Table Olive Fermentation Research Articles

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.

Assessing Listeria monocytogenes growth during Spanish-style green table olive fermentation

Table olives are one of the most important fermented vegetables in the Mediterranean countries. Although this fermented vegetable has a long record of microbial safety, various studies have shown the presence of Listeria monocytogenes during table olive processing. This study focused on evaluating the behaviour of a cocktail of L. monocytogenes strains during fermentation of Spanish-style green Manzanilla olives. For this purpose, a challenge test consisting of artificial inoculation of olive fermentation with high levels of the food-borne pathogen (7.0 log10 CFU/ml) was performed. The Fermentation was carried out at the laboratory scale in sealed glass bottles (n = 168) under controlled conditions to avoid the initiation of lactic acid fermentation intentionally. This point was confirmed by the evolution of pH, which ranged from an initial 9.0 to final 6.0 units (decrease kinetic constant of −0.0016 h−1), as well as by the behaviour of lactic acid and mesophilic bacteria during the study (55 days). The fate of L. monocytogenes during fermentation was described using mathematical models, including the two-term Gompertz model. During this time, the food-borne pathogen showed an initial growth phase which lasted until 250 h, reaching a maximum population level close to 9.0 log10 CFU/ml, even in the presence of high salt concentrations (>6.0% NaCl). Then, a rapid decline phase was observed but continuously detecting a residual tail of around 2 log10 CFU/ml. Data obtained in this work shows the importance of the control of olive fermentation, especially during the first weeks when pH is alkaline-neutral and environmental conditions could favour L. monocytogenes growth.

Fermented table olives from Cyprus: Microbiota profile of three varieties from different regions through metabarcoding sequencing.

The knowledge about the microbial diversity of different olives varieties from diverse regions in the Mediterranean basin is limited. This work aimed to determine the microbial diversity of three different fermented olive varieties, collected from different regions in Cyprus, via Next Generation Sequencing (NGS) analysis. Olives were spontaneously fermented for 120 days, microbial DNA was extracted from the final products, and subjected to 16S rRNA gene and ITS1 loci metabarcoding analysis for the determination of bacterial and fungal communities, respectively. Results revealed that the bacterial profile of the studied varieties was similar, while no noteworthy differences were observed in olives from different regions. The bacterial profile was dominated by the co-existence of Lactobacillus and Streptococcus, while the genera Lactococcus and Salinivibrio and the family Leuconostocaceae were also present in increased relative abundances. Regarding fungal communities, the analysis indicated discrimination among the different varieties, especially in Kalamata ones. The most abundant fungi were mainly the genera Aspergillus, Botryosphaeria, Meyerozyma, and Zygosaccharomyces for Cypriot olives, the genera Botryosphaeria, Saccharomyces, Geosmithia, and Wickeromyces for Kalamata variety, while the dominant fungi in the Picual variety were mainly members of the genera Candida, Penicillium, Saccharomyces, Hanseniospora and Botryosphaeria. Potential microbial biomarkers that distinguish the three varieties are also proposed. Moreover, interaction networks analysis identified interactions among the key taxa of the communities. Overall, the present work provides useful information and sheds light on an understudied field, such as the comparison of microbiota profiles of different varieties from several regions in Cyprus. The study enriches our knowledge and highlights the similarities and the main differences between those aspects, booming in parallel the need for further works on this frontier, in the attempt to determine potentially olives' microbial terroir in Cyprus. Our work should be used as a benchmark for future works in this direction.

Cobrançosa Table Olive Fermentation as per the Portuguese Traditional Method, Using Potentially Probiotic Lactiplantibacillus pentosus i106 upon Alternative Inoculation Strategies

