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

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

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

SCIENTIFIC PROJECT REPORTS PUBLICATIONS ACTIVITIES

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.

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 from 0.04 µg/ml to 0.57 µg/ml. For the first antimicrobial potential. time protocatechuic, caffeic, p- A newly discovered Nε-feruloyl lysine coumaroylquinic, caffeoylsinapoylquinic, (1) together with 16 known caffeoylferuloylquinic, three feruloylquinic, hydroxycinnamic acid amides (HCAAs) six caffeoylquinic acids, and caffeic acid were identified in the roots of Sudanese hexoside were analyzed by HR-MS in G. plant Solanum schimperianum Hochst (syn. alata roots and leaves. HPLC-UV Solanum carense Dunal, Solanaceae). The determination revealed the highest content structure of 1 was established on the basis of of 3,5-dicaffeoylquinic acid (25.96±2.08 1D (1H, 13C NMR) and 2D (COSY, HSQC) mg/g dry weight) in roots, while 4,5- NMR analysis, together with UV–Vis, HR- dicaffeoylquinic acid was the main ESI-MS and ESI-MS2 fragmentation compound in leaves (8.99 ± 0.56 mg/g dw). analyses. The 16 HCAAs were analyzed In total, the amount of caffeoylquinic acids using liquid chromatography coupled to reached up to 6.22% in G. alata roots. mass spectrometry in parallel-reaction Free radical scavenging DPPH, ABTS monitoring (PRM) mode (Orbitrap and ferric reducing power (FRAP) methods ThermoScientific). Eight HCAAs including were used for antioxidant activity evaluation agmatine and cadaverine amides, as well as of Nε-feruloyl lysine, a hydromethanolic Nε-feruloyl lysine, and sinapoyl putrescine extract of S. shimperianum roots and G. were reported in genus Solanum for the first alata roots and leaves extracts. The time. antimicrobial activity was estimated on a Phenolic acids profiles of the panel of pathogenic bacteria and fungi by methanol-aqueous extracts from G. alata the broth microdilution method (BMD).

Nε-feruloyl lysine demonstrated scavenging activity and reducing power stronger DPPH (IC50 85.83 μg/mL), ABTS compared to the crude G. alata extracts and (IC50 255.54 μg/mL) and FRAP (0.44±0.02 pure chlorogenic acid (reference control). mM TE/mg dw) as compared with the total The 3,4,5-tricaffeoylquinic acid revealed S. schimperianum extract. The total S. higher antibacterial potential against the schimperianum extract showed penicillin sensitive and resistant S. aureus antimicrobial activity towards S. pyogenes, strains, as well as towards MRSA as Listeria monocytogenes and Candida compared to the crude G. alata extracts and albicans (MIC 1.25 mg/ml). Nε-feruloyl 3,5-dicaffeoylquinic acid. lysine exhibited significantly higher In conclusion both species represent antimicrobial activity against both new powerful source of biological active Streptococcus aureus strains NBIMCC 3359 substances with strong antioxidant potential (MIC/MBC 112.5 μg/ml ) and penicillin of certain compounds and selective resistant ATCC 6538 P (MIC 112.5 μg/ml) antimicrobial activity against Gram-positive than the total S. schimperianum extract bacterial pathogens. (MIC 1.25 and 2.5 mg/ml, respectively). The 3,5-dicaffeoylquinic acid isolated “Grant-2015” of the Council for Medical from G. alata demonstrated stronger radical Science, Medical University of Sofia

PREVALENCE OF HUMAN PATHOGENIC YERSINIA ENTEROCOLITICA IN TONSILS OF SLAUGHTER-AGED PIGS

Project supervisor: H. Najdenski, DSc Project collaborators: Unit “Food Safety”, Institute for Agriculture and Fisheries Research (ILVO), Melle, Belgium Research staff: M. Gatzovska, Ms, M. Zaharieva, PhD, I. Tsvetkova, Ms, L. Dimitrova, PhD student The aim of the study was to collect enterocolitica and Y. pseudotuberculosis preliminary data concerning the prevalence were applied (according Lambertz, 2008). of human enteropathogenic Yersinia on the This study was completed by bioserotyping tonsils and feces of slaughtered pigs in of isolated Yersinia strains in Yersinia Bulgaria and to determinate the regional Reference Center at Pasteur Institut (Paris, distribution of these pathogens. Tonsil France). samples of 104 fattening pigs, as well 45 Of the tested pigs 20,19% were found feces samples were collected at a positive for Yersinia enterocolitica of which slaughterhouse near Kostinbrod, Bulgaria. 7.69% were proved as pathogenic Y. Modified Horizontal method for the enterocolitica biotype 4, serotype O:3, detection of presumptive pathogenic lysotype VIII. The percentage of positive Yersinia enterocolitica was used. Two days pigs of human Y. enterocolitica pathogenic enrichment in Peptone Sorbitol Bile Salt strains between Stara Zagora’s and Sofia’s broth at 26oC was applied, followed by regions show no difference. No bacteria streaking on Selective Cefsulodin-Irgasan- from the species Y. pseudotuberculosis were Novobiocin (CIN) agar after alkali detected. In conclusion, the study confirms treatment. Typical colonies were confirmed the prevalence of human enteropathogenic by biochemical methods and DNA isolation Y. enterocolitica as a hazard and potential of crude cell lysates was carried out. For the risk for cross-contamination and likewise for PCR identification specific primers for food safety and human health. At slaughter 16SrDNA and chromosomal ail gene for Y. level classical tonsils detection of human

10 pathogenic Y. enterocolitica should be Belgian-Bulgarian Bilateral project completed by more sensitive molecular methods.

PARATUBERCULOSIS IN ANIMALS AND HUMANS –ACTUAL HEALTH AND ECONOMIC PROBLEM

Project supervisor: T. Savova, PhD, National Diagnostic and Research Veterinary Institute “Prof. G. Pavlov” Project leader: M. Bonovska, PhD Project collaborators: The Stephan Angeloff Institute of Microbiology, BAS; National Diagnostic and Research Veterinary Institute “Prof. G. Pavlov”, Sofia, Bulgaria Research staff: H. Najdenski, DSc; V. Valcheva, PhD Mycobacterium avium subspecies (bold, strongly folded and soft intestinal paratuberculosis is the etiological agent of wall, with hyperemia). Histological analysis paratuberculosis in animals (Johne’s revealed the proliferative inflammation of disease) and in humans (Crohn's disease). the mucosa, granulomas without For more than three decades, official caseousation, macrophages with frothy veterinary medical statistics in Bulgaria does cytoplasm and large number of epithelioid not include data on the prevalence of cells in the absence of giant cells. The paratuberculosis in animals due to its rare microscopical, bacteriological and molecular notification. There are no investigations and tests show the presence of Mycobacterium National program for control and monitoring avium subspecies paratuberculosis. These of this disease in both domestic and wild results indicate the need of systematic study animals. Nevertheless, several cases of on the prevalence of this disease in domestic paratuberculosis in a dairy farm were and wild animals in Bulgaria. That is why registered in 2014 in Northeastern Bulgaria. the application of combination of rapid Diseased cows demonstrated diarrheal methods will allow developing of syndrome, birth of non-viable calves or methodology for control of paratuberculosis presence of postnatal complications, severe in Bulgaria, to conduct epidemiological dehydration and death. Furthermore, in analysis and to assess the risk to animals and recent years, the importation of cattle from humans. other countries has increased significantly and therefore early and reliable diagnosis Joint research project between NDRVI and and the epidemiological status concerning SAIM, BAS this zoonotic disease in our country is References: imperative. 50 samples from wild animals Bonovska, M., Savova, T., Valcheva, V., (shot and dead deer and mouflons, grown Najdenski, H. (2015). Paratuberculosis freely in a hunting area in the Sofia region, Bulgaria) were obtained in the period 2009- in animals and humans – an actual 2013. Pathomorphological lesions were health and economic problem. Acta observed in 4 mouflons and 3 deer with Microbiol. Bulg., in press. hyperplastic changes in the small intestine

LABORATORY OF ANTIMICROBIAL AGENTS

Current project

WATER-SOLUBLE PHTHALOCYANINES FOR FLUORESCENCE DIAGNOSIS AND PHOTODYNAMIC THERAPY

Project supervisor: M. Durmuş, PhD, Gebze Institute of Technology, Turkey Project leader: V. Mantareva, PhD, Institute of Organic Chemistry with Centre of Phytochemistry, BAS Project collaborators: The Stephan Angeloff Institute of Microbiology, BAS; Gebze Institute of Technology, Turkey; Institute of Organic Chemistry with Centre of Phytochemistry, BAS Research staff: V. Kussovski, PhD, I. Angelov, PhD Two recently synthesized water- 665 nm). The full photoinactivation was soluble cationic lutetium (III) acetate achieved with 20 μM LuPc-5 for P. phthalocyanines (LuPc-5 and LuPc-6) were aeruginosa and 30 μM LuPc-5 for C. studied as photosensitizers for antimicrobial albicans. The PDI study on microbial photodynamic therapy towards two biofilms showed incomplete pathogenic microorganisms, namely the photoinactivation (<3 logs) for the used Gram-negative bacterium Pseudomonas gentle drug-light protocol. aeruginosa and a fungus Candida albicans. The photodynamic effect was evaluated for Grant No: 212M053 Joint research project the cells in suspensions and organized in 48- TUBITAK (Turkey) – BAS (Bulgaria) h developed biofilms. The relatively high levels of uptakes of LuPc-5 and LuPc-6 References: were determined for fungal cells compared Mantareva, V., Kussovski, V., Durmuş, M., to bacterial cells. The penetration depths and Borisova, E., Angelov, I. (2015). distribution of both LuPcs into microbial Photodynamic inactivation of biofilms were investigated by means of pathogenic species Pseudomonas confocal fluorescence microscopy. The aeruginosa and Candida albicans with photoinactivation efficiency was studied for lutetium (III) acetate phthalocyanines a wide concentration range (0.85–30 μM) of and specific light irradiation. Lasers LuPc-5 and LuPc-6 at a light dose of 50 J Med Sci, in press. cm−2 from red light-emitting diode (LED;

DEPARTMENT OF APPLIED MICROBIOLOGY

LABORATORY OF MICROBIAL BIOSYNTHESIS AND ECOLOGY Final Summary

BIOTECHNOLOICAL AND ECO-FRIENDLY APPROACH FOR EFFECTIVE UTILIZATION OF WASTE PLANT BIOMASS FOR COMPOST AND SOIL IMPROVEMENTS

Project leaders: A. Gousterova, PhD and K. Tsekova, PhD Project collaborators: Inovet OOD, Sofia Research staff: H. Najdenski, DSc; D. Todorova, PhD; D. Paskaleva, MS; I. Tzvetkova, MS During the fulfillment of the biological activity of the ready product activities by the project the main set purpose (compost) and its components, the content was achieved, and namely deriving of cheap of heavy metals and presence of pathogenic and harmless plant bioproducts /compost microflora were researched. and soil improver/ from plant waste. For this In addition during the fulfillment of purpose were made industrial scientific the present project all indicators were researches for developing of methods for achieved, which were set in the contract, and more effective utilization of these waste namely: products, based on enzyme hydrolysis in using of mixed culture of selected Grant BG161РО003-1.1.06 from the termophile microorganisms. The Operational Programme “Development of microbiological and the biochemical the Competitiveness of the Bulgarian processes, which pass during the composting Economics”, EU, European Regional of the plant waste, were studied. The Development Fund.

Current projects

CULTIVATION OF MICRO-ALGAE IN INNOVATIVE PHOTO-BIOREACTOR FOR CARBON DIOXIDE SEQUESTRATION FROM INDUSTRIAL WASTE GASES FROM DIFFERENT SOURCES AND SYNTHESIS OF HIGH VALUE PRODUCTS

Project leader: A. Kroumov, PhD, The Stephan Angeloff Institute of Microbiology, BAS Project collaborators: West Parana State University, Department of Chemical Engineering, Brazil Research staff: A. N. Módenes; D. E. G.Trigueros; F. R. Espinoza-Quiñones; C. E. Borba; F. B. Scheufele; C. L. Hinterholz Theoretical achievements included column photobioreactors (PBRs) modeling, development of the novel strategy for optimization, design and scale-up. The novel

13 strategy was presented in several symposia Preparation of articles on the subject is and congresses and now is in progress of ongoing. publication in Process Biochemistry Journal. Training and supervision of PhD Set up of the Algae Lab. Several student Camila Larissa Hinterholz during stands for performing algal culturing the 3 months period. techniques were set up according to the principles of this strategy. They included Grant "Special Visiting Researcher" under (PBRs) section as follows: section with 16 the program "Science without Borders" small PBRs; section for flat plate PBRs; section for maintaining isolated strains. References: Several algae strains were isolated Kroumov, A.D., Módenesm A.N., from Brazilian lakes and preliminary Triguerosm D.E.G. (2015). A complex morphological identification was performed. theoretical approach for algal medium

Their identification is in progress by using optimization for CO2 fixation from flue modern molecular DNA methods. gas. Acta Microbiol. Bulg. 31(1), 61-70.

EFFICIENT USE OF BIOMASS FOR ENERGY AND ENVIRONMENTAL OBJECTIVES: THE POTENTIAL OF BIOETHANOL AS A FUEL FOR RAW MATERIALS

Project leader: S. D. Ivanova, PhD, Institute of Catalysis, BAS Research staff: A. Kroumov, PhD Project collaborator: The Stephan Angeloff Institute of Microbiology, BAS; Institute of Catalysis, BAS Modeling the process of ethanol finding optimal operating conditions of the production from lignocellulosic wastes process. (LCW), for example, wheat straw. The The development of this model is a model considered simultaneous enzymatic continuation of multiannual work of the hydrolysis of LCW and ethanol synthesis author in the field of process modeling from the hydrolysis products. (laboratory and industrial experiments) of Structure of the model. The author's ethanol production based on different work was focused on enriching our substrates, reflected in his famous 10 knowledge about the process of publications. simultaneous saccharification of lignocellulosic waste (SSLCW) and the Grant E02/1, National Science Fund production of ethanol. The model was used to studying the relationships between state References: parameters and subsequent planning of Kroumov, A.D., Zaharieva, M., Beshkov, V. active experiments. The simulation results (2015). Ethanol from Cellulosic helped to discriminate different working biomass with emphasis of wheat straw hypotheses and accelerate the search of utilization. Analysis of strategies for process development, Int. J. Bioautomat. 19(4), 483-506.

