Functional features of the exopolysaccharide extracts produced by Lactiplantibacillus strains isolated from table olives.

This study evaluates the functional characteristics of the exopolysaccharide (EPS) extracts produced by various strains of Lactiplantibacillus pentosus (LPG1, 119, 13B4, and Lp13) and Lactiplantibacillus plantarum (Lp15) isolated from table olives. None of the EPS crude extracts showed cytotoxicity when administered to THP-1 human macrophage cells at dosages ranging from 6.25 to 50 µg mL-1. Many exhibited anti-inflammatory properties (reduction of pro-inflammatory cytokines TNF-α and IL-6 production) and antioxidant activity (reduction of ROS%) when macrophages were stimulated with Escherichia coli lipopolysaccharide. Notably, the EPS extract produced by the L. pentosus LPG1 strain had the best results corroborated by western blot immune analysis for differential expression of COX-2, Nrf-2, and HO-1 proteins, with the most significant antioxidant and anti-inflammatory response observed at a dosage of 50 µg mL-1. Chemical analysis revealed that the EPS extract produced by this strain contains a heteropolymer composed of mannose (35.45%), glucose (32.99%), arabinose (17.93%), xylose (7.48%), galactose (4.03%), rhamnose (1.34%), and fucose (0.77%). Finally, we conducted response surface methodology to model the EPS extract production by L. pentosus LPG1 considering pH (3.48-8.52), temperature (16.59-33.41 °C) and salt concentration (0.03-8.77% NaCl) as independent variables. The model identified linear effects of salt and pH and quadratic effects of salt as significant terms. The maximum EPS extract production (566 mg L-1) in a synthetic culture medium (MRS) was achieved at pH 7.5, salt 7.0%, and a temperature of 20 °C. These findings suggest the potential for novel applications for the EPS produced by L. pentosus LPG1 as nutraceutical candidates for use in human diets.

Impact of Salts Mixtures on the Physicochemical and Sensory Characteristics of Spanish-Style Manzanilla Green Table Olives during Packaging.

Using response surface methodology (RSM), this study investigates the effect of NaCl substitution (50%) with KCl, CaCl2, and MgCl2 in the packaging brines (controlled variables) on the characteristics (responses) of plain green Spanish-style Manzanilla olives, maintaining the salt-mixture level of 5%. The RSM showed that the increment of CaCl2 caused a linear significant (p-value ≤ 0.05) decrease in pH and a linear increase in firmness (instrumental), hardness (panel scores), and crunchiness. The models for bitterness and fibrousness also included quadratic (CaCl2·MgCl2) and cubic (the three salt) interactions, which led to areas of minimum and maximum scores around the central points of the CaCl2-MgCl2 and KCl-MgCl2 axes, respectively. In contrast, the increase in the KCl level linearly decreased bitterness scores. Optimisation resulted in a relatively low desirability (0.57) and the selection of a combination that may necessitate further refinement, such as increasing KCl or reducing CaCl2 levels, especially for markets sensitive to bitterness. Interestingly, the overall score and buying predisposition positively correlate with salty, smell, acid, and appearance and negatively with bitterness. Furthermore, PLS-R analysis found that the pivotal attributes influencing overall appreciation were smell and crunchiness while buying predisposition was promoted by crunchiness. Conversely, bitterness had a detrimental impact on these appreciations. Cluster analysis grouped the experimental runs into four categories, with sensory profiles predominantly diverging in bitterness, salty, and kinesthetic characteristics. Ultimately, this study elucidates four distinct sensory profiles that consumers experience.

Application of Compositional Data Analysis to Study the Relationship between Bacterial Diversity in Human Faeces and Sex, Age, and Weight.

