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Global concerns over folate deficiency, the risks of excessive synthetic folic acid consumption, and food loss implications for environmental sustainability and food security drive needs of innovative approaches that align food by-product valorisation with folate bio-enrichment. This study explored the use of three fruit by-products extracts (grape, passion fruit, and pitaya) and whey to develop a folate bio-enriched fermented whey-based beverage. Three strains (Lacticaseibacillus rhamnosus LGG, Bifidobacterium infantis BB-02, and Streptococcus thermophilus TH-4) were tested for folate production in different fermentation conditions in modified MRS medium and in a whey-based matrix prepared with water extracts of these fruit by-products. B. infantis BB-02 and S. thermophilus TH-4, alone and in co-culture, were the best folate producers. The selection of cultivation conditions, including the presence of different substrates and pH, with grape by-product water extract demonstrating the most substantial effect on folate production among the tested extracts, was crucial for successfully producing a biofortified fermented whey-based beverage (FWBB). The resulting FWBB provided 40.7 μg of folate per 100 mL after 24 h of fermentation at 37 °C, effectively leveraging food by-products. Moreover, the beverage showed no cytotoxicity in mouse fibroblast cells tests. This study highlights the potential for valorising fruit by-products and whey for the design of novel bioenriched foods, promoting health benefits and contributing to reduced environmental impact from improper disposal., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Lactic acid bacteria are probiotics in the intestines and have been widely used as natural antioxidants in the food industry. In this study, Enterococcus italicus FM5 with strong antioxidant ability was isolated from fresh milk. The safety evaluation showed that E. italicus FM5 was sensitive to ampicillin, chloramphenicol, erythromycin, vancomycin, rifampicin, and tetracycline, and was not hemolytic. Meanwhile, the whole genome information and biofunctional attributes of this strain were determined and analyzed. Subsequently, E. italicus FM5 was co-cultured with traditional yogurt starters (Streptococcus thermophilus and Lactobacillus bulgaricus) to make fermented milk. The results showed that the addition of E. italicus FM5 could improve the oxygen free radical scavenging ability of the fermented milk, and the scavenging rates of DPPH, ABTS, OH - , and O 2 - radicals reaching up to 95.54 %, 88.35 %, 93.65 %, and 60.29 %, respectively. Furthermore, the addition of E. italicus FM5 reduced the curd time and improved the water holding capacity of the fermented milk. Besides, the growth of Lb. bulgaricus was significantly promoted when co-cultured with E. italicus FM5, thus the survival cells were increased compared with the traditional fermentation processes. Therefore, this study emphasized the potential to manufacture fermented milk by the co-cultivation of E. italicus with traditional yogurt starters., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

2'-Fucosyllactose (2'-FL) is postulated to provide health benefits and promote the growth of probiotics. This work was undertaken to study the effects of 2'-FL on the viability of starter cultures and Bifidobacterium strains of human origin in yogurt during refrigerated storage. Yogurts were produced containing 2'-FL (0 or 2 g/L) and Bifidobacterium strains of human origin (Bifidobacterium longum subsp. longum BB536 or Bifidobacterium longum subsp. infantis ATCC 15697) at a concentration of at least 10 9 CFU/mL. All yogurts were stored at 4°C for 5 weeks. Results showed that 2'-FL was stable in yogurts for at least 5 weeks of cold storage, and the addition of 2'-FL did not significantly alter yogurt fermentation parameters, associated metabolites, and the viability of mixed yogurt starter cultures and Bifidobacterium strains (p > 0.05). The addition of bifidobacteria had a negative impact (p < 0.05) on the survival rate of starter cultures, Streptococcus thermophilus and Lactobacillus delbureckii subsp. bulgaricus. Meanwhile, it is difficult to maintain a high survival rate of bifidobacteria in final yogurt products, and the addition of 2'-FL could not enhance the viability of bifidobacteria. B. longum BB536 survived at a level higher than 10 6 CFU/g for 28 days, while B. infantis ATCC15697 maintained this level for only 7 days. In summary, this study has shown the impact of 2'-FL and bifidobacterial species on yogurt properties, and results suggest that it is promising to use 2'-FL in yogurt products as a prebiotic. PRACTICAL APPLICATION: Yogurt is known for its beneficial effects on human health and nutrition. This study reported the production of symbiotic yogurt containing bifidobacteria and 2'-fucosyllactose (2'-FL) as a functional food for specified health uses. The viability of yogurt starter cultures and probiotic bifidobacterial strains was analyzed in this study. Moreover, this research demonstrated that 2'-FL could be added to yogurt without affecting the characteristics of yogurt significantly., (© 2024 Institute of Food Technologists.)

This study investigated the bioconversion and bioaccessibility of soy isoflavones produced in sogurt fermented with S. thermophilus and L. bulgaricus during in vitro digestion. The highest survivability of S. thermophilus (6.49 log cfu/mL) and L. bulgaricus (6.48 log cfu/mL) was in oral phase. In gastric phase, the total aglycones of sogurt (26.73 g/L) increased up to 20 times than control (1.21 g/L), with a significant increase in daidzein (17.05 g/L) and genistein (9.68 g/L). Addition of 8U of β-glucosidase into soymilk significantly increased the conversion of isoflavone in ENTII (daidzein: 0.46 g/L; genistein: 0.18 g/L) than in ENTI (daidzein: 0.33 g/L; genistein: 0.20 g/L). The particle size analysis and confocal micrographs of digesta also suggest the size of fat and protein in gastric phase to be smaller than in intestinal phase. The results indicate the prospective to develop soy-based fermented products capable of releasing high isoflavone in the digestive system., (Copyright © 2020. Published by Elsevier Ltd.)

Enumeration and isolation of Streptococcus salivarius subsp. thermophilus from cheese is challenging, due to the relatively high number of species it may host. We describe medium SPY9.3 for the cultivation of S. salivarius subsp. thermophilus from cheese. The medium and related incubation conditions (SPY) was compared with 2 other protocols, M17 and ST: sensitivity was assessed by parallel cultivation of 55 strains of S. salivarius subsp. thermophilus, and selectivity by (i) parallel cultivation of 60 strains belonging to 20 different non-target species and sub-species and (ii) isolating bacteria from 3 raw-milk cheeses. Colony counts were similar on SPY9.3 and M17 (mean difference 0.07 log(cfu/mL), p > 0.001) and significantly higher on ST than on M17 and SPY9.3 (mean differences 0.42 and 0.48 log(cfu/mL), respectively, p < 0.001). SPY was more specific than ST and M17, with respectively 20%, 40%, and 50% of the investigated non-target species able to grow. S. salivarius subsp. thermophilus, Enterococcus spp., and Staphylococcus aureus were indistinguishable using all 3 protocols. Only SPY avoided growth of Lactobacillus delbrueckii subsp. lactis. Finally, ST and SPY displayed higher recoveries of S. salivarius subsp. thermophilus colonies from cheese than M17 (5.6, 5.5, and 3.0 adjusted log(cfu/mL), respectively) and the lowest proportion of non-specific isolates. The protocol described here and based on SPY9.3 presents a promising alternative to existing protocols for the enumeration and isolation of S salivarius subsp. thermophilus from cheese or other complex fermented products., (Copyright © 2020 Agroscope. Published by Elsevier Ltd.. All rights reserved.)

