microbimconf.univ.kiev.uamicrobimconf.univ.kiev.ua/abstractbook2014.pdf · . 1’2014 3 huwiage...
TRANSCRIPT
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CONTENTS/ SECTION «MICROBIOLOGY»
« »………………………………………………………...... PLENARY SESSION
………………………………………………………………...... Boyko N.V. MICROORGANISMS AND IMMUNONUTRITION: NOVEL PRACTICAL TRENDS OF SELECTION AND APPLICATION OF PROBIOTICS………………..................
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ORAL/POSTER PRESENTATION
................................................................... Adamchuk-Chala N.I. EFFECT OF SOIL HETEROGENEITY ON SOIL MICROCENOSES OF ALFA-ALFA ROOT ZONE INOCULATED BY MARKED POPULATION OF RHYZOBIUM MELILOTY.................................................................................................. Antonenko L.O., Klechak I.R. THE ACTIVITY OF OXIDATIVE AND CELLULOLYTIC ENZYMES TRAMETES FR... Baig M.H., Danishuddin M., Kaushal L., Khan A.U. BLAD: A COMPREHENSIVE DATABASE OF WIDELY CIRCULATED BETA-LACTAMASES............................................................................................ Emelyanova E.V. PRACTICAL ASPECTS OF USES OF THE PULSE ADDITIONS METHOD........... Faidiuk I.V. PHAGE P1 LYSOGENIC CONVERSION OF PHYTOPATHOGENIC BACTERIA..... Holubenko O., Akulenko I., Shemchuk T. DETERMINATION OF SCFAS IN FAECES SAMPLES USING GC/MS................
17 18 18 18 19 20 21 22 23 23 23 24 24 25 26
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Huwiage G.M. THE USE OF PROBIOTICS TO TREAT CHILDREN WHO SUFFER FROM ACUTE GASTRO-ENTERITIS DIARRHOEA IN ALGARABOULLI HOSPITAL-LIBYA......... Khrokalo L. METHANE PRODUCING BY ANAEROBIC BACTERIA SELECTED FROM ORGANIC WASTES..................................................................................... Moroz J., Skurnik M. MOLECULAR, GENETIC AND STRUCTURE STUDIES OF THE LIPOPOLYSACCHARIDE (LPS) AS VIRULENCE FACTOR OF YERSINIA BACTERIA................................................................................................. Nidialkova N. ., Matseliukh .V., Varbanets L.D. MEDIUM OPTIMIZATION OF AN ELASTOLYTIC PEPTIDASE FROM BACILLUS THURINGIENSIS V -7324 ..................................................................... Zaets I.E., Kukharenko O., Podolich O.V. , de Vera J.-P., Reva O.N., Kozyrovska N.O. DNA PROFILING OF THE POLYMICROBIAL KOMBUCHA COMMUNITY............ Zelena L.B. INFLUENCE OF APOPTOSIS-INDUCING TREATMENTS ON THE EXPRESSION OF GENES THAT ARE INVOLVED IN UBIQUITIN-PROTEASOME SYSTEM AND FATTY ACIDS SYNTHESIS IN YEAST.........................................
. , . BRADYRHYZOBIUM JAPONICUM 8
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PSEUDOMONAS BASIDIOMYCETES
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PS UDOMONAS SYRINGAE PV. TROFACIENS...........................................................................................
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STENOTROPHOMONAS MALTOPHILIA
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PSEUDOMONAS PANTOEA
VITEK 2 COMPACT.................................................................................................
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XANTHOMONAS VESICATORIA......................
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WI-FI ..... ., ., ., .,
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BACILLUS , .............................................................
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37 37 38 39 40 41 41 42 43 44 45 45 46 47
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BRADYRHIZOBIUM JAPONICUM ....................
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ALBIDUS EUPENICILLIUM ERUBESCENS................................................... . ., . .
STREPTOMYCES GLOBISPORUS 1912-HP7 – .................................
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.LACTOBACILLUS.................................... ., .
BRASSICACEAE AMARANTHACEAE ...........................................
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ESCHERICHIA COLI 25922........... ., ., .
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ALU - BACILLUS SUBTILIS LYS-42..........................
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48 49 50 50 51 52 53 54 54 55 56 57 58 58
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, , EXOPHIALA ALCALOPHILA GOTO ET
SUGLY....................................................................................................... ., ., ., .
BRADYRHIZOBIUM JAPONICUM
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BACILLUS
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ANDIDA ALBICANS STAPHYLOCOCCUS AUREUS......................................................................
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7404, ................ ., ., ., .
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59 60 61 62 62 63 64 65 66 66 67 68 69 69
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BACILLUS................................................................... ., .
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PSEUDOMONAS AERUGINOSA ATCC 9027.................................. . ., ., ., .,
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VITEK COMPACT-2
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70 71 72 73 73 74 75 76 76 77 78 79 80 81 81
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CHLOROBIUM LIMICOLA -8...............................................
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BACILLUS , ............................................................................
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PSEUDOMONAS .................................
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ERWINIA CAROTOVORA SUBSP. CAROTOVORA.......................................... ., .
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AZOTOBACTER CHROOCOCCUM AZOTOBACTER VINELANDII ........................................................................
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CEREVISIAE ..................................................... .
RHODOCOCCUS ERYTHROPOLIS
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SECTION «IMMUNOLOGY»
« »........................................................................... PLENARY SESSION
.............................................................................. Skivka L.M. IMMUNOGENIC CELL DEATH IN HEALTH AND DISEASES.............................
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ORAL/POSTER PRESENTATION
.................................................................... Degen A.S., Topol I.O., Kamyshny A.M. FEATURES OF AN EXPRESSION OF THE T-BET AND GATA3 TRANSCRIPTIONS FACTORS IN EXPERIMENTAL PATHOLOGY...................... Galkin O.Yu. OPTIMIZATION OF CONDITIONS FOR THE ISOLATION AND PURIFICATION OF HUMAN IGE.......................................................................................... Kiyamova R. G. Kostianets .I., Dyachenko L.V., Lytovchenko A.S., Filonenko V.V. TUMOR-ASSOCIATED ANTIGENS AS MOLECULAR MARKERS FOR BREAST CANCER DIAGNOSTICS............................................................................. Nikulina V., Garmanchuk L., Senchylo N., Nikolaienko T. ADHESION POTENTIAL, PROLIFERATION AND THE GLUCOSE ABSORPTION LEVEL OF THE HELA CELLS UNDER THE INFLUENCE OF TEICHOIC ACID...... Palyvoda K.O., Oliinyk O.S., Lugovskaya N.E., Kolibo D.V., Lugovskoy E.V., Komisarenko S.V. GENERATION AND CHARACTERISATION OF RECOMBINANT SINGLE CHAIN VARIABLE FRAGMENT ANTIBODIES AGAINST PRO186-LEU197 PROTEIN C REGION.................................................................................................... Siryk G., Fedorchuk O., Malanchuk O., Skivka L. PHYSICAL ACTIVITY AT DIFFERENT DAYPARTS CHANGES CIRCULATING PHAGOCYTES FUNCTIONS.........................................................................
