Chloride salt mixtures affect Gordal cv. green Spanish-style table olive fermentation

This work studies the effects of different sodium (in the range of 4-10%), potassium (0-4%) and calcium (0-6%) chloride salt mixtures on the fermentation profile of Gordal olives processed according to the Spanish style. For this purpose, response surface methodology based on a simplex centroid mixture design with constrain (sum of salt percentages = 10%) was used. All treatments reached appropriate titratable acidity levels, but this parameter could not be related to the initial chloride salt concentration. The presence of CaCl₂ led to lower initial and after-fermentation pHs, delayed sugar diffusion into the brine, its maximum concentration and titratable acidity formation. CaCl₂ also delayed Enterobacteriaceae and yeast sprang, decreasing their overall growth. This chloride salt also showed a tendency to reduce overall lactic acid bacteria growth. KCl had a similar behaviour to NaCl but, in general, increased overall microbial growth. Thus, a partial substitution of NaCl in Spanish-style green olives with KCl and CaCl₂ does not substantially modify the fermentation profile but does produce some changes, which, when properly managed, could help to improve product processing.     

Biofilm formation on abiotic and biotic surfaces during Spanish style green table olive fermentation

In this work, the establishment of polymicrobial communities on the surfaces which come into contact with the brine during Spanish style Gordal cv. green olive fermentation when subjected to spontaneous or controlled processes (inoculated with Lactobacillus pentosus LPCO10 or 128/2) was studied. Scanning electron microscopy showed that L. pentosus and yeast populations were able to form mixed biofilms throughout the fermentation process on both abiotic (glass slide) and biotic (olive skin) surfaces. The biofilm architectures in both supports were completely different: on the glass slides only aggregates of L. pentosus and yeasts without any polymeric matrix surrounding them were found while on the skin of the fruits, true mature biofilms were observed. During fermentation, the lactic acid bacteria (LAB) population on the olives remained similar while that of yeasts increased progressively to reach similar levels at the end of the process (8-9 log CFU/cm(2)). Molecular analysis showed that different populations of L. pentosus and yeasts were the only microbial members of the biofilm formed during fermentation, regardless of inoculation. Hence, the green olive surface provides an appropriate environmental condition for the suitable development and formation of complex biofilms during controlled or natural table olive processing.     

Ozonation process for the regeneration and recycling of Spanish green table olive fermentation brines

Treatment with ozone broke down the most characteristic polyphenols present in the fermentation brines of green table olives. The ozone required for their complete elimination was 15 and 7 mg/L in acidic solutions (pH 4.0) and alkaline conditions (pH 10.0), respectively. The usual microbial population of brines, lactic acid bacteria and yeasts, was simultaneously eliminated in parallel but needed higher ozone levels in acidic conditions. The ozone treated brines were then filtered through a 0.45 μm pore size and diluted (1:1) and re-used as cover brine for plain and pitted olive packing. After 3 months equilibrium, the cover brines from regenerated brines had slighter poorer colour and slightly higher pH. Fruits packed with diluted (1:1) ozonated brines in alkaline conditions had higher firmness and similar organoleptic characteristics to those using fresh brine. The partial regeneration and reuse of fermentation brines may be a cheap and technologically simple alternative for reducing the environmental problems derived from the high polluting charge of green olive fermentation brines.     

Establishment of conditions for green table olive fermentation at low temperature

Four Lactobacillus plantarum strains were isolated from table olive cold fermentation brines. Their specific growth rate and acidification in MRS broth and in green table olive brines were studied by means of a mixed 2 pH (4.5 and 5.0) x 3 salt (3, 4 and 5%, w/v, NaCl) x 3 incubation temperature (9, 12 and 15 degrees C) levels factorial design. In MRS broth, the greatest effect (linear) on acidification was due to temperature. In brine, the effects were considerably less, pH (linear) being the most important for specific growth rate, and temperature (linear) for acidification. In both media, an initial pH of 5.0 led to good acidification at 12-15 degrees C. The effectiveness of the conditions found (initial pH of 5.0; 3%,w/v, NaCl; and incubation at 12 degrees C) was confirmed in simulated green olive fermentations with three of the strains, which proved especially robust. Behaviour in terms of growth and acidification rates was similar for these strains, and comparable to that observed in traditional processes, although mannitol and sucrose were not metabolised and fructose was only partially used. This leads to the possibility of obtaining normal fermentation processes of table olives in cold regions when appropriate initial conditions and starter cultures are used.     

