Microbial Diversity of Type I Sourdoughs Prepared and Back‐Slopped with Wholemeal and Refined Soft (Triticum aestivum) Wheat Flours

The fermentation of type I sourdough was studied for 20 d with daily back‐slopping under laboratory and artisan bakery conditions using 1 wholemeal and 2 refined soft wheat (Triticum aestivum) flours. The sourdough bacterial and yeast diversity and dynamics were investigated by plate counting and a combination of culture‐dependent and culture‐independent PCR‐DGGE approach. The pH, total titrable acidity, and concentration of key organic acids (phytic, lactic, and acetic) were measured. Three flours differed for both chemical and rheological properties. A microbial succession was observed, with the atypical sourdough species detected at day 0 (i.e. Lactococcus lactis and Leuconostoc holzapfelii/citreum group for bacteria and Candida silvae and Wickerhamomyces anomalus for yeasts) being progressively replaced by taxa more adapted to the sourdough ecosystem (Lactobacillus brevis, Lactobacillus alimentarius/paralimentarius, Saccharomyces cerevisiae). In mature sourdoughs, a notably different species composition was observed. As sourdoughs propagated with the same flour at laboratory and artisan bakery level were compared, the influence of both the substrate and the propagation environment on microbial diversity was assumed.     

Potential of amylolytic lactic acid bacteria to replace the use of malt for partial starch hydrolysis to produce African fermented pearl millet gruel fortified with groundnut

Fermentation and starch hydrolysis of a pre-cooked pearl millet–groundnut (MG) slurry inoculated with amylolytic lactic acid bacteria (ALAB) or by back slopping was investigated as a substitute for the addition of malt to prepare infant gruels. The ALAB collection strain Lb. plantarum A6, and the endogenous microflora provided by back slopping were more efficient in acidifying and partially hydrolysing starch in the MG slurry than Lb. plantarum 6.1, isolated from the traditional process in Burkina Faso. Large amounts of maltotriose and maltotetraose accumulated in slurry fermented by strain A6. No accumulation of maltose was observed, which could be an advantage to prevent the growth of microbial contaminants such as yeasts. Starch hydrolysis in the MG slurry inoculated with strain A6 or by back slopping enabled preparation of high-energy density gruels (84.7 ± 4.4 and 80.4 ± 23.8 kcal/100 g of gruel, respectively) of liquid consistency. However variability was higher with back slopping.     

Microbial characterisation and stability of a farmhouse natural fermented milk from Spain

This work reports the microbial characterisation of a farmhouse natural fermented milk (NFM) with good sensorial properties produced in Spain. Culturing and denaturing gradient gel electrophoresis (DGGE) analyses showed thatLactococcus lactissubsp.lactis and L. lactissubsp.cremoris(approximate levels of 10⁹ cfu/mL) were dominant in this NFM, whileLactobacillus plantarumappeared at a lower level (10⁶–10⁷ cfu/mL). Repetitive extragenic palindromic (REP)‐PCR typing of the isolates identified single strains each ofLc. lactissubsp.lactis, Lc. lactissubsp.cremorisandLb. plantarum. These three strains formed a stable microbial association which has been maintained for at least some decades.     

Applicability of bacteriocin‐producing Lactobacillus plantarum,Enterococcus faecium and Lactococcus lactis ssp. lactis as adjunct starter in Minas Frescal cheesemaking

The antimicrobial and technological characteristics of three bacteriocinogenic cultures used as adjunct starters in Minas Frescal cheese were investigated. The cheeses were manufactured with 1% commercial lactic starter and 0.5%Lactococcus lactis ssp. lactis ATCC 11454, Lactobacillus plantarum ALC 01 or Enterococcus faecium FAIR‐E 198. The cheeses were then artificially inoculated with Listeria monocytogenes Scott A, Staphylococcus aureus ATCC 27154 and Bacillus cereus K1‐B041 and stored for 21 days at 8°C. The results show that there was no significant difference in the counts of L. monocytogenes and S. aureus between the cheeses made with added bacteriocinogenic cultures and the control cheese. On the other hand, B. cereus exhibited susceptibility to Lb. plantarum ALC 01 and E. faecium FAIR‐E 198 from the seventh day of storage. However, these cultures increased the proteolysis of the Minas Frescal cheese.     