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IN VITRO.....................................................................
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SMUDGE- ....................................................................................
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IN VITRO IN VIVO .................................................................
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PSEUDOMONAS AERUGINOSA ARABIDOPSIS THALIANA ..................................
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SECTION «METHODICAL FUNDAMENTALS OF TEACHING OF MICROBIOLOGY AND IMMUNOLOGY»
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ORAL/POSTER PRESENTATION
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PLENARY SESSION/
Boyko N.V.
MICROORGANISMS AND IMMUNONUTRITION: NOVEL
PRACTICAL TRENDS OF SELECTION AND APPLICATION OF
PROBIOTICS Uzhhorod National University, Uzhhorod,
Ukraine [email protected]
Healthy life-style requires the con-sumption of well-balanced foods preferably rich in biologically-active compounds and fibre. The range of claimed healthy foods and functional products is enormous; but their role in health and wellbeing is still largely unclear. The possible way to substan-tially improve the human health is using of active dietary ingredients to mod-ulate normal gut microbiota and homeostasis of mucosal sites. We developed new conceptual criteria for the construction of pre- and probio-tics. In this paper we propose our background for the interpreting of “synbiotic” as a “complete and ba-lanced dietary intake of macronutrients and micronutrients beneficially affect-ing on microbial community structure”. Indexation of plants-originated priori-tised traditional foods from countries of Black Sea region (BSAC) had been performed within BaSeFood project1. The nutritional content, macro- and microelements, vitamins and folate had been determined and presented in EuroFIR composition data base. All prioritised foods and their major plant components are the greatest source of original beneficial microbes and novel prebiotic compounds. The ability of plants ingredients of national foods to selectively stimulate the commensal and to inhibit the potentially pathogen-
ic microbes had been confirmed in our in vitro and in vivo models. Recognition of the molecular mechanisms of directed modulation of gut microbiome and correspondingly human/host mucosal cross-talk following to the traditional foods intake will lead to effective regulation of host metabolic balance. A complex analysis of the traditional foods of the BSAC would make possible to select the potential candidates with putative functional properties due to their interaction with specific micro-organisms and present as an examples of “healthy immune-nutrition diet”. Our results support the concept of potential implementation of personalised diets for the patients with diet associated chronic diseases. 1The research leading to these results has received funding from the Euro-pean Community’s Seventh Frame-work Programme (FP7/2007-2013) under grant agreement N 227118.
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ORAL/POSTER PRESENTATION
Adamchuk-Chala N.I.
EFFECT OF SOIL HETEROGENEITY ON SOIL MICROCENOSES OF ALFA-ALFA ROOT ZONE INOCULATED BY
MARKED POPULATION OF RHYZOBIUM MELILOTY
Zabolotni Institute of microbiology and virology, Kyiv, Ukraine
[email protected] Soil biochemical properties respond most rapidly to small changes in soil, and directly related to soil organisms involved biogeochemical cycles of C, N and P as early indications of soil quality. Soil heterogeneity under different practices strongly influences soil microbial process, and an effect of management practices: the combina-tion of all operations, cropping prac-tice, fertilizer and other treatments conducted on applied to the soil may vary among different soil microcenos-es. im of investigation - evaluation of the impact of soil management on soil microcenoses used complex indices calculated by the combinations of different biochemical physic-chemical soil properties during formation of rhyzobial-legume symbiosis alfa-alfa plants inoculated by marked popula-tion of Rhyzobium meliloty 53Q30. In the studied soils , it was found that the presence of the genus Bacillus corre-lated with acidity and buffer soil acidity and phosphorus, the presence of the genus Pseudomonas correlated with indicators of soil electrical conductivi-ty, pH and buffer pH soil content of K, Ca i Al. The presence of the genus Rhyzobium dependent on the mechan-ical properties clay loam texture. Higher impacts of the vegetative indices of rhyzobial-legume symbiosis were selected in inoculated alfa-alfa
plants on the sandy loam, loam and clay loam soil textures, and moisture adversely to 23-55%. Research will be required to validate such approaches and to underprint the promotion of plant microbial linkages for improved N retention, and other ecosystem services, in managed system.
Antonenko L.O., Klechak I.R. THE ACTIVITY OF OXIDATIVE AND
CELLULOLYTIC ENZYMES TRAMETES FR
National Technical University of Ukraine "Kyiv Polytechnic Institute", Kyiv, Ukraine
[email protected] There is a sufficient amount of works aimed at studying oxidative and cellulolytic enzymes of Trametes fungi. The aim of our experiments was to study the activity of these enzymes in the Trametes fungi strains from the Collection of mushroom species of M.G. Kholodny Institute of Botany NAS of Ukraine Mycology Department where such studies had never been held before. The experiment showed the activity of extracellular monophe-nol monooxygenase and endo-1,4- -glucanase in 4 strains of different species (T. versicolor, T. hirsuta, T. ochracea, T. villosa) at different carbon and nitrogen nutrition sources. The cultivation was held under steady state conditions during 7 days at the temperature of 30° . The study of monophenol monooxygenase activity during its cultivation in media with different carbon and nitrogen sources showed that strain peculiarities are more important than suggested nutrition sources. The highest values of monophenol monooxygenase were marked for strain 1009 T. villosa – when starch was used as carbon source (102,5 units/cm3) that is twice more when glucose was used and 5
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times more when no carbon source was used at all). The high amount, in comparison to the other carbon sources, of extracellular protein also points at enzyme formation (37,7 mg/dm3) in culture fluid, part of which is being enzymatic. As for endo-1,4- -glucanase activity, its highest values were marked in 353 T. versicolor when fructose and mannitol were used, in 5302 T. ochracea when starch and maltose were used, in 5137 T. hirsuta on lactose and mannitol, in 1009 T. villosa on mannitol, starch and mal-tose. Thus, by changing carbon and nitrogen nutrition sources the strain 1009 T. villosa with the highest mono-phenol monooxygenase activity was selected and the variants of media were selected: with starch and pep-tone, with glucose and peptone. On the assumption of the highest endo-1,4- -glucanase activity 353 T. versi-color and 1009 T. villosa strains were selected for the further studies.
