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.     

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.     

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.     

Characterization of lactic acid bacteria isolated from Italian Bella di Cerignola table olives: selection of potential multifunctional starter cultures

Lactic acid bacteria (19 isolates) from Bella di Cerignola Italian table olives were investigated for their technological and probiotic properties for the selection of multifunctional starter cultures for table olives. The bacteria were first identified by phenotyping and genotyping, then characterized for the production of biogenic amines, growth at different pH, NaCl concentrations, and temperatures. The potentiality of the bacteria to have some probiotic properties (antimicrobial activity against foodborne pathogens, survival in low pH and in the presence of bile salts, ability to adhere to the mammalian cells model IPEC-J2) was also investigated. Eighteen of the studied isolates were identified as Lactobacillus plantarum and one as Enterococcus faecalis. All bacteria were able to grow at a range of pH between 4.0 and 10.0 as well as in media supplemented with 2.5 to 7.5% of NaCl and 0.3% bile salts and survived in MRS broth acidified at pH 2.5; moreover, they inhibited significantly Escherichia coli O157:H7. The adhesion to IPEC-J2 cells was in general low to moderate (5.3 to 8.3%); however, 2 isolates of L. plantarum (c16 and c19) showed interesting higher adhesion values (up to 16%). Our results suggest that at least 3 isolates could be possible multifunctional starters for Bella di Cerignola olives: L. plantarum 16 and 19 for mainly their probiotic properties and L. plantarum 10 for mainly its technological characteristics. Practical Application: A functional starter is a microorganism exerting benefits on human health (probiotic) and able to guide a fermentation (starter). The main goal of this article was to select a functional starter for table olives.     

Identification and characterization of yeast isolated from the elaboration of seasoned green table olives

The purpose of this study was to investigate the yeast population during the processing of green table olives. In the fresh olives, yeast were found at concentrations of around 3.0 log cfu/g, with Cryptococcus spp. being predominant. In the brine, the yeast concentrations were greater than 4.9 log cfu/ml, with Pichia anomala, Kluyveromyces marxianus, and Saccharomyces cerevisiae being the predominant species. Unlike the yeast isolated from the fresh olives, the strains obtained from the olive brine mostly showed low pectolytic but high catalase activities. Some of these strains also exhibited other biochemical desirable properties for the fermentation of green table olives, including their lipolytic activities and their assimilation or production of organic acids in the brine. Seven strains in particular of P. anomala, K. marxianus, S. cerevisiae, and Candida maris showed the best properties for use in trials as starter culture in pilot fermenters.     

Effects of post-fermentation processing on the stabilisation of naturally fermented green table olives (cv Nocellara etnea)

The effects of thermal treatment on green table olives were evaluated as a method to control enzymatic browning, to minimise the microorganism presence and to extend their shelf-life. However this treatment is often responsible for colour alterations, development of off-flavours and unfavourable texture changes. Moreover, the effect of different re-use of the natural brine of fermentation with or without different treatments was investigated. Calcium treatment was suggested to maintain firmness. Firming effects obtained from heat treatment combined with calcium treatment have been attributed to heat-activated pectin methylesterase and/or to increased calcium diffusion into tissues at higher temperatures. The results derived from this study will help in designing new processes which can be applied in table olive industry.     

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.     

Yeast dynamics during the fermentation of brined green olives treated in the field with kaolin and Bordeaux mixture to control the olive fruit fly

The yeast microbiota associated with naturally fermented and inoculated green table olives, differently treated in the field with non-conventional repellent and antiovipositional products in the control of Bactrocera oleae, was analysed using a combination of culture-dependent and -independent molecular fingerprinting. The routine yeast isolation gave rise to 118 strains, whose identification was performed by PCR-RFLP of the internal transcribed spacer (ITS) regions. Total DNA was extracted directly from the brine throughout fermentation by means of an experimental protocol that included the removal of Taq polymerase inhibitors. Denaturing Gradient Gel Electrophoresis (DGGE) of 26S rRNA gene PCR amplicons highlighted the yeast community. Comparison of both culture-dependent and independent methods indicated that the yeast species Saccharomyces cerevisiae, Wickerhamomyces anomalus, Candida diddensiae and Issatchenkia orientalis were dominant during fermentation despite the addition of the Lactobacillus plantarum starter used in brining. The resultant isolated species were unaffected by treatments in field, except for C. diddensiae whose growth was delayed by kaolin.     

