Exploitation of Selected Sourdough Saccharomyces cerevisiae Strains for the Production of a Craft Raspberry Fruit Beer.

Recent interest in the special beer category has encouraged the search for novel brewing materials, including new ingredients and novel yeast strains, in order to differentiate the finished products. The aim of this work was to select non-brewing S. cerevisiae strains for the production of a fruit beer with raspberry. The in vitro tests and the wort fermentations allowed the selection of two sourdough S. cerevisiae strains, showing high maltose and maltotriose consumption, high ethanol production, and high viability. Fruit beers (FB) and control beers (CB) without raspberries were prepared. Fruit addition accelerated sugar consumption (7 days compared to 13 days) and increased ethanol and glycerol production by yeasts. Raspberry addition and the inoculated yeast strongly affected the aroma profile of beers. FB samples showed a higher amount of volatile organic compounds (VOCs); the most represented classes were alcohols, followed by esters and acids. FB inoculated by the selected S. cerevisiae SD12 showed the highest VOCs concentration (507.33 mg/L). Results highlighted the possible application of sourdough yeast strains for the brewing process, which, combined with raspberry addition, can be exploited for the production of beers with enhanced aromatic features and suitable chemical properties.

Management of in-Amphora "Trebbiano Toscano" Wine Production: Selection of Indigenous Saccharomyces cerevisiae Strains and Influence on the Phenolic and Sensory Profile.

The use of earthenware amphorae in winemaking can give wines unique attributes enhancing their typicity. Therefore, in this study, spontaneous and inoculated in-amphora fermentations of Trebbiano Toscano grape must were monitored to assess the Saccharomyces cerevisiae strains occurring in each fermentation as well as the chemical characteristics of the wines. Strain typing via Interdelta analyses pointed out that the commercial starters did not dominate, showing 24% and 13% implantation percentages, and that 20 indigenous strains were present at different percentages, ranging from 2 to 20%, in inoculated and spontaneous fermentations. The assessment of the technical characteristics of the indigenous strains via fermentations at lab and pilot scale (20 L amphorae) and the sensory analysis of the experimental wines allowed for the selection of two indigenous strains to be used as starter cultures in comparison to a commercial strain in 300-L-amphorae vinifications in the cellar. The observed fermentative performances and sensory analysis of the experimental wines highlighted that one indigenous S. cerevisiae strain dominated the process and conferred distinctive sensory characteristics to the Trebbiano Toscano wine, demonstrating its effectiveness in managing the in-amphora fermentations. In addition, the results demonstrated the ability of amphorae to protect the polyphenolic compounds from oxidation during wine ageing. Indeed, the concentration of both hydroxycinnamic acids and flavonols decreased, with an average reduction of 30% and 14%, respectively, while hydroxybenzoic acids remained unchanged.

Replacement of SO2 with an Unripe Grape Extract and Chitosan during Oak Aging: Case Study of a Sangiovese Wine.

The aim of this study was to evaluate the chemical, microbiological and sensory characteristics of a Sangiovese wine aged in barrique with the addition of an unripe grape extract (UGE) as an alternative to sulfur dioxide. Three samples were considered: control wine (TQ) with free SO2 of approximately 15 mg/L; sample A with chitosan (100 mg/L) and UGE (200 mg/L); and sample B with UGE (400 mg/L). The results achieved in this work demonstrated that the UGE, either alone or in combination with chitosan, was able to maintain the color characteristics of the Sangiovese wine and its sensory quality. Moreover, the addition of UGE contributed to an early and better stabilization of the color through the formation of polymeric pigments. The microbiological stabilization was comparable to SO2 when UGE was used at 200 mg/L in combination with chitosan. The market survey conducted in the present study confirmed how the use of UGE as an alternative to sulfitation was positively accepted by consumers, who are increasingly attentive not only to the quality of the wines they select but also to the sustainability of the production processes from which they derive and to the fact that they are not harmful to human health.

Rapid Detection of Brettanomyces bruxellensis in Wine by Polychromatic Flow Cytometry.

