Goat milk has been described as having a better digestibility and lower allergenic properties than cow milk. The increasing demand for food safety induces consumers to prefer organic foodstuffs, which are perceived to be healthier than conventional products. The aim of this paper was to compare organic and conventional goat yoghurts marketed in Italy. The results evidenced that conventional samples showed significantly higher mean values of total solids, proteins, total lactic acid and apparent viscosity, and significantly lower mean values of pH than organic yoghurts. Moreover, the latter received significantly higher mean scores for the descriptors yoghurt flavor, while the conventional samples received significantly higher mean scores for the descriptors floury, mouth thickness and spoon thickness. The organic yoghurts differed from conventional ones for their composition and aromatic profile. The present work pointed out that conventional yoghurts are often submitted to fortification, probably with the aim of making them similar to cow yoghurts.

The aim of this study was to optimize the production of exopolysaccharides (EPS) by sourdough LactobacilluscurvatusDPPMA10 for industrial application. The effects of pH, temperature, planktonic or attached cells and of some food matrices as substrates were studied. Wheatflour hydrolysate (WFH), reconstituted skimmed milk (RSM) and whey milk were supplemented with fresh yeast extract, mineral salts, and/or molasses. Non-controlled pH, starting from 5.6 to 3.5, was the optimal condition for L. curvatusDPPMA10. Temperature of 30 °C was also found to be optimal. Solid surfaces (agar culture media) stimulated attached bacteria to synthesize EPS (≥ of two-fold, P < 0.05) with respect to planktonic cells (broth media). The highest production of EPS (ca. 46–50 g/kg of wet medium) was found during growth as attached cells in WFH agar supplemented with glucose, sucrose or molasses, mineral salts and fresh yeast extract at 30 °C for 48 h. As shown by high-performance liquid chromatography analysis, glucose was the only hydrolysis end-product for EPS synthesized during 48 h of incubation. The EPS synthesized by L. curvatusDPPMA10 improved the quality of bread and was utilized as carbon course by intestinal strains of lactobacilli and bifidobacteria. The synthesis of EPS by L. curvatusDPPMA10 under the conditions of this study may open new perspectives for their industrial applications.

Seven mature type I sourdoughs were comparatively back-slopped (80 days) at artisan bakery and laboratory levels under constant technology parameters. The cell density of presumptive lactic acid bacteria and related biochemical features were not affected by the environment of propagation. On the contrary, the number of yeasts markedly decreased from artisan bakery to laboratory propagation. During late laboratory propagation, denaturing gradient gel electrophoresis (DGGE) showed that the DNA band corresponding to Saccharomyces cerevisiae was no longer detectable in several sourdoughs. Twelve species of lactic acid bacteria were variously identified through a culture-dependent approach. All sourdoughs harbored a certain number of species and strains, which were dominant throughout time and, in several cases, varied depending on the environment of propagation. As shown by statistical permutation analysis, the lactic acid bacterium populations differed among sourdoughs propagated at artisan bakery and laboratory levels. Lactobacillus plantarum, Lactobacillus sakei, and Weissella cibaria dominated in only some sourdoughs back-slopped at artisan bakeries, and Leuconostoc citreum seemed to be more persistent under laboratory conditions. Strains of Lactobacillus sanfranciscensis were indifferently found in some sourdoughs. Together with the other stable species and strains, other lactic acid bacteria temporarily contaminated the sourdoughs and largely differed between artisan bakery and laboratory levels. The environment of propagation has an undoubted influence on the composition of sourdough yeast and lactic acid bacterium microbiotas.

This work aimed to select heat-resistant probiotic lactobacilli to be added to Fior di Latte (high-moisture cow milk Mozzarella) cheese. First, 18 probiotic strains belonging to Lactobacillus casei, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus reuteri were screened. Resistance to heating (65 or 55°C for 10 min) varied markedly between strains. Adaptation at 42°C for 10 min increased the heat resistance at 55°C for 10 min of all probiotic lactobacilli. Heat-adapted L. delbrueckii ssp. bulgaricus SP5 (decimal reduction time at 55°C of 227.4 min) and L. paracasei BGP1 (decimal reduction time at 55°C of 40.8 min) showed the highest survival under heat conditions that mimicked the stretching of the curd and were used for the manufacture of Fior di Latte cheese. Two technology options were chosen: chemical (addition of lactic acid to milk) or biological (Streptococcus thermophilus as starter culture) acidification with or without addition of probiotics. As determined by random amplified polymorphic DNA-PCR and 16S rRNA gene analyses, the cell density of L. delbrueckii ssp. bulgaricus SP5 and L. paracasei BGP1 in chemically or biologically acidified Fior di Latte cheese was approximately 8.0 log10 cfu/g. Microbiological, compositional, biochemical, and sensory analyses (panel test by 30 untrained judges) showed that the use of L. delbrueckii ssp. bulgaricus SP5 and L. paracasei BGP1 enhanced flavor formation and shelf-life of Fior di Latte cheeses.

