Effect of the cleaning and disinfection methods on the hygienic conditions of fermentation tanks of table olives (Olea europaea L.) Negrinha de Freixo cultivar.

This study aimed to evaluate and identify the microbial community attached to the surfaces of fermenter tanks used in table olive Negrinha de Freixo cultivar processing through molecular analysis and verify if the cleaning/disinfection was done correctly. Four fermentation tanks previously used in table olive processing were sampled at three different inside areas: upper, middle, and lower. Before sampling, four cleaning/disinfection methods were applied to the tanks, including (i) pressurised water; (ii) a disinfectant product used to clean bowls (Vasiloxe); (iii) 10% sodium hydroxide solution (caustic soda liquid); and (iv) a disinfectant product used by the wine industry (Hosbit). For each sample collected, mesophilic aerobic bacteria, yeast and moulds (YMC), lactic acid bacteria (LAB), as well as total coliforms (TC) and Pseudomonas aeruginosa were evaluated. The results showed significant differences between the different cleaning/disinfection methods applied. The fermenter sanitised with only pressurised water showed a greater abundance of microorganisms than the others. Mesophilic aerobic bacteria were the predominant population, with counts ranging between 2.63 and 5.56 log10 CFU/100 cm2, followed by the moulds (3.11-5.03 log10 CFU/100 cm2) and yeasts (2.42-5.12 log10 CFU/100 cm2). High diversity of microbial communities was observed between the different fermenter tanks. The most abundant species belonged to Aureobasidium, Bacillaceae, Cladosporium, and Rhodotorula genera. LAB, TC, and P. aeruginosa were not detected. This study hopes to improve hygienic conditions and increase the quality assurance and safety of the final product.

Enhancing the quality and safety of Nocellara del Belice green table olives produced using the Castelvetrano method.

The Castelvetrano method is the most widely used among the various table olive processing styles in Sicily. After debittering, the product is stored at low temperatures to prevent the growth of undesirable microorganisms. In an effort to enhance the production process, yeast isolates underwent genotypic characterization and technological screening. The screening process identified two yeast strains Candida norvegica OC10 and Candida boidinii LC1, which can grow at low temperatures and tolerate high pH values (up to 10) and salinity [10% (w/v)]. During the monitoring period, the inoculated trials showed limited presence of spoilage/pathogenic microorganisms. Additionally, the yeasts limited oxidative phenomena and softening of the drupes. The organic compounds detected were higher in the inoculated trials than in the control, and cold storage induced aromatic decay, which was less pronounced in the trial inoculated with C. norvegica. Sensory analysis revealed that the inoculated trials scored higher in sweetness, hardness and crispness.

A New Culture Medium Rich in Phenols Used for Screening Bitter Degrading Strains of Lactic Acid Bacteria to Employ in Table Olive Production.

The olive oil industry recently introduced a novel multi-phase decanter with the "Leopard DMF" series, which gives a by-product called pâté, made up of pulp and olive wastewater with a high content of phenolic substances and without pits. This study aims to create a new culture medium, the Olive Juice Broth (OJB), from DMF pâté, and apply it to select bacteria strains able to survive and degrade the bitter substances normally present in the olive fruit. Thirty-five different bacterial strains of Lactiplantibacillus plantarum from the CREA-IT.PE Collection of Microorganisms were tested. Seven strains characterized by ≥50% growth in OJB (B31, B137, B28, B39, B124, B130, and B51) showed a degradation of the total phenolic content of OJB ≥ 30%. From this set, L. plantarum B51 strain was selected as a starter for table olive production vs. spontaneous fermentation. The selected inoculant effectively reduced the debittering time compared to spontaneous fermentation. Hydroxytyrosol, derived from oleuropein and verbascoside degradation, and tyrosol, derived from ligstroside degradation, were produced faster than during spontaneous fermentation. The OJB medium is confirmed to be useful in selecting bacterial strains resistant to the complex phenolic environment of the olive fruit.

Fermentation starters and bacteriocins as biocontrol strategies for table olives preservation: a mini-review.