ANAEROBIC BIODEGRADATION OF LIGNOCELLULOSIC WASTE TO PRODUCE

BIOMETHANE AND UTILIZATION OF CO2 FROM IT BY USING MICROALGAE

Project leader: A. D. Kroumov, PhD Theoretical achievements included References: development of the novel strategy for Alexandrov, S., Iliev, I., Gacheva, G., column photobioreactors (PBRs) scheme Kroumov, A., Pilarski, P., Petkov, G. and their modeling, optimization, design and (2015). Review: Could algae be a real scale-up applied for CO fixation from 2 source of fuel? Gen. Plant Physiol. biogas. The purification scheme included two PBRs in series and gas holder where 5(2), 105–122. simulated biogas was loaded. The novel Kroumov, A., Módenes, A., Trigueros, D. strategy was presented in several symposia (2015). A complex theoretical and congresses and now is progress of approach for algal medium publication in Process Biochemistry Journal. optimization for CO2 fixation from flue gas. Acta Microbiol. Bulg. 31(1), Grant DFNI-E01/0001, National Science Fund 61-70.

OPTIMIZATION OF FERMENTATION PROCESSES FOR AMINO ACID PRODUCTION

Project leader: A. Ratkov, PhD Research Staff: I. Dimov, MS; R. Prtkova, MS; F. Filipov, MS; J. Kristeva, MS Caring of fermentation processes for fisico-chemical optimization of the microbial production of different amino processes for their production. acids (mainly lysine, valine and leucine) Bilateral Agreement for Cooperation using different mutant strains as a producers. between The Stephan Angeloff Institute of Optimization of fermentation processes Microbiology, BAS and RTM based on specifics substrate requirement of “Recourses+Technologies Management”, the producers as well as technological and Germany

CARYING OF TEST AND DEMONSTRATION EXPERIMENTS

Project leader: Alexander Ratkov Research Staff: I. Dimov, MS; R. Prtkova, MS; F. Filipov, MS; J. Kristeva, MS Test experiments included evaluation feed grade lysine sulfate as a final product is of specific main raw materials for microbial developed. Education of the Russian staff is production of L-lysine. Testing is focused predicted. on the specificity and the efficiency of Bilateral Agreement for Cooperation Project predicted for industrial production main C- between Institute of Microbiology, BAS and source and main protein source. Based on Company “Plemzavod-Jubileinie”, Russian the specific main raw materials the detailed Federation. technological regalement for production of

LABORATORY OF EXTREMOPHILIC BACTERIA

Current projects

BIODIVERSITY OF HALOPHILIC MICROORGANISMS ISOLATED FROM BULGARIAN SALT NICHES AND THEIR EXOPOLYSACCHARIDE SYNTHESIS POTENTIAL

Project supervisor: M. Kambourova, DSc Project collaborators: University of Food Technologies Research staff: E. Tonkova-Vasileva, PhD; N. Radchenkova, PhD; I. Boyadzhieva, PhD; S. Vasilev, MSc Diversity of halophilic microorganisms 18 substrates were degraded by isolates, in halophilic niches from at the southern with predominance of producers of Black Sea coast was studied by unculturable polygalacturonase, catalase, phytase and and culturable approach. Investigations on lipase. More than a half of the strains (65 %) the archaeal community composition in P18, displayed antimicrobial activity against one the biggest crystallizer pond in Pomorie to five model bacteria and yeasts. salterns (PS) (34% salinity) demonstrated unusually high number of presented taxa in Grant DFNI 02-26/2014, National Science hypersaline environment. Archaeal clones Fund were grouped in 26 different operational taxonomic units (OTUs) assigned to 15 References: different genera from two orders, Halobacteriales and Haloferacales. New Panchev, I., Kuncheva, M., Kovacheva, D., sequences represented 53.9% of archaeal Kamburova, M., Radchenkova, N., OTUs. Present results significantly differed Boyadzhieva, I. (2015). Physical from the previous investigations in regard to characteristics of exopolysaccharide the number of presented genera, the synthesized from halophilic domination of some genera not reported microorganisms of the before in such extreme niche and the Chromohalbactre canadensys strain. identification of previously undiscovered Proceed. Union Bulg. Sci. (Plovdiv) (in 16S rRNA sequences. press) Culturable approach revealed high for Kuncheva, M., Panchev, I., Kamburova, M., such niches taxonomic and metabolic Radchenkova, N., Boyadzhieva, I. diversity: we isolated 19 moderate (2015). Surface active properties of a halophilic and 10 halotolerant strains newly synthesized biopolymer from affiliated with 12 species from eight genera halophilic microorganisms, Scientific referred to the phyla Proteobacteria, works UFT, “Food science, engineering Firmicutes and Actinobacteria. Isolates were and technologies”, Plovdiv, v. LXII, determined as alkalitolerant, and 41 % of 487-489, ISSN 1314-7102. them as psychrotolerant. Half of the tested

CHARACTERIZATION OF ARCHAEAL DIVERSITY IN BULGARIAN HOT SPRINGS

Project supervisor: Assoc. Prof. M. Kambourova, DSc Project collaborator: Department of Biology and Centre for Geobiology, University of Bergen, Bergen, Norway Research staff: N. Boteva, PhD student; I. Boyadzhieva, PhD; N. Radchenkova, PhD; M. Stoilova-Disheva, PhD; D. Lyutskanova, PhD; S. Vasilev, MSc A culture independent approach based one third of them are unclassifiable, as the on the analysis of nearly complete archaeal sequence similarity to the closest classified 16S rRNA genes revealed an unusually high relative in databases is lower than 80%. Low archaeal diversity in one of the hottest similarity with cultured archaeal Bulgarian geothermal springs, Vlasa, representatives and low match to Velingrad (86°C). Archaeal clones were environmental 16S rRNA clones are unique referred to 30 OTUs. They were affiliated features for this thermophilic niche, with both archaeal kingdoms, Euryarchaeota indicating that the archaeal diversity is still and the provisional Proteoarchaeota, with a underexplored. The sequences retrieved predomination of proteoarchaeal clones. from the Vlasa hot spring demonstrate that Representatives of five among seven this environment harbors one of the richest currently provisional proteorchaeal phyla archaeal communities found and contribute were identified in the sample - to expanding ourknowledge of archaeal Crenarchaeota, Thaumarchaeota, diversity in geothermal environments. Aigarchaeota, Korarchaeota, and Geoarchaeota. All sequenced clones EEA Program, Grant D03-100/05.06.2015 showed closest similarity to uncultured representatives from geothermal niches and Reference: some of them formed separate branches. Stefanova, K., Tomova, I., Tomova, A., More than two third of the retrieved Radchenkova, N., Atanassov, I., sequences were >3 % different from their Kambourova, M. (2015). Archaeal and closest matches, suggesting the presence of bacterial diversity in two hot springs representatives of novel species, and even from geothermal regions in Bulgaria as genera in this niche. Four of them differ demostrated by 16S rRNA and GH-57 from the closest relatives with more than genes. Int. Microbiol. (in press) 15%, the border for new phyla. More than New project

EXOPOLYSACCAHARIDE FROM HALOPHILES: PRODUCTION, CHEMICAL CHARACTERIZATION AND THEIR POSSIBLE BIOTECHNOLOGICAL APPLICATIONS

Project supervisor: M. Kambourova, DSc Project collaborator: Institute of Chemistry of Biomolecules, CNR, Naples, Italy Research staff: N. Radchenkova, PhD; I. Boyadzhieva, PhD

LABORATORY OF APPLIED BIOTECHNOLOGIES

Current projects:

RATIONAL PLATFORM FOR SUSTAINABLE PRODUCTION OF PHARMACEUTICALLY RELEVANT MOLECULES FROM PLANTS AND THEIR IN VITRO CULTURE

Project leader: M. I. Georgiev, PhD Project collaborators: Institute of Organic Chemistry with Centre of Phytochemistry-BAS; University of Sofia Research staff: P. Dimitrova, PhD; A. Marchev, PhD; S. Rusinova-Videva, PhD; V. Milanova, Georgi Zahmanov, Elka Genova Chronic (systemic) inflammation and and their in vitro systems. The project is malignant neoplasms are socially-significant built strongly on scientific excellence and diseases, which affect millions of people directly reflects to the National Scientific worldwide. Chemotherapeutic agents Research Strategy and Innovation Strategy currently in use in the clinics did not for Smart Specialization of Bulgaria 2014- succeed in fulfilling their expectations even 2020. though they are very cost-intensive. In Grant DFNI B02/14, National Science Fund parallel, there is an increasing evidence for the potential of plant-derived secondary References: Marchev, A., Yordanova, Z., Alipieva, K., metabolites on the inhibition of different Zahmanov, G., Rusinova-Videva, S., steps of tumorigenesis and associated Kapchina-Toteva, V., Simova, S., inflammatory processes, underlining the Popova, M., Georgiev, M.I. (2015). importance of these products in cancer Genetic transformation of rare prevention and therapy. The aim of the Verbascum eriophorum Godr. plants project is to entail an elaboration of a and metabolic alterations revealed by NMR-based metabolomics. rational platform for sustainable production Biotechnol. Lett. (in press). of pharmaceutically relevant value-added biologically active molecules from plants

RATIONAL PLATFORM FOR HALOGENATION OF HIGH-VALUE PHENYLETHANOID GLYCOSIDES FROM PLANT ROOT CULTURE

Project leader: M.I. Georgiev, PhD Project collaborators: Institute of Botany, Dresden University of Technology (Germany) Research staff: P. Dimitrova; A. Marchev, PhD; S. Rusinova-Videva, PhD The aim of the project of plant biotechnology, molecular biology (EngineeredRoots) is to set to work the and natural products chemistry in order to expertise of the project partners in the field elaborate a technology platform for studying

18 verbascoside (and concomitant strengths of Partners from the EU to deliver phenylethanoid glycosides) halogenation in its objectives for maximum impact. the plant cell/tissue factory. This platform includes the in vitro plant tissue culture Grant DNTS Germany 01/8 (2014-2016) systems (hairy roots), metabolic References: engineering, metabolomics and evaluation Georgiev, M.I., Radziszewska, A., of bioactive properties of the Verbascum’s Neumann, M., Marchev, A., Alipieva, phenylethanoid glycosides. The major K., Ludwig-Müller, J. (2015). Metabolic scientific impact of the project is on alterations of Verbascum nigrum L. strengthening of the knowledge on plants and SAArT transformed roots as bioproduction and engineering of plant- revealed by NMR-based metabolomics. derived high-value metabolites. Plant Cell Tissue Organ Cult. 123, 349- EngineeredRoots is built strongly on 356. scientific excellence and it capitalizes on the New projects

ESTABLISHMENT OF CENTRE OF PLANT SYSTEMS BIOLOGY AND BIOTECHNOLOGY FOR THE TRANSLATION OF FUNDAMENTAL RESEARCH INTO SUSTAINABLE BIO-BASED TECHNOLOGIES IN BULGARIA

Project leader: M. Georgiev, PhD Project collaborators: University of Potsdam (Germany), Max Plank Institute of Molecular Plant Physiology (Germany), Institute of Molecular Biology and Biotechnology (Bulgaria), Maritsa Vegetable Crops Research Institute (Bulgaria) Research staff: Andrey Marchev, PhD The overall objective of PlantaSYST and technological gaps identified in the is to establish a Center of Plant Systems Smart Specialization Strategy of Bulgaria. Biology and Biotechnology (CPSBB) in Besides that, the CPSBB will take a leading Plovdiv, Bulgaria, as a joint initiative of role in educating next-generation early-stage three Bulgarian research institutes located in researchers in the fields of molecular Plovdiv and two institutes located in breeding, plant systems biology and Potsdam, Germany. The ambition of CPSBB biotechnology. is to stand at the forefront of plant sciences Grant 664621, H2020, Teaming Spreading in Bulgaria and Eastern Europe by Excellence integrating molecular biology, functional References: genomics, metabolomics, bioinformatics, Marchev, A., Dinkova-Kostova, A.T., bioprocessing, and long-standing expertise Gyorgy, Z., Mirmazloum, I., Aneva, in practical plant genetics and breeding, to I.Y., Georgiev, M.I. (2015). Rhodiola unravel the plant biology and translating the rosea L.: from golden root to green scientific knowledge into new horticultural cell factories. Phytochem. Rev. (in and industrial applications. Thus, the newly press). established CPSBB will fill in the scientific Skalicka-Wozniak, K., Orhan, E., Georgiev, M.I. (2015). Adulteration of herbal

sexual enhancers and slimmers: the Georgiev, M.I., Xiao, J. (2015). wish for better sexual well-being and Modifications of dietary flavonoids perfect body can be risky. Food Chem. towards improved bioactivity: An Toxicol. (in press). update on structure-activity Chen, L., Teng, H., Xie, Z., Cao, H., relationship. Crit. Rev. Food Sci. Nutr. Cheang, W.S., Skalicka-Wozniak, K., (in press).

Pending support

COMPLEX ASSESSMENT OF DONOR GENOTYPES AND APPROACH FOR CREATION OF BULGARIAN SORTS TOMATOES WITH IMPROVED ANTIOXIDANT CONTENT AND TASTE OF THEIR FRUITS

Project supervisor: B. Bojinov, PhD Project leader: A. Pavlov, DSc. Research staff: I. Ivanov, PhD; T. Gocheva; E. Genova

Grant DFNI-B01/16-1012, National Science Fund

APPLICATION OF OMICS TECHNOLOGIES TO REVEAL HEALTH POTENTIAL OF BULGARIAN HONEY

Project supervisor: M. Shishinjova, PhD Project leader: A. Pavlov, DSc. Research staff: A. Marchev, R. Vrancheva Grant DFNI-B01/31-1012, National Science Fund

RESARCH GROUP OF MATHEMATICAL MODELLING AND COMPUTER METHODS

Current projects

NEW ECOTECHNOLOGIES FOR BIODEGRADATION OF ORGANIC WASTE WITH HYDROGEN AND METHANE PRODUCTION

Project leader: I. Simeonov, PhD Research staff: H. Najdenski, DSc; M. Angelova, DSc; E. Chorukova, PhD; E. Krumova, PhD; V. Kasovski, PhD; L. Kabaivanova, PhD; N. Kostadinova, PhD; S. Mihaylova, MS;V. Hubenov, PhD; L. Dimitrova, PhD student Anaerobic digesdtion of with inclusion of the gas phase; A simplified lignocellulosic wastes was performed with model of the processes of anaerobic optimization to realize higher methane and biodegradation of organic waste with the hydrogen production. Chemical and production of hydrogen and methane; A biological (rep Basidiomycota) methods for model of the processes for anaerobic pretreatment of wheat straw were tested. biodegradation of organic waste to obtain Comparative studies aimed at obtaining hydrogen and methane based on ADM1; biogas from untreated and treated wheat Method for decontamination of compost straw in laboratory bioreactors were with was studied. Studies on the process of conducted. Comparative studies on the production of hydrogen from substrate production of biogas from untreated wheat cellulose were initiated. and barley straw in a pilot bioreactor were also performed. Mathematical models were Grant DFNI-E02/13, National Scientific developed for the following processes: AD Fund

ERASMUS+ (FRANCE)

Project leader: I.Simeonov, PhD Teaching activity: I. Simeonov, PhD Content of the teaching programme: 3. Optimisation and control of 1. Mathematical modeling of biotechnological processes biotechnological processes 2. Parameters and states estimation Grant: European Union, Erasmus+

DEPARTMENT OF VIROLOGY

Current Projects:

A NOVEL APPROACH TO HIGHLY EFFICIENT CHEMOTHERAPY OF ENTEROVIRAL INFECTIONS

Project leader: A.S. Galabov, DSc Project collaborator: M. Arita, PhD; National Institute of Infectious Diseases, Gakuen, Tokyo Research staff: I. Nikolova, PhD; A. Stoyanova, MS; P. Grozdanov, PhD; N. Petrov, PhD; Y. Abashev, PhD; S. Philipov, PhD; N. Vilhelmova-Ilieva, PhD; L. Mukova, MS; N. Nikolova, MS; P. Stoyanova, DVM; Nikolaeva-Glomb, MD, PhD; I. Zahova; E. Dimitrova Chemotherapy could be a leading tool resistance, a phenomenon of increased drug for controlling enterovirus (EV) infections, sensitivity was established. Pleconaril but clinically effective anti-EV drugs do not sensitivity increased by 7 times on Day 7 currently exist, mainly due to the and by over 49 times on Day 13; MDL-860 development of drug resistance. We were sensitivity increased by 29 times on Day 7, the first to investigate the combination and oxoglaucine sensitivity by 6 times on effects of selective EV replication inhibitors Day 13. Daily, simultaneous administration in order to limit this process. In previous of PMO showed no protective effect and a studies, we showed the efficacy of rapid development of drug resistance. consecutive alternating administration These results add new support for (CAA) of the triple combinations using CAA treatment courses to achieve disoxaril/guanidine/oxoglaucine and clinically effective chemotherapy of EV pleconaril/guanidine/ oxoglaucine against infections. Coxsackievirus B1 (CVB1) infection in newborn mice. Drug sensitivity tests of the Grant Б-01-13/12, National Science Fund viral brain isolates showed that these drug combinations prevented the development of References: drug resistance. Galabov, A. S., Nikolova, I., Vassileva, R., In the current study, we replaced Stoyanova, A. (2015). Antiviral guanidine-HCl with enteroviral RNA combination approach: a perspective synthesis inhibitor MDL-860 to test the to combat enterovirus infections. - effect of a new triple combination - Prilozi/Contributions, 36, 91-99. pleconaril/MDL-860/oxoglaucine (PMO) - Arita, M., Philipov, S., Galabov, A.S. applied via CAA in newborn mice infected (2015). Phosphatidylinositol 4-kinase subcutaneously with 20 MLD50 of CVB1. III beta is the target of oxoglaucine The PMO combination via CAA showed and pachydipol (Ro 09-0179) for their high activity at the 75 mg/kg MDL-860 anti-poliovirus activity, and locates at dose: a protective effect of 50% and a upstream of the target step of brefeldin pronounced suppression of brain virus titers. A in poliovirus replication. Microbiol. Moreover, along with the prevention of drug Immunol. 59, 338-347.

Stoyanova, A., Nikolova, I., Galabov, A.S. Stoyanova, A., Galabov, A. S. (2015). (2015). Effect of consecutive Enteroviruses and perspectives for alternative administration (CAA) of a etiotropic therapy of enteroviral triple anti-enteroviral combination on infections. Acta Microbiol. Bulg. 31, Coxsackievirus B1 neuroinfection in 93-106. mice. Antivir. Res. 121, 138-144.

SYNTHESIS AND ANTI-ENTEROVIRUS ACTIVITY OF NOVEL DIARYL ETHERS AND THEIR COMPLEXES WITH CYCLODEXTRINS

Project supervisor: L. Antonov, DSc, Institute of Organic Chemistry with Center for Cytochemistry, BAS Project leader: A. S. Galabov, DSc Project collaborator: Institute of Organic Chemistry with Center for Cytochemistry, The Stephan Angeloff Institute of Microbiology, BAS Research staff: V. Dimitrov, DSc; G. Dobrikov, PhD; P I. Nikolova, PhD; A. Stoyanova, MS; L. Mukova, MS,; N. Nikolova, MS; P. Grozdanov, PhD The synthesis of a series of more than (Sabin strain) (PV1), Coxsackievirus B1 60 new diaryl ether compounds was (Connectucut strain) (CVB1) and accomplished starting from 2-chloro-5- Coxsackievirus B3 (Nancy strain) (CVB3). nitrobenzonitrile and diffferent phenols. The The results obtained at the testing of the first reactions were carried out under basic series of 25 compounds manifested a conditions (NaOH or KOH) in DMSO. Most pronounced effect of VGA-10-2 vs. PV1 of the compounds are near analogues of and CVB1 (SI of 118, and 405, DNB – well known active compound against respectively). Two compounds, CB-64 and enteroviruses. All compounds were obtained CB-69, showed a moderate activity (SI in high yields and purity after column around 20) vs. CVB3. The starting chromatography and were characterized by compound DNB demonstrated a pronounced NMR, MS-spectra, melting points and inhibitory effect against the three tested elemental analysis. viruses - SI of 72.5, 586.9 and 182, The new synthesized diarylethers were respectively. tested for their anti-enteroviral activity against three enteroviruses: poliovirus 1 Grant Б 02-11/12.12.2014, National Science Fund

A MODERN ALTERNATIVE FOR THE PROPHYLAXIS AND TREATMENT OF INFLUENZA VIRUS INFECTION – MULTITARGET APPROACHES WITH HIGHLY EFFICIENT COMBINATIONS OF ANTIVIRAL CHEMOTHERAPEUTICS AND BIOLOGICALLY ACTIVE COMPOUNDS

Project leader: L. Simeonova, PhD Project collaborator: Sofia University “St. Kliment Ohridski” Research staff: E. Pavlova, PhD; L. Tancheva, PhD; G. Goujgoulova, PhD; I. Slavcheva, MS; N. Zografov, MS; G. Gegova, MS; L. Mukova, MS; K. Todorova, BS; S. Andreeva; N. Petrov, PhD Advisors: Acad. A. S. Galabov, DM, DSc; G. Georgiev, DVM, DSc; V. Savov, DSc; M. Mileva, PhD The necessity of efficient alternative enzymatic activity and was found that the approaches in antiinfluenza therapy oxidative levels were significantly reduced continues to exist since novel antiviral for combinations-treated groups, the effects inhibitors are still in an experimental phase of compounds on hepatic drug-metabolizing of their development. As a strategy to systems were studied. combat the severity of the disease a complex Further study: Individual and combined inhibition of different aspects in the effects of oseltamivir and and siRNAs in pathogenesis of flu could be considered, i.e. cell cultures against seasonal influenza applying combinations of the available anti- strains: H3N2, H1N1v; influenza inhibitors and potent biological Final experiments were carried out on response modifiers. Beyond the cell the combination effect of α-tocopherol and disruptions caused by the viral replication oseltamivir. Strongly dose-dependent itself, influenza infection is associated with augmented antiviral effect of the a dramatic increase in the amount of free combination α-tocopherol and 0.625 mg/kg radicals in the organism, leading to a state of oseltamivir was demonstrated when α- oxidative stress. Viral activation of tocopherol was administered simultaneously phagocytes generates free radicals and with oseltamivir: a pronounced decrease in active forms of oxygen severely damaging mortality rate (a 78% protection), and a cellular membranes. lengthening of mean survival time by 3.2–4 The hypothesis of efficacy of a days. Lung parameters showed a substantial combined multitarget approach was tested decrease in infectious virus content (∆ logs by standard virological, pharmacological = 3.8/4.1) and a marked diminishment of and biophysical methods in vitro and in vivo. lung index and pathology. Combination α- For the implementation of the project the tocopherol with 1.25 mg/kg oseltamivir following activities were performed: manifested a marked protective effect, but Antioxidant capacities of oseltamivir, ellagic the effect on lung parameters was less. The acid, isprinosine were tested in cell-free combination effect of α-tocopherol with 2.5 model systems; toxicity of ellagic acid and mg/kg oseltamivir did not surpass the protective effects of oseltamivir combined monotherapeutic effect of oseltamivir. When with antioxidants and immune modulator in α-tocopherol was applied in courses starting mice against experimental H3N2 infection 5 or 2 days before infection, its combination were established; levels of oxidative stress with oseltamivir was ineffective. in experimentally infected and treated mice were investigated by measurement of Grant B-01-19/12, National Science Fund

BALKAN ENDEMIC NEPHROPATHY

Project leaders: A. S. Galabov, DSc and. M. Polenakovic, MD, DSc, Macedonian Academy of Sciences and Arts (MASA), Skopje, FYROM Project collaborators: Department of Medical Genetics, Medical University of Sofia, Macedonian Academy of Sciences and Arts (MASA), Skopje, FYROM Research Staff: D. Toncheva, DSc and collaborators; P. Grozdanov, PhD; I. Nikolova, PhD The main work on the project in 2015 Staneva, R. G., Balabanski, L., Dimova, I., was the preparation of the manuscript of the Rukova, B. B., Hadjidekova, S., monograph “Balkan Endemic Nephropathies Dimitrov, P., Simeonov, V., Ivanov, - World Wide Diseases”. Authors of the S., Vagarova, R., Malinov, M., book are from Bulgaria, Serbia and Republic Cukuranovic, R., Stefanovic, V., of Macedonis. Editor-in-Chief is Polenakovic, M., Djonov, V., academician Vladislav Stefanovic from Galabov, A., Toncheva, D. I. (2015). Serbian Academy of Sciences and Arts, Genetic, genomic and epigenomic Skopje, FYROM. studies of Balkan Endemic Academician Angel S. Galabov and Nephropathy (BEN). Prilozi/ Corr.-Member Draga Toncheva are among Contributions, 36, 101-108. the editors. They are authors of four chapters Stefanovic, V., Toncheva, D., Polenakovic, of the monograph: „Role of Viruses in M. (2015). Balkan nephropathy. Clin. BEN” (A. S. Galabov; co-authors of the Nephrol. 83 (7 Suppl.), 64-69. chapter B. Kamarinchev and T. Stoitsova ), Popovska-Jankovic, К., Noveski, P., „Immunologic Mechanisms” (А. S. Galabov Jankovic-Velickovic, L., Stojnev, S., and A. Pashov), „Genetics of BEN” (D. Cukuranovic, R., Stefanovic, V., Toncheva and co-authors.) and “Epigenetics Toncheva, D., Polenakovic, M., of BEN” (D. Toncheva and R. Staneva). Plaseska-Karanfilska, D. (2015). Bilateral joint project (2014-2015) in the MicroRNA profiling in patients with frame of agreement between Bulgarian upper urothelial carcinoma associated Academy of Sciences and Macedonian to Balkan endemic nephropathy Academy of Sciences and Arts (BEN), BioMed Res. Internat. supplied.

References:

BALKAN ENDEMIC NEPHROPATHY

Project leaders: A. S. Galabov, DSc and V. Stefanovic, MD, PhD, Serbian Academy of Sciences and Arts (SASA) Project collaborators: Department of Medical Genetics, Medical University of Sofia, Serbian Academy of Sciences and Arts (SASA), Serbia Research staff: D. Toncheva, MD, DSc and collaborators; P. Grozdanov, PhD; I. Nikolova, PhD Internet base data for patients with on BEN patients for genomic and proteomic Balkan endemic nephroparthy (BEN) is analyses was broaden. DNA from 22 established, containing detailed clinical and patients was isolated. Ekzome analysis by pathological characteristics. The DNA bank NGS of 22 000 genes was done. Some

25 specific and general changes in BEN Role of viruses in BEN – Angel associated copies were identified. Galabov The main work on the project in 2015 Immunologic mechanisms – Angel was the preparation of the manuscript of the Galabov, Anastas Pashov monograph “Balkan Endemic Nephropathies Proteomics of BEN – Vladislav - World Wide Diseases”. Authors of the Stefanovic, Ivana Markovic, Hasan book are from Bulgaria, Serbia and Republic Dihazi, Guerhard Mueller of Macedonis. Editor-in-Chief is Genetics of BEN – Draga Toncheva et academician Vladislav Stefanovic from al Serbian Academy of Sciences and Arts, Epigenetics of BEN - Draga Serbia. Toncheva, Rada Staneva Interacademic Bilateral Collaborative Pathology of BEN – Ljubinka Project BAS - Serbian Academy of Sciences Jankovic Velickovic and Arts (2014-2015) Urithelial cancer – Vladislav Stefanovic, Zoran Radovanovic References: Biomarkers of BEN and urothelial Balkan Endemic Nephropathies - World cancer – Liubinka Jankovic Wide Diseases. Academician Angel S. Velickovic, Vladislav Stefanovic Galabov and Corr.-Member Draga Diagnosis of BEN – Vladislav Toncheva are among the editor (see Stefanovic, Momir Polenakovic previous project). Screening of BEN – Vladislav BEN history – Zoran Radovanovich, Stefanovic, Momir Polenakovic Tsvetan Dimitrov Prevention and treatment of BEN – Epidemiology of BEN – Zoran Vladislav Stefanovic, Momir Radovanovich, Tsvetan Dimitrov Polenakovic Etiology of BEN – Vladiav Endemic Nephropathy beyond the Stefanovic, Momir Polenakovic, Balkans - Momir Polenakovic, Draga Toncheva et al. Vladislav Stefanovic Environmental toxins (Aristolochia, Further research of BEN and mycotoxins, polycyclic aromatic associated urothelial cancer. What has amines, others) – Vladislav to be done - Vladislav Stefanovic, Stefanovic, Draga Toncheva et al. Draga Toncheva

THE PRIMARY LIVER CANCER IN BULGARIA: HEPATITIS VIRUSES AND ENVIRONMENTAL FACTORS

Project leaders: A. S. Galabov, DSc and P. Pineau, PhD, Institut Pasteur, Paris Research staff: L. Doumanova, PhD; P. Grozdanov, PhD Research partners: R. Gaydarski, DSc; Tokuda Hospital Sofia; V. Dimitrova, MD, DSc, Clinics of Surgery, Alexandrovska Hospital, Sofia The collecting of samples from biopsy, in parallel with blood samples was patients with primary liver cancers treated continued. surgically or undergoing the needle liver ACIP Project

GENETIC PROFILE OF THE POPULATION OF BULGARIAN LANDS DURING THE THYRACIAN PERIOD: NEW PERSPECTIVES ON BULGARIAN HISTORY

Project supervizor: A. S. Galabov, DSc Project supervisor: D. Toncheva, MD, DSc, Department of Medical Genetics, Medical University of Sofia Project collaborator: D. Caramelli, PhD, University of Florence, Italy, Department of Anthropology Research staff: D. Nesheva, PhD student; S. Karachanak-Yankova, PhD; P. Grozdanov, PhD; I. Nikolova, PhD Analysis of antic DNA restored from References: bone residues discovered from tumuli dating Nesheva, D. V., Karachanak-Yankova, S., from the Thracian period is done for the first Lari, M., Yordanov, Y., Galabov, time. Genetic origin of the population of A.S., Caramelli, D., Toncheva, D. Bulgarian lands from this period is (2015). Mitochondrial DNA Suggests determined and the contribution of this a Western Eurasian origin for Ancient population to the contemporary Bulgarian (Proto-) Bulgarians. Human Biology, genetic fond is estimated. 80 samples from Open Access Pre-Prints. Paper 69, tumuli of the 3th millennium before Christ http://digitalcommonswayne.edu/hum in the regions of Stara Zagora and Sliven bici were studied. Isolated mtDNA Karachanak-Yankova, S., Nesheva D., (hipervariable segments I) were sequenced Galabov, A. S., Toncheva, D. (2015). by the classic sequencing and the NGS Distribution of East Eurasian Y- analysis. The results obtained were chromosome and mitochondrial DNA compared to the respective areas of mtDNA haplogroups across Eurasia: insights of contemporary Bulgarians. Genetic into the genetic ancestry of profiles were compared in historical aspects. Bulgarians. Adv. Anthropol. 5, 205- 266.

