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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 mug mL-1. Many exhibited anti-inflammatory properties (reduction of pro-inflammatory cytokines TNF-alpha 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 mug 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.

Table olives are the most important fermented vegetable in the Mediterranean countries. In a continuous effort to improve the quality of this food, this work applies an experimental design with four variables to study the use of 1-methylcyclopropene (1-MCP, 2.85 µL/L) in combination with a light heat shock (65 °C, 5 min) and dimethyl dicarbonate (DMDC, 83 mg/L), during the processing of Spanish style and natural table olives. Fruits processed in the Spanish style exhibited higher moisture levels, luminance, and combined acidity. On the contrary, natural olives contained more sugars, had a firmer texture, and displayed a greener colour. Our results show that applying 1-MCP, heat shock, and DMDC can positively impact the processing of Spanish style and natural table olives and influence their physicochemical characteristics. Adding DMDC to the initial brine caused colour degradation but lowered the pH. Using 1-MCP with a light heat shock improved the sugar content and luminance of olives and increased the titratable acidity in the brine. Applying 1-MCP to the Spanish style enhanced the luminosity (brightness) and, when combined with DMDC, led to the lowest pH levels. Further research at an industrial scale is necessary to validate these results and study their influence on microorganisms. These treatments could have applications in developing new table olive processing methods that allow for more predictable products.

Aquaculture is becoming a strategic sector for many national economies to supply the increasing demand for fish from consumers. Fish culture conditions and processing operations can lead to an increase in microbial contamination of farmed fish that may shorten the shelf-life of fish products and byproducts, and ready-to-eat fishery products. The objective of this study was to evaluate the hygienic-sanitary status of water, environment, and processing of fresh-farmed rainbow trout (Oncorhynchus mykiss) fillets produced in a local fish farm in Andalusia, Spain. To achieve this, a longitudinal study was carried out by collecting environmental (air and food-contact surfaces), water from fish ponds, and rainbow trout samples. Thereby, seven sampling visits were performed between February 2021 and July 2022, where foodborne pathogens and spoilage microorganisms, together with physicochemical parameters, were analysed in the collected samples. Further, microbial identification of microbiota was achieved through a culture-dependent technique using blast analysis of 16S RNA gene sequencing. The results showed that Listeria monocytogenes and Salmonella were not detected in the analysed samples. Regarding the hygienic-sanitary status of the fish farm, the slaughtering bath, the eviscerating machine and the outlet water from fish ponds presented the highest counts of coliforms, Enterobacteriaceae, and Aerobic Mesophilic Bacteria. Staphylococcus aureus and sulphite-reducing Clostridium were identified in the conveyor belts, fish flesh, and viscera. The 16S RNA identification confirmed the presence of viable spoilage bacteria such as Citrobacter gillenii, Macrococcus caseolyticus, Hafnia paralvei, Lactococcus lactis, Lactococcus cremoris, Klebsiella, Escherichia coli, Morganella morganii, and Shewanella. Three of these genera (Citrobacter, Hafnia, and Pseudomo

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 log CFU/g in 25% of cases) to a maximum of 5.86 log 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.

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.

Aloreña de Málaga is a table olive especially characterised by its natural freshness and short shelf-life. In this work, we applied a metataxonomic approach to unravel the microbial diversity of bacterial and fungi populations through the shelf-life of traditionally packed Aloreña de Málaga. A significant increase in lactic acid bacteria and mesophilic aerobic populations was observed during shelf-life, reaching the maximum population levels (4–5 log10 CFU) at the end of the study (260 days). On the contrary, a rapid reduction in yeast and mould populations was reported. The use of a metataxonomic analysis based on the amplification of 16S (bacteria) and internal transcribed spacer (ITS) region (fungi) regions revealed a low diversity for both microbial groups. Lactiplantibacillus (65.05 ± 8.65% in brine vs. 58.70 ± 15.70% in fruit), Pediococcus (28.17 ± 7.36% in brine vs. 27.20 ± 15.95% in fruit), and Celerinatantimonas (4.64 ± 1.08% in brine vs. 11.82 ± 18.17% in fruit) were the main genera found among bacteria, and an increase in Lactiplantibacillus and a reduction in Celerinatantimonas populations during the shelf-life were observed. On the other hand, Citeromyces was the dominant fungi genus (54.11 ± 2.00% in brine vs. 50.91 ± 16.14% in fruit), followed by Candida (8.80 ± 2.57% in brine vs. 12.32 ± 8.61% in fruit) and Penicillium (6.48 ± 1.87% vs. 8.48 ± 4.43% in fruit). No food-borne pathogen genera were detected in any of the samples analysed, indicating the high level of food safety found in this ready-to-eat fermented vegetable. Data obtained in this work will help in the design of new strategies for the control of microbial populations during the shelf-life of Aloreña de Málaga.

