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Absztraktfüzet Magyar Mikrobiológiai Társaság és az MMT Alapítványa www.mmt.org.hu A Magyar Mikrobiológiai Társaság 2012. évi Nagygyűlése ABSZTRAKTFÜZET Helikon Szálló, Keszthely 2012. október 24-26. MONITORING THE BIOGAS PRODUCING ARCHAEA COMMUNITY VIA MOLECULAR BIOLOGICAL METHODS N. ÁCS, Z. BAGI, ETELKA KOVÁCS, R. WIRTH, K.L. KOVÁCS Department of Biotechnology, Faculty of Science and Informatics, University of Szeged, Szeged, Hungary Biogas is one of the most promising renewable energy carriers since it has got the potential to produce energy from numerous biomasses, combining the production of green energy with the elimination of hazardous waste streams. Biogas, the main product of the anaerobic digestion (AD) process can be utilized by burning in combined heat and power (CHP) units or after purification it can be treated equivalently to the natural gas. If we want to enhance the produced biogas yield, or digest unconventional substrates, a deeper understand of the microbial community involved in the fermentation is requisite. This can be achieved with the culture independent methods. These molecular biological tools (TGGE, DGGE, ARDRA, RISA, T- RFLP) are widely applied today. High protein content substrates are considered toxic for the biogas fermentation due to the high level of ammonia released upon protein degradation. In this study systematic experiments were conducted in 5 litre CSTR (continuous stirred) fermentors, fed wit protein-rich substrates (casein or pig blood). We selected T-RFLP technique to determine the abundant archaeal microbial groups from the assortment of genomic DNA in the fermentation sample. Fluorescent labeled primers targeting the large subunit of the methyl-coenzyme-M-reductase (mcrA) gene were used to generate the mixed PCR product from the purified gDNA. The restriction endonuclease, RsaI was applied in order to generate the RFLP patterns of the selected samples. We concluded that the unconventional substrate source affected not only the primary consumers (hydrolyzing bacteria) but the archaeal community as well. A significant shift could be observed, confirming the dominant role of the genus Methanoculleus in the conversion of protein-rich material into biogas. The publication/presentation is supported by the European Union and co-funded by the European Social Fund. Project title: “Broadening the knowledge base and supporting the long term professional sustainability of the Research University Centre of Excellence at the University of Szeged by ensuring the rising generation of excellent scientists.” Project number: TÁMOP- 4.2.2/B-10/1-2010-0012 SURVEILLANCE OF HUMAN ROTAVIRUSES IN 2007-2011, HUNGARY 1 1 2 3,4 5 6 7 BRIGITTA ANTALNÉ LÁSZLÓ , J. KÓNYA , JUDIT DEÁK , F. JAKAB , ÁGNES JUHÁSZ , ILDIKÓ SÁNTHA , K. BÁNYAI AND THE HUNGARIAN ROTAVIRUS SURVEILLANCE NETWORK 1Department of Medical Microbiology, Medical and Health Science Center, University of Debrecen, Debrecen; 2Department of Clinical Microbiology, University of Szeged, Szeged; 3Virological Research Group, János Szentágothai Research Center; 4Institute of Biology, Faculty of Sciences, University of Pécs, Pécs; 5Microbiology Diagnostic Laboratory, Laboratórium Ltd., Hajdú-Bihar County, Debrecen; 6North Hungarian Regional Institute, Hungarian National Public Health and Medical Officer Service, Miskolc; 7Institute for Veterinary Medical Science, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary Group A Rotaviruses are the major viral pathogens causing acute dehydrating gastroenteritis among children less than five years of age. The availability of rotavirus vaccines has resulted in an intensification of post vaccine strain surveillance efforts worldwide to gain information on the impact of vaccines on prevalence of circulating rotavirus strains. In this study, the distribution of human rotavirus G and P types in Hungary is reported. In addition, the VP4 and VP7 genes of G1P[8] strains were sequenced to monitor if vaccine-derived strains were introduced and/or some strains/lineages were selected against. The study was conducted in 8 geographic areas of Hungary between 2007-2011. Rotavirus positive stool samples were collected from diarrheic patients mostly <5 years of age. Viral RNA was amplified by multiplex genotyping RT-PCR assay, targeting the medically most important G and P types. When needed, sequencing of the VP7 and VP4 genes was performed. In the study period, a total of 2380 rotavirus strains were successfully genotyped. Our results indicated the dominating prevalence of genotype G1P[8] (44.87%), followed by G4P[8] (23.4%), G2P[4] (14.75%) and G9P[8] (6.81%) strains. Uncommon strains were identified in a low percentage of samples (4.12%). Phylogenetic analysis of 318 G1P[8] strains identified 55 strains