A New Natural Processing System Based on Slight Carbon Dioxide Pressure for Producing Black Table Olives with Low Salt Content.

Naturally fermented black table olives are usually processed in brine with low pH and high NaCl content. Because salt is responsible for several cardiovascular problems, methods are needed to decrease the salt (NaCl) content in olive pulp. This study investigated a new natural processing system wherein microorganism growth is inhibited by slight pressure of CO2 (spCO2), in addition to low pH and NaCl, in brine with decreased salt content. The fermentation performed under spCO2 with a low-salt brine with 6% (w v-1) NaCl and 0.5% (w v-1) citric acid, unlike the traditional system, inhibited the growth of bacteria and fungi and decreased the concentration of yeasts. Processing tests with spCO2 in the presence of different salt and citric acid concentrations indicated a slight decrease in yeasts in brines containing 6% (w v-1) NaCl and 0.6% (w v-1) citric acid but not after inoculation of the same brines with Saccharomyces cerevisiae. In contrast, in the presence of 11% (w v-1) NaCl and 0.3% or 0.6% (w v-1) citric acid, the inhibitory effect of brines was greater compared to those with low-salt and it was also confirmed in the same brines inoculated with S. cerevisiae.

Use of Slightly Pressurized Carbon Dioxide to Enhance the Antimicrobial Properties of Brines in Naturally Processed Black Table Olives.

Naturally fermented black table olives are processed at low pH in the presence of high sodium chloride concentrations ranging from 8 to 12% (w v-1). Reducing the salt content of brine has become an urgent issue as it is responsible for several health and environmental problems. The study aim was to evaluate slightly pressurized CO2 (spCO2) as a third barrier to microbial growth in naturally processed black table olives with low pH and a reduced NaCl concentration. Based on the assessments performed on a pilot plant scale, an spCO2 of 1 bar completely inhibited the growth of the bacteria and molds in the presence of reduced saline concentrations. Furthermore, the amount of yeast decreased in the brine as a function of the NaCl content. Laboratory tests performed under spCO2 conditions using a single yeast species from the same habitat confirmed the high sensitivity of some oxidizing yeasts and indicated that the fermenting yeast, Saccharomyces cerevisiae, is the most tolerant species. Overall, in the brine of naturally processed olives with a low pH between 4 and 4.2, the antimicrobial properties observed with the high concentrations of NaCl can be achieved with a lower salt dose of 5% (w v-1) when combined with spCO2.

Microbiological and Enzymatic Activity Modulates the Bitter Taste Reduction in Decanted Coratina Olive Oil.

Coratina monocultivar extra virgin olive oil (EVOO) is known for its level of bitterness, which, if too high, can cause consumer acceptance problems. The aim of this study was to modulate the bitter taste of freshly produced olive oil through endogenous enzymatic activity and microbiota during the decantation phase. The opalescent appearance of the newly produced EVOO was substantially reduced during the first three months of decantation due to the deposition of more than 90% of suspended material, consisting of vegetation water and suspended solid particles. The high content of biophenols and the reduction in water concentration in the oil samples negatively affected the survival of yeasts, which were absent in the oil samples at the end of the third month of decantation. The oleuropeinolytic activity was very intense during the first month of decantation, whereas the reduction in the bitter taste associated with the aglycons was consistent only in the second and third months of decantation. At the end of decantation, the sensory notes of bitterness in the Coratina EVOO were reduced by 33%, lowering the position on the value scale without altering the other qualitative parameters whose values fell within the limits of the commercial EVOO class.

Role of yeasts in the qualitative structuring of extra virgin olive oil.

This review sought to describe the role played by some components of the microbiota of extra virgin olive oil (EVOO), particularly yeasts, in structuring the physicochemical and sensorial quality of freshly produced olive oil. Yeasts can survive during the entire storage period of the product. To date, approximately 25 yeast species isolated from oil produced in more than six countries have been identified, eight of which are classified as new species. Some yeast species improve the health qualities of oil, whereas many others improve the chemical composition and sensory characteristics based on ß-glucosidase and esterase enzymes, which are involved in the hydrolysis of the bitter glucoside known as oleuropein. However, some species, which are typically favoured by the high water content in the oily matrix, such as lipase-producing yeasts, can worsen the initial chemical characteristics of EVOO oil during storage. Some physical treatments that are compatible with the EVOO production specification affect the biotic component of the oil by reducing the concentration of yeasts. The possibility of minimizing the invasive action on the biotic component of the oil by appropriately selecting the physical treatment for each oil is discussed.

Effects of the Filtration on the Biotic Fraction of Extra Virgin Olive Oil.

