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.     

A study on the implications of NaCl reduction in the fermentation profile of Conservolea natural black olives

This study examined the impact of different mixtures of NaCl, KCl, and CaCl₂ on the fermentation profiles of Conservolea natural black olives. Five different combinations of chloride salts were investigated, namely (i) 8% NaCl (control treatment), (ii) 4% NaCl and 4% KCl, (iii) 4% NaCl and 4% CaCl₂, (iv) 4% KCl and 4% CaCl₂, and (v) 2.6% NaCl-2.6% KCl-2.6% CaCl₂. The changes in the microbial association (lactic acid bacteria, yeasts, Enterobacteriaceae), pH, titratable acidity, organic acids, volatile compounds, and mineral content in olive flesh were analyzed. Results demonstrated that all salt combinations led to vigorous lactic acid processes based on the obtained values of pH (3.9-4.2) and titratable acidity (0.70-0.86 g lactic acid per 100 ml brine). Organoleptic evaluation was a critical factor in the acceptability of the final product. Increasing concentrations of CaCl₂ or a combination of KCl and CaCl₂ rendered the product bitter with low acceptability by the taste panel. Only one combination of chloride salts (4% NaCl and 4% KCl) could finally produce olives with lower sodium content and good organoleptic attributes. The results of this study could be employed by the Greek table olive industry in an attempt to produce natural black olives with less sodium without affecting the traditional taste of fermented olives in order to meet consumers’ demand for low sodium dietary intake.     

Calcium Chloride and Potassium Sorbate Reduce Sodium Chloride used during Natural Cucumber Fermentation and Storage

Cucumber fermentation characteristics and pickle quality were evaluated in brines containing equilibrium concentrations of 0-0.4% CaCl₂, 0-0.4% potassium sorbate and 0-10% NaCl. Changes in brine pH and acidity, cucumber texture and color, coliforms, lactic acid and total bacteria, yeasts and molds were followed over time. Results indicated that cucumber spoilage would eventually take place if NaCl or potassium sorbate were not present in the brine. The presence of CaCl₂ helped maintain cucumber firmness. A synergistic action between NaCl, CaCl₂ and potassium sorbate was seen, which allowed good quality pickles to be produced when moderate amounts of all three components were present in the brine (5% NaCl, 0.2% CaCl₂, 0.2% potassium sorbate).     

Aerobic industrial processing of Empeltre cv. natural black olives and product characterisation

The fermentation of natural black olives is currently carried out under anaerobic conditions. This work investigates the aerobic fermentation of this product in tanks containing 16 000 kg of Empeltre olives during two seasons. The microorganisms in brines were yeasts in all cases, whereas lactic acid bacteria growth was only observed in some tanks. Consequently, the pH during fermentation was in most cases higher than recommended 4.3 units. However, the Empeltre olive brines displayed bactericidal effect against Salmonella enterica, Sthapylococcus aureus, Escherichia coli and Listeria monocytogenes, with a 5 log population drop in one or 24 h depending on the brine age so that these olive brines are a harsh environment for the growth of these pathogens. It was found HyEDA as the major phenolic compound in both brine and olive pulp at the beginning of fermentation that slowly hydrolysed into hydroxytyrosol that reached a concentration up to 1500 mg kg⁻¹.     

Effect of different packing treatments on the microbiological and physicochemical characteristics of untreated green olives of the Conservolea cultivar

Microbiological, physicochemical and organoleptic changes were studied in non‐pasteurised samples of untreated green olives cv Conservolea stored at 20 °C for 180 days under different preservation treatments (acidified brine and modified atmospheres). The microbial flora consisted of lactic acid bacteria and yeasts, while no enterobacteria and pseudomonads were enumerated in any package. Although no thermal treatment was applied to the packages, no sediment formation was observed due to the growth of propionic bacteria. Olives packed in brine supplemented with 0.3% lactic and 0.3% citric acids presented the highest acidity, the lowest brine and flesh pH and the lowest dark brine colour as measured by the parameter A₄₄₀?A₇₀₀. Olives packed in modified atmospheres (vacuum, 40% CO₂/30% O₂/30% N₂ and air) presented a progressive loss of fruit firmness and colour with time. At the end of the storage period, olives packed in aerobic conditions and 40% CO₂/30% O₂/30% N₂ presented the lowest firmness and skin colour and were unacceptable to the panellists. The best quality characteristics were maintained in vacuum‐packed olives, as indicated by the higher organoleptic scores. Firmness degradation followed first‐order kinetics (r² > 0.980) and the expected shelf‐life was 23, 15 and 9 months for olives packed in vacuum, 40% CO₂/30% O₂/30% N₂ and air respectively. Copyright © 2004 Society of Chemical Industry     