MODIFYERS OF BIOLOGICAL REPLY AT INFLUENZA INFECTION

Supervizors: M. Mileva, PhD and A.S. Galabov, DSc, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences; D. Krastev, DSc, Medical University of Sofia PhD Student: A. Dimitrova, PhD student The studies carried out show that S- dose (applied at 2.5mg/kg daily in two adenosyl-L-methionine (SAM) has a intakes for 5 days, starting from 4 h before protective effect in a model of influenza virus inoculation) in infected with influenza infection in mice. This substance converts in A virus mice. SAM was. Survival was 70% glutathione - the main antioxidant in the in the treatment with oseltamivir and raised body, through multistep biochemical cycle. to 90% in the treatment with oseltamivir and The effect of combined treatment with SAM SAM in both of doses. SAM alone does not (given as a single daily dose of 50 and 100 show any antiviral activity. The present mg/kg in different mice groups starting from findings suggest that therapy with molecules 5 days before infection until day 4 after converted in antioxidants in the body infection) and the antiviral agent oseltamivir increases survival by modulating the host

27 defense mechanisms and by a direct References: antioxidant effect against oxidative stress Dimitrova, A., Mileva, M., Krustev, D., associated with viral infections. This study Galabov, A.S. (2015). Some features in experimental influenza viral demonstrated the effectiveness of combining infection in mice and adequate agents that act through different mechanisms markers used for their assaying. In: - antiviral drug oseltamivir as specific Proceedings of the Sixth Workshop on neuraminidase inhibitor of influenza virus, Experimental Models and Methods in and SAM as precursor of most important Biomedical Research, IEMPAM, antioxidant - glutathione. Sofia, 2-14 May 2015, pp. 136-143.

DEPARTMENT OF IMMUNOLOGY

Final Summary

FUNCTIONAL ELIMINATION OF AUTOREACTIVE T CELLS BY ANTIBODY THERAPY IN MURINE MODEL OF SYSTEMIC LUPUS ERYTHEMATOSUS

Project leader: A. Tchorbanov, PhD Project collaborator: J. Prechl, PhD and M. Herbáth, PhD, Immunology Research Group, Hungarian Academy of Sciences, Eötvös Loránd University Research staff: N. Mihaylova, PhD; N. Kerekov, PhD student; S. Chausheva, graduate student; Silvya Bradyanova, graduate student; Petroslav Chipinski, graduate student Systemic lupus erythematosus (SLE) neutralizing monoclonal antibody against is the prototype systemic autoimmune the AnxA1. disease, characterized by the generation of The immunomodulatory activity of autoantibodies specific for native (ds)DNA monoclonal anti-AnxA1 antibody was tested and nucleic acid–protein complexes, the on an in vivo experimental model of human formation of immune depositions, and SLE. Six to eight week-old immunodeficient inflammation in different organs and tissues. SCID mice were reconstituted by In recent years, SLE research has focused on intraperitoneal (i.p.) injection of PBMCs the role of B and CD4+ T-cells in disease from clinically relevant SLE patients. The pathogenesis. Current therapies of the control group of SCID mice was injected disease are non-specific and often intraperitoneally with PBMCs from healthy counterbalanced by adverse effects. They volunteers. The humanized mice have been are mainly based on immunosuppressive treated at weekly intervals with the drugs such as corticosteroids and monoclonal anti-AnxA1 antibody. All mice cyclophosphamide, which are administrated were bled weekly and collected sera were at high doses in phases of exacerbation. kept at -70ºC. Annexin A1 (AnxA1) (37 kDa), was The changes of human cell originally identified as a phospholipase A2 populations after transfer in SCID mice with (PLA2)-inhibitory protein and second or without treatment with anti-AnxA1 messenger of glucocorticoid antibodies were analyzed by flow cytometry pharmacological effects. Analysis of AnxA1 (CD3, CD4, CD8, CD19, CD25, CD69). expression in T cells from patients suffering The kinetics of anti-DNA human IgG from rheumatoid arthritis showed higher antibodies as well as the other auto-antibody levels of this protein compared to healthy panels and the cytokine secretion (IL-4, control volunteers, providing clinical IL10 and IFN-γ) were determined by relevance to the role that AnxA1 might play ultrasensitive ELISA. The glomerular in autoimmune diseases. We hypothesize deposition of human IgG were studied in that it may be possible to down-regulate the selected animals. The degree of albumiuria activity of autoreactive T and B cells from was regularly measured. The life-span of the lupus patients by treating them with a treated mice was prolonged compared to that of the controls.

Bilateral grant between Bulgarian Academy Sciences of Sciences and Hungarian Academy of

Current projects

ANTIBODIES WITH INDUCED POLYREACTIVITY – ROLE IN IMMUNE HOMEOSTASIS AND THERAPEUTIC POTENTIAL

Project leader: Tch. Vassilev, DSc Research staff: A. Pashov, PhD; I. Djoumerska-Alexieva, PhD; M. Hadzhieva, PhD student; J. Dimitrov, PhD. Antibody repertoires of healthy heme-exposed pooled human IgG in the humans and animals contain a fraction of same models of sepsis and aseptic systemic antibodies able to acquire additional inflammation. The administration of a single polyspecificity following exposure to dose of heme-exposed pooled IgG resulted several biologically relevant redox in a significant increase in the survival of molecules (free heme, reactive oxygen mice with endotoxinemia, but not in those species (ROS), ferrous ions, HOCl, etc.). with polymicrobial sepsis and zymosan- The physiological role of these "hidden" induced severe generalized inflammation. polyspecific antibodies is poorly understood. Antibodies with induced polypecificity Similarly to inherently polyspecific acqure novel biological properties and antibodies, those with induced therapeutic potential, however these depend polyspecificicty may also have on the mechanisms for modification used. immunoregulatory properties. We have previously shown that a pooled human IgG Grant DFNI-B01-29, National Science Fund preparation, modified by the exposure of ferrous ions, acquires the ability to References: significantly improve survival of animals Hadzhieva, M, Vassilev, T, Bayry, J, with polymicrobial sepsis or aseptic Kaveri, S, Lacroix-Desmazes, S, systemic inflammation induced by bacterial Dimitrov, J.D. (2015) Relationship lipopolysaccharide or zymosan between natural and heme-mediated administration. In the present study we have antibody polyreactivity. Biochem. analysed the effects of administration of Biophys. Res. Comm. (in press).

DESIGN OF MODIFIED IMMUNOGLOBUIN PREPARATIONS WITH ADDITIONALLY INDUCED POLYSPECIFICITY FOR PASSIVE IMMUNOTHERAPY OF SEPSIS (BULGARIAN/SWISS COLLABORATIVE PROJECT)

Project leader: Tch. Vassilev, MD, DSc Project colaborators: S. von Gunten, MD, PhD; C. Schneider, PhD student, University of Bern, Bern, Switzerland Research staff: М. Hadzhieva, PhD student; A. Pashov, PhD; I. Djoumerska-Alexieva, PhD; J. Dimitrov, PhD; N. Bovin, PhD;, S. von Gunten, PhD On the basis of preliminary data, we in all three models. The studies of the have hypothesized that the passive mechanisms of beneficial action of the latter immunotherapy with pooled preparation revealed its ability to bind to immunoglobulin preparations with proinflammatory molecules, complement additionally enhanced polyspecificity could components and extracellular histones. neutralize some of the products of the genomic storm and thus should be beneficial Grant by the Swiss National Science fund in systemic inflammatory syndromes, and the Bulgarian Ministry of Education and regardless of their primary insult. Three Science (IZEBZO) models of systemic inflammation in the presence or absence of infection were used References: Djoumerska-Alexieva, I., Roumenina, L., to check this hypothesis: induced by LPS, induced by zymosan and induced by cecal Pashov, A., et al. (2015). Intravenous ligation and puncture (CLP). Although the immunoglobulin with enhanced infusion of native IVIg had no effect on polyspecificity improves survival in survival, the administration of the same experimental sepsis and aseptic single dose of the Fe(II)-exposed IVIg systemic inflammatory response significantly improved the survival of mice syndromes. Mol. Med. 21, 1002-1010.

STUDIES OF IMMUNOSIGNATURES IN THE ANTIBODY REPERTOIRES OF PATIENTS WITH PRIMARY MALIGNANT AND METASTATIC BRAIN TUMORS

Project leader: A. Bussarski, PhD Project collaborator: Neurosurgery Clinic of the St. Ivan Rilsky Hospital of the Medical University in Sofia, Bulgaria Research Staff: A. D. Pashov, PhD; D. Ferdinandov, MD; V. Kostov, student The laboratory partnered in this project of binding to a large set of peptides from funded by an intramural microgrant from the tumor antigens. The grant also covered the Medical University. This collaborative preliminary test with 3 tumor antigen project aimed at the building of a collection microarray chips produced by the of sera from patients with glioblastoma PepperPrint, Heidelberg. multiforme, lung cancer brain metastases or non-tumor bearing surgery patients to probe Grant from the Medical University. The the capacity of serum IgM reactivity to collection of sera is available and the predict the diagnoses based on the patterns analysis of 21 patients was further funded

31 and carried out in the context of the BG-09 DFNI-B01-29, Bulgarian Science Fund. D03-103 EEA grant and, in part, by the

RATIONAL DESIGN OF ANTIBODY REPERTOIRE PROBES USING PEPTIDE ARRAYS

Project leader: A. D. Pashov, PhD Project collaborator: E. Hovig, PhD; T. Clancy, PhD, Oslo University Hospital, Oslo, Norway Research Staff: Tch. Vassilev, DSc; M. Hadzhieva, PhD student, V. Shivarov, postdoc; V. Kostov, student During the first year of the project data designed and assembled together in a on the IgM reactivity of the sera from 21 pipeline for the automation of the analysis. patients with glioblastoma multiforme, lung In the framework of the BG09 D03- cancer brain metastases or non-tumor 103 grant the Laboratory organized an bearing surgery patients was analyzed to international conference “The Antibody determine the capacity of the IgM antibody Repertoire as a Biomarker”, BAS, Sofia, repertoire dynamics to predict the diagnoses 23.09.2015 based on the patterns of binding to a large (https://www.facebook.com/EAEconference set of peptides from tumor antigens. 203 ARB/?ref=aymt_homepage_panel ) with the peptide reactivities differed significantly participation of vaccinologists, between the groups and could be used to immunologists and bioinformatitions from classify the patients correctly (“leave on USA, France, Switzerland, Norway and out” validation – 100%). Most of the work Russia. during this period was dedicated to adopting the appropriate bioinformatics methodology Grant BG09 D03-103, National Science for the analysis of peptide array probes of Fund antibody reactivity. A number of tools were

STUDY ON THE IMMUNE MODULATION OF DISEASE SEVERITY IN HUMANIZED SCID MURINE MODEL OF IMMUNOLOGICAL DISEASES

Project leader: A. Tchorbanov, PhD Project collaborator: B.-L. Chiang, PhD; K.-H. Chu, PhD; C.-J. Chiu, PhD student; Graduate Institute of Immunology, National Taiwan University, Taiwan Research staff: N. Mihaylova, PhD; K. Nikolova-Ganeva, PhD; I. Manoylov, PhD student Systemic lupus erythematosus is a that a down-modulation of activated self- polygenic pathological disorder with reactive T and B cells involved in the multiple organ involvement. Self-specific B pathogenic immune response is a reasonable cells play a main role in the lupus approach for SLE therapy. pathogenesis by generation of Allergic diseases have been increasing autoantibodies as well as by serving as in both developed and developing countries important autoantigen-presenting cells. in the past decades. Allergic diseases such as Autoreactive T lymphocytes, on the other bronchial asthma, allergic rhinitis and atopic hand, are responsible for B cell activation dermatitis have been related to T helper cells and proliferation, and cytokine production. function. Certain cytokines such as IL-4, IL- Therefore, both evidences promote the idea 5, IL-13 have been found to play the critical

32 role in the pathogenesis of allergic diseases. possibility on application of Treg cells, For the treatment of allergic diseases, especially Treg/B cells for the treatment of conventional therapeutic approaches immunological diseases. including anti-histamine and steroid, novel Characterization of regulatory T cells treatments are still needed for these allergic induced by B cells for the treatment of diseases. autoimmune diseases. B cells isolated from Most recently, we identified a Peyer’s patches and spleens were cultured particular subset of regulatory T cells with OVA and CD4+ CD25- T cells. After 3 induced by B cells (Treg/B cells), which days, B cell-primed T cells (Treg-of-B cells) could be applied for the future treatment of were harvested. The cytokine profile and autoimmune or allergic diseases. In this suppressive function were assayed to collaborative project, we aim to study the characterize these Treg-of-B cells. role of regulatory T cells in the pathogenesis of both autoimmune and allergic diseases. In Bilateral grant between Bulgarian Academy addition, we also like to explore the of Sciences and Ministry of Science and Technology, Taiwan.

