Illuminating the Genomic Landscape of Lactiplantibacillus plantarum PU3-A Novel Probiotic Strain Isolated from Human Breast Milk, Explored through Nanopore Sequencing.

Lactiplantibacillus plantarum stands out as a remarkably diverse species of lactic acid bacteria, occupying a myriad of ecological niches. Particularly noteworthy is its presence in human breast milk, which can serve as a reservoir of probiotic bacteria, contributing significantly to the establishment and constitution of infant gut microbiota. In light of this, our study attempted to conduct an initial investigation encompassing both genomic and phenotypic aspects of the L. plantarum PU3 strain, that holds potential as a probiotic agent. By employing the cutting-edge third-generation Nanopore sequencing technology, L. plantarum PU3 revealed a circular chromosome of 3,180,940 bp and nine plasmids of various lengths. The L. plantarum PU3 genome has a total of 2962 protein-coding and non-coding genes. Our in-depth investigations revealed more than 150 probiotic gene markers that unfold the genetic determinants for acid tolerance, bile resistance, adhesion, and oxidative and osmotic stress. The in vivo analysis showed the strain's proficiency in utilizing various carbohydrates as growth substrates, complementing the in silico analysis of the genes involved in metabolic pathways. Notably, the strain demonstrated a pronounced affinity for D-sorbitol, D-mannitol, and D-Gluconic acid, among other carbohydrate sources. The in vitro experimental verification of acid, osmotic and bile tolerance validated the robustness of the strain in challenging environments. Encouragingly, no virulence factors were detected in the genome of PU3, suggesting its safety profile. In search of beneficial properties, we found potential bacteriocin biosynthesis clusters, suggesting its capability for antimicrobial activity. The characteristics exhibited by L. plantarum PU3 pave the way for promising strain potential, warranting further investigations to unlock its full capacity and contributions to probiotic and therapeutic avenues.

Inhibitory Potential of Different Bilberry (Vaccinium myrtillus L.) Extracts on Human Salivary α-Amylase.

Recently, consumer preferences for bilberries have increased markedly. This fact is probably related to their natural constituents, such as phenolic compounds including anthocyanins and tannins, as well as the vitamins and minerals they contain. Phenolic compounds are known for their numerous beneficial effects on human health. Moreover, bilberry fruits have been shown to inhibit the activity of carbohydrate hydrolyzing enzymes, which can significantly decrease the postprandial increase in blood glucose levels. Thus, the aim of the present study is to investigate the inhibitory effect of Vaccinium myrtillus L. extracts on key enzyme α-amylase, linked to type 2 diabetes. No data have been published on the inhibitory properties of Vaccinium myrtillus L. fruits growing wild in Bulgaria against carbohydrate enzymes. Bilberry extracts were analyzed for total polyphenols, total anthocyanin content, antioxidant activity and their inhibitory properties against α-amylase. The contents of flavonols, anthocyanins and stilbenes were determined by HPLC analysis. The identified flavonols in the analyzed bilberry extracts were mainly represented by quercetin derivatives as rutinoside. The predominant anthocyanins for both aqueous and organic solvents were delphinidin-3-galactoside and malvidin-3-glucoside. The results revealed that bilberry extracts are effective inhibitors of α-amylase, with IC50 values from 20.8 to 194.8 µg GAE/mL. All the samples proved to have antioxidant activity measured by three different in vitro assays (FRAP, CUPRAC and DPPH). The inhibitory properties of V. myrtillus L. extracts may provide a new direction in the development and research of new pharmaceuticals for the suppression of postprandial hyperglycemia in diabetic patients.

The Enzymatic Hydrolysis of Human Milk Oligosaccharides and Prebiotic Sugars from LAB Isolated from Breast Milk.