Comparison of PCR‐DGGE and PCR‐SSCP analysis for bacterial flora of Japanese traditional fermented fish products,aji‐narezushi and iwashi‐nukazuke

BACKGROUND: The bacterial flora of two Japanese traditional fermented fish products, aji‐narezushi (salted and long‐fermented horse mackerel (Trachurus japonicas) with rice) and iwashi‐nukazuke (salted and long‐fermented sardine (Sardinops melanostica) with rice bran), was analysed using non‐culture‐based polymerase chain reaction (PCR) denaturing gradient gel electrophoresis (DGGE) and culture‐based PCR single‐strand conformation polymorphism (SSCP) methods. RESULTS: Viable plate counts in aji‐narezushi and iwashi‐nukazuke were about 6.3–6.6 and 5.7–6.9 log colony‐forming units g^?1 respectively. In the PCR‐DGGE analysis, Lactobacillus acidipiscis was detected as the predominant bacterium in two of three aji‐narezushi samples, while Lactobacillus versmoldensis was predominant in the third sample. By the PCR‐SSCP method, Lb. acidipiscis and Lactobacillus plantarum were isolated as the predominant bacteria, while Lb. versmoldensis was not detected. The predominant bacterium in two of three iwashi‐nukazuke samples was Tetragenococcus muriaticus, while Tetragenococcus halophilus was predominant in the third sample. CONCLUSION: The results suggest that the detection of some predominant lactic acid bacteria species in fermented fish by cultivation methods is difficult. Copyright © 2010 Society of Chemical Industry     

Effect of Lactobacillus plantarum on the survival of acid‐tolerant non‐O157 Shiga toxin‐producing E. coli (STEC) strains in fermented goat's milk

The ability of goat's milk fermented with a Lactobacillus plantarum strain B411, and in combination with commercial starter culture, to inhibit acid‐adapted (AA) and non‐acid‐adapted (NAA) environmental non‐O157 STEC strains was investigated. Acid‐adapted and NAA non‐O157 STEC strains were not inhibited in the L. plantarum‐fermented goat's milk, while the goat's milk fermented with the combination of L. plantarum and starter culture inhibited AA more than NAA non‐O157 STEC strains. Environmental acid‐tolerant non‐O157 STEC strains were not inhibited by L. plantarum, starter culture or combination of starter culture with L. plantarum unless they were subjected to prior acid adaptation such as backslopping.     

Probiotic monocultures in fermented goat milk beverages – sensory quality of final product

The aim of our study was to conduct a selection of the monocultures capable of providing the most attractive sensory features of the final product. Four fermented goat's milk beverages were produced with probiotic monocultures containing Lactobacillus (Lb. acidophilus La‐5, Lb. rhamnosus K3 and Lb. plantarum O20) and Bifidobacterium (Bif. animalis subsp. lactisBB‐12). A sensory analysis and microbiological assessment of fermented goat's milk beverages were made at the beginning of the study and after 3, 7, 10 and 14 days of refrigerated storage (5 ± 1 °C). We found that samples including monocultures Lb. plantarum O20 and Bif. animalis subsp. lactisBB‐12 were differentiated from other goat's milk beverages.     

The bacteriocin producer Lactobacillus amylovorus DCE 471 is a competitive starter culture for type II sourdough fermentations

Type II sourdoughs were prepared using Lactobacillus amylovorus DCE 471, a producer of the bacteriocin amylovorin L. The strain was used as a starter culture for rye and wheat sourdoughs on laboratory scale (10 L), and in rye sourdough on pilot scale (100 L). The sourdoughs were acidified to a pH of around 3.5 within 15 h (laboratory dough) to 25 h (pilot‐scale dough). Final amylovorin L titres of 0.3–0.4 (laboratory scale) and 0.2 (pilot scale) MAU kg^?1 of sourdough were detected. After baking of wheat dough that was supplemented with the pilot‐scale sourdough, no amylovorin L activity was recovered from the breadcrumbs. On laboratory scale, aeration or the addition of complex carbohydrates hardly affected growth or amylovorin L production. Rye and wheat sourdough fermentation were rather similar despite differences in sugar concentrations. The persistence of L. amylovorus DCE 471 during rye sourdough fermentation, both on laboratory and pilot scale, was confirmed by repetitive sequence‐based polymerase chain reaction (rep‐PCR) and by testing isolates towards an amylovorin L‐sensitive organism. Further, rep‐PCR indicated that the background microbiota of the flour—probably responsible for the production of low amounts of acetic acid—grew poorly and were overgrown by L. amylovorus DCE 471 during the pilot‐scale fermentation. Copyright © 2007 Society of Chemical Industry     

