Suchergebnisse

13 Páginas.-- 2 Tablas.-- 5 Figuras ; 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 research has received funding from the Spanish Government (Project TOBE RTI2018-100883-B-I00: www.tobeproject.csic.es). ; Peer reviewed

Yeasts are unicellular eukaryotic microorganisms with a great importance in the elaboration on many foods and beverages. In the last years, researches have focused their attention to determine the favourable effects that these microorganisms could provide to table olive processing. In this context, the present study assesses, at laboratory scale, the potential technological (resistance to salt, lipase, esterase and β-glucosidase activities) and probiotic (phytase activity, survival to gastric and pancreatic digestions) features of 12 yeast strains originally isolated from Greek natural black table olive fermentations. The multivariate classification analysis carried out with all information obtained (a total of 336 quantitative input data), revealed that the most promising strains (clearly discriminated from the rest of isolates) were Pichia guilliermondii Y16 (which showed overall the highest resistance to salt and simulated digestions) and Wickerhamomyces anomalus Y18 (with the overall highest technological enzymatic activities), while the rest of strains were grouped together in two clearly differentiated clusters. Thus, this work opens the possibility for the evaluation of these two selected yeasts as multifunctional starters, alone or in combination with lactic acid bacteria, in real table olive fermentations. © 2014 Elsevier Ltd. ; The research leading to these results has received funding from the Spanish Government (Ramón y Cajal program), and the Junta de Andalucía (through financial support to group AGR-125). S. Bonatsou and F.N. Arroyo-López wish to express thanks to the European Erasmus program and Ramón y Cajal postdoctoral research contract (Spanish Government), respectively. ; Peer Reviewed

36 Páginas.-- 8 Figuras.-- 1 Tabla ; This work relates native lactic acid bacteria (LAB) (Lactobacillus pentosus LPG1, L. pentosus Lp13, and Lactobacillus plantarum Lpl15) and yeast (Wickerhamomyces anomalus Y12) starters to the volatile components (VOCs) produced in green Spanish-style table olives. For this aim, the VOC profile was considered as compositional data (CoDa). The CoDa analysis generated new information on the relationship among inocula and VOCs through the tetrahedral plot, CoDa-biplot, variation array matrix, and CoDa dendrogram. The ilr (which includes pivot) coordinates (Euclidean space) from VOCs produced more reliable starters' clustering than the original data. The potential VOC markers, identified by a test based on the pairwise comparison of the logratio variation arrays from the whole data set and the individual groups, were (starters in the parenthesis): 2-phenylethyl acetate (LPG1, Y12, Y12 + LAB), methanol (Lpl15), cis-2-penten-1-ol (LPG1, Y12, Y12 + LAB), 2-methyl-3-hexanol (LPG1, Y12), U (non-identified) C (m/z 83-112-97) (Y12) and UF (m/z 95-154-110) (LPG1, Y12 + LAB). Besides, some VOCs were partial/totally inhibited by specific starters: 2-methyl-1-propanol (Lp13, Y12 + LAB), 2-phenyl ethanol (Lp13), furfuryl methyl ether (Y12 + LAB), purpurocatechol (Y12, Y12 + LAB), 4-ethyl guaiacol (Lp13, Lpl15), 4-ethyl phenol (Lpl15), 5-tert-butylpyrogallol (Lp13, Lpl15), and UE (m/z 111–198) (Lp13). A better understanding of the relationship between starters and their VOC may facilitate modelling the flavour and quality of Spanish-style green table olive fermentations. ; The research was funded by the Spanish Government (Project OliFilm AGL-2013-48300-R: www.olifilm.science.com.es) A-BC thanks the Spanish Ministry of Economy and Competitiveness for their FPI grant. ; Peer reviewed

12 Páginas.-- 7 Figuras.-- 1 Tabla ; 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 work was supported by the European Union (Probiolives, contract 243471), Spanish Government (project AGL2010-15494/ALI, partially financed by European regional development funds, ERDF), and Junta de Andalucía (through financial support to group AGR-125). JB-G and FA-L wish to thank the CSIC and the Spanish government for their research contracts, respectively. ; Peer reviewed

