New Wild-Type Lacticaseibacillus rhamnosus Strains as Candidates to Manage Type 1 Diabetes.

The incidence of type 1 diabetes (T1D) has been dramatically increased in developed countries, and beyond the genetic impact, environmental factors, including diet, seem to play an important role in the onset and development of the disease. In this vein, five Lacticaseibacillus rhamnosus, isolated from traditional fermented Greek products, were screened for potential probiotic properties, aiming at maintaining gut homeostasis and antidiabetic capability to alleviate T1D symptoms. L. rhamnosus cell-free supernatants induced strong growth inhibitory activity against common food spoilage and foodborne pathogenic microorganisms, associated with several diseases, including T1D, and were also able to inhibit α-glucosidase activity (up to 44.87%), a promising property for alternatives to the antidiabetic drugs. In addition, survival rates up to 36.76% were recorded during the application of the static in vitro digestion model. The strains had no hemolytic activity and were sensitive to common antibiotics suggested by the European Food and Safety Association, apart from chloramphenicol. However, it is highly unlikely that the resistance has been acquired. In conclusion, our results suggest a great health-promoting potential of the newly isolated wild-type L. rhamnosus strains, but further confirmation of their efficiency in experimental animal models is considered an essential next research step.

Effect of cellulose crystallinity modification by starch gel treatment for improvement in ethanol fermentation rate by non-GM yeast cell factories.

This paper studies the reduction of crystallinity degree (CD) of cellulose treated with starch gel (SG), and the correlation of CD with the fermentation efficiency of cellulose to fuel-grade ethanol. Cellulose bioconversion from wood sawdust, consisting of three processes, was conducted in the same batch (one-step). The XRD and TEM analysis revealed 11% reduction in cellulose CD after its treatment with SG. One-step bioconversion process was performed employing two cell factories (CF) of non-engineered S. cerevisiae. CFs contained non- engineered S. cerevisiae cells covered with either SG entrapping Trichoderma reesei or cellulases prepared in the laboratory and immobilized in SG. The consolidated fermentation of treated cellulose resulted in an increase of bioethanol concentration (60-90%) in 2-day fermentation and the maximum ethanol concentration reached was approximately 5 mL/L (3.95 g/L). The fermentation efficiency for grade-fuel ethanol production was improved by cellulose pretreatment using SG to achieve reduced CD.

Health Promoting Properties of Cereal Vinegars.

Vinegar has been used for its health promoting properties since antiquity. Nowadays, these properties are investigated, scientifically documented, and highlighted. The health benefits of vinegar have been associated with the presence of a variety of bioactive components such as acetic acid and other organic acids, phenolic compounds, amino acids, carotenoids, phytosterols, vitamins, minerals, and alkaloids, etc. These components are known to induce responses in the human body, such as antioxidant, antidiabetic, antimicrobial, antitumor, antiobesity, antihypertensive, and anti-inflammatory effects. The diversity and levels of bioactive components in vinegars depend on the raw material and the production method used. Cereal vinegars, which are more common in the Asia-Pacific region, are usually made from rice, although other cereals, such as millet, sorghum, barley, malt, wheat, corn, rye, oats, bran and chaff, are also used. A variety of bioactive components, such as organic acids, polyphenols, amino acids, vitamins, minerals, alkaloids, melanoidins, butenolides, and specific compounds such as γ-oryzanol, tetramethylpyrazine, γ-aminobutyric acid, etc., have been associated with the health properties of cereal vinegars. In this work, the bioactive components and the related health effects of cereal vinegars are reviewed, and the most recent scientific literature is presented and discussed.

Yogurts Supplemented with Juices from Grapes and Berries.

