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Scientific Report 2015 Bulgarian Academy of Sciences The Stephan Angeloff Institute of Microbiology Member of the Institut Pasteur International Association SCIENTIFIC REPORT 2015 The Stephan Angeloff Institute of Microbiology Bulgarian Academy of Sciences 26, Acad. G. Bonchev Str. 1113 Sofia BULGARIA Tel/Fax: +359-2 70 01 09 E-mail: [email protected] www.microbio.bas.bg 2 Prof. Hristo Najdenski, DVM, DSc, Corresponding Member of BAS Director Assoc. Prof. Ivan Simeonov, PhD Assoc Prof. Lyubka Doumanova, PhD Deputy-Director Scientific Secretary Prof. Nina Ivanovska, DSc Chairman of General Assembly Assoc Prof. Zlatka Alexieva, PhD Chairman of Scientific Council 3 CONTENT Scientific Reports on Projects and Publications 5 Departament of General Microbiology 6 Departament of Infectious Microbiology 8 Departament of Applied Microbiology 13 Departament of Virology 22 Departament of Immunology 29 Section of Mycology 33 List of Publications not included in the Scientific Projects 37 Education Activity 43 Scientific Events 47 Staff of the Stephan Angeloff Institute of Microbiology 50 Scientific Board 53 International Scientific Board 54 National Scientific Seminars 55 Annex 1 - Photos of Important Events 56 4 SCIENTIFIC PROJECT REPORTS PUBLICATIONS ACTIVITIES 5 DEPARTMENT OF GENERAL MICROBIOLOGY SECTION OF MICROBIAL GENETICS Current projects MODERN MOLECULAR APPROACHES FOR FUNCTIONAL CHARACTERIAZATION OF THE PROBIOTIC PROPERTIES AND TECHNOLOGICAL CHARACTERISTICS STUDY OF THE AMYLOLYTIC LACTIC ACID BACTERIA FROM BULGARIAN FERMENTATIVE PRODUCTS Project leader: V. Gotcheva, PhD, University of Food Technologies, Plovdiv, Bulgaria Project collaborators: The Stephan Angeloff Institute of Microbiology, BAS, Sofia; Institute of Chemical Engineering, BAS, Sofia Research staff: A. Angelov, PhD, G. Blagoeva; P. Stefanova, P. Petrova, PhD, P. Velikova, A. Stoyanov, PhD; K. Petrov, PhD; F. Tsvetanova; L. Popova; E. Vassileva, PhD Amylolytic lactic acid bacteria glgB, agl, malL, treC, and dexC were (ALAB) are diverse group of examined. Although all strains possessed microorganisms that are capable to degrade extracellular and cell-bound amylase activity starchy materials and to convert them and produced lactic acid from starch, high directly into sugars with lower molecular genus and species specificity in the gene weight, lactic acid, and volatiles. They have expression was observed. ALAB strains of numerous applications in food industry due genus Lactobacillus (except L. sakei) and P. to specific ability to improve the acidilactici own and express all the tested organoleptic properties and to increase the genes, while E. faecium and E. durans nutritional value of starchy sources foodstuff strains expressed predominantly the gene, in course of fermentation processing, and encoding amylase. The co-transcription of are also employed in perspective glgP and glgB genes indicates that glycogen biotechnologies for lactic acid production synthesis and starch degradation occur in using renewable and abundant starch parallel, which is another example for dual biomass as a feedstock. During the current metabolic role of biochemical paths. project, 100 strains of ALAB were isolated Other purpose of the current project from amylaceous fermented foods, cereals was the elucidation of probiotic-prebiotics and beverages. Twenty five new ALAB interaction. Two β-fructosidases of L. strains, belonging to 11 distinct species of paracasei strains (involved in inulin and four genera were isolated and analyzed. oligo-fructose metabolism) were Among them, the first amylolytic overproduced, purified and compared. The Lactobacillus sakei, Enterococcus faecium analysis of their biochemical properties and E. durans were reported. The presence showed similar pH, temperature optimum and expression of the genes amy1, glgP, and stability. Substrate specificity revealed 6 that InuB41 had the highest affinity to short- achieved. These yield and conversion rates chain inulin, while InuLC1 possessed the are the highest obtained by SSF for LA highest affinity to levan. Inulin degradation production from inulin. profiles of the enzymes showed the presence of fructose and sucrose only, thus proving Grant DFNI B02/27-2014, National Science the enzymes affiliation to exo-inulinases Fund (EC 3.2.1.80). Highly effective bio-process for lactic acid (LA) production by References simultaneous saccharification and Petrova, P., Velikova, P., Popova, L., fermentation (SSF) of chicory flour was Petrov, K. (2015). Direct conversion of developed. In batch fermentation with chicory flour into L(+)-lactic acid by optimized medium content and fermentation the highly effective inulinase producer conditions, a complete conversion of 136 Lactobacillus paracasei DSM 23505. g/L chicory flour into 123.7 g/L LA was Bioresour. Technol. 