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This study delved into the evolution of fungal population during the fermentation of Spanish-style green table olives (Manzanilla cultivar), determining the influence of different factors such as fermentation matrix (brine or fruit) or the use of a lactic acid bacteria inoculum, on its distribution. The samples (n = 24) were directly obtained from industrial fermentation vessels with approximately 10.000 kg of fruits and 6.000 L of brines. Our findings showcased a synchronized uptick in lactic acid bacteria counts alongside fungi proliferation. Metataxonomic analysis of the Internal Transcribed Spacer (ITS) region unearthed noteworthy disparities across different fermentation time points (0, 24, and 83 days). Statistical analysis pinpointed two Amplicon Sequence Variants (ASV), Candida and Aureobasidium, as accountable for the observed variances among the different fermentation time samples. Notably, Candida exhibited a marked increase during 83 days of fermentation, opposite to Aureobasidium , which demonstrated a decline. Fungal biodiversity was slightly higher in brines than in fruits, whilst no effect of inoculation was noticed. At the onset of fermentation, prominently detected genera were also Mycosphaerella (19.82 %) and Apohysomyces (16.31 %), hitherto unreported in the context of table olive processing. However, their prevalence dwindled to nearly negligible levels from 24th day fermentation onwards (<2 %). On the contrary, they were replaced by the fermentative yeasts Saccharomyces and Isstachenkia. Results obtained in this work will be useful for designing new strategies for better control of table olive fermentations. • Metataxonomic analysis of ITS region was applied in Spanish-style olive fermentations. • Variances in yeast population were detected at 0, 24, and 83 days of fermentation. • Candida and Aureobasidium were the responsible genera for the statistical differences • No significant differences were observed in fermentation matrix or inoculation process. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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, a diverse genus belonging to the Enterobacteriaceae family appeared in a high relative abundance in pepper paste and garlic, and the 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 to table olive processing, as well as the presence of others which could compromise the stability of olive packaging. • Metataxonomic analysis has provided a deep knowledge of the microbiota present in olive dressing. • Pseudomonas and diverse non-assigned genus of Enterobacteriaceae predominated in aromatic herbs. • Lactobacillus , Weisella, and Pediococcus were the predominant genera found in many fermented dressing samples. • Salinibacter was the most representative genus found in samples of salt. • Olive dressing includes spoilage microorganisms that may alter the stability of table olive packaging. [ABSTRACT FROM AUTHOR]

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 10 cfu/mL) for all species assayed. The maximum death rate ( k max ) was 9.98, 51.37, 38.35 and 53.01 h −1 , while the time for 4 log 10 reductions ( 4D r ) 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 max . 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. [ABSTRACT FROM AUTHOR]

In the present study, a dose-response model was used to investigate the susceptibility (NIC) and resistance (MIC) of the lactic acid bacteria and yeast populations with respect to five chemical preservatives (fumaric and pyruvic acids, cinnamaldehyde, sodium metabisulphite and natamycin) with potential application in table olives. Results were compared with respect to potassium sorbate, a well-known preservative habitually used in olive packaging. Sodium metabisulphite was the most efficient preservative to control lactic acid bacteria growth (MIC, 50 ppm), followed by cinnamaldehyde (1060 ppm) while pyruvic acid required higher concentrations (3211 ppm). Natamycin (25 ppm) was highly efficient against yeasts, followed by cinnamaldehyde (125 ppm), potassium sorbate (553 ppm), sodium metabisulphite (772 ppm) and pyruvic acid (3038 ppm). Fumaric acid, in the range assayed (0–2000 ppm), did not show any inhibitory effect against these two microbial groups. This survey presents for the first time a comparative study of the efficiency of potential preservatives to control the growth of table olive related microorganisms. Further studies should be performed to validate their effects and interactions in the food matrix. [ABSTRACT FROM AUTHOR]

This work focuses on the persistence of the putative probiotic bacteria Lactobacillus pentosus TOMC- LAB2 on green Spanish-style Manzanilla olives according to different packaging conditions and storage temperatures. The lactic acid bacteria population decreased with time but the highest survival counts (and lowest yeasts) at the end of storage (8 months) were observed in plastic pouches under nitrogen atmosphere and glass jars with brine stored at 20°C. Molecular techniques showed a 100% presence of the putative probiotic bacteria in biofilms adhered to olive epidermis, while it was absent in PPB (plastic pouches with brine) and in olives stored at 7°C. No changes in NaCl, pH or combined acidity were observed during the storage except for a slight increase in titratable acidity at 20°C. The color of the fruits was stable but degraded at 20°C for olives in plastic pouches with brine. [ABSTRACT FROM AUTHOR]

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 10 CFU/g in 25% of cases) to a maximum of 5.86 log 10 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. • Metataxonomic approach of ITS region was applied in global commercial olive packages. • Half of total sequences obtained from all packages corresponded to the Pichia genus. • Potential pathogenic fungi were detected in high representation in some samples. • The presence of certain fungal taxa was influenced by the packaging conditions. [ABSTRACT FROM AUTHOR]