Microbiological and physicochemical changes of naturally black olives fermented at different temperatures and NaCl levels in the brines

The effect of different temperatures (25°C, 18°C and ambient temperature) and NaCl levels in brines (4%, 6% and 8%) on the microbiological and physicochemical characteristics of naturally black olives of Conservolea variety was studied for up to 190 days. Fermentation was carried out according to the traditional anaerobic method. The initial microflora consisted of Gram-negative bacteria, lactic acid bacteria and yeasts. Inhibition of Gram-negative bacteria was evident in all fermentations. The prevailing micro-organisms were lactic acid bacteria and yeasts, the association of which was dependent on the conditions of fermentation. At 25°C and 18°C in brines containing 4% and 6% NaCl, the growth of lactic acid bacteria was favoured resulting in a lactic acid fermentation, as indicated by the high free acidity levels and low pH values in the brines. On the contrary, 8% NaCl concentration affected the growth of lactic acid bacteria and enhanced the activity of fermentative yeasts, producing a final product with lower free acidity and higher pH value. At ambient temperature, the counts of lactic acid bacteria followed the fluctuation of temperature regardless of salt concentration, while yeasts did not seem to be affected. The lactic acid bacteria identified belonged to the species Lactobacillus mesenteroides, Lactobacillus brevis, Lactobacillus plantarum and Lactobacillus pentosus. The best conditions for fermentation were at 25°C and 6% NaCl, developing free acidity of 142 mM (1·28% w/v) lactic acid and pH value of 3·8. After 5 months of brining, olives fermented at 25°C were judged by panelists as being debittered and ready to eat. No off-odour development was detected in any case due to anomalous fermentation. The HPLC analysis revealed that citric, malic, tartaric, succinic, lactic and acetic were among the end products of fermentation.     

New genotypes of table olives: profile of bioactive compounds

New table olive genotypes (48) coming from a cross‐breeding programme were evaluated. Most of the fruit traits covered a wide range of variability on the set of genotypes, fruit weight (1.1–9.7 g), pulp‐to‐pit ratio (1.7–10.0), fruit shape (1.0–1.6) and oil content (1.3–15.2%). This is the first time that healthy compounds such as triterpenic acids and phenolic compounds have also been evaluated in olive progenies. Genotypes were stored for 2 months in sterilised brine (5% NaCl and 0.5% acetic acid). A high amount of maslinic (685.0–1394.2 mg kg^?1 olive flesh) and oleanolic acids (275.3–817.9 mg kg^?1 olive flesh) was found in the flesh of olives stored. The main oleosidic and phenolic compounds evaluated in brines were hydroxytyrosol (1.9–8.4 mmol L^?1), hydroxytyrosol glucosides (0.4–19.8 mmol L^?1), oleuropein (0.0–4.7 mmol L^?1) and the antimicrobial compounds, dialdehydic form of decarboxymethyl elenolic acid linked to hydroxytyrosol (0.0–3.4 mmol L^?1) and decarboxymethyl elenolic acid (0.0–1.7 mmol L^?1), the latter two being observed in only ten genotypes. The wide range of variation observed for most compounds indicates that the contents of these healthy compounds may be used as selection criteria in table olive breeding programmes.     

Evaluation of a single and combined inoculation of a Lactobacillus pentosus starter for processing cv. Arbequina natural green olives

The production of Arbequina naturally green olives is a traditional and spontaneous process in which lactic acid bacteria (LAB) and yeasts are present. To better control the fermentation of olives, strains of LAB and yeasts that had been isolated from brines were used in this study.     

