Dry rose extract (DRE) obtained industrially by aqueous ethanol extraction from R. damascena flowers and its phenolic-enriched fraction, obtained by re-extraction with ethyl acetate (EAE) were the subject of this study. 1H NMR of DRE allowed the identification and quantitation of fructose and glucose, while the combined use of HPLC-DAD-ESIMS and HPLC-HRMS showed the presence of 14 kaempferol glycosides, 12 quercetin glycosides, 4 phenolic acids and their esters, 4 galloyl glycosides, 7 ellagitannins, and quinic acid. In addition, the structures of 13 of the flavonoid glycosides were further confirmed by NMR. EAE was found to be richer in TPC and TFC and showed better antioxidant activity (DPPH, ABTS, and FRAP) compared to DRE. Both extracts displayed significant activity against Propionibacterium acnes, Staphylococcus aureus, and S. epidermidis, but showed no activity against Candida albicans. Toxicity tests on normal human skin fibroblasts revealed low toxicity for both extracts with stronger effects observed at 24 hours of treatment that were compensated for over the following two days. Human hepatocarcinoma (HepG2) cells exhibited an opposite response after treatment with a concentration above 350 µg/mL for EAE and 500 µg/mL for DRE, showing increased toxicity after the third day of treatment. Lower concentrations were non-toxic and did not significantly affect the cell cycle parameters of either of the cell lines.
The applicability of two lactic acid bacterial strains with probiotic potential and bioprotective properties as additions in the starter culture in yogurt fermentation was examined. The studied strains, Lactobacillus delbrueckii subsp. bulgaricus KZM 2-11-3 and Lactiplantibacillus plantarum KC 5-12, inhibited the growth of Kluyveromyces lactis, Kluyveromyces marxianus, and Saccharomyces cerevisiae. The strain L. delbrueckii subsp. bulgaricus KZM 2-11-3 directly inhibited Escherichia coli. The important characteristics for the quality of the yogurt product, such as physicochemical parameters during fermentation and storage, rheological characteristics, and sensory changes during the storage of samples were determined. The yogurt samples with the strains did not differ in most parameters from the control yogurt with the commercial starter. The added strains showed stable viability in the yogurt samples during storage. The yogurt sample with L. delbrueckii subsp. bulgaricus KZM 2-11-3 and the sample with both strains based on the total evaluation were very similar to the control yogurt with the commercial starter. Using these strains as probiotic supplements to enrich the starter cultures in yogurt production will contribute to developing new products with benefits to human health.
1 October 2018 in Biocatalysis and Agricultural Biotechnology
Oral probiotic lactic acid bacteria can exhibit antagonistic activities against pathogens associated with diseases in the oral cavity. Therefore, twelve previously isolated oral strains were assessed for antagonistic evaluation against selected oral test microorganisms Streptococcus mutans and Candida albicans. Two separate co-culturing analyses were performed, where all tested strains showed the presence of antagonistic activity and four strains, Limosilactobacillus fermentum N 2, TC 3-11, and NA 2-2, and Weissella confusa NN 1, significantly inhibited Streptococcus mutans by 3–5 logs. The strains showed antagonistic activity against Candida albicans, and all exhibited pathogen inhibition by up to 2 logs. Co-aggregation capability was assessed, showing co-aggregative properties with the selected pathogens. Biofilm formation and antibiofilm activity of the tested strains against the oral pathogens were assayed, where the strains showed specificity in self-biofilm formation and well-expressed antibiofilm properties by most of them above 79% and 50% against Streptococcus mutans and Candida albicans, respectively. The tested LAB strains were assayed by a KMnO4 antioxidant bioassay, where most of the native cell-free supernatants exhibited total antioxidant capacity. These results show that five tested strains are promising candidates to be included in new functional probiotic products for oral healthcare.
The whole-genome sequencing of lactic acid bacteria provides a valuable resource for identifying the genetic determinants underlying molecular mechanisms related to their probiotic properties. Analysis of draft genome sequences relies on bioinformatics tools for genetic data processing and in silico analytical methods to pinpoint the genetic determinants encoding biologically active compounds. The aim of this study was to perform the phenotypic determination of the antibiotic sensitivity and bioinformatics analyses on whole-genome assemblies from LAB from the human oral microbiome, and determine the presence of acquired antibiotic resistance genes, peptidases, adhesion proteins, and bacteriocins. Bioinformatics processing was performed in order to establish the molecular mechanisms responsible for the previously observed probiotic properties. The tested LAB strains exhibited a broad spectrum of antibiotic multiresistance, but did not possess acquired antibiotic resistance genes. The detected genes for peptidase activity were from the Pep family of hydrolase enzymes. Genetic determinants for adhesion proteins contained LPxTG, YSIRK, KxYKxGKxW, and SEC 10/PgrA domains, as well as MucBP domains. Lectins were found for five of the strains with the presence of WxL domains from the CscC protein family and L-type lectin domains. The in silico analyses show that some of the tested strains possessed mechanisms for bacteriocin production.
Lactic acid bacteria are widely studied microorganisms and are one of the prevalent groups of bacteria in the oral cavity microbiome. This work aimed to isolate new lactic acid bacterial strains from the human oral cavity and evaluate their characteristics and probiotic potential. Twelve strains were isolated and identified as belonging to several genera in the family Lactobacillaceae. Screening for antimicrobial activity was held, where two of the strains showed antagonistic activity against Streptococcus mutans and most of the strains expressed inhibition against Escherichia coli, Bacillus subtilis, and Bacillus cereus. The ability of the studied strains to autoaggregate and bind to mucin was assessed, showing autoaggregative properties and mucin binding at 5 logs CFU/mL. The survival ability in simulated oral and gastrointestinal conditions and growth dynamics with different gastrointestinal stress factors was studied. Most of the strains showed a good growth potential in the presence of oral and gastrointestinal stress factors. All tested strains exhibited high survival rates in the simulated oral environment, thus having the potential for colonizing the oral cavity and their beneficial properties to be applied. These results are a good basis for continuing the research into these strains so they can be included in new functional products for oral health.