Directly brined black table olives of Bosana variety are a traditional food product of Sardinia island (Italy), spontaneously fermented by yeasts among other microorganisms. However, as far as we know, the identification, biotechnological and probiotic potential of this yeast community has not been investigated yet. In this work, a total of 72 yeast isolates previously obtained from Bosana olive brines were first genotyped by Random Amplified Polymorphic DNA (RAPD-PCR) analysis with primer M13, and then identified by sequencing of D1/D2 domains of rDNA 26S gene. The dominant species were Wickerhamomyces anomalus and Nakazawaea molendini-olei, albeit Candida diddensiae, Candida boidinii, Zygotorulaspora mrakii, and Saccharomyces cerevisiae were also present in lower proportions. For the different biotypes of yeasts obtained, the multivariate analysis of their technological (esterase, lipase and β-glucosidase activities, growth in presence of oleuropein, resistance and susceptibility to NaCl) and probiotic (removal of cholesterol, gastric and pancreatic digestions, biofilms assays alone and in co-culture with Lactobacillus pentosus) features, showed that W. anomalus Wa1 exhibited the best technological characteristics, while S. cerevisiae Sc24 and C. boidinii Cb60 showed promising probiotic features. Therefore, they may have potential application as multifunctional starters, alone or in combination with lactic acid bacteria, during olive processing, albeit further studies are necessary to validate these results.

In the present study, a total of 554 lactic acid bacteria (LAB) isolates were obtained from the olive surface of Manzanilla, Gordal, and Aloreña cultivars processed as green Spanish-style or directly brined (natural) olives. The isolates obtained from industrial processes were genotyped by rep-PCR with primer GTG5, collecting a total of 79 different genotypes. The α-biodiversity indexes showed that the LAB diversity was higher in the biofilms on the fruits which followed the Spanish-style process than in those just brined. Sixteen genotypes had a frequency higher >1% and were identified, by multiplex PCR recA gene and 16S gene sequencing, as belonging to L. pentosus (n=13) and L. plantarum (n=3) species. A multivariate analysis based on a dataset with 89,744 cells, including technological (resistance to salt and pH, production of lactic acid, auto and co-aggregation with yeast species, β-glucosidase and esterase activities), and potential probiotic characteristics (survival to gastric and pancreatic digestions, resistance to antibiotics, inhibition of pathogens, presence of bsh genes, cholesterol removal, hemolytic, α-glucosidase, β-galactosidase, and phytase activities) showed that the 16 genotypes could be grouped into 3 great phenotypes. Thus, the genotype biodiversity in table olive biofilms was limited but, at phenotype level, it was even lower since L. pentosus predominated clearly (80.15% isolates). L. pentosus Lp13 was the genotype with the most promising characteristics for its use as a multifunctional starter, with this strain being and ubiquitous microorganism present in both natural and lye-treated olive fermentations.

In this work, Manzanilla Spanish-style green table olive fermentations were inoculated with Lactobacillus pentosus LPG1, Lactobacillus pentosus Lp13, Lactobacillus plantarum Lpl15, the yeast Wickerhanomyces anomalus Y12 and a mixed culture of all them. After fermentation (65 days), their volatile profiles in brines were determined by gas chromatography-mass spectrometry analysis. A total of 131 volatile compounds were found, but only 71 showed statistical differences between at least, two fermentation processes. The major chemical groups were alcohols (32), ketones (14), aldehydes (nine), and volatile phenols (nine). Results showed that inoculation with Lactobacillus strains, especially L. pentosus Lp13, reduced the formation of volatile compounds. On the contrary, inoculation with W. anomalus Y12 increased their concentrations with respect to the spontaneous process, mainly of 1-butanol, 2-phenylethyl acetate, ethanol, and 2-methyl-1-butanol. Furthermore, biplot and biclustering analyses segregated fermentations inoculated with Lp13 and Y12 from the rest of the processes. The use of sequential lactic acid bacteria and yeasts inocula, or their mixture, in Spanish-style green table olive fermentation could be advisable practice for producing differentiated and high-quality products with improved aromatic profile.

In this work, a total of 72 samples of non-thermally treated commercial table olives were obtained from different markets of the world. Then, prokaryotic diversity in olive biofilms was investigated by metataxonomic analysis. A total of 660 different OTUs were obtained, belonging to Archaea (2.12%) and Bacteria domains (97.88%). From these, 41 OTUs with a proportion of sequences ≥ 0.01% were studied by compositional data analysis. Only two genera were found in all samples,Lactobacillus, which was the predominant bacteria in the biofilm consortium (median 54.99%), andPediococcus(26.09%).Celerinatantimonas,Leuconostoc, Alkalibacterium, Pseudomonas, Marinilactibacillus,Weissella,and the familyEnterobacteriaceaewere also present in at least 80% of samples. Regarding foodborne pathogens, onlyEnterobacteriaceae, Vibrio,andStaphylococcuswere detected in at least 91.66%, 75.00%, and 54.10% of samples, respectively, but their median values were always below 0.15%. Compositional data analysis allowed discriminating between lye treated and natural olive samples, as well as between olives packaged in glass, PET and plastic bags.Leuconostoc, Celerinatantimonas,andAlkalibacteriumwere the bacteria genera with a higher discriminant power among samples. These results expand our knowledge of the bacteria diversity in olive biofilms, providing information about the sanitary and hygienic status of this ready-to-eat fermented vegetable.