This work uses Compositional Data Analysis (CoDA) to examine the typical human faecal bacterial diversity in 39 healthy volunteers from the Andalusian region (Spain). Stool samples were subjected to high-throughput sequencing of the V3 and V4 regions of the 16S ribosomal RNA gene using Illumina MiSeq. The numbers of sequences per sample and their genus-level assignment were carried out using the Phyloseq R package. The alpha diversity indices of the faecal bacterial population were not influenced by the volunteer's sex (male or female), age (19-46 years), and weight (48.6-99.0 kg). To study the relationship between these variables and the faecal bacterial population, the ALDEx2 and coda4microbiome CoDA packages were used. Applying ALDEx2, a trend suggesting a connection between sex and the genera Senegalimassilia and Negatibacillus (slightly more abundant in females) and Desulfovibrio (more abundant in males) was found. Moreover, age was tentatively associated with Streptococcus, Tizzerella, and Ruminococaceae_UCG-003, while weight was linked to Senegalimassilia. The exploratory tool of the coda4microbiome package revealed numerous bacterial log-ratios strongly related to sex and, to a lesser extent, age and weight. Moreover, the cross-sectional analysis identified bacterial signature balances able to assign sex to samples regardless of controlling for volunteers' age or weight. Desulfovibrio, Faecalitalea, and Romboutsia were relevant in the numerator, while Coprococcus, Streptococcus, and Negatibacillus were prominent in the denominator; the greater presence of these could characterise the female sex. Predictions for age included Caproiciproducens, Coprobacter, and Ruminoclostridium in the numerator and Odoribacter, Ezakiella, and Tyzzerella in the denominator. The predictions depend on the relationship between both groups, but the abundance of the first group and scarcity of the second could be related to older individuals. However, the association of the faecal bacterial population with weight did not yield a satisfactory model, indicating scarce influence. These results demonstrate the usefulness of the CoDA methodology for studying metagenomics data and, specifically, human microbiota.

Modeling the antimicrobial effects of olive mill waste extract, rich in hydroxytyrosol, on the growth of lactic acid bacteria using response surface methodology.

The objective of this study is to assess the inhibitory effects of an aqueous extract from olive oil mill waste (alperujo) on the growth of a lactic acid bacteria (LAB) cocktail consisting of various strains of Lactiplantibacillus pentosus and Lactiplantibacillus plantarum species. For this purpose, response surface methodology was employed using two independent variables (pH levels 3.5-5.55; hydroxytyrosol concentration ranging from 0.93-2990 ppm). The response variable was the average inhibition per treatment on the LAB cocktail (expressed as a percentage). The developed model identified significant terms, including the linear effect of hydroxytyrosol and pH, their interaction, and the quadratic effect of pH. Maximum inhibition of the LAB cocktail was observed at progressively higher concentrations of hydroxytyrosol and lower pH values. Therefore, complete inhibition of LAB in the synthetic culture medium could only be achieved for concentrations of 2984 ppm hydroxytyrosol at a pH of 3.95. These findings suggest that extracts derived from "alperujo" could be utilized as a natural preservative in acidified foods with a bitter flavor and antioxidant requirements.

The Desalting Process for Table Olives and Its Effect on Their Physicochemical Characteristics and Nutrient Mineral Content.

The desalting process is critical for packaging table olives in brine with reduced NaCl or fortified mineral nutrients. In this study, the effect of desalting on the physicochemical characteristics and mineral content of green Manzanilla Spanish-style (plain and stuffed with pepper paste) and DOP Aloreña de Málaga table olives was investigated for the first time. The surface colour of the fruits turned slightly brownish, and the olives became somewhat softer. The lactic acid, the mineral macronutrients (mainly) and micronutrient contents decreased, while flesh moisture increased. The kinetic parameters of the minerals' losses depended on the presentation, with the estimated values for plain olives being the lowest (slowest desalting). Overall, the desalting process resulted in slight quality damage and a moderated decrease in the mineral concentration in the flesh, leading to some product degradation. This study provides quantitative information on these changes that may affect the commercial value of the final products and offers information for viable designs.

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.

Oral intake of Lactiplantibacillus pentosus LPG1 Produces a Beneficial Regulation of Gut Microbiota in Healthy Persons: A Randomised, Placebo-Controlled, Single-Blind Trial.