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.

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 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.

Vida de mercado de aceitunas tradicionales Aloreña de Málaga aliñadas basado en la apariencia del envase y características del fruto. Pocos estudios han sido llevados a cabo para determinar la vida de mercado de aceitunas de mesa Aloreña de Málaga envasadas desde un punto de vista físico-químico, microbiológico y sensorial. Este trabajo muestra, cómo, bajo las actuales condiciones de envasado, los productos comerciales están libres de enterobacterias, la población inicial de levaduras se inhibe progresivamente, mientras que las bacterias ácido lácticas son las únicas que se desarrollan durante el envasado. Entre las características físico-químicas, el pH descendió, se formó ácido láctico mientras que el manitol y el ácido cítrico fueron consumidos. Estos cambios dieron lugar a una progresiva degradación de la textura y a una depreciación del color verde de los frutos. El análisis multivariante realizado mostró que los envasados entre los 6 y 42 días de vida de mercado fueron los mejores valorados por los panelistas, mientras que después de los 74 días, el producto envasado fue perdiendo aceptabilidad, lo cual fue especialmente evidente tras 131 días de envasado (la predisposición de compra fue solo del 50%). Una completa estabilización microbiológica del producto requeriría del uso de conservantes alternativos, ya que el tratamiento térmico no es muy conveniente para este tipo de elaboración., Few studies have been carried out to determine the shelf-life of the Aloreña de Málaga table olive packaging from a physicochemical, microbiological and sensorial point of view. This study showed that under the current packaging conditions, commercial products were free from Enterobacteriaceae, the initial yeast population was progressively inhibited, and only lactic acid bacteria grew during shelf-life. Among the physicochemical characteristics, pH decreased, lactic acid was formed while citric acid and mannitol were consumed. These changes resulted in gradual olive texture degradation and green color fading during packaging. A multivariate analysis showed that the packaged olives with storage time between 6 and 42 days enjoyed the highest acceptance; while after the 74th day, they were progressively losing acceptability, which was mainly evident at the 131st day of packaging (willingness-to-buy attribute was reduced to 50%). A complete microbiological stabilization would require the use of alternative preservatives since thermal treatment is not convenient for this type of olive speciality.

In this work, we have used a metataxonomic analysis to study the bacterial biodiversity in three types of commercial dressing and ingredients habitually used in table olive packaging. The bioinformatics analysis showed that salt samples (marine and spring) had the highest biodiversity indexes, followed by the aromatic herbs (thyme, oregano, and fennel) and finally by fermented dressing samples (garlic, pepper, and pepper paste). In salt samples, a significant high presence of Cytophagia, Sphingobacteria, Deltaproteobacteria and Halobacteria classes was observed. On the contrary, the presence of Bacilli class in fermented dressing samples was significantly higher compared to the other samples. Finally, the abundance of Gammaproteobacteria was statistically higher in the samples of herbs. At genus or family taxonomic level, Pseudomonas and diverse non-assigned genus of the Enterobacteriaceae family predominated in many samples of herbs. In fermented dressing samples, diverse genus belonging to the Enterobacteriaceae family appeared in a high relative abundance in pepper paste and garlic, and presence of Vibrio sp. was detected in pepper. However, Lactobacillus, Weisella, and Pediococcus were the predominant genera found in many of these fermented samples. Salinibacter was the most representative genus in both types of salt samples. Multivariate analysis (PCA) of the metataxonomic data sets discriminated well the samples belonging to the three different groups, showing a high proportion of sequences shared by samples-group. This survey reports for the first time the presence of new bacterial species not previously related with table olive processing, as well as the presence of others which could compromise the stability of olive packaging.

The work assays the use of various concentrations of ZnCl2 (0.0–0.1%, w/v) in packaged natural black Manzanilla table olives. The transformations were followed for 4 months. The presence of Zn modified the leaching of total sugars (sucrose, glucose, fructose, and mannitol) into the brine, which decreased as the ZnCl2 content increased. Over the study, sucrose and glucose were exhausted while fructose, although consumed, left some final residues and the use of mannitol was limited. Titratable acidity was always gradually formed causing the subsequent pH decrease, which stabilized at ≈3.5. Acetic and mainly lactic acid were also formed during the assay, reaching the highest level of lactic acid in the 0.050% ZnCl2 treatment, followed by the Control. The acids were formed by lactic acid bacteria (LAB) (Lactobacillus pentosus, 39%, and Lactobacillus plantarum, 61%). However, the most outstanding Zn effect was found on the olive sensory characteristics: its presence markedly reduced the bitter notes, increased the overall appreciation, and the treatment containing 0.075% ZnCl2 had the highest scores in hardness, crunchiness, and overall appreciation. Therefore, the addition of ZnCl2 into packaged natural table olives may lead to healthy products with desirable sensory characteristics which, in turn, could promote consumption.