Discrimination of probiotic Lactobacillus strains for poultry by repetitive sequenced‐based PCR fingerprinting

BACKGROUND: Four repetitive element sequence‐based polymerase chain reaction (rep‐PCR) methods, namely repetitive extragenic palindromic PCR (REP‐PCR), enterobacterial repetitive intergenic consensus PCR (ERIC‐PCR), polytrinucleotide (GTG)₅‐PCR and BOX‐PCR, were evaluated for the molecular differentiation of 12 probiotic Lactobacillus strains previously isolated from the gastrointestinal tract of chickens and used as a multistrain probiotic. This study represents the first analysis of the comparative efficacy of these four rep‐PCR methods and their combination (composite rep‐PCR) in the molecular typing of Lactobacillus strains based on a discriminatory index (D). RESULTS: Species‐specific and strain‐specific profiles were observed from rep‐PCR. From the numerical analysis of composite rep‐PCR, BOX‐PCR, (GTG)₅‐PCR, REP‐PCR and ERIC‐PCR, D values of 0.9118, 0.9044, 0.8897, 0.8750 and 0.8529 respectively were obtained. Composite rep‐PCR analysis was the most discriminative method, with eight Lactobacillus strains, namely L. brevis ATCC 14869^T, L. reuteri C 10, L. reuteri ATCC 23272^T, L. gallinarum ATCC 33199^T, L. salivarius ATCC 11741^T, L. salivarius I 24, L. panis JCM 11053^T and L. panis C 17, being differentiated at the strain level. CONCLUSION: Composite rep‐PCR analysis is potentially a useful fingerprinting method to discriminate probiotic Lactobacillus strains isolated from the gastrointestinal tract of chickens. Copyright © 2011 Society of Chemical Industry     

Application of Autochthonous Mixed Starter for Controlled Kedong Sufu Fermentation in Pilot Plant Tests

Traditional sufu is fermented by back‐slopping and back‐slopping has many defects. The objective of this study was to apply autochthonous mixed starter to control Kedong sufu fermentation. Sufu was manufactured using back‐slopping (batch A) and autochthonous mixed starter (batch B) with Kocuria kristinae F7, Micrococcus luteus KDF1, and Staphylococcus carnosus KDFR1676. Considering physicochemical properties of sufu, 150‐day sufu samples from batch A and 90‐day sufu samples from batch B met the standard requirements, respectively. Considering sensory characteristics of sufu, 150‐day sufu samples from batch A and 90‐day sufu samples from batch B showed no significant differences (P > 0.05). The maturation period of sufu was shortened by 60 d. Profiles of free amino acids and peptides partly revealed the mechanism of typical sensory quality and shorter ripening time of sufu manufactured by autochthonous mixed starter. In final products, content of total biogenic amines was reduced by 48%. Autochthonous mixed starter performed better than back‐slopping. Fermentation had a positive influence on the quality, safety, and sensory properties of sufu. The application of autochthonous mixed starter does not change the sensory characteristics of traditional fermented sufu. In addition, it reduces maturation period and improves their homogeneity and safety. It is possible to substitute autochthonous mixed starter for back‐slopping in the manufacture of sufu.     

A centrifugation-based clearing method allows high-throughput acidification and growth-rate measurements in milk

The turbidity of milk prohibits the use of optical density measurements for strain characterizations. This often limits research to laboratory media. Here, we cleared milk through centrifugation to remove insoluble milk solids. This resulted in a clear liquid phase, termed milk serum, in which optical density measurements can be used to track microbial growth until a pH of 5.2 is reached. At pH 5.2 coagulation of the soluble protein occurs, making the medium opaque again. We found that behavior in milk serum was predictive of that in milk for 39 Lactococcus lactis (R² = 0.81) and to a lesser extent for 42 Lactiplantibacillus plantarum (formerly Lactobacillus plantarum; R² = 0.49) strains. Hence, milk serum can be used as an optically clear alternative to milk for comparison of microbial growth and metabolic characteristics. Characterization of the growth rate, specific acidification rate for optical density at a wavelength of 600 nm, and the amount of acid produced per unit of biomass for all these strains in milk serum, showed that almost all strains could grow in milk, with higher specific acidification and growth rates of Lc. lactis strains compared with Lb. plantarum strains. Nondairy Lc. lactis isolates had a lower growth and specific acidification rate than dairy isolates. The amount of acid produced per unit biomass was relatively high and similar for Lc. lactis dairy and nondairy isolates, as opposed to Lb. plantarum isolates. Lactococcus lactis ssp. lactis showed slightly lower growth and acidification rates when compared with ssp. cremoris. For Lc. lactis strains a doubling of the specific acidification rate occurred with a doubling of the maximum growth rate. This relation was not found for Lb. plantarum strains, where the acidification rate remained relatively constant across 39 strains with growth rates ranging from 0.2 h^?1 to 0.3 h^?1. We conclude that milk serum is a valuable alternative to milk for high-throughput strain characterization during milk fermentation.     

