The properties of probiotics such as lactic acid bacteria (LAB) have been widely studied over the last decades. In the present study, four different LAB species, namely Lactobacillus gasseri ATCC 33323, Lacticaseibacillus rhamnosus GG ATCC 53103, Levilactobacillus brevis ATCC 8287, and Lactiplantibacillus plantarum ATCC 14917, were investigated in order to determine their ability to survive in the human gut. They were evaluated based on their tolerance to acids, resistance to simulated gastrointestinal conditions, antibiotic resistance, and the identification of genes encoding bacteriocin production. All four tested strains demonstrated high resistance to simulated gastric juice after 3 h, and the viable counts revealed declines in cell concentrations of less than 1 log cycle. L. plantarum showed the highest level of survival in the human gut, with counts of 7.09 log CFU/mL. For the species L. rhamnosus and L. brevis, the values were 6.97 and 6.52, respectively. L. gasseri, after 12 h, showed a 3.96 log cycle drop in viable counts. None of the evaluated strains inhibited resistance to ampicillin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline, or chloramphenicol. With regard to bacteriocin genes, the Pediocin PA gene was identified in Lactiplantibacillus plantarum ATCC 14917, Lacticaseibacillus rhamnosus GG ATCC 53103, and Lactobacillus gasseri ATCC 33323. The PlnEF gene was detected in Lactiplantibacillus plantarum ATCC 14917 and Lacticaseibacillus rhamnosus GG ATCC 53103. The Brevicin 174A and PlnA genes were not detected in any bacteria. Moreover, the potential antioxidant activity of LAB’s metabolites was evaluated. At the same time, the possible antioxidant activity of metabolites of LAB was first tested using the free radical DDPH^• (a, a-Diphenyl-β-Picrylhydrazyl) and then evaluated with regard to their radical scavenging activity and inhibition against peroxyl radical induced DNA scission. All strains showed antioxidant activity; however, the best antioxidant activity was achieved by L. brevis (94.47%) and L. gasseri (91.29%) at 210 min. This study provides a comprehensive approach to the action of these LAB and their use in the food industry.

Background: The identification of bacterial species in fermented PDO (protected designation of origin) cheese is important since they contribute significantly to the final organoleptic properties, the ripening process, the shelf life, the safety and the overall quality of cheese. Methods: Ten commercial PDO feta cheeses from two geographic regions of Greece, Epirus and Thessaly, were analyzed by 16S metagenomic analysis. Results: The biodiversity of all the tested feta cheese samples consisted of five phyla, 17 families, 38 genera and 59 bacterial species. The dominant phylum identified was Firmicutes (49% of the species), followed by Proteobacteria (39% of the species), Bacteroidetes (7% of the species), Actinobacteria (4% of the species) and Tenericutes (1% of the species). Streptococcaceae and Lactobacillaceae were the most abundant families, in which starter cultures of lactic acid bacteria (LAB) belonged, but also 21 nonstarter lactic acid bacteria (NSLAB) were identified. Both geographical areas showed a distinctive microbiota fingerprint, which was ultimately overlapped by the application of starter cultures. In the rare biosphere of the feta cheese, Zobellella taiwanensis and Vibrio diazotrophicus, two Gram-negative bacteria which were not previously reported in dairy samples, were identified. Conclusions: The application of high-throughput DNA sequencing may provide a detailed microbial profile of commercial feta cheese produced with pasteurized milk.

This e research focused on the detection and identification of genetic polymorphisms in exon 7 of the β-casein CSN2 gene in blood samples from Greek Holstein cows and from local breeds of cattle, such as Vrachykeratiki, Katerinis, and Sykias. For this purpose, DNA was isolated from 780 blood samples obtained from Greek Holstein cows, 86 from three local breeds of cattle, namely Brachyceros, Katerinis, and Sykias, and 14 from Greek buffalo. The desired region of exon 7 was amplified by PCR, resulting in 121 and 251 bp products in bovine and buffalo samples. The PCR product was digested with restriction fragment length polymorphism (RFLP) on agarose gels. The restriction enzymes DdeI and TaqI were used. All of the blood samples had the amplified size. The results showed that 74.4% of the Greek Holstein cows had the A2A2 β-casein genotype, the three native breads Vrachykeratiki had 57.7%, and the other two had 100% of the A2A2 β-casein. From the 14 Greek buffalo, 100% had the A2A2 β-casein.