Baig M.H., Danishuddin M.,
Kaushal L., Khan A.U. BLAD: A COMPREHENSIVE
DATABASE OF WIDELY CIRCULATED BETA-LACTAMASES
Aligarh Muslim University, India [email protected]
Beta-lactamases confer resistance to a broad range of antibiotics and inhibitors by accumulating mutations. The number of beta-lactamases and their variants is steadily increasing. The horizontal gene transfer likely plays a major role in dissemination of these markers to new environments and hosts. Moreover, information about the beta-lactamase classes and their variants was scattered. Catego-rizing all these classes and their associated variants along with their
epidemiology and resistance pattern information on one platform could be helpful to the researcher working on multidrug-resistant bacteria. Thus, the beta-lactamase database (BLAD) has been developed to provide compre-hensive information (epidemiology and resistance pattern) on beta-lactamases. Beta-lactamase gene sequences in BLAD are linked with structural data, phenotypic data (i.e. antibiotic resistance) and literature references to experimental studies. In summary, BLAD integrates information that may provide insight into the epidemiology of multidrug resistance and enable the designing of novel drug candidates.
Emelyanova E.V. PRACTICAL ASPECTS OF USES OF THE PULSE ADDITIONS METHOD G.K.Skryabin Institute of Biochemistry and
Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow
Region, Russia [email protected]
Microbe cultivation in aerated sub-merged conditions has been used extensively in fermentations to pro-duce a variety of substances, cell protein and for transformation sub-stances into other more valuable products. The quantitative content for each nutrient should be properly chosen. The pulse additions method has been proposed by Dudina. It provides rapid identification of growth limiting component in the fermenta-tion. The presence or absence of the deficiency of any nutrients can be verified by the pulse addition of microdoses of nutrient components into the fermentation medium. In our studies the method of pulse additions was employed for the different nutri-tion components: substrates, which
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were the carbon and energy sources; various sources of nitrogen and phosphorus; so called "conservative substrates" as mineral components of the medium (Mg, Ca, Zn, Fe, Cu, Mn). The criterion for assessing was the fast change of the low inertial parameters of fermentation (pH of medium and oxygen consumption) in response to the pulse addition of an insignificant quantity of the growth limiting nutrition component into the fermentation medium. The value of response depended on the value of growth-limitation. The described above technique may be convenient and applicable both in fermentation microbiology and in biosensor analy-sis. This method - can be used for identification of culture growth limita-tion by nonconservative and also conservative substrates; - makes it possible to define multi-substrate limitation and at a high value of growth-limitation by other medium component to detect the deficiency of a nutrient, the concentrations of which in the medium are sufficient for non-limited growth; - is useful to estimate concen-tration of growth-limiting substrate by the response (the change of medium pH and oxygen consumption).
Faidiuk I.V.
PHAGE P1 LYSOGENIC CONVERSION OF
PHYTOPATHOGENIC BACTERIA Zabolotny Institute of Microbiology and Virology, NAS of Ukraine, Kyiv, Ukraine
[email protected] A comparison of the phage-host systems is essential for understanding the mechanisms of virus adaptation to variations of environmental conditions. Lysogenic conversion with the partici-pation of coliphage P1 in case of
uncommon bacterial hosts can expand our understanding of heterologous phage-bacterial systems.The purpose of the work was to investigate the peculiarities of P1 mc1ts100 lysogeny in Erwinia amylovora and Erwinia "horticola" bacteria associated with trees. 80% of used bacterial strains appeared to be susceptible to P1 infection which resulted in the cells’ phenotype conversion from CmS to CmR. The latter was proved to be caused by introduction of prophage DNA that is maintained as a single-copy plasmid of 94.8 kb in the cells. In the absence of selective pressure E. “horticola”(Eho) strains spontaneously loose CmR marker with a high frequen-cy up to 5.5%. It correlates with cells’ curing from the prophage DNA. Only in Eho 60-3m it occurs due to deletion in the region of Tn9 while the rest of replicon is kept. After the plasmid curing the characteristic value of phage sensitivity of cells is changed. In lysogenic cells the prophage genes of type III restriction-modification com-plex EcoP1I are actively expressed. The system formed by Eho 45 and 60 as well as their lysogenic derivatives and specific bacteriophages provides an opportunity to divide the latter into three groups according to the level of restriction in the course of their interaction with the enzyme EcoP1I. The difference in phage responses to the endonuclease presence in lyso-gens presumably correlates with the number of enzyme recognition se-quences and the adsorption sites availability. The expression of both immune and structural module of P1 is inappropriate or absent in lysogenized phytopathogens. Among all functions encoded by prophage efficiently expressed are solely the genes of
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mobile genetic elements: Tn9 cat-gene and RM-system EcoP1I. The constructed lysogenic strains allow for the exploration of RM-system Eco 1I interaction with polyvalent phages able to grow not only on E. coli, but also on phytopathogenic bacteria.
Holubenko O., Akulenko I., Shemchuk T.
DETERMINATION OF SCFAS IN FAECES SAMPLES USING GC/MS
Taras Shevchenko National University of Kyiv, ESC "Institute of biology”, Kyiv,
Ukraine [email protected]
Recently, an attention to the physio-logical changes of main microflora representatives is being rapidly increased. Thus there is a necessity of attracting faster and more effective methods to the assessment of micro-biocenosis. One of those techniques is gas chromatography–mass spectro-metry analysis of the short chain fatty acids(SCFAs) content in stool sam-ples, which are the major metabolic products of gastrointestinal tract microflora representatives. An applica-tion of such method is successfully used in global research institutes, but is still untapped in Ukraine. The purpose of our work was to study the process of sample preparation and SCFAs content analysis, to determine their spectra in stool samples of healthy rats and to compare our data with other sources. The research object was samples of faeces, which were taken from sexually mature rats(males) line Wistar, without abnor-malities of the gastrointestinal tract. All animals were kept in the NSC "Institute of Biology" vivarium at standard conditions. It is known when the pH of the fecal homogenate decreases, the output of SCFAs significantly increas-
es, therefore as the solvent was used 0.02 M hydrochloric acid. Analysis of investigated acids was performed by gas chromatograph Agilent Technolo-gies 6890N and capillary column DB_FFAP. As the mobile phase was used gas He. Quantitative and qualita-tive analyzes were conducted using mass-detector Agilent Technologies 5973 inert. In order to check the reproducibility of the conditions in each assay and determinate the concentration of acids, to each sample there was added an internal standard (4-methyl-valeric acid) with known concentration. We idetificated and calculated the concentrations of 5 carboxylic acids, which were pre-sented in all stool samples of healthy rats. These acids are acetic, propionic, butyric, valeric and iso-valeric. Processed technique of determining SCFAs in biological substrates using gas chromatography method has high accuracy, sensitivity, speed of data receiving, so can be used as screening method.