Reuse of ozonated alkaline solutions as fermentation brines in Spanish green table olives

ABSTRACT:  The water used to reduce the excess NaOH after the immersion of green olives in lye becomes a heavily polluted, alkaline wastewater (AW) once it has come into contact with the fruits. Its treatment with ozonated air for 72 h (TAW) destroyed all polyphenols in the solution. A comparison of the microbial, physicochemical, and organoleptic characteristics of olives processed in the traditional way (W) and those reusing AW or TAW was made. The reuse of TAW or diluted TAW (TAW + W) as fermentation brines led to a shorter lag phase, higher maximum specific growth of lactobacillus, and higher lactic acid accumulation (in TAW) than the traditional process; differences among other physicochemical characteristics were not relevant, except that reused brines (AW) always had higher polyphenol contents. The organoleptic panel test did not detect significant differences among treatments in acidic taste, firmness, pit detachment, or olive color. Overall quality was fairly similar for olives from the traditional process and those from the reused TAW. Direct (or diluted) reuse of AW was also possible but produced a more bitter tasting olive.     

Effect of kaolin and copper based products and of starter cultures on green table olive fermentation

In the present study table olives treated in field with kaolin and copper based products against “olive-fruit fly” were fermented using two selected strains of lactic acid bacteria (LAB). The fermentation process was monitored up to 260 days from brining through physico-chemical, microbiological and sensorial analyses. Results showed a dominance of LAB and yeasts and low level of Enterobacteriaceae counts throughout the whole process both in un-treated and treated samples. When investigating the effect of the single treatments on microbial dynamics, ANOVA results highlighted that copper based products affected significantly the control sample, while the sample inoculated with LAB starters maintained high level throughout the process, guaranteeing the fermentation process. Different behavior was revealed by yeasts population, which was partially influenced by copper treatment at the beginning of the fermentation.The polyphasic approach used in the present study, which combined sensory evaluation to microbial counts and physico-chemical characteristics, let to the conclusion on the importance of starter cultures in fermentation of table olives especially those treated with “non-conventional” pesticide, which could be used to prevent olive fly damage.     

Fermentation profiles of Manzanilla-Aloreña cracked green table olives in different chloride salt mixtures

NaCl plays an important role in table olive processing affecting the flavour and microbiological stability of the final product. However, consumers demand foods low in sodium, which makes necessary to decrease levels of this mineral in fruits. In this work, the effects of diverse mixtures of NaCl, CaCl₂ and KCl on the fermentation profiles of cracked directly brined Manzanilla-Aloreña olives, were studied by means of response surface methodology based in a simplex lattice mixture design with constrains. All salt combinations led to lactic acid processes. The growth of Enterobacteriaceae populations was always limited and partially inhibited by the presence of CaCl₂. Only time to reach half maximum populations and decline rates of yeasts, which were higher as concentrations of NaCl or KCl increased, were affected, and correspondingly modelled, as a function of salt mixtures. However, lactic acid bacteria growth parameters could not be related to initial environmental conditions. They had a longer lag phase, slower growth and higher population levels than yeasts. Overall, the presence of CaCl₂ led to a slower Enterobacteriaceae and lactic acid bacteria growth than the traditional NaCl brine but to higher yeast activity. The presence of CaCl₂ in the fermentation brines also led to higher water activity, lower pH and combined acidity as well as a faster acidification while NaCl and KCl had fairly similar behaviours. Apparently, NaCl may be substituted in diverse proportions with KCl or CaCl₂ without substantially disturbing water activity or the usual fermentation profiles while producing olives with lower salt content.     

