Biofunctional Attributes and Storage Study of Soy Milk Fermented by Lactobacillus rhamnosus and Lactobacillus helveticus.

Different soybean products are considered as traditional functional food among the Mongolian population in Northeast India. In the present study, the effect of different flavours (mango, orange, vanilla and white rose), inoculation rates of Lactobacillus rhamnosus K4E (KX950834) and Lactobacillus helveticus K14 (KU644578), and mass fractions of skimmed milk and sugar on the acceptability of soy yoghurts was studied. Physicochemical (pH, titratable acidity) and microbial analyses (total bacterial, total coliform, yeast and mould count) were conducted, and organoleptic (aroma, taste, colour, mouthfeel, texture and overall acceptability) and biofunctional properties (angiotensin-converting enzyme (ACE) inhibitory activity, antioxidant and antimicrobial activities and biotransformation of isoflavones) were evaluated during storage up to 10 days under refrigeration conditions (6-8 °C). Panellists preferred white rose soy yoghurt more than other flavours. The pH was from 5.65 to 4.20, the titratable acidity (expressed as mass fraction of lactic acid) was from 0.33 to 0.51% and total Lactobacillus count ranged from 6.81 to 8.69 log CFU/mL during storage. The ACE inhibitory activity increased from 21.17% on day 0 to 81.03% on day 5, followed by a decrease of the activity after 10 days (38.85%). The antioxidant activity was the highest on day 5 (87%). White rose soy yoghurt had the highest antimicrobial activity against Listeria monocytogenes, followed by Bacillus subtilis, Staphylococcus aureus, Salmonella typhi and Escherichia coli. RP-HPLC analysis showed that after 18 h, the production of soy isoflavone aglycones genistein and daidzein in yoghurt was 87.3 and 58.4%, respectively.

Whole-genome based validation of the adaptive properties of Indian origin probiotic Lactobacillus helveticus MTCC 5463.

BACKGROUND: The aim of the study was to mine the Lactobacillus helveticus MTCC 5463 genome for genetic determinants to validate its ability to adapt to gut transit stresses and translate functionality to the host. RESULTS: In silico analysis of the 1 911 350 bp single chromosome of the strain predicted that it had excellent adaptive features like the multisubunit F0F1 ATPase, conjugated bile salt hyrolase, chaperones like hsp33, HtrA, GroEL, GroES, dnaK, grpE, starvation-inducible proteins and heavy-metal transporting ATPases. The genome revealed genes for adhesion and aggregation including exopolysaccharides, capsular polysaccharides sortase, elongation factor Tu, aggregation promoting proteins, fibronectin-binding proteins, S-layer and mucus-binding proteins. We could identify genes conferring physiological benefits like immunostimulation, cholesterol reduction, antibacterial and folate production. Thus, through trait and gene matching, the study established that the strain possessed the genetic arsenal required to adapt to the gut milieu. The predictions of functional genes further validate the experimental evidences of adaptation and probiosis. CONCLUSION: This study provides insight into the feasibility of applying probiogenomics to identify genes that could function as pre-selection criteria for identification of potential probiotic strains.

Comparative genome-scale analysis of niche-based stress-responsive genes in Lactobacillus helveticus strains.

Next generation sequencing technologies with advanced bioinformatic tools present a unique opportunity to compare genomes from diverse niches. The identification of niche-specific stress-responsive genes can help in characterizing robust strains for multiple applications. In this study, we attempted to compare the stress-responsive genes of a potential probiotic strain, Lactobacillus helveticus MTCC 5463, and a cheese starter strain, Lactobacillus helveticus DPC 4571, from a gut and dairy niche, respectively. Sequencing of MTCC 5463 was done using 454 GS FLX, and contigs were assembled using GS Assembler software. Genome analysis was done using BLAST hits and the prokaryotic annotation server RAST. The MTCC 5463 genome carried multiple orthologs of genes governing stress responses, whereas the DPC 4571 genome lacked in the number of major stress-response proteins. The absence of the bile salt hydrolase gene in DPC 4571 and its presence in MTCC 5463 clearly indicated niche adaptation. Further, MTCC 5463 carried higher copy numbers of genes contributing towards heat, cold, osmotic, and oxidative stress resistance as compared with DPC 4571. Through comparative genomics, we could thus identify stress-responsive gene sets required to adapt to gut and dairy niches.

Hypocholesterolemic Effect of Potential Probiotic Lactobacillus fermentum Strains Isolated from Traditional Fermented Foods in Wistar Rats.