This paper describes the dominant bacterial species metabolically active through the industrial production of Spanish-style Manzanilla and Gordal olives. For this purpose, samples (brines and fruits) obtained at 0, 15, and 90 fermentation days were analyzed by a culture-independent approach to determine viable cells by reverse transcription of RNA and further PCR-DGGE analysis, detecting at least 7 different species. Vibrio vulnificus, Lactobacillus plantarum group, and Lactobacillus parafarraginis were present in samples from both cultivars; Lactobacillus sanfranciscensis and Halolactobacillus halophilus were detected only in Gordal samples, while Staphylococcus sp. was exclusively found at the onset of Manzanilla fermentations. Physicochemical data showed a typical fermentation profile while scanning electron microscopy confirmed the in situ biofilm formation on the olive epidermis. Different Bacillus, Staphylococcus, and Enterococcus species, not detected during the fermentation process, were also found in the solid marine salt used by the industry for preparation of brines. Elucidation of these non-lactic acid bacteria species role during fermentation is then an appealingly challenge, particularly regarding safety issues. ; The research has received funding from the Spanish Government (Project OliFilm AGL-2013-48300-R: www.olifilm.science.com.es). AB-C and FA-L wish to express thanks to the Spanish Government for their pre-doctoral fellowship and postdoctoral research contract (Ramón y Cajal), respectively. ; Peer Reviewed

© 2015, Springer International Publishing Switzerland. This work examines the formation of poly-microbial communities adhered to the epidermis of natural green Gordal olives and the application of different methodologies for recovery and counting of the microorganisms embedded in olive biofilms. The fermentation process was physicochemical and microbiologically monitored for 90 days, at which, formation of true biofilms on the skin of fermented fruits was confirmed by scanning electron microscopy. Then, samples of olives were taken and treated with sonication, enzymes, mechanical homogenization with stomacher and ultrasonic bath for biofilm disaggregation. The use of the stomacher for 1 min was the most effective treatment to release the lactic acid bacteria (6.6 log 10 cfu g −1 ), whereas sonication for 5 min was the most efficient method for quantification of yeasts (up to 3.5 log 10 cfu g −1 ). Molecular identification of isolates obtained from natural Gordal olive biofilms revealed that Lactobacillus pentosus was the only species found among lactic acid bacteria, while Pichia membranifaciens was the dominant yeast species, with higher counts obtained for the bacteria. ; The research leading to these results has received funding from the Spanish Government (project OliFilm AGL-2013-48300-R: www.olifilm.science.com.es), and the Junta de Andalucía (through financial support to project P11-AGR-7051 and group AGR-125). A. Benítez and F.N. Arroyo-López wish to express thanks to the Spanish Government for their pre-doctoral fellowship and postdoctoral research contract (Ramón y Cajal), respectively. V. Romero-Gil would like also to thank the AgriFood Campus of International Excellence (ceiA3), Bank of Santander, Spanish Government and 'Aloreña de Málaga' Olive Manufacturing Association for her pre-doctoral fellowship (training program of Ph.D. in companies). ; Peer Reviewed

12 Páginas ; 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 research was funded by the Spanish Government through the TOBE project (RTI2018-100883-B-I00). ; Peer reviewed

11 Páginas; 5 Tablas; 6 Figuras ; 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. ; The research leading to these results has received funding from Junta de Andalucía Government through the PrediAlo project (AGR-7755: www.predialo.science.com.es) and FEDER European funds. FA-L wishes to express thanks to the Spanish government for his RyC postdoctoral research contract while VR-G would like to thank ceiA3, Spanish Government, Bank of Santander, IG-CSIC and 'Aloreña de Málaga' Olive Manufacturing Association for her predoctoral fellowship. Authors express their gratitude to Dr. Borja Sánchez (IPLA-CSIC) for his invaluable help in the metagenomic analysis. ; Peer reviewed