186, 329–333. ISOLATION, IDENTIFICATION AND CHARACTERIZATION OF LACTOBACILLUS STRAINS FROM UNIQUE HOMEMADE BULGARIAN FERMENTED MILK PRODUCTS Project leader: P. Petrova, PhD Project collaborators: Institute of Chemical Engineering, BAS, Sofia Research staff: P. Velikova; A. Stoyanov, PhD; R. Eneva, PhD; E. Kroumova, PhD; J. Miteva- Staleva, MS; M. Zaharieva, PhD; K. Petrov, PhD; L. Popova; H. Najdenski, DSc The aim of the project is to acid bacteria and their genetic and investigate the microbial diversity of biochemical characterization are in progress. artisanal fermented milk products from isolated and remote territory. The selection Grant from the Bright Dairy & Foods Co. of the available strains, belonging to Lactic Ltd, China. 7 DEPARTMENT OF INFECTIOUS MICROBIOLOGY LABORATORY OF ZOONOZES AND BACTERIAL VIRULENCE Current project: DETECTION OF YERSINIA ENTEROCOLITICA IN PIG FAECES BY USING LOOP MEDIATED DNA AMPLIFICATION Project coordinator: H. Najdenski, DSc Collaborators: Unit “Food Safety”, Institute for Agriculture and Fisheries Research (ILVO), Melle, Belgium Research staff: M. Zaharieva, PhD; M. Gatzovska, MS; E. van Collie, PhD; M. Heyndricks, PhD; L. Herman, PhD Yersiniosis is the third most from tissue samples were compared. Finally, commonly reported zoonosis in the two kits for DNA - QIAamp DNA Stool European Union. Consumption of Mini Kit (Qiagen) and DNA Stool isolation contaminated pork meat is important kit (EURx) were used for the investigations. transmission route. Current detection Artificial contamination of farm samples problems are related to the difficult was performed in order to optimize the processing of meat samples and time reaction conditions and to evaluate the consuming bacteriological methods. sensitivity of LAMP methodology. The Detection methods based on conventional products of the LAMP reaction were PCR end-point analysis are faster but low visualized with hydroxynaphthol blue and sensitive. Aim of our study was to develop DNA electrophoresis in agarose gel. The fast, simple, low-cost but specific and sensitivity of LAMP was compared to that sensitive procedure for identification of of TaqMan-qPCR and classical CFU Yersinia enterocolitica in pig faeces by enumeration on agar plates after artificial using loop mediated DNA amplification contamination of fresh pig faeces. (LAMP). The primer set targeting phoP showed Two primers’ sets, complementary to selectivity only for Y. enterocolitica strains. the genes gyrB and phoP were tested for The sensitivity of LAMP was equal to that selectivity on 6 pathogenic Y. enterocolitica of TaqMan-qPCR – 1÷10 DNA copies per strains (serotypes О:3, О:5, О:8, О:9), 10 reaction after pure DNA input and 1.6x104 Yersinia spp. strains and 16 other Gram (-) cfu/mL after artificial contamination. The and Gram (+) bacterial species. The detection limit depends strongly on the DNA sensitivity of LAMP was estimated by isolation kit used, whereby the DNA Stool amplifying of serial dilution of pure isolation kit (EURx) was determined as most bacterial DNA isolated according to the sensitive. The hydroxynaphthol blue protocol of Flamm (1984) from the well visualisation corresponded to the gel characterized Y. enterocolitica strain IP684 electrophoresis result. In conclusion, the (O:8). Five different kits for DNA isolation LAMP method is reliable, fast, cheap, 8 selective and sensitive, whereby the pathogens based on colour visualization of sensitivity is directly related to the DNA the reaction product after appropriate isolation method used. It could be applied optimizations of the reaction conditions. for direct identification of food-borne Belgian-Bulgarian Bilateral project ANTIBACTERIAL, RADICAL-SCAVENGING AND ACETYLCHOLINESTERASE INHIBITORI ACTIVITY OF THE PLANT SPECIES SOLANUM CARENSE И GEIGERIA ALATA USED IN SUDANESE FOLK MEDICINE Project supervisor: G. Momekov, PhD, Department of Pharmacognosy, Faculty of Pharmacy, University of Sofia Project leader: H. Najdenski, DSc Collaborators: The Stephan Angeloff Institute of Microbiology, BAS; Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Sofia Research staff: M. Zaharieva, PhD; R. Gevrenova, PhD; D. Zheleva, PhD; V. Balabanova- Buzushka, PhD Solanum carense and Geigeria alata roots and leaves were obtained by HR- are traditional plants used in Sudanese folk ESI/MS. Major compounds of G. alata roots medicine for treatment of diabetes, cough, were isolated using low-bar liquid epilepsy, intestinal complaints or infections. chromatography. Quantitative analysis of In the frame of this project bithh plant phenolic acids was performed by validated species were chemically analyized and HPLC-UV with limits of detection ranged evaluated for their antioxidant and
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