Mineral content and sensory characteristics of Gordal green table olives fermented in chloride salt mixtures

This work studies the effects of the initial brine concentrations of NaCl, KCl, and CaCl(2) on the mineral content and gustatory and kinaesthetic sensations of fermented green table olives, using a simplex centroid mixture design augmented with interior points. The sodium in the flesh was linearly related to the mixture concentrations while potassium and calcium were linked by quadratic and special cubic models respectively. Acidity, saltiness, hardness, fibrousness, and crunchiness were expressed as linear funtions of the NaCl, KCl, and CaCl(2) initial brine contents but bitterness required quadratic equations. The models can be used to produce table olives with specific mineral contents in the flesh and to predict their corresponding sensory characteristics. PRACTICAL APPLICATION: The study provides the industry with models to estimate the Na, K, and Ca mineral contents in the flesh of fermented Gordal green table olives as well as their sensory characteristics as a function of the NaCl, KCl, and CaCl(2) initial compositions in the brining solution. Therefore, the paper provides tools which are able to support the production of commercial presentations which not only satisfy consumer demand for low Na, but are also K and Ca fortified table olive presentations with specific sensory profiles.     

Evaluating the individual effects of temperature and salt on table olive related microorganisms

Statistical modelling techniques were used in the present study to assess the individual effects of temperature and NaCl concentration on the growth of 10 lactic acid bacteria and 6 yeast strains mostly isolated from different forms of table olive processing and belonging to the species Lactobacillus pentosus, Lactobacillus plantarum, Saccharomyces cerevisiae, Wickerhamomyces anomalus and Candida boidinii. The mathematical models obtained in synthetic laboratory media show that yeasts, except for C. boidinii, were more resistant to a high salt concentration than lactic acid bacteria, with an MIC value ranging from 163.5 (S. cerevisiae) to 166.9 g/L (W. anomalus); while for L. pentosus and L. plantarum this parameter ranged from 110.6 to 117.6 g/L, respectively. With regards to temperature, lactic acid bacteria showed a slight trend towards supporting higher temperature values than yeasts, with the exception of S. cerevisiae. The maximum temperatures for growth of L. pentosus and L. plantarum were 41.9 and 43.0 °C, respectively; while for W. anomalus and C. boidinii they were 38.2 and 36.5 °C. The optimum temperatures for growth were also higher for L. pentosus and L. plantarum (35.5 and 32.9 °C), compared to W. anomalus and C. boidinii (29.3 and 26.9 °C, respectively). Additional experiments carried out in natural olive brines confirmed previous results, showing that high NaCl concentrations clearly favoured yeast growth and that at high temperatures LAB slightly overcame yeasts. Results obtained in this paper could be useful for industry for a better control of both table olive fermentation and packaging.     

Spanish‐style green table olive shelf‐life

Changes in the physicochemical and sensory characteristics of commercial whole, pitted and stuffed with pepper, garlic or anchovy paste Spanish‐style green table olives of the Gordal, Manzanilla and Hojiblanca cultivars were studied during a three‐year period. No olives developed any spoilage during this period, and at the end of it, all presentations were classified as ‘extra’ according to the sensory evaluation methodology issued by the International Olive Council. Throughout the shelf‐life, surface colour index and firmness of the olives degraded in accordance with a first‐order kinetics, while a gradual browning of the cover brines was observed; however, products added with ascorbic acid maintained a stable colour, while those with calcium content showed reduced firmness degradation. No significant changes in most of the sensory characteristics were observed by the panel test during shelf‐life and only slight changes in firmness, and surface colour were noticed in samples with significant differences in these attributes when measured instrumentally.     