Spontaneous fermentation of table olives, as per a traditional Mediterranean process, is still performed empirically; hence, final product quality is somewhat unpredictable. Our main goal was to validate an endogenous (potentially probiotic) lactic acid bacterium strain in Cobrançosa table olives as a vector for a more standardized process, further adding commercial value to the olives themselves. The traditional Portuguese fermentation process typically consists of two stages: sweetening, when olives are periodically washed with spring water to different proportions, and salting, when water is no longer changed, but salt is gradually added to the brine, up to 7–10% (w/w). Lactiplantibacillus pentosus i106 was inoculated as follows: (plan A) 2020/21 harvest, with 0, 3, 5, and 7% (w/v) NaCl, without sweetening; (plan B) 2020/21 harvest, with 5 and 7% (w/v) NaCl, during salting and sweetening; and (plan C) 2019/20 harvest, with 5% (w/v) salt, and sweetening and salting. Microbiological, physical, and biochemical evolutions were monitored for 8 months, and final nutritional and sensory features were duly assessed. Compared to the control, lactic acid bacteria (LAB) predominated over yeasts only if deliberately inoculated; however strain viability was hindered above 5% (w/w) NaCl, and LAB inhibited enterobacteria. Degradation of (bitter) oleuropein to hydroxytyrosol and verbascoside was faster upon inoculation. Color-changing olives from the 2020/21 harvest exhibited higher fat content and lower water content compared to green ones (2019/20 harvest), and different salt levels and inoculation moments produced distinct sensory properties. The best protocol was plan C, in terms of overall eating quality; hence, the addition of Lpb. pentosus i106 provides benefits as a supplementary additive (or adjunct culture), rather than a starter culture.

Fermentation of cv. Kalamata Natural Black Olives with Potential Multifunctional Yeast Starters.

The purpose of this study was to explore the inoculated fermentation of cv. Kalamata natural black olives using selected strains of yeast cultures with multifunctional potential. For this purpose, five yeast starters belonging to Candida boidinii (four starters) and Saccharomyces cerevisiae (one starter), previously isolated from table olive fermentation of the same variety and screened for their technological characteristics and probiotic potential, were inoculated in brines at the beginning of fermentation. Microbial populations (lactic acid bacteria, yeasts, and Enterobacteriaceae), pH, titratable acidity, organic acids, and ethanol were monitored during fermentation for a period of 5 months. At the same time, the survival of each starter was assessed by culture-dependent molecular identification at the beginning (0 days), middle (75 days), and final stages (150 days) of fermentation in the brines and olives (at the end of the process only). The results revealed the coexistence of yeasts and lactic acid bacteria (LAB) throughout fermentation in most processes and also the absence of Enterobacteriaceae after the first 20 days of brining. The population of yeasts remained 2 log cycles below LAB counts, except for in the inoculated treatment with C. boidinii Y28, where the yeast starter prevailed from day 60 until the end of the fermentation, as well as in the inoculated treatment with C. boidinii Y30, where no LAB could be detected in the brines after 38 days. At the end of the process, LAB ranged between 4.6 and 6.8 log10 CFU/mL, while yeasts were close to 5.0 log10 CFU/mL, except for the inoculated fermentation with C. boidinii Y27 and spontaneous fermentation (control), in which the yeast counts were close to 3.5 log10 CFU/mL. At the end of fermentation, the recovery percentage of C. boidinii Y27 was 50% in the brines and 45% in the olives. C. boidinii Y28 and S. cerevisiae Y34 could be recovered at 25% and 5% in the brine, respectively, whereas neither starter could be detected in the olives. For C. boidinii Y30, the recovery percentage was 25% in the brine and 10% in the olives. Finally, C. boidinii Y31 could not be detected in the brines and survived at a low percentage (10%) in the olives.

Influence of 1-Methylcyclopropene (1-MCP) on the Processing and Microbial Communities of Spanish-Style and Directly Brined Green Table Olive Fermentations

This work evaluates the effect of 1-methylcyclopropene (1-MCP) on postharvest and fermentation of Manzanilla cultivar, processed as Spanish-style or directly brined table olives. During postharvest handling, 1-MCP (2.85 µL/L) reduced the number of colour-turning olives by 18.42% over the untreated fruits. In Spanish-style and directly brined fermentation, the 1-MCP treatment led to lower pH levels, higher titratable acidities, improved firmness and colour olives than untreated fruits. A panel of expert testers also gave higher scores, and overall acceptability to the 1-MCP treated fruits, especially in the case of Spanish-style fermented olives. Metagenomic analysis of olive biofilms at the end of the fermentation process (176 days) revealed that Lactiplantibacillus was the most abundant bacterial genus in both Spanish-style and directly brined olives (>72%). However, fungal biodiversity was higher than bacterial in all treatments. Saccharomyces was the predominant yeast genus associated with directly brined olives (>97%), whilst Wickerhamomyces (>37%) and Zygoascus (>18%) were with Spanish-style fermentations. The 1-MCP treatment doubled the presence of Wickerhamomyces in Spanish-style fruits (74%) whilst reducing the presence of Zygoascus and allowing the growth of Enterobacter (15%) in directly brined olives. Thus, the postharvesting treatment of table olives with 1-MCP could help reduce the maturation progress of olives and improve the organoleptic and quality characteristics of the products without affecting the microbiological evolution of the fermentations.