Brettanomyces bruxellensis is found in several fermented matrices and produces relevant alterations to the wine quality. The methods usually used to identify B. bruxellensis contamination are based on conventional microbiological techniques that require long procedures (15 days), causing the yeast to spread in the meantime. Recently, a flow cytometry kit for the rapid detection (1-2 h) of B. bruxellensis in wine has been developed. The feasibility of the method was assessed in a synthetic medium as well as in wine samples by detecting B. bruxellensis in the presence of other yeast species (Saccharomyces cerevisiae and Pichia spp.) and at the concentrations that produce natural contaminations (up to 105 cells/mL), as well as at lower concentrations (103-102 cells/mL). Wine samples naturally contaminated by B. bruxellensis or inoculated with four different strains of B. bruxellensis species together with Saccharomyces cerevisiae and Pichia spp., were analyzed by flow cytometry. Plate counts were carried out in parallel to flow cytometry. We provide evidence that flow cytometry allows the rapid detection of B. bruxellensis in simple and complex mixtures. Therefore, this technique has great potential for the detection of B. bruxellensis and could allow preventive actions to reduce wine spoilage.

Validation of a Standard Protocol to Assess the Fermentative and Chemical Properties of Saccharomyces cerevisiae Wine Strains.

This paper reports on a common experiment performed by 17 Research Units of the Italian Group of Microbiology of Vine and Wine (GMVV), which belongs to the Scientific Society SIMTREA, with the aim to validate a protocol for the characterization of wine strains of Saccharomyces cerevisiae. For this purpose, two commercial S. cerevisiae strains (EC 1118 and AWRI796) were used to carry out inter-laboratory-scale comparative fermentations using both synthetic medium and grape musts and applying the same protocol to obtain reproducible, replicable, and statistically valid results. Ethanol yield, production of acetic acid, glycerol, higher alcohols, and other volatile compounds were assessed. Moreover, the Fourier transform infrared spectroscopy was also applied to define the metabolomic fingerprint of yeast cells from each experimental trial. Data were standardized as unit of compounds or yield per gram of sugar (glucose and fructose) consumed throughout fermentation, and analyzed through parametric and non-parametric tests, and multivariate approaches (cluster analysis, two-way joining, and principal component analysis). The results of experiments carried out by using synthetic must showed that it was possible to gain comparable results from three different laboratories by using the same strains. Then, the use of the standardized protocol on different grape musts allowed pointing out the goodness and the reproducibility of the method; it showed the main traits of the two yeast strains and allowed reducing variability amongst independent batches (biological replicates) to acceptable levels. In conclusion, the findings of this collaborative study contributed to the validation of a protocol in a specific synthetic medium and in grape must and showed how data should be treated to gain reproducible and robust results, which could allow direct comparison of the experimental data obtained during the characterization of wine yeasts carried out by different research laboratories.

Characterization of the microbial community in ripened Pecorino Toscano cheese affected by pink discoloration.

Pink discoloration defect can cause economic losses for cheese producers due to the impossibility to sell the defected cheese, but few knowledge is currently available on the causes of this defect. To gain more insight on the causes that lead to the formation of pink discoloration in Pecorino Toscano cheese with the Protected Designation of Origin (PDO) status, the bacterial community in defected and not defected cheese was characterized by high-throughput sequencing of bacterial 16S rRNA gene. The bacterial community in the defected cheese significantly differed compared to the control. The relative abundance of the genera Acidipropionibacterium, Enterococcus, Escherichia/Shigella, Lactobacillus, Lentilactobacillus and Propionibacterium was higher in the cheese with pink discoloration defect. The concentration of short chain fatty acids and of lactic acid in cheese was measured and a shift towards the production of propionate in the cheese with pink discoloration defect was observed. Furthermore, the possible involvement of microbially produced vitamin B12 in the formation of pink discoloration was not supported by the data, since a tendency to a lower concentration of vitamin B12 was measured in the defected cheese compared to the control.

Effect of consumption of ancient grain bread leavened with sourdough or with baker's yeast on cardio-metabolic risk parameters: a dietary intervention trial.

The aim of this study was to compare the effect of consumption of ancient grain "Verna" bread obtained by two different leavening agents, sourdough (SD) and baker's yeast (BY), on inflammatory parameters and cardiometabolic risk factors. Seventeen clinically healthy subjects were included to consume SD or BY bread for 4 weeks each, and blood analyses were carried out. The consumption of "Verna" bread obtained with both leavening agents led to a significant improvement of LDL cholesterol. A reduction of -10.6% and -8.53% was observed after replacement with SD and BY bread, respectively. A significant increase in fasting blood glucose (+6%) was observed only after the intervention with BY bread. A 10.7% decrease of vascular endothelial growth factor was found after the SD bread replacement period. The consumption of "Verna" bread resulted significantly associated with an improvement in the cardiometabolic and inflammatory profile. However, only consumption of BY bread determined a significant increase in blood glucose levels.

