Raw goat and ewe’s milk samples were used for the isolation of seven lactic acid bacteria new strains. After testing hemolytic activity and resistance to antibiotics, specific functional properties were evaluated; Lactococcus lactis subsp. lactis FBM_1321 and Lacticaseibacillus paracasei FBM_1327 strains resulted in the highest cholesterol assimilation percentages ranging from 28.78 to 30.56%. In addition, strong adhesion capacity to differentiated Caco-2 cells (1.77–21.04%) was mapped, and the lactobacilli strains exhibited strong antagonistic activity against foodborne pathogens compared to lactococci. The strains were able to grow at low pH and high NaCl concentrations, conditions that prevail in food systems (cell counts ranged from 1.77 to 8.48 log CFU/mL after exposure to pH 3 and from 5.66 to 9.52 log CFU/mL after exposure to NaCl concentrations up to 8%). As a next step, freeze-dried immobilized Lc. paracasei FBM_1327 cells on oat flakes were used for the preparation of functional yoghurt and ayran products. Cell loads of the functional strain remained high and stable in both products (7.69 log CFU/g in yoghurt and 8.56 log CFU/g in ayran after 30 days of storage at 4 °C) throughout their shelf life. No significant changes in the volatile profile were noticed, and the new products were accepted by the panel during the sensory evaluation.

Seven wild-type lactic acid bacteria, belonging to Lactiplantibacillus plantarum and Lactococcus cremoris species, were isolated from beetroots and white mushrooms and evaluated for their safety and functional profile. Lc. cremoris isolates were sensitive to all antibiotics tested, while L. plantarum strains exhibited resistance in certain antibiotics. Among them, Lc. cremoris FBMS_5810 showed the highest cholesterol removal ability (51.89%) and adhesion capacity to Caco-2 cell lines (32.14%), while all plant origin strains exhibited strong antagonistic and inhibitory activity against foodborne pathogens, as well as high survival potential during an in vitro digestion model. Subsequently, freeze-dried immobilized Lc. cremoris FBMS_5810 cells on oat flakes were prepared with initial cell loads >8.5 log CFU/g, and the effect of trehalose as a cryoprotectant in cell viability during storage at room and refrigerated temperatures for up to 180 days was studied. A significant reduction in cell loads was observed in all cases studied. However, freeze-dried immobilized Lc. cremoris FBMS_5810 cells on oat flakes prepared using trehalose as a cryoprotectant stored at 4 °C exhibited the highest cell viability (8.75 log CFU/g) after 180 days. In the next step, functional breakfast cereals enriched with freeze-dried immobilized Lc. cremoris FBMS_5810 cells on oat flakes (produced with (MLT) or without (ML) trehalose) were developed and stored at room and refrigerated temperatures for 180 days. The initial cell levels ≥ 9.18 log CFU/g were achieved, while a significant decrease was recorded during storage in all cases. The maintenance of cell loads ≥ 7.75 log CFU/g was documented in the case of both ML and MLT samples stored at 4 °C; however, the presence of trehalose in MLT samples resulted in cell viability 7.52 log CFU/g after 180 days of storage at room temperature. Importantly, the functional breakfast cereals were accepted by the panel during the sensory evaluation.

Background and Objectives: The development of non-dairy probiotic products is a challenge for the food industry, while cereals, as probiotic carriers, provide the means to incorporate probiotics, prebiotics, and fiber into the human diet. The present study investigated the effects of Lactococcus cremoris spp. immobilized on oat flakes on blood and urine biomarkers in a randomized placebo-controlled single-blind clinical trial. Materials and Methods: Fifty-four eligible participants were randomized into a placebo or probiotic group that consumed 5 g of oat flakes daily for 12 weeks. Blood and urine samples were collected at the baseline, 6 weeks, and 12 weeks to assess the glycemic, lipemic, inflammatory, immunological, and antioxidant biomarkers, as well as the vitamin levels. Results: The intervention group exhibited a significant reduction in their hs-CRP levels (p = 0.002) and a trend toward decreased IL-6 levels (p = 0.035) at week 12 compared to the control group, suggesting a potential anti-inflammatory effect. Additionally, a significant reduction in insulin levels was observed in the probiotic group at week 6, with a clinical trend toward differentiation despite the absence of statistically significant differences between the groups. Furthermore, there were promising results regarding certain biomarkers, such as vitamin B12 and cortisol levels, in the probiotic group. Conclusions: The twelve-week consumption of Lactococcus cremoris spp. immobilized on oat flakes resulted in improvements in inflammatory, metabolic, and stress-related biomarkers. These results support the examined concept of non-dairy probiotic products, though further research is needed to confirm their efficacy and clarify their underlying mechanisms.