Breastfeeding is essential in the first months of a newborn's life. Breast milk is a source of crucial macronutrients, prebiotic oligosaccharides, and potential probiotic strains of bacteria. Oligosaccharides from breast milk (HMOs) are a significant part of the composition of breast milk and represent a complex of digestible sugars. This study aims to elucidate the enzymatic hydrolysis of these oligosaccharides and other prebiotics by the bacteria present in breast milk. We used modified methods to isolate oligosaccharides (HMOs) from human milk. Using unique techniques, we isolated and identified different bacteria from breast milk, mainly Lactobacillus fermentum. Using enzymatic analyses, we established the participation of α-fucosidase, α-glucosidase, ß-galactosidase, and ß-glucosidase from breast milk bacteria in the hydrolysis of prebiotic sugars. We also optimized the scheme for isolating oligosaccharides from breast milk by putting the lyophilized product into different food media. We found that the oligosaccharides from breast milk (HMOs) are a potent inducer for the secretion of the studied bacterial enzymes. Also, we found that all the lactobacilli strains we studied in detail could digest mucin-linked glycans. The degradation of these sugars is perhaps a built-in defense mechanism in cases where other sugars are lacking in the environment. We also determined fucosidase activity in some of the isolated strains. We recorded the highest values (2.5 U/mg in L. fermentum ss8) when the medium's oligosaccharides isolated from breast milk were present. Lactobacilli and Bifidobacteria supplied with breast milk are the first colonizers in most cases in the gastrointestinal tract of the newborn. The presence and study of different genes for synthesizing other enzyme systems and transporters of various sugars in this type of bacteria are essential.

Structural Characterization and In Vivo Anti-Inflammatory Activity of Fucoidan from Cystoseira crinita (Desf.) Borry.

The aim of this study was to evaluate the effects of fucoidan isolated from C. crinita on histamine-induced paw inflammation in rats, and on the serum levels of TNF-α, IL-1ß, IL-6, and IL-10 in rats during systemic inflammation response. The levels of TNF-α in a model of acute peritonitis in rats were also investigated. The isolated crude fucoidan was identified as a sulfated xylogalactofucan with high, medium, and low molecular weight fractions and a content of fucose of 39.74%, xylose of 20.75%, and galactose of 15.51%. Fucoidan from C. crinita showed better anti-inflammatory effects in the rat paw edema model, and this effect was present during all stages of the experiment. When compared to controls, a commercial fucoidan from F. vesiculosus, the results also displayed anti-inflammatory activity on the 60th, 90th, and 120th minute of the experiment. A significant decrease in serum levels of IL-1ß in rats treated with both doses of C. crinita fucoidan was observed in comparison to controls, whereas TNF-α concentrations were reduced only in the group treated with fucoidan from C. crinita at the dose of 25 mg/kg bw. In the model of carrageenan-induced peritonitis, we observed a tendency of decrease in the levels of the pro-inflammatory cytokine TNF-α in peritoneal fluid after a single dose of C. crinita fucoidan, but this did not reach the statistical significance margin. Single doses of C. crinita fucoidan did not alter serum levels of the anti-inflammatory cytokine IL-10 in animals with lipopolysaccharide-induced systemic inflammation.

Automatic Landing of Unmanned Aerial Vehicles via Wireless Positioning System with Pseudo-Conical Scanning.

In this work, a wireless UAV unmanned landing system is considered, using the principles of pseudo conical scanning with a phased antenna array (PAA). The basic requirements for the characteristics and parameters of the system as a whole and of its components are defined. Special attention is paid to the primary sensor of the system-PAA with electronic scanning. A variant with a minimum number of four states on the radiation pattern of a low-budget patch PAA was studied. A linear regression of the difference characteristics of the measured radiation beams is proposed, which allows the practical application of the recommended landing algorithm with low computational complexity. Systematic and random positioning errors, both by measurement and by Monte Carlo simulation, were studied. Obtained statistical results prove the algorithm convergence and acceptable accuracy for the system implementation. They are applied if necessary to adjust the Kalman filter parameters. The proposed wireless system can be used for unmanned landing, tracking, and navigating the UAV in flight, or wireless navigation of other mobile objects.

Assessment of the exposure of two pesticides on common carp (Cyprinus carpio Linnaeus, 1758): Are the prolonged biomarker responses adaptive or destructive?