Isolation and characterization of indigenous Weissella confusa for in situ bacterial exopolysaccharides (EPS) production in chickpea sourdough.

Legume-based sourdough represents a potential ingredient for the manufacture of novel baked products. However, the lack of gluten of legume flours can restrict their use due to their poor technological properties. To overcome such issue, the in situ production of bacterial exopolysaccharides (EPS) during fermentation has been proposed. In this study, an EPS-producing lactic acid bacteria for in situ production in chickpea sourdough was isolated. After several backsloppings of the spontaneously fermented chickpea flour dough, a dominant strain of Weissella confusa was isolated and identified. W. confusa Ck15 was able to produce linear dextran with 2.6% α-(1 â†’ 3) linked branches, from sucrose. Temperature of 30 °C, dough yield of 333, and 2% of sucrose addition were used to produce fermented chickpea sourdoughs. The acidification and rheology of the sourdoughs inoculated with W. confusa Ck15, Leuconostoc pseudomesenteroides DSM 20193, as positive control, and Lactobacillus plantarum F8, as negative control, were compared. The in situ dextran production by W. confusa Ck15 fermentation led to the highest viscosity increase (5.90 Pa·s) and the highest EPS percentage in the doughs (1.49%), compared to the other doughs. The in situ dextran production represents a potential approach for improving the use of legume flour in bakery products; overall, this experiment represents a first step for the exploitation of microbial EPS for setting up a baking process for chickpea based product.

Technological Feature Assessment of Lactic Acid Bacteria Isolated from Cricket Powder's Spontaneous Fermentation as Potential Starters for Cricket-Wheat Bread Production.

The bacterial community profile of cricket powder highlighted the presence of four main genera: Bacteroides spp., Parabacteroides spp., Lactococcus spp., and Enterococcus spp. The spontaneous fermentation of cricket powder allowed for the isolation and characterization of seven lactic acid bacteria strains belonging to six species: Latilactobacillus curvatus, Lactiplantibacillus plantarum, Latilactobacillus sakei, Lactococcus garvieae, Weissella confusa, and Enterococcus durans. The strains were characterized and selected according to different technological properties. L. plantarum CR L1 and L. curvatus CR L13 showed the best performance in terms of general aminopeptidase activity, acidification, and growth rate in MRS broth and in dough with cricket powder and wheat flour, as well as robustness during consecutive backslopping. Thus, they were used as starter-mixed to produce sourdough to manufacture bread fortified with 20% cricket powder. The addition of cricket powder led to a significant increase of protein (up to 94%) and lipid content, from 0.7 up to 6 g/100 g of bread. Spontaneous fermentation represents a source of microbial diversity that can be exploited in order to obtain potential starters for food with innovative ingredients. Edible insects powder can be successfully added in leavened baked goods to enhance their nutritional value.

Antioxidant Properties of Sourdoughs Made with Whole Grain Flours of Hull-Less Barley or Conventional and Pigmented Wheat and by Selected Lactobacilli Strains.

The use of sourdough fermentation and whole grain flours in baked goods manufacturing are known to enhance their functional and nutritional features. In this context, it is necessary to select the most suitable lactic acid bacteria strains and flour combination to achieve this goal. A characterization of 70 lactobacilli strains based on pro-technological and nutritional properties was carried out. The screening allowed the selection of 10 strains that were used to ferment sourdoughs made with two varieties of common wheat, the conventional red-grained cv Aubusson, a blue-grained variety rich in anthocyanins cv Skorpion, and a hull-less barley variety, cv Rondo. From each fermented sourdough, a water soluble extract was obtained and evaluated for its antioxidant activity performed on cultured cells (RAW 264.7 murine macrophage) by assaying Reactive Oxygen Species (ROS) content. Sourdoughs made with pigmented wheat and barley, had an antioxidant activity greater than that recovered in those made with conventional wheat flour, in spite they have been inoculated with the same LAB strains. Results highlighted the interdependence between flour and the inoculated lactic acid bacteria that has to be taken into account for the development of healthy breads exploiting high functional value cereals through biotechnological processes.