Nowadays, functional foods supplemented with health-promoting microorganisms have attracted consumer attention due to their health benefits. However, maintaining high cell loads, which consist of an essential requirement for conferring the health effect, is a real bottleneck for the food industry due to viability declines during food processing and storage. Hence, freeze-drying and cell immobilization have been suggested to enhance cell viability. The aim of our study was to assess the effect of freeze-dried immobilized P. acidilactici ORE5 on pistachio nuts on the functional regulation of the Katiki Domokou-type cheese microbiome. Supplementation of Katiki Domokou-type cheese with free or immobilized P. acidilactici ORE5 culture resulted in cell loads > 8.5 logcfu/g up to 7 days of storage. Both free and immobilized P. acidilactici ORE5 cells suppressed the growth of L. monocytogenes after deliberate inoculation, acting as a protecting shield. HS-SPME GC/MS analysis showed that the incorporation of P. acidilactici ORE5 culture in cheese resulted in an improved volatile compounds profile, as verified by the preliminary sensory evaluation. According to Next-Generation Sequencing analysis, a wide range of bacterial diversity was revealed among samples. The most abundant genus was Lactococcus in all samples, while the results showed an increased presence of Pediococcus spp. in cheese fortified with P. acidilactici ORE5 culture, highlighting the ability of the strain to survive in the final product. Furthermore, the incorporation of P. acidilactici ORE5 culture in cheese had a significant impact on cheese microbiome composition, as the presence of spoilage bacteria, such as Chryseobacterium, Acinetobacter and Pseudomonas, was significantly less compared to the control cheese, indicating quality improvement and prolongation of the product’s shelf-life.

Nowadays, functional foods supplemented with health-promoting microorganisms have attracted the consumers attention due to their health benefits. However, maintaining high cell loads, which consists of an essential requirement for conferring the health effect is a real bottleneck for the food industry, due to viability decline during food processing and storage. Hence, freeze-drying and cell immobilization have been suggested to enhance cell viability. The aim of our study was to assess the effect of freeze-dried immobilized P. acidilactici ORE5 on pistachio nuts on functional regulation of Katiki Domokou-type cheese microbiome. Supplementation of Katiki Domokou-type cheese with free or immobilized P. acidilactici ORE5 culture resulted in cell loads > 8.5 logcfu/g up to 7 days of storage. Both free and immobilized P. acidilactici ORE5 cells suppressed the growth of L. monocytogenes after deliberate inoculation, acting as a protecting shield. SPME GC/MS analysis showed that incorporation of P. acidilactici ORE5 culture in cheese resulted in an improved volatile compounds profile, as verified by the preliminary sensory evaluation. According to Next-Generation Sequencing analysis, a wide range of bacterial diversity was revealed among samples. The most abundant genus was Lactococcus in all samples, while the results showed increased presence of Pediococcus spp. in cheese fortified with P. acidilactici ORE5 culture, highlighting the ability of the strain to survive in the final product. Furthermore, the incorporation of P. acidilactici ORE5 culture in cheese had a significant impact on cheese microbiome composition, as the presence of spoilage bacteria, such as Chryseobacterium, Acinetobacter and Pseudomonas, were significantly lower compared to the control cheese, indicating quality improvement and prolongation of the product’s shelf- life.

The present study aimed to investigate the effect of a dietary intervention including free or immobilized cells of the presumptive probiotic Pediococcus acidilactici ORE5 on Corinthian currants, a food with beneficial impact in the condition of Type-1 Diabetes Mellitus (T1DM), on the microbiome composition of STZ-induced diabetic rats. Twenty four male Wistar rats were divided into four groups (n = 6 per group): healthy animals, which received the free (H_FP) or the immobilized Pediococcus acidilactici ORE5 cells (H_IPC), and diabetic animals, which received the free (D_FP) or the immobilized Pediococcus acidilactici ORE5 cells(D_IPC) for 4 weeks (10⁹ cfu/day, in all groups). At the end of the dietary intervention, the D_IPC group exerted a lower concentration of the inflammatory cytokine IL-1 beta compared to D_FP. Consumption of immobilized P. acidilactici ORE5 cells on Corinthian currants by diabetic animals led to increased loads of fecal lactobacilli and lower Enterobacteriaceae, coliforms, and Escherichia coli levels, while Actinobacteria phylum, Akkermansia, and Bifidobacterium genera abundances were increased, and fecal lactic acid was elevated. Overall, the results of the present research demonstrated that functional ingredients could ameliorate gut dysbiosis present in T1DM and could be used to design dietary patterns aiming at T1DM management. However, well-designed clinical trials are necessary, in order to confirm the beneficial effects in humans.