The current research project was undertaken to explore the therapeutic potential of two potent probiotic Lactobacillus fermentum strains, i.e., PD2 and PH5 in a hyperlipemic healthy adult Wistar rat model, with a particular focus as biotherapeutics for the management of high cholesterol in Indian population. Rats fed on cholesterol-enriched diet supplemented with potential probiotics strain Lactobacillus fermentum PH5 significantly affected serum lipid profile by reducing serum cholesterol (67.21%), triglycerides level (66.21%), and LDL cholesterol level (63.25%) in comparison to rats that received cholesterol-enriched diet (Model) only. Both the strains decreased the cholesterol levels in liver compared with Model group, but PH5 was found to be more effective (30.65% reduction) in liver total cholesterol (TC) lowering action. In addition, the fecal coliforms were significantly reduced besides increased LAB in feces of rats receiving probiotic curd having Lactobacillus fermentum PH5. Our results demonstrated that supplementation with either of the two strains was efficient in reducing serum cholesterol, LDL-cholesterol and TG concentrations in rats compared to those fed the same high-cholesterol diet but without LAB supplementation.

Complete Genome Sequence of Lactobacillus fermentum MTCC 25067 (Formerly TDS030603), a Viscous Exopolysaccharide-Producing Strain Isolated from Indian Fermented Milk.

Lactobacillus fermentum MTCC 25067 (formerly TDS030603) is capable of producing a highly viscous slime exopolysaccharide. We report here the complete genome sequence of the strain, which was deciphered by using PacBio single-molecule real-time sequencing technology.

Effect of Probiotic Dietary Intervention on Calcium and Haematological Parameters in Geriatrics.

INTRODUCTION: Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. Sufficient calcium intake has been reported to support bone growth and prevent bone loss during the ageing process. AIM: To determine the effect of Lactobacillus helveticus MTCC 5463 probiotic dietary intervention on serum calcium & haematological parameters in geriatric population. MATERIALS AND METHODS: Healthy volunteers with age ranging from 64-74 years were recruited from the nearby residential areas in and around Anand, Gujarat. Study duration was from 2012 to 2015. Of the 112 subjects initially enrolled in the trial, 36 withdrew before the intervention because of not matching with criterias. Of the 76 participants, 5 subjects (4%) under test group and 12 subjects (11%) under placebo left the study. We had 59 subjects who successfully completed a double blind cross over trial. Probiotic fermented milk products (in form of "Lassi") was prepared by supplementing toned milk with honey and fermenting with probiotic Lactobacillus helveticus MTCC 5463 and Streptococcus thermophilus MTCC 5460. The final product had at least 10(8) CFU/ml of viable Lactobacillus helveticus MTCC 5463 at the time of feeding. During feeding period, 200 ml of fermented product containing the test strain to one group and a similar product but without the test strain as placebo were fed regularly at the time of breakfast in morning for 4 weeks. Subjects of each group were given a washout period of 4 weeks before they were crossed over and included to the other group. The study was approved by institutional ethics committee. RESULTS: The socio-demographic and clinical profiles were similar at baseline. The mean (SD) calcium level improved significantly in test {9.36 (0.45) vs 8.45 (0.61), p<0.001}. No significant effect was observed with respect to haemoglobin & haematological parameters. CONCLUSION: The well-documented probiotic Lactobacillus helveticus MTCC 5463 confirmed increase in serum calcium level but no effect on haematological parameters when administered to geriatrics.

Geriatric Respondents and Non-Respondents to Probiotic Intervention Can be Differentiated by Inherent Gut Microbiome Composition.

SCOPE: Probiotic interventions are known to have been shown to influence the composition of the intestinal microbiota in geriatrics. The growing concern is the apparent variation in response to identical strain dosage among human volunteers. One factor that governs this variation is the host gut microbiome. In this study, we attempted to define a core gut metagenome, which could act as a predisposition signature marker of inherent bacterial community that can help predict the success of a probiotic intervention. METHODS AND RESULTS: To characterize the geriatric gut microbiome, we designed primers targeting the 16S rRNA hypervariable region V2-V3 followed by semiconductor sequencing using Ion Torrent PGM. Among respondents and non-respondents, the chief genera of phylum Firmicutes that showed significant differences are Lactobacillus, Clostridium, Eubacterium, and Blautia (q < 0.002), while in the genera of phylum Proteobacteria included Shigella, Escherichia, Burkholderia and Camphylobacter (q < 0.002). CONCLUSION: We have identified potential microbial biomarkers and taxonomic patterns that correlate with a positive response to probiotic intervention in geriatric volunteers. Future work with larger cohorts of geriatrics with diverse dietary influences could reveal the potential of the signature patterns of microbiota for personalized nutrition.