Influence of sequential inoculum of Starmerella bacillaris and Saccharomyces cerevisiae on flavonoid composition of monovarietal Sangiovese wines.

The selection of Starmerella bacillaris strains to be used with Saccharomyces cerevisiae as mixed cultures has been recently suggested in order to produce wines containing lower ethanol and higher glycerol concentrations and to promote fructose degradation due to their fructophilic character. However, studies about effects of such mixed starter cultures on phenolic compounds, which are responsible for the colour and health-enhancing properties in red wines, are currently lacking. Therefore, in this work, the influence of sequential inoculated fermentation (SIF) with Starm. bacillaris and S. cerevisiae on phenolic content of monovarietal Sangiovese wine was evaluated by fermentations at laboratory scale. Axenic fermentations (AXFs) with S. cerevisiae were performed as control. S. cerevisiae attained higher cell densities in AXF compared with SIF. The experimental wines obtained by SIF showed significant lower ethanol and higher glycerol concentrations, whereas no significant difference was detected in colour intensity. The total phenol index reached significantly lower values in SIF. Furthermore, the wines produced by SIF contained higher concentrations of vitisin A that has a greater colour stability than the anthocyanin monomer. Finally, a lower content of both free anthocyanins and flavan-3-ols, key compounds for wine quality possessing also health-enhancing properties, was found in wines obtained by SIF. On the contrary, no significant difference was detected on flavonol concentration between SIF and AXF. This study highlighted that the use of sequential inoculum of Starm. bacillaris and S. cerevisiae can contribute to increasing the colour stability of red wines, even if at the expense of compounds with health properties.

Antioxidant and anti-inflammatory properties of sourdoughs containing selected Lactobacilli strains are retained in breads.

Sourdough fermentation influences several properties of leavened baked goods also because Lactic acid bacteria (LAB) and yeasts produce bioactive peptides with a positive effect on human health. In an early study, three Lactobacilli strains (L. farciminis H3 and A11 and L. sanfranciscensis I4) possessing different proteolytic activities were used to produce sourdoughs containing peptides equipped with anti-inflammatory and/or antioxidant properties. This work was aimed to assess whether these properties could be retained after cooking. The selected LABs were used to produce breads from which low molecular weight (LMW-) peptides were extracted. The results provide solid proofs of keeping both antioxidant and anti-inflammatory activities of peptides from cooked products. Sequences of LMW-peptides either from doughs and breads were determined by mass spectrometry: differences have been noticed in amino acidic composition and in sequences, however, all the strains produce peptides equipped with antioxidant and anti-inflammatory activities.

Development of new microalgae-based sourdough "crostini": functional effects of Arthrospira platensis (spirulina) addition.

The aim of this work was to evaluate the influence of Arthrospira platensis F&M-C256 (spirulina) incorporation on the nutritional and functional properties of "crostini", a leavened bakery product largely consumed in Italy and Europe. Sourdough was used as leavening and fermentation agent and three concentrations of A. platensis F&M-C256 were tested: 2%, 6% and 10% (w/w). Despite a lower volume increase compared to the control, the A. platensis F&M-C256 "crostini" doughs reached a technological appropriate volume after fermentation. At the end of fermentation, no significant differences in microorganisms concentrations were observed. A. platensis F&M-C256 "crostini" showed higher protein content compared to the control. Considering the European Commission Regulation on nutritional claims, "crostini" incorporated with 6% and 10% biomass can be claimed to be a "source of protein". Six and ten percent A. platensis "crostini" also presented significantly higher antioxidant capacity and phenolics. A significantly lower value of in vitro dry matter and protein digestibility between A. platensis F&M-C256 "crostini" and the control was found. The overall acceptability decreased with increasing A. platensis F&M-C256 addition. The combination of spirulina biomass addition and the sourdough technology led to the development of a novel microalgae-based bakery product with nutritional and functional features.

Extra Virgin Olive Oil Quality as Affected by Yeast Species Occurring in the Extraction Process.