Lactic acid bacteria fermentation is a safe and green technology that can modify the function of food ingredients (including proteins). In this article, egg yolks were subjected to fermentation with commercial lactic acid bacteria for 0, 3, 6 and 9 h, respectively. After fermentation treatment, the microbial composition has changed obviously (Streptococcus thermophilus increased significantly). The free sulfhydryl group (SH) contents and surface hydrophobicity of egg yolk proteins were significantly reduced. The rheological results indicated that the treated egg yolks possessed a decreased apparent viscosity. Correspondingly, the emulsifying activity of egg yolk was enhanced from 9.07 to 19.55, 23.40 and 24.61 m 2 /g for 3, 6 and 9 h of fermentation, respectively. And the emulsifying stability reached the maximum after 3 h of fermentation. This study investigated the relationship between structure and properties of yolk proteins, and showed that lactic acid fermentation endued egg yolk with better emulsifying properties., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Camel milk, similar to cow milk, contains all of the essential nutrients as well as potentially health-beneficial compounds with anticarcinogenic, antihypertensive, and antioxidant properties. Camel milk has been used for the treatment of allergies to cow milk, diabetes, and autism. Camel milk helps decrease cholesterol levels in blood and improves metabolism. One of the most desirable food tastes is sweetness. However, the excessive ingestion of sugar negatively affects human health. Monk fruit sweetener is a natural, 0-calorie sweetener with many health-beneficial functions. Monk fruit sweetener helps decrease symptoms of asthma and diabetes, prevents oxidation and cancer, protects the liver, regulates immune function, and lowers glucose levels. Monk fruit sweetener is 100 to 250 times sweeter than sucrose. The objective of this study was to examine the influence of different concentrations of monk fruit sweetener on the physicochemical properties and microbiological counts of drinking yogurt made from camel milk. Camel milk drinking yogurt was produced with 0, 0.42, 1.27, and 2.54 g/L of monk fruit sweetener and stored for 42 d. The physicochemical characteristics and microbiological counts of yogurts were measured at d 1, 7, 14, 21, 28, 35, and 42. For the physicochemical characteristics, pH, titratable acidity, viscosity, and color [lightness-darkness (L*), red-green axis (a*), yellow-blue axis (b*), chroma (C*), and hue angle (h*)] values were evaluated. The counts of Streptococcus thermophilus, Lactobacillus bulgaricus, Lactobacillus acidophilus, coliforms, and yeast and mold were determined. Three replications were conducted. The sweetener addition significantly influenced pH, viscosity, and color (a*, b*, C*, and h*) values. Control samples had significantly higher pH values, lower viscosity, lower b* and C* values, and higher h* values than the samples with 1.27 and 2.54 g/L of monk fruit sweetener. Growth of S. thermophilus, L. bulgaricus, and probiotic culture L. acidophilus was not affected by the incorporation of monk fruit sweetener. Monk fruit sweetener can be added in camel milk yogurts as a health-beneficial 0-calorie sweetener., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Societal demand for plant-based foods is increasing. In this context, soya products fermented using lactic acid bacteria (LAB) are appealing because of their potential health and nutritional benefits. The thermophilic LAB Streptococcus thermophilus is an essential starter species in the dairy industry. However, while its physiology is well characterized, little is known about its general metabolic activity or its techno-functional properties when it is grown in soya milk. In this study, S. thermophilus LMD-9 growth, sugar production, and lactic acid production in soya milk versus cow's milk were measured. Additionally, the main metabolic pathways used by the bacterium when growing in soya milk were characterized using a proteomic approach. Streptococcus thermophilus LMD-9 growth decreased soya milk pH, from 7.5 to 4.9, in 5 h. During fermentation, acidification thus occurred in tandem with lactate production and increasing population size (final population: 1.0 × 10 9  CFU/ml). As growth proceeded, sucrose was consumed, and fructose was produced. The proteomic analysis (LC-MS/MS) of the strain's cytosolic and cell envelope-associated proteins revealed that proteins related to amino acid transport and nitrogen metabolism were the most common among the 328 proteins identified (63/328 = 19.2% of total proteins). The cell-wall protease PrtS was present, and an LMD-9 deletion mutant was constructed by interrupting the prtS gene (STER&#95;RS04165 locus). Acidification levels, growth levels, and final population size were lower in the soya milk cultures when the ΔprtS strain versus the wild-type (wt) strain was used. The SDS-PAGE profile of the soluble proteins in the supernatant indicated that soya milk proteins were less hydrolyzed by the ΔprtS strain than by the wt strain. It was discovered that S. thermophilus can grow in soya milk by consuming sucrose, can hydrolyze soya proteins, and can produce acidification levels comparable to those in cow's milk. This study comprehensively examined the proteomics of S. thermophilus grown in soya milk and demonstrated that the cell-wall protease PrtS is involved in the LAB's growth in soya milk and in the proteolysis of soya proteins, which are two novel findings. These results clarify how S. thermophilus adapts to soya milk and can help inform efforts to develop new fermented plant-based foods with better-characterized biochemical and microbiological traits., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A plain symbiotic almond yogurt-like product was formulated and developed using a plant-based starter YF-L02 (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus supplemented with Lactobacillus acidophilus, Lactobacillus paracasei, and Bifidobacterium animalis) and inulin; 0.6% polymerized whey protein (PWP), 0.3% pectin, and 0.05% xanthan gum were optimized for the formula of the almond yogurt alternative. Two groups with/without calcium citrate and vitamin D 2 were prepared and analyzed for chemical composition, changes in pH, viscosity, and probiotic survivability during storage at 4 °C for 10 weeks. The results showed that (1) over 10 weeks storage, the differences in the pH, viscosity, and probiotic survivability between the control and the fortified samples were not significant (P > 0.05); (2) the pH of both yogurt samples decreased 0.2 units while their viscosity slightly increased during storage; (3) the populations of L. paracasei and B. animalis remained above 10 6 cfu/g during the storage, whereas the population of L. acidophilus decreased dramatically during the first 4 weeks, especially the control group; (4) the microstructure was examined by scanning electron microscopy, revealing a compact and denser gel structure formed by 0.6% PWP with the presence of 0.3% pectin and 0.05% xanthan gum. In conclusion, PWP might be a proper gelation agent for the formulation of symbiotic almond yogurt alternative. PRACTICAL APPLICATION: In this study, polymerized whey protein was used as a gelation agent to formulate symbiotic almond yogurt alternatives with comparable physical texture and probiotic survivability to dairy yogurt during storage. This technology may be used for the development of plant-based fermented foods., (© 2020 Institute of Food Technologists®.)

In streptococci, intracellular quorum sensing pathways are based on quorum-sensing systems that are responsible for peptide secretion, maturation, and reimport. These peptides then interact with Rgg or ComR transcriptional regulators in the Rap, Rgg, NprR, PlcR, and PrgX (RRNPP) family, whose members are found in Gram-positive bacteria. Short hydrophobic peptides (SHP) interact with Rgg whereas ComS peptides interact with ComR regulators. To date, in Streptococcus thermophilus , peptide secretion, maturation, and extracellular fate have received little attention, even though this species has several (at least five) genes encoding Rgg regulators and one encoding a ComR regulator. We studied pheromone export in this species, focusing our attention on PptAB, which is an exporter of signaling peptides previously identified in Enterococcus faecalis , pathogenic streptococci and Staphylococcus aureus . In the S. thermophilus strain LMD-9, we showed that PptAB controlled three regulation systems, two SHP/Rgg systems (SHP/Rgg 1358 and SHP/Rgg 1299 ), and the ComS/ComR system, while using transcriptional fusions and that PptAB helped to produce and export at least three different mature SHPs (SHP 1358 , SHP 1299 , and SHP 279 ) peptides while using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using a deep sequencing approach (RNAseq), we showed that the exporter PptAB, the membrane protease Eep, and the oligopeptide importer Ami controlled the transcription of the genes that were located downstream from the five non-truncated rgg genes as well as few distal genes. This led us to propose that the five non-truncated shp / rgg loci were functional. Only three shp genes were expressed in our experimental condition. Thus, this transcriptome analysis also highlighted the complex interconnected network that exists between SHP/Rgg systems, where a few homologous signaling peptides likely interact with different regulators.

Background: This study aimed to evaluate the protective effect of different biopolymer systems on the viability of two probiotics (Lactobacillus rhamnosus and Streptococcus thermophilus) during storage and in vitro digestion. Methylcellulose (MC), sodium alginate (SA), and whey protein (WP)-based structures were designed and characterized in terms of pH, rheological properties, and visual appearance., Results: The results highlighted that the WP-system ensured probiotic protection during both storage and in vitro digestion. This result was attributed to a combined effect of the physical barrier offered by the protein gel network and whey proteins as a nutrient for microbes. On the other hand, surprisingly, the viscous methylcellulose-based system was able to guarantee good microbial viability during storage. However, this was not confirmed during in vitro digestion. The opposite results were obtained for sodium alginate beads., Conclusion: The results suggest that the capacity of a polymeric structure to protect probiotic bacteria is a combination of structural organization and system formulation. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

β-galactosidases, commonly referred to as lactases, are used for producing lactose-free dairy products. Lactases are usually purified from microbial sources, which is a costly process. Here, we explored the potential that lies in using whole cells of a food-grade dairy lactic acid bacterium, Streptococcus thermophilus, as a substitute for purified lactase. We found that S. thermophilus cells, when treated with the antimicrobial peptide nisin, were able to hydrolyze lactose efficiently. The rate of hydrolysis increased with temperature; however, above 50 °C, stability was compromised. Different S. thermophilus strains were tested, and the best candidate was able to hydrolyze 80% of the lactose in a 50 g/L solution in 4 h at 50 °C, using only 0.1 g/L cells (dry weight basis). We demonstrated that it was possible to grow the cell catalyst on dairy waste, and furthermore, that a cell-free supernatant of a culture of a nisin-producing Lactococcus lactis strain could be used instead of purified nisin, which reduced cost of use significantly. Finally, we tested the cell catalysts in milk, where lactose also was efficiently hydrolyzed. The method presented is natural and low-cost, and allows for production of clean-label and lactose-free dairy products without using commercial enzymes from recombinant microorganisms. KEY POINTS: • Nisin-permeabilized Streptococcus thermophilus cells can hydrolyze lactose efficiently. • A low-cost and more sustainable alternative to purified lactase enzymes. • Reduction of overall sugar content. • Clean-label production of lactose-free dairy products.