Effect of pretreatments and drying temperatures on sweet potato flour

Effect of pretreatments with 1 w/v% sodium hydrogen sulphite (NaHSO₃) and 1 w/v% calcium chloride (CaCl₂) and drying temperatures (55, 60 and 65 °C) on sweet potato flour were investigated. Flour treated with CaCl₂ had higher amounts of ascorbic acid and β‐carotene (10.61–12.54 and 3.26–3.46 mg 100 g^?1 wet basis, respectively) than that treated with NaHSO₃ (9.47–11.47 and 3.05–3.43 mg 100 g^?1 wet basis, respectively). Total phenolic content and water absorption index (wet basis) were highest at 65 °C when treated with NaHSO₃ (10.44 mg 100 g^?1 and 2.49 g g^?1 respectively) and CaCl₂ (9.52 mg 100 g^?1 and 2.85 g g^?1 respectively). Swelling capacity (wet basis) was highest at 60 °C when treated with CaCl₂ (2.96 g g^?1) whereas when treated with NaHSO₃ (2.85 g g^?1) it was highest at 55 °C. Freeze‐dried samples treated with NaHSO₃ had higher lightness and total phenolic content while CaCl₂‐treated samples had higher β‐carotene and ascorbic acid. The results showed that good quality flour could be produced after soaking in CaCl₂ and dried at 65 °C.     

Catalase, Lipoxygenase, and Peroxidase Activities in Cucumber Pickles as Affected by Fermentation, Processing, and Calcium Chloride

Catalase, lipoxygenase, and peroxidase activities were determined in tissues and brines of pickling cucumbers during fermentation, storage, and after processing. Lipoxygenase and catalase were inactivated within three days of exposure to fermentation brines. Peroxidase activity was relatively unaffected by the first three days of brining, but it declined during fermentation, storage, and after processing. CaCl₂ reduced the loss of peroxidase activity in tissue and brine during fermentation and storage. Alum in post-desalting brine reduced peroxidase activity in processed pickles although CaCl₂ ameliorated its efficacy. No activity of catalase, lipoxygenase or peroxidase was detected in cucumber juice extracts fermented by four different lactic acid bacteria.     

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.     

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.     

Improvement of the storage process for cracked table olives

The aim of this work was to improve the storage method for cracked table olives. First, a study was carried out of the washing phase in 5% (wt/vol) NaCl brine with ascorbic acid and sodium metabisulphite added to it. Its use removed half the proportion of the initial sugar content in the flesh and markedly decreased the content of polyphenols in the fruits, confirming the effectiveness of this treatment. Washing with 5% NaCl and sodium metabisulphite was selected and applied in storage phase which follows. Storage of the washed cracked olives produced an additional decrease in sugars and polyphenols in the flesh, although the product still retained a fairly high concentration of residual sugars at the end of the storage period. The growth of lactic acid bacteria in this phase was prevented by the high NaCl level (10–13% at the equilibrium) but yeast growth, followed by a decline phase, was found in all storage systems studied. The use of a CO₂ atmosphere led to a partial inactivation of yeasts for about 300 h and showed a protective effect on polyphenols. Application of ozone had a favourable effect on initial solubilisation of sugars and polyphenols but caused a negative effect on fresh appearance, colour retention and polyphenol degradation. CaCl₂ + MgCl₂ treatment led to a significantly faster decline phase of yeasts. A tentative improved process for cracked olive storage may consist of a previous washing of olives for 24 h with a 5% NaCl solution (optionally added with 0.1% sodium metabisulphite), and a subsequent brining in a 15% NaCl solution with 0.25% CaCl₂ + 0.20% MgCl₂ or in a 15% NaCl brine under CO₂ atmosphere.     

Efficacy of natamycin to control fungal growth in natural black olive fermentation

The effect of natamycin as a fungal control agent in natural black olive fermentation according to the traditional anaerobic system was studied. Black Conservolea olives were subjected to spontaneous fermentation in 8% (w/v) NaCl brine solution (control treatment) or brine supplemented with 0.01% (w/v) natamycin (active compound) for an overall period of 60 days. The changes in the microbial association (lactic acid bacteria, yeasts, Enterobacteriaceae), pH, titratable acidity, organic acids, and volatile compounds were monitored throughout fermentation. The initial microbiota consisted of lactic acid bacteria, yeasts, and Enterobacteriaceae. The addition of natamycin in the brine inhibited the growth of yeasts, without affecting the population dynamics of bacteria, resulting in a more vigorous fermentation with higher titratable acidity compared to spontaneous control process. Moreover, the presence of natamycin inhibited mould spoilage caused by the development of fungal mycelium on the surface of the brine during the traditional anaerobic fermentation system employed widely in Greece in natural black olive processing. Natamycin could be an important component of a processing system to control fungal growth in natural black olive fermentation and at the same time it has the potential to enhance the process by favouring the growth of the indigenous population of lactic acid bacteria against other competing microorganisms.     