Influence of starter cultures on the antioxidant activity of kombucha beverage

This paper investigates the influence of starter cultures, obtained from kombucha isolates, on the antioxidant activity of kombucha beverages. Three starter cultures were used as follows: (1) mixed culture of acetic bacteria and Zygosaccharomyces sp. (SC1); (2) mixed culture of acetic bacteria and Saccharomyces cerevisiae (SC2); as well as (3) native local kombucha. The starter cultures were added to black and green tea sweetened with 7% of sucrose. Fermentation was carried out at 28 °C for 10 days. Antioxidant activity to hydroxyl and DPPH radicals was monitored. Kombucha beverage on black tea has shown the highest antioxidant activity to both types of radicals with starter SC1, while the green tea beverage has shown the highest activity with native kombucha. The main reason for the different antioxidant activities, beside tea composition, was ascribed to differing production of both vitamin C and total organic acids in the investigated systems.     

Changes in the volatile composition of Spanish-style green table olives induced by pasteurisation treatment

Thermal treatment of pasteurisation is the most widely used stabilisation method for packed green table olives. In the present work, the influence of pasteurisation on the composition of volatile compounds in packed Spanish-style green table olives was studied. To this aim, two thermal processes (P1 = 85 °C for 7 min; P2 = 85 °C for 15 min) were evaluated by comparing the contents of volatile compounds in pasteurised olives with those in unpasteurised olives. Volatiles were analysed, both in olive juice and cover brine, by means of headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS). Both pasteurisation treatments caused significant increases in diverse ethyl and methyl esters and in volatile compounds derived from several chemical reactions (lipid oxidation, Maillard reaction, degradation of carotenoids). Significant differences between samples subjected to P1 and P2 were only found for 1-octen-3-ol, pentanal and 6-methyl-5-hepten-2-one in olive juice.     

Bioactivity of tempe by inhibiting adhesion of ETEC to intestinal cells,as influenced by fermentation substrates and starter pure cultures

Soya bean tempe is known for its bioactivity in reducing the severity of diarrhoea in piglets. This bioactivity is caused by an inhibition of the adhesion of enterotoxigenic Escherichia coli (ETEC) to intestinal cells. In this paper, we assessed the bioactive effect of soya tempe on a range of ETEC target strains, as well as the effect of a range of cereal and leguminous substrates and starter pure cultures.     

Influence of starter cultures on the free fatty acids during ripening in Tea sausages

The influence of starter cultures on the free fatty acids content during ripening of Tea sausages, typical dry sausages produced in the south of Croatia, was studied. Three batches of Tea sausages were produced using different starter mixtures (Staphylococcus xylosus S81 and Lactobacillus sakei G20; Staphylococcus xylosus S142 and Lactobacillus sakei G20; Staphylococcus xylosus S206 and Lactobacillus sakei G20), while the control batch was produced without a starter. The amounts of free fatty acids present in the samples at the end of the ripening period were not significantly different, suggesting that the lipolytic enzymes naturally occurring in meat could play a predominant role in the free fatty acids release. Oleic and linoleic acids were present in the highest concentrations, while only small quantities of short-chain fatty acids were detected, with acetic acid being the most representative one.     