Lactic acid bacteria (LAB) play an important role as natural food preservatives. However, the characterization of the variety of their metabolites is limited. The objective of this study was to determine the production of specific metabolites of Lacticaseibacillus rhamnosus, Levilactobacillus brevis and Lactiplantibacillus plantarum by an optimized liquid chromatography with an ultraviolet/diode detection (HPLC-UV/DAD) method and to investigate their potential antimicrobial activity against specific food pathogens. Based on the results of this study, the main metabolites detected in Levilactobacillus brevis were 103.4 μg mL⁻¹ DL-p-Hydroxyphenyllactic acid (OH-PLA) and 2.59 μg mL⁻¹ vanillic acid, while 216.2 μg mL⁻¹ OH-PLA, 19.0 μg mL⁻¹ salicylic acid, 3.7 μg mL⁻¹ vanillic acid, 6.9 μg mL⁻¹ ferulic acid, 4.2 μg mL⁻¹ benzoic acid and 1.4 μg mL⁻¹ 4-Hydrocinnamic acid were identified in the Lactiplantibacillus plantarum strain and 147.6 μg mL⁻¹ OH-PLA and 4.9 μg mL⁻¹ ferulic acid were identified in Lacticaseibacillus rhamnosus. This study provides alternative approaches for the molecules involved in the antimicrobial activity of food microorganism fermentation. These molecules may be used as antimicrobial ingredients in the food industry instead of conventional chemical preservatives.

Fermented foods such as table olives are produced through a spontaneous process that has been improved over the years, ensuring the safety and quality of the final product. The aim of the present work was to study the action of starter cultures of lactic acid bacteria (Lacticaseibacillus rhamnosus GG ATCC53103, Levilactobacillus brevis ATCC8287, and Lactiplantibacillus plantarum ATCC14917) which were previously shown to have probiotic and antioxidant potential during the fermentation of natural Greek-style black olives (Kalamata) in brine containing 6% (w/v) NaCl at a temperature of 20 °C for a period of 150 days. At a molecular level, the main metabolites in every fermentation process were identified using an HPLC method. The results showed that the concentration of the metabolites increased gradually, developing a stable pattern after the 90th day of fermentation. In addition, the DL-p-hydroxyphenyllactic acid (OH-PLA) was identified as the phenolic acid with the highest concentration, independently of the selected starter culture. Microbial genomic DNA was also extracted from the olives’ surface at the final stages of fermentation (150 days) and was subjected to 16S rRNA sequencing using the Nanopore MinION™ NGS tool, enabling a comprehensive analysis of the microbial community. According to the findings, the most abundant genera were Lactobacillus and Leuconostoc. To the best of our knowledge, this is the first study exploring these particular starters for olive fermentation.

Breast milk has been reported as a bacteria source that affects infant gut microbiota development. The present study utilizes a realtime PCR method to identify Lactobacillus and Bifidobacterium spp. in the breast milk of healthy women and attempts to identify factors affecting those human milk bacteria. Breast milk samples—both colostrum and mature milk—of 100 healthy women, were collected in Greece along with data about the demographic factors and nutritional habits of the volunteers. The colostrum samples were found to have higher percentages of either Bifidobacterium or Lactobacillus (76.9% and 48.6%, respectively) compared to the mature milk samples. For younger women, aged from 18 to 29 years, and women from rural areas, bacteria were detected in higher incidence than for older groups and women in urban areas, respectively. Moreover, for high-BMI women, bacteria were detected in lower incidence than for those with normal BMI. Probiotic supplements did not affect the composition of the breast milk-identified bacteria. Various factors such as lactation stage, maternal age, maternal weight, and residential location may contribute to the presence of those species in human milk. RT PCR has significant potential for the microbiological analysis of human milk.