SECTION OF MYCOLOGY

Final Summary:

STUDY OF THE PHYSIOLOGICAL, BIOCHEMICAL AND ECOLOGICAL CHARACTERISTICS OF MICROMYCETES RESISTANT TO HEAVY METAL STRESS

Project leader: M. Angelova, DSc Project collaborator: V. Grishko, PhD, Ukrainian Academy of Sciences, Botanical Garden Research staff: S. Pashova, MS; R. Abrashev, PhD; E. Krumova, PhD; N. Kostadinova, PhD; J. Miteva-Staleva, MS; B. Spassova, MS; E. Evtova. Contamination of the environment by metal ions seems to be strain-dependent. heavy metals is one of today’s serious Both strains, Aspergillus fumigatus 32 and problems. A number of industrial and Aspergillus fumigatus G exhibit varying agricultural activities cause a daily increase degrees of damage from oxidative stress and in the concentration of toxic waste in the soil different strategy of antioxidant protection. and water. At the same time, heavy metals More clear response against metal-induced are associated with damages to human stress has been shown at the strain isolated health. .The aim of the project was to from Bulgarian contaminated soil. investigate the relationship between oxidative stress and heavy metal exposure in Bilateral joint project (2013-2015) in the resistant fungal strains and the role of frame of agreement between Bulgarian antioxidant defence in this resistance. The Academy of Sciences and National experiments were performed with two Academy of Sciences of Ukraine. fungal strains Aspergillus fumigatus, isolated from contaminated soils in Bulgaria References: and Ukraine. A study on the adaptive Krumova, E., Miteva-Staleva, J., response against copper, zinc, nickel and Kostadinova, N., Korinovska, O.M., cadmium mixture as a admissible Abrashev, R., Spasova, B., Angelova, concentration limit (ACL) 0.75, 1.0, 2.0, 5.0 M. (2015). Еffect of cadmium ions on was carried out. The heavy metal effect was the fungal strain Aspergillus fumigatus evaluated by the changes in fungal growth 32 isolated frommetal polluted soil. and metal ion accumulation, the level of Acta Microbiol. Bulg. 30, 1, 40-47. oxidative stress biomarkers, and the Grshko, V., Korinovska, O.M., Angelova, antioxidant activities of enzymes such as M., Krumova, E. (2015). Sensivity to superoxide dismutase (SOD) and catalase heavy metals compounds of some (CAT). The strain isolated from Ukraine has micromycetes isolated from soils of a higher threshold for the accumulation of Ukraine and Bulgaria. Biology and metal ions in the cells. The adaptive Ecology Hruntyv, Lvov, 14-16 October, response involved clearly prononced 20-21. oxidative stress events. The model strains Krumova, E., Kostadinova, N., Miteva- demonstrated increase in oxidative damaged Staleva, J., Gryshko, V., Angelova, M. proteins content and reserve carbohydrates (2015). Cell response to Cu- and Zn- level. In addition, activation of antioxidant induced oxidative stress in Aspergillus enzyme defence was evaluated. The fumigatus isolated from polluted soils. tolerance to different concentrations of CLEAN – Soil, Air, Water, (in press).

EFFICACY OF ANTIMICROBIAL SUBSTANCES PRODUCED BY STREPTOMYCES IN MODIFYING POLYMERS USED IN CONSERVATION OF PAINTINGS IN SOME ANCIENT EGYPTIAN TOMBS

Project leader: M. Angelova, DSc Project collaborator: M. Farrag, PhD, Zagazig University, Faculty of Science, Botany Department, Egypt Research staff: S. Pashova, MS; R. Abrashev, PhD; E. Krumova, PhD; N. Kostadinova, PhD; J. Miteva-Staleva, MS; B. Spassova, MS; E. Evtova Ancient Egypt is known for its by classical taxonomy methods as invaluable culture monuments. Egyptian filamentous fungi belonging to the phyla pyramids and tombs are a part of the world Ascomycota and Basidiomycota. heritage. At the same time, they are exposed Representatives of each morphology group to various factors (physical, chemical and and from each sample were successfully biological) that cause deterioration and identified to species or genus level by degradation. Mural paintings and stone sequencing analysis of ITS regions and the surfaces in the tombs suffer from different SSU ribosomal RNA gene. The results deterioration symptoms such as confirmed the affiliation of all isolates to the discoloration, structure alterations, and subkingdom Dikarya. Ten fungal strains microbial penetration into painting layers. belonged to Penicillium genus, four strains Filamentous fungi play an important role in to Aspergillus genus, one strain to biodeterioration of these objects. Cladosporium genus and one strain to The aim of the present project was to Bjerkandera adusta species. Sequence describe the fungal contamination in ten comparison with the GenBank data (NCBI) Egypt tombs and mosques as a first step in showed over than 99 % identity with development of appropriate conservation sequences from type strains or culture method. For this purpose, molecular collection strains. identification of the isolated fungal strains was performed. Also, the relation between Bilateral scientific cooperation of the composition of limestone support and fungal Bulgarian Academy of Sciences and the growth was proven. Zagazig University, Egypt. The study provides new data of the harmful inhabitants of the Egypt tombs. A References: total of 30 isolates were obtained from the Stoyancheva, G., Krumova, E., analyses of 13 samples taken from different Kostadinova, N., Miteva-Staleva, J., tombs in Egypt through soil dilution and soil Grozdanov, P., Ghaly, M. F., Sakr, A. sprinkle plates techniques. All the strains A., Angelova, M. (2015). Phylogenetic were isolated from agar plates incubated at identification of filamentous fungi 28°C. The isolates were obtained in pure isolated from ancient Egypt tombs. cultures by single conidial transfer onto beer Fungal Biol. (submmited). agar plates. They were grouped and defined

Current projects

FUNGI AS SOURCE OF LIGNOLYTIC ENZYMES. PRODUCTION AND POSSIBLE APPLICATIONS.

Project leader: E. Krumova, PhD, Project collaborator: E. Savino, Departament of Ecology and Land and Terrestrial Environments, University of Pavia, Italy Research staff: M. Angelova, DSc; R. Abrashev, PhD; V. Dishliiska, MS; J. Miteva-Staleva, MS; N. Kostadinova, MS; S. Vassilev, MS; B. Spassova, MS; E. Evtova. The study of wood decay macrofungi the capacity to secrete cell wall degrading that naturally biodegrade lignocellulosic enzymes growing on poplar wood chips and polymers has been steadily increasing due to sawdust. We tested four lignicolous species their numerous and innovative applications. ( Daedalea quercina, Fistulina hepatica, In the last decade, the researchers of Lenzites warnieri, Schizophyllum commune ) Mycological Laboratory of Pavia University as regards their antioxidant activity. The two (Italy) and of The Staphan Angeloff Institute species poorly or not investigated till now of Microbiology, BAS isolated in pure for this activity, D. quercina and L. culture wood decay macrofungi collected in warnieri , revealed the most interesting their own country. A few species have been results. investigated as regards different aspects: the growth profile at different temperature and Italian-Bulgarian bilateral project (2014- 2018)

LIST OF PUBLICATIONS NOT INCLUDED IN THE PROJECTS

Abrashev, R., Dolashka, P., Angelova, M. Temperature pre-treatment modulates oxidative protection of Aspergillus niger cells stressed by paraquat and hydrogen peroxide. Acta Microbiol. Bulg. 2015, 31, 2, 132-140 Abrashev, R., Krumova, E., Kostadinova, N., Miteva-Staleva, J., Spasova, B., Angelova, M. Improvement of superoxide dismutase production by heat shock treatment of Aspergillus niger 26. CR Acad Bulg Sci, 2015, 68, 11, 1379-1386. Belenska L., Gyurkovska, V., Ivanovska, N. How complement activation influences the development of chronic synovitis in a mouse model of rheumatoid arthritis. Scand. J. Rheumatol. 2015, 23, 1-10. Birner, P., Tchorbanov, A., Natchev, S., Tuettenberg, J., Guentchev, M. The chemokine receptor CXCR7 influences prognosis in human glioma in an IDH1 dependent manner. J. Clin. Pathol. 2015, 68, 10, 830-834. Chorukova, E, Simeonov, I. A. Simple mathematical model of the anaerobic digestion of wasted fruits and vegetables in mesophilic conditions. Int. J. Bioautomat. 2015, 19(1), 569-580. Crous, P.W., Wingfield, M.J., Guarro, J., Hernández-Restrepo, M., Sutton, D.A., Acharya, K., Barber, P.A., Boekhout, T., Dimitrov, R.A., Dueñas, M., Dutta, A.K., Gené, J., Gouliamova, D.E., Groenewald, M., Lombard, L., Morozova, O.V., Sarkar, J., Smith M.T., Stchigel A.M., Wiederhold N.P., Alexandrova A.V., Antelmi I., Armengol J. Barnes, I., Cano-Lira, J.F., Castañeda, R.R.F., Contu, M., Courtecuisse, P.R., da Silveira, A.L., Decock, C.A., de Goes, A., Edathodu, J., Ercole, E., Firmino, A.C., Fourie, A., Fournier, J., Furtado, E.L., Geering, A.D., Gershenzon, J., Giraldo, A., Gramaje, D., Hammerbacher, A., He, X.L., Haryadi, D., Khemmuk, W., Kovalenko, A.E., Krawczynski, R., Laich, F., Lechat, C., Lopes, U.P., Madrid, H., Malysheva, E.F., Marín- Felix, Y., Martín, M.P., Mostert, L., Nigro, F., Pereira, O.L., Picillo, B., Pinho, D.B., Popov, E.S., Rodas Peláez, C.A., Rooney-Latham, S., Sandoval-Denis, M., Shivas, R.G., Silva, V., Stoilova-Disheva, M.M., Telleria, M.T., Ullah, C., Unsicker, S.B., van der Merwe, N.A., Vizzini, A., Wagner, H.G., Wong, P.T., Wood, A.R., Groenewald, J.Z. Fungal planet description sheets. Persoonia, 2015, 320-370. Cullen, L., Weiser, R., Olszak, T., Maldonado, R.F., Moreira, A.S., Slachmuylders, L., Brackman, G., Paunova-Krasteva, T.S., Zarnowiec, P., Czerwonka, G., Reilly, J., Drevinek, P., Kaca, W., Melter, O., de Soyza, A., Perry, A., Winstanley, C., Stoitsova, S.R., Lavigne, R., Mahenthiralingam, E., Sá-Correia, I., Coenye, T., Drulis-Kawa, Z., Augustyniak, D., Valvano, M.A., McClean, S. Phenotypic characterization of an international Pseudomonas aeruginosa reference panel: Strains of cystic fibrosis origin show less in vivo virulence than non-CF strains. Microbiology, 2015, doi: 10.1099/mic.0.000155. Delenk, H., Haas, Ch., Gantz, St., Marchev, A., Pavlov, A., Steudler, S. Influence of Salvia officinalis L. hairy roots derived phenolic acids on the growth of Chaetomium globosum and Trichoderma viride. PRO LIGNO, 2015, 11(4), 268-274. Denchev, D., Kabaivanova, L. Resistance of yeast and actinomycete populations at cultivation in a medium with insecticides. Ecol. Eng. Environ. Protect. 2015, 1, 57-64. Dimitrova, L., Kassovski, V., Tzvetkova, I., Michajlova, S., Ivanov, N., Gluhtchev, G., Najdenski, H. Bactericidal effect of electrochemically activated water on the aerobic bacterial population from digestate. Ecol. Eng. Environ. Prot. 2015, 14, 4, 23-32, (in Bulgarian).

Doumanov, J., Mladenova, K., Topouzova-Hristova, T., Stoitsova, S., Petrova, S. Vipoxin and its components affect proliferation and cell death in HepG2 cells. Toxicon 2015, 94, 36- 44. Emilova, N., Denchev, S., Abrashev, R., Cakova, A., Stoycheva, M., Krumova, E. Prognostic significance of extracellular superoxide dismutase in acute coronary syndrome - Gender- specific aspects. CR Acad Bulg Sci. 2015, 68, 4, 529 - 536. Eneva, R., Engibarov, S., Petrova, P., Abrashev, R., Strateva, T., Kolyovska, V., Abrashev, I. High production of neuraminidase by a Vibrio cholerae Non-O1 Strain—the first possible alternative to toxigenic producers. Appl. Biochem. Biotechnol. 2015, 176, 412-427. Fiorentin, L.D., Módenes, A.N., Espinoza-Quiñones, F.R., Trigueros, D.E.G., Kroumov, A.D., Manenti, D.R., Borba, C.E. Biosorption of the reactive blue 5G dye in a fixed bed column packed with orange bagasse: experimental and mathematical modeling, Sep. Sci. Technol. 2015, 50, 2267–2275. Galabov, A.S. Treatment of seasonal flu and basics of prevention for 2015-2016. Medic Art, 2015, 9 (5), 4-8, (in Bulgarian). Galabov, A.S., Gesheva, V., Negoita, T. Viruses in Antarctic habitats: occurrence and ecological importance. Acta Microbiol. Bulg. 2015, 31, 12-20. Galabov, A.S., Uzunov, S., Hadjiathanassova, V., Velichkova, E., Dosseva-Runevska, P., Gegova, G. Anti-influenza virus activity of 1-(4-morpholinomethyl)-tetrahydro-2(1H)- pyrimidinone (Mopyridone). Acta Microbiol. Bulg. 2015, 31, 107-114. Georgieva, A, Tzvetanova, E., Alexandrova, A., Nenkova, G., Mileva, M. Lipid peroxidation in liposomes. In: Proceedings of the sixth workshop on experimental models and methods in biomedical research. IEMPAM, Sofia, 2-14 May 2015, pp 153-160. Georgieva, L, Gadjalova, A., Mihaylova, D., Pavlov, A. Achillea millefolium L. - phytochemical profile and in vitro antioxidant activity. Int. Food Res. J. 2015, 22(4), 1347-1352. Georgieva, L., Ivanov, I., Aneva, I., Denev, P., Georgiev, V., Pavlov, A. protopine production by fumaria cell suspension cultures: effect of light. App. Biochem. Biotechnol. 2015, 176, 287–300. Georgieva, L., Ivanov, I., Marchev, A., Aneva, I., Georgiev, V., Denev, P., Pavlov, A. Initiation and selection of callus cultures from Fumaria rostelata KNAF. As potential producer of isoquinoline alkaloids. Sci. Bull. Series F. Biotechnologies, 2015, 19, 52-57. Gesheva, V., Chausheva, S., Stefanova, N., Mihaylova, N., Doumanova, L., Idakieva, K., Tchorbanov, A. Helix pomatia hemocyanin - a novel bio-adjuvant for viral and bacterial antigens. Int. Immunopharmacol. 2015, 26(1):162-168. Grozdanov, P., Nikolova, N., Galabov, A. Detection of cytomegalovirus (CMV) DNA by PCR in patients with unknown inflammatory eye diseases. J. BioSci. Biotechnol. 2015, SE/ONLINE 43-44. Grozeva, N., Pavlov, D., Petkova, N., Ivanov, I., Denev, P., Pavlov, A., Gerdzhikova. M., Dimanova-Rudolf, M. Characterisation of extracts from Stevia rebaudiana bertoni leaves. Int. J. Pharm. Phytochem. Res. 2015, 7(6), 1236-1243. Gyurkovska, V., Ivanovska, N. Tyrosine kinase inhibitor tyrphostin AG490 reduces liver injury in LPS-induced shock. Eur. J. Pharmacol. 2015, 751, 118-126. Gyurkovska, V., Philipov, S., Kostova, S., Ivanovska, N. Acetylated derivative of glaucine inhibits joint inflammation in collagenase-induced arthritis. Immunopharmacol. Immunotoxicol., 2015, 37, 1, 56-61.

