Diversity and Probiotic Potential of Lactic Acid Bacteria Isolated from Horreh,a Traditional Iranian Fermented Food

The aim of this study was to evaluate the probiotic potential of lactic acid bacteria (LAB) strains isolated from Horreh. Some probiotic properties, e.g., resistance to acid, bile tolerance, antibacterial activity, and antibiotic susceptibility, were investigated. A total of 140 Gram-positive and catalase-negative isolates from Horreh were subjected to identification and grouping by cultural methods and the 16S rRNA sequencing. The new isolates were identified to be Lactobacillus (fermentum, plantarum, and brevis) Weissella cibaria, Enterococcus (faecium and faecalis), Leuconostoc (citreum and mesenteroides subsp. mesenteroides) and Pediococcus pentosaceus. Probiotic potential study of LAB isolates showed that Lb. plantarum and Leu. mesenteroides subsp. mesenteroides isolates were able to grow at pH 2.5 and 3.5. Lactobacillus plantarum (isolate A44) showed the highest cell hydrophobicity (84.5%). According to antibacterial activity tests, Listeria innocua and Staphylococcus aureus were the most sensitive indicators against the selected LAB strains, while Escherichia coli and Bacillus cereus were the most resistant. In addition, all the isolated LAB species were resistant to vancomycin. The results of the present study suggested that the Lactobacillus fermentum and plantarum isolated from Horreh, characterized in this study, have potential use for industrial purposes as probiotics.     

Probiotic Potential of Lactobacillus plantarum LD1 Isolated from Batter of Dosa,a South Indian Fermented Food

Lactobacillus plantarum LD1 was isolated from dosa batter and identified by biochemical, physiological and genetic methods. Species level identification was done by 16S rDNA amplification and sequencing. The probiotic potential of strain LD1 was assessed by different standard parameters. Cell surface hydrophobicity was recorded to be 62 % with SAT value <0.007 M. Seventy-eight percent of viable count was found after treatment with simulated gastric juice containing pepsin (pH 2.0). Bile salt tolerance and bile salt hydrolase activity were also demonstrated by strain LD1. The culture supernatant was able to inhibit food-borne as well as clinical pathogenic microorganisms such as Staphylococcus aureus, Salmonella typhimurium, Shigella flexneri, Pseudomonas aeruginosa, urogenic Escherichia coli and Vibrio sp. Strain LD1 was found to be sensitive to most of the antibiotics used in the study. Since strain has been isolated from food source that is most typical of Southern India, it would be safe for further consumption in probiotic products.     

In Vitro Assessment of Probiotic Potential of Saccharomyces Cerevisiae DABRP5 Isolated from Bollo Batter,a Traditional Goan Fermented Food

Bollo is a traditional Goan fermented food in which coarse wheat/wheat and finger millet is leavened with toddy. We here isolated 42 yeast strains from Bollo batter. Initial screening of the isolates with probiotic properties yielded four yeast isolates (DABRP1, DABRP2, DABRP5 and DABRP12). These isolates exhibited tolerance to high bile salt concentration and acidic pH and resistance to various antibiotics, which indicated their probiotic nature. All these yeast isolates were identified as Saccharomyces cerevisiae through D1D2-LSU-rDNA sequencing. These yeast isolates also showed higher percent hydrophobicity towards chloroform followed by n-hexadecane and o-xylene indicating their mucosal surface-adhesive property. To evaluate the safety of the isolates for them to be called as generally recognized as safe, the pathogenic behavior of the isolates determined through the in vitro hemolysis assay and evaluation of DNase and gelatinase activities. None of the isolates exhibited hemolysis or produced DNase or gelatinase and thus were considered potentially safe. In terms of beneficial effects, the most potent isolate S. cerevisiae DABRP5 showed antibacterial activity against the test pathogens. It also showed excellent antioxidant activity with DPPH free radical scavenging activity of 68.85 ± 0.69%, anti-inflammatory activity with 60.39 ± 0.34% inhibition of protein denaturation, and antidiabetic activity with 71.75 ± 0.45% inhibition of α-amylase activity. The isolate produced α-amylase, lipase, and β-galactosidase. The probiotic potential of the isolate S. cerevisiae DABRP5 was similar to that of the reference strain (Saccharomyces boulardii CNCM I-745) used in this study. The results thus indicate that yeast isolates from Bollo batter have probiotic potential.