This study presents an approach to determine the survival of diverse foodborne pathogens (Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica) in three Aloreña de Málaga table olive commercial presentations (fresh green, traditional, and cured olives). The microbial survival in this green natural table olive speciality was fit using a log-linear regression model implemented in GInaFIT. The contents of sugars, phenolic compounds, additives, salt, pH, and levels of autochthonous microorganisms differed among presentations and affected the survival of microorganisms. The inoculated initial populations of pathogens (7-8 log CFU/mL brine) decreased rapidly and, 48 h after inoculation, their counts were always below the detection limit (<1.3 log CFU/mL), except for S. aureus in the fresh green presentation which was ~ 5.0 log CFU/mL. The highest maximum death rates (k) and lowest periods for 4 log reductions (4Dr) were observed in cured olives but decreased and increased, respectively, from the traditional to the fresh green presentations. L. monocytogenes and S. aureus were the most resistant species. The multivariate analysis showed that high concentrations of compounds released from the olives (sugars and phenols) were positively associated to 4Dr and negatively to k. Conversely, the presence of preservatives reduced 4Dr. This study, executed in commercial products, pointed out that packaged table olives are adverse habitats for foodborne pathogens with their effects being presentation dependent. The survival of S. aureus was particularly long in green fresh Aloreña de Málaga table olives packaged without preservatives; therefore, its changes in this presentation still requires further in-deep research.

Table olives are one of the most representatives and consumed fermented vegetables in Mediterranean countries. However, there is an evident lack of standardization of production processes and HACCP systems thus implying the need of establishing decision-making tools allowing their commercialization and shelf-life extension. The present work aims at developing a decision-making scoring system by means of a probabilistic assessment to standardize production process of Aloreña de Málaga table olives based on the identification of potential hazards or deficiencies in hygienic processes for the subsequent implementation of corrective measures. A total of 658 microbiological and physico-chemical data were collected over three consecutive olive campaigns (2014–2016) to measure the variability and relative importance of each elaboration step on total hygienic quality and product safety. Three representative companies were visited to collect samples from food-contact surfaces, olive fruits, brines, air environment, olive dressings, water tanks, and finished/packaged products. A probabilistic assessment was done based on the establishment of Performance Hygiene and Safety Scores (PHSS 0–100%) through a standardized system for evaluating product acceptability. The mean value of the global PHSS for the Aloreña de Málaga table olives processing (PHHSFTOT) was 64.82% (90th CI: 52.78–76.39%) indicating the high variability among facilities in the evaluated processing steps on final product quality and safety. Washing and cracking, and selection and addition of olive dressings were detected as the most deficient ones in relation to PHSSFi values (p < 0.05) (mean = 53.02 and 56.62%, respectively). The relative contribution of each processing step was quantified by different experts (n = 25) from the Aloreña de Málaga table olive sector through a weighted PHSS (PHSSw). The mean value of PHSSw was 65.53% (90th CI: 53.12–77.52%). The final processing steps obtained higher values for PH

Preserving the highly appreciated natural freshness of Aloreña de Málaga table olives and preventing their progressive darkening during processing is a major challenge. In this work, heat-shocked (60°C, 5 min) fruits were processed according to the three denominations referred to in the Protected Designation of Origen (cured, fresh green, and traditional) and their characteristics compared with those that followed the habitual industrial process (controls). The results revealed that the effects of the heat treatment on the evolution of pH, titratable acidity, salt, sugar, organic acid, ethanol content, texture, and color of fruits as well as on microbial populations (yeasts and lactic acid bacteria) were slight in the case of the fresh green and cured presentations. However, the differences between heat-shocked and its control were remarkable in the traditional process. Notably, the heat treatment favored lactic acid fermentation, retention of the green appearance of the fruits, stability during packaging, and led to the highest sensory evaluation. The metagenomic analysis carried out at the end of the fermentation revealed the presence in all samples of three genera (Lactobacillus, Pediococcus, and Celerinatantimonas) which encompassed most of the sequences. The number of Lactobacillus sequences was statistically higher (p ≥ 0.05) in the case of traditional heat-shocked fruits than in its control.

