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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]

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. [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]

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]

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. [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]