BACKGROUND Zinc chloride has been used previously as a preservative in directly brined olives with promising results. However, this is the first time that the effects of ZnCl2 addition (0-1 g L−1) on green Spanish-style table olive (cv. Manzanilla) packaging has been studied. RESULTS The presence of ZnCl2 affected the physico-chemical characteristics of the products; the presence of the Zn led to lower pH values (particularly just after packaging) and titratable and combined acidity values than the control but did not produce clear trends in the colour parameters. No Enterobacteriaceae were found in any of the treatments evaluated. At the highest ZnCl2 concentrations, the lactic acid bacteria were inhibited while, unexpectedly, its presence showed a lower effect than potassium sorbate against the yeast population. Regardless of the use of potassium sorbate or ZnCl2, the packages had a reduced microbial biodiversity because only Lactobacillus pentosus and Pichia galeiformis were found at the end of the shelf life. With respect to organoleptic characteristics, the presentations containing ZnCl2 were not differentiated from the traditional product. CONCLUSION Zinc chloride was less efficient than potassium sorbate as a yeast inhibitor in green Spanish-style olives, showing clear presentation style dependent behaviour for this property. Its presence produced significant changes in chemical parameters but scarcely affected colour or sensory characteristics.© 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND: Zinc chloride has been used previously as a preservative in directly brined olives with promising results. However, this is the first time that the effects of ZnCl2 addition (0–1 g L−1) on green Spanish-style table olive (cv. Manzanilla) packaging has been studied. RESULTS: The presence of ZnCl2 affected the physico-chemical characteristics of the products; the presence of the Zn led to lower pH values (particularly just after packaging) and titratable and combined acidity values than the control but did not produce clear trends in the colour parameters. No Enterobacteriaceae were found in any of the treatments evaluated. At the highest ZnCl2 concentrations, the lactic acid bacteria were inhibited while, unexpectedly, its presence showed a lower effect than potassium sorbate against the yeast population. Regardless of the use of potassium sorbate or ZnCl2, the packages had a reduced microbial biodiversity because only Lactobacillus pentosus and Pichia galeiformis were found at the end of the shelf life. With respect to organoleptic characteristics, the presentations containing ZnCl2 were not differentiated from the traditional product. CONCLUSION: Zinc chloride was less efficient than potassium sorbate as a yeast inhibitor in green Spanish-style olives, showing clear presentation style dependent behaviour for this property. Its presence produced significant changes in chemical parameters but scarcely affected colour or sensory characteristics. [ABSTRACT FROM AUTHOR]

After moisture, fat is the major constituent of table olives. However, scarce studies have been carried out to determine the influence of microorganisms and type of processing on the modification of their quality indexes. The present survey studies the influence of lipolytic ( Candida boidinii TOMC Y5 and Wickerhamomyces anomalus TOMC Y10) and nonlipolytic ( Debaryomyces etchellsii TOMC Y9 and Pichia galeiformis TOMC Y27) yeasts on the oil quality indexes of Manzanilla and Hojiblanca green fruits processed as directly brined and lye-treated table olives. Overall, the inocula scarcely used available sugars, except the lipolytic C. boidinii strain in lye-treated olives. Acetic acid production was limited in all conditions, except for the D. etchellsii strain in directly brined Manzanilla fruits. Ethanol formation was also reduced, although the W. anomalus (in both types of elaboration) and the C. boidinii (in lye-treated olives) strains produced significantly higher proportions. Apparently, changes in the oil quality indexes of processed olives were not related to the presence of yeasts, and hence, could have been caused by the endogenous activity of the fruits. A principal component analysis using the microbiological, physicochemical, and oil quality data supported this hypothesis, grouping treatments according to olive variety and type of elaboration, while segregation due to yeast inocula was not observed. [ABSTRACT FROM AUTHOR]

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. • Bacterial profiles of spoiled industrial table olive fermentation vessels was studied. • Butyric spoilage was mainly related to Enterococcus , Leuconostoc , and Atlantibacter. • Sulfidic spoilage was primary associated with Alkalibacterium. • Putrid spoilage was linked to Halolactobacillus and Alkalibacterium. • Propionibacterium , Bacillus or Pseudomonas were not detected in the spoiled vessels. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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, identification and quantification of fungi species in olive fermentation will facilitate the design of new strategies to improve the quality and safety of this fermented vegetable. [ABSTRACT FROM AUTHOR]

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. • Compositional analysis provided new insights of food metataxonomic data. • Low number of bacterial genera typifies green Spanish-style table olive processing. • Lactiplantibacillus became the predominant genera as the fermentation progressed. • Inoculation led to more homogenous bacterial populations at the end of the process. [ABSTRACT FROM AUTHOR]