Chlorpyrifos (CPF) and cypermethrin (CYP) are two insecticides that have a proven negative effect on non-target aquatic organisms when they enter the surface waters. However, literature on the comparative effects of these pesticides on important aquaculture fish species, such as common carp (Cyprinus carpio Linnaeus, 1758) is not yet scientifically detailed, especially over the long-term. The idea of conducting a long-term exposure is to find out how the observed biomarkers would change compared to the short-term exposure. In the natural environment, toxicants are not present alone, but in combination. By monitoring the long-term impact of individual substances, the state of aquatic ecosystems exposed to various toxicants could be predicted. Thus, this study aimed to evaluate the toxicity of different concentrations of CYP (0.0002, 0.0003, and 0.0006 µg/L) and CPF (0.03, 0.05, and 0.10 µg/L) in 50-L glass tanks on C. carpio, exposed for 30 days under laboratory conditions. A set of histological and biochemical biomarkers in the gills and liver were applied with the chemical analyses of water and fish organs. Furthermore, the condition and hepatosomatic index were calculated to assess the physiological status of the treated carps. The behavioral responses were also monitored, and the respiration rate was analyzed. The results suggest that CYP had a more prominent effect on the histological structure of fish organs, biochemical responses of anti-oxidant enzymes, behavior, and respiration rate compared to the effect of CPF. In addition, the results also indicate that the liver is more susceptible to chronic and chemically induced cellular stress compared to the gills, with overall destructive changes in the histological biomarkers rather than adaptive. Regardless of the scenario, our results provide novel insights into pesticide exposure and the possible biological impacts on economically important freshwater fish, exposed to lower CYP and CPF concentrations, based on the EU legislation (maximum allowable concentrations, MAC-EQS).

Biomarkers for pollution in caged mussels from three reservoirs in Bulgaria: A pilot study.

The mussel-watch concept was firstly proposed in 1975, which was later adopted by several international monitoring programs worldwide. However, for the very first time, a field experiment with caged mussels was performed in three reservoirs in Bulgaria to follow the harmful effects of sub-chronic pollution (30 days) of metals, trace, and macro-elements, as well as some organic toxicants, such as polybrominated diphenyl ethers and chlorinated paraffins. Therefore, we studied the biometric indices, histochemical lesions in the gills, biochemical changes in the digestive glands (antioxidant defense enzymes, such as catalase, glutathione reductase, and glutathione peroxidase; metabolic enzymes, such as lactate dehydrogenase, alanine aminotransferase, and aspartate aminotransferase, and the neurotransmitter cholinesterase), in addition to the DNA damage in the Chinese pond mussel, Sinanodonta woodiana (Lea, 1834) in Kardzhali, Studen Kladenets and Zhrebchevo reservoirs in Bulgaria. Significant correlation trends between the pollution levels, which we reported before, and the biomarker responses were established in the current paper. Overall, we found that both tested organs were susceptible to pollution-induced oxidative stress. The different alterations in the selected biomarkers in the caged mussels compared to the reference group were linked to the different kinds and levels of water pollution in the reservoirs, and also to the simultaneously conducted bioaccumulation studies.

Which Is More Toxic? Evaluation of the Short-Term Toxic Effects of Chlorpyrifos and Cypermethrin on Selected Biomarkers in Common Carp (Cyprinus carpio, Linnaeus 1758).

The general aim of this study was to investigate the negative short-term effects of different concentrations of chlorpyrifos (CPF) and cypermethrin (CYP), based on the EU legislation (MAC-EQS) in common carp (Cyprinus carpio Linnaeus, 1758) under laboratory conditions and to compare their toxicity. The fish were exposed to the pesticides for 96 h and then different histological and biochemical biomarkers were investigated in the gills and liver, and bioaccumulation analyses were conducted. The chemical studies showed increased pesticide concentrations in the gills as the first site for pollutants compared to the liver at the 96th hour. In addition, the histological analyses showed severe alterations in the gills and liver after exposure to both tested pesticides. In the gills, we found mainly intense proliferative and, to a lesser extent, degenerative changes and alterations in the circulatory system, such as necrosis and vasodilation. In the liver, regressive and progressive lesions, as well as circulatory disturbances and inflammation, were observed. The regressive lesions showed a higher degree of expression compared to the other changes. Furthermore, we found altered enzymatic activities-catalase, glutathione reductase, and glutathione peroxidase-in the liver, compared to the control. Overall, both tested pesticides impacted the studied biomarkers in common carp, even at concentrations lower than those permitted by law. However, the results of the comparative analysis showed a relatively higher toxicity of CYP compared to CPF in the fish. Still, questions persist as to whether the observed changes are adaptive or entirely destructive. To avoid any danger or risk, these pesticides must be applied cautiously, especially near water bodies.

Formation of Free Amino Acids and Bioactive Peptides During the Ripening of Bulgarian White Brined Cheeses.