The search for vegetable-origin probiotic microorganisms is a recent area of interest. This study conducted a phase I clinical trial to assess the effects of oral administration of Lactiplantibacillus pentosus LPG1, a natural strain with probiotic potential isolated from table olive fermentations, on the gut microbiota. The trial was a randomised, placebo-controlled, single-blind study involving 39 healthy volunteers. Group A (n = 20) ingested one capsule/day of L. pentosus LPG1 containing 1 × 1010 UFC/capsule, while Group B (n = 19) received one capsule/day containing only dextrose (placebo). The capsules were taken during breakfast for 30 consecutive days. Human stool samples were collected from all volunteers at the beginning (baseline) and at the end of the study (post-intervention) and were subjected to 16S rRNA metataxonomic analysis using Illumina MiSeq. Sequencing data at the genus level were statistically analysed using traditional methods and compositional data analysis (CoDA). After treatment, the alpha diversity in Group B (placebo) decreased according to an increase in the Berger and Parker dominance index (p-value < 0.05); moreover, dominance D increased and Simpson 1-D index decreased (p-value < 0.10). The Lactobacillus genus in the faeces was included in the CoDA signature balances (selbal and coda4microbiome) and played a notable role in distinguishing samples from baseline and post-intervention in Group A (LPG1). Additionally, ingesting L. pentosus LPG1 modified the gut microbiota post-intervention, increasing the presence of Parabacteroides and Agathobacter, but reducing Prevotella. These findings suggest that L. pentosus LPG1 is a potentially beneficial gut microbiota modulator in healthy persons.

Effect of Spanish-Style Table Olive Processing on Fatty Acid Profile: A Compositional Data Analysis (CoDA) Approach.

This manuscript considers that the composition of Manzanilla and Hojiblanca fats are compositional data (CoDa). Thus, the work applies CoDa analysis (CoDA) to investigate the effect of processing and packaging on the fatty acid profiles of these cultivars. To this aim, the values of the fat components in percentages were successively subjected to exploratory CoDA tools and, later, transformed into ilr (isometric log-ratio) coordinates in the Euclidean space, where they were subjected to the standard multivariate techniques. The results from the first approach (bar plots of geometric means, tetrahedral plots, compositional biplots, and balance dendrograms) showed that the effect of processing was limited while most of the variability among the fatty acid (FA) profiles was due to cultivars. The application of the standard multivariate methods (i.e., Canonical variates, Linear Discriminant Analysis (LDA), ANOVA/MANOVA with bootstrapping and n = 1000, and nested General Linear Model (GLM)) to the ilr coordinates transformed data, following Ward's clustering or descending order of variances criteria, showed similar effects to the exploratory analysis but also showed that Hojiblanca was more sensitive to fat modifications than Manzanilla. On the contrary, associating GLM changes in ilr with fatty acids was not straightforward because of the complex deduction of some coordinates. Therefore, according to the CoDA, table olive fatty acid profiles are scarcely affected by Spanish-style processing compared with the differences between cultivars. This work has demonstrated that CoDA could be successfully applied to study the fatty acid profiles of olive fat and olive oils and may represent a model for the statistical analysis of other fats, with the advantage of applying appropriate statistical techniques and preventing misinterpretations.

Quality parameters and fatty acid and triacylglycerol profiles of directly brined table olive fat throughout processing and shelf life.

The transformations that may suffer directly brined table olive fat during processing were studied using cracked Aloreña de Málaga olive as a model. The classical studies showed that storage increased acidity and K270, but not peroxide value, K232 and ΔK. FA profiles, nutritional fat subclasses, and TAGs suffered several significant changes along processing, although some could be spurious. Compositional data (CoDa) analysis identified C18:2n-6 and C18:1c, and their corresponding TAGs as the most affected compounds, conclusion that was particularly evident after weighted CoDa log-ratio analysis (LRA). Thus, CoDa analysis is a promising alternative statistical tool to study table olive FA, TAG profiles and fats in general. Most of the quality parameters' values and FA and TAG concentrations were compatible with Virgin Olive Oil (VOO), showing that the fat quality and nutritional characteristics of natural green olives in brine could be comparable to that of VOO.