Nowadays, there is growing interest for the development of enriched dairy products with phenolic compounds derived from edible sources, mainly due to their safety and potential health benefits. Following that trend, in the present study, fruit juices (blueberry, aronia, and grape) were supplemented into yogurt as functional ingredients. The main physicochemical characteristics (pH, reducing sugars, acidity, color, and syneresis), total phenolic content, antioxidant activity, and viability of yogurt starters were monitored during production and storage. The use of juices had no significant effect on milk acidification rate and on the main physicochemical characteristics of yogurts, while resulted in increased red color. Total phenolic content increased from 30 to 33% (grape and aronia) and up to 49% (blueberry), while similar results were observed in antioxidant activity. Similar values of syneresis were presented in all yogurts, probably due to exopolysaccharide producing starter culture. Streptococcus thermophilus retained high viable counts during storage especially in yogurts with fruit juices (>108 cells g-1) revealing a possible prebiotic effect of juices. The results obtained from this study show that fruit juices (aronia, blueberry, and grape) have potential to be used in yogurt production in order to optimize the benefits of probiotic products with high phenolic compound intake.

Assessment of Freeze-Dried Immobilized Lactobacillus casei as Probiotic Adjunct Culture in Yogurts.

Freeze-dried immobilized Lactobacillus casei ATCC 393 on casein and apple pieces were assessed as a probiotic adjunct culture for novel probiotic yogurt production. The effect of probiotic culture on physicochemical characteristics, probiotic cell survival, volatile aroma compounds, and sensory quality were evaluated during 28 days of storage at 4 °C. The use of L. casei resulted in lower pH values (3.92-4.12), higher acidity (0.88-1.10 g lactic acid/100 g of yogurt), and lower syneresis (40.8%-42.6%) compared to traditionally produced yogurt (pH 4.29; acidity 0.83 g lactic acid/100 g of yogurt; syneresis 44.1%). Microbiological and strain-specific multiplex PCR (Polymerase Chain Reaction) analysis confirmed that immobilized L. casei ATCC 393 cells were detected in yogurts at levels >7 log cfu g-1 after 28 days. In addition, probiotic supplementation significantly affected the concentrations of key volatile compounds, like acetic and other organic acids, 2-ethyl-1-hexanol, acetoin, and 2-butanone, as revealed by GC-MS (Gas Chromatography-Mass Spectrometry) analysis. Finally, the sensory evaluation demonstrated that the new products exhibited improved characteristics compared to traditionally produced yogurts.

Effect of Milk Type on the Microbiological, Physicochemical and Sensory Characteristics of Probiotic Fermented Milk.

The production of fermented milk using cow milk, goat milk and a mixture of them (1:1) was evaluated. The traditional yogurt starter culture was used-Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus, along with Lactobacillus casei ATCC 393. The objective was to monitor the viability of these cultures during the storage of fermented milk at 4 °C for up to 28 days. Storage significantly affected the viability of all microorganisms and especially L. bulgaricus. All microorganisms retained viable numbers higher than 7.5 log CFU g-1, even after 4 weeks of storage, which is important to confirm the probiotic character of the product. The use of goat milk significantly affected the pH and acidity of fermented milk. More specifically, the use of goat milk led to fermented milk with lower pH values and higher acidities than fermented milk with cow milk. The sensory evaluation ascertained the overall quality of fermented milk with a mixture of cow and goat milk, which scored similar values to the commercial sample.

Technological Development of Brewing in Domestic Refrigerator Using Freeze-Dried Raw Materials.

Development of a novel directly marketable beer brewed at low temperature in a domestic refrigerator combined with yeast immobilization technology is presented in this study. Separately, freeze-dried wort and immobilized cells of the cryotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 on tubular cellulose were used in low-temperature fermentation (2, 5 and 7 °C). The positive effect of tubular cellulose during low-temperature brewing was examined, revealing that freeze-dried immobilized yeast cells on tubular cellulose significantly reduced the fermentation rates in contrast to freeze-dried free cells, although they are recommended for home-made beer production. Immobilization also enhanced the yeast resistance at low-temperature fermentation, reducing the minimum brewing temperature value from 5 to 2 °C. In the case of high-quality beer production, the effect of temperature and initial sugar concentration on the fermentation kinetics were assessed. Sensory enrichment of the produced beer was confirmed by the analysis of the final products, revealing a low diacetyl concentration, together with improved polyphenol content, aroma profile and clarity. The proposed process for beer production in a domestic refrigerator can easily be commercialized and applied by dissolving the content of two separate packages in tap water; one package containing dried wort and the other dried immobilized cells on tubular cellulose suspended in tap water.