Application of Electronic Nose to Discriminate Species of Mold Strains in Synthetic Brines.

The chemical composition of the brine for Spanish-style table olives plays a crucial role during the fermentation process. Traditional laboratory analysis requires a high consumption of reagents, highly qualified personnel, sophisticated equipment, long analysis times, and large amounts of samples. Analysis carried out using an electronic nose (E-nose) offers an alternative, non-destructive technique and is useful in determining alterations in brines caused by microorganisms. In the present research, nine mold strains isolated from spoiled olives were inoculated in synthetic brines to determine the effect of microbial development on sensory quality, volatile profile, and the capacity of E-nose to discriminate altered brines from the healthy ones. The brines inoculated with the mold strains presented negative attributes related to aromas of mold, wood, leather, rancidity and, organic solvents among others. The highest intensity of defect was presented by the brines inoculated with the strains Galactomyces geotricum (G.G.2); three Penicillium expansum (P.E.3, P.E.4, and P.E.20); one Penicillium glabrum (P.G.19); three Aspergillus flavus (A.F.9, A.F.18, and A.F.21); and one Fusarium solani (F.S.11). A total of 19 volatile compounds were identified by gas chromatography. Sensory analysis allowed us to classify the synthetic brines based on the degree of alteration produced by the mold strains used. Also, the E-nose data were able to discriminate the inoculated brines regardless of the intensity of the defect. These results demonstrate the capacity of the E-nose to discriminate alterations in brines produced by molds, thereby making it a useful tool to be applied during the elaboration process to detect early alterations in table olive fermentation.

Bacterial metataxonomic analysis of industrial Spanish-style green table olive fermentations

In this work, metataxonomic-compositional data analysis was applied to study the influence of inoculation, type of ecosystem (brine or fruit), and time of fermentation (0, 24, and 83 days) on the bacterial composition dynamic during industrial Manzanilla Spanish-style green table olive fermentations. Differences in the physicochemical and microbiological characteristics between inoculated and spontaneous fermentations were scarce, except for the high lactic acid bacteria population (>10 million CFU/g) in the inoculated olive biofilms. The metataxonomic analysis showed eight relevant taxa (Lactiplantibacillus, Vibrio, Halolactibacillus, Aerococcus, Catenococcus, Salinivibrio, Alkalibacterium, and Marinilactibacillus) and absence of pathogens, but the microbiota composition markedly changed throughout fermentation. At the process onset, the main genera were Vibrio, Halolactibacillus, Marinilactibacillus, and Alkalibacterium, while Lactiplantibacillus was almost absent. Regardless of inoculation, Lactiplantibacillus became the predominant genus as the fermentation progressed, albeit Vibrio, Marinilactibacillus, Alkalibacterium, and Halolactibacillus also persisted until the end of the process (but did not cause spoilage). Ferrimonas genus was detected for the first time in table olives, being found in 71% of the samples. Besides, there was more bacterial biodiversity in the brine ecosystem than olive biofilms. Overall, inoculation led to more homogenous and equilibrated bacterial populations at the end of the process. This study expands the knowledge on the bacterial population changes occurring during industrial table olive fermentation, the differences between populations in brines and biofilms, and the beneficial effects of inoculation for using table olives as a carrier of beneficial microorganisms.

Selection of Lactiplantibacillus Strains for the Production of Fermented Table Olives.

Lactiplantibacillus strains (n. 77) were screened for technological properties (e.g., xylose fermentation, EPS production, antimicrobial activity, tolerance to NaCl and phenolic compounds, oleuropein degradation and hydroxytyrosol formation) relevant for the production of fermented table olives. Survival to olive mill wastewater (OMW) and to simulated gastro-intestinal tract (GIT), the capability to grow at different combinations of NaCl and pH values, radical scavenging activities and biofilm formation were further investigated in 15 selected strains. The screening step revealed high diversity among Lactiplantibacillus strains. Most of the strains were able to ferment xylose, while only a few strains produced EPS and had inhibitory activity against Y. lipolytica. Resistance to phenolic compounds (gallic, protocatechuic, hydroxybenzoic and syringic acids), as well as the ability to release hydroxytyrosol from oleuropein, was strain-specific. OMWs impaired the survival of selected strains, while combinations of NaCl ≤ 6% and pH ≥ 4.0 were well tolerated. DPPH and hydroxyl radical degradation were strain-dependent, while the capability to form biofilm was affected by incubation time. Strains were very tolerant to the GIT. The genome of Lpb. pentosus O17 was sequenced and analysed to verify the presence of genes involved in the degradation and metabolism of phenolic compounds. O17 lacks carboxylesterase and gallate decarboxylase (subunits B and D) sequences, and its gene profile differs from that of other publicly available Lpb. pentosus genomes.