Bioactive peptides and free amino acids obtained from Bulgarian goat, sheep and cow white brined cheeses, produced with same starter culture, during ripening were evaluated. The concentration of total free amino acids was increasing in all tested cheeses in the first 30 days of ripening. In the next 30 days in sheep cheeses, the concentration increased as recorded for most of the amino acids. Amino acids with highest levels detected throughout the whole ripening period in goat, sheep and cow cheese types were leucine, phenylalanine, arginine, valine and lysine. MALDI-TOF analysis of evaluated cheeses resulted in detection of production of bioactive peptide derivates from milk proteins: 51 peptides in cow, 31 peptides in sheep and 22 peptides in goat cheeses. Peptide αs1-CN (f35-40) was found only in cow cheese. In cow cheese, higher intensity was detected for αs1-CN (f1-9) and ß-CN (f194-203 and f203-219) peptides. In goat cheese was recorded αs1-CN peptides, and there was a tendency to increase the peptides released from ß-CN, with the highest intensity of fragments αs1-CN (f1-9 and f24-30) and ß-CN (f194-209 and f203-219). In sheep cheese, the recorded primarily peptides were αs1-CN and peptides released from ß-CN. Different bioactive peptides, derivate from casein, were detected as follows: 6 peptides were ACE inhibitory peptides, 3 peptides were αS1-casokinins, 1 peptide was caseinophopeptide, 1 peptide was immunopeptide. Twelve bioactive peptides were recorded to be derivates from ß-casein: 1 peptide was ACE peptide, 4 peptides were caseino-phosphopeptides, 1 peptide was immunopeptide, 1 peptide ß-casokinin, 1 antibacterial peptide and 4 multifunctional peptides. Of peptides released by proteolysis of αS2-CN was found 1 bioactive peptide with antimicrobial activity. On our best knowledge, this paper contributes new data about free amino acids and bioactive peptides in the connection between type of milk and period for cheese ripening in the Bulgarian goat, sheep and cow white brined cheeses.

Metabolic Profiling of Xylooligosaccharides by Lactobacilli.

Three lactic acid bacteria (LAB) strains identified as Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus sakei isolated from meat products were tested for their ability to utilize and grow on xylooligosaccharides (XOSs). The extent of carbohydrate utilization by the studied strains was analyzed by HPLC. All three strains showed preferences for the degree of polymerization (DP). The added oligosaccharides induced the LAB to form end-products of typical mixed-acid fermentation. The utilization of XOSs by the microorganisms requires the action of three important enzymes: ß-xylosidase (EC 3.2.1.37) exo-oligoxylanase (EC 3.2.1.156) and α-L-arabinofuranosidase (EC 3.2.1.55). The presence of intracellular ß-D-xylosidase in Lb. brevis, Lb. plantarum, and Lb. sakei suggest that XOSs might be the first imported into the cell by oligosaccharide transporters, followed by their degradation to xylose. The studies on the influence of XOS intake on the lipids of rat liver plasma membranes showed that oligosaccharides display various beneficial effects for the host organism, which are probably specific for each type of prebiotic used. The utilization of different types of oligosaccharides may help to explain the ability of Lactobacillus strains to compete with other bacteria in the ecosystem of the human gastrointestinal tract.

Immunomodulatory and Anti-Inflammatory Effects of Fucoidan: A Review.

Inflammation is the initial response of the immune system to potentially harmful stimuli (e.g., injury, stress, and infections). The process involves activation of macrophages and neutrophils, which produce mediators, such as nitric oxide (NO), prostaglandin E2 (PGE2), pro-inflammatory and anti-inflammatory cytokines. The pro-inflammatory cytokines interleukin-1ß (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) are considered as biomarkers of inflammation. Even though it occurs as a physiological defense mechanism, its involvement in the pathogenesis of various diseases is reported. Rheumatoid arthritis, inflammatory bowel disease, Alzheimer's disease, and cardiovascular diseases are only a part of the diseases, in which pathogenesis the chronic inflammation is involved. Fucoidans are complex polysaccharides from brown seaweeds and some marine invertebrates, composed mainly of L-fucose and sulfate ester groups and minor amounts of neutral monosaccharides and uronic acids. Algae-derived fucoidans are studied intensively during the last years regarding their multiple biological activities and possible therapeutic potential. However, the source, species, molecular weight, composition, and structure of the polysaccharides, as well as the route of administration of fucoidans, could be crucial for their effects. Fucoidan is reported to act on different stages of the inflammatory process: (i) blocking of lymphocyte adhesion and invasion, (ii) inhibition of multiple enzymes, and (iii) induction of apoptosis. In this review, we focused on the immunemodulating and anti-inflammatory effects of fucoidans derived from macroalgae and the models used for their evaluation. Additional insights on the molecular structure of the compound are included.