Biopreservation is a powerful strategy to prolong the shelf life of food products by applying naturally occurring microorganisms and/or their metabolites. Current food trends emphasise the need to develop alternatives for chemical or thermal preservation methods. In this line, different fermentation starters from table olives present the potential to control spoilage or pathogen-occurring microorganism in table olives storage. One of the most interesting family used as biopreservative culture is Lactobacillaceae and it has also been used in combination with yeasts as olive fermentation starter. Lactic acid bacteria, from Lactobacillaceae family, are characterised by the production of bacteriocins, proteins with the potential for preserving food by changing the organisation of the membrane of spoilage microorganisms. These bacteriocins-producing bacteria can be directly inoculated, although nanosystem technology is the most promising incorporation strategy. In table olives, the most commonly used starters are Lactiplantibacillus plantarum, Lactiplantibacillus pentosus, Saccharomyces cerevisiae, Wickerhamomyces anomalus, among others. These strains with biopreservation characteristics, inoculated alone or in mixed cultures, ensure food safety by conferring the product added value and prolonging product shelf life. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Microbial community and volatilome changes in brines along the spontaneous fermentation of Spanish-style and natural-style green table olives (Manzanilla cultivar).

Microbial community and volatilome of brines were monitored during the spontaneous fermentations of Spanish-style and Natural-style green table olives from Manzanilla cultivar. Fermentation of olives in the Spanish style was carried out by lactic acid bacteria (LAB) and yeasts, whereas halophilic Gram-negative bacteria and archaea, along with yeasts, drove the fermentation in the Natural style. Clear differences between the two olive fermentations regarding physicochemical and biochemical features were found. Lactobacillus, Pichia, and Saccharomyces were the dominant microbial communities in the Spanish style, whereas Allidiomarina, Halomonas, Saccharomyces, Pichia, and Nakazawaea predominated in the Natural style. Numerous qualitative and quantitative differences in individual volatiles between both fermentations were found. The final products mainly differed in total amounts of volatile acids and carbonyl compounds. In addition, in each olive style, strong positive correlations were found between the dominant microbial communities and various volatile compounds, some of them previously reported as aroma-active compounds in table olives. The findings from this study provide a better understanding of each fermentation process and may help the development of controlled fermentations using starter cultures of bacteria and/or yeasts for the production of high-quality green table olives from Manzanilla cultivar.

Delving into the study of lactic acid bacteria and yeasts distribution in table olive biofilms using a non-destructive procedure.

To turn table olives into appropriate carriers of beneficial bacteria and yeasts to consumers, it is essential to have reliable methods for analysing microorganisms in biofilms. This work validates the application of a non-destructive procedure to study the lactic acid bacteria and yeasts distribution in fruits during Spanish-style green table olive fermentations. Laboratory-scale fermentations were inoculated simultaneously with three Lactiplantibacillus pentosus strains (LPG1, 119, and 13B4) and two yeasts (Wickerhamomyces anomalus Y12 and Saccharomyces cerevisiae Y30), all of them natives of table olive fermentations. Data showed that L. pentosus LPG1 and yeasts W. anomalus Y12 were quite prone to colonise olive biofilms, but only the Lactiplantibacillus strain also can penetrate the epidermis of the fruit and colonise the flesh. Applying a non-destructive treatment consisting in shelling the fruits with glass beads led to obtaining similar lactic acid bacteria and yeast recovery than the classical stomacher destructive method. However, the glass bead procedure improved the quality of the metagenomics analysis (especially when using 16 S rRNA gene-based sequencing). Results show the great utility of procedures that do not destroy the fruit for studying fermented vegetable biofilms.

Comparative Study on Chemical Composition of Green and Black Table Olives Brines of the Endemic ''Sigoise'' Cultivar: Recovery of high-Added Values Compounds.

The effluents derived from the processing of table olives stand for a serious environmental problem. The study aims to valorize the brine water of table olives at different stages of ripening (green and black) of the Algerian variety Sigoise of Bejaia (East) and Mascara (West). The physico-chemical characterization revealed that these samples display have a high acid pH and salinity. The comparative study of phenolic levels exhibited showed very significant differences between the brine waters of green olives from Bejaia and Mascara, while the brines of black olives presented showed comparable levels. A high strong antioxidant potential was confirmed by DPPH (CI50 =0.35 µg/100 ml-0.50 µg/100 ml) and FRAP (CI50 =626.89 µg/100 ml-875.54 µg/100 ml) tests. Chemical screening by HPLC-DAD of the four samples identified high concentrations of hydroxytyrosol (HT) (390.4 mg/100 ml-360.8 mg/100 ml) and tyrosol (202.2 mg/100 ml-101.4 mg/100 ml). This study provided a deeper insight into the phenolic profile and the antioxidant potential of these brines.

The Fate of the Chlorophyll Derivatives in Olives Preserved and/or Packaged in Presence of Exogenous Copper.