Huwiage G.M. THE USE OF PROBIOTICS TO TREAT
CHILDREN WHO SUFFER FROM ACUTE GASTRO-ENTERITIS
DIARRHOEA IN ALGARABOULLI HOSPITAL-LIBYA
High Institute of Polytechnique, Algaraboulli, Libya
[email protected] The term “probiotic,” which is derived from the Greek word for life, “bios,” was invented in the middle of the last century as a result of observing the beneficial influence of certain microor-ganisms on the intestinal flora. Recent-ly, for the correction of violations of the normal microflora of the human body, treatment dysbacterioses, many infectious and systemic human dis-
: . 1’2014
27
eases associated with microorgan-isms, are the most effective probiotics. The aim of study: The aim of our study was to determine wich probiotics are most effective in treatment of acute gastro-enteritis diarrhea in children. Materials and methods study: In our study we take two different series of commercial probiotic,wich is contain of different bacterial strains,these probiotc are Bifidombactrin (one dose contain contain 1 107 Bifidombacte-rium bifidum) and Lactobactrin (one dose contain 2 109 L.plantarum and L.fermentum) ,we also take 22 children as case study stay in Algarabolli hospital/Libya,age from 5 to 11 years and differ them to two groups, 11 child in each group, each child take one dose by three times aday. Results: After amonth of treatment and through an analysis showing that , agroup of children who have been given Lacto-bactrin, a single dose for three times a day improved their health and stop diarrhea quickly . While chldren who have been giving Bifidombactrin did not improved their heath. Conclusion: Microflora of the large intestine complete digestion through fermenta-tion, protect against pathogenic bacteria and stimulate the immune system. Probiotics can modify the composition and some metabolic activities of the microflora. whereas probiotics appear effective in treat-ment of childhood diarrhea, post-antibiotic diarrhea, and pouchitis. They affect immune modulation. The experimental researches will allow to establish what of widely used probio-tics are most biologically active and to give the recommendation on their use in a medical practice.
Khrokalo L. METHANE PRODUCING BY
ANAEROBIC BACTERIA SELECTED FROM ORGANIC WASTES
Kyiv National technical university of Ukraine “Kyiv polytechnic institute”, Kyiv, Ukraine
[email protected] Methane digestion is commonly considered as a three-stage process and provides due to activity of compli-cated anaerobic bacteria communities with hydrolytic, acetate-forming and methane-forming ones. Research was focused on screening of high methane productive anaerobic bacterial com-munity obtained from different organic wastes, growing and identification of methane-forming bacteria. The samples of wastes involved in experi-ment were taken from anaerobic sludge of waste water treatment station, pig manure, poultry excre-ments and fermented residue from biogas reactor. Storage bacteria culture was cultivated in strict anae-robic conditions (ORP less – 300 mV) and temperature + 35 0C on mineral liquid media (modified Zhilina’s media). Emissions of gases were analyzed on gas chromatograph LHM-8MD. Selection of methane-forming bacteria provided by passage of storage culture patterns on liquid media with addition of carbon source such as methanol and sodium acetate. Antibiotic amoxicillin in concentration 0,12 g/l was used for keeping pure cultures of methane-forming bacteria. Finally, the reinoculation of methane-forming bacteria was made on solid medium according to Hungate method in own modification. The purity of selection was controlled by microsco-py and gas emissions. Microslides were colored by Gram and viewed under light microscopy. Methane-forming bacteria identification was
: . 1’2014
28
provided by keys (Bergey, 2001; Iastremska, 1993). Bacteria communi-ties selected from poultry excrements and fermented residue from biogas reactor had the highest amount of methane production. Experiment results also demonstrated that activity of methane emission is larger in bacterial community patterns than in patterns with selected methane-producing bacteria cultures. Among methane-forming bacteria the strains of genera Methanosarcina and Metha-nothrix were identified. Further bacte-ria selection could be used for creation of bacteria enzyme drugs for applica-tion in biogas technologies. Fermenta-tion of organic wastes and biomass an industrial scale gives the possibility to obtain energy carrier, high-quality organic fertilizer and reduces human impact on the environment.
Moroz J., Skurnik M. MOLECULAR, GENETIC AND
STRUCTURE STUDIES OF THE LIPOPOLYSACCHARIDE (LPS) AS
VIRULENCE FACTOR OF YERSINIA BACTERIA
Haartman Institute, University of Helsinki, Helsinki, Finland
[email protected] Yersinia enterocolitica (Ye) is a well known human and animal pathogen. Among humans, the pathway of Ye associates with intestinal disease, such as enterocolitis, with inflammatory diarrhea, ileitis, mesenteric appendici-tis and gastroenteritis. Ye is a Gram-negative bacterium that contains the leaflet of its outer membrane a large number of lipopolysaccharide (LPS) molecules. LPS is a glycolipid consist-ing of three domains: lipid A moiety, the core oligosaccharide (OS), and the distal O-antigen (O-Ag) capping part.
The WaaL protein catalyzes the ligation of O-Ag onto the lipid A core. Based on in silico investigations, Ye possess 2 oligo-/polysaccharide ligases (WaaLos and WaaLps). The current studies were aimed to estimate importance of the ligases to control the LPS substitutions through construction of both deletion and catalytic mutants for virulence and structural studies. The mutagenesis of the waaLos and waaLps genes was aimed to be carried out using the allelic exchange strategy. That includes the use of suicide vectors which by two homologous recombination events exchange the wild type gene with a mutated gene. Suicide vectors were constructed for mutating both waaL-genes and were used to generate individual waaLos and waaLps mutants of Ye serotype O:3 and O:8. The isolated mutant candidates were confirmed by PCR-analysis. The LPS phenotypes of the Ye O:3 and O:8 waaLos and waaLps knock-out mu-tants were analyzed by DOC-PAGE and silver staining of proteinase K treated whole cell lysates. Hereby, obtained data show correlation between ex-pression of outer core and presence waaLos gene in bacteria genome. However, absence of waaLps gene, in waaLps single mutant, was compen-sated by the remaining WaaLos adequately. Further investigations with double mutant and complementation experiments will help to understand these mechanisms.
: . 1’2014
29
Nidialkova N. ., Matseliukh .V., Varbanets L.D.