Description of the microflora of sourdoughs by culture-dependent and culture-independent methods

Four types of sourdoughs (L, C, B, Q) from artisanal bakeries in Northern Italy were studied using culture-dependent and culture-independent methods. In all samples, the yeast numbers ranged from 160 to 10⁷ cfu/g, and the numbers of lactic acid bacteria (LAB) ranged from 10³ to 10⁹ cfu/g. The isolated LAB were sequenced, and a similarity was noted between two samples (C, Q), both in terms of the species that were present and in terms of the percentage of isolates. In these two samples, Lactobacillus plantarum accounted for 73% and 89% of the bacteria, and Lactobacillus brevis represented 27% and 11%. In the third sample (B), however, the dominant LAB isolate was Lb. brevis (73%), while Lb. plantarum accounted for only 27%. The fourth sourdough (L) was completely different from the others. In this sample, the most prominent isolate was Weisella cibaria (56%), followed by Lb. plantarum (36%) and Pediococcus pentosaceus (8%). In three out of four samples (L, C and Q), all of the yeasts isolated were identified as Saccharomyces cerevisiae, yet only Candida humilis (90%) and Candida milleri (10%) were isolated in the fourth sample (B). The microbial ecology of the sourdoughs was also examined with direct methods. The results obtained by culture-independent methods and DGGE analysis underline a partial correspondence between the DNA and RNA analysis. These results demonstrate the importance of using a combined analytical approach to explore the microbial communities of sourdoughs.     

Comparative study of spontaneously fermented sourdoughs originating from two regions of Greece: Peloponnesus and Thessaly

A total of 167 yeast and 136 lactic acid bacteria strains were isolated from spontaneously fermented wheat sourdoughs from two regions of Greece, namely Thessaly and Peloponnesus. Identification of the isolates exhibited dominance of Torulaspora delbrueckii with sporadic presence of Saccharomyces cerevisiae and, regarding the lactic acid bacteria, dominance of Lactobacillus sanfranciscensis in the sourdoughs from Thessaly and Lb. plantarum subsp. plantarum in the sourdoughs from Peloponnesus. The latter was accompanied by Pediococcus pentosaceus as secondary microbiota. None of the above mentioned strains exhibited amylolytic, lipolytic or proteolytic activities, and none of the lactic acid bacteria strains produced antimicrobial compounds.     

Sourdough-isolated<Emphasis Type="Italic"> Lactobacillus fermentum</Emphasis> as a potent anti-mould preservative of a traditional Iranian bread

Sourdough bread is an ancient method for making long microbial shelf life and strongly flavoured breads. Traditional sourdough bread-making has been practised for centuries in Iran. Although rural bread-making is still highly reliant on sourdough, urban bakeries usually use bakers yeast and also sodium bicarbonate instead. In this study the anti-mould preservative effects of three lactic acid bacteria isolates were investigated on one of the popular traditional Iranian bread (Lavash). Starter cultures were prepared using 20 h cultures of three sourdough-isolated strains, Lactobacillus casei, Lactobacillus plantarum and Lactobacillus fermentum. Different concentrations (0.5%, 1%, 2%, 4% and 8%) of lactobacillus starter cultures were used to prepare a variety of sourdoughs. Lavash doughs were inoculated with 20% of each lactobacilli-fermented sourdough. After a 20 h incubation at 30 °C, a decrease in pH was observed in the different lactobacilli sourdoughs. However an 8% inoculum of L. fermentum resulted in a significant decrease in pH (3.87 to 2.70). Concentrations of 2% and higher of the three lactobacilli used in Lavash sourdough delayed mould growth during storage. However this preservative effect was more significant when an 8% inoculum of L. fermentum was used. These results tend to suggest that selected strains of lactobacilli have a beneficial role in prevention of bread waste by mould spoilage, and hence could extend bread shelf life.     