Brine salt concentration reduction and inoculation with autochthonous consortia: Impact on Protected Designation of Origin Nyons black table olive fermentations

Nyons table olives, named after the French city where they are processed, are naturally fermented black table olives. Their specificity relies on the use of the “Tanche” olive variety harvested at full maturity and their slow spontaneous fermentation in 10% salt brine driven by yeast populations. This study aimed at investigating the benefit of inoculating autochthonous consortia to produce Nyons table olives by fermentation in 10% salt brine and in reduced salt conditions (8%). Two strategies were evaluated: inoculation with a defined autochthonous consortium and inoculation by spent brine backslopping. To define the consortium, yeasts were selected among 48 autochthonous isolates and key features included high halotolerance, low pectinolytic and proteolytic activities, however none had β-glucosidase activities. The consortium included eight yeast strains with distinct technological properties belonging to five dominant species, i.e. Citeromyces nyonsensis, Pichia membranifaciens, Wickerhamomyces anomalus, Zygotorulaspora mrakii and Candida atlantica. Fermentation trials were conducted over a year and compared by evaluating microbial community shifts (16S and ITS metagenetics) and volatile profiles (GC-MS). Regarding fermentations with the defined consortium, four out of five species implanted in early stages while one, Pichia membranifaciens, persisted and largely dominated by the end of the fermentation. Altogether, inoculation with the defined consortium did not disrupt microbial shifts compared to traditional fermentations although minor differences were observed in volatile profiles. The backslopping method yielded the highest impact on microbial populations and olive volatile profiles, with higher ester abundances at the end of fermentation. Finally, reduced salt in brine gave very promising results as no deleterious effects on microbial communities, volatile dynamics but also safety criteria of the olives were observed compared to traditional fermented olives.

Metabolomic and Transcriptional Profiling of Oleuropein Bioconversion into Hydroxytyrosol during Table Olive Fermentation by Lactiplantibacillus plantarum.

ABSTRACTThis study aims to elucidate the mechanisms responsible for the bioconversion of oleuropein into low-molecular-weight phenolic compounds in two selected Lactiplantibacillus plantarum strains, namely, C11C8 and F3.5, under stress brine conditions and at two different temperatures (16°C and 30°C). For this purpose, we adopted an experimental strategy that combined high-resolution mass spectrometry, in silico functional analysis of glycoside hydrolase family 1 (GH1)-encoding candidate genes, and gene expression studies. The oleuropein hydrolysis products and the underlying enzymatic steps were identified, and a novel putative bgl gene was detected, using seven strains belonging to the same species as controls. According to metabolomic analysis, a new intermediate compound (decarboxymethyl dialdehydic form of oleuropein aglycone) was revealed. In addition, strain C11C8 showed a decrease in the oleuropein content greater than that of the F3.5 strain (30% versus 15%) at a temperature of 16°C. The highest increase in hydroxytyrosol was depicted by strain C11C8 at a temperature of 30°C. PCR assays and sequencing analyses revealed that both strains possess bglH1, bglH2, and bglH3 genes. Furthermore, a reverse transcription-PCR (RT-PCR) assay showed that bglH3 is the only gene transcribed under all tested conditions, while bglH2 is switched off in strain C11C8 grown at cold temperatures, and no transcription was detected for the bglH1 gene. The bglH3 gene encodes a 6-phospho-β-glucosidase, suggesting how phospho-β-glucosidase activity could belong to the overall metabolic strategy undertaken by L. plantarum to survive in an environment poor in free sugars, like table olives.IMPORTANCE In the present study, a new candidate gene, bglH3, responsible for the β-glucosidase-positive phenotype in L. plantarum was detected, providing the basis for the future marker-assisted selection of L. plantarum starter strains with a β-glucosidase-positive phenotype. Furthermore, the ability of selected strains to hydrolyze oleuropein at low temperatures is important for application as starter cultures on an industrial scale.