The bacterial ecology during rye and wheat sourdough preparation was described by 16S rRNA gene pyrosequencing. Viable plate counts of presumptive lactic acid bacteria, the ratio between lactic acid bacteria and yeasts, the rate of acidification, a permutation analysis based on biochemical and microbial features, the number of operational taxonomic units (OTUs), and diversity indices all together demonstrated the maturity of the sourdoughs during 5 to 7 days of propagation. Flours were mainly contaminated by metabolically active genera (Acinetobacter, Pantoea, Pseudomonas, Comamonas, Enterobacter, Erwinia, and Sphingomonas) belonging to the phylum Proteobacteria or Bacteroidetes (genus Chryseobacterium). Their relative abundances varied with the flour. Soon after 1 day of propagation, this population was almost completely inhibited except for the Enterobacteriaceae. Although members of the phylum Firmicutes were present at very low or intermediate relative abundances in the flours, they became dominant soon after 1 day of propagation. Lactic acid bacteria were almost exclusively representative of the Firmicutes by this time. Weissella spp. were already dominant in rye flour and stably persisted, though they were later flanked by the Lactobacillus sakei group. There was a succession of species during 10 days of propagation of wheat sourdoughs. The fluctuation between dominating and subdominating populations of L. sakei group, Leuconostoc spp., Weissella spp., and Lactococcus lactis was demonstrated. Other subdominant species such as Lactobacillus plantarum were detectable throughout propagation. As shown by PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis, Saccharomyces cerevisiae dominated throughout the sourdough propagation. Notwithstanding variations due to environmental and technology determinants, the results of this study represent a clear example of how the microbial ecology evolves during sourdough preparation.

Aims:  To screen the glutamate dehydrogenase (GDH) activity of nonstarter lactic acid bacteria (NSLAB) and to determine the effects of temperature, pH and NaCl values used for cheese ripening on enzyme activity and expression of GDH gene. Methods and Results:  A subcellular fractionation protocol and specific enzyme assays were used. The effect of temperature, pH and NaCl on enzyme activity was evaluated. The expression of GDH gene was monitored by real-time PCR. One selected strain was also used as adjunct starter for cheese making to evaluate the catabolism of free amino acids and the production of volatile organic compounds (VOC) during cheese ripening. The cytoplasm fraction of all strains showed in vitro NADP-dependent GDH activity. NADP-GDH activity was markedly strain dependent and varied according to the interactions between temperature, pH and NaCl. Lactobacillus plantarum DPPMA49 showed the highest NADP-GDH activity under temperature, pH and NaCl values found during cheese ripening. RT-PCR analysis revealed that GDH expression of Lact. plantarum DPPMA49 was down-expressed by low temperature (&lt;13°C) and over-expressed by NaCl (1·87–5·62%). According to NADP-GDH activity, the highest level of VOC (alcohols, aldehydes, miscellaneous and carboxylic acids) was found in cheeses made with DPPMA49. Conclusions:  The results of this study may be considered as an example of the influence of temperature, pH and NaCl on enzyme activity and expression of functional genes, such as GDH, in cheese-related bacteria. Significance and Impact of the Study:  It focuses on the phenotypic and molecular characterization of the NADP-GDH in lactobacilli under cheese-ripening conditions. The findings of this study contribute to the knowledge about enzymes involved in the catabolism of amino acids, to be used as an important selection trait for cheese strains.

This work showed the effect of pheromone plantaricinA (PlnA) on the proliferation and migration of the humankeratinocytes NCTC 2544. PlnA was chemically synthesized and used as pure peptide or biologically synthesized during co-cultivation of Lactobacillusplantarum DC400 and Lactobacillus sanfranciscensis DPPMA174. The cell-free supernatant (CFS) was used as the crude preparation containing PlnA. The inductive effect of PlnA on the proliferation of NCTC 2544 cells was higher than that found for hyaluronic acid, a well known skin protective compound. As shown by scratch assay and image analyses, PlnA enhanced the migration of NCTC 2544 cells. Compared to the basal serum free medium (control), the highest inductive effect was found using 10 μg/ml of chemically synthesized PlnA. Similar results (P &gt; 0.05) were found for CFS. In agreement, the percentage of the starting scratch area was decreased after treatment (24 h) with PlnA. The expression of transforming growth factor-β1 (TGF-β1), keratinocyte growth factor 7 (FGF7), vascular endothelial growth factor (VEGF-A), and interleukin-8 (IL-8) genes was affected by PlnA. Compared to control, TGF-β1gene was under expressed in the first 4 h of treatments and up-regulated after 8–24 h. On the contrary, FGF7gene was strongly up-regulated in the first 4 h of treatments. Compared to control, VEGF-A and IL-8genes were always up-regulated during the 4–24 h from scratching. Since capable of promoting the proliferation and migration of the humankeratinocytes and of stimulating IL-8 cytokine, the use of PlnA for dermatological purposes should be considered.

This study aimed at investigating the robustness of selected sourdough strains of Lactobacillusplantarum. Seven strains were singly used as sourdoughtype I starters under daily back-slopping propagation (ten days) using wheatflour. Cell numbers of presumptive lactic acid bacteria varied slightly (median values of 9.13–9.46 log cfu g−1) between and within started sourdoughs, as well as the acidifying activity (median values of 1.24–1.33). After three days also the control sourdough (unstarted) had the same values. As shown by RAPD-PCR analysis, five (DB200, 3DM, G10C3, 12H1 and LP20) out of seven strains maintained elevated cell numbers (ca. 9 log cfu g−1) throughout ten days. The other two strains progressively decreased to less than 5 log cfu g−1. As identified by partial sequencing of 16S rRNA and recA genes, L. plantarum (11 isolates), pediococci (7), Lactobacillus casei (3) and Lactobacillus rossiae (2) dominated the flour microbiota. Monitoring of lactic acid bacteria during sourdoughpropagation was carried out by culture dependent approach and using PCR-DGGE (Denaturing Gradient Gel Electrophoresis). Except for the sourdough started with L. plantarum LP20, in all other sourdoughs at least one autochthonous strain of L. plantarum emerged. All emerging strains of L. plantarum showed different RAPD-PCR profiles. L. rossiae and Pediococcus pentosaceus were only found in the control and sourdough started with strain 12H1. The characterization of the catabolic profiles of sourdoughs (Biolog System) showed that sourdoughs containing persistent starters behaved similarly and their profiles were clearly differentiated from the others. One persistent strain (DB200) of L. plantarum and Lactobacillus sanfranciscensis LS44, previously shown to be persistent (Siragusa et al., 2009), were used as the mixed starter to produce a wheatfloursourdough. Both strains cohabited and dominated during ten days of propagation.

The use of sourdough, even in combination with cryoprotectant (skim milk, sucrose and trehalose), conventional additives (guar gum, diacetyl tartaric acid esters of monoglycerides, ascorbic acid), honey or fructose and glucose, in frozen dough technology was investigated. After frozen storage, the leavening performance of doughs, and the hardness and texture of breads were compared to those of an unfrozen dough, and to those of a conventional frozen dough. All frozen doughs showed a longer fermentation time and a lower volume increase, with respect to unfrozen dough. When sourdough was combined with cryoprotectant, honey or both, the leavening performance improved compared to the use of sourdough alone. Compared to the conventional frozen dough, higher leavening performance was reached combining sourdough with cryoprotectant alone or together with honey. Sourdough combined with honey, fructose and glucose, honey and cryoprotectant, or conventional additives decreased bread hardness compared to the unfrozen dough bread and to the conventional frozen dough bread. Independently from the use of sourdough, conventional additives allowed to reach a specific volume not significantly different from that of unfrozen dough bread, and breads containing honey were characterized by low values of hardness and by high values of red index.

Bacteria belonging to the genus Enterococcus spp.—a member of the lactic acid bacteria group—are natural inhabitants of various environments, including vegetable products. Although some strains show pathogenic determinants, overall these bacteria may have some protechnological features. Some enterococci have been described as potential starter or protective cultures in the dairy industry, since they contribute to the organoleptic and quality characteristics of dairy products. Although several fermented vegetable products have a long history in human nutrition, studies regarding autochthonous enterococci and their application to fermented vegetable foods are much less numerous than those concerning dairy foods. In this review, after a general overview of enterococci, their presence and role in fermented vegetable foods (including table olives, sauerkraut, kimchi, tomato juice, French beans, caper berries and cereal-based products) will be covered.

Plantaricin A (PlnA) is a peptide with antimicrobial and pheromone activities. PlnA was synthesized chemically and used as a pure peptide or synthesized biologically using Lactobacillus plantarum DC400 co-cultured with Lactobacillus sanfranciscensis DPPMA174. Cell-free supernatant (CFS) was used as a crude PlnA preparation. As estimated using the 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide and the 2',7'-dichlorofluorescein diacetate assays, both PlnA preparations increased the antioxidant defenses of human NCTC 2544 keratinocytes. PlnA (10 ?g/ml) had a higher activity than hyaluronic acid or 125 ?g/ml ?-tocopherol. Effects on the transcriptional regulation of filaggrin (FLG), involucrin (IVL), hyaluronan synthase (HAS2), human ?-defensin-2 (HBD-2) and tumor necrosis factor-alpha (TNF-?) genes were assayed. Compared with the control, expression of the FLG gene in NCTC 2544 cells increased in cells treated with hyaluronic acid, 1 or 10 ?g/ml PlnA. Compared with the control, the level of IVL gene expression increased in NCTC 2544 cells treated with 10 ?g/ml PlnA. No significant difference was found between the level of the HAS2 gene expressed by control cells and cells treated with PlnA. Compared with chemically synthesized PlnA, the up-regulation of the HBD-2 gene by CFS was higher. Compared with the control, expression of TNF-? decreased in NCTC 2544 cells after treatment with 1 or 10 ?g/ml of chemically synthesized PlnA. In contrast, the level of TNF-? was highest in the presence of 10 ?g/ml CFS-PlnA. These findings suggest that the PlnA was positively sensed by human keratinocytes, promoting antioxidant defenses, barrier functions and antimicrobial activity of the skin.

Gut microbiota, the largest symbiont community hosted in human organism, is emerging as a pivotal player in the relationship between dietary habits and health. Oral and, especially, intestinal microbes metabolize dietary components, affecting human health by producing harmful or beneficial metabolites, which are involved in the incidence and progression of several intestinal related and non-related diseases. Habitual diet (Western, Agrarian and Mediterranean omnivore diets, vegetarian, vegan and gluten-free diets) drives the composition of the gut microbiota and metabolome. Within the dietary components, polymers (mainly fibers, proteins, fat and polyphenols) that are not hydrolyzed by human enzymes seem to be the main leads of the metabolic pathways of gut microbiota, which in turn directly influences the human metabolome. Specific relationships between diet and microbes, microbes and metabolites, microbes and immune functions and microbes and/or their metabolites and some human diseases are being established. Dietary treatments with fibers are the most effective to benefit the metabolome profile, by improving the synthesis of short chain fatty acids and decreasing the level of molecules, such as p-cresyl sulfate, indoxyl sulfate and trimethylamine N-oxide, involved in disease state. Based on the axis diet-microbiota-health, this review aims at describing the most recent knowledge oriented towards a profitable use of diet to provide benefits to human health, both directly and indirectly, through the activity of gut microbiota.

Food allergy is recognized as one of the major health concerns. It is estimated that ca. 4% of the population is affected by food allergenic disorders. Food allergies are defined as IgE-mediated hypersensitivity reactions. Foods such as peanuts, tree nuts, wheat, soy, cow's milk, egg, fish and shellfish are regarded as responsible for the majority of reactions. The ubiquitous presence of allergens in the human foods coupled with an increased awareness of food allergies warrants to undertake appropriate preventive measures for protecting sensitive consumers from unwanted exposure to offending food allergens. 2-DE followed by immunoblotting and identification of IgE-reactive proteins, as a proteomic approach to identify new allergens in foods, are reviewed. Specific examples of identification of allergens in foods and beverages by using 2-DE and IgE are described. Protein profiling using 2-DE and allergens detection by IgE has become a powerful method for analyzing changes of allergens content in complex matrix during food processing.

The aim of this study was to assess whether wheat endophytic lactic acid bacteria (LAB) are able to dominate in sourdough ecosystem. To do that, a first experimental phase considered doughs produced under semi-sterile conditions and singly inoculated with different strains of endophytic LAB and Lactobacillus sanfranciscensis A4 isolated from sourdough. Notwithstanding the high frequency of Lactobacillus plantarum in the sourdoughs prepared in laboratory, only one of the starter strains, L. plantarum LB2, was detected after five days of back-slopping. Subsequently, the ability of this strain to dominate traditional sourdoughs was evaluated at bakery and laboratory level. Contamination of sourdoughs with L. plantarum LB2 caused an increased number of LAB and, accordingly, higher acidification, compared to the sourdoughs before this event. After six days of propagation, the wheat endophytic strain L. plantarum LB2 was retrieved as a component of the bacterial population, in all the sourdoughs and regardless of the place of propagation. In addition, the contamination event caused a modification of the lactic acid bacterium biota, which in turn influenced some sourdoughs biochemical features. In conclusion, this study showed that wheat endophytic LAB could represent a potential reservoir for selecting robust strains to be used as sourdough starters.