Effects of starter cultures and nitrite levels on formation of biogenic amines in sucuk

The effects of starter cultures, starter A (Lactobacillus sakei+Staphylococcus carnosus) and starter B (Pediococcus acidilactici+Staphylococcus xylosus+Lactobacillus curvatus), nitrite levels (0, 75, and 150ppm) and ripening period on biogenic amine production were investigated in sucuk (Turkish dry-fermented sausage) production. Changes in biogenic amines, pH, water activity, moisture, non-protein nitrogen, nitrite and counts of lactic acid bacteria, Micrococcus/Staphylococcus, Enterobacteriaceae, and moulds-yeasts were determined. Ripening period had a very significant effect (P<0.01) on the all biogenic amines. Sausages produced by fermentation with starter cultures, as compared to natural fermentation (control), had lower amounts of putrescine, cadaverine and tyramine (P<0.05). Starter cultures A and B were found to have almost the same effect on the formation of biogenic amines. Use of nitrite in sucuk production affected the formation of biogenic amines except for spermidine and spermine (P<0.01). Lactic acid bacteria counts in the control group were lower than that of starter culture groups. Lactic acid bacteria, Micrococcus/Staphylococcus, and moulds-yeasts counts increased during ripening period while levels of species of Enterobacteriaceae decreased, which was also found to be under the detectable level (<100cfug(-1)), by day 3 of ripening.     

Type of bacterial starter culture,aging and fermentation effects on some characteristics of inoculated beef sausages

Beef sausage mixes were inoculated with either Pediococcus acidilactici with Staphylococcus xylosus or P. acidilactici with S. carnosus, subdivided and then held for 0, 24, 48 or 72 h at 8–10 °C prior to fermentation. After aging (pre-fermentation holding), the mixes were fermented for 16 h ending at 41 °C. Moisture, protein and fat contents of all sausage mixes did not differ due to holding effects over all starter cultures. The pH of mixes followed the same pattern for all mixes, declining (p < 0.05) from approximately 5.8 to pH 5.2–5.3 at 72 h aging and to 4.4–4.5 after fermentation. Total acidity of the mixes followed an inverse pattern to pH, increasing (p < 0.05) after fermentation although there was no effect due to type of starter culture. Aging had no effect on nonprotein nitrogen (NPN) content as ΔNPN among all cultures. After fermentation, however, sausages held 72 h and inoculated with S. carnosus had higher NPN contents compared to P. acidilactici alone (p < 0.05) and with S. xylosus (p < 0.10). The same effects of starter cultures on changes in total amino acid concentration were observed. Concentrations of individual amino acids showed increases depending on pre-fermentation aging period (0 h versus 72 h) followed by fermentation.     

Proteolytic and lipolytic starter cultures and their effect on traditional fermented sausages ripening and sensory traits

In this study, three starter formulations including Lactobacillus curvatus and Staphylococcus xylosus strains selected in vitro on the basis of their lipolytic and proteolytic activities were employed for the manufacture of traditional fermented sausages of southern Italy. Microbial population, proteolysis, lipolysis, changes in free amino acids (FAA) and free fatty acids (FFA) and development of characteristic taste and flavor of the final product were investigated. Proteolysis and lipolysis were observed in sausages inoculated with proteolytic and lipolytic S. xylosus coupled with L. curvatus, while the sausage started with only S. xylosus without lactobacilli was identical to the non-inoculated control, indicating that the proteolysis could be due to both microbial activity and endogenous proteases activated by the decrease in pH. The statistical analysis applied to the instrumental and sensory data showed that there was an effect of the starter used on the characteristics of the sausage obtained. In particular, the control samples showed very close features different from the sausages obtained by adding starter cultures. Finally, analyzing the sensory parameters the sausages ripened without starter addition and those started without the L. curvatus AVL3 showed similar features indicating an influence of the presence of the lactobacilli on the final organoleptic quality of the sausages. An appropriate choice of a combination of strains in a starter formulation is fundamental to obtain products of the expected quality.     

乳酸菌肉品发酵剂的发酵特性研究

对从湘西泡菜水中分离出的5株植物乳杆菌（Lactobacillus planterum）,1株肠系膜明串珠菌（Leuconostocmesenteroides）,1株戊糖片球菌（Pediococcuspentosaceus）,进行耐盐和亚硝酸盐能力、生长和产酸能力、硝酸盐还原和氨基酸脱羧能力的研究以及一系列生理生化实验来验证其在肉品发酵上的可用性。结果表明:植物乳杆菌（Lactobacillus planterum-A）和戊糖片球菌（Pediococcuspentosaceus-6）对食盐和亚硝酸盐均具有较好的耐受性,对蛋白质和脂肪不具有明显的分解作用,不具有硝酸盐还原酶和氨基酸脱羧酶活性,对大肠杆菌（Escherichia coli）,沙门氏菌（Salmonellas spp.）等实验菌株有较好的抑制作用;70℃可致死。符合作为肉品发酵剂的要求,可作为下步实验的出发菌株。      