Delving into the bacterial diversity of spoiled green Manzanilla Spanish-style table olive fermentations.

This work applies metataxonomic, standard statistics, and compositional data (CoDa) techniques to study the bacterial diversity of spoiled and normal Spanish-style table olive fermentations, analysing a total of 10-tons of industrial fermentation containers from two processing yards. Forty percent were affected by butyric, sulfidic, or putrid spoilage, while 60% followed the ordinary fermentation course. The samples were obtained at 30 days of fermentation, determining their 16S rRNA gene Amplicon Sequence Variant compositions (ASVs). The butyric containers showed a bacterial profile strongly associated with the genera Enterococcus, Leuconostoc, and Atlantibacter, but also with Lactiplantibacillus and Melissococcus, and less confident to Raoultella, Enterobacter, Serratia, and Celerinatantimonas. The sulfidic fermentation was linked to Alkalibacterium and, to a lesser extent, Marinilactibacillus and the absence of Lactiplantibacillus. Putrid spoilage was mainly related to Halolactibacillus and Alkalibacterium. Sulfidic/putrid (together) differed from butyric spoilage by the presence of Alkalibacterium/Marinilactibacillus as well as by Halomonas/Halanaerobium. Lactiplantibacillus dominated normal fermentations, but Vibrio was also frequently found (0-46%), apparently not causing any alteration. These results contribute to a better microbial characterisation of non-zapatera spoiled table olive fermentations. They also suggest using several statistical techniques to discriminate normal vs spoiled fermentations adequately.

Multi-Statistical Approach for the Study of Volatile Compounds of Industrial Spoiled Manzanilla Spanish-Style Table Olive Fermentations.

Table olives can suffer different types of spoilage during fermentation. In this work, a multi-statistical approach (standard and compositional data analysis) was used for the study of the volatile organic compounds (VOCs) associated with altered (butyric, sulfidic, and putrid) and non-altered (normal) Manzanilla Spanish-style table olive fermentations. Samples were collected from two industrial fermentation yards in Seville (Spain) in the 2019/2020 season. The VOC profiles of altered (n = 4) and non-altered (n = 6) samples were obtained by headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Ninety-one VOCs were identified and grouped into alcohols (30), esters (21), carbonyl compounds (12), acids (10), terpenes (6), phenols (6), sulfur compounds (2), and others (4). The association of the VOCs with spoilage samples depended on the standard or compositional statistical methodology used. However, butyric spoilage was strongly linked by several techniques to methyl butanoate, ethyl butanoate, and butanoic acid; sulfidic spoilage with 2-propyl-1-pentanol, dimethyl sulfide, methanol, 2-methylbutanal, 2-methyl-2-butenal, ethanol, 2-methyl-3-buten-2-ol, and isopentanol, while putrid was mainly related to D-limonene and 2-pentanol. Our data contribute to a better characterisation of non-zapatera spoiled table olive fermentations and show the convenience of using diverse statistical techniques for a most robust selection of spoilage VOC markers.

Effect of season, feeding, and anatomical region on the triacylglycerol profile of Iberian pig fat.

The work studies the effects of season, feeding type, and anatomical region on the Iberian pig fat triacylglycerol (TAG) profiles, considered as compositional data (CoDa). The analysis consisted of applying exploratory tools in the simplex and standard multivariate techniques to data transformed into the Euclidean space (ilr coordinates). Compositional biplot showed differences in TAG containing palmitic (P) and oleic (O) acids between the 2005 and 2003/2004 seasons but not within these. PPP (clr variance, 0.139), OLL (0.095), PPS (0.075), POPo + PLP (0.074), and PSS (0.629) showed high CoDa variability among treatments. The ANOVA analysis found significant (p ≤ 0.05) effects of season, feeding type, and anatomical region on pig fat TAG profile, but only that for 2005 season was well predicted (97.5%) by discriminant analysis (DA). Overall, season was more influential on the Iberian pig fat TAG profile than montanera length and sampling region, which effects were not significant for some statistical techniques.