In extra virgin olive oil (EVOO) extraction process, the occurrence of yeasts that could affect the quality of olive oil was demonstrated. Therefore, in this work, at first, the yeasts occurring during different extractive processes carried out in a Tuscany oil mill, at the beginning, in the middle, and the end of the harvesting in the same crop season, were quantified. Then, possible effects on quality of EVOO caused by the predominant yeast species, possessing specific enzymatic activities, were evaluated. Yeast concentrations were higher in extraction processes at the end of the harvesting. Twelve yeast species showing different isolation frequencies during olive oil extractive process and according to the harvesting date were identified by molecular methods. The yeast species dominating olive oil samples from decanter displayed enzymatic activities, potentially affecting EVOO quality according to zymogram analysis. HS-SPME-GC-MS analysis of the volatile compounds in commercial EVOO, inoculated with three yeast species (Nakazawaea molendini-olei, Nakazawaea wickerhamii, Yamadazyma terventina), pointed out significant differences depending on the strain inoculated. In conclusion, during the olive oil extractive processes, some yeast species colonize the extraction plant and may influence the chemical and sensory characteristics of EVOO depending on the cell concentrations and their enzymatic capabilities.

Use of Selected Lactobacilli to Increase γ-Aminobutyric Acid (GABA) Content in Sourdough Bread Enriched with Amaranth Flour.

γ-Aminobutyric acid (GABA) is the primary inhibitory neurotransmitter of the central nervous system and possesses various physiological functions. GABA production can be obtained thanks to lactic acid bacteria activity in different foods such as sourdoughs. Recently, breads made from blends of pseudocereals and wheat flours have attracted much attention. Amaranth is especially interesting because of its high nutritional value, having a high protein content and containing different antioxidant compounds. Therefore, this study aimed to obtain sourdough breads enriched with GABA thanks to bacterial activity and to investigate the effect of amaranth flour addition on the antioxidant and sensorial properties of bread. Eighteen lactobacilli strains were assayed for GABA production in amaranth and wheat flour liquid sourdoughs. Two strains, Lactobacillus brevis A7 and Lactobacillus farciminis A11, demonstrated high GABA producing capability; hence, they were used to prepare breads containing 20% amaranth flour. The results confirmed the capability of the two strains to increase GABA concentrations (up to 39 mg/kg) in breads. Samples with amaranth addition showed a significantly higher total phenolic content compared to the control bread (more than 15 mg GAE 100 g-1 dwb); sensory analysis showed that breads with amaranth were moderately acceptable. Nevertheless, their general liking evaluation was significantly lower compared to the control bread. The addition of pseudocereal to traditional wheat sourdough and selection of lactobacilli allowed the production of baked goods with enhanced GABA content and antioxidant capacity, but recipes have to be developed to increase the organoleptic acceptability of the final products.

Quantifying the Effects of Ethanol and Temperature on the Fitness Advantage of Predominant Saccharomyces cerevisiae Strains Occurring in Spontaneous Wine Fermentations.

Different Saccharomyces cerevisiae strains are simultaneously or in succession involved in spontaneous wine fermentations. In general, few strains occur at percentages higher than 50% of the total yeast isolates (predominant strains), while a variable number of other strains are present at percentages much lower (secondary strains). Since S. cerevisiae strains participating in alcoholic fermentations may differently affect the chemical and sensory qualities of resulting wines, it is of great importance to assess whether the predominant strains possess a "dominant character." Therefore, the aim of this study was to investigate whether the predominance of some S. cerevisiae strains results from a better adaptation capability (fitness advantage) to the main stress factors of oenological interest: ethanol and temperature. Predominant and secondary S. cerevisiae strains from different wineries were used to evaluate the individual effect of increasing ethanol concentrations (0-3-5 and 7% v/v) as well as the combined effects of different ethanol concentrations (0-3-5 and 7% v/v) at different temperature (25-30 and 35°C) on yeast growth. For all the assays, the lag phase period, the maximum specific growth rate (µmax) and the maximum cell densities were estimated. In addition, the fitness advantage between the predominant and secondary strains was calculated. The findings pointed out that all the predominant strains showed significantly higher µmax and/or lower lag phase values at all tested conditions. Hence, S. cerevisiae strains that occur at higher percentages in spontaneous alcoholic fermentations are more competitive, possibly because of their higher capability to fit the progressively changing environmental conditions in terms of ethanol concentrations and temperature.

Effect of selected strains of lactobacilli on the antioxidant and anti-inflammatory properties of sourdough.