Effect of Select Parameters of the Sourdough Rye Fermentation on the Activity of Some Mixed Starter Cultures

The effect of select parameters (i.e., rye flour ash content, temperature, and dough yield) of the sourdough fermentation on the fermentation activity of different starter cultures (lactic acid bacteria Lactococcus lactis ssp. Lactis, Weissella confusa, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus helveticus, and yeast Kluyveromyces marxianus subsp. Marxianus) was determined. The major metabolic end products of fermentation (D, L-lactic acid, acetic acid, ethanol and glycerol) and the evolution of total phenolic content and folic acid during bread making were measured. Lactobacillus helveticus and Kluyveromyces marxianus allowed obtaining sourdoughs with the highest lactic acid/acetic acid ratios. The mixed starter culture with Lactococcus lactis and Saccharomyces cerevisiae generated the most important quantities of D/L lactic acid. The maximum values of ethanol concentration were obtained in case of the sourdoughs from whole rye flour fermented at lower temperature (30°C) with mixed starter cultures containing Sacchomyces cerevisiae. The fermentation process and type of starter culture are also tools to increase the bioactive compounds, enabling the increase of the phenolic content of the sourdough.     

Ability of Selected Lactic Acid Bacteria to Ferment a Pearl Millet–Soybean Slurry to Produce Gruels for Complementary Foods for Young Children

Abstract: To assess the ability of lactic acid bacteria to improve some nutritional characteristics of the pearl millet–soybean slurry to prepare complementary foods for young children in African countries, inoculation was performed using strains previously selected for their ability to hydrolyse starch, phytate, or α-galactooligosaccharides (α-GOS). For the sake of comparison with the action of a natural microflora, fermentation was also performed by back slopping inoculation, that is, with a sample obtained from spontaneously fermented traditional pearl millet slurry obtained from a small scale processing unit in Burkina Faso (Ouagadougou). Starter cultures thrived on the slurry as shown by counts on MRS agar, TTGE fingerprints, and fermentation patterns. The fermentation of precooked slurries inoculated by back slopping or with mixed cultures containing the amylolytic strain Lb. plantarum A6 enabled partial starch hydrolysis. Corresponding gruels had a suitable consistency for young child feeding at high dry matter content, and a high energy density: 88.7 ± 4.2 and 75.8 ± 5.1 kcal/100 g of sweetened gruel, for the gruels inoculated by back slopping or with Lb. plantarum A6, respectively. Unexpectedly, no decrease in phytates was observed in any of the experiments, suggesting the presence of one or more inhibitory compounds in soybean. Furthermore, preprocessing conditions before fermentation affect the carbohydrate composition of slurry and have a more profound effect than fermentation on the reduction of the α-GOS content. Practical Application: This research aims to investigate the capacity of selected lactic acid bacteria to produce in a bioprocess high energy density and nutritious gruels for complementary feeding of young children from an African cereal (pearl millet) mixed with soybean.     

The effect of using an exopolysaccharide‐producing culture on the physicochemical properties of low‐fat and reduced‐fat Kasar cheeses

A mixed starter culture containing exopolysaccharide (EPS)‐producing strains of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was combined with Lactobacillus helveticus LH301 and used in the manufacture of low‐fat and reduced‐fat Kasar cheeses. For comparison, low‐fat (C10) and reduced‐fat (C20) cheeses were made using EPS‐producing (EPS⁺) starter strain and EPS‐non‐producing (EPS^?) starter strain. The physicochemical properties of the cheeses were assessed in terms of chemical composition, texture, microstructure and microbial content over 90 days. Cheeses made with EPS‐producing culture (EPS10 and EPS20) had lower protein contents than control cheeses with 10% and 20% fat in dry basis (C10 and C20). Scanning electron microscopy images showed that using EPS‐producing culture resulted in a less compact protein matrix and sponge‐like structure in the cheese samples. In general, cheeses made using EPS‐producing culture had lower total viable counts. This could be related to the reduced survivability of EPS‐producing cells in the cheese matrix during ripening due to autolysis ability.     

