The aim of this study is to characterize the bioactivity of mycelial biomass and crude exopolysaccharides (EPS) produced by Trametes versicolor NBIMCC 8939 and to reveal its nutraceutical potential. The EPS (1.58 g/L) were isolated from a culture broth. The macrofungal biomass was rich in protein, insoluble dietary fibers and glucans. The amino acid composition of the biomass was analyzed and 18 amino acids were detected. Three mycelial biomass extracts were prepared and the highest total polyphenol content (16.11 ± 0.14 mg GAE/g DW) and the total flavonoid content (5.15 ± 0.03 mg QE/g DW) were found in the water extract. The results indicated that the obtained EPS were heteropolysaccharides with glucose as the main building monosaccharide and minor amounts of mannose, xylose, galactose, fucose and glucuronic acid. Fourier Transform Infrared Spectroscopy (FTIR) confirmed the complex structure of the crude EPS. Five probiotic lactic acid bacteria strains were used for the determination of the prebiotic effect of the crude EPS. The anti-inflammatory potential was tested in vitro using cell line HT-29. The significant decrease of IL-1 and IL-8 and increase of TGF-beta expression revealed anti-inflammatory potential of the crude exopolysaccharides from T. versicolor.

The aim of the present study was to analyse the type of adhesive factors of selected probiotic strains. A large number of lactic acid bacteria with intestinal and dairy origin were collected and assessed for adhesion on Caco-2 cell line. From the best adherent bacteria, four strains were selected for further research: Lactobacillus gasseri G7, L. plantarum F1, L. helveticus AC and L. delbrueckii subsp. bulgaricus B14. The average number of adhered bacteria was 17 per one Caco-2 cell in the case of L. gasseri G7 and 21 per cell in the case of L. plantarum F1. Treatment with ethylenediaminetetraacetic acid (EDTA), trypsin and metaperiodic acid in separate assays revealed that cell-bonded extracellular proteins were responsible for the adhesion of the selected L. gasseri, L. plantarum and L. helveticus strains, in contrast to the L. delbrueckii subsp. bulgaricus strain, whose adhesive factors were identified as cell-bonded exopolysaccharides. The cell-wall proteins from the first three strains were isolated, fractionated and assessed for adhesion to Caco-2 cells. Based on the attachment properties of the purified proteins towards Caco-2 cells, it was clearly proved exactly which proteins are involved in the adherence. L. plantarum F1 strain contains two adhesive proteins in contrast to the other selected strains containing one adhesive protein each. The determination of the factors mediating the adhesive abilities of the selected strains provides important information about the possible ways to preserve and increase adhesive properties towards epithelium cells.

Some lactic acid bacteria strains in milk media are capable of releasing bioactive peptides. In this study, we evaluated the angiotensin-converting enzyme (ACE)inhibitory activity of 180 lactic acid bacteria and selected several Lactobacillus helveticus, L. delbrueckii subsp. bulgaricus and L. casei strains that demonstrated strong ACE-inhibitory activity. The aim was to carry out a molecular study on the bioactive peptides released by the strains with the best ACE-inhibitory properties and by the strains demonstrating a calcium-binding effect. To the best of our knowledge, this is the first study of bioactive peptides in Bulgarian white cheese. Peptides with the strongest ACE-inhibitory activity were purified and sequenced. The strains were assessed for production of peptides with calcium-binding properties. These peptides were isolated, purified and sequenced. Two strains releasing bioactive peptides with the strongest ACE-inhibitory and calcium-binding activities were selected for development of cheese starters. The strain with the best ACE-inhibitory activity was L. helveticus A1, which releases the peptide Ala-Leu-Pro-Met as a main contributor to the ACE inhibition. The strain with the best calcium-binding activity was L. casei C3 releasing the peptide SpLSpSpSpE (fraction 15–20 of ß-casein) as a main contributor to calcium binding. After pilot production of cheeses with the developed starters, the ACE-inhibitory and calcium-binding effects were confirmed during the cheese ripening. The addition of the two selected adjunct cultures led to increased production of bioactive peptides in the cheese. In this way, it is possible to increase the functional properties of Bulgarian white brined cheese.

Yersiniosis is the third most commonly reported foodborne zoonosis in the European Union. Here, we evaluated the prevalence of pathogenic Yersinia enterocolitica among healthy pigs (as a major reservoir) in a slaughterhouse in Bulgaria. A total of 790 tonsils and feces from 601 pigs were examined. Isolation and pathogenicity characterization was carried out by the ISO 10273:2003 protocol and Polymerase Chain Reaction (PCR), detecting the 16S rRNA gene, attachment and invasion locus (ail), Yersinia heat-stable enterotoxin (ystA), and Yersinia adhesion (yadA) genes. Genetic diversity was assessed by pulsed-field gel electrophoresis (PFGE), and antimicrobial resistance by the standard disk diffusion method. Of all the pigs tested, 6.7% were positive for Y. enterocolitica. All isolates belonged to Y. enterocolitica bioserotype 4/O:3. ail, and ystA genes were detected in all positive strains (n = 43), while the plasmid Yersinia virulence plasmid (pYV) was detected in 41. High homogeneity was observed among the strains, with all strains susceptible to ceftriaxone, amikacin and ciprofloxacin, and resistant to ampicillin. In conclusion, a low prevalence of Y. enterocolitica 4/O:3 was found in healthy pigs slaughtered in Bulgaria, not underestimating possible contamination of pork as a potential risk to consumer health.

The anti-inflammatory and antimicrobial effects of a cream containing extracts of African geranium (Pelargonium sidoides DC.), black elderberry (Sambucus nigra L.) and St. John's wort (Hypericum perforatum L.) in colloidal nanosilver (AgNPs) at a concentration of 30 ppm, denoted as SILVER STOP® cream (SS® cream) was examined In vitro. The research was performed with Staphylococcus aureus (ATCC-6538 and two clinical strains). Suspension tests for determination the time of antimicrobial action of SS® cream were used. SS® cream showed significant antimicrobial activity. In suspension with a density of 104 cells.mL-1S. aureus died after 60 minutes of exposure to SS® cream. In suspension with concentration of 106 cells.mL-1, after 2 h only single cells remained viable. The anti-inflammatory effect was evaluated using human epithelial cell line HT-29, incubated with a type strain of S. aureus. After treatment with SS® cream, results have shown a statistically significant reduction of the levels of the pro-inflammatory cytokinesIL-6, IL-8, IP-10 and IL-1β.