Physiological and molecular insights of bacteriocin production by Enterococcus hirae ST57ACC from Brazilian artisanal cheese.

The bacteriocinogenic Enterococcus hirae ST57ACC recently isolated from a Brazilian artisanal cheese was subjected here to additional analyses in order to evaluate its bacteriocin production and the potential influence of ABC transporter system in its expression. Besides these physiological and molecular aspects, the bacteriocin was evaluated for its cytotoxicity against HT-29. Differences in the inoculum size had no impact on the growth of E. hirae ST57ACC; however, the bacteriocin was only produced after 9 h of growth when the strain was inoculated at 5% or 10% (v/v), with similar levels of bacteriocin production obtained by both conventional growth and batch fermentation. Furthermore, potential expression of ABC transporters corresponding to the bacteriocin transport and sugar metabolism was identified. In terms of adverse effects, when a semi-purified fraction of the bacteriocin and the cell-free supernatant were tested against HT-29, total cell viability was similar to observed on untreated cells, indicating the absence of cytotoxic effect. Based on the obtained results, E. hirae ST57ACC can produce its bacteriocin at industrial level by using bioreactors, its bacteriocin expression is potentially influenced by the ABC transporter system, and no cytotoxic effects were observed on HT-29 cells, indicating its potential use as a bio-preservative.

Opioid-Induced Hyperalgesia in the Nonsurgical Setting: A Systematic Review.

BACKGROUND: Opioid-induced hyperalgesia (OIH) is a phenomenon that causes an increased pain sensitization and perception of pain to noxious stimuli secondary to opioid exposure. While this clinical effect has been described in the surgical setting, it is unclear if OIH occurs in the nonsurgical setting. STUDY QUESTION: To review the available literature which evaluated OIH in nonsurgical settings. DATA SOURCES: A comprehensive literature search was performed using PubMed (January 1946-July 2017) using a variety of keywords for OIH. This review included randomized controlled trials with objectives to identify OIH in the nonsurgical setting. The clinical outcomes of interest were identification of OIH, adverse events, and impact of OIH on opioid consumption. RESULTS: The search identified 8 studies that fulfilled the criteria. Six studies enrolled healthy male volunteers, 1 study used chronic low-back patients, and another used heroin-dependent treatment-seeking adults. Studies used various opioids and dosages, including remifentanil, alfentanil, fentanyl, morphine, methadone, and buprenorphine. Three primary experimental pain induction models were used to evaluate for OIH. Measured outcomes included hyperalgesia area, pain threshold, and pain tolerance. All 5 studies that used the electrical stimulation model identified OIH as a significant outcome. However, only 2 of 5 studies using the cold pressor model and 1 of 3 studies using the heat pressor model identified OIH. None of the trials explored clinical outcomes, such as effects on opioid consumption. CONCLUSIONS: Most included studies identified OIH as a significant outcome within the nonsurgical setting. However, due to conflicting conclusions and various limitations, the clinical impact of OIH could not be assessed. Clinicians should monitor for effects of OIH in the nonoperative setting because there is insufficient evidence from the available literature to conclude that OIH is consistently observed in this setting.

Study of prebiotic potential and antioxidant activity in Plantago spp. leaves after enzymatic hydrolysis with hemicellulase and xylanase.

The leaves of Plantago species have been known as a rich source of polysaccharides, polyphenols, and iridoids. In the recent years, some authors have examined the relative influence of carbohydrate compositions and polyphenols on their antioxidant and prebiotic activity. In this study enzymatic hydrolysis of Plantago major, Plantago lanceolata, and Plantago media leaves with hemicellulase and xylanase was carried out. The chemical composition of the enzyme-derived hydrolysis products was determined. Their antioxidant activity was evaluated using 2,2-diphenyl-2-picryl-hydrazyl-hydrate, ferric reducing antioxidant power, and cupric reducing antioxidant capacity assays. The utilization of the carbohydrate complex from the obtained hydrolysates to stimulate the production of certain glycohydrolases from Lactobacillus plantarum probiotic strains was investigated. The HPLC analysis of the hydrolysis products showed the presence of galactose and arabinose in 1:3 and 1:4 ratios. The highest concentration of total sugars was determined in P. major hydrolysates with hemicellulase (386.71 mg/g dry weight). The antioxidant activity according to the three methods was the highest in P. media hydrolysates, which correlated with their total phenolic concentration. L. plantarum S27 showed the highest values of α-galactosidase activity at the 24-h fermentation process when it was cultivated in a broth with the presence of P. major hydrolysates with hemicellulase and xylanase (0.129 U/mg and 0.133 U/mg, respectively).