Filtration is a widely used process in the production of extra virgin olive oil. We studied the influence of filtration performed with cotton filters and cellulose filter press on the biotic components of the oily mass containing probiotic traits in two freshly produced monocultivar extra virgin olive oils. The concentration of bacteria was reduced from 100% to 28%, while that of fungi was reduced from 100% to 44% after filtration, according to the filtration system and the initial contamination of the original monocultivar extra virgin olive oil. Compared with the control, the yeast content in the oil samples filtered with cotton filters was reduced from 37% to 11% depending on the cultivar. In the oil filtered with cellulose filter press, the yeast content reduced from 42% to 16%. The viable yeast that passed through the oily mass during the filtration process with cellulose filter press, unlike all the other samples, were unable to survive in the oil after a month of storage. The possible health benefits of compounds from both the biotic and abiotic fraction of the oil, compared to the control, were significantly low when filtered with the cellulose filter press.

Use of Air-Protected Headspace to Prevent Yeast Film Formation on the Brine of Leccino and Taggiasca Black Table Olives Processed in Industrial-Scale Plastic Barrels.

The formation of yeast film on the brine of black table olives during fermentation in plastic barrels on an industrial-scale could be critical for the quality of the product. In order to prevent the formation of yeast film on the brine surface, a structural modified industrial barrel, which excludes oxygen from the headspace, was tested. Tests carried out during two years indicated that the yeast film contamination reached the maximum values at eight months of fermentation, equal to 19% and 24% respectively, for the Taggiasca and Leccino olives, processed in unmodified industrial plastic barrels. No yeast films formed on brines from the same varieties of olives processed in the modified plastic barrels. The brines of both varieties of olives processed in the industrial barrels displayed three dominant yeast species, while five species were detected in the brines from the modified barrels. Wickerhamomyces anomalus and Pichia manshurica were the main producers of yeast films. However, P. manshurica unlike the other yeasts, has shown also a biotype unable to produce films on the brine of the olives. The brines of Leccino and Taggiasca processed in the modified barrels, compared to the control, showed a higher titratable acidity and a higher concentration of CO2 useful to prevent the yeast film formation.

Virgin Olive Oil Quality Is Affected by the Microbiota that Comprise the Biotic Fraction of the Oil.

This review summarizes the current knowledge on the effects of oil-borne yeasts on the physicochemical, sensorial, and health-related characteristics of virgin olive oil (VOO) during storage. Bacteria, yeasts, and molds constitute the biotic fraction of freshly produced VOO. During storage, the bacteria and molds often die after a short period, while the yeasts survive and condition the quality of VOO. To date, approximately twenty-four yeast species have been isolated from different types of olive oil and its by-products, and seven of these species have been identified as new species. The activity of some yeasts of the biotic fraction of olive oil improves the sensorial characteristics of VOO. Some yeasts can also worsen the quality of the product by allowing the appearance of defects, oxidation of polar phenols, and triacylglycerol hydrolysis. Some yeast species of VOO show in vitro beneficial health effects, such as probiotic and antioxidant activities.

Differential Microbial Composition of Monovarietal and Blended Extra Virgin Olive Oils Determines Oil Quality During Storage.

Extra virgin olive oil (EVOO) contains a biotic fraction, which is characterized by various microorganisms, including yeasts. The colonization of microorganisms in the freshly produced EVOO is determined by the physicochemical characteristics of the product. The production of blended EVOO with balanced taste, which is obtained by blending several monovarietal EVOOs, modifies the original microbiota of each oil due to the differential physico-chemical characteristics of the blended oil. This study aimed to evaluate the effect of microbial composition on the stability of the quality indices of the monovarietal and blended EVOOs derived from Leccino, Peranzana, Coratina, and Ravece olive varieties after six months of storage. The yeasts survived only in the monovarietal EVOOs during six months of storage. Barnettozyma californica, Candida adriatica, Candida diddensiae, and Yamadazyma terventina were the predominant yeast species, whose abundance varied in the four monovarietal EVOOs. However, the number of yeasts markedly decreased during the first three months of storage in all blended EVOOs. Thus, all blended EVOOs were more stable than the monovarietal EVOOs as the abundance and activity of microorganisms were limited during storage.

Survival of Coliform Bacteria in Virgin Olive Oil.

Coliform bacteria consist of both nonpathogen commensal and human opportunistic pathogen species isolated from different habitats like animals, man, vegetables, and water. Olives normally carry natural nonpathogenic epiphytic bacteria, but during growth, harvest, and processing, one of the final products, represented by virgin olive oil, can be contaminated with coliform. Present study showed that coliform bacteria can survive and reproduce in virgin olive oil containing low level of phenolic compounds. The laboratory inoculation trials demonstrated that when the bacterium Escherichia coli, isolated from the olives carposphere, was transferred in olive oil containing high polar phenols content, equal to 372 mg caffeic acid equivalent per kg, the survival was completely inhibited after 15 days of storage. On the contrary, the bacterium reproduced quickly when it was inoculated in virgin olive oil samples containing lower concentration of polar phenols. The SDS-PAGE analysis of the E. coli proteins showed different electrophoretic patterns when the bacterium was inoculated in the virgin olive oil with high phenolic compounds content, confirming the strong interaction between the olive oil phenols content and the bacterial wall proteins. The SEM ultrastructural observations confirmed the presence of a more higher number of damaged microbial cells in virgin olive oil rich of polar phenols. This finding needs further studies since, in an era of antibiotic resistance, the development of new strategies to fight unwanted food bacteria is promising way for the future.