Current Status, Recent Advances, and Main Challenges on Table Olive Fermentation: The Present Meets the Future

Table olives are among the most well-known fermented foods, being a vital part of the Mediterranean pyramid diet. They constitute a noteworthy economic factor for the producing countries since both their production and consumption are exponentially increasing year by year, worldwide. Despite its significance, olive’s processing is still craft based, not changed since antiquity, leading to the production of an unstable final product with potential risk concerns, especially related to deterioration. However, based on industrial needs and market demands for reproducible, safe, and healthy products, the modernization of olive fermentation processing is the most important challenge of the current decade. In this sense, the reduction of sodium content and more importantly the use of suitable starter cultures, exhibiting both technological and potential probiotic features, to drive the process may extremely contribute to this need. Prior, to achieve in this effort, the full understanding of table olive microbial ecology during fermentation, including an in-depth determination of microbiota presence and/or dominance and its functionality (genes responsible for metabolite production) that shape the sensorial characteristics of the final product, is a pre-requisite. The advent of meta-omics technology could provide a thorough study of this complex ecosystem, opening in parallel new insights in the field, such as the concept of microbial terroir. Herein, we provide an updated overview in the field of olive fermentation, pointing out some important challenges/perspectives that could be the key to the olive sector’s advancement and modernization.

New procedure to pre-select lactic acid bacteria able to control table-olive fermentation

Using 35 artisanal Calabrian table-olive brines, 153 strains of lactic acid bacteria were isolated. A screening procedure was designed, involving the following steps: 1) Gram stain, cellular morphology, homolactic fermentation, acidifying activity in MRS broth; 2) growth in olive paste with NaCl 11%; 3) increase of the antioxidant activity; 4) identification of the best strains. The proposed procedure allows an evaluation of the technological aptitude of lactic acid bacteria to ferment table olives excluding, cheaply and easily, all those strains definitely unable to act as starter. The use of olive paste as a screening medium allows the resistance of the new isolates against olive phenolic compounds to be tested. The selection of strains able to increase the antioxidant power of olive paste can improve the shelf-life of table olives.

Effects of Lactiplantibacillus plantarum 11 and Chloride Sodium on Biochemical and Microbiological Quality of Table Olives during Fermentation

Background: Table olives are nutritionally a complete food and considered as one of the oldest fermented products. This study aimed to evaluate the effect of Lactiplantibacillus plantarum 11 as a starter culture on the fermentation of table olives at two incubation temperatures 22 and 30 °C and different salt concentrations (0, 4, 8, and 12% m/v) of sodium chloride (NaCl). Methods: The fermentation of table olives was carried out according to the Spanish style. L. plantarum 11 was inoculated as a starter culture (106 Colony Forming Unit (CFU)/ml), and Listeria monocytogenes CECT 4032 was used as an indicator strain. Under the same experimental conditions, the fermentation of olives without the inoculation of starter culture was used as a control. Then, biochemical and microbiological quality of each experimental batch was tested. Results: Unlike the incubation temperature of 22 °C, the pH values ​​obtained in salted batches and incubated at 30 °C were all below the marketing limits for table olives. At the end of the process, the maximum load of yeasts and molds (>5 log CFU/ml) was recorded in the batches incubated at 22 °C. At 22 °C, Listeria was absent in inoculated fermenters at a concentration greater than or equal to 8% (w/v) of NaCl. However, at 30 °C, Listeria was not detected in treatment groups and in the control group with 12% NaCl. Conclusion: L. plantarum 11 could be potentially considered as a probiotic starter culture during the fermentation of black table olives.

Lactiplantibacillus plantarum Used as Single, Multiple, and Mixed Starter Combined with Candida boidinii for Table Olive Fermentations: Chemical, Textural, and Sensorial Characterization of Final Products

In this study, four different kinds of table olive fermentations belonging to Olea europaea L. Itrana cultivar were evaluated: A, spontaneous fermentation; B, fermentation with a single inoculum (Lactiplantibacillus plantarum B1); C, fermentation with multiple inoculum (L. plantarum B1 + L. plantarum B51 + L. plantarum B124, 1:1:1); and D, fermentation with mixed (bacterium + yeast) inoculum (L. plantarum B1 + Candida boidinii). This research focuses on the correlation between the different mixes of inoculations and their effect under the chemical, sensorial, and textural profiles in the final products (olives) for potential applications on table olive fermentation. During the fermentation, some specific parameters were monitored: chemical characterization of oil fraction (pigments, tocopherols, fatty acids, alkyl esters, and sterol composition), Texture Profile Analysis (TPA), determination of olive color, and sensory evaluation of the final products. The use of LAB starters (single and multiple inocula) compared to spontaneous process revealed a greater performance in preventing the spoilage process and in developing favorable physico-chemical conditions during the fermentation. In fact, the highest values of fatty acid alkyl esters were reached in spontaneous fermentation (~480 mg/kg in jar A). The presence of C. boidinii as inoculum in jar D was involved in table olive softening: the fermented olives showed the lowest values of the parameters related to consistence of fruit as hardness (~2300 g) and gumminess (~990 g) and high value of fatty acid methyl esters (~110 mg/kg).