Chlorophyll pigments are thought to be responsible for the highly appreciated green color of unfermented Castelvetrano-style table olives, but no studies have considered the effects of a controlled addition of copper during storage or packaging at the industrial level. For this purpose, chlorophyll derivatives were analyzed in Nocellara cultivar table olives debittered industrially using the Castelvetrano method, via means of HPLC and MS analyses, following the addition of copper in alkaline brines stored at 4 °C for 3 months in 220 L barrels, and during the subsequent storage in acid brines in commercial 400 g packages at 4 °C for up to 18 months. The presence of copper in storage or in packaging brines both contributed significantly to maintaining the green color of the olives, which was associated with a specific pattern of chlorophyll derivatives, as evidenced by principal component analysis. Notably, re-greening was rapidly achievable also for olives that had yellowed for 18 months at a copper concentration below the limit of EU legislation. Finally, by means of PCA, we also demonstrated that a short-term thermic treatment can work as an accelerated predictive tool in determining the fate of chlorophyll derivatives.

The Fate of Chlorophylls in Alkali-Treated Green Table Olives: A Review.

This paper reviews the current knowledge regarding modifications to chlorophylls during the processing of green table olives treated with alkali. Particular attention is given to the pheophytinization reactions (substitution of Mg2+ by 2H+ in the chlorophyll chromophore group) that can take place because of pH and/or temperature changes and the possible sequential substitution of the 2H+ with Cu2+ within the chlorophyll porphyrin ring. These reactions may have a direct impact on the commercial value of olive productions as some naturally forming Cu-chlorophylls complexes (i) are identical to strictly forbidden colorants for table olives (E141) and (ii) have been identified as responsible for the unwelcome appearance of the so-called green staining alteration (characterized by bluish-green zones distributed over the olive skin of the drupes).

Fungal biodiversity in commercial table olive packages.

In this work, a total of 72 commercial table olive packages obtained from different international markets were analysed to determine their fungal biodiversity. Viable fungal counts ranged from the detection threshold (<1.6 log10 CFU/g in 25% of cases) to a maximum of 5.86 log10 CFU/g. Assignation of fungal taxonomy was carried out through a metataxonomic analysis of the ITS region, which revealed that almost half of the total sequences obtained from all packages corresponded to the Pichia genus (44.08%), followed by Citeromyces (14.45%), Candida (8.07%), and Wickerhamomyces (6.95%). In lower proportions were also detected other genera such as Starmerella (3.60%), Saccharomyces (2.24%), Debaryomyces (2.08%), and Dekkera (2.05%). The statistical analysis allowed to link certain taxa to specific types of elaboration (lye treated, green, and black natural olives), presentation (pitted, whole, or sliced samples), and packaging material/system (glass, PET, plastic bags, and vacuum). Likewise, Zygotorulaspora genus was especially sensitive to the presence of potassium sorbate, while other genera such as Sporobolomyces, Moniliella, and Gibellulopsis were more abundant in packages treated with this preservative. Lastly, potential pathogenic fungal genera such as Alternaria, Kodamaea, Lodderomyces, Malasessia, or Aspergillus were detected in low proportions (<0.3%), although with higher representation in some individual samples. Our results contribute to improving our knowledge of the fungal population associated with this ready-to-eat fermented vegetable, providing us a strong tool to assess the safety, stability, and quality of the final product.

Killer yeasts isolated from olive brines: Technological and probiotic aptitudes.

Table olive brines, inoculated with six different starters of lactic acid bacteria (LAB) or spontaneously fermented, have been used as isolating source of killer yeasts throughout the fermentation process (120 d). Killer yeast isolates were identified and evaluated for technological and probiotic traits. Although the count of yeast population did not markedly vary among the different vessels and over time, the killer yeast phenotype was mainly present in yeast strains isolated from spontaneous fermentation; the number of killer isolates decreased over fermentation time. Killer phenotype was found in species identified as Pichia kluyveri, Zygoascus hellenicus, Wickerhamomyces anomalus, Pichia membranifaciens, Candida boidinii, Candida diddensiae and Saccharomyces cerevisiae. Among all tested isolates, W. anomalus strains evidenced the widest spectrum of enzymatic activities and the highest ß-glucosidase and phtytase activity. These strains evidenced also the best growth at low pH and increasing bile salt concentration, when grown at 37 °C, as well as the most constant viability index (%) during in vitro digestion.