     

Evaluation of Probiotic Potential of Bacteriocinogenic Lactic Acid Bacteria Strains Isolated from Meat Products

In this study, the probiotic potential of five bacteriocin-producing lactic acid bacteria (LAB) strains, isolated from meat products, was investigated. They were presumptively identified as Lactococcus lactis subsp. cremoris CTC 204 and CTC 483, L. lactis subsp. hordinae CTC 484, and Lactobacillus plantarum CTC 368 and CTC 469 according to morphological, biochemical, and physiological characteristics. Analysis of genetic variability (random amplified polymorphic (RAPD)-PCR) and whole-cell proteins (SDS-PAGE) revealed similarity between Lactobacillus strains and variability among Lactococcus strains. The evaluation of the probiotic potential showed that the five LAB strains were tolerant to pH 2.0, and only strain CTC 469 was tolerant to the lowest concentration of the bile salts evaluated (0.1%). All strains showed survival or growth ability at 4, 25, and 37 °C, and tolerance at ??20 °C. Although strain CTC 204 in TSB Broth supplemented with MgSO₄ showed the highest intensity of biofilm production, this compound was produced by all of them. The safety assessment showed that no thermonuclease, hemolytic, or gelatinase activities were detected. All strains were resistant to erythromycin and sensitive to amoxicillin and phenoxymethylpenicillin; furthermore, CTC 204 was resistant to chloramphenicol, CTC 368 and CTC 469 to chloramphenicol and vancomycin, CTC 483 to tetracycline and vancomycin, and CTC 484 to clindamycin and chloramphenicol. The evaluated strains showed biogenic amine production; the lowest levels were produced by CTC 204 and CTC 368 strains. It was concluded that CTC 204 and CTC 368 strains have the greatest potential for becoming probiotics.     

Phenotypic and Genotypic Identification of the Most Acidifiers LAB Strains Isolated from Fermented Food

Biology Bulletin - Over 18 strains of lactic acid bacteria “LAB” isolated from different traditionally fermented products mainly sourdough and fermented vegetables, have been...     

Bacteriocinogenic Potential of Bacillus amyloliquefaciens Isolated from Kimchi,a Traditional Korean Fermented Cabbage

Bacteriocin production is considered a favorable property for various beneficial cultures. In addition to their potential as biopreservatives, bacteriocins are also promising alternatives for the control of multidrug-resistant pathogens and the inhibition of some viruses and cancer cells. The objective of this study was to screen and characterize a bacteriocin-producing strain with the aim of its future application for control of Listeria monocytogenes, an important food-borne pathogen. A total of 22 potentially bacteriocinogenic strains active against L. monocytogenes ATCC15313 were isolated from locally produced kimchi through a three-level approach. Pure cultures were obtained according to good microbiological practices and differentiated through RAPD-PCR using the primers OPL01, OPL09, and OPL11. Altogether, 5 strains were selected for further study. Specific focus was given to strain ST05DL based on its specific inhibitory activity against L. monocytogenes ATCC15313, while not affecting different strains belonging to the genera Lactobacillus, Pediococcus, Leuconostoc, and Weissella, most of which are beneficial microorganisms. The strain ST05DL was identified as Bacillus amyloliquefaciens based on its sugar fermentation profile obtained through API50CHB analysis and 16S rRNA partial sequencing. The antimicrobial compound produced by B. amyloliquefaciens ST05DL was found to be sensitive to pepsin and α-chymotrypsin, evidence of its proteinaceous nature. The presence of skim milk, NaCl, Tween 80, glycerol, and SDS did not affect the antimicrobial activity. The addition of 20% cell-free supernatant (CFS) obtained from a 24-h culture of B. amyloliquefaciens ST05DL to an exponentially growing culture of L. monocytogenes ATCC15313 successfully inhibited the test microorganisms during the monitored 10-h incubation. Optimal bacteriocin production by B. amyloliquefaciens ST05DL was observed during the stationary phase at 12 h (800 AU/mL) and remained stable for the next 15 h. The ratio between live and dead cells during this period was 74.37% and 25.66%, respectively, as determined by flow cytometry. The presence of the virulence genes hblA, hblB, hblC, nheA, nheB, and nheC was not detected in the total DNA of B. amyloliquefaciens ST05DL, and the strain was resistant only to ampicillin out of 10 tested antibiotics. Future evaluation of expressed bacteriocin/s by B. amyloliquefaciens ST05DL (amino acid sequence, molecular mass, cytotoxicity, detailed mode of action, etc.), will be the next step in the characterization and its potential application as biopreservative and/or pharmaceutical product.