Synthesis of glucooligosaccharides with prebiotic potential by glucansucrase URE 13-300 acceptor reactions with maltose, raffinose and lactose.

In the present work, we report an efficient synthesis of glucooligosaccharides (GOSs) with prebiotic potential by novel glucansucrase URE 13-300 from Leuconostoc mesenteroides URE 13 strain. The highest total yield of GOSs with degree of polymerization (DP) from 3 to 6 was obtained with maltose as an acceptor and maltose/sucrose (M/S) ratio 1-136 g/L. An efficient modulation of GOSs composition is achieved by varying the M/S ratio. At M/S = 1, 2, 4 and 7 the content of DP3 products gradually increase from 54.50 to 91.70%. When the M/S ratio was decreased the synthesis of DP>3 GOSs is predominant and reaches 75.60% (M/S = 0.25). In addition, the maltose derived GOSs with DP>3, as well as raffinose and lactose glucosylation products have a branched structure which is prerequisite for increased prebiotic potential. The synthesized GOSs were efficiently metabolized by probiotic strains of Lb. plantarum S26, Lb. brevis S27 and Lb. sakei S16, and the calculated values of specific growth rate (µ) were nearly identical to this on glucose media, when maltose derived GOSs were used as a carbohydrate source. Strain specific features were observed in the utilization of the synthesized GOSs, as well as in the production of lactic acid and acetic acid.

Enzymatic Hydrolysis of Water Extractable Polysaccharides from Leaves of Plantago major L.

BACKGROUND: Plantago major L. leaves have been used for centuries by the traditional medicine in the treatment of infectious disorders of the respiratory, urinary and digestive tracts. Researchers have reported that hot water extracts of Plantago major possess a broad-spectrum of anticancer, antioxidant and antiviral activities, as well as activities which modulate cell-mediated immunity. Their beneficial properties may be due to the significant content of polysaccharides. The polysaccharides that have been isolated from the leaves of Plantago major L. have different structures - pectic substances, galactans, arabinogalactans, glucomannans. AIM: The aim of this paper was to study the correlation between the structure of the water extractable polysaccharides isolated from Plantago major L. leaves and their enzymatic hydrolysis with different carbohydrate hydrolases. MATERIALS AND METHODS: The hydrolysis reactions were performed with the enzymes hemicellulase and mannanase. Spectrophotometric total reducing sugars assay was used to examine the hydrolysis yield. The monosaccharide and oligosaccharide compositions were determined using HPLC analysis. RESULTS: The highest hydrolysis yield of the water extractable polysaccharides from Plantago major leaves was obtained by treatment with hemicellulase. The hydrolysis yield increased with the augmentation of the ratio of enzyme to polysaccharide. Galactose was the prevalent monosaccharide identified in the composition of the isolated polysaccharides. Oligosaccharides with different degree of polymerization were also detected. CONCLUSION: The enzymatic hydrolysis of water extractable polysaccharides from Plantago major leaves allows us to obtain different types of oligosaccharides with beneficial effects on both human health and industry.

Effect of Extracts of Bilberries (Vaccinium myrtillus L.) on Amyloglucosidase and α-Glucosidase Activity.