From sample preparation to NMR-based metabolic profiling in food commodities: The case of table olives.

INTRODUCTION: Nuclear magnetic resonance (NMR)-based metabolic profiling has been widely used in food and plant sciences. Despite its simplicity and inherent reproducibility, the determination of the appropriate pre-processing procedures greatly affects the obtained metabolic profile. OBJECTIVES: The current study represents a detailed guide of use for untargeted NMR-based metabolic profiling of table olives (Olea europaea L.). METHODS: Greek Kalamon table olives from different geographical origins were selected as reference materials. Differently treated samples were extracted using different solvents and/or solvent systems. Chemical profiles were evaluated with high-performance thin layer chromatography (HPTLC). Different deuterated solvents and sample concentrations were evaluated for the recording of optimal quality spectra. RESULTS: The methanol extract of freeze-dried table olives was found to contain the most representative secondary metabolites, in higher concentrations, as well. The optimal deuterated solvent for the NMR analysis was methanol-d4 , while final sample concentration should be within the range of 10 to 15 mg/mL. Multivariate data analysis was also used to estimate and confirm the variation and clustering caused by different characteristics of the samples. CONCLUSIONS: Results of the present study make evident the necessity for thorough planning and method development prior to any extensive metabolomic study based on NMR spectroscopy. Pre-processing and sample preparation stages seemed to greatly affect the metabolic profile and spectral quality in the case of table olives, which by extrapolation could apply to other food commodities. Nevertheless, the nature of the samples must be fully described in general, in order to proceed to solid conclusions.

Application of Digital Olfaction for Table Olive Industry.

The International Olive Council (IOC) established that olives must be free of odors, off-flavors, and absent of abnormal ongoing alterations or fermentations. The use of electronic devices could help when classifying defects in a fast, non-destructive, cheap, and environmentally friendly way. For all of that, table olives were evaluated according to IOC regulation in order to classify the defect predominant perceiving (DPP) of the table olives and their intensity. Abnormal fermentation defects of Spanish-style table olives were assessed previously by an IOC-validated tasting panel. 'Zapateria', 'Putrid', and 'Butyric' were the defects found at different concentrations. Different volatile compounds were identified by gas chromatography in altered table olives. The same samples were measured with an electronic nose device (E-nose). E-nose data combined with chemometrics algorithms, such as PCA and PLS-DA, were able to successfully discriminate between healthy and non-healthy table olives, being this last one also separated between the first and second categories. Volatile compounds obtained with gas chromatography could be related to the E-nose measuring and sensory analysis, being capable of matching the different defects with their correspondents' volatile compounds.

Effect of High Hydrostatic Pressure in the Storage of Spanish-Style Table Olive Fermented with Olive Leaf Extract and Saccharomyces cerevisiae.

Olives treated according to the Spanish-style are firstly treated with caustic soda and then fermented in brine to reduce phenols. Next, olives are packed and subjected to pasteurization. The effect of different high hydrostatic pressure treatments (400 MPa, 4 and 6 min) was evaluated in Spanish-style table olives fermented with olive leaf extract (OLE) and S. cerevisiae compared with thermal pasteurization (P) at 80 °C for 15 min. HHP and P led to a significant reduction in yeast and aerobic mesophiles after the conservation treatment and during storage (300 days). The physical-chemical properties changed slightly during storage, except for olive hardness; olives treated with HHP presented a higher hardness than pasteurized ones. The CIELAB parameter L* decreased until day 300 in most of the treatments, as well as phenols. The HHP treatment led to significantly higher contents of phenolics (even during storage) than olives submitted to P. Some sensory attributes (colour, aspect, hardness, and overall evaluation) decreased during storage. P treatment caused a decrease in appearance, aroma, hardness, and overall evaluation compared to olives treated with HHP. Thus, the application of HHP in table olives to increase the shelf-life can be considered a valid alternative to P.

Production of volatile compounds by wild-type yeasts in a natural olive-derived culture medium.