MEDIUM OPTIMIZATION OF AN ELASTOLYTIC PEPTIDASE FROM BACILLUS THURINGIENSIS V -7324
Danylo Zabolotny Institute of Microbiology and Virology of the NAS of Ukraine, Kyiv,
Ukraine [email protected]
The medium optimization for the strains-producers cultivation to accumulate of the microbial enzymes has a great practical importance for industrial applications today. Besides to optimize the medium composition it is used either the empirical methods or a mathematical planning of the expe-riment. Previously it has been shown that Bacillus thuringiensis V -7324 synthesized peptidase with elastolytic activity that is a perspective for the medical application. So the aim of this study was optimization of the nutritious medium of B. thuringiensis V -7324 for the maximal accumulation of elastolytic enzyme. By means of the screening experiment it was estab-lished that all components are signifi-cant except a gelatin. A substitute of nitrogen source in the nutritious medium to methionine, valine, threo-nine, alanine and arginine led to an increase of specific elastolytic activity at the supernatant of culture fluid by 10.0-16.5 times. While an addition of ammonium sulfate and sodium nitrate resulted to increase of activity by 4.4 and 2.4 times respectively. A substitute of carbon source to an arabinose and rhamnose led to increase of specific elastolytic activity on 65-90 U/mg of protein. Due to a high cost of the amino acids it was chosen an arabi-nose and ammonium sulfate for further research to accumulate of B. thurin-giensis -7324 elastolytic pepti-
dase. Selection of the inorganic nutrition was shown that an zinc sulfate need to be replaced by zinc acetate in the basic medium for the accumulation of the investigated enzyme. An optimal ratio of selected carbon and nitrogen sources was found by a bifactorial experiment on three levels. It was established that the optimal concen-trations of arabinose is 13 g/l and ammonium sulfate – 14 g/l. Thus, the maximal accumulation of B. thurin-giensis -7324 elastase (132,5 U/mg of protein) is achieved on the nutrient medium having this composi-tion (g/l): arabinose – 13,0, (NH4)2SO4 – 14,0, 2 4 – 1,6, (CH3COO)2Zn – 0,25, MgSO4.
Zaets I.E. 1, Kukharenko O. 1,
Podolich O.V. 1, de Vera J.-P. 3, Reva O.N. 2, Kozyrovska N.O. 1
DNA PROFILING OF THE POLYMICROBIAL KOMBUCHA
COMMUNITY 1Institute of Molecular Biology & Genetics of
NAS of Ukraine, yiv, Ukraine 2 Bioinformatics and Computational Biology
Unit, Department of Biochemistry, University of Pretoria, South Africa
[email protected] High-throughput sequencing allows obtaining DNA barcodes from the environmental or man-made samples, allowing therefore the taxonomical assignation of microbial community members and the uncover of uncultiv-able microbial species, which cannot be detected by microbiological me-thods. In this work, we applied pyrose-quencing of 16S rDNA and ITS ampli-cons for the bacterial and yeast species profiling of the Ukrainian ecotype of the kombucha culture (UEKC), which was selected besides others as a cellulose-forming bio-
: . 1’2014
30
component for the international space exposure project BIOMEX (ESA). 454 pyrosequencing of appropriate barcodes and bioinformatics data analysis has been performed for the microbial species profiling of the UEKC. DNA samples were isolated from a 14 day liquid kombucha culture and a biofilm fraction of the culture separately. The DNA reads obtained from the sequencer were aligned locally by BLASTN. The BLASTN results were merged and visualized by MEGAN 4.67.4. The BLASTN output files were searched by an in-house BioPython-based script to retrieve the statistics of the top scored hits for all reads. It was found that UEKC com-prised two bacterial (Proteobacteria, Firmicutes) and yeast (Ascomycota) phyla, and several unknown pro- and eukaryotic microbial organisms. The core community includes acetobacte-ria of two genera (Gluconacetobacter, Gluconobacter) and yeasts (mainly, Saccharomycetes of Brettanomyc-es/Dekkera, Pichia, and Candida genera). The presence of several species, which previously had not been associated with the kombucha com-munity, was discovered that included Herbaspirillum spp., Halomonas spp. and several other occasionally occur-ring microbial organisms. Based on these data, the precise species inventory of microbial association will get us insight on the functionality of this community as whole entity. This in turn makes it possible to design model consortia of defined microbial organ-isms for a basic research and biotech-nology industry.
Zelena L.B. INFLUENCE OF APOPTOSIS-
INDUCING TREATMENTS ON THE EXPRESSION OF GENES THAT ARE
INVOLVED IN UBIQUITIN-PROTEASOME SYSTEM AND FATTY
ACIDS SYNTHESIS IN YEAST Zabolotny Institute of Microbiology and Virology, NAS of Ukraine, Kyiv, Ukraine
[email protected] Yeast cells are affected by different environmental stresses that can lead to cell death. This complex process is associated with changes in cell mem-brane and mitochondria structure, protein degradation, producing of abnormal proteins, chromatin frag-mentation, DNA breakage and others. In our study the expression of genes involved in fatty acids synthesis (ole1 and fas1)and ubiquitin-proteasome system (ubc6) after treatments with H2O2 and acetic acid was evaluated in two wild-type Saccharomyces cerevi-siae strains and their polyphospha-tase-deficient mutants. The level of gene expression in yeast cells was analyzed by RT-PCR. Comparative analysis of results obtained by RT-PCR revealed no changes of fas1 expres-sion in most cases with the exception of one wild-type strain and its mutants after treatment with H2O2 when gene expression was not detected. The expression of ole1 and ubc6 genes depended on strain and type of treatments. The level of their expres-sion raised in wild-type strains after each of treatments used. The increas-ing of ole1 expression was observed after acetic acid treatment in poly-phosphatase-deficient mutants of one strain whereas the slight decreasing or no changes was detected in the mutants originated from another. The treatment with H2O2 caused the increasing of ubc6 expression in most
: . 1’2014
31
cases. Thus, results of our research showed strain-specific mode of induction or repression of gene expression encoding ubiquitin-conjugating enzyme and two enzymes of fatty acids synthesis under influence of stress conditions in S. cerevisiae strains.
. 1, . 2
BRADYRHYZOBIUM JAPONICUM 8 634
1 , , ;
2 . , ,
- –
.
, 38–42% , 18–23% , 25–30% -, , ,
.
. -
. - Bradyrhizobium
japonicum.
Bradyrhizobium japonicum M8 634 , 8+634 . -
,
2012-2013 .: ( 8, 634 , 8+634 ) - (74,72,75);
(94,84,83); (163,132,133); (19,7,14,7,16,6) ,
-(70,8, 79, 120, 15,3).
,
, -,
, -
. -
,
. , -
-. ,
-
.
: . 1’2014
32
., ., .
,
. . ,
, --
.
, -
-.
-
,
, , -
.
-. ,
,
, -, -
-.
, 35-45°
Desulfovibrio, 60°
Desulfotomaculum Desulfomicrobium.
16S , -
2 100%
GenBank - Desulfovibrio sp. DSM
12803 (AJ251630.1), 3
92% - GenBank
Desulfotomaculum sp. ECP-C-5 (AF529223.1),
4 99% -
Desulfomicrobium baculatum DSM 2555 (AY464939.1).
