortress from the collapse, since some parts of its walls and other constructive elements were almost destroyed or already collapsed. Archaeological research conducted by the