Probiotic Formulations: Application and Status as Pharmaceuticals-A Review.

The use of probiotics is a new way to control and treat infections in this modern era. Application of beneficial bacteria to protect against detrimental bacteria in the gastrointestinal tract and thus reap a positive health benefit is the basis of probiotic therapy. Probiotics have a long global history of traditional use. They are normally consumed through fermented foods and are currently sold mostly as ingredients in foods or nutritional supplements. They are also supplied as pharma products. Recent research has highlighted the probiotic potential in the treatment or prevention of disease conditions, maintenance of health, improving immunity and in the reduction in the risk of future diseases. But their position in the pharmaceutical industry is still not very clear. Clinical practitioners use probiotic pharma products mostly as supplements. Their status as drugs is still unclear. This review is aimed to analyze probiotics as pharmaceuticals, their current status as dietary supplements and drugs, existing probiotic preparations and future research needs.

Genomic analysis of dairy starter culture Streptococcus thermophilus MTCC 5461.

The lactic acid bacterium Streptococcus thermophilus is widely used as a starter culture for the production of dairy products. Whole-genome sequencing is expected to utilize the genetic basis behind the metabolic functioning of lactic acid bacterium (LAB), for development of their use in biotechnological and probiotic applications. We sequenced the whole genome of Streptococcus thermophilus MTCC 5461, the strain isolated from a curd source, by 454 GS-FLX titanium and Ion Torrent PGM. We performed comparative genome analysis using the local BLAST and RDP for 16S rDNA comparison and by the RAST server for functional comparison against the published genome sequence of Streptococcus thermophilus CNRZ 1066. The whole genome size of S. thermophilus MTCC 5461 is of 1.73Mb size with a GC content of 39.3%. Streptococcal virulence-related genes are either inactivated or absent in the strain. The genome possesses coding sequences for features important for a probiotic organism such as adhesion, acid tolerance, bacteriocin production, and lactose utilization, which was found to be conserved among the strains MTCC 5461 and CNRZ 1066. Biochemical analysis revealed the utilization of 17 sugars by the bacterium, where the presence of genes encoding enzymes involved in metabolism for 16 of these 17 sugars were confirmed in the genome. This study supports the facts that the strain MTCC 5461 is nonpathogenic and harbors essential features that can be exploited for its probiotic potential.

Evidence for xylooligosaccharide utilization in Weissella strains isolated from Indian fermented foods and vegetables.

Six strains isolated from fermented food were identified as Weissella species by 16S rDNA sequencing, clustering with the species pair W. confusa/W. cibaria. The strains were analysed for growth on glucose, xylose and xylooligosaccharides (XOS). All strains were xylose positive using the API CHL 50 test. Growth on XOS was observed for strains 85, 92, 145 and AV1, firstly by optical density measurements in microtitre plates and secondly in batch cultures also confirming concomitant decrease in pH. Analysis of XOS before and after growth established consumption in the DP2-DP5 range in the four XOS-fermenting strains. XOS were consumed simultaneously with glucose, while xylose was consumed after glucose depletion. Cell-associated ß-xylosidase activity was detected in the XOS-fermenting strains. Analysis of genomic data suggests this activity to be linked with genes encoding glycoside hydrolases from family 3, 8 or 43. No endo-ß-xylanase activity was detectable. Major end products were lactate and acetate. A higher ratio of acetic acid to lactic acid was obtained during growth on XOS compared with growth on glucose. This is the first report on utilization of XOS in Weissella, indicating an increased probiotic potential for XOS-utilizing strains from the species pair W. confusa/W. cibaria, but also showing that XOS utilization is strain dependent for these species.

Food fermentations: microorganisms with technological beneficial use.

Microbial food cultures have directly or indirectly come under various regulatory frameworks in the course of the last decades. Several of those regulatory frameworks put emphasis on "the history of use", "traditional food", or "general recognition of safety". Authoritative lists of microorganisms with a documented use in food have therefore come into high demand. One such list was published in 2002 as a result of a joint project between the International Dairy Federation (IDF) and the European Food and Feed Cultures Association (EFFCA). The "2002 IDF inventory" has become a de facto reference for food cultures in practical use. However, as the focus mainly was on commercially available dairy cultures, there was an unmet need for a list with a wider scope. We present an updated inventory of microorganisms used in food fermentations covering a wide range of food matrices (dairy, meat, fish, vegetables, legumes, cereals, beverages, and vinegar). We have also reviewed and updated the taxonomy of the microorganisms used in food fermentations in order to bring the taxonomy in agreement with the current standing in nomenclature.