Lactobacillus casei Exerts Anti-Proliferative Effects Accompanied by Apoptotic Cell Death and Up-Regulation of TRAIL in Colon Carcinoma Cells.

Probiotic microorganisms such as lactic acid bacteria (LAB) exert a number of strain-specific health-promoting activities attributed to their immunomodulatory, anti-inflammatory and anti-carcinogenic properties. Despite recent attention, our understanding of the biological processes involved in the beneficial effects of LAB strains is still limited. To this end, the present study investigated the growth-inhibitory effects of Lactobacillus casei ATCC 393 against experimental colon cancer. Administration of live Lactobacillus casei (as well as bacterial components thereof) on murine (CT26) and human (HT29) colon carcinoma cell lines raised a significant concentration- and time-dependent anti-proliferative effect, determined by cell viability assays. Specifically, a dramatic decrease in viability of colon cancer cells co-incubated with 10(9) CFU/mL L. casei for 24 hours was detected (78% for HT29 and 52% for CT26 cells). In addition, live L. casei induced apoptotic cell death in both cell lines as revealed by annexin V and propidium iodide staining. The significance of the in vitro anti-proliferative effects was further confirmed in an experimental tumor model. Oral daily administration of 10(9) CFU live L. casei for 13 days significantly inhibited in vivo growth of colon carcinoma cells, resulting in approximately 80% reduction in tumor volume of treated mice. Tumor growth inhibition was accompanied by L. casei-driven up-regulation of the TNF-related apoptosis-inducing ligand TRAIL and down-regulation of Survivin. Taken together, these findings provide evidence for beneficial tumor-inhibitory, anti-proliferative and pro-apoptotic effects driven by this probiotic LAB strain.

Cheese production using kefir culture entrapped in milk proteins.

The aim of the present study was to evaluate the use of kefir culture entrapped in casein and in whey protein as starter cultures for the production of Feta-type cheese. Microbiological analysis showed that counts of enterobacteria, coliforms, and staphylococci were significantly reduced due to kefir culture. In addition, the effect of kefir culture on the formation of volatile compounds, such as esters, organic acids, alcohols, carbonyl compounds, and lactones, was also investigated using the SPME GC/MS technique. Cheese samples produced with kefir culture entrapped in milk proteins presented improved profile of aroma-related compounds. Principal component analysis of the results indicated that the volatile composition of the different cheese types was dependent on the nature of the starter culture. Finally, the sensory evaluation showed that the products produced with kefir culture had a soft, fine taste, and were of improved quality.

Free and immobilized Lactobacillus casei ATCC 393 on whey protein as starter cultures for probiotic Feta-type cheese production.

The use of free and immobilized Lactobacillus casei ATCC 393 on whey protein as starter culture in probiotic Feta-type cheese production was evaluated. The probiotic cultures resulted in significantly higher acidity; lower pH; reduced counts of coliforms, enterobacteria, and staphylococci; and improved quality characteristics compared with cheese with no culture. Microbiological and strain-specific multiplex PCR analysis showed that both free and immobilized L. casei ATCC 393 were detected in the novel products at levels required for conferring a probiotic effect at the end of the ripening. The effect of starter culture on production of volatile compounds was investigated by the solid-phase microextraction gas chromatography-mass spectrometry analysis technique. The immobilized cells resulted in an improved profile of aroma-related compounds and the overall high quality of the novel products was ascertained by the preliminary sensory test. Finally, the high added value produced by exploitation of whey, which is an extremely polluting industrial waste, was highlighted and assessed.

Freeze-dried Saccharomyces cerevisiae cells immobilized on potato pieces for low-temperature winemaking.