Hristov, A., Christova, N., Kabaivanova, L., Nacheva, L., Stoyneva, I., Petrov, P. Simultaneous biodegradation of phenol and n-hexadecane by immobilized in cryogel biosurfactant producing strain Rhodococcus wratislawiensis BN38. Polish J. Microbiol. 2015, in press. Isachenko, V., Todorov, P., Isachenko, E., Rahimi, G., Tchorbanov, A., Mihaylova, N., Manoylov, I., Mallmann, P., Merzenich, M. Long-time cooling before cryopreservation decreased translocation of phosphatidylserine (Ptd-L-Ser) in human ovarian tissue. PLOS ONE 2015, 10 (6), e0129108. Ivanov, I., Petkova, N., Denev, P., Pavlov, A. Polyphenols content and antioxidant activities in infusion and decoction extracts obtained from Fragaria vesca L. leaves. Sci. Bull. Series F. Biotechnologies 2015, 19, 145-148. Ivanova, J., Kabaivanova, L., Petkov, G. Temperature and irradiance effects on Rhodella reticulata growth and biochemical characteristics Russ. J. Plant Physiol. 2015, 62(5), 690–695. Ivanova, J., Kabaivanova, L., Petrov, P., Yankova, S. Optimization strategies for improved growth, polysaccharide production and storage of the red microalga Rhodella reticulata. Bulg. Chem. Commun. 2015, 47(1), 167-174. Ivanova, A. P, Tsonev, T. D., Peeva, V. N., Maslenkova, L. T., Najdenski, H. M., Tsvetkova, I. V., Babenko, L. M., Shcherbatiuk, M. M., Sheiko, O. A., Kosakivska, I. V. Euhalophyte Eryngium maritimum L the microstructure and functional characteristics. J. Stress Physiol. Biochem. 2015, 11, 3, ISSN: 1997-0838, 52-61 Kabaivanova, L., Chernev, G., Ivanova, J. Construction of inorganic and hybrid biosorbents for heavy metal ions removal. Int. J. Bioautomat. 2015, 19 (4), 473-482. Kabaivanova, L., Ivanova, J., Pehlivanova, V., Nikolova, B. Specific antitumor effect of the combined action of algal heteropolysaccharide and electroporation. Int. J. Bioautomat. 2015, in press. Kalinov, K.N., Ignatova, M.G., Manolova, N.E., Markova, N.D., Karashanova, D.B., Rashkov, I.B. Novel antibacterial electrospun materials based on polyelectrolyte complexes of a quaternized chitosan derivative. RSC Adv. 2015, 5(67), 54517-54526. Lazarkevich, I., Sotirova, A., Avramova, T., Stoitsova, S., Paunova-Krasteva, T., Galabova, D. Antibacterial activity of methyltiosulfonate and its complexes with rhamnolipid and trehalose lipid against Pseudomonas aeruginosa NBIMCC 1390. Res. J. Pharm. Biol. Chem. Sci., 2015, 6(2), 282-290. Litova, K., Manasiev, J., Gerginova, M., Peneva, N., Alexieva, Z. Sequence analyses of genes coding key enzymes responsible for degradation of phenolic compounds by Aspergillus fumigatus strain AL3. J. BioSci. Biotech. 2015, SE/ONLINE: 195-203. Marchev A., Ivanov I., Denev P., Nikolova M., Gochev V., Stoyanova A., Pavlov A., Georgiev V. Acetylcholinesterase inhibitory, antioxidant, and antimicrobial activities of Salvia tomentosa Mill. essential oil. J. BioSci. Biotechnol. 2015, 4(2), 219-229. Markova, N., Slavchev, G., Michailova, L. Presence of mycobacterial L-forms in human blood: Challenge of BCG vaccination. Human. Vaccin. Immunother. 2015, 11(5), 1192-1200. Mihaylova, D., Georgieva, L., Pavlov, A. Antioxidant activity and bioactive compounds of Rosa canina L. herbal preparations. Sci. Bull. Series F. Biotechnologies 2015, 19, 160-165. Mileva, M., Nikolova, I., Nikolova, N., Mukova, L., Georgieva, A., Dobreva, A., Galabov, A.S. Investigation of antioxidant and antiviral properties of Geraniol Acta Microbiol. Bulg. 2015, 31, 48-53.

Miteva-Staleva, J., Krumova, E., Stefanova, T., Angelova, M. Age-related changes in reactive oxygen species production in the filamentous fungus Penicillium rugulosum T35 under cold stress conditions. CR Acad Bulg Sci, 2015, 68, 9, 1123-1128. Mladenova, K., Petrova, S., Moskova-Doumanova, V., Topouzova-Hristova, T., Stoitsova, S., Tabashka, I., Chakarova, C., Lalchev, Z., Doumanov, J. Transepithelial resistance in human bestrophin-1 stably transfected Madin-Darby canine kidney cells. Biotechnol. Biotechnol. Equip. 2015, 29, 101-104. Módenes, A.N., Espinoza-Quiñones, F.R., Geraldi, C.A.Q., Manenti, D.R., Trigueros, D.E.G., de Oliveira, A.P., Borba, C.E., Kroumov, A.D. Assessment of the banana pseudostem as low-cost biosorbent for removal of the reactive blue 5G dye. Environ. Technol. 2015, 36: (22), 2892–2902. Módenes, A.N., Scheufele, F.B., Barbosa, J.C., Colombo, A., Trigueros, D.E.G., Espinoza- Quiñones, F.R., Hinterholz, C.L., Kroumov, A.D. Assessment of kinetic, equilibrium and thermodynamic of black Krom KJR dye adsorption onto aquatic macrophyte Pistia stratiote. Environ. Eng. Manag. J. 2015, in press. Módenes, A.N., Scheufele, F.B., Espinoza-Quiñones, F.R., de Souza, P.S.C., Cripa, C.R.B., dos Santos, J., Steffen, V., Kroumov, A.D. Adsorption of direct of yellow ARLE dye by activated carbon of shell of coconut palm: Kinetics, equilibrium and mechanism study, Int. J. Bioautomat. 2015, 19(2), 187-206. Nemska, V., Georgieva, N., Danova, S. Evaluation of technological parameters of newly isolated lactobacilli from traditional dairy products. Scientific Conference “Tradition and Innovation in the chemical, food and biotechnology” University of Ruse, Branch Razgrad, 16-17.10.2015, pp. 9-14. Nemska, V., Georgieva, N., Danova, S. Initial characterization and antibiotic susceptibility of lactic acid bacteria isolated from traditional dairy products. 62nd Scientific Conference „ Food Science, Engineering and Technology 2015”, University of Food Technologies 23.10.2015, pp. 510-513. Paunova-Krasteva, T., Ivanova, R., De Castro, C., Molinaro, A., Stoitsova, S. Promotion of the synthesis of a concanavalin A-reactive polysaccharide upon growth of Escherichia coli O157:H(-) on solid medium at 37oC. Acta Microbiol. Bulg. 2015, 31(2), 122-126. Pavlova, V., Paunova-Krasteva, T., Stoitsova, S., Nikolova, E. Distribution patterns of carbohydrates in murine glycocalyx. Biotechnol. Biotechnol. Equip. 2015, 29, 357-362. Pavlova-Hristova, I., Danova, S., Naidenski, H., Tropcheva, R., Milanova, A. Effect of probiotics on Enrofloxacin disposition in gastrointestinal tract of poultry. J. Vet. Pharmacol. Therapeut. 2015, 38, 6, 549-555. Petkova, N., Ivanov, I., Topchieva, S., Denev, P., Pavlov, A. Biologically active substances and in vitro antioxidant activity of different extracts from dandelion (Taraxacum officinale) roots. Sci. Bull. Series F. Biotechnologies 205, 19, 190-197. Petkova, N., Vrancheva, R., Mihaylova, D., Ivanov, I., Pavlov, A., Denev, P. Antioxidant activity and fructan content in root extracts from elecampane (Inula helenium L.). J. BioSci. Biotechnol. 2015, 4(1), 101-107. Petrov, N. M., Galabov, A.S. RNAi - strategy to control viral infections in eukaryotic organisms. Acta Microbiol. Bulg. 2015, 31, 21-31. Radchenkova, N., Panchev, I., Vassilev, S., Kuncheva, M., Dobreva, S., Kambourova, M. Continuous cultivation of a thermophilic bacterium Aeribacillus pallidus 418 for production of an exopolysaccharide applicable in cosmetic creams. J. Appl. Microbiol. 2015, 119(5), 1301-1309.

Rusinova-Videva, S., Radchenkova, N., Dobrev, G., Pavlova, K. Purification of arabinomannan synthesized by Cryptococcus laurentii AL100. Acta Microbiol. Bulg. 2015, 31, 2, 141- 144. Stoyancheva, G., Chorukova, E. Lactobacillus gasseri G7- a bacteriocinogenic strain isolated from vaginal sample. Annuaire de l’Université de Sofia “St. KlimentOhridski” 2015, 100, 4, 154-163. Stoykov, Y., Pavlov, A., Krastanov, A. Chitinase biotechnology: Production, purification and application. Eng. Life Sci. 2015, 15(1), 30–38. Teneva-Angelova, T., Beshkova, D. Genus Salvia – ecosystem for isolation of lactic acid bacteria. J. Microbiol. Biotechnol. Food Sci. 2015, 5, 2, 103-108. Teneva-Angelova, T., Beshkova, D. Resistance profile of plant-derived lactic acid bacteria against herb extracts. Sci. Bull., Series F, Biotechnologies, 2015, XIX, 109-116. Teneva-Angelova, Ts., Beshkova, D. Non-traditional sources for isolation of lactic acid bacteria. Ann. Microbiol. 2015, in press. Tropcheva, R., Lesev, N., Danova, S., Stoitsova, S., Kaloyanova, S. Novel cyanine and homodimeric styryl dyes as fluorescent probes for assessment of lactic acid bacteria cell viability. J. Photochem. Photobiol. B: Biol. 2015, 143, 120-129. Tropcheva, R., Lesev, N., Danova, S., Stoitsova, S., Kaloyanova, S. Novel cyanine dyes and homodimeric styryl dyes as fluorescent probes for assessment of lactic acid bacteria cell viability. J. Photochem. Photobiol. B: Biology 2015, 143, 120–129. Yakovleva, M., Tacheva, T., Mihaylova, S., Tropcheva, R., Trifonova, K., Tolekova, A., Danova, S., Valyakova, T. Influence of Lactobacillus brevis 15 and Lactobacillus plantarum13 on blood glucose and body wheigth in rats after high-fructose diet. Benef. Microbes 2015, 6, 4, 505-512. Yildiz, S. Y., Radchenkova, N., Arga, K. Y., Kambourova, M., Oner, E. T. Genomic analysis of Brevibacillus thermoruber 423 reveals its biotechnological and industrial potential. Appl. Microbiol. Biotechnol. 2015, 99(5), 2277-2289.

Textbooks: Ganova, P., Gyurkovska, V., Ivanovska, N. Tumour necrosis factor-related apoptosis inducing ligand (TRAIL) as an arthritic marker in a model of osteoarthritis. Sixth Workshop on Experimental models and Methods in Biomedcal Research, 2015, ISSN:1314-9091, 21 – 25. Gyurkovska, V., Dimitrova P., Ivanovska, N. Plant remedies: How they can overcome Candida infections. Natural Products: Research Reviews, 3, Daya Publishing House®, 2015, 327 – 351, ISBN:9789351246855. Kambourova, M. Recent advances in extremophilic amylases. In: Extremophilic Bioprocessing of Lignocellulosic Feedstocks to Biofuels, Value-Added Products, and Usable Power (Sani, R.K., Ed.). Springer International Publishing AG, Gewerbestrasse 11, 6330 Cham, Switzerland. 2015, in press. Kambourova, M., Radchenkova, N., Tomova, I., Bojadjieva, I. Thermophiles as a promising source of exopolysaccharides with interesting properties. In: Biotechnology of Extremophiles: Advances and Challenges (Pabulo, R., Ed.) 2015, in press. Kambourova, М., Toksoy Oner, E., Poli, A. Exopolysaccharides from prokaryotic microorganisms - promising sources for white biotechnology processes. In: Industrial Biorefineries and White Biotechnology (Pandey, A., Höfer, R., Taherzadeh, M.,

Nampoothiri, K. M., Larroche, C., Eds) Elsevier, Amsterdam, Netherlands, 2015, pp. 523-554. Konstantinov, S., Zaharieva, M.M., Argirova, R., Najdenski H. Acute inflammatory diseases of the upper respiratory tract – types and therapeutic modalities. Acute bronchitis. In: Pharmacotherapy, S. Konstantinov (ed). Softrade, Sofia, 2015, pp. 213-219, (in Bulgarian). Konstantinov, S., Zaharieva, M.M., Momekov, G., Najdenski, H. Pneumonia - etiology, pathogenesis, types and chemotherapy. In: Pharmacotherapy, S. Konstantinov (ed). Softrade, Sofia, 2015, pp. 220-238, (in Bulgarian). Mladenov, K., Najdenski, H. Studies of environmental foci of tularemia in the West of Sofia and North-West Bulgaria. In: Tularemia - rabbit fever, Impera Print, 2015, ISBN 978- 619-90471-0-1, (in Bulgarian). Mladenov, K., Najdenski, H. Tularemia in humans. In: Tularemia - rabbit fever, Impera Print, 2015, ISBN 978-619-90471-0-1, (in Bulgarian). Mladenov, K., Mekushanov K., Najdenski, H. Tularemia in Bulgaria. In: Tularemia - rabbit fever, Impera Print, 2015, ISBN 978-619-90471-0-1, (in Bulgarian). Najdenski, H. Tularemia in wild and domestic animals. In: Tularemia - rabbit fever, Impera Print, 2015, ISBN 978-619-90471-0-1, (in Bulgarian). Zaharieva, M.M., Konstantinov, S., Momekov, G., Argirova, R., Najdenski, H. Chemotherapy resistance - types and mechanisms of resistance, classical and modern therapeutic modalities. In: Pharmacotherapy, S. Konstantinov (ed). Softrade, Sofia, 2015, pp. 60-88, (in Bulgarian). Zaharieva, M.M., Najdenski, H. Infectious inflammatory syndrom - causative agents and clinical manifestation depending on the localication of the infection and the choice of chemotherapy. In: Pharmacotherapy, S. Konstantinov (ed). Softrade, Sofia, 2015, pp. 112-140, (in Bulgarian).

EDUCATION ACTIVITY Lectures and Practical Exercise

Acad. A.S. Galabov, DSc Course in Medical Virology, Kliment Ohridski University of Sofia, Faculty of Biology Course in Antiviral Therapy, Kliment Ohridski University of Sofia, Faculty of Biology Course in General Virology, New Bulgarian University

Corr. Member H. Najdenski, DSc Course in Infectious Diseases, University of Forestry, Faculty of Veterinary Medicine, Sofia Practical exercises in Infectious Diseases, University of Forestry, Faculty of Veterinary Medicine, Sofia Course in Molecular Methods in Microbiology, New Bulgarian University Practical exercises in Molecular Methods in Microbiology, New Bulgarian University Course in Nutrition and Dietetic, New Bulgarian University Practical exercises in Nutrition and Dietetic, New Bulgarian University

Corr. Member A. Pavlov, DSc Course in Food Chemistry (for BS students), University of Food Technologies, Plovdiv Practical exercises in Food Chemistry (for BS students), University of Food Technologies, Plovdiv Course in Bioactive Substances from Cell Cultures (for MS students), University of Food Technologies, Plovdiv Practical exercises in Bioactive Substances from Cell Cultures (for MS students), University of Food Technologies, Plovdiv Course in Biotechnological Productions based on Plant and Cell Cultures (for BS students), Agricultural University, Plovdiv Practical exercises in Biotechnological Productions based on Plant and Cell Cultures (for BS students), Agricultural University, Plovdiv Course in Modern Technologies and Methods for Analysis of Bioactive Substances Chemistry (for BS students), University of Food Technologies, Plovdiv Practical exercises in Modern Technologies and Methods for Analysis of Bioactive Substances Chemistry (for BS students), University of Food Technologies, Plovdiv

Prof. M. Angelova, DSc Course in Oxidative Stress (for BS students), New Bulgarian University Course in Mycology (for BS students), New Bulgarian University Course in Mycology (for BS students), Plovdiv University Paisii Hilendarski.