In this study, we modified reinforced clostridial medium (RCM) to selectively enumerate and isolate Lactobacillus delbrueckii ssp. bulgaricus, a probiotic and important starter culture in the dairy industry. The disparity in the reported carbohydrate fermentation pattern of L. delbrueckii ssp. bulgaricus was used to develop a growth medium not only selective for L. delbrueckii ssp. bulgaricus but significantly inhibitory to the growth of other lactic acid bacteria. A recently modified RCM (mRCM) was optimized for this study by the addition of 0.5% fructose, 0.5% dextrose, 1% maltose, and 0.25% sodium pyruvate while replacing lactose as a carbohydrate source. The cell recovery and bacterial counts of L. delbrueckii ssp. bulgaricus in tested products (pure L. delbrueckii ssp. bulgaricus strains, starter culture, probiotic supplements, and yogurt) using our mRCM with sodium pyruvate (mRCM-PYR) were significantly higher than in the recently modified RCM and the common de Man, Rogosa, and Sharpe (MRS) culture medium. The growth of other lactic acid bacteria (Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus reuteri) and Bifidobacteria was retarded in this modified medium compared with their growth in MRS and mRCM. This result is a significant improvement in the enumeration and differentiation of L. delbrueckii ssp. bulgaricus in mRCM-PYR compared with the results in MRS and mRCM where the high background growth of similar species interferes with the accuracy of bacterial population counts. Our results thus suggest that mRCM-PYR could be recommended as a reliable alternative growth medium for the selective enumeration and isolation of L. delbrueckii ssp. bulgaricus in a mixed culture., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In the dairy industry, exopolysaccharides (EPS) produced in situ from lactic acid bacteria are of great interest because of their contribution to product texture. Some EPS cause ropiness which might be linked to specific physical and chemical EPS properties. EPS show a broad variety of chemical structures and, because analysis is rather complex, it is still a major challenge to establish structure-function relationships. The aim of this study was to produce EPS with different degree of ropiness, perform in-depth structural elucidations and relate this information to their behaviour in aqueous solutions. After cultivation of Streptococcus thermophilus DGCC7919 and Lactococcus lactis LL-2A and subsequent EPS isolation, both EPS showed similar macromolecular properties, but pronounced differences in monosaccharide composition and glycosidic linkages. Our data suggests that mainly the side chains in the EPS from LL-2A might be responsible for a higher ropiness than that observed for EPS from DGCC7919., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Thermophilic and mesophilic lactic acid bacteria (LAB), such as Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, and Lactococcus lactis, play a crucial role in the technological and sensory quality of Mozzarella cheese. In this study, the safety (genes encoding virulence factors and antibiotic resistance) and acidifying activity of autochthonous S. thermophilus cultures were evaluated in order to choose the most suitable strain for industrial application. The safe and good acidifying culture was tested in two buffalo Mozzarella cheese batches: Mozzarella cheeses produced with autochthonous culture (SJRP107) and commercial culture (STM5). The cultivable LAB was evaluated by culture-dependent method (plate counting) and the quantification of S. thermophilus cultures (commercial and autochthonous) were evaluated by culture-independent method RealT-qPCR (real-time quantitative polymerase chain reaction). The texture, physicochemical and proteolytic properties of the Mozzarella cheeses were similar for both batches. The nonstarter LAB count was higher during manufacture than in the storage, and the RealT-qPCR indicated the presence of S. thermophilus culture until the end of storage. S. thermophilus SJRP107 presented high potential for safety application in the production of Mozzarella cheese. Furthermore, considering the culture characteristics and their relationship with product quality, further studies could be helpful to determine their effect on the sensory characteristics of the cheese., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Streptococcus thermophilus ASCC 1275 has two chain length determining genes - epsC and epsD- in its eps gene cluster, and produces two times more EPS in sucrose medium than that in glucose and lactose. Hence, we investigated the influence of sugars (glucose, sucrose and lactose), at log phase (5 h) and stationary phase (10 h), on the global proteomics of S. thermophilus 1275 to understand the differentially expressed proteins (DEPs) during EPS production using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis. Among 98 DEPs in sucrose medium, most of them were mapped into EPS biosynthesis pathway and other related metabolisms. There was an upregulation of several proteins involved in sugar transport (phosphoenolpyruvate (PEP) phosphotransferase system), EPS assembly (epsG1D) and amino acid metabolism (methionine, cysteine/arginine metabolism) in sucrose medium. This study showed that increased EPS production in S. thermophilus 1275 requires a well-co-ordinated regulation of pathway involved in both EPS assembly and amino acid metabolism along with the availability of sugars. Thus, it provided valuable insights into the biosynthesis and regulation of EPS in S. thermophilus 1275, and potential gene targets for understanding high-EPS strains.

This work was carried out to pursue a double objective: to improve the hygienic safety of cheeses produced from raw ewes' milk; and to produce a new typology of raw ewes' milk through the application of "Grana" technology for which the name "Gran Ovino" was chosen. With this in mind, raw milk from an individual farm was transformed under controlled conditions at a dairy pilot plant. The production technology included the partial skimming of the evening and morning milk mixture by cream surfacing and the addition of a natural whey starter cultures (NWSC) prepared with four selected Streptococcus thermophilus strains (PON6, PON244, PON261 e PON413). Ten microbial groups were investigated by plate counts from raw milk until ripened cheeses. Lactic acid bacteria (LAB) were in the range 10 4 -10 5  CFU/ml before NWSC addition. After curdling, this group increased by 3 log cycles and was counted at 10 6  CFU/g after curd cooking. A rapid pH drop (to 6.05) was registered after almost 3 h from NWSC addition. The levels of members of the Enterobacteriaceae family were at about 10 3  CFU/ml in raw milk and decreased after curd cooking to 1 log cycle. A similar behavior was shown by the other undesired microbial groups and a complete disappearance of staphylococci was registered. The microbiological counts of 9-month ripened cheeses showed the dominance of LAB and undetectable levels of the undesired bacteria. MiSeq Illumina was applied to better investigate the bacterial composition of ripened cheeses and this technique evidenced that the majority of OTUs belonged to Lactobacillus and Streptococcus genera. The final cheeses were characterized by 67.65% dry matter of which 41.85% of fats and 47.02% of proteins. The main cheese fatty acids were palmitic, oleic and myristic acids and the saturated fatty acids/unsaturated fatty acids ratio was 2.17. Forty-one volatile compounds, including acids, esters, ketones, alcohols, aldehydes, phenols and one terpene were emitted from the cheese. Sensory evaluation showed a general appreciation for the new cheese product by judges., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Understanding global changes of physiological processes at the molecular level during the growth of Streptococcus thermophilus is essential for the rational design of cultivation media and the optimization of bioprocesses. Transcriptomics and proteomics were combined to investigate the global changes at the transcript and protein level during the growth of S. thermophilus. The expression of 1396 genes (FDR ≤ 0.001) and 876 proteins (P < 0.05) changed significantly over time. The most remarkable growth phase dependent changes occurred in the late-lag phase and were related to heterofermentation, glycolysis, peptidoglycan biosynthesis, conversion between amino acids and stress response. The present results could provide theoretical guidance for high-cell-density culture, help design cultivation media, and help attain a high biomass of S. thermophilus.