-. -
-
, --
.
. ., . .
"
", , [email protected]
-,
.
: . 1’2014
33
,
. -
, -.
. -
( , , , ) -
( , , , , , ,
) . -
, -
(P. mirabilis, S. aureus MRSA, E. cloaceae, K. pneumoniae, P. aeruginosa); (S. enterica, Shigella dysenteriae, L. monocytogenes, EPEC E. coli)
(E. coli 058, E. faecalis,);
(L. acidophilus, L. delbrueckii, L. casei, L. fermentum, B. dentium).
, :
, , .
-
. in vitro -
-.
– -, -
. -,
, -
-
.
, .
. 1, . 1,
. 1, .
2 . 2
1 . ;
2 , ,
--
.
Purpureocillium lilacinum. . lilacinum Chaetomium aureum
(3,7x106-
3,7x108 ) . , ,
-
-
: . 1’2014
34
. -, -
.lilacinum
» -
. Purpureocillium lilacinum
-:
- ( 5%);
(0,02-
0,2%);
(0,002, 0,2%). ,
-
- (0,2%). -
« » 20%
». - « -
» « » 2% . ,
2O2 -
. -,
--
2%. ---
.
« »
P.lilacinum », , -
, -
.
., ., .
. . .
, ,
,
- Enterobacteriaceae. -
,
, .),
-
. -
. , ,
-. –
-.
- 30
: . 1’2014
35
.
. . -
t- .
, -
28,6% -
Candida (8,7% ; p<0,05) 14,3% – Enterococcus sp. (0% ; p<0,05).
--
60,9% Streptococcus pyogenes
(42,9% ; p<0,05), 30,4% - Staphylococcus epidermidis
(14,3% ; p<0,05), 17,4% -
Enterobacteriaceae (0% ; p<0,05).
St. aureus, Str. pneumoniae, Neisseria sp.
(p>0,05).
. ,
--
, -
– .
., .
», , [email protected]
,
.
, ,
-. ,
,
, -
.
. ( ,
, , ) ,
-. -
, -
. ,
-
( 1 ): 1 – , 25 - . , 15,4 – .
. , 19 – . .
100 -: 62 - . , 1,6 – . , 2,6 – .
, 2,2 – . -. ,
: . 1’2014
36
, -
100 3 ( .
), 4,6 ( . ), 2,4 ( . . ), 2,8
- . .
-
2,1 ( . ), 2 ( . ), 1,3
– . . , 5,8 – . . -
3,8 ( . ), 1,2 ( .
), 6,3 ( . ), 3,1 ( . -
). , -
, -, -
0
100 .
., .
PSEUDOMONAS BASIDIOMYCETES
,
Pseudomonas, Basidiomycetes,
, , .
, -
, .
-
.
Pseudomonas,
. -
(Agaricus bisporus (J.Lge) Imbach) – Sylvan 130 Hauser A-15.
(Lycoperdon perlatum
Pers.) , (Pleurotus ostreatus
Kumm.) -.
: -,
,
) ),
. -
-
) ( ).
-. ,
-
.
Pseudomonas (P.fluorescens (biotype G-syn. tolaasii))
-
: . 1’2014
37
. ,
, ,
, ,
.
, -
.
., . ., -. .
. .
-,
- ( ),
,
-,
.
-
Roundup Ready (RR) . -
Bradyrhizobium japonicum -6035 RR 40-3-2.
. -,
RR -
B. japonicum -6035, --
( ).
-
1,5 , -
1,2 .
3,9 , -
1,5 . ,
- RR
, -
1,5 – 2,8 . -
.
, , -
.
., .,
., .
PS UDOMONAS SYRINGAE PV. TROFACIENS
. . , .
[email protected] Pseudomonas syringae pv. trofaciens –
, -
. ,
,
: . 1’2014
38
.
-
, , . -
, ,
,
. -
. -
, -
( , 250 .), ( , 167 +
, 43 + , 250 .), ( 500 ), ( 60
), ( 15 , 700 .).
11 P. syringae pv. atrofaciens 4, 20, 912, 8462, 9010, 9057, 9400, 9404, 9417, 9747, 9780),
P. syringae pv. atrofaciens PDDCC 4394. -
P. syringae NCPPB
281. ---
, ,
. 24 -
28°
. , -
P. syringae pv. atrofaciens P. syringae. ,
10 ,
,
. , ---
P. syringae pv. atrofaciens P. syringae.
., .,
. LACTOBACILLUS
LANTARUM
. . , ,
,
. - L. lantarum,
-,
-
. L.
plantarum -, -
. -
, , , , ,
, , ,
: . 1’2014
39
109 L. plantarum. ,
12-48 ,
600 120 0 . 20% -
. L. plantarum
100 - 420 NaCl 2% - 8%, -
, 100 8% NaCl -
, .
51,4% - --
, 24% - 8,3% -
. L. plantarum
. -
. L. plantarum
. 10
L. plantarum, -,
, 2 ,
100 - 420 , 8% NaCl, -
,
-
.
. 1, . 2, . 1
1 . . , , ;
1 « », [email protected]
-
, -,
, , -
-
. -
.
-
.
-
.
, -
. ,
-
. «
» -.
: . 1’2014
40
200 , . -
,
17,4-32,1% . -
: -
- 2,2-3,7 ,
– 1,9-4,1 , – 1,1-1,4
.
68,6-80% 31,2-50%
.
. ,
,
-
. , -
.
. 1, 1., . 1, . 2
STENOTROPHOMONAS MALTOPHILIA
1
, , ; 2 .
, ,
. -
. , -,
.
-
, .
-
Stenotrophomonas maltophilia
.
Stenotrophomonas maltophilia. .
, Fe(II) Fe(III) -
. -
. ,
Stenotrophomonas maltophilia
. , -
. , 7
-,
11 .
Stenotrophomonas
maltophilia
: . 1’2014
41
.
7 .
Fe(II), 7 .
, , --
Stenotrophomonas maltophilia
-,
, Fe(II) .
, ., ., .
PSEUDOMONAS PANTOEA
VITEK 2 COMPACT
. . , . , 154, , ,
« » . .
, -
-, -
. -
VITEK 2 compact.
Pseudomonas Pantoea
- (GN
Colorimetric Identification Card).
Pseudomonas savastonoi pv. glycinea - 9074, 9072, 8541; Pseudomonas syringae pv. syringae 8414, 8570, Pantoea agglomerans -5113 , 8435, 8490, 9185.
Pseudomonas
. Pantoea agglomerans - -
.