Prevalence and impact of single-strain starter cultures of lactic acid bacteria on metabolite formation in sourdough

Flavour of type II sourdoughs is influenced by the ingredients, processing conditions, and starter culture composition. It is, however, not fully clear to what extent different sourdough lactic acid bacteria (LAB) contribute to flavour. Therefore, two types of flour (rye and wheat) and different LAB starter culture strains were used to prepare sourdoughs, thereby leaving the yeast microbiota uncontrolled. All LAB starter culture strains tested were shown to be prevalent and to acidify the flour/water mixture to pH values between 3.1 and 3.9 after 24 h of fermentation. Multiple aldehydes, alcohols, ketones, and carboxylic acids were produced by the sourdough-associated microbiota throughout the fermentation period. Based on the organoleptic evaluation of breads produced with these sourdoughs, five LAB strains were selected to perform prolonged wheat and rye fermentations as to their capacity to result in an acidic (Lactobacillus fermentum IMDO 130101, Lactobacillus plantarum IMDO 130201, and Lactobacillus crustorum LMG 23699), buttermilk-like (Lactobacillus amylovorus DCE 471), or fruity flavour (Lactobacillus sakei CG1). Upon prolonged fermentation, higher metabolite concentrations were produced. For instance, L. sakei CG1 produced the highest amounts of 3-methyl-1-butanol, which was further converted into 3-methylbutyl acetate. The latter compound resulted in a fruity banana flavour after 48 h of fermentation, probably due to yeast interference. Rye fermentations resulted in sourdoughs richer in volatiles than wheat, including 3-methyl-1-butanol, 2-phenylethanol, and ethyl acetate.     

开菲尔功能菌组的复合及发酵特性研究

研究了开菲尔粒子中不同分离菌种的发酵情况,对不同比例的乳酸菌、酵母发酵产物的质量指标(如风味、酸度和发泡性)进行了评价,不同比例分离菌种(30～50g/L)在均质乳中(30℃,16h)连续发酵培养,分析检测其中的蛋白质、脂肪、粗多糖、酒精、黏度和VB含量。结果表明,L-1∶L-2∶L-3∶L-4∶L-5∶Y-1∶Y-2=1∶1∶1∶1∶1∶1发酵产物经评价其质量和感官特性最接近开菲尔。     

Utilization of Enterococcus casseliflavus and Lactobacillus pentosus as starter cultures for Spanish-style green olive fermentation

Green olive were fermentated with starter cultures of Enterococcus casseliflavus cc45 andLactobacillus pentosus CECT 5138. A total of 32 strains of enterococci isolated from brines at high pH were characterized and identified by phenotypic methods. The strain cc45 was selected after a preliminary screening and applied together with thelactobacillus culture the first day after brining, comparing the results with those obtained when both cultures were used sequentially, enterococci at day 1 and lactobacilli at day 2, and with uninoculated controls. Microbial growth, substrate and product evolution, and flavour of the fermented olives were studied. Inoculation produced a quicker acidification of brines, consumption of carbohydrates and decrease of pH. The best results were attained when both cultures of lactic acid bacteria were applied sequentially with 1day delay. This is the first report dealing with the presence of E. casseliflavus in table olive brines and their utilization as starter culture.     

发酵剂对熏马肠脂肪酶活力的影响

主要研究发酵剂对熏马肠发酵成熟过程中脂肪酶活力的影响。结果表明,酸性脂肪酶活力、中性脂肪酶活力、磷脂酶活力在发酵成熟过程中持续下降,并且发酵剂组的三种酶活力在添加发酵剂后显著高于空白组(p0.05)。通过相关性分析表明,在熏马肠发酵成熟过程中,发酵剂组中的三种酶活力与p H、水分含量及盐分含量呈极显著相关性(p0.01)。