A newly developed method based on surface physicochemical proprieties, for measuring the maturation level of olive fruit

The maturation process is an important determining factor to initiate the fermentation process in olive fruit. Level of maturation classically determined by the color of the olive fruit. In this study, we aim to develop a measurable criterion based on physicochemical properties and surface roughness of two important olive varieties grown in Morocco. The hydrophobic/hydrophilic properties and the electron donor-acceptor character of the olives were calculated. The obtained results showed a very clear change in the electron donor character and the surface roughness of the two studied varieties. This change depended on the degree of maturity and the nature of the cultivar. The electron donor character decreased (two times) with an increasing degree of maturity for the Arbequina, contrary to the Picholine variety, which showed a significant increase (4 times). Surface roughness increased 10 times for the Arbequina and decreased 5 times for the Picholine as maturity progressed from the green to the black stage. These findings may be useful in the management/prediction of the process of table olive fermentation.

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.

Behavior of Vibrio spp. in Table Olives.

The presence of Vibrio species in table olive fermentations has been confirmed by molecular biology techniques in recent studies. However, there has been no report of any foodborne outbreak caused by Vibrio due to the consumption of table olives, and their role as well as the environmental conditions allowing their survival in table olives has not been elucidated so far. The aims of this work were to model the behavior of an inoculated Vibrio cocktail in diverse table olive environments and study the possible behavior of an inoculated Vibrio cocktail in table olives. First, an in vitro study has been performed where the microbial behavior of a Vibrio cocktail was evaluated in a laboratory medium and in olive brines using predictive models at different NaCl concentrations (2–12%) and pH levels (4.0–9.0). Afterward, a challenge testing was done in lye-treated olives inoculated at the beginning of fermentation with the Vibrio cocktail for 22 days. The Vibrio cocktail inoculated in table olives has not been detected in olive brines during fermentation at different pH levels. However, it was observed that this microorganism in a laboratory medium could reach an optimal growth at pH 9 and 2% salt, without time of constant absorbance (tA), and the maximum absorbance value (yend) observed was at pH 8 and 2% salt conditions. The statistical analysis demonstrated that the effect of salt concentration was higher than pH for the kinetic growth parameters (μmax, tA, and yend). On the other hand, it was confirmed that no growth of the Vibrio cocktail on any sample was noticed in lye-treated olive fermentations. Thus, it was concluded that the presence of olive compounds (unknown) did not allow the development of Vibrio strains, so it is a very safety product as it has a natural antimicrobial compound, but the possibility that a native Vibrio sp. is able to acquire the capacity to adapt to this compound should be considered in further studies.

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.

Yeast dynamics in the black table olives processing using fermented brine as starter

Spontaneous fermentation with autochthonous yeasts is the most traditional method used for the production of black table olives, which requires minimal intervention. The aim of this study was to evaluate the effects of adding a brine starter, developed from one-year spontaneously fermented black table olives, on the microbiological, chemical, and sensory characteristics of naturally fermented Taggiasca table olives. The olives were fermented in brine containing 12% (w/v) NaCl with 0.6% (w/v) citric acid and inoculated with a selected brine starter containing primarily Pichia manshurica (87% predominance) and Saccharomyces cerevisiae (10% predominance). Brine and olives samples were analysed at the start of the experiment and after 3, 6, 9, and 12 months of fermentation. The P. manshurica and S. cerevisiae present in the brine starter drove the entire fermentation process, along with other autochthonous species that were not included in the starter, including Zygosaccharomyces mrakii. Spoilage microorganisms and undesired off-odours and off-flavours were not present at the end of fermentation. When compared to the control, the use of the brine starter increased the total yeast population in the brine during the first 6 months of fermentation, thus shortening the debittering time of the fruits and improving the taste complexity of the final product.