Preparation of selenium-enriched probiotics and Se-enrichment of probiotic-fermented blended juices were performed and optimized using orthogonal test. Se content had a significant 13.0-fold increase by the addition of 1% Se-enriched Streptococcus thermophilus starters in juice fermentation. Chemical properties of fermented blended juices were determined. Reducing sugar content decreased significantly after fermentation, and the same downtrend was observed for free amino acids and organic acids, with lactic acid being an exception. Meanwhile, dynamic variation analysis of flavor components during the fermentation, and characteristic aroma-active compounds before and after fermentation were demonstrated by GC-MS and GC-O. Eleven aroma-active substances were identified from juices without fermentation, while 7 characteristic compounds were detected in fermented juices. Furthermore, potential correlations between chemical and flavor characteristics were explored based on multivariate statistical analysis. These results indicate that a potential Se-enriched fermented beverage was established, and the fermentation process led to differences in the chemical substrates and impact odorants., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Recently, interest in the development of functional foods enriched with bioactive components has increased. Dairy products supplemented with tea extracts known for their health-promoting properties are good examples of such products. However, most of the scientific studies and applications focus on green tea. The present study was established to estimate the effect of Pu-erh tea supplementation on the viability of starter microorganisms and selected physico-chemical and sensory properties of probiotic ABT-yoghurt., Methods: ABT-yoghurts (Lactobacillus acidophilus La-5, Bifidobacterium animalis ssp. lactis BB-12, Streptococcus thermophilus) were produced from cow&rsquo;s milk with 0%, 5%, 10% or 15% (v/v) of Pu-erh tea infusion added before the fermentation stage. The products obtained were subjected to the following analyses one day after production (colour profile) and after 7, 14 and 21 days of cold storage: ferric reducing antioxidant power (FRAP) and anti-radical power (ARP) measured against DPPH radical, titratable acidity, pH, texture parameters (back extrusion test), viability of starter cultures and sensory quality (hedonic scale experiment)., Results: Pu-erh tea supplementation significantly enhanced the antioxidant potential of probiotic yoghurts as a 3&ndash;6.5-fold increase in FRAP and a 10&ndash;24-fold increase in ARP values were observed in comparison to plain ABT-yoghurt. Pu-erh tea slightly enhanced the viability of L. acidophilus and reduced the pH of probiotic yoghurts. Higher concentrations of Pu-erh tea caused decreased firmness and consistency while cohesiveness and index of viscosity remained unaffected upon supplementation. The addition of Pu-erh tea infusion modified the colour and sensory properties of the probiotic yoghurts but the sensory quality of the tea yoghurts was rated lower when compared to the plain one. Among all tea yoghurts, the one with 15% Pu-erh tea additive received the highest scores in sensory assessment., Conclusions: Pu-erh tea may be successfully applied as a functional additive to probiotic yoghurts, signifi- cantly enhancing the antioxidant properties of fermented milk and ensuring a high rate of starter bacteria viability during storage. However, the level of fortification must be carefully chosen as some doses negatively influence texture parameters and sensory quality.

Electrospun nanofiber membranes are widely investigated in the past few decades as candidates for tissue engineering, which can mimic natural extracellular matrix (ECM) and improve cell adhesion, proliferation, and expression on nanofiber membranes. However, the formation of bacterial biofilms on nanofiber membranes and application of the biofilm-integrated nanofiber membranes remain largely unknown. Here, electrospun cellulose acetate nanofiber membranes are first utilized as scaffold materials for Lactobacillus plantarum ( L. plantarum) biofilm formation. Nanofiber membranes proved to be an excellent scaffold for bacteria biofilm with high stability, where biofilms were interlocked with nanofibers forming a cohesive structure. In comparison with planktonic bacteria, L. plantarum biofilms on nanofiber membranes show excellent gastrointestinal resistance. Instead of decreasing, the number of viable cells increased after 3 h digestion in vitro. The L. plantarum biofilm-integrated nanofiber membranes were used as reusable starter cultures for fermented milk production showing excellent fermentative ability and higher survival of L. plantarum during shelf life. The viable cells in fermented milk remained at 11 log CFU/g throughout the reusable batches, which is far above the required value of 7 log CFU/g in commercial products. In addition, the produced fermented milk possesses shorter fermentation time and higher survival of probiotics during shelf life. The results suggest electrospun nanofiber membranes are ideal scaffold materials for bacteria biofilms immobilization in biotechnology and fermentation engineering, which broaden the potential use of electrospun nanofiber membranes in microbiology and strengthen the application of biofilms in fermentation engineering.

The current study investigates whether probiotic fermented milk (PFM) and yogurt consumption (YC) are related to both the ingested bacteria taxa and the overall gut microbiota (GM) composition in healthy adults. PFM and YC habits were analyzed in 260 subjects (51% male) by specific questionnaires, and the following groups were considered: (1) PFM groups: nonconsumers (PFM-NC, n = 175) and consumers (PFM, n = 85), divided as follows: Bifidobacterium -containing PFM (Bif-PFM; n = 33), Lactobacillus -containing PFM (Lb-PFM; n = 14), and mixed Bifidobacterium and Lactobacillus -containing PFM (Mixed-PFM; n = 38); (2) PFM-NC were classified as: yogurt nonconsumers (Y-NC; n = 40) and yogurt consumers (n = 135). GM was analyzed through 16S rRNA sequencing. PFM consumers showed higher Bifidobacteria taxa levels compared to NC, from phylum through to species. Specifically, Bif-PFM consumption was related to higher B. animalis levels ( p < 0.001), whereas Lb-PFM consumption was associated to higher levels of Bifidobacterium ( p < 0.045) and B. longum ( p = 0.011). YC was related to higher levels of the yogurt starter Streptococcus thermophilus ( p < 0.001). Lactobacilli and the overall GM were not related either to YC or PFM consumption. According to these results, healthy adults might benefit from PFM intake by increasing Bifidobacterium levels.

Lactic acid bacteria alone or with special adjunct probiotic strains are inevitable for the preparation of various specific functional foods. Moreover, because of their growth and metabolism, the final products are preserved for a certain time. Thus, growth dynamics of the lactic acid bacteria of the Fresco DVS 1010 culture ( Lactococcus lactis spp. lactis, Lactococcus lactis spp. cremoris, Streptococcus salivarius spp. thermophilus) during liquid-state fermentation of soya mashes and pH values within the process were analyzed in this study. Although milk is the most typical growth medium for the lactic acid bacteria, presumable viable counts of Fresco culture reached levels 10 9  CFU ml -1 after 8 h, representing 2-3 log increase in comparison to initial state (specific growth rates ranged from 1.06 to 1.64 h -1 ). After 21 days of storage period, the pH levels in the products were reduced to 4.50-4.70, representing a decrease of about 1.5-1.7 units. All prepared soybean products contained detectable amounts of raffinose-series oligosaccharides (0.25-0.68 g per 100 g) that were reduced in average by about 30.5% during period of 21 days.

The potential for reusability of whey is of concern due to its substantial nutritional value and, second, in view of the need to reduce environmental impact. The use of goat whey powder in the production of a fermented dairy beverage using a Lactobacillus casei culture as adjunct shows a good prospect of a food product with hypotensive activity. This study investigated the microbial viability, proteolysis and angiotensin-converting enzyme (ACE) inhibitory activity of a fermented dairy beverage produced with goat whey powder and a probiotic culture of L. casei BGP93 co-cultured with Streptococcus thermophilus TA-40. The probiotic beverage exhibited no significant difference from the control (absence of L. casei) with regard to titratable acidity, S. thermophilus viability and proteolysis degree during 21 days of storage (P > 0.05). During this period, the beverage maintained L. casei at appropriate levels (> 7 log cfu ml -1 ), thereby qualifying as a potential probiotic product. Although both control and probiotic beverages exhibited ACE inhibitory activity, as a result of proteolysis of whey proteins during fermentation, significant increased ACE inhibitory activity was found for the beverage with added probiotic L. casei (P < 0.05). The probiotic beverage has potency as a functional food candidate to be included in a dietary strategy aiming at prevention and control of hypertension.

Cold stress adversely affects cell viability and acidification, and new cryoprotective methods continue to be needed in cold-chain food industry. Given this, we investigated the cryoprotective effects and action mechanism of antifreeze peptides obtained from tilapia scales (TSAPP) on Streptococcus thermophilus during cold stress. Our results showed that the molecular weight of TSAPP ranged from 180 to 2000 Da and its thermal hysteresis activity was 0.29 °C. Growth of S. thermophilus was improved after treatment with TSAPP (1 mg/mL) under cold stress. This growth was notable when compared with the effects of other cryoprotectants. Furthermore, TSAPP improved the metabolic activity of S. thermophilus during cold stress. TSAPP likely offered its cellular protection by maintaining cell membrane fluidity through hydrogen bonding of the phospholipid bilayer. These results indicate that TSAPP has potential as a novel biological peptide material with cryoprotective activity for future use in probiotic or other processed food applications.