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae AXAZ-1 yeast cells on potato pieces. This biocatalyst was subjected to freeze-drying, and the effect of several protective agents and storage at 5 °C, up to 9 months, on viability and fermentative activity of yeasts cells were studied. From several protective agents tested, sodium glutamate preserved the viability of immobilized yeast cells at high levels even after 9-month storage. The freeze-drying biocatalyst was used for repeated batch fermentations of grape must at low temperatures until 5 °C. The produced wines analyzed for volatile byproducts by GC and GC/MS and the results showed that the freeze-dried biocatalysts, with sodium glutamate as protectant, produced wines with higher formation of esters than free cells and having at least similar aromatic profile to those produced by wet biocatalysts.

Monitoring survival of Lactobacillus casei ATCC 393 in probiotic yogurts using an efficient molecular tool.

The aim of the present study was to monitor the survival of the probiotic strain Lactobacillus casei ATCC 393 during refrigerated storage of natural regular yogurts compared with Lactobacillus delbrueckii ssp. bulgaricus. Both free and immobilized cells on supports of high industrial interest, such as fruits and oat pieces, were tested. Microbiological and strain-specific multiplex PCR analysis showed that both free and immobilized Lb. casei ATCC 393 were detected in the novel products at levels required to confer a probiotic effect (at least 6 log cfu/g) for longer periods than required by the dairy industry (≥ 30 d) during storage at 4°C. In contrast, the viable bacterial density of Lb. delbrueckii ssp. bulgaricus decreased to levels <6 log cfu/g after 14 d of cold storage. Of note, the final pH of all products was 4.2 to 4.3. Acid resistance or cold tolerance of Lb. casei ATCC 393 apparently allows for increased survival compared with Lb. delbrueckii ssp. bulgaricus in these yogurt formulations.

Scale-up of thermally dried kefir production as starter culture for hard-type cheese making: an economic evaluation.

This paper concerns the effect of thermal-drying methodology on the investment cost for dried kefir cells production in order to be used as starter culture in cheese manufacturing. Kefir cells were produced at pilot plant scale using a 250-L bioreactor and whey as the main substrate. Kefir cells were subsequently dried in a thermal dryer at 38 degrees C and used as a starter culture in industrial-scale production of hard-type cheeses. The use of thermally dried kefir as starter culture accelerated ripening of cheeses by increasing both lipolysis and fermentation rate as indicated by the ethanol, lactic acid, and glycerol formation. Additionally, it reduced coliforms and enterobacteria as ripening proceeded. This constituted the basis of developing an economic study in which industrial-scale production of thermally dried kefir starter culture is discussed. The industrial design involved a three-step process using three bioreactors of 100, 3,000, and 30,000 L for a plant capacity of 300 kg of thermally dried kefir culture per day. The cost of investment was estimated at 238,000 euro, which is the 46% of the corresponding cost using freeze-drying methodology. Production cost was estimated at 4.9 euro/kg of kefir biomass for a 300-kg/day plant capacity, which is the same as with the corresponding cost of freeze-dried cells. However, the estimated added value is up to 10.8 x 10(9) euro within the European Union.

Whey valorisation: a complete and novel technology development for dairy industry starter culture production.

Whey is the major by-product of the dairy industry, produced in large quantities and usually disposed off causing major environmental pollution, due to its high organic load that makes treatment cost prohibitive. This paper comprises a contribution on the valorisation of this high polluting liquid waste of the dairy industry, based on research for the production of novel dairy starter cultures using whey as raw material. Starter cultures are used for cheese ripening in order to: (i) accelerate ripening, (ii) improve quality and (iii) increase shelf-life. The developed technology involves biomass production from whey followed by thermal drying of cultures. Specifically, Kluyveromyces marxianus, Lactobacillus bulgaricus and kefir yeasts were thermally dried, and their efficiency in lactose and milk whey fermentations was studied. The most suitable culture regarding its technological properties was kefir, which was used for cheese ripening in freeze-dried and thermally dried form. Besides the reduction of production cost, which is an essential requirement for the food industry, the use of thermally dried kefir displayed several other advantages such as acceleration of ripening, increase of shelf-life, and improvement of hard-type cheese quality.