Sourdough fermentation of cereal foods is an excellent source of obtaining peptides due to the ability of lactic acid bacteria to activate cereal proteases and produce strain-specific peptidases. With the aim of identifying the lactic acid bacterial strains potentially most effective in producing bioactive peptides, 131 lactobacilli isolates from Italian sourdoughs, used in baking technology, have been screened for proteolytic and peptidase activity. Of these, 23 strains were selected and singly inoculated in liquid sourdoughs from which a Low Molecular Weight fraction containing peptides was obtained. Evaluation of the antioxidant and anti-inflammatory activities of the extracts was performed on cultured cells (RAW 264.7 murine macrophage, murine H-end endothelium cells and Human intestinal Caco-2 cells) by assaying Reactive Oxygen Species (ROS) content, NFkB/IkB expression level and Interleukin-1ß production. As a result, three lactobacilli strains showed a high antioxidant and anti-inflammatory ability enabling the development of model sourdoughs that will potentially increase the nutritional benefits of bread.

Diversity of Saccharomyces cerevisiae Strains Isolated from Two Italian Wine-Producing Regions.

Numerous studies, based on different molecular techniques analyzing DNA polymorphism, have provided evidence that indigenous Saccharomyces cerevisiae populations display biogeographic patterns. Since the differentiated populations of S. cerevisiae seem to be responsible for the regional identity of wine, the aim of this work was to assess a possible relationship between the diversity and the geographical origin of indigenous S. cerevisiae isolates from two different Italian wine-producing regions (Tuscany and Basilicata). For this purpose, sixty-three isolates from Aglianico del Vulture grape must (main cultivar in the Basilicata region) and from Sangiovese grape must (main cultivar in the Tuscany region) were characterized genotypically, by mitochondrial DNA restriction analysis and MSP-PCR by using (GTG)5 primers, and phenotypically, by determining technological properties and metabolic compounds of oenological interest after alcoholic fermentation. All the S. cerevisiae isolates from each region were inoculated both in must obtained from Aglianico grape and in must obtained from Sangiovese grape to carry out fermentations at laboratory-scale. Numerical analysis of DNA patterns resulting from both molecular methods and principal component analysis of phenotypic data demonstrated a high diversity among the S. cerevisiae strains. Moreover, a correlation between genotypic and phenotypic groups and geographical origin of the strains was found, supporting the concept that there can be a microbial aspect to terroir. Therefore, exploring the diversity of indigenous S. cerevisiae strains can allow developing tailored strategies to select wine yeast strains better adapted to each viticultural area.

Enumeration and rapid identification of yeasts during extraction processes of extra virgin olive oil in Tuscany.

The aim of this study was to evaluate the occurrence of yeast populations during different olive oil extraction processes, carried out in three consecutive years in Tuscany (Italy), by analysing crushed pastes, kneaded pastes, oil from decanter and pomaces. The results showed yeast concentrations ranging between 10(3) and 10(5) CFU/g or per mL. Seventeen dominant yeast species were identified by random amplified polymorphic DNA with primer M13 and their identification was confirmed by restriction fragments length polymorphism of ribosomal internal transcribed spacer and sequencing rRNA genes. The isolation frequencies of each species in the collected samples pointed out that the occurrence of the various yeast species in olive oil extraction process was dependent not only on the yeasts contaminating the olives but also on the yeasts colonizing the plant for oil extraction. In fact, eleven dominant yeast species were detected from the washed olives, but only three of them were also found in oil samples at significant isolation frequency. On the contrary, the most abundant species in oil samples, Yamadazyma terventina, did not occur in washed olive samples. These findings suggest a phenomenon of contamination of the plant for oil extraction that selects some yeast species that could affect the quality of olive oil.

Typing of Lactobacillus sanfranciscensis isolates from traditional sourdoughs by combining conventional and multiplex RAPD-PCR profiles.

In the present work, a rapid and reproducible molecular method, based on the combination of conventional and multiplex RAPD-PCR reactions, was developed for typing Lactobacillus sanfranciscensis isolates from traditional sourdoughs. At first, four random primers, two used singly and two combined with the primer RD1, were chosen on the basis of their differentiating capability and reproducibility. The four resulting profiles for each isolate were integrated into a unique profile to be statistically treated by cluster analysis. The method was validated on 58 L. sanfranciscensis isolates coming from three traditional Italian sourdoughs. This new RAPD method was useful for determining the genomic diversity within the L. sanfranciscensis species. In particular, the intraspecific diversity of this species seemed to be related to the sourdough origin.