This study aimed to evaluate the profiles of Streptococcus thermophilus nutrient requirements to guide the design of media for high cell density culturing. The growth kinetics, physiological state, and nutrient requirement profiles of S. thermophilus were analyzed in chemically defined media. The results showed that the intracellular ATP concentration, H + -ATPase activity, NADH/NAD + , and NH 3 concentrations varied with intracellular pH. The nutrient components with the highest amounts required were Leu and Asp; ascorbic acid and p-amino benzoic acid; K + and PO 4 3- ; and guanine and uracil. The nutrient components with the largest required ratios were Arg, His, and Met; folic acid, cyanocobalamine, biotin, and nicotinic acid; Ca 2+ and Mg 2+ ; and guanine and uracil. In this study, different nutrient components were primarily used at different phase. Trp, Tyr, calcium pantothenate, thiamine, guanine, and Mg 2+ were mainly used from late-lag to midexponential phase. Met, Pro, Phe, Ala, Gly, nicotinic acid, and riboflavin were mainly used from midexponential to late-exponential phase. The highest bioavailabilities of nutrient components were also found at diverse phase. Met, Leu, Ile, Asn, Glu, Lys, Pro, Gly, riboflavin, nicotinic acid, adenine, uracil, inosine, and Ca 2+ had the highest bioavailability from late-lag to midexponential phase. Lactose, Glu, Asp, His, Trp, Cys, Val, Arg, Phe, Ala, Ser, Thr, Tyr, folate and cobalamin, calcium pantothenate, ascorbic acid, thiamine, biotin, p-amino benzoic acid, vitamin B 6 , K + , Mg 2+ , guanine, xanthine, and PO 4 3- had the highest bioavailability from midexponential to late-exponential phase. This study elucidated the nutrient requirement profiles with culture time and biomass at various average growth rates during the growth of S. thermophilus. The present results will help to formulate complex media for high cell density cultivation and provide the theoretical basis for S. thermophilus feeding strategies., (© 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

Several microbial modulatory concepts, such as certain probiotics and prebiotics, confer protection against gastrointestinal infections, among which is acute diarrhea caused by the rotavirus (RV). Other microbiota modulators, such as postbiotics, produced during fermentation, might also have the potential to counteract RV infection. In light of this, a fermented milk, made by using Bifidobacterium breve C50 (BbC50) and Streptococcus thermophilus 065 (St065) with a prebiotic mixture-short chain galactooligosaccharides/long chain fructooligosaccharides (scGOS/lcFOS 9:1)-with potential to impact the intestinal microbiota composition was tested. An RV infected rat model was used to evaluate the amelioration of the infectious process and the improvement of the immune response induced by the fermented milk with prebiotic mixture. The dietary intervention caused a reduction in the clinical symptoms of diarrhea, such as severity and incidence. Furthermore, a modulation of the immune response was observed, which might enhance the reduction of the associated diarrhea. In addition, the fermented milk with prebiotic mixture was able to bind the virus and reduce its clearance. In conclusion, the postbiotic components in the fermented milk in combination with the prebiotics used here showed protective properties against RV infection.

This study addresses the hypothesis that the extracellular cell-associated X-prolyl dipeptidyl-peptidase activity initially described in Streptococcus thermophilus could be attributable to the intracellular X-prolyl dipeptidyl-peptidase PepX. For this purpose, a PepX-negative mutant of S. thermophilus LMD-9 was constructed by interrupting the pepX gene and named LMD-9-ΔpepX. When cultivated, the S. thermophilus LMD-9 wild type strain grew more rapidly than its ΔpepX mutant counterpart. Thus, the growth rate of the LMD-9-ΔpepX strain was reduced by a factor of 1.5 and 1.6 in milk and LM17 medium (M17 medium supplemented with 2% lactose), respectively. The negative effect of the PepX inactivation on the hydrolysis of β-casomorphin-7 was also observed. Indeed, when incubated with this peptide, the LMD-9-ΔpepX mutant cells were unable to hydrolyze it, whereas this peptide was completely degraded by the S. thermophilus LMD-9 wild type cells. This hydrolysis was not due to leakage of intracellular PepX, as no peptide hydrolysis was highlighted in cell-free filtrate of wild type strain. Therefore, based on these results, it can be presumed that though lacking an export signal, the intracellular PepX might have accessed the β-casomorphin-7 externally, perhaps via its galactose-binding domain-like fold, this domain being known to help enzymes bind to several proteins and substrates. Therefore, the identification of novel distinctive features of the proteolytic system of S. thermophilus will further enhance its credibility as a starter in milk fermentation., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background and Objective: Hepatorenal syndrome (HRS) is a major public health problem in which both liver and kidney dysfunctions are encountered. The present research aimed to investigate the beneficial use of micro-encapsulated probiotic alone (Bifidobacterium bifidum, Lactobacillus delbrueckii and Streptococcus thermophilus mixture) or with green tea alcohol extract in HRS model in rats., Materials and Methods: Flavonoids content and in vitro antioxidant activity of the extract were assessed. The animal experiment consisted of 4 groups; control healthy, control with HRS and two test groups with HRS and treated with either the encapsulated probiotic mixture alone or with green tea extract. After 3 weeks; urinary creatinine was determined in 24 h rat urine samples. Colonic microbiota was assessed in faeces. Plasma malondialdehyde, nitrite, C-reactive protein, creatinine, uric acid, urea and the activity of transaminases, catalase (CAT) and angiotensin-1 converting enzyme (ACE-1) were determined with calculation of creatinine clearance., Results: Results showed significant increase in all biochemical parameters of HRS control except for ACE-1, CAT and creatinine clearance that experienced significant reduction along with dysbiosis compared to healthy control. Test groups showed improvement in all biochemical parameters with superiority to probiotic-green tea extract combination. Both treatments produced significant increase in fecal B. bifidum, S. thermophilus and L. bulgaricus and reduction of Staphylococci and Coliform. The effect of probiotic-green tea extract combination was more pronounced concerning the last three. Flavonoids and antioxidant activity of the extract were 1.325±0.01 mg quercetin/g and 98±1.66%, respectively., Conclusion: Administration of micro-encapsulated probiotic with or without alcohol green tea extract exerted significant prevention of HRS in rat with superiority to probiotic-green tea extract combination.

Within the genus Streptococcus, S. thermophilus and S. macedonicus are the 2 known species related to foods. Streptococci are widely used as starter cultures to rapidly lower milk pH. As S. macedonicus has been introduced quite recently, much less information is available on its technological potential. Because temperature is an important factor in fermented food production, we compared the growth kinetics over 24 h of 8 S. thermophilus and 7 S. macedonicus strains isolated from various dairy environments in Italy, at 4 temperatures, 30°C, 34°C, 37°C and 42°C. We used the Gompertz model to estimate the 3 main growth parameters; namely, lag phase duration (λ), maximum growth rate (µ max ), and maximum cell number at the stationary phase (N max ). Our results showed significant differences in average growth kinetics between the 2 species. Among the strains tested, 37°C appeared to be the optimal temperature for the growth of both species, particularly for S. macedonicus strains, which showed mean shorter lag phases and higher cell numbers compared with S. thermophilus. Overall, the growth curves of S. macedonicus strains were more similar to each other whereas S. thermophilus strains grew very differently. These results help to better define and compare technological characteristics of the 2 species, in view of the potential use of S. macedonicus in place of S. thermophilus in selected technological applications., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The lactic acid bacterium Streptococcus thermophilus is a major starter culture for the production of dairy products. In this study, the physiochemical characteristics of milk fermented by the MGA45-4 isolate of S. thermophilus were analyzed. Our data indicate that milk fermented using S. thermophilus MGA45-4 maintained a high viable cell count (8.86 log10 colony-forming units/mL), and a relatively high pH (4.4), viscosity (834.33 mPa·s), and water holding capacity (40.85%) during 14 days of storage. By analyzing the volatile compound profile using solid-phase microextraction and gas chromatography/mass spectrometry, we identified 73 volatile compounds in the fermented milk product, including five carboxylic acids, 21 aldehydes, 13 ketones, 16 alcohols, five esters, and 13 aromatic carbohydrates. According to the odor activity values, 11 of these volatile compounds were found to play a key role in producing the characteristic flavor of fermented milk, particularly octanal, nonanal, hexanal, 2,3-butanedione, and 1-octen-3-ol, which had the highest odor activity values among all compounds analyzed. These findings thus provide more insights in the chemical/molecular characteristics of milk fermented using S. thermophilus , which may provide a basis for improving dairy product flavor/odor during the process of fermentation and storage., Competing Interests: The authors declare no conflicts of interest.