The production of volatile compounds in naturally fermented green table olives from Manzanilla cultivar was investigated. A total of 62 volatile compounds were detected after 24 weeks of fermentation. To clarify the contribution of yeasts to the formation of these compounds, such microorganisms were isolated from the corresponding fermenting brines. Five major yeast strains were identified: Nakazawaea molendinolei NC168.1, Zygotorulaspora mrakii NC168.2, Pichia manshurica NC168.3, Candida adriatica NC168.4, and Candida boidinii NC168.5. When these yeasts were grown as pure cultures in an olive-derived culture medium, for 7 days at 25 °C, the number of volatiles produced ranged from 22 (P. manshurica NC168.3) to 60 (C. adriatica NC168.4). Contribution of each yeast strain to the qualitative volatile profile of fermenting brines ranged from 19% (P. manshurica NC168.3) to 48% (Z. mrakii NC168.2 and C. adriatica NC168.4). It was concluded that C. adriatica NC168.4 presented the best aromatic profile, being a solid candidate to be part of a novel starter culture to enhance the organoleptic properties of naturally fermented green table olives.

E-Nose Discrimination of Abnormal Fermentations in Spanish-Style Green Olives.

Current legislation in Spain indicates that table olives must be free of off-odors and off-flavors and without symptoms of ongoing alteration or abnormal fermentations. In this regard, the International Olive Council (IOC) has developed a protocol for the sensory classification of table olives according to the intensity of the predominantly perceived defect (PPD). An electronic nose (e-nose) was used to assess the abnormal fermentation defects of Spanish-style table olives that were previously classified by a tasting panel according to the IOC protocol, namely zapateria, butyric, putrid, and musty or humidity. When olives with different defects were mixed, the putrid defect had the greatest sensory impact on the others, while the butyric defect had the least sensory dominance. A total of 49 volatile compounds were identified by gas chromatography, and each defect was characterized by a specific profile. The e-nose data were analyzed using principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA). The different defects were clearly separated from each other and from the control treatment, independently of PPD intensity. Moreover, the e-nose differentiated control olives from table olives with combined sensory defects despite the dilution effect resulting from the combination. These results demonstrate that e-nose can be used as an olfactory sensor for the organoleptic classification of table olives and can successfully support the tasting panel.

Applicability of an In-Vitro Digestion Model to Assess the Bioaccessibility of Phenolic Compounds from Olive-Related Products.

The Mediterranean diet includes virgin olive oil (VOO) as the main fat and olives as snacks. In addition to providing nutritional and organoleptic properties, VOO and the fruits (olives) contain an extensive number of bioactive compounds, mainly phenolic compounds, which are considered to be powerful antioxidants. Furthermore, olive byproducts, such as olive leaves, olive pomace, and olive mill wastewater, considered also as rich sources of phenolic compounds, are now valorized due to being mainly applied in the pharmaceutical and nutraceutical industries. The digestive system must physically and chemically break down these ingested olive-related products to release their phenolic compounds, which will be further metabolized to be used by the human organism. The first purpose of this review is to provide an overview of the current status of in-vitro static digestion models for olive-related products. In this sense, the in-vitro gastrointestinal digestion methods are widely used with the following aims: (i) to study how phenolic compounds are released from their matrices and to identify structural changes of phenolic compounds after the digestion of olive fruits and oils and (ii) to support the functional value of olive leaves and byproducts generated in the olive industry by assessing their health properties before and after the gastrointestinal process. The second purpose of this review is to survey and discuss all the results available to date.

The use of multifunctional yeast-lactobacilli starter cultures improves fermentation performance of Spanish-style green table olives.

In this work, Lactobacillus pentosus LPG1, Lactobacillus pentosus Lp13, Lactobacillus plantarum Lpl15, and Wickerhanomyces anomalous Y12, all of them previously isolated from fermented table olive biofilms, were used (alone or in combination) as multifunctional starters for Manzanilla Spanish-style green table olive fermentations. Their performances were evaluated through the changes in the key physico-chemical and microbiological parameters, correlation between AI-2 production and biofilm formation, inoculum imposition, metataxonomic analysis and sensory characteristics of the finished products. Inoculation only with lactic acid bacteria (LAB) strains led to higher titratable acidities and lower pH values than the spontaneous fermentation (non-inoculated control), mainly during the first steps of processing. However, the sequential inoculation of the yeast and then the combination of the 3 LAB strains showed the most favourable evolution. LPG1 strain and, particularly Lp13, were excellent biofilms former and showed the major imposition on the fruit epidermis, as corroborated by rep-PCR analysis. Production of AI-2 was lower in the treatment inoculated exclusively with yeast Y12 but had the highest presence in the sequential yeast-LAB inoculum, with its maximum concentration and maximum LAB population on fruits (19th days) strongly related. Metataxonomic analysis of the biofilms at the end of the fermentation revealed, in addition to Lactobacillus, high proportions of sequences from genera Marinilactobacillus, Alkalibacterium, Halolactobacillus, and low levels of Halomonas and Aerococcus. Compositional data analysis of the omics data revealed that Lpl15 was scarcely efficient for controlling the spontaneous microbiota since its treatment presented the highest proportions of Aerococcus genus. Finally, the sensory analysis showed similar characteristics for the treatment inoculated with LPG1 and the spontaneous process, with olives inoculated with the yeast (alone or in combination with Lactobacillus strains) showing attractive scores. Then, inoculation of Spanish-style table olive fermentations with a sequential yeast and LAB combination could be an advisable practice.