Virgin olive oil yeasts: A review.

This review summarizes current knowledge on virgin olive oil yeasts. Newly produced olive oil contains solid particles and micro drops of vegetation water in which yeasts reproduce to become the typical microbiota of olive oil. To date, about seventeen yeast species have been isolated from different types of olive oils and their by-products, of which six species have been identified as new species. Certain yeast species contribute greatly to improving the sensorial characteristics of the newly produced olive oil, whereas other species are considered harmful as they can damage the oil quality through the production of unpleasant flavors and triacylglycerol hydrolysis. Studies carried out in certain yeast strains have demonstrated the presence of defects in olive oil treated with Candida adriatica, Nakazawaea wickerhamii and Candida diddensiae specific strains, while other olive oil samples treated with other Candida diddensiae strains were defect-free after four months of storage and categorized as extra virgin. A new acetic acid producing yeast species, namely, Brettanomyces acidodurans sp. nov., which was recently isolated from olive oil, could be implicated in the wine-vinegary defect of the product. Other aspects related to the activity of the lipase-producing yeasts and the survival of the yeast species in the flavored olive oils are also discussed.

Yamadazyma terventina sp. nov., a yeast species of the Yamadazyma clade from Italian olive oils.

During an investigation of olive oil microbiota, three yeast strains were found to be divergent from currently classified yeast species according to the sequences of the D1/D2 domain of the gene encoding the rRNA large subunit (LSU) and the internal transcribed spacer region including the gene for 5.8S rRNA. Phylogenetic analysis revealed that these strains, designated CBS 12509, CBS 12510(T) and CBS 12511, represent a novel anascosporogenous species described herein as Yamadazyma terventina sp. nov; the type strain is DAPES 1924(T) (= CBS 12510(T) = NCAIM Y.02028(T)). This novel species was placed in the Yamadazyma clade, with Yamadazyma scolyti, Candida conglobata and Candida aaseri as closest relatives. Y. terventina differs from the above-mentioned species in the ability to strongly assimilate dl-lactate and weakly assimilate ethanol.

Candida adriatica sp. nov. and Candida molendinolei sp. nov., two yeast species isolated from olive oil and its by-products.

Thirteen strains isolated from virgin olive oil or its by-products in several Mediterranean countries were found to be phenotypically and genetically divergent from currently recognized yeast species. Sequence analysis of the large subunit (LSU) rDNA D1/D2 domain and internal transcribed spacer regions/5.8S rDNA revealed that the strains represented two novel species described as Candida adriatica sp. nov. (type strain ZIM 2334(T) = CBS 12504(T) = NCAIM Y.02001(T)) and Candida molendinolei sp. nov. (type strain DBVPG 5508(T) = CBS 12508(T) = NCAIM Y.02000(T)). Phylogenetic analysis based on concatenated sequences of the small subunit rRNA gene, the D1/D2 region of the LSU rDNA and the translation elongation factor-1α gene suggested that C. adriatica sp. nov. and C. molendinolei sp. nov. should be placed within the Lindnera and Nakazawaea clades, respectively.

Evaluation of Polymyxa betae Keskin Contaminated by Beet Necrotic Yellow Vein Virus in Soil.

The fungus Polymyxa betae Keskin belongs to the family Plasmodiophoraceae and lives in the soil as an obligatory parasite of the roots of the Chenopodiaceae. When contaminated by beet necrotic yellow vein virus, this viruliferous fungus causes a serious disease of sugar beet known as rhizomania, whereas the infection by the fungus alone (aviruliferous fungus) causes only slight damage to the plant with little economic consequence. The manifestation of rhizomania in sugar beet is directly related to the concentration of infecting units of viruliferous P. betae present in the soil. (One infecting unit is a group of one or more sporosori that liberate zoospores capable of visibly infecting a plant.) By using current methods of analysis, it is possible to estimate the total quantity of P. betae present in the soil, but one cannot distinguish quantitatively the infecting units of aviruliferous from viruliferous P. betae. A new method has been developed based on the technique of the most probable number and enzyme-linked immunosorbent assay to estimate the concentration of infecting units of viruliferous P. betae in soil. The method is suitable for the routine analysis of numerous soil samples and allows one to estimate the concentration of viable forms of the fungus P. betae, whether or not contaminated by beet necrotic yellow vein virus, present in a soil affected by rhizomania or presumed healthy. The analyses performed with this method are economical and use a reagent kit and equipment in wide use.