Effect of brine replacement and salt concentration on the fermentation of naturally black olives

This study was undertaken to modify the traditional fermentation of naturally black olives in order to obtain higher quality black table olives. After holding olives in tap water or low salt brine prior to fermentation, acidification of the cover brine and fermentation at two different salt concentrations (6 and 14 g/ 100 ml NaCl) were applied to Gemlik variety olives in three batches. Fermentations were monitored using physical, chemical and microbiological analyses and effects of the above changes to fermentation were investigated. Yeasts were the predominant microorganisms in all of the batches. Growth enhancement of lactic acid bacteria by brine replacement occurred in the fermentation with 6 g/ 100 ml NaCl cover brine but not with 14 g/100 ml NaCl brine. The highest level of titratable acidity (0.59 g/100 ml) formed during fermentation with 6 g/100 ml NaCl brine followed by fermentation with 6 g/100 ml NaCl brine (0.43 g/100 ml) where the brine was replaced after 48 days. The lowest level of titratable acidity (0.36 g/100 ml) was formed using a high salt brine. Reducing sugars were consumed effectively by microorganisms in all the batches and residual levels in the brines were 0.05–0.1 g/100 g ml. NaCl contents of the final products were 5.20 g/ 100 g (olives with high salt brine) and 3.27–3.58 g/100 g (olives with low salt brine). Moisture contents of the processed olives were in the range 49.58–50.79 g/ 100 g. The water activity (a_w) value of olives fermented with the high salt brine was found to be 0.947 while a_w values of olives fermented with the low salt brine were 0.962–0.964. Protein content of the olives ranged between 1.76–1.95 g/100 g and the ash contents were found to be 2.41–2.81 g/100 g in olives with low salt brine and 4.70 g/100 g in olives with high salt brine. According to sensory evaluations of the final products, no differences between the treatments were found (P > 0.05).     

Simultaneous lactic acid and xylitol production from vine trimming wastes

Hemicellulosic hydrolyzates from vineshoot trimmings obtained by dilute sulfuric acid hydrolysis were evaluated for xylitol production by Debaryomyces hansenii NRRL Y‐7426. Bioconversion was not efficient, however, since a mixture of products (mainly ethanol) was achieved. Taking into account that hexoses (such as glucose or mannose) can inhibit xylose metabolism by repression and inactivation of the xylose transport system or catabolic enzymes and that these hemicellulosic hydrolyzates are characterized by a high glucose concentration, a novel technology was developed, sequentially transforming glucose into lactic acid by Lactobacillus rhamnosus followed by fermentation of xylose into xylitol by Debaryomyces hansenii after L. rhamnosus removal by microfiltration. Optimal conditions were achieved using detoxified concentrated hemicellulosic hydrolyzates, after CaCO₃ addition in both stages of fermentation and using nitrogen purges after sampling in order to reduce the oxygen dissolved. Under these conditions 31.5 g lactic acid L^?1 (Q_LA = 1.312 g L^?1 h^?1 and Y_LA/S = 0.93 g g^?1) and 27.5 g xylitol L^?1 (Q_Xylitol = 0.458 g L^?1 h^?1 and Y_Xylitol/S = 0.53 g g^?1) were produced. Finally, lactic acid was selectively recovered using the resin Amberlite IRA 400 (0.0374 g of lactic acid g^?1 of dry resin), allowing a further recovery of xylitol by sequential stages of adsorption, concentration, ethanol precipitation, concentration and crystallization to obtain food‐grade xylitol according to a developed process. Copyright © 2007 Society of Chemical Industry     

Firmness and Cell Wall Characteristics of Pasteurized Jalapeño Pepper Rings Affected by Calcium Chloride and Acetic Acid

Pepper rings packed in brine containing CaCl₂ were firmer, had higher bound calcium, chelator soluble pectin and pectin DE, and less water-soluble pectin (WSP) than peppers packed in brine containing no CaCl₂. Pepper rings packed in low acid brines (1% and 1.2% acetic acid) were firmer and had less WSP than those packed in high acid brine. Those samples (4% acetic acid) resulted in softening and pectin solubilization, but CaCl₂ resulted in less softening. Monosaccharide composition of cell walls was not affected by CaCl₂ or acetic acid. Firmness retention in CaCl₂ treated samples was probably due to greater association between calcium ions and pectic substances, which resisted acid hydrolysis.     