Relating starter cultures to volatile profile and potential markers in green Spanish-style table olives by compositional data analysis.

This work relates native lactic acid bacteria (LAB) (Lactobacillus pentosus LPG1, L. pentosus Lp13, and Lactobacillus plantarum Lpl15) and yeast (Wickerhamomyces anomalus Y12) starters to the volatile components (VOCs) produced in green Spanish-style table olives. For this aim, the VOC profile was considered as compositional data (CoDa). The CoDa analysis generated new information on the relationship among inocula and VOCs through the tetrahedral plot, CoDa-biplot, variation array matrix, and CoDa dendrogram. The ilr (which includes pivot) coordinates (Euclidean space) from VOCs produced more reliable starters' clustering than the original data. The potential VOC markers, identified by a test based on the pairwise comparison of the logratio variation arrays from the whole data set and the individual groups, were (starters in the parenthesis): 2-phenylethyl acetate (LPG1, Y12, Y12 + LAB), methanol (Lpl15), cis-2-penten-1-ol (LPG1, Y12, Y12 + LAB), 2-methyl-3-hexanol (LPG1, Y12), U (non-identified) C (m/z 83-112-97) (Y12) and UF (m/z 95-154-110) (LPG1, Y12 + LAB). Besides, some VOCs were partial/totally inhibited by specific starters: 2-methyl-1-propanol (Lp13, Y12 + LAB), 2-phenyl ethanol (Lp13), furfuryl methyl ether (Y12 + LAB), purpurocatechol (Y12, Y12 + LAB), 4-ethyl guaiacol (Lp13, Lpl15), 4-ethyl phenol (Lpl15), 5-tert-butylpyrogallol (Lp13, Lpl15), and UE (m/z 111-198) (Lp13). A better understanding of the relationship between starters and their VOC may facilitate modelling the flavour and quality of Spanish-style green table olive fermentations.

Exploring bacteria diversity in commercialized table olive biofilms by metataxonomic and compositional data analysis.

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%), and Pediococcus (26.09%). Celerinatantimonas, Leuconostoc, Alkalibacterium, Pseudomonas, Marinilactibacillus, Weissella, and the family Enterobacteriaceae were also present in at least 80% of samples. Regarding foodborne pathogens, only Enterobacteriaceae, Vibrio, and Staphylococcus were 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, and Alkalibacterium were 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.

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.

In Vitro Bioaccessibility of Ripe Table Olive Mineral Nutrients.

For the first time, the bioaccessibility of the mineral nutrients in ripe table olives and their contributions to the recommended daily intake (RDI), according to digestion methods (Miller's vs. Crews' protocols), digestion type (standard vs. modified, standard plus a post-digest re-extraction), and mineralisation system (wet vs. ashing) were studied. Overall, when the standard application was used, Miller's protocol resulted in higher bioaccessibilities of Na, K, Ca, Mg, and Fe than the Crews' method. The modified protocols improved most of these values, but the Crews' results only approximated the Miller's levels in the case of Na and K. The bioaccessibility of P was hardly affected by the factors studied, except that the modified Miller's protocol led to higher levels when ashing. No significant effect of the mineralisation system was found. The modified Miller's protocol, regardless of the mineralisation system, led to the overall highest bioaccessibility values in ripe olives, which were: Na (96%), K (95%), Ca (20%), Mg (73%), Fe (45%), and P (60%). Their potential contributions to the RDI, based on these bioaccessibilities and 100 g olive flesh service size, were then 29, 0.5, 4, 3, 33, and 1% respectively. The investigation has led to the development of a method for assessing the bioaccessibility of the mineral nutrients not only in ripe but also in the remaining table olive presentations and opens a new research line of great interest for producing healthier products.