Soymilk was produced from vegetable soybean and fermented by probiotics (Lactobacillus acidophilus La-5, Bifidobacterium animalis Bb-12) in co-culture with Streptococcus thermophilus. The composition of the fermented beverage and oligosaccharides content were determined. The effect of fructooligosaccharides and inulin on the fermentation time and viability of probiotic microorganisms throughout 28 days of storage at 5°C were evaluated. The soymilk from vegetable soybeans was fermented in just 3.2h, when pH reached 4.8. Fermentation reduced the contents of stachyose and raffinose in soymilk. Prebiotics had no effect on acidification rate and on viability of B. animalis and S. thermophilus in the fermented beverage. The viable counts of B. animalis Bb-12 remained above 10 8 CFUmL -1 in the fermented soymilk during 28 days of storage at 5°C while L. acidophilus La-5 was decreased by 1logCFUmL -1 . The fermented soymilk from vegetable soybeans showed to be a good food matrix to deliver probiotic bacteria, as well as a soy product with a lower content of non-digestible oligosaccharides., (Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

The ability of folate-producer strains of Streptococcus thermophilus to accumulate folates and the expression of two target genes (folK and folP), involved in the folate biosynthesis, were studied during milk fermentation. An over-expression of folK took place only in the early phase of growth, whereas folP was mainly expressed in the mid log-phase of growth and declined thereafter. The accumulation of total folates, which was quantified by a microbiological assay, was strain-dependent. Two major forms of folates, i.e. tetrahydrofolate (THF) and 5-methyl-tetrahydrofolate (5-Met-THF), were identified and quantified by HPLC. With respect to the level accumulated by a weak folate producer (St 383), used as calibrator in the expression experiments and as control in folate quantification in milk, the strains St 563 and St 399 produced 5-Met-THF in amounts significantly higher than THF. The possibility of using selected folate-producer S. thermophilus strains as functional cultures for a bio-fortification of dairy products is discussed., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

This study investigated the differential effect of salt concentration in the outside and inside layers of brine salted cheeses on viability, culturability and enzyme activity of starter bacteria. The high-salt environment of the outside layer caused a sharp decrease in L. helveticus viability as measured by traditional plate counts. Remarkably, this was associated with lower release of intracellular enzymes (LDH), reduced levels of proteolysis and larger membrane integrity as measured by flow cytometry (FC) following classical Live/Dead staining. FC analysis of light scattering properties highlighted a significant reduction in size and granularity of the microbiota located in the cheese surface, suggestive of cell shrinkage and condensation of internal macromolecules probably due to hyperosmotic stress. The microbiota of the cheese surface were found to experience greater oxidative stress, as measured by FC analysis of the total levels of reactive oxygen species, compared to that of the interior layer. These results lead us to postulate that the physiology and health status of the microbiota were significantly different in the outer and inner layers of the cheese. The hyperosmotic environment of the outer layer resulted in reduced cell lysis, as measurable by assays based upon membrane integrity, but rather triggered cell death via mechanisms involving cell shrinkage and ROS-mediated damage of vital intracellular components. This study challenges the current thinking on how salt controls microbial activity in ripening cheese, especially in cheeses which are brine salted as local variations in biochemical ripening indices can differ significantly from the outside to the inside of a ripening cheese., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The contribution of two starter (Lactobacillus delbrueckii and Streptococcus thermophilus) and nine non-starter (Enterococcus casselliflavus, Enterococcus faecalis, Enterococcus durans, Enterococcus gallinarum, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, Pediococcus acidilactici and Pediococcus pentosaceus) species of lactic acid bacteria (LAB) to the volatile organic compounds (VOCs) of Caciocavallo Palermitano cheese was investigated. The strains used in this study were isolated during the production/ripening of the stretched cheese and tested in a cheese-based medium (CBM). The fermented substrates were analyzed for the growth of the single strains and subjected to the head space solid phase micro-extraction (HS-SPME) and gas chromatography - mass spectrometry (GC-MS). The 11 strains tested were all able to increase their numbers in CBM, even though the development of the starter LAB was quite limited. GC-MS analysis registered 43 compounds including seven chemical classes. A lower diversity of VOCs was registered for the unfermented curd based medium (CuBM) analyzed for comparison. The class of ketones represented a consistent percentage of the VOCs for almost all LAB, followed by alcohols and esters. The volatile profile of Pediococcus acidilactici and Lactobacillus delbrueckii was mainly characterized by 2-butanol, butanoic acid and hexanoic acid and their esters, while that of Lactobacillus casei and Lactobacillus rhamnosus was characterized by 2,3-butanedione and 2-butanone, 3-hydroxy. In order to correlate the VOCs produced by Caciocavallo Palermitano cheeses with those generated by individual LAB, the 4-month ripened cheeses resulting from the dairy process monitored during the isolation of LAB were also analyzed for the volatile chemical fraction and the compounds in common were subjected to a multivariate statistical analysis. The canonical analysis indicated that the VOCs of the ripened cheeses were mainly influenced by E. gallinarum, L. paracasei, L. delbrueckii, L. rhamnosus and L. casei and that 1-hexanol, o-xylene and m-xylene were the cheese VOCs highly correlated with LAB., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The effect of W 1 /O/W 2 emulsion incorporation in set-type yogurt on the acidification process, physicochemical properties, bacterial growth kinetics and structural characteristics was investigated. The W 1 /O/W 2 emulsion was formed by using a two-step homogenisation process and milk as the W 1 and W 2 phases, and stability was monitored with optical microscopy and cryo-SEM. Adding the W 1 /O/W 2 emulsions reduced the acidification rate, viscosity and water retention capacity. Texture (adhesiveness, cohesiveness, hardness, and gumminess) differed in yogurts containing W 1 /O/W 2 emulsion compared to controls during the acidification process, however, trends became stable during storage. The growth of S. thermophilus during the acidification process of yogurt was reduced in the presence of W 1 /O/W 2 emulsion while L. bulgaricus trended higher during storage. This study shows that yogurts containing W 1 /O/W 2 emulsion are feasible subject to processing modification., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Dairy desserts have emerged as interesting options for the incorporation of probiotics, bioactive ingredients and alternative sources of thickeners. This shows an opportunity to investigate the use of Gracilaria seaweeds in the formulation of potentially probiotic dairy desserts. This study aimed to compare the effects of dispersions obtained from Gracilaria domingensis and Gracilaria birdiae used as thickening agents on texture properties of dairy desserts fermented with SAB 440-A, composed of the starter Streptococcus thermophilus and the potential probiotics Bifidobacterium animalis and Lactobacillus acidophilus, and also to study their physicochemical characteristics, microbial viability and sensory acceptability. No significant differences between desserts with G. birdiae or G. domingensis dispersions regarding total solids, ash and fat content, as well as pH, titratable acidity, the viability of the microorganisms of the mixed culture and sensory acceptability were verified (P > 0.05). Nonetheless, the dessert with G. domingensis dispersion showed higher dietary fibre content and significantly increased firmness than the one produced with G. birdiae (P < 0.05). Moreover, B. animalis was able to maintain higher populations, above 7 log cfu g -1 during 21 days of storage of desserts, in the presence of either G. birdiae or G. domingensis dispersions, despite the fact that L. acidophilus has shown low viability in the final products. Therefore, the G. domingensis dispersion is suitable to be used as a thickening agent to produce dairy desserts with enhanced firmness and good sensory acceptability, it being also advisable to use only B. animalis as a probiotic for this product.