Effect of different inoculation strategies of selected yeast and LAB cultures on Conservolea and Kalamàta table olives considering phenol content, texture, and sensory attributes.

BACKGROUND: The effects were studied of different inoculation strategies for selected starters -yeasts and lactic acid bacteria (LAB) - used for the fermentation process of two Greek olive cultivars, Conservolea and Kalamàta. The LAB strains applied were Leuconostoc mesenteroides K T5-1 and L. plantarum A 135-5; the selected yeast strains were S. cerevisiae KI 30-16 and Debaryomyces hansenii A 15-44 for Kalamàta and Conservolea olives, respectively. RESULTS: Table olive fermentation processes were monitored by performing microbiological analyses, and by monitoring changes in pH, titratable acidity and salinity, sugar consumption, and the evolution of volatile compounds. Structural modifications occurring in phenolic compounds of brine were investigated during the fermentation using liquid chromatography / diode array detection / electrospray ion trap tandem mass spectrometry (LC/DAD/ESI-MSn ) and quantified by high-performance liquid chromatography (HPLC) using a diode array detector. Phenolic compounds in processed Kalamàta olive brines consisted of phenolic acids, verbascoside, caffeoyl-6-secologanoside, comselogoside, and the dialdehydic form of decarboxymethylelenolic acid linked to hydroxytyrosol, whereas oleoside and oleoside 11-methyl ester were identified only in Conservolea olive brines. CONCLUSION: Volatile profile and sensory evaluation revealed that the 'MIX' (co-inoculum of yeast and LAB strain) inoculation strategy led to the most aromatic and acceptable Kalamàta olives. For the Conservolea table olives, the 'YL' treatment gave the most aromatic and the overall most acceptable product. © 2019 Society of Chemical Industry.

Microbial biogeography of Spanish-style green olive fermentations in the province of Seville, Spain.

The aim of this study is to examine the biogeography of the microbial communities associated to the Spanish-style green olive fermentations in the province of Seville (Andalucía, south-western Spain). Also, to understand how microorganisms colonize and persist in non-sterile food fermentations across a specific table olive producing area, i.e. a specific "agroecosystem". The microbial diversity, bacteria and yeast, in 30 ten-ton fermenters of three different fermentations yards (patios) along the olive fermentation was studied. A total of 951 microbial isolates were obtained which were clustered according to their RAPD profile. A total of 376 distinct genotypes were identified, belonging to 57 different microbial species, 41 bacterial and 16 yeast species. Up to 16 bacterial species had not been described before in table olives. Only the species Lactobacillus pentosus showed a ubiquitous presence in all 30 fermenters. Pediococcus parvulus, Lactobacillus collinoides/paracollinoides, Lactobacillus coryniformis, Lactobacillus plantarum, Pichia manshurica and Candida thaimueangensis were found in every patio. Cosmopolitan strains, up to 15, were shared by the three patios and belonged to the species L. pentosus (12 strains), P. parvulus (1), L. collinoides/paracollinoides (1) and P. manshurica (1). To expand our biodiversity analyses to the "regional" level, we have compared our results with those obtained from two previously studied patios of similar characteristics and in the same geographical area. PERMANOVA analysis of the microbial community composition revealed significant differences among different patios in their structure at every fermentation stage. In contrast, SIMPER analyses showed that, as fermentation progressed, the overall dissimilarities among patios were reduced. Discriminant species were identified for each fermentation stage. Among these, L. pentosus and P. parvulus were "eu-constant" species, while L. collinides/paracollinoides and Marinilactibacillus psychrotolerans group were "constant" species that could be considered microbial key taxa based on the occurrence stability index. The characteristic and, presumably, well adapted microbiota associated to the Spanish-style olive fermentations at the specific geographic area described here is a valuable natural resource which should be preserved conveniently. To our knowledge, this is the first study on the microbial biogeography of table olive fermentations, both at the species and strain levels.