Influence of fruit ripeness and salt concentration on the microbial processing of Arbequina table olives

Arbequina table olives are processed as “naturally green olives”, they are directly placed in brine and fermentation starts spontaneously. Olives are harvested just before they change to ‘turning colour’. Different salt concentrations are used depending on the producer. The aim of the study was to evaluate how (i) the ripeness of the olive when it is harvested and (ii) the salt concentration of the brine influence the different microorganism populations in brine during the fermentation of Arbequina table olives.The results showed that the Enterobacteriaceae population lasted longer in black and turning colour olives than in green olives, whereas the growth of lactic acid bacteria was delayed in green olives. A higher salt concentration favoured the elimination of Enterobacteriaceae and hindered yeast growth. The main yeast species identified were Pichia anomala, Candida sorbosa and Candida boidinii, while Lactobacillus plantarum was the only lactic acid bacteria species involved in the process. In a sensory test, panellists preferred green olives and were not able to tell the laboratory-scale processed olives from a commercial sample, nor could they distinguish green olives from different brines.     

Effect of added glucose and xylose on the fermentation of perennial ryegrass silage inoculated with Lactobacillus plantarum

Perennial ryegrass (Lolium perenne L) was ensiled in laboratory silos after addition of glucose or xylose at rates of 0, 25, 35 and 45 g kg^?1 fresh grass. In addition, an inoculum of Lactobacillus plantarum, supplying 10⁶ organisms g^?1 fresh grass, was applied to all treatments. Silos were opened after 7, 21 and 100 days and the silage was subjected to chemical and microbiological analysis. AH silages were well fermented with pHs between 3·60 and 3·70 and low NH₃-N concentrations (<95 g kg^?1 total nitrogen) and an absence of butyric acid. Glucose was virtually completely consumed within 21 days but 0·30–0·50 of the xylose doses remained after 100 days. Lactic acid concentrations were not increased by the addition of sugars, but the glucose treatments were associated with very high concentrations of ethanol, 60–100 g kg^?1 DM, and the xylose additions produced very high concentrations of acetic acid, 60–135 g kg^?1 DM. Most(>0·80) of the glucose that disappeared could be accounted for in ethanol formation but the xylose consumed could be accounted for only if the lactic acid produced in its fermentation was metabolised further to acetic acid; indeed, for the two higher doses of xylose, the concentrations of lactic acid were reduced from the control value of 177 g kg^?1 DM to 140 and 120 g kg^?1 DM, respectively. The results indicate that the provision of extra sugar, as hexose or pentose, allows yeasts to assume a more prominent role in the fermentation with consequent wasteful fermentation of sugars. Furthermore, the suggestion is that xylose may indirectly, via a stimulation of lactate-assimilating yeasts, encourage further metabolism of lactic acid to acetic acid.     

Integral utilisation of barley husk for the production of food additives

A new and effective chemical–biotechnological process for the global utilisation of barley husk (obtained from the spent grains in the brewing process) is reported. With the proposed process the three main components of the lignocellulosic residue (cellulose, hemicellulose and lignin) are utilised. A first treatment with sulfuric acid (pre‐hydrolysis) allowed the solubilisation of hemicelluloses to give xylose and glucose‐containing liquors (suitable to make fermentation media for the continuous lactic acid (LA) production with L. pentosus) and a solid phase containing cellulose and lignin. In this set of experiments, a maximum volumetric productivity (Q_P) = 2.077 g L^?1 h^?1 and product yield (Y_P/S) = 0.62 g g^?1 were obtained for a dilution rate of 0.01 h^?1. The solid residues from pre‐hydrolysis were treated with NaOH in order to increase their cellulase digestibility, and dissolve the lignin content. The cellulose residue was used as substrates for lactic acid production by simultaneous saccharification and fermentation (SSF) in media containing Trichoderma reesei cellulases and Lactobacillus rhamnosus cells using the complete MRS broth or a cheaper medium. In both cases similar LA concentrations and volumetric productivities were achieved (P = 73.4–71.0 g L^?1 and Q_P = 1.28–1.25 g L^?1 h^?1, respectively), where P is LA concentration. The lignin solution obtained after the alkaline treatment was extracted with ethyl acetate in order to obtain the phenolic components. The extract obtained at pH 3 showed three times more antioxidant activity than the one extracted at pH 12.8, with an EC₅₀ of 1.396 g L^?1 for pH 3 and 4.604 g L^?1 for pH 12.8. The best extracts showed twice antioxidant activity than BHT. Copyright © 2007 Society of Chemical Industry