Assoc. Prof. L. Doumanova, PhD Course in Immunochemistry (for MS students), St Kliment Ohridski University of Sofia, Faculty of Chemistry Practical exercises in Immunochemistry (for MS students), St Kliment Ohridski University of Sofia, Faculty of Chemistry

Assoc. Prof. S. Danova, PhD Course in The Probiotics (for MS students), University of Chemical Technology and Metalurgy, Sofia, Department of Industrial Biotechnology

Assoc. Prof. S. Stoitsova, PhD Course in Methods of Electron Microscopy, Histochemistry and Immunocytochemistry in Cell Biology, St. Kliment Ohridski University of Sofia, Faculty of Biology Practical exercises in Methods of Electron Microscopy, Histochemistry and Immunocytochemistry in Cell Biology, St. Kliment Ohridski University of Sofia, Faculty of Biology

Assoc. Prof. Andrey Tchorbanov, PhD Course in Immunology (for BS students), New Bulgarian University Practical exercises in Immunology (for BS students), New Bulgarian University Course in Animal Models for Immunology of Reproduction, Project for young investigators and PhD students, №BG051PO001-3.3.06-0059, Institute IBIR, BAS

Assoc. Prof. D. Beshkova, PhD Course in Microbial Safety of Animal Products, (for MS students), University of Food Technologies, Plovdiv Course in Microbiology of Fermented Functional Foods (for MS students), University of Food Technologies, Plovdiv

Assoc. Prof. E. Krumova, PhD Practical exercises in Oxidative Stress (for BS students), New Bulgarian University Practical exercises in Mycology (for BS students), New Bulgarian University Practical exercises in Cytology, Hyistology and Embryology St. Kliment Ohridski University of Sofia, Faculty of Biology

Assoc. Prof. M. Mileva, PhD Course in Pharmacologyo of Eye Diseases (for MS students), St. Kliment Ohridski University of Sofia, Faculty of Physics Practical exercises in Pharmacology of Eye Diseases (for MS students), St. Kliment Ohridski University of Sofia, Faculty of Physic Course in Analytical Chemistry and Analysis of Medicinal Products, Medical College "Yordanka Filaretova", Sofia Practical exercises in Analytical Chemistry and Analysis of Medicinal Product, Medical College "Yordanka Filaretova", Sofia

Assis. Prof. V. Nicolova, PhD Practical exercises in Virology, New Bulgarian University

Assist. Prof. S. Rusinova-Videva, PhD Practical exercise in Micology (for BS students), Plovdiv University Paisii Hilendarski, Plovdiv Practical exercise in Hydrobiology (for BS students), Plovdiv University Paisii Hilendarski, Plovdiv Practical exercise in Hydrobiology (for BS students), Plovdiv University Paisii Hilendarski, Branch Smolyan

Assis. Prof. N. Kostadinova, PhD Practical exercises in Cytology and Cell Biology, St. Kliment Ohridski University of Sofia, Faculty of Biology

Assist. Prof. A. Marchev, PhD Practical exercises in Biotechnological Productions Based on Plant and Cell Cultures (for BS students), Agricultural University, Plovdiv

Assist. Prof. R. Tropcheva, PhD Practical exercises on Secondary Metabolites Biotechnology, St Kliment Ohridski University of Sofia, Faculty of Biology

Assis. Prof. A. Kroumov, PhD Course in Photobioreactors-modeling, optimization, design and scale up, West Parana State University, Toledo, Parana, Brazil

Defended DSc Thesis

Danova S. Biodiversity and probiotic potential of lactic acid bacteria from different ecological niches

Defended PhD Thesis

Gesheva V. Modulation of immune response by hemocyanins from Rapana thomasiana Advisor: Assoc. Prof. A. Tcorbanov, PhD and Assoc. Prof. K. Idakieva, PhD Hubenov, V. Studies of anaerobic digestion of organic wastes at mesophilic and thermophilic conditions”, Advisors: Assoc. Prof. I. Simeonov, PhD and Assoc. Prof. D. Denchev, PhD Kerekov N. Humanized experimental models of autoimmune and allergic diseases. Advisor: Assoc. Prof. A. Tcorbanov, PhD Milanova V. Inflammatory and destructive potential of neutrophiles in arthritis. Advisor: Assoc. Prof. P. Dimitrova, PhD Pashova S. Applied investigations of B cell populations. Advisor: Assoc. Prof. A. Pashov, PhD

PhD students

Belenska L. Role of complement in the processes of joint destruction in experimental models of arthritis. Advisor: Prof. N. Ivanovska, DSc Dimitrova L. New antimicrobial agents with therapeutic potential Advisors: Corr. Member H. Najdenski, DSC and Corr. Member V. Bankova, DSc Ganchev I. Mixed biofilms. Advisor: Assoc. Prof. S. Stoitsova, PhD

Ganova P. Influence of tyrosine kinase inhibitors on the processes of osteoclastogenesis in experimental arthritis. Advisor: Prof. N. Ivanovska, DSc Gotova I. Study of probiotic characteristics of lactic acid bacteria and selection of strains for products with health benefits Advisors: Corr. Member H. Najdenski, DSC and Assoc. Prof. J. Dimitrov, PhD Hadjieva M. Induced polyspecificity of antibodies. Advisor: Prof. T. Vassilev, MD, DSc Hinterholz C. L. Cultivation of algae in a new photo-bioreactor for the sequestration of carbon dioxide from waste gases of various types and synthesis of high value products. West Parana State University, Toledo PR, Brazil. Advisor: Assis. Prof. A. Kroumov, PhD Ivanova I. Antigen targeting by genetically engineered chimeric molecules Advisor: Assoc. Prof. A. Tchorbanov, PhD Litova K. Microbial biodegradation of industrial pollutants of the environment Advisor: Assoc. Prof. Z. Alexieva, PhD Manoilov I. Specific therapy of autoimmune diabetes in mouse and humanized models. Advisor: Assoc. Prof. A. Tchorbanov, PhD Miteva-Staleva J. Cold stress and aging in Antarctic fungi Advisor: Prof. M. Angelova, DSc Nemska V. Biotechnological and functional characteristics of probiotic lactic acid bacteria Advisor: Assoc. Prof. Svetla Danova, DSc Stoyanov A. Genetic and molecular studies on methylotrophic yeast Hansenula polymorpha Advisor: Assoc. Prof. K. Lahtchev, PhD Stoykov Y. Production and haracterization of microbial chitinases Advisors: Prof. A. Pavlov, DSc and Prof. A. Krastanov, DSc Teneva Ts. Biodiversity of lactic acid bacteria in Bulgarian medical plants and solution for using their biological potential Advisor: Assoc. Prof. D. Beshkova, PhD Velikova P. Polysaccharide-modifying lactic acid bacteria with application in the food industry Advisor: Assoc. Prof. P. Petrova, PhD Vrancheva R. Bioacive substanses from Bulgarian Fumaria spp. Advisor: Prof. A. Pavlov, DSc Zahmanov G. Pharmaceutically important metabolites from from Sambucus ebulus and its in vitro cultures Advisor: Assoc. Prof. M. Georgiev, PhD

Defended MS Diploma Thesis

Adamov A. Biosorption potential of biomass of Antarctic strain Cryptococcus laurentii AL65. Advisor: Assist. Prof. S. Rusinova-Videva, PhD Andrejnski N. Cell response of two strains Aspergillus fumigatus, isolated from heavy metal polluted regions against metal-induced toxicity Advisor: Assoc. Prof. E. Krumova, PhD

Bratanova D. Study on Lactobacillus microbiota of breast milk Advisor: Assoc. Prof. Svetla Danova, DSc Boteva N. Studies at newly isolated Lactobacilli from fermented dairy products Advisor: Assoc. Prof. Svetla Danova, DSc

Defended BS Diploma Thesis Delcheva N. Suppression of auto-reactive B and T lymphocytes from diabetis patients by protein chimeric molecules. Advisor: Assoc. Prof. A. Tchorbanov, PhD Dyankova D. Antioxidant enzyme defence of filamentous fungi isolated from heavy metal polluted soils Advisor: Assoc. Prof. E. Krumova, PhD Kolev K. Selective elimination of specific B-lymphocytes by protein-engineered chimeric molecules. Advisor: Assoc. Prof. A. Tchorbanov, PhD Petkova N. Production and characterization of the antioxidants enzyme superoxide dismutase from the strain Aspergillus fumigatus 32, resistant to heavy metals Advisor: Assist. Prof. R. Abrashev, PhD Slavov S. Functional elimination of auto-reactive T cells in murine MRL/lpr model of lupus by therapy with antibody against Annexin A1. Advisor: Assoc. Prof. A. Tchorbanov, PhD

Postdoc practice Semcheddine S. Maitre de conference Advisor: Assoc. Prof. Ivan Simeonov, PhD

National Student Practice financed by Operational Programme “Human Resources Development”, co-financed jointly by the European Social Fund of the EU Bradyanova S. Experimental immunology Assoc. Prof. A. Tchorbanov, PhD Chepinski P. Experimental immunology Advisor: Assoc. Prof. A. Tchorbanov, PhD Delcheva N. Experimental immunology Advisor: Assoc. Prof. A. Tchorbanov, PhD Hlebarska D. Experimental immunology Advisor: Assoc. Prof. A. Tchorbanov, PhD Slavov S., Experimental immunology Advisor: Assoc. Prof. A. Tchorbanov, PhD Valentinov K., Experimental immunology Advisor: Assoc. Prof. A. Tchorbanov, PhD Boneva G., Experimental immunology Advisor: Assoc. Prof. A. Tchorbanov, PhD Kostadinova V., Experimental immunology Advisor: Assoc. Prof. A. Tchorbanov, PhD

SCIENTIFIC EVENTS

CELEBRATION DAY OF THE STEPHAN ANGELOFF INSTITUTE OF MICROBIOLOGY The Day of Institute, March 14, already turned into tradition, was celebrated with a ceremonial meeting. The meeting was opened with exciting speech delivered by the Director - Prof. H. Najdenski, Corresponding Member of BAS. Special guest and invited speaker in 2015 was Prof. Lydmila Boyanova, MD, DSc, from Medical University of Sofia that gave a very intriguing lecture entitled “Helicobacter pylori – the carcinogenic bacteria”. Acad. Angel S. Galabov, delivered the traditional Annual Prize for the best scientific publication by young microbiologist (under 35 year of age) to Tsvetelina Paunova-Krusteva, PhD (The Stephan Angeloff Institute of Microbiology, Department of General Microbiology).

ACTIVITIES OF FOUNDATION “ACAD. PROF. DR. STEPHAN ANGELOFF” The annual awards for the best scientific publication by young microbiologist (under 35 year of age) were been given to Tsvetelina Paunova-Krusteva, PhD (The Stephan Angeloff Institute of Microbiology, Department of General Microbiology) for the following paper: Paunova-Krasteva Ts., Pavlova V., DeCastro C., Ivanova R., Molinaro A., Nikolova E., Stoitsova S. (2014). Cyclic enterobacterial common antigens from Escherichia coli O157 as microbe-associated molecular patterns. Can. J. Microbiol., 60, 173-176.

SCIENTIFIC SYMPOSIUM „BENEFICIAL AND PATHOGENIC MICROBES FOR HEALTHIER LIFE AND SAFETY FOODS”. Chairpersons: Assoc. Prof. SVETLA DANOVA, DSc and Corr. Member HRISTO NAJDENSKI, DSc The Stephan Angeloff Institute of Microbiology (Bulgarian Academy of Sciences) in cooperation with the United Scientific Group (USG) hosted the scientific symposium Beneficial and pathogenic microbes for healthier life and safety foods on April 8-9, 2015 at Institute of Microbiology, Acad. G. Bontchev, str. 26, 1113 Sofia, Bulgaria. The two days scientific symposium, was a pre-conference event of the Probiotic summit 2015 San Francisco, USA (http://www.unitedscientificgroup.com/conferences/probiotics-summit/ The symposium explored critical points in application of beneficial microbes – probiotics and the risk of food-borne pathogens:  for safety and functional food;  new approaches for prevention the microbial risks in food chain from pathogens;  new trends in protection of human and animals health;  prophylaxis and therapy of food disorders for wellbeing and health. The symposium provided an opportunity to the participating scientists and young researchers to discuss various issues pertaining to their research career. It gived an understanding of the beneficial and pathogenic microbes, functional and safety foods for long and healthier life. The symposium assembled scientists from different Universities/Institutions (Bulgarian Academy of Sciences, Sofia University “St. Kliment Ohridski”, Plovdiv University “Paisij Hilendarski”, Bulgarian Food Safety Agency, Center for Risk Assessment, etc.) that have developed innovative researches and analyses to these problems and discussions by education leaders about applications and solutions for other activities that could relieve critical situation and improve the know-how and efficiency of the coming generation. Site of the event: www.microbio.bas.bg/conferences/probiotics

NATIONAL CONFERENCE WITH INTERNATIONAL PARTICIPATION “ECOLOGICAL ENGINEERING AND ENVIRONMENT PROTECTION” Fourth National Conference with International Participation and Youth Scientific Session „Ecological Engineering and Environmental Protection” EEEP was held on 03-06 June 2015, at Burgas, Bulgaria.

INTERNATIONAL IMMUNOLOGY SCHOOL, Organizer: Assoc. Prof. ANDREJ TCHORBANOV, PhD - President of Bulgarian Society for Immunology (BuSI) 2nd Black sea International Immunology school (BSIIS2015) was held on 15-16 October 2015 in Hotel “Legends” - Sofia, Bulgaria. The scool was funded by EFIS.