Preliminary observations on proteolysis in Manchego cheese made with a defined-strain starter culture and adjunct starter (Lactobacillus plantarum) or a commercial starter

Different authors have demonstrated the potential of adding lactobacilli as adjunct cultures to pasteurized milk used in cheese manufacture. The aim of this work was to observe the effect of the use of a defined-strain starter culture and the addition of an adjunct culture (Lactobacillus plantarum) to cheesemilk in order to determine their effect on the ripening of Manchego cheese. Manchego cheeses were manufactured using one of the following starter culture systems: a defined starter consisting of Lactococcus lactis ssp. lactis and Leuconostoc mesenteroides ssp. dextranicum; a defined starter, as described above, and Lb. plantarum, which were isolated from a good quality Manchego cheese made from raw milk, or a commercial starter comprised of two strains of Lc. lactis. The cheeses were sampled at 15, 45, 90 and 150 d of ripening. Principal component analysis of peak heights of reversed-phase HPLC chromatograms of 70% (v/v) ethanol-insoluble and -soluble fractions distributed the samples according to the starter used in their manufacture. Quantitative differences in several peptides were evident between the three cheeses. Cheeses made with the defined-strain starters received higher scores for the flavour quality and intensity and for overall impression than the cheeses made with the commercial starter.     

Application of Different Molecular Techniques for Characterization of Catalase‐Positive Cocci Isolated from Sucuk

This study was carried out for the characterization and discrimination of the indigenous Gram positive, catalase‐positive cocci (GCC) population in sucuk, a traditional Turkish dry‐fermented sausage. Sucuk samples, produced by the traditional method without starter culture were collected from 8 local producers in Kayseri/Turkey and a total of 116 GCC isolates were identified by using different molecular techniques. Two different molecular fingerprinting methods; namely, randomly amplified polymorphic DNA‐PCR (RAPD‐PCR) and repetitive extragenic palindrome‐PCR (rep‐PCR), were used for the clustering of isolates and identification at species level was carried out by full length sequencing of 16S rDNA. Combining the results obtained from molecular fingerprinting and 16S rDNA sequencing showed that the dominant GCC species isolated from the sucuk samples was Staphylococcus saprophyticus followed by Staphylococcus succinus and Staphylococcus equorum belonging to the Staphylococcus genus. Real‐time PCR DNA melting curve analysis and high‐resolution melting (HRM) analysis targeting the V1 + V3 regions of 16S rDNA were also applied for the discrimination of isolates belonging to different species. It was observed statistically different Tm values and species‐specific HRM profiles for all except 2 species (S. saprophyticus and Staphylococcus xylosus) that have high 16S rDNA sequence similarity. The combination of rep‐PCR and/or PCR‐RAPD with 16S rRNA gene sequencing was an efficient approach for the characterization and identification of the GCC population in spontaneously fermented sucuk. On the other hand, intercalating dye assays were found to be a simple and very promising technique for the differentiation of the GCC population at species level.     

Investigation of Microbial Communities of Chinese Sourdoughs Using Culture‐Dependent and DGGE Approaches

Sourdough is typically characterized by the complex microbial communities mainly comprising of yeasts and lactic acid bacteria (LAB). The objective of this study was to explore the microbiota of Chinese traditional sourdoughs collected from different areas of China using culture‐dependent and denaturing gradient gel electrophoresis (DGGE) methods. A total of 131 yeasts, 2 molds, and 106 LAB strains were isolated and identified. Based on the culture‐dependent analysis, the populations of yeasts and LAB were at the level of 10⁵ to 10⁷ and 10⁶ to 10⁷ cfu/g, respectively. Similarly, the results of RT‐qPCR showed that the values of total yeasts and LAB populations were in the range of 10⁶ to 10⁷ and 10⁷ to 10⁸ copies/g, respectively. Using culture‐dependent method, a total of 7 yeasts, 2 molds and 7 LAB species were identified. Results showed that Saccharomyces cerevisiae and Lactobacillus plantarum were the predominant species among the yeasts and LAB microflora. Similarly, using PCR‐DGGE approach, 7 yeasts, 1 mold and 9 LAB species were detected. The yeast, S. cerevisiae, represented the predominant, while the yeast Candida tropicalis represented the subdominant species of the yeast community. Among the LAB community, Lactobacillus sanfranciscensis was the predominant species, while Lactococcus qarvieae, Enterococcus faecium, Lactobacillus delbrueckii and Enterococcus cecorum were among the less dominant species.