     

Probiotic Potential,Antioxidant Activity,and Phytase Production of Indigenous Yeasts Isolated from Indigenous Fermented Foods

While many bacteria have been used as probiotics by industries, only two yeasts, Saccharomyces cerevisiae var. boulardii and Kluyveromyces fragilis (B0399), have been used for this purpose. In the present work, a total of 116 yeasts isolated from Brazilian indigenous fermented food, cocoa fermentation, and kefir were in vitro characterized for probiotic attributes. From 116 isolates, 36 were tolerant to gastrointestinal conditions evaluated by tolerance to pH 2.0, bile salts (0.3% w/v), and 37 °C temperature. From those, 15 isolates showed a similar or higher percentage (P < 0.05) of hydrophobicity, autoaggregation, and coaggregation with E. coli than the reference strain S. boulardii. All these strains showed a high percentage of adhesion to Caco-2 cells (> 63%) and antioxidant activity (ranging from 18 to 62%). Phytate hydrolysis was evaluated for these yeasts and 13 strains showed positive results, which is important for nutrient availability in plant-based foods. These results are important insights for characterization of novel probiotic yeast strains as well as to aggregate functional value to these food products.

     

Probiotic Properties of Lactobacillus Strains Isolated from Tibetan Kefir Grains

The objective of this study was to evaluate the functional properties of lactic acid bacteria (LAB) isolated from Tibetan kefir grains. Three Lactobacillus isolates identified as Lactobacillus acidophilus LA15, Lactobacillus plantarum B23 and Lactobacillus kefiri D17 that showed resistance to acid and bile salts were selected for further evaluation of their probiotic properties. The 3 selected strains expressed high in vitro adherence to Caco-2 cells. They were sensitive to gentamicin, erythromycin and chloramphenicol and resistant to vancomycin with MIC values of 26 µg/ml. All 3 strains showed potential bile salt hydrolase (BSH) activity, cholesterol assimilation and cholesterol co-precipitation ability. Additionally, the potential effect of these strains on plasma cholesterol levels was evaluated in Sprague-Dawley (SD) rats. Rats in 4 treatment groups were fed the following experimental diets for 4 weeks: a high-cholesterol diet, a high-cholesterol diet plus LA15, a high-cholesterol diet plus B23 or a high-cholesterol diet plus D17. The total cholesterol, triglyceride and low-density lipoprotein cholesterol levels in the serum were significantly (P<0.05) decreased in the LAB-treated rats compared with rats fed a high-cholesterol diet without LAB supplementation. The high-density lipoprotein cholesterol levels in groups B23 and D17 were significantly (P<0.05) higher than those in the control and LA15 groups. Additionally, both fecal cholesterol and bile acid levels were significantly (P<0.05) increased after LAB administration. Fecal lactobacilli counts were significantly (P<0.05) higher in the LAB treatment groups than in the control groups. Furthermore, the 3 strains were detected in the rat small intestine, colon and feces during the feeding trial. The bacteria levels remained high even after the LAB administration had been stopped for 2 weeks. These results suggest that these strains may be used in the future as probiotic starter cultures for manufacturing novel fermented foods.     

Whole-Genome Sequencing and Comparative Genome Analysis of Bacillus subtilis Strains Isolated from Non-Salted Fermented Soybean Foods

Bacillus subtilis is the main component in the fermentation of soybeans. To investigate the genetics of the soybean-fermenting B. subtilis strains and its relationship with the productivity of extracellular poly-γ-glutamic acid (γPGA), we sequenced the whole genome of eight B. subtilis stains isolated from non-salted fermented soybean foods in Southeast Asia. Assembled nucleotide sequences were compared with those of a natto (fermented soybean food) starter strain B. subtilis BEST195 and the laboratory standard strain B. subtilis 168 that is incapable of γPGA production. Detected variants were investigated in terms of insertion sequences, biotin synthesis, production of subtilisin NAT, and regulatory genes for γPGA synthesis, which were related to fermentation process. Comparing genome sequences, we found that the strains that produce γPGA have a deletion in a protein that constitutes the flagellar basal body, and this deletion was not found in the non-producing strains. We further identified diversity in variants of the bio operon, which is responsible for the biotin auxotrophism of the natto starter strains. Phylogenetic analysis using multilocus sequencing typing revealed that the B. subtilis strains isolated from the non-salted fermented soybeans were not clustered together, while the natto-fermenting strains were tightly clustered; this analysis also suggested that the strain isolated from “Tua Nao” of Thailand traces a different evolutionary process from other strains.     

