The objective of this work was to characterize the microbiota of breast milk in healthy Spanish mothers and to investigate the effects of lactation time on its diversity. A total of ninety-nine human milk samples were collected from healthy Spanish women and were assessed by means of next-generation sequencing of 16S rRNA amplicons and by qPCR. Firmicutes was the most abundant phylum, followed by Bacteroidetes, Actinobacteria, and Proteobacteria. Accordingly, Streptococcus was the most abundant genus. Lactation time showed a strong influence in milk microbiota, positively correlating with Actinobacteria and Bacteroidetes, while Firmicutes was relatively constant over lactation. 16S rRNA amplicon sequencing showed that the highest alpha-diversity was found in samples of prolonged lactation, along with wider differences between individuals. As for milk nutrients, calcium, magnesium, and selenium levels were potentially associated with Streptococcus and Staphylococcus abundance. Additionally, Proteobacteria was positively correlated with docosahexaenoic acid (DHA) levels in breast milk, and Staphylococcus with conjugated linoleic acid. Conversely, Streptococcus and trans-palmitoleic acid showed a negative association. Other factors such as maternal body mass index or diet also showed an influence on the structure of these microbial communities. Overall, human milk in Spanish mothers appeared to be a complex niche shaped by host factors and by its own nutrients, increasing in diversity over time.

Salmonella is a major food-borne pathogen around the world. In the European Union (EU), this pathogen is responsible of more than 90,000 human cases of salmonellosis every year. Salmonellosis in normally linked to the consumption of contaminated food, especially poultry products as meat, eggs and the products elaborated with them. Several control measures have been implemented in the EU to reduce the prevalence of Salmonella in the food chain. However, the ability of Salmonella to form biofilm along the food chain difficult its eradication. Also, ineffective cleaning and disinfection measures favors biofilm formation. The widespread use of biocides along the food chain has led to the emergence of resistant Salmonella strains. Therefore, it is necessary to look for alternatives to biocides to eradicate Salmonella biofilms. In this chapter we evaluate the use of bacteriocins and bacteriophages and their derivatives as a new alternative to eliminate Salmonella biofilms along the food chain.

Antimicrobial and biocide resistance is a major public health problem today. Therefore, one of the main scientific challenges nowadays is the search for alternatives to these substances. One of these potential alternatives are the bacteriocins. Microbiota are a potential source of bacteriocin-producing bacteria that need to be studied. In this study, a total of 40 samples of human milk and 10 samples of cow milk were collected from healthy individuals and stored at −20 °C until use. Colonies isolated from these samples that showed antimicrobial activity against Lactobacillus delbrueckii ssp. bulgaricus in the overlaid assays were selected. Well diffusion assays were carried out with the cell-free supernatant (CFS) from these colonies neutralized to pH and inhibition zones were recorded. The activity against eight common bacterial pathogens was evaluated. A total of 32 colonies with potential antimicrobial activity were isolated. The neutralized CFS of 10 strains showed antimicrobial activity against at least one pathogen tested in the well diffusion assays. Eight of the 10 CFS inhibited the growth of Staphylococcus aureus. These CFS also showed activity against Staphylococcus epidermidis, Streptococcus agalactiae, Pseudomonas aeruginosa , L. monocytogenes, Clostridium perfringens and Clostridioides difficile. The next steps of the research will be 16s rRNA sequencing to identify the species of isolates and mass spectrometry to determine the antimicrobial product produced by isolation. Finally, this study demonstrated that milk microbiota are a potential source of new producing bacteriocin bacteria that can be used in the formulation of new food products.