similar to the Rotarix strain (nt sequence identities; VP7, up to 97.9%; VP4, up to 98.5%) although their vaccine origin was unlikely. Current vaccines would have protected against the majority of identified rotavirus genotypes. A better understanding of the potential long-term effect of vaccine use on epidemiology and evolutionary dynamics of co-circulating wild type strains requires continuous strain surveillance. MOLECULAR PHYLOGENETIC RECONSTRUCTIONS WITH A DISCRETE MATHEMATICAL METHOD, THE BOOLEAN ANALYSIS ESZTER ARI Institute of Population Genetics, University of Veterinary Medicine, Vienna, Austria; and Department of Genetics, Eötvös Loránd University, Budapest, Hungary Boolean analysis, method for sequence comparison uses the Iterative Canonical Form of Boolean functions [1, 2]. It considers sequence information in a way entirely different from standard phylogenetic methods (i.e. Maximum Parsimony, Maximum- Likelihood, Neighbor-Joining, and Bayesian analysis). The performance and reliability of Boolean analysis were tested and compared with the standard phylogenetic methods, using artificially evolved – simulated – nucleotide sequences 1 and the 22 mitochondrial tRNA genes of the great apes. At the outset, we assumed that the phylogeny of Hominidae is generally well established, and the guide tree of artificial sequence evolution can also be used as a benchmark. These offer a possibility to compare and test the performance of different phylogenetic methods. Trees were reconstructed by each method from 2500 simulated sequences and 22 mitochondrial tRNA sequences. Considering the reliability values (branch support values of consensus trees and Robinson–Foulds distances) used for trees produced by different phylogenetic methods, BOOL-AN appeared as one of the the most reliable method. It can be concluded that Boolean analysis is a promising alternative to existing methods of sequence comparison for phylogenetic reconstruction and congruence analysis. Therefore this method can also be applied for inferring phylogenies of taxa with ambiguous evolution, like non-marine planktonic picocyanobacterial isolates (Synechococcus and Cyanobium). I investigated their phylogeny (based on the 16S ribosomal RNA genes) both with BOOL-AN and standard phylogenetic methods. [1] Jakó É, Ari E, Ittzés P, Horváth A & Podani J (2009) Mol. Phylogenet. Evol. 52(3): 887-97. [2] Ari E, Ittzés P, Podani J, Thi QC & Jakó É (2012) Mol. Phylogenet. Evol. 63(1): 193-202. FUROCOUMARINS: A NEW, POTENTIAL GROUP OF HIV-1 INHIBITORS? 1 2 3 4 5 1 ÉVA ÁY , NÓRA TAKÁCS , ÁGNES GYURIS , KATALIN KELEMENICS , JUDIT HOHMANN , J. MINÁROVITS 1Microbiological Research Group, National Center for Epidemiology, Budapest; 2Department. of Applied Biotechnology and Food Science, Budapest University of Technology and Economics, Budapest; 3RT-Europe Research Center Non-profit Ltd., Mosonmagyaróvár; 4GalenBio Ltd., Mosonmagyaróvár; 5Institute of Pharmacognosy, University of Szeged, Szeged, Hungary Human immunodeficiency virus (HIV), the causative agent of acquired immunodeficiency syndrome (AIDS), is still one of the leading causes of death in low-income countries. In high-income countries, highly active antiretroviral therapy (HAART) improved the life expectancy of HIV infected individuals. Rapid replication, high mutation rate and frequent recombinations of HIV can contribute, however, to the development of drug resistant variants, which can replicate even in the presence of antiretroviral drugs, leading to the failure of therapy. Accordingly, development of new inhibitors is required. Usage of naturally occurring compounds probably could increase the applicability and availability of drugs also in low income countries, where the epidemic is the most considerable. The aim of this study was to examine the anti-HIV effects of three furocoumarin compounds from Ruta graveolens and Evodia hupenhensis. We studied the cytotoxicity and inhibitory effect of byakangelicin, 8-geranyloxy psoralen and izopimpinellin on MT4 cells by MTT cell proliferation assay in vitro. The antiviral activities were determined by p24 antigen assay. The results demonstrated that out of the 3 compounds tested only 8- geranyloxy psoralen had an inhibitory effect on HIV-1 replication. The mechanism of action of 8-geranyloxy psoralen remains to be elucidated. ANAEROBIC FERMENTATION OF DISTILLERY THIN STILLAGE Z. BAGI, N. ÁCS, ETELKA KOVÁCS, K.L. KOVÁCS Department of Biotechnology, Faculty of Science and Informatics, University of Szeged, Szeged, Hungary Industrial processes create large amount of waste which are difficult and costly to decontaminate and properly dispose off. The distillery waste of starch or sugar based ethanol production is a light brown liquid with high organic content.
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