Use of selected autochthonous lactic acid bacteria for Spanish-style table olive fermentation

The present work presents a successful attempt to achieve an enhanced and more predictable fermentation process in Spanish-style green olive technology by selection and use of autochthonous starter cultures. During the first phase of this work, two Spanish-like fermentations of green table olives of cultivar (cv) "Nocellara del Belice", coming from irrigated and not irrigated fields, were monitored, in order to highlight the best agricultural conditions for drupe production and to isolate lactic acid bacteria strains with relevant technological properties. Among 88 identified isolates, one Lactobacillus pentosus strain showed remarkable biochemical features and high acidification rate in synthetic brine. In the second phase, the selected strain was used as starter culture in three different trials to establish the best conditions for its use. Microbial counting, as well as starter tracking by M13 RAPD-PCR, reflected the optimal adaptation of the strain to the environment. Spontaneous fermentation needed a 14-day long lag phase to reach the same population as the inoculated trials. Moreover, sensory traits of table olives obtained with adjunct culture showed better characteristics compared to those processed in the other trials, in particular concerning the presence of off-odours.     

Chemical composition changes in four green olive cultivars during spontaneous fermentation

Changes in some physico-chemical characteristics (pH and free acidity) and chemical composition (sugars, total phenolic and flavonoid contents) of four olive cultivars during spontaneous fermentation in brine were investigated. The cultivars were typical of the Moroccan market: “Moroccan Picholine”, “Languedoc Picholine”, “Ascolana” and “Sevillana”. The physico-chemical changes of olives and brines during fermentation process were monitored. A similar pattern of pH was noticed for the “Moroccan Picholine”, “Languedoc Picholine” and “Ascolana” cultivars with a final pH ranging between 4.4 and 4.6. The profile of free acidity measured throughout fermentation period in brines was in agreement with the pH trend. The concentration of sugars, total phenolic and flavonoids contents in olives flesh and brines during fermentation is reported. The loss of flavonoids, sugars and total phenolic contents in the olive flesh by the end of fermentation process was up to 60%, 63% and 79% in “Languedoc Picholine”, “Sevillana” and “Moroccan Picholine”, respectively. The main phenols identified and quantified in the different brines at the end of brining process were hydroxytyrosol, tyrosol, (+)-catechin and quercetine. The highest total phenolic content and antioxidant activity were obtained in Moroccan Picholine brine after 71 days of fermentation.     

Exploring the yeast biodiversity of green table olive industrial fermentations for technological applications

In recent years, there has been an increasing interest in identifying and characterizing the yeast populations associated with diverse types of table olive elaborations because of the many desirable technological properties of these microorganisms. In this work, a total of 199 yeast isolates were directly obtained from industrial green table olive fermentations and genetically identified by means of a RFLP analysis of the 5.8S-ITS region and sequencing of the D1/D2 domains of the 26S rDNA gene. Candida diddensiae, Saccharomyces cerevisiae and Pichia membranifaciens were the most abundant yeast species isolated from directly brined Aloreña olives, while for Gordal and Manzanilla cultivars they were Candida tropicalis, Pichia galeiformis and Wickerhamomyces anomalus. In the case of Gordal and Manzanilla green olives processed according to the Spanish style, the predominant yeasts were Debaryomyces etchellsii, C. tropicalis, P. galeiformis and Kluyveromyces lactis. Biochemical activities of technological interest were then qualitatively determined for isolates belonging to all yeast species. This preliminary screening identified two isolates of W. anomalus with interesting properties, such as a strong β-glucosidase and esterase activity, and a moderate catalase and lipolytic activity, which were also confirmed by quantitative assays. The results obtained in this survey show the potential use that some yeast species could have as starters, alone or in combination with lactic acid bacteria, during olive processing.     