Pseudomonas savastonoi pv. glycinea, Pseudomonas syringae pv. syringae Pantoea agglomerans,
. , Pseudomonas
savastonoi pv. glycinea 8541
Pseudomonas syringae pv. syringae.
.
, ., ., .
PSEUDOMONAS
. . , . , 154, , ,
« » . .
Pseudomonas
.
-
: . 1’2014
42
-
.
. .
VITEK 2
compact. -,
-,
--
. -
Pseudomonas – --
, -
. 15
--
28 . -
- VITEK 2 Compact
48 3 :
12 1, 9 , 12. --
Pseudomonas. ,
- Pseudomonas savastanoi
Pseudomonas syringae.
.
. . , , .
Fe( ) .
Fe(III) -
. -
( ) -
-.
-, -
.
( ) Fe(III) 0,5
. -
10 - – ,
, , , ( ).
-
1 .
- (2,6·102
),
: . 1’2014
43
.
. Deception (2,6·106 ).
1,6·103-1,1·104
. ,
) -
3,4·104 ( – ) - 5,9·104
). , -
-
, ,
.
. -
--
.
., ., ., .,
., .
, ,
, -,
-
.
-
. ,
-
. -
(180-230 ). ( ) (50 , .)
14 .
0,1 6%- .
®" (0,16 , per.os.) 14
8 .
, 14- 56- 9-
. --
. 1- 14-
. 56
Clostridium spp. (lg 3,5+0,3 2), (-) E. coli (lg 4,4±0,2 2), 2-3
Staphylococcus. Bifidobacterium, Lactobacillus, Propionibacterium .
-
56 .
(185,2±215,1 86,1±83,77 2)
: . 1’2014
44
56 . ,
. -, -
.
., ., .
( APSICUM ANNUM)
. . , ,
,
,
.
Xanthomonas vesicatoria Pectobacterium carotovorum
subsp. carotovorum. -
, -, -
. ,
,
. .
, -
. -,
, -
. , 15
, .
15 25 -,
- 25 . ,
14 -
( ),
X. vesicatoria, . -
- 25-35 . -
,
, 4 5 33-1;
22, X. vesicatoria 8 -
P. carotovorum. 33-1,
20-27 , 8 – 32 .
. , -
,
-
4 5 -.
: . 1’2014
45
., ., ., .
XANTHOMONAS
VESICATORIA
,
-. ,
-
quorum sensing. -
- Xanthomonas vesicatoria,
-.
.
- ( ) -,
(25%, 12%, 6%, 3%) (30
., 60 .)
--
BioTech. ,
X. vesicatoria <0,195%, .
- 1% 15 .
1 lg, 0,5% 30 . – 1,3 lg. ,
-.
-.
3% -
30 . 76,0%, 60 . – 65,4%; 6% – 66,4% 53,8% ;
12% – 56,5% 80,4% ; 25%
– 67,7% 82,4% -. 3%
30 . 58,1%, 60 .
– 75,7%; 6% – 76,7% 78,4% ;
12% – 87,4% 77,7% ; 25%
– 71,4% 70,8% -. , -
--
Xanthomonas vesicatoria. ., .
PHOTOBACTERIUM
PHOSPHOREUM
WI-FI
. . . , ,
- ( )
. -
: . 1’2014
46
Wi-Fi, --
.
Wi-Fi ,
-,
.
Wi-Fi --
. -
Photobacterium phosphoreum - 7071,
. -
, 3 10 ,
Wi-Fi TP-LINK TL-WR841N
(<6 , 100
2,4 ) 5,10 15 . -
-115. , Wi-Fi TP-LINK TL-WR841N
Photobacterium phosphoreum - 7071. -
. - Wi-Fi (5
) (
2-3%), 10 . 16 %. -
15 . .
38%. -
.
, -
Photobacterium phosphoreum - 7071
.
., ., ., .
., ., ., .
BACILLUS ,
, . ,
---
. -
.
.
- Bacillus
,
« -» « -». 13
Bacillus
: . 1’2014
47
.
, . -
1:10, 1:100, 1:1000 . -
LB -
. , ,
3 . ,
,
. --
(94-98% ).
4 : Bacillus subtilis 74,75,77, B. subtilis var.niger 83. -
B. subtilis 74 B. subtilis 77. ,
-
28% - B. subtilis 74 40%
B. subtilis 77 1:1000).
(1:1000) -
23%. , ,
,
, -.
. , .,
.
-
. . , ,
, 105 - 107
. 2 - 3 - 105
, 107 ( < 0,05),
- 105 .
--
105, ,
, - -
. --
: ; , -
. " -
", --
105 .
( ) 2- (49
%). -.
(33 %) (29 %). (13 %) , (8 %) -
(7 %). -
: . 1’2014
48
, , 27.8 % -
, 63.2 % - , 9%
( ).
(31.8 %) -
. --
18% .
-
9% 13% . " -
": -,
.
.
,
.
. ., . .
TN5- BRADYRHIZOBIUM JAPONICUM
,
– .
,
.
Tn5- Bradyrhizobium japonicum
. - (Glycine max L.
(Merr.)) ,
B. japonicum: 646 ( , ), 604 ( ),
66 ( ), 21-2 ( -) Tn5-
646: 9-1 ( ) 113 ( )
--
.
, -
.
.
, -
Tn5- -,
20- -
646 21-2. 28-
-
Tn5- 113. , -
B. japonicum,
-
: . 1’2014
49
,
. -
-.
. 1, . 2,
. 2, . 2, . 1, . 2
NEUROSPORA CRASSA -
1 -.
. , , ; 1 . ,
–
– ---
, .
,
. ,
,
, -
.
-
– N. crassa.
--
– ( -), -
, - – -
(3- -(5Z,8Z,11Z,14Z)-
(3-HETE)) (18- -
(9Z,12Z)- (18-HODE)). Neurospora crassa 3-HETE, 18-HODE
. 5 50 -
. , 3-HETE
18-HODE, ,
– – .
5
.
– 3-HETE ,
18-HODE – . ,
N. crassa, -. -
, 3-HETE 18-HODE, -
.
: . 1’2014
50
., .
-L-
CRYPTOCOCCUS ALBIDUS EUPENICILLIUM
ERUBESCENS . .
-L-
L- , ,
, -, , .
-L-
-, , ,
,
.
,
.
-L-:
Eupenicillium erubescens Cryptococcus albidus.
- -L-
. E. erubescens C. albidus -
, - TS Toyopearl
HW-60 Fractogel DEAE-650-s Sepharose 6B
-L- , -,
-,
-,
. , -
C. albidus E. erubescens, -
-L- , -
. ,
-L- E. erubescens -
, C. albidus. -
E. erubescens C. albidus -
-1,2- , -
, , -.
., .