AWARDS FOR 2015 Award “Pitagor” Assoc. Prof. Milen Georgiev, PhD (Department of Applied Microbiology) got the Winner in the "Pythagoras" award for an established scientist in the technical sciences. Awards of the Union of Scientists in Bulgaria - 2015 Assoc. Prof. Petya Dimitrova, PhD (Department of Immunology) received a Diploma of the Bulgarian Union of Scientists for Excellence thesis of her PhD student Victoria Milanova. Assist. Prof. Kalina Nikolova-Ganeva, PhD (Department of Immunology) was awarded a price of the Bulgarian Union of Scientists for 2015 year in the field of Biology and Medicine. Assist. Prof. Tsvetelina Paunova-Krusteva, PhD (Department of General Microbiology) was awarded a price of the Bulgarian Union of Scientists for excellence in PhD thesis for 2015. Award of Evrika Foundation Assoc. Prof. Petya Dimitrova, PhD (Department of Immunology) was awared a price of Evrika Foundation for for Excellence thesis of her PhD student Victoria Milanova Awards of Congress and Conferences Assoc. Prof. Petya Dimitrova, PhD (Department of Immunology) received the TEWA award for the best oral presentation on 2nd ICNPU, Plovdiv, Bulgaria. Assist. Prof. Maya Zaharieva, PhD (Department of Infectious Microbiology) received the TEWA award for the best poster on 2nd ICNPU, Plovdiv, Bulgaria.

Staff of the Stephan Angeloff Institute of Microbiology

SCIENTIFIC DIVISION

DEPARTMENT OF GENERAL Stephan Engibarov, MS, Assist. Professor MICROBIOLOGY Irina Lazarkevich, MS, Assist. Professor

Head of Department: Zlatka Aleksieva, DEPARTMENT OF APPLIED PhD, Assoc. Professor MICROBIOLOGY

Section of Morphology of Microorganisms Head of Department: Atanas Pavlov, DSc, and Electron Microscopy Professor

Head: Stoyanka Stoitsova, PhD, Assoc. Laboratory of Microbial Biosynthesis and Professor Ecology Tsvetelina Paunova, PhD, Assist. Professor Head: Kolishka Tcekova, PhD, Assoc. Ivan Chavdarov, MS, Electronic Engineer Professor Dayana Borisova, MS, Microbiologist Adriana Gousterova, PhD, Assoc. Professor Nina Tadjer, BS, Laboratory Assistant Lyudmila Kabaivanova, PhD, Assoc. Professor Lidana Panova, BS, Laboratory Assistant Aleksander Krumov, PhD, Assist. Professor Ivo Ganchev, PhD student Dessislava Todorova, PhD, Assist. Professor Section of Microbial Genetics Ignat Dimov, Assist. Professor Lilyana Nacheva, Assist. Professor Head: Zlatka Aleksieva, PhD, Assoc. Philip Philipov, Researcher Professor Rositca Petkova, Researcher Svetla Danova, PhD, Assoc. Professor Anna Brachkova, PhD student Penka Petrova, PhD, Assoc. Professor Maria Gerginova, PhD, Assist. Professor Laboratory of Extremophilic Bacteria Margarita Stoilova, PhD, Assist. Professor Head: Margarita Kambourova, DSc, Assoc. Nadejda Peneva, MS, Assist. Professor Professor Jordan Manasiev, PhD, Assist. Professor Ivanka Bojadzhieva, PhD, Assist. Professor Dimitrinka Lyutskanova, PhD, Assist. Miroslava Atanasova PhD, Assist. Professor Professor Iva Tomova, PhD, Assist. Professor Galina Stoyancheva, PhD, Assist. Professor Nadja Kirilova, PhD, Assist. Professor Dillnora Gouliamova, PhD, Researcher Nikolina Atanasova, MS, Assist. Professor Rositza Tropcheva, PhD, Assist. Professor

Vesela Peltekova, Technician Anton Stojanov, PhD student Laboratory of Applied Biotechnologies Section of Microbial Biochemistry Head: Atanas Pavlov, DSc, Professor Head: Evgenia Vasileva-Tonkova, PhD, Milen Georgiev, PhD, Assoc. Professor Assoc. Professor Kostanca Pavlova, PhD, Assoc. Professor Blaga Angelova, PhD, Assoc. Professor Dora Beshkova, PhD, Assoc. Professor Nelly Christova, PhD, Assist. Professor Ginka Frengova, PhD, Assoc. Professor Rumyana Eneva, PhD, Assist. Professor Vasil Georgiev, PhD, Assist. Professor Dilyana Simeonova, PhD Assist. Professor Snejana Videva, PhD, Assist. Professor Anna Sotirova, MS, Assist. Professor

Andrei Marchev, PhD Assist. Professor Tchavdar Tankov, Laboratory Assistent Katerina Georgieva, MS, Microbiologist Tatyana Gocheva, Technologist DEPARTMENT OF VIROLOGY Elka Genova, Technologist Svetla Stojkova, Technologist Head of Department: Ivanka Nikolova, Vasilka Ushterova, Laboratory Assistant PhD, Assist. Professor

Tzvetanka Teneva, PhD student Academician Angel S. Galabov, MD, DSc, Yuri Stoikov, PhD student Professor Georgi Zahmanov, PhD student Laboratory of Viral Proteins Research group of Mathematical modelling Head: Lyubka Doumanova, PhD, Assoc. and Computer Methods Professor Head: Ivan Simeonov, PhD, Assoc. Milka Mileva, PhD, Assoc. Professor Professor Yurii Abashev, PhD, Assist. Professor Elena Chorukova, PhD, Assist. Professor Adriana Dimitrova, PhD student Venelin Hubenov, Assist. Professor Snezhanka Mihaylova, MS, Technologist Laboratory of Experimental Georgi Valevski, Dipl. eng. Chemotherapy of Enteroviral Infections Head: Ivanka Nikolova, PhD, Assist. DEPARTMENT OF INFECTIOUS Professor MICROBIOLOGY Nadya Nikolova, MS, Researcher Snejana Andreeva, Laboratory Assistant Head of Department: Hristo Najdenski, DVM, DSc, Professor, Corr. Member of Laboratory of Experimental Chemotherapy BAS of Influenza Laboratory of Zoonozes and Bacterial Head: Lora Simeonova, PhD, Assist. Virulence Professor Head: Hristo Najdenski, DVM, DSc, Galina Gegova, MS, Researcher Professor, Corr. Member of BAS Kirilka Todorova, BS, Technician Maya Zaharieva, PhD, Assist. Рrofessor Violeta Valcheva, PhD, Assist. Рrofessor Laboratory of Oncolytic Viruses Zvezdimira Tcvetanova, PhD, Assist. Рrofessor Head: Assya Angelova, PhD, Assist. Magdalena Bonovska, PhD, Researcher Professor Trayana Draganova, MS, Researcher Zahari Raykov, MD, PhD, Assist. Professor Dimitar Dimitrov, Assist. Рrofessor Neli Vilhelmova, PhD, Assist. Professor Vanja Slaveva, Researcher Adelina Stoyanova, MS, Assist. Professor Maya Gatzovska, MS Luchia Mukova, MS, Researcher Laboratory of Genetics and Resistance of Mycobacteria DEPARTMENT OF IMMUNOLOGY Head: Nadya Markova, MD, PhD, Assoc. Professor Head of Department: Nina Ivanovska, DSc, Georgi Slavchev, MS, PhD student Professor Albena Cherneva, Researcher Laboratory of Experimental Immunotherapy Laboratory of Antimicrobial Agents Head: Anastas Pashov, PhD, Assoc. Head: Vesselin Kussovski, PhD, Assoc. Professor Professor Tchavdar Vassilev, MD, DSc, Professor Iva Tzvetkova, MS, Researcher Petya Dimitrova, PhD, Assoc. Professor 51

Iglika Djoumerska-Alexieva, MD, PhD, Petya Ganova, PhD student Assist. Professor Maya Hadjieva, PhD student SECTION OF MYCOLOGY Head: Ekaterina Krumova, PhD, Assoc. Laboratory of Experimental Immunology Professor Head: Andrey Tchorbanov, PhD, Assoc. Maria Angelova, DSc, Professor Professor Radoslav Abrashev, PhD, Assist. Professor Nikolina Mihaylova, PhD, Assist. Professor Nedelina Kostadinova, PhD, Assist. Kalina Nikolova-Ganeva, PhD, Assist. Professor Professor Jeni Miteva-Staleva, MS, Assist. Professor Ilian Manoilov, PhD, Assist. Professor Vladislava Dishliiska, MS, Assist. Professor Nikola Kerekov, PhD, Assist. Professor Borjana Spasova, MS, Researchеr Viktoriya Milanova, PhD student Emilia Eftova, Laboratory Assistant Laboratory of Infectious Immunology and Inflammation LABORATORY CENTER PASTEUR Head: Nina Ivanovska, DSc, Professor Head: Peter Grozdanov, PhD, Assist. Tsvetanka Stefanova, PhD, Assist. Professor Profеssor Valeriya Gyurkovska, PhD, Assist. Professor Anna Terziyska, PhD, Assist. Professor Petya Stoyanova, DVM, Researcher Iva Ivanova, PhD student

ADMINISTRATIVE AND TECHNICAL DIVISION Dima Kuzarova, MS Secretariat Krasimira Beshkova, Technician Petya Nikolova, Coordinator Nadka Panova, Human Resources Fermentation Laboratory Ivailo Georgiev, MS, IT Serviceр eng Spasen Vassilev, Assist. Professor Krasya Nenova, Technical secretary Vesselina Pankova, MS, Researcher eng. Ivan Georgiev, Secretary Galina Nikolova, MS Nora Toupareеva, Courrier Animal House Facility Accountancy Petya Stoyanova, DVM, Researcher Antoaneta Tcareva, Chief accountant Eleni Axioti Dimitrova Stefka Yonkova, Accountant Ivanka Zachova Snezhanka Daskalova, Accountant-cachier Violeta Manolova, Accountant Technical Service Evgenia Minkova, Asssitant director, eng. Еlectro-Mechanical Workshops Public Relations Dimiter Brajnov Violeta Valcheva, PhD, Assist. Professor Plamen Stefkov Lora Simeonova, PhD, Assist. Professor Tzvetanka Begova Library Todorka Kehajova, Librarian Anna Vaneva, Researcher

BIOLOGICAL SERVICES

Laboratory Cultural Media Nataliya Georgieva, MS, Researcher 52

SCIENTIFIC BOARD

Chairman: Zlatka Alexieva Associate Professor, PhD Vice-Chairman: Tchavdar Vassilev Professor, DSc Secretary: Penka Petrova Associate Professor, PhD

Angel S. Galabov Professor, DSc, Academician of BAS Hristo Najdenski Professor, DSc Todor Kantardjiev Professor, DSc Maria Angelova Professor, DSc Nina Ivanovska Professor, DSc Atanas Pavlov Professor, DSc Vera Maximova Associate Professor, PhD Lyubka Doumanova Associate Professor, PhD Blaga Angelova Associate Professor, PhD Danka Galabova Associate Professor, PhD Margarita Kamburova Associate Professor, PhD Stoyanka Stoitsova Associate Professor, PhD Lubomira Nikolaeva-Glomb Associate Professor, PhD Veselin Kasovski Associate Professor, PhD Nadya Markova Associate Professor, PhD Svetla Danova Associate Professor, PhD Andrey Chorbanov Associate Professor, PhD Petja Dimitrova Associate Professor, PhD

INTERNATIONAL SCIENTIFIC BOARD

Fabian Wild Professor, Expert in the Centre of WHO, Lion, France Jeffery Almond Professor, Head of the School of Animal and Microbial Sciences at the University of Reading, UK and Vice-President, Discovery Research and External Research and Development at Sanofi Pasteur Anna Erdei Professor, Head of Department of Immunology, University of L. Eotvos, Budapest, Hungary Fergus G. Priest Professor, Head of Life Sciences, International Centre for Brewing and Distilling, School of Life Sciences Heriot-Watt University Edinburgh, UK Erik DeClercq Professor, Director of Rega Institute for Medical Research at the Catholic University of Leuven, Belgium Dietmar Fuchs Professor, University of Innsbruck, Austria Tomas Bley Professor, Chair of Bioprocess Engineering, Institute of Food Technology and Bioprocess Engineering

NATIONAL SCIENTIFIC SEMINARS*

General Microbiology Chairman: Assoc. Prof. S. Danova, DSc Vice Chairman: Assoc. Prof. S. Stoitsova, PhD Secretary: Assist. Prof. Ts. Paunova-Krusteva, PhD

Applied Microbiology and Microbial Biotechnologies Chairman: Prof. M. Angelova, DSc Vice Chairman: Assoc. Prof. A. Gusterova, PhD Secretary: Assist. Prof. N. Kostadinova, PhD

Pathogenic Microorganisms and Infectious Immunology Chairman: Prof. T. Vassilev, DSc Vice Chairman: Corr. Member of BAS H. Najdenski, DSc Secretaries: Prof. N. Ivanovska, DSc; Assoc. Prof. V. Kussovski, PhD

General Virology Chairman: Acad. A. S. Galabov, DSc Vice Chairman: Assoc. Prof. L. Doumanova, PhD Secretary: Assist. Prof. V. Nikolova, PhD

*Carried out in the Stephan Angeloff Institute of Microbiology, BAS

ANEX 1

Press Conference of the SAIM-BAS (November 12, 2015) in Bulgarian News Agency (BTA): “Barriers to the best microbiological institute in South-East Europe”. The former Director Acad. Angel S. Galabov, current Director Corr. Member Hristo Najdenski and scientists from the Institute expressed their opinion about the problems concerning research funding

The consecutive 9th Balkan Congress of Microbiology, Microbiologia Balkanica’2015, was carried out in Thessaloniki (Grece) at October 22-24, 2015. More than 230 is the number of registered participants, 53 of which young scientists. SAIM - BAS was presented by 28 scientists, giving 4 plenary lectures, 12 oral presentations and 15 posters.

Celebration “Day of the Stephan Angeloff Institute of Microbiology”, March 14, 2015. The Director - Prof. Hristo Najdenski, Corresponding Member of BAS congratulates Prof. Lydmila Boyanova, MD, DSc, from Medical University of Sofia presenting a very intriguing lecture entitled “Helicobacter pylori – the carcinogenic bacteria”.

The President of the Foundation “Acad. Prof. Dr. Stephan Angeloff”, Acad. Angel S. Galabov, deliveres the traditional annual prize for the best scientific publication by young microbiologist to Tsvetelina Paunova-Krusteva, PhD from The Stephan Angeloff Institute of Microbiology, Department of General Microbiology.

Scientific Symposium „Beneficial and pathogenic microbes for healthier life and safety foods”, April 8-9, 2015, Sofia. Chairpersons: Assoc. Prof. Svetla Danova, DSc and Corr. Member Hristo Najdenski, DSc

Scientific Conference with international participation “The Antibody repertoire as a biomarker”, September 23, 2015, Sofia (BG09 PROGRAMME EEA SCHOLARSHIPS FUND). The chairman of the Organizing Committee is Assoc. Prof. Anastas Pashov, PhD.

2nd International conference on “Natural products utilization: from plants to pharmacy shelf”, October 14-17, 2015, Plovdiv, Bulgaria. The chairman of the Organizing Committee is Assoc. Prof. Milen Georgiev, PhD.

EDITORS Professor H. Najdenski, DVM, DSc, Corr. Member of BAS Professor M. Angelova, DSc

TECHNICAL ASSISTANT Vessela Yankova