Search results for: metabiotics

Authors: A. M. Isakhanyan, F. N. Tkhruni, N. N. Yakimovich, Z. I. Kuvaeva, T. V. Khachatryan

Abstract:

Introduction: At present, the simbiotics based on different genera and species of lactic acid bacteria (LAB) and yeasts are used. One of the basic properties of probiotics is presence of antimicrobial activity and therefore selection of LAB and yeast strains for their co-cultivation with the aim of increasing of the activity is topical. Since probiotic yeast and bacteria have different mechanisms of action, natural synergies between species, higher viability and increasing of antimicrobial activity might be expected from mixing both types of probiotics. Endemic strains of LAB Enterococcus faecium БТK-64, Lactobaccilus plantarum БТK-66, Pediococcus pentosus БТK-28, Lactobacillus rhamnosus БТK-109 and Kluyveromyces lactis БТX-412, Saccharomycopsis sp. БТX- 151 strains of yeast, with probiotic properties and hight antimicrobial activity, were selected. Strains are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB and yeast strains were isolated from different dairy products from rural households of Armenia. The genotyping by 16S rRNA sequencing for LAB and 26S RNA sequencing for yeast were used. Combined cultivation of LAB and yeast strains was carried out in the nutrient media on the basis of milk whey, in anaerobic conditions (without shaker, in a thermostat at 37oC, 48 hours). The complex preparations were obtained by purification of cell free culture broth (CFC) broth by the combination of ion-exchange chromatography and gel filtration methods. The spot-on-lawn method was applied for determination of antimicrobial activity and expressed in arbitrary units (AU/ml). Results. The obtained data showed that at the combined growth of bacteria and yeasts, the cultivation conditions (medium composition, time of growth, genera of LAB and yeasts) affected the display of antimicrobial activity. Purification of CFC broth allowed obtaining partially purified antimicrobial complex preparation which contains metabiotics from both bacteria and yeast. The complex preparation inhibited the growth of pathogenic and conditionally pathogenic bacteria, isolated from various internal organs from diseased animals and poultry with greater efficiency than the preparations derived individually alone from yeast and LAB strains. Discussion. Thus, our data shown perspectives of creation of a new class of antimicrobial preparations on the basis of combined cultivation of endemic strains of LAB and yeast. Obtained results suggest the prospect of use of the partially purified complex preparations instead antibiotics in the agriculture and for food safety. Acknowledgments: This work was supported by the RA MES State Committee of Science and Belarus National Foundation for Basic Research in the frames of the joint Armenian - Belarusian joint research project 13РБ-064.

Keywords: co-cultivation, antimicrobial activity, biosafety, metabiotics, lactic acid bacteria, yeast

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Authors: Kristina Karapetyan, Flora Tkhruni, Tsovinar Balabekyan, Arevik Israyelyan, Tatyana Khachatryan

Abstract:

The ability of the lactic acid bacteria (LAB) to prevent and cure a variety of diseases, their protective role against infections and colonization of pathogenic microorganisms in the digestive tract, has lead to the coining of the term probiotics or pro-life. LAB inhibiting the growth of pathogenic and food spoilage microorganisms, maintaining the nutritive quality and improving the shelf life of foods. They have also been used as flavor and texture producers. Enterococcus strains have been used for treatment of diseases such as diarrhea or antibiotic associated diarrhea, inflammatory pathologies that affect colon such as irritable bowel syndrome, or immune regulation, diarrhea caused by antibiotic treatments. The obtaining and investigation of biological properties of proteinoceous antibiotics, on the basis of probiotic LAB shown, that bacteriocins, metabiotics, and peptides of LAB represent bactericides have a broad range of activity and are excellent candidates for development of new prophylactic and therapeutic substances to complement or replace conventional antibiotics. The genotyping by 16S rRNA sequencing for LAB were used. Cell free culture broth (CFC) broth was purified by the Gel filtration method on the Sephadex Superfine G 25 resin. Antimicrobial activity was determined by spot-on-lawn method and expressed in arbitrary units (AU/ml). The diversity of multidrug-resistance (MDR) of pathogenic strains to antibiotics, most widely used for treatment of human diseases in the Republics of Armenia and Nagorno Karabakh were examined. It was shown, that difference of resistance of pathogens to antibiotics depends on their isolation sources. The influences of partially purified antimicrobial preparations (AMP), obtained from the different strains of Enterococcus genus on the growth of MDR pathogenic bacteria were investigated. It was shown, that bacteriocin containing partially purified preparations, obtained from different strains of Enterococcus faecium and durans species, possess bactericidal or bacteriostatic activity against antibiotic resistant intestinal, spoilage and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus, E. coli, and Salmonella. Endemic strains of LAB, isolated from Matsoni made from donkey, buffalo and goat milk, shown broad spectrum of activity against food spoiling microorganisms, moulds and fungi, such as Salmonella sp., Esherichia coli, Aspergillus and Penicillium species. Highest activity against MDR pathogens shown bacteria, isolated from goat milk products. High stability of the investigated strains of the genus Enerococcus, isolated from samples of matsun from different regions of Nagorno-Karabakh (NKR) to the antibiotics was shown. The obtained data show high stability of the investigated different strains of the genus Enerococcus. The high genetic diversity in Enterococcus group suggests adaptations for specific mutations in different environments. Thus, endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms isolated from different sources and belong to different taxonomic group. Prospect of the use of certain antimicrobial preparations against pathogenic strains is obvious. These AMP can be applied for long term use against different etiology antibiotic resistant pathogens for prevention or treatment of infectional diseases as an alternative to antibiotics.

Keywords: antimicrobial biopreparation, endemic lactic acid bacteria, intra-species diversity, multidrug resistance of pathogens

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