STREPTOMYCES GLOBISPORUS 1912-HP7 –
-
S. globisporus 4Lcp 7. 4Lcp 7
.
2,6-3,5 37-51 -
.
. -
: . 1’2014
51
: - 4Lcp 7
,
; -
; - 7-10 -
- 44-48 -
(260 ., 28° ); -
10% , -
: -, .
,
4Lcp 7 -
, - 14,5
14,8 . , --
- 13-14,3 .
-
- 45,76 51,8 . , -
- 37 41
. -
,
4Lcp 7
.
., .,
.
.LACTOBACILLUS
», ,
, -
.
, i i -
i . -
, ,
.
, . --
. – , -, ,
i , .
--
. --
.Lactobacillus
« ». t +37°
MRS (Himedia) - ,
, , ,
: . 1’2014
52
. - (0,2 ).
520 . -
Lactobacillus plantarum MTCC 2621(I ). 11
.Lactobacillus 3 .
(U/ml; ±0,001) 24 48
L. rhamnosus LB3 IMB B-7038 - (0,019 i 0,03); L. bulgaricus LB51 - (0,009 i 0,013l); L. delbrueckii subsp. delbrueckii DSM20074 - (0,017 i 0,03 l). L. plantarum MTCC 2621 0,024 0,051 U/ml,
. , -
-. -
--
.
., .
BRASSICACEAE AMARANTHACEAE
.
. , , [email protected]
- ,
, .
-, -
, ,
.
.
-
Brassicacea Amaranthaceae
( 3 4). -.
. .
-: Brassicacea ( -, )
Amaranthaceae ( ).
– ,
-. -
. , , -,
. - ( )
–
– . Brassicacea, . Amaranthaceae, ,
. -
. -
,
: . 1’2014
53
.
. 1,5-2 .
–
.
. -
. Amaranthus -.
. ., . ., . .
. . , ,
-
.
, -. -
. , ,
-
, -
. :
-. -
- 2
60 . -
-
.
- ( ). -
,
, - (7,2),
(8,0) 1,25 40 .
6,0 - -
40 80 . -
10% -
, 106±9,3 .
, -
10% ,
-
, - -
-
.
., ., ., ., .,
., .,
: . 1’2014
54
., ., .
. . , , [email protected]
-
,
,
.
Pseudomonas - P. cepacia NU-327, P. fluorescens ONU-328, P. maltophilia ONU-329, -
,
. . P. fluorescens ONU-328 P. maltophilia NU-329
-, P. cepacia NU-327 – -
.
Pseudomonas -
. -
30 60 -
“ -29”. -
--
, . -
, -
.
30 45-52 %. 60
75-90 %.
, ,
. ---
- --
, ,
--
.
., ., ., .
,
ESCHERICHIA
COLI 25922
-.
, . ,
-
2-10 -, . , ,
, .
: . 1’2014
55
,
- Escherichia coli
25922. - ( ) -
. 3-12%
60-250 15-120 .
-
. , E. coli
< 0,2%,
- 1,95 . - Ct 2,1×105 × , 6×106
× 1,57×103 × ; -.
E. coli 3% 15 .
4,5 lg; 3% 15 . - 3,39lg, 12%
– 3,8 lg; 125 15 .
1,3 lg, 250 15 . – 4lg.
4-7 E. coli
10 . 1,36 lg, - 2,15 lg, -
250 1 . 0,47 lg.
, , - E. oli -
,
.
., ., .
, ,
. -
. :
-,
-. -
. - 14
50 . - 72
. 6%
1% . - (0,16 )
per.os 14 2-
. 72 14-
III .
. -
( 1,7+0,4 5,8+ 0,3
0+0 4,4 2 -). -
3 , -
: . 1’2014
56
. -
. -, - -
. -
Shigella Salmonella, .
14 -
.
. : 1)
--
; 2) -
.
., ., .
ALU- BACILLUS SUBTILIS LYS-42
-, ,
, . -
, .
,
. ,
-,
. , -
,
. , -
,
. -
,
.
,
, Alu-,
.
- 10
Tris-HCl (pH 7.5) .
Alu- Bacillus subtilis Lys-42
. -
60 .
, Alu-
-
. -,
Tris-HCl . -
-.
40 B 0.8%- -.
, -.
: . 1’2014
57
Alu- Bacillus subtilis Lys-42.
.
,
70
« » ( .
/ . .
; .: ., .,
. .; . .: ., .
– .: , 2013. – 808 .). -
, ,
. -,
, -
. 2- ,
,
. . (1873), ,
,
--
.
, 1-2 , -
: ),
), ( ), ( ),
), ),
), - ( ).
, ,
, , ,
1876 . -
, , 1877 – -
-,
1883 . -,
.
. , -
, , ,
.
2-3 .
. .
-, ,
, ,
: . 1’2014
58
-
.
,
,
. -
): ( – Agrobacterium radiobacter-204,
), ( – Enterobacter nimipressuralis-
32-3, ) - –
, , Bacillus
polymyxa ).
, -
(Lolium perenne L.) (Medicago sativa L.).
.
0-30 30-60 .
-, , -
,
. -,
, -
1,8-4,1 1,2-
2,2 . ,
,
.
2-4 1,5-2 . ,
, ,
. , -
.
-
.
., ., ., .
. . , ,
, -,
,
.
-,
. -
-,
. ,
: . 1’2014
59
, -
. --
P. eruginosa, ,
-
.
. -
, -
,
-.
--
. -
. , -
- 24 48
.
. -
--
.
., ., .
. . , ,
--
,
.
(II) (
--
). (
) -.
-
. ,
(II),
0.25 .
, , ).
13 ,
( 1 ). 1-4
(HiMedia Nutrient Agar, 800 ),
5-13 - ( 10% HiMedia
Nutrient Broth, 60-80 ). -
: . 1’2014
60
) ( )
1 4 – 100 , 3 – 200 ).
), (20-30
). ( ) 3-4
-
. , 15% - ) -
-,
. ,
,
(II),
.
., .
, ,
EXOPHIALA ALCALOPHILA GOTO ET SUGLY
, ,
Exophiala -, ,
, -
, , -.
Exophiala -
, .
. ,
-
, -
. ,
Exophiala.
, ,
Exophiala .
, ,
, , , , , , , ,
, ( , ,
) E. alcalophila FCKU 304
« ». -
, E. alcalophila, -
MEA ( alt-extract agar), --
, .
30 28±1° . -
, , -,
E. alcalophila ). -
, , -
, , , -, , .
E. alcalophila, -
.
: > > >
> > > > .
: . 1’2014
61
., ., ., .
BRADYRHIZOBIUM JAPONICUM
, ,
. --
, -, -
-
. -
,