鸡源益生芽胞杆菌的筛选

根据菌体对低pH和高胆盐的耐受性、产酶特性和对病原菌的抑菌效果,对家鸡肠道中的芽胞杆菌进行了筛选。结果表明,有4株芽胞杆菌(A2、C5、C19和D8)能抑制金黄色葡萄球菌(Staphylococcus aureus)的生长。这4株菌在pH 3.0和高胆盐条件下的存活率都达到60%以上,产淀粉酶和蛋白酶的能力也较好,表明这些菌株具有作为益生菌的开发潜力。经形态学和生理生化反应鉴定,初步确定A2和D8为地衣芽胞杆菌(Bacillus licheniform is),C5和C19为蜡样芽胞杆菌(Bacillus cereus)。     

Probiotic Properties of Lactobacillus Species Isolated from Fermented Palm Sap in Thailand

Probiotics and Antimicrobial Proteins - Lactic acid bacteria (LAB), which are the most frequently used probiotics in foods, confer health benefits such as antimicrobial activity, immune...     

New Potentially Probiotic Strains Isolated from Humans – Comparison of Properties with Strains from Probiotic Products and ATCC Collection

Lactic acid bacteria are used in various types of probiotic products. Due to the constantly growing probiotics market, new strains with pro-health properties are sought. The present study compared 39 strains of Lactobacillus, Lacticaseibacillus, and Lactiplantibacillus, isolated from probiotic products and healthy people. The current research aimed to search for new, potentially probiotic strains. For this purpose the relationship between Lactobacillaceae strains was carried out; moreover, the basic properties of probiotic microorganisms, such as survival at low pH and bile salt environment, antibiotic susceptibility, aggregation and antagonism were estimated. The properties of these isolates were also compared with the properties of probiotic strains from the ATCC collection. In comparing the genetic relationship (PFGE method) between the tested isolates, it was observed that some of them show a high degree of similarity. All tested strains tolerated an environment with a pH value of 3.0, and the addition of 0.3% bile salt; showed auto-aggregation properties and displayed antagonism against pathogenic microorganisms. In the present study, the bacteria were susceptible to tetracycline, chloramphenicol and ampicillin; the resistance to vancomycindepended on the bacteria type. All the properties were strain-depended. Most of the tested strains had properties comparable to the reference strains. Three L. acidophilus strains isolated from cervical swabs seem to be promising candidates for probiotic strains. Key words: lactobacilli, PFGE, antimicrobial susceptibility, antagonism     

Antibacterial Peptides, Probiotic Properties and Biopreservative Efficacy of Native Bacillus Species Isolated from Different Food Sources

In this study, we evaluated the occurrence of antibacterial peptide (ABP)-producing Bacillus spp. in fermented foods. Among 78 isolated cultures, 25 potential ABP-producing stains were selected and differentiated genotypically and phenotypically. The 16S rRNA gene sequence homology, in combination with morphological, physiological and biochemical characteristics, was used for the identification of the isolates. The isolates exhibited inhibitory activity against both Gram-positive and Gram-negative food-borne pathogens. The antibacterial compounds produced by these cultures were proteinaceous in nature, with molecular weight falling in the range of 3?C6.5?kDa. The ABP present in the cell-free supernatant of B. subtilis Ec1 and B. licheniformis Me1 exhibited the highest titre of activity (3,400?AU/ml) and wide range of pH (4?C10) and temperature (40?C100?°C) stability. The strain Ec1 was found to be exhibiting some in vitro probiotic properties, such as acid and bile tolerance, bile salt hydrolase activity and hydrophobicity towards hydrocarbons. The viable counts of Listeria monocytogenes Scott A in pasteurized milk samples containing ABP of Ec1 were lower than those observed in controls without ABP. The ABP-coated packaging films exhibited antimicrobial activity against the pathogens, indicating the application of ABP from Bacillus spp. in antimicrobial packing materials. These observations increase the likelihood of potential use of the isolated Bacillus spp. or their ABP for application in food biopreservation and as probiotics.     

Aggregation and Adhesion Activity of Lactobacilli Isolated from Fermented Products In Vitro and In Vivo: a Potential Probiotic Strain