This work studies the inoculation conditions for allowing the survival/predominance of a potential probiotic strain (Lactobacillus pentosus TOMC-LAB2) when used as a starter culture in large-scale fermentations of green Spanish-style olives. The study was performed in two successive seasons (2011/2012 and 2012/2013), using about 150 tons of olives. Inoculation immediately after brining (to prevent wild initial microbiota growth) followed by re-inoculation 24 h later (to improve competitiveness) was essential for inoculum predominance. Processing early in the season (September) showed a favorable effect on fermentation and strain predominance on olives (particularly when using acidified brines containing 25 L HCl/vessel) but caused the disappearance of the target strain from both brines and olives during the storage phase. On the contrary, processing in October slightly reduced the target strain predominance on olives (70–90%) but allowed longer survival. The type of inoculum used (laboratory vs. industry pre-adapted) never had significant effects. Thus, this investigation discloses key issues for the survival and predominance of starter cultures in large-scale industrial fermentations of green Spanish-style olives. Results can be of interest for producing probiotic table olives and open new research challenges on the causes of inoculum vanishing during the storage phase.

This study uses a mathematical approach to assessing the inhibitory effect of Zn2 + (0–10 mM, obtained from ZnCl2 and ZnSO4) in presence of NaCl (0–8%) and hydroxytyrosol (0–2588 mg/L), on a yeast cocktail formed by species Pichia galeiformis, Pichia kudriavzevii, Pichia manshurica and Candida thaimueangensis obtained from spoilt green olive packages. The logistic/probabilistic models were built in laboratory medium using a total of 1980 responses (1188 for NaCl and 792 for hydroxytyrosol). ZnCl2 showed significantly higher inhibitory effect than ZnSO4 in the presence of both NaCl (p < 0.033) and hydroxytyrosol (p < 0.009). NaCl did not interfere the effect of Zn2 + while hydroxytyrosol, at high levels, had a slight antagonistic effect. According to models, Zn2 + inhibits (p = 0.01) the yeast cocktail in the range 4.5–5.0 mM for ZnCl2, or 8.5–9.5 mM for ZnSO4. Therefore, this work confirms the fungicidal activity of zinc compounds (mainly ZnCl2) in synthetic medium, and also shows that the loss of zinc effectiveness in real green Spanish-style olive packaging is not due to the presence of NaCl or hydroxytyrosol, two of the most abundant chemical compounds in the product.

This study uses an ¿omics¿ approach to evaluate the bacterial biodiversity changes during fermentation process of natural green cracked Aloreña de Málaga table olives, from raw material to fermented fruit. For this purpose, two industries separated by almost 20 km in Guadalhorce Valley (Málaga, Spain) were analysed for obtaining both brines and fruit samples at different moments of fermentation (0, 7, 30 and 120 days). Physicochemical and microbial counts during fermentation showed the typical evolution of this type of processes, apparently dominated by yeasts. However, high-throughput barcoded pyrosequencing analysis of V2¿V3 hypervariable region of the bacterial 16S rRNA gene showed at 97% identity the presence of 131 bacterial genera included in 357 operational taxonomic units, not detected by the conventional approach. The bacterial biodiversity was clearly higher in the olives at the moment of reception in the industry and during the first days of fermentation, while decreased considerably as elapse the fermentation process. The presence of Enterobacteriaceae and Lactobacillaceae species was scarce during the four months of study. On the contrary, the most important genus at the end of fermentation was Celerinatantimonas in both brine (95.3% of frequency) and fruit (89.4%) samples, while the presence of well-known spoilage microorganisms (Pseudomonas and Propionibacterium) and halophilic bacteria (Modestobacter, Rhodovibrio, Salinibacter) was also common during the course of fermentation. Among the most important bacterial pathogens related to food, only Staphylococcus genus was found at lowfrequencies (b0.02% of total sequences). Results showthe need of this type of studies to enhance our knowledge of the microbiology of table olive fermentations. It is also necessary to determine the role played by these species not previously detected in table olives on the quality and safety of this fermented vegetable.

The present study focused on investigating a peculiar spoilage of traditional Aloreña de Málaga table olives characterized by the formation of whitish and soft regions on the olive surface. To determine its causes, the main microbiological and physicochemical changes in 50 commercial packages were monitored until the alteration in appearance (63 days). Colour and firmness of fruits deteriorated progressively in the packaged olives during storage. Sugar in brines (7.5 g/L) remained stable during the first month and then decreased gradually (5.0 g/L) while, in parallel, the lactic acid bacteria population and titratable acidity increased. After two months of storage, evidence of spoilage was noticed, coinciding with the maximal lactic acid bacteria populations in fruit (>6.5 log10 CFU/g). The spoilage affected ¿25% of the fruits within packages. The microbial species detected in the product after packaging were Enterobacter gergoviae and Lactobacillus plantarum among bacteria, and Candida tropicalis, Candida parapsilosis, and Lodderomyces elongisporus among the yeasts while Lactobacillus pentosus was the dominant species in the spoilt packages. A specific biotype of L. pentosus was only detected in the damaged fruits. Further studies will be made to confirm the association between the spoilage and the presence of this L. pentosus biotype.