Study of green Sicilian table olive fermentations through microbiological,chemical and sensory analyses

The production of five different green table olive cultivars was studied by a combined strategy consisting of chemical, microbiological and sensory analyses. Cultivable microflora of samples collected during processing was monitored by plate counts on seven synthetic culture media. In all samples Enterobacteriaceae, Pseudomonaceae, staphylococci, lactic acid bacteria and spore-forming bacteria were undetectable. Yeasts and moulds were countable from the day 42 (2 log CFU/ml) till the end of fermentation (6 log CFU/ml). The use of three different approaches for microorganism detection, including a culture-independent methodology, revealed the presence of barely three yeast species during the entire fermentation period: Candida parapsilosis, Pichia guilliermondii and Pichia kluyveri. Biochemical features of technological interest were evaluated for 94 strains in order to investigate their potential role in fermentation of green Sicilian table olives.     

Inhibitors of lactic acid fermentation in Spanish-style green olive brines of the Manzanilla variety

Frequently, a delay or lack of lactic acid fermentation occurs during the processing of Spanish-style green olives, in particular of the Manzanilla variety. Many variables can affect the progress of fermentation such as temperature, nutrients, salt concentration, antimicrobials in brines, and others. In this study, it was demonstrated that an inappropriate alkaline treatment (low NaOH strength and insufficient alkali penetration) allowed for the presence of several antimicrobial compounds in brines, which inhibited the growth of Lactobacillus pentosus. These substances were the dialdehydic form of decarboxymethyl elenolic acid either free or linked to hydroxytyrosol and an isomer of oleoside 11-methyl ester. Olive brines, from olives treated with a NaOH solution of low concentration up to 1/2 the distance to the pit, contained these antimicrobials, and no lactic acid fermentation took place in them. By contrast, a more intense alkaline treatment (2/3 lye depth penetration) gave rise to an abundant growth of lactic acid bacteria without any antimicrobial in brines. Therefore, the precise cause of stuck fermentation in Manzanilla olive brines was demonstrated for the first time and this finding will contribute to better understand the table olive fermentation process.     

Utilization at high pH of starter cultures of lactobacilli for Spanish-style green olive fermentation

Inoculation at alkaline pH (above 9) of lye-treated green olives with starter cultures of Lactobacillus pentosus CECT 5138 was studied. Despite an initial loss of viability in the order of 1-2 log cycles on average, depending mainly on time of application, cultures grew and initiated an accelerated fermentation process. Inoculation reduced the population of Enterobacteriaceae, and thereby potential spoilage, and produced a quicker acidification of brines and decrease of pH, when compared with control uninoculated batches. Results obtained throughout three consecutive seasons demonstrated that utilization at high pH of starter cultures of lactobacilli is feasible, provided that the inoculum size takes into account the initial low survival.     

碘盐及发酵温度对低盐固态酱油质量的影响

实验室条件下考察了碘盐及发酵温度对低盐固态法酿造酱油的影响。试验分析表明微量碘对酱油的色泽有一定影响,使用碘盐的酱油色率略高于非碘盐的酱油,红色指数相近。而发酵温度按第1～7d,41～43℃;第8～18d,44～46℃;第19～20d,47～48℃模式控制,低盐固态酱油的色率及红色指数指标较好,氨态氮的生成率最高。     

Effects of pasteurisation and storage on quality characteristics of table olives preserved in olive oil

Two types of products obtained from the processing of Olea europaea cv ‘Taggiasca’ were monitored: pitted olives and olive paste preserved in monovarietal extra‐virgin olive oil of the same variety. The aim of this study was to follow the trends in the main physico‐chemical and sensory parameters related to the quality of these table olives and their covering oils, for simulation of the shelf‐life conditions. From all of the analyses carried out, it can be concluded that the optimum time of storage of these two products packaged in glass jars at room temperature (between 18 and 25 °C) and under artificial light and away from heat sources is approximately 9 months. After 9 months of storage, the covering oils were greatly affected by the contact with the broken fruit, which was accompanied by reductions in their antioxidant content due to the thermal treatment. The oxidative effects seen through physico‐chemical analyses are in agreement with the organoleptic analysis.