Influence of Acid Adaptation on the Probability of Germination of Clostridium sporogenes Spores Against pH, NaCl and Time.

The Clostridium sp. is a large group of spore-forming, facultative or strictly anaerobic, Gram-positive bacteria that can produce food poisoning. The table olive industry is demanding alternative formulations to respond to market demand for the reduction of acidity and salt contents in final products. while maintaining the appearance of freshness of fruits. In this work, logistic regression models for non-adapted and acid-adapted Clostridium sp. strains were developed in laboratory medium to study the influence of pH, NaCl (%) and time on the probability of germination of their spores. A Clostridium sporogenes cocktail was not able to germinate at pH < 5.0, although the adaptation of the strains produced an increase in the probability of germination at 5.0-5.5 pH levels and 6% NaCl concentration. At acidic pH values (5.0), the adapted strains germinated after 10 days of incubation, while those which were non-adapted required 15 days. At pH 5.75 and with 4% NaCl, germination of the adapted strains took place before 7 days, while several replicates of the non-adapted strains did not germinate after 42 days of storage. The model was validated in natural green olive brines with good results (>81.7% correct prediction cases). The information will be useful for the industry and administration to assess the safety risk in the formulation of new processing conditions in table olives and other fermented vegetables.

Impact of Processing and Extraction on the Minor Components of Green Spanish-Style Gordal Table Olive Fat, as Assessed by Innovative Approaches.

This work aims to study the effect of the green Spanish-style table olive processing and extraction method of fat on its minor components. For this purpose, it uses standard multivariate analysis (developed for Euclidean space), Compositional Data (CoDa) analysis (for data in the simplex) and Multiple Factor analysis (MFA). Overall, processing had a scarce effect on most of the minor components except ethyl and methyl esters and diacylglycerols, which markedly increased during fermentation; however, these compounds in table olive do not have the negative connotations that those in olive oil do since they are normal metabolites from the yeast microflora habitually present during the process. The work also showed that the quantification of minor components in table olive fat was an extraction-dependent method since Soxhlet increased the concentrations of fatty alcohols, triterpene dialcohols, sterols, waxes and polar compounds. Regarding statistical methods, CoDa analysis strategies were successfully applied to produce more appropriate clustering and Principal Component Analysis (PCA) segregation than standard tools. Moreover, MFA allowed for study of the components individually and by groups; the relationships among groups led to the most appropriate clustering and PCA segregation of samples and revealed the effect of the chemical groups' evolution on the similarity/dissimilarity between samples. Therefore, MFA was the statistical analysis that led to the most information on the effect of processing and extraction methods. Its combination with appropriate CoDa logratios could be an exciting challenge.

Panel and Panelist Performance in the Sensory Evaluation of Black Ripe Olives from Spanish Manzanilla and Hojiblanca Cultivars.

There is vast experience in the application of sensory analysis to green Spanish-style olives, but ripe black olives (≈1 × 106 kg for 2016/2017) have received scarce attention and panelists have less experience on the evaluation of this presentation. Therefore, the study of their performance during the assessment of this presentation is critical. Using previously developed lexicon, ripe olives from Manzanilla and Hojiblanca cultivars from different origins were sensory analysed according to the Quantitative Descriptive Analysis (QDA). The panel (eight men and six women) was trained, and the QDA tests were performed following similar recommendations than for green olives. The data were examined while using SensoMineR v.1.07, programmed in R, which provides a diversity of easy to interpret graphical outputs. The repeatability and reproducibility of panel and panelists were good for product characterisation. However, the panel performance investigation was essential in detecting details of panel work (detection of panelists with low discriminant power, those that have interpreted the scale in a different way than the whole panel, the identification of panelists who required training in several/specific descriptors, or those with low discriminant power). Besides, the study identified the descriptors of hard evaluation (skin green, vinegar, bitterness, or natural fruity/floral).

Lactic Acid Bacteria and Yeast Inocula Modulate the Volatile Profile of Spanish-Style Green Table 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.