This work reports the survival (challenge tests) of foodborne pathogen species (Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica) in Aloreña de Málaga table olive brines. The inhibitions were fit using a log-linear model with tail implemented in GInaFIT excel software. The olive brine had a considerable inhibitory effect on the pathogens. The residual (final) populations (Fp) after 24 h was below detection limit (<1.30 log cfu/mL) for all species assayed. The maximum death rate (k) was 9.98, 51.37, 38.35 and 53.01 h, while the time for 4 log reductions (4D) was 0.96, 0.36, 0.36 and 0.24 h for E. coli, S. aureus, L. monocytogenes and S. enterica, respectively. Brine dilutions increased Fp and 4Dr, while decreased k. A cluster analysis showed that E. coli had an overall quite different behaviour being the most resistant species, but the others bacteria behaved similarly, especially S. aureus and S. enterica. Partial Least Squares regression showed that the most influential phenols on microbial survival were EDA (dialdehydic form of decarboxymethyl elenolic acid), HyEDA (EDA linked to hydroxytyrosol), hydroxytyrosol 4-glucoside, tyrosol, and oleoside 11-methyl ester. Results confirm the adverse habitats of table olives for foodborne pathogenic microorganisms.

A probabilistic/logistic model, based on binary data (growth/no growth), was used to assess the effects of sodium metabisulphite (SM) and cinnamaldehyde (CIN; 0-1000 mg/L) against the main microbial groups found in table olive environment [lactic acid bacteria (LAB), yeasts, and Enterobacteriaceae], according to pH (range 3.5-5.0), and type of acidifying agent (HCl or pyruvic acid). The inhibitory effect of SM depended on the pH while that of CIN was scarcely influenced by it (except for LAB). LAB were more sensitive to SM, while yeasts were to CIN. The use of pyruvic acid for correction of pH always produced a reduction (compared to HCl) of the inhibitory power of both preservatives. The in silico models for HCl showed that, at pH 4.0, and growth probability 0.01, the LAB population might be inhibited by the presence in the medium of 150 mg/L SM or 1000 mg/L CIN, while in the case of yeasts, 450 mg/L SM, or 150 mg/L CIN are required. No growth of Enterobacteriaceae was observed at this (or lower) pH level. The results obtained may contribute to the stabilization of non-thermally treated table olive packaging.

Nowadays, our knowledge of the fungal biodiversity in fermented vegetables is limited although these microorganisms could have a great influence on the quality and safety of this kind of food. This work uses a metagenetic approach to obtain basic knowledge of the fungal community ecology during the course of fermentation of natural Aloreña de Málaga table olives, from reception of raw material to edible fruits. For this purpose, samples of brines and fruits were collected from two industries in Guadalhorce Valley (Málaga, Spain) at different moments of fermentation (0, 7, 30 and 120 days). The physicochemical and microbial counts performed during fermentation showed the typical evolution of this type of processes, mainly dominated by yeasts in apparent absence of Enterobacteriaceae and Lactobacillaceae. High-throughput barcoded pyrosequencing analysis of ITS1-5.8S-ITS2 region showed a low biodiversity of the fungal community, with the presence at 97% identity of 29 different fungal genera included in 105 operational taxonomic units (OTUs). The most important genera in the raw material at the moment of reception in the industry were Penicillium, Cladosporium, Malassezia, and Candida, whilst after 4 months of fermentation in brines Zygotorulaspora and Pichia were predominant, whereas in fruits were Candida, Penicillium, Debaryomyces and Saccharomyces. The fungal genera Penicillium, Pichia, and Zygotorulaspora were shared among the three types of substrates during all the course of fermentation, representing the core fungal population for this table olive specialty. A phylogenetic analysis of the ITS sequences allowed a more accurate assignment of diverse OTUs to Pichia manshurica, Candida parapsilosis/C. tropicalis, Candida diddensiae, and Citeromyces nyonensis clades. This study highlights the existence of a complex fungal consortium in olive fermentations including phytopathogenic, saprofitic, spoilage and fermentative genera. Insights into the ecology, identificati