BACKGROUND: Vaccinium myrtillus L. is a species belonging to the genus Vaccinium of the family Ericaceae. Bilberries have drawn attention due to the multiple benefits for the human health, including antioxidant, anti-inflammatory, anticancer, anti-neurodegenerative, and cardioprotective effects. Recently, bilberries were shown to inhibit the activity of carbohydrate-hydrolysing enzymes that can help reduce the intensity of the metabolic syndrome and prevent type 2 diabetes. AIM: In this study, we investigated the α-glucosidase and amyloglucosidase inhibitory activities of polyphenol-rich extracts from fruit of Vaccinium myrtillus L. from different regions in Bulgaria. MATERIALS AND METHODS: The total phenolic content was determined spectrophotometrically using the Folin-Ciocalteu method. With HPLC analysis, phenolic acid composition of extracts was assessed. Enzymatic inhibitory activities were determined according to the methodology by Borooah et al. (1961), and Dewi et al. (2007). Amyloglucosidase assay and α-glucosidase assay were used to measure the inhibition potential of bilberries' extracts. RESULTS: Phenolic compound content ranged from 1299.60 mg to 510.88 mg GAE/100 g for organic extracts and from 453.63 mg to 290.83 mg GAE/100 g for aqueous extracts. Based on qualitative HPLC analyses, gallic acid and chlorogenic acid were found to be among the major phenolic acids present in bilberries. Methanol and aqueous extracts there were found to be effective inhibitors of α-glucosidase with an IC50 value of 20 µg GAE/ml and 55 µg GAE/ml, respectively. CONCLUSION: The inhibitory activity of bilberries' extracts towards α-glucosidase offers the patients with type 2 diabetes the opportunity to manage their own glycaemic levels with a diet.

Technology and safety assessment for lactic acid bacteria isolated from traditional Bulgarian fermented meat product "lukanka"

Abstract The present work discusses the technological and new selection criteria that should be included for selecting lactic acid bacteria for production of fermented meat. Lactic acid bacteria isolated from Bulgarian traditional fermented "lulanka" salami was studied regarding some positive technological parameters (growth at different temperature, pH, and proteolytic activity). The presence of genes related to the virulence factors, production of biogenic amines, and vancomycin resistance were presented in low frequency in the studied lactic acid bacteria. On the other hand, production of antimicrobial peptides and high spread of bacteriocin genes were broadly presented. Very strong activity against L. monocytogenes was detected in some of the studied lactic acid bacteria. In addition, the studied strains did not present any antimicrobial activity against tested closely related bacteria such as Lactobacillus spp., Lactococcus spp., Enterococcus spp. or Pediococcus spp. To our knowledge this is the first study on the safety and antimicrobial properties of lactic acid bacteria isolated from Bulgarian lukanka obtained by spontaneous fermentation.

Technology and safety assessment for lactic acid bacteria isolated from traditional Bulgarian fermented meat product "lukanka".

The present work discusses the technological and new selection criteria that should be included for selecting lactic acid bacteria for production of fermented meat. Lactic acid bacteria isolated from Bulgarian traditional fermented "lulanka" salami was studied regarding some positive technological parameters (growth at different temperature, pH, and proteolytic activity). The presence of genes related to the virulence factors, production of biogenic amines, and vancomycin resistance were presented in low frequency in the studied lactic acid bacteria. On the other hand, production of antimicrobial peptides and high spread of bacteriocin genes were broadly presented. Very strong activity against L. monocytogenes was detected in some of the studied lactic acid bacteria. In addition, the studied strains did not present any antimicrobial activity against tested closely related bacteria such as Lactobacillus spp., Lactococcus spp., Enterococcus spp. or Pediococcus spp. To our knowledge this is the first study on the safety and antimicrobial properties of lactic acid bacteria isolated from Bulgarian lukanka obtained by spontaneous fermentation.

Glucansucrases produced by fructophilic lactic acid bacteria Lactobacillus kunkeei H3 and H25 isolated from honeybees.

Twenty fructophilic isolates from the stomachs of honeybee Apis mellifera ligustica from the region of Plovdiv, Bulgaria were obtained. Fructophilic isolates H3 and H25 showed formation of mucous colonies during cultivation on medium with sucrose, suggesting exopolysaccharide synthesis. The sequencing analysis of 16S rRNA identified isolates H3 and H25 as fructophilic lactic acid bacteria Lactobacillus kunkeei. The in situ analysis and periodic acid-Schiff's staining, showed that Lb. kunkeei H3 and H25 produce extracellular glucansucrases with molecular weight of about 300 kDa. In the cell-associated fractions, additional glucansucrase is detected with molecular weight of about 180 kDa. The content of α-(1 → 6) linkages in the glucans synthesized with extracellular glucansucrases from H3 and H25 after dextranase hydrolysis was significantly lower than this one of the classical dextran - about 35 and 62%, respectively. These results suggest a more branched structure of the studied polymers.