31 Páginas; 4 Figuras ; Directly brined black table olives of Bosana variety are a traditional food product of Sardinia island (Italy), spontaneously fermented by yeasts among other microorganisms. However, as far as we know, the identification, biotechnological and probiotic potential of this yeast community has not been investigated yet. In this work, a total of 72 yeast isolates previously obtained from Bosana olive brines were first genotyped by Random Amplified Polymorphic DNA (RAPD-PCR) analysis with primer M13, and then identified by sequencing of D1/D2 domains of rDNA 26S gene. The dominant species were Wickerhamomyces anomalus and Nakazawaea molendini-olei, albeit Candida diddensiae, Candida boidinii, Zygotorulaspora mrakii, and Saccharomyces cerevisiae were also present in lower proportions. For the different biotypes of yeasts obtained, the multivariate analysis of their technological (esterase, lipase and β-glucosidase activities, growth in presence of oleuropein, resistance and susceptibility to NaCl) and probiotic (removal of cholesterol, gastric and pancreatic digestions, biofilms assays alone and in co-culture with Lactobacillus pentosus) features, showed that W. anomalus Wa1 exhibited the best technological characteristics, while S. cerevisiae Sc24 and C. boidinii Cb60 showed promising probiotic features. Therefore, they may have potential application as multifunctional starters, alone or in combination with lactic acid bacteria, during olive processing, albeit further studies are necessary to validate these results. ; The research leading to these results has received funding from the Spanish Government (Project OliFilm AGL-2013-48300-R: www.olifilm.science.com.es), and Junta de Andalucía (through financial support to projects P11-AGR-7051, P11-AGR-7755, and groups AGR-125 and BIO-160). CP gratefully acknowledges Sardinia regional Government for the financial support of her PhD scholarship (POR Sardegna FSE OP European Social Fund2007/2013 – Axis IV Human Resources, Objective 1.3, Line of activity 1.3.1) and the Foundation of Sardinia (Prat. 2013.1364) for financial support. FNAL wish to express thanks to the Spanish Government for his postdoctoral research contract (Ramón y Cajal). Authors express their gratitude to Dr. Antonio Garrido Fernández for his constructive comments and suggestions with drafting of manuscript. CP gratetfully acknowledges Dr Ivo Pinna for kindly provided support of her thesis manuscript. ; Peer reviewed

14 Páginas, 14 Tablas, 5 Figuras ; Candida boidinii is an Ascomycota yeast with important biotechnological applications. In this paper we present the genome sequencing and annotation of eight strains of this species isolated from human activities and wild environments. The produced assemblies revealed several strain specific features in terms of genomic GC content (ranging from 30.9 to 32.7%), genome size (comprised between 18,791,129 and 19,169,086 bp) and total number of protein coding genes (ranging from 5819 to 5998), with putative assignation to their general KOG functional categories. The obtained data underlined the presence of two different groups for this species. The results reported herein provide new insights into the plasticity of the genome of this yeast species and represent a starting point for further studies in view of its biotechnological applications ; The research leading to these results has received funding from the Spanish Government (project OliFilm AGL-2013-48300-R: www.olifilm.science.com.es), and Junta de Andalucía (through financial support to project P11-AGR-7051 and groups AGR-125 and BIO-160). ABC and FNAL wish to express thanks to the Spanish Government for their pre-doctoral fellowship and postdoctoral research contract (Ramón y Cajal), respectively, while BCD is a recipient of a pre-doctoral grant from Junta de Andalucía. CP gratefully acknowledges Sardinia regional Government for the financial supporter of her PhD scholarship (POR Sardegna FSE OP European Social Fund 2007/2013 – Axis IV Human Resources, Objective 1.3, Line of activity 1.3.1) and the Foundation of Sardinia (Prat. 2013.1364) for financial support. ; Peer reviewed

3 Páginas.-- 1 Tabla ; The draft genome sequences of five Lactobacillus pentosus strains isolated from biofilms on the skin of green table olives are presented here. These genome sequences will assist in revealing the potential probiotic properties of these strains, as the intake of fermented olives implicates the passage of millions of Lactobacillus spp. throughout a consumer's gastrointestinal tract. ; This work was funded by the Junta de Andalucía through the P11-AGR-7051 project and by the Spanish Government through MICIIN projects AGL2013-48300-R/ALI (OliFilm project: www.olifilm.science.com.es) and BFU2016-77297-P. B.C.-D. was the beneficiary of a PIF grant from the Junta de Andalucía, and A.B.-C. was supported by the MICIIN through a PIF grant. We thank C3UPO for the high-performance computing (HPC) support. ; Peer reviewed

13 Páginas.-- 2 Figuras.-- 6 Tablas ; 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 (t), and the maximum absorbance value (y) 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 (μ, t, and y). 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. ; The research leading to these results has received funding from the Spanish government through the TOBE project (RTI2018-100883-B-I00) and Council of Economy, Knowledge, Business and University of Junta de Andalucía (Project AT 2017-5686, Project AT 2017-5503) and FEDER European funds.

Here, we report the genome sequences of six Lactobacillus pentosus strains isolated from traditional noninoculated Spanish-style green table olive brines. The total genome sizes varied between 3.77 and 4.039 Mbp. These genome sequences will assist in revealing the genes responsible for both technological and probiotic properties of these strains. ; The Junta de Andalucía supported this study through the P11-AGR-7051 project and the Spanish Government through MICIIN projects AGL2013-48300-R/ALI (OliFilm project) and BFU2016-77297-P. B.C.-D. was the beneficiary of a PIF grant from the Junta de Andalucía. We thank C3UPO for the HPC support. ; Peer Reviewed