Approximately 25 strains of lactobacilli isolated from different dairy products and fermented vegetables were screened according to their possibility to show the high auto-aggregation and co-aggregation. The strains Lactobacillus helveticus INRA-2010-H11, Lactobacillus rhamnosus INA-5.1, and Lactobacillus acidophilus JM-2012 were determined to have the high auto-aggregation (approximately 73, 46, and 70.5% correspondingly). A high co-aggregation capacity (75.53%) for strains INRA-2010-H11 and JM-2012 was shown. The adhesion degree of INRA-2010-H11 on the surface of buccal epithelial cells was 88.23%. The study of INRA-2010-H11, JM-2012, and both strains’ mixture (1:1) adhesion capacity on the surface of epithelial HeLa cells revealed the adhesion of 1.1 × 10⁶, 6.3 × 10⁴, and 2.3 × 10⁵ CFU, respectively, from starter amount of CFU 10⁷ and 10⁸ for both strains. In vivo experiments of LAB adhesion in gastrointestinal tract of mouse revealed the presence of 2.5 × 10⁹, 1.2 × 10⁹, and 1.5 × 10⁹ CFU of LAB in control and groups of mouse, fed by INRA-2010-H11 and mixture, respectively. Feeding by investigated lactobacilli was suggested to lead to microbiota biodiversity reduction in small intestine and colon and its augmentation in stomach. Thus, INRA-2010-H11 demonstrated a high aggregation and adhesion activity so it has the potential as a good probiotic strain.     

In Vitro Characterization of Lactic Acid Bacteria Isolated from Bovine Milk as Potential Probiotic Strains to Prevent Bovine Mastitis

Probiotics and Antimicrobial Proteins - Bovine mastitis causes economic losses on dairy farms worldwide. Lactic acid bacteria (LAB) in animal health are an alternative tool to avoid antibiotic...     

Craft Beers Fermented by Potential Probiotic Yeast or Lacticaseibacilli Strains Promote Antidepressant-Like Behavior in Swiss Webster Mice

Probiotics and Antimicrobial Proteins - This study aimed to produce a probiotic-containing functional wheat beer (PWB) by an axenic culture system with potential probiotic Saccharomyces cerevisiae...     

Pathogenic Potential of Saccharomyces Strains Isolated from Dietary Supplements

Saccharomyces cerevisiae plays a beneficial role in health because of its intrinsic nutritional value and bio-functional properties, which is why it is also used as a dietary supplement. However, the perception that S. cerevisiae is harmless has changed due to an increasing number of infections caused by this yeast. Given this scenario, we have tested whether viable strains contained in dietary supplements displayed virulence-associated phenotypic traits that could contribute to virulence in humans. We have also performed an in vivo study of the pathogenic potential of these strains using a murine model of systemic infection by intravenous inoculation. A total of 5 strains were isolated from 22 commercial products and tested. Results highlight one strain (D14) in terms of burden levels in brains and kidneys and ability to cause death, whereas the other two strains (D2 and D4) were considered of low virulence. Our results suggest a strong relationship between some of the virulence-associated phenotypic traits (ability to grow at 39°C and pseudohyphal growth) and the in vivo virulence in a mouse model of intravenous inoculation for isolates under study. The isolate displaying greatest virulence (D14) was evaluated in an experimental murine model of gastrointestinal infection with immunosuppression and disruption of mucosal integrity, which are common risk factors for developing infection in humans, and results were compared with an avirulent strain (D23). We showed that D14 was able to spread to mesenteric nodes and distant organs under these conditions. Given the widespread consumption of dietary supplements, we recommend only safe strains be used.     

Role of Leuconostoc mesenteroides in Leavening the Batter of Idli, a Fermented Food of India

The fermentation of the batter of idli, a fermented food of India, was studied. The microorganisms responsible for the characteristic changes in the batter were isolated and identified. Although there is a sequential change in the bacterial flora, the predominant microorganism responsible for souring, as well as for gas production, was found to be Leuconostoc mesenteroides. In the later stages of fermentation, growth of Streptococcus faecalis and, still later, of Pediococcus cerevisiae becomes significant. The fermentation of idli demonstrates a leavening action caused by the activity of the heterofermentative lactic acid bacterium, L. mesenteroides. As far as is known, this is the first record of a leavening action produced exclusively by the activity of a lactic acid bacterium.     

Comparative Studies of Bacillus Cereus Strains Isolated From Various Foods and Food Poisoning Outbreaks

Certain properties of 22 Bacillus cereus strains isolated from different foods and food poisoning episodes were investigated in order to evaluate possible différences between strains isolated from diarrhoeal and vomiting type food poisoning outbreaks. None of the strains isolated from vomiting type episodes produced acid from salicin and mannose, whereas 80 and 40 % of the strains from diarrhoeal type outbreaks were positive, respectively. No association between the antibiotic sensitivity pattern or the fatty acid composition and the source of a strain could be found, although some strains differed from the general pattern of B. cereus in some instances. No significant differences in the production of the skin factor between strains isolated from the two types of outbreaks were found either. The findings of this study support the observation that the food environment itself essentially affects the enterotoxin formation of B. cereus.