Background: The promotion effect on proliferation of Streptococcus thermophilus by enzymatic hydrolysates of aquatic products was firstly studied. The effect of influencing factors of the hydrolysis on the growth of S. thermophilus was investigated., Result: Grass Carp fish skin was hydrolysed to peptides by enzymatic hydrolysis using protease ProteAX, and for the S. thermophilus growth, the optimal enzymatic hydrolysis conditions were temperature of 60 °C, initial pH of 9.0, enzyme concentration of 10 g kg -1 , hydrolysis time of 80 min, and ratio of material to liquid of 1:2. The Grass Carp fish skin hydrolysate (GCFSH) prepared under the optimum conditions was fractionated to five fragments (GCFSH 1, GCFSH 2, GCFSH 3, GCFSH 4, GCFSH 5) according to molecular weight sizes, in which the fragments GCFSH 4 and GCFSH 5, with molecular weights of less than 1000 Da, significantly promoted the growth of S. thermophilus., Conclusion: The hydrolysis process of Grass Carp fish skin can be simplified, and the peptides with molecular weights below 1000 Da in the hydrolysates are the best nitrogen source for proliferation of S. thermophilus. This work can provide a fundamental theoretical basis for the production of multi-component functional foods, especially in milk drinks or yogurt. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

Microencapsulation is an effective technology used to protect probiotics against harsh conditions. Extrusion is a commonly used microencapsulation method utilized to prepare probiotics microcapsules that is regarded as economical and simple to operate. This research aims to prepare acid-resistant probiotic microcapsules with high viability after freeze-drying and optimized storage stability. Streptococcus thermophilus IFFI 6038 (IFFI 6038) cells were mixed with trehalose and alginate to fabricate microcapsules using extrusion. These capsules were subsequently coated with chitosan to obtain chitosan-trehalose-alginate microcapsules with shell-matrix structure. Chitosan-alginate microcapsules (without trehalose) were also prepared using the same method. The characteristics of the microcapsules were observed by measuring the freeze-dried viability, acid resistance, and long-term storage stability of the cells. The viable count of IFFI 6038 in the chitosan-trehalose-alginate microcapsules was 8.34 ± 0.30 log CFU g -1 after freeze-drying (lyophilization), which was nearly 1 log units g -1 greater than the chitosan-alginate microcapsules. The viability of IFFI 6038 in the chitosan-trehalose-alginate microcapsules was 6.45 ± 0.09 log CFU g -1 after 120 min of treatment in simulated gastric juices, while the chitosan-alginate microcapsules only measured 4.82 ± 0.22 log CFU g -1 . The results of the long-term storage stability assay indicated that the viability of IFFI 6038 in chitosan-trehalose-alginate microcapsules was higher than in chitosan-alginate microcapsules after storage at 25 °C. Trehalose played an important role in the stability of IFFI 6038 during storage. The novel shell-matrix chitosan-trehalose-alginate microcapsules showed optimal stability and acid resistance, demonstrating their potential as a delivery vehicle to transport probiotics., (© 2017 Institute of Food Technologists®.)

The proto-cooperation between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus in the yogurt consortium enhances the growth rate and size of each population. In contrast, the independent growth of the two species in milk leads to a slower growth rate and a smaller population size. In this study, we report the first evidence that the urease activity of S. thermophilus increases the intracellular pH of L. delbrueckii in the absence of carbon source. However, in milk, in the presence of lactose the alkalizing effect of urea-derived ammonia was not detectable. Nevertheless, based on glucose consumption and lactic acid production at different pH in , L. delbrueckii showed an optimum of glycolysis and homolactic fermentation at alkaline pH values. In milk, we observed that ammonia provided by urea hydrolysis boosted lactic acid production in S. thermophilus and in L. delbrueckii when the species were grown alone or in combination. Therefore, we propose that urease activity acts as an altruistic cooperative trait, which is costly for urease-positive individuals but provides a local benefit because other individuals can take advantage of urease-dependent ammonia release., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In Streptococcus thermophilus , gene transfer events and loss of ancestral traits over the years contribute to its high level of adaptation to milk environments. Biofilm formation capacity, a phenotype that is lost in the majority of strains, plays a role in persistence in dairy environments, such as milk pasteurization and cheese manufacturing plants. To investigate this property, we have studied S. thermophilus UC8547, a fast-acidifying dairy starter culture selected for its high capacity to form biofilm on stainless steel under environmental conditions resembling the dairy environment. Using a dynamic flow cell apparatus, it was shown that S. thermophilus UC8547 biofilm formation on stainless steel depends on the presence of milk proteins. From this strain, which harbors the prtS gene for the cell wall protease and shows an aggregative phenotype, spontaneous mutants with impaired biofilm capacity can be isolated at high frequency. These mutants lack the PrtS expendable island, as confirmed by comparison of the genome sequence of UC8547Δ3 with that of the parent strain. The prtS island excision occurs between two 26-bp direct repeats located in the two copies of the IS Sth1 flanking this genomic island. The central role of PrtS was confirmed by analyzing the derivative strain UC8547Δ16, whose prtS gene was interrupted by an insertional mutation, thereby making it incapable of biofilm formation. PrtS, acting as a binding substance between the milk proteins adhered to stainless steel and S. thermophilus cell envelopes, mediates biofilm formation in dairy environments. This feature provides S. thermophilus with an ecological benefit for its survival and persistence in this environment. IMPORTANCE The increased persistence of S. thermophilus biofilm has consequences in the dairy environment: if, on the one hand, the release of this microorganism from biofilm can promote the fermentation of artisanal cheeses, under industrial conditions it may lead to undesirable contamination of dairy products. The study of the molecular mechanism driving S. thermophilus biofilm formation provides increased knowledge on how an ancestral trait affects relevant phenotypes, such as persistence in the environment and efficiency of growth in milk. This study provides insight into the genetic factors affecting biofilm formation at dairy plants., (Copyright © 2017 American Society for Microbiology.)

A set of 191 strains of Streptococcus thermophilus were preliminarily screened for the presence of the genes codifying for cell envelope-associated proteinase (prtS) and for glutamate decarboxylase (gadB) responsible for γ-aminobutyric acid (GABA) production. The growth and proteolytic activity of the gadB-positive strains (9 presenting the prtS gene and 11 lacking it) were studied in microfiltered pasteurized milk. Degradation of both caseins (capillary electrophoresis) and soluble nitrogen fractions (HPLC) and changes in the profile of free amino acids (FAAs; ion-exchange chromatography) were evaluated at inoculation and after 6 and 24 h of incubation at 41 °C. None of the strains was capable of hydrolyzing caseins and β-lactoglobulin, and only two hydrolyzed part of α-lactalbumin, these proteins being present in their native states in pasteurized milk. Contrarily, most strains were able to hydrolyze peptones and peptides. For initial growth, most strains relied on the FAAs present in milk, whereas, after 6 h, prtS + strains released variable amounts of FAA. One prtS + strain expressed a PrtS - phenotype, and two prtS - strains showed a rather intense proteolytic activity. Only five strains (all prtS + ) produced GABA, in variable quantities (up to 100 mg/L) and at different rates, depending on the acidification strength. Addition of glutamate did not induce production of GABA in nonproducing strains that, however, unexpectedly were shown to adopt the degradation of arginine into citrulline and ornithine as an alternative acid resistance system and likely as a source of ATP.

Objective: To test if the production of bacteriocins by Streptococcus thermophilus is influenced when grown in various complex media commonly used for the culturing of lactic acid bacteria., Results: Forty-one strains of S. thermophilus were screened for the production of bacteriocins in tryptone/yeast extract/lactose (TYL), M17-lactose (M17L), M17-glucose (M17G) and MRS media. Two strains, ST144 and ST145, were identified as novel bacteriocin producers, with constitutive production observed only in M17G. Strains ST110, ST114 and ST134 constitutively produced bacteriocins in all growth media but ST114 required growth in MRS for its antimicrobial activity to persist in a 24 h culture. The addition of a synthetic quorum sensing peptide (BlpC) induced bacteriocin production by ST106 in all media tested; and by ST118 in TYL and M17L. Strain ST109, which constitutively produced a bacteriocin in TYL and M17 broths, required BlpC induction when grown in MRS. Real-time PCR analysis showed that the natural expression of blpC in ST109 was lower when grown in MRS, suggesting that something in medium interfered with the blp quorum sensing system., Conclusion: As the choice of growth medium influences both bacteriocin production and peptide stability, several types of production media should be tested when screening for novel bacteriocin-producing strains of S. thermophilus.

Background: Probiotic supplementation to women during pregnancy and lactation can modulate breast milk composition, with immune benefits being transferred to their infants., Aim: The aim of the study was to evaluate the effect of high-dose probiotic supplementation to women during late pregnancy and lactation on cytokine profile and secretory IgA (sIgA) in breast milk and thus to study if differences in breast milk composition can affect lactoferrin and sIgA levels in stool samples of newborns. The safety of maternal probiotic administration on neonatal growth pattern and gastrointestinal symptoms were also evaluated., Methods: In a double-blind, placebo-controlled, randomized trial, 66 women took either the probiotic ( n = 33) or a placebo ( n = 33) daily. Levels of interleukins (IL-6, IL-10 and IL-1β), transforming growth factor-β1 (TGF-β1), and sIgA in breast milk; and the level of sIgA and lactoferrin in newborn stool samples were analyzed at birth and then again at one month of life. Antropometrical evaluation and analysis of gastrointestinal events in newborns was also performed., Results: Probiotic maternal consumption had a significant impact on IL6 mean values in colostrum and on IL10 and TGF-β1 mean values in mature breast milk. Fecal sIgA mean values were higher in newborns whose mothers took the probiotic product than in the control group. Probiotic maternal supplementation seems to decrease incidence of infantile colic and regurgitation in infants., Conclusion: High-dose multi-strain probiotic administration to women during pregnancy influences breast milk cytokines pattern and sIgA production in newborns, and seems to improve gastrointestinal functional symptoms in infants., Competing Interests: The authors declare no conflicts of interest.

Aims: The aim of this study was to elucidate the effect of arginine on the growth, arginine metabolism and amino acid consumption profiles of Streptococcus thermophilus T1C2., Methods and Results: The growth kinetics, intracellular pH, extracellular osmotic pressure, expression of key genes in the arginine metabolism pathway and amino acid consumption profiles were analysed in chemically defined medium with different initial arginine concentrations. The results showed that arginine stimulated the growth of Strep. thermophilus T1C2 under low intracellular pH and high extracellular osmotic pressure. The expression of key genes in the arginine degradation pathway indicated that arginine relieved the drop in the intracellular pH by consuming protons and generating NH3 . Additionally, the results showed that arginine degradation did not occur via the arginine deiminase pathway but through the arginine decarboxylase-urease pathway. Furthermore, the utilization efficiency of amino acids was improved in the presence of arginine., Conclusions: Arginine improved the growth of Strep. thermophilus due to protecting Strep. thermophilus against intracellular acid stress, which was revealed at the transcriptional level of key genes. This study showed that the acid resistance of Strep. thermophilus was achieved through the arginine decarboxylase-urease pathway., Significance and Impact of the Study: The arginine-stimulated growth of Strep. thermophilus improved the utilization efficiency of amino acids and reduced nitrogen waste, which could be useful for the optimization of cultivation media., (© 2016 The Society for Applied Microbiology.)

Background: White mold-ripened cheeses were investigated with the objective of proposing a colorimetric method to monitor the surface growth of Penicillium candidum and to evaluate the influence of the mesophilic (homofermentative (QMO) and heterofermentative (QMLD)) and thermophilic (QT) starter cultures on the physicochemical composition and sensory description., Results: The whiteness index was effective in proving the appearance of superficial mycelium and the stability of white mold growth. The lactic cultures showed significant influence on most of the physicochemical analyses. The cheese made with thermophilic lactic culture had a 1 day gain in the growth of mycelium on the surface; nevertheless, the appearance of this product was potentially not acceptable for consumers. The heterofermentative mesophilic cheese had a better appearance and texture profile. However, the homofermentative mesophilic cheese showed aspects of fresh cheese and was acceptable for a wide range of consumers., Conclusion: The whiteness index was efficient to monitor the surface growth of P. candidum. The highest proteolytic effect was found in the QMLD and QT cultures. However, the cheese elaborated with the QMLD culture showed the best sensory acceptance. © 2015 Society of Chemical Industry., (© 2015 Society of Chemical Industry.)

The objective of this study was to investigate the effect of 2 plant leaf extracts on fermentation mechanisms and health-promoting activities and their potential as a nutraceutical prebiotics ingredient for application in dairy products. The individual active phenolic compounds in the plant extract-supplemented milk and yogurts were also identified. Compared with control fermentation, the plant extracts significantly increased the growth and acidification rate of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. In particular, plant extract components, including monosaccharides, formic acid, and hydroxycinnamic acid, such as neo-chlorogenic, chlorogenic, and caffeic acid, together play a stimulatory role and cause this beneficial effect on the growth of yogurt culture bacteria through fermentation. In addition, supplementation with the plant extracts enhanced antioxidant activities with increased total phenolic contents, especially the highest antioxidant activity was observed in yogurt supplemented with Cudrania tricuspidata leaf extract., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Electron microscopy was used to test whether or not (a) in statu nascendi synthesized, and in situ measured, nanoparticle size does not differ significantly from the size of nanoparticles after their purification; and (b) the generation of selenium is detrimental to the bacterial strains that produce them. Elemental nano-sized selenium produced by probiotic latic acid bacteria was used as a lactomicroselenium (lactomicroSel) inhibitor of cell growth in the presence of lactomicroSel, and was followed by time-lapse microscopy. The size of lactomicroSel produced by probiotic bacteria was measured in situ and after isolation and purification. For these measurements the TESLA BS 540 transmission electron microscope was converted from analog (aTEM) to digital processing (dTEM), and further to remote-access internet electron microscopy (iTEM). Lactobacillus acidophilus produced fewer, but larger, lactomicroSel nanoparticles (200-350 nm) than Lactobacillus casei (L. casei), which generated many, smaller lactomicroSel particles (85-200 nm) and grains as a cloudy, less electrodense material. Streptococcus thermophilus cells generated selenoparticles (60-280 nm) in a suicidic manner. The size determined in situ in lactic acid bacteria was significantly lower than those measured by scanning electron microscopy after the isolation of lactomicroSel particles obtained from lactobacilli (100-500 nm), but higher relative to those isolated from Streptococcus thermopilus (50-100 nm). These differences indicate that smaller lactomicroSel particles could be more toxic to the producing bacteria themselves and discrepancies in size could have implications with respect to the applications of selenium nanoparticles as prebiotics.

The effects of the Gracilaria domingensis seaweed aqueous extract in comparison with gelatin on the physicochemical, microbial, and textural characteristics of fermented milks processed with the mixed culture SAB 440 A, composed of Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium animalis ssp. lactis, were investigated. The addition of G. domingensis aqueous extract did not affect pH, titratable acidity, and microbial viability of fermented milks when compared with the control (with no texture modifier) and the products with added gelatin. Fermented milk with added the seaweed aqueous extract showed firmness, consistency, cohesiveness, and viscosity index at least 10% higher than those observed for the control product (P < 0.05). At 4 h of fermentation, the fermented milks with only G. domingensis extract showed a texture comparable to that observed for products containing only gelatin. At 5 h of fermentation, firmness and consistency increased significantly (P < 0.05) in products with only seaweed extract added, a behavior not observed in products with the full amount of gelatin, probably due to the differences between the interactions of these ingredients with casein during the development of the gel network throughout the acidification of milk. The G. domingensis aqueous extract appears as a promising gelatin alternative to be used as texture modifier in fermented milks and related dairy products., (© 2016 Institute of Food Technologists®)

This study evaluated the effects of sodium chloride reduction and its substitution with potassium chloride on Akawi cheese during storage for 30 d at 4 °C. Survival of probiotic bacteria (Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium longum) and starter bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus), angiotensin-converting enzyme-inhibitory and antioxidant activities, and concentrations of standard amino acids as affected by storage in different brine solutions (10% NaCl, 7.5% NaCl, 7.5% NaCl+KCl [1:1], 5% NaCl, and 5% NaCl+KCl [1:1]) were investigated. Furthermore, viability of human colon cells and human colon cancer cells as affected by the extract showing improved peptide profiles, highest release of amino acids and antioxidant activity (that is, from cheese brined in 7.5% NaCl+KCl) was evaluated. Significant increase was observed in survival of probiotic bacteria in cheeses with low salt after 30 d. Calcium content decreased slightly during storage in all cheeses brined in various solutions. Further, no significant changes were observed in ACE-inhibitory activity and antioxidant activity of cheeses during storage. Interestingly, concentrations of 4 essential amino acids (phenylalanine, tryptophan, valine, and leucine) increased significantly during storage in brine solutions containing 7.5% total salt. Low concentration of cheese extract (100 μg/mL) significantly improved the growth of normal human colon cells, and reduced the growth of human colon cancer cells. Overall, the study revealed that cheese extracts from reduced-NaCl brine improved the growth of human colon cells, and the release of essential amino acids, but did not affect the activities of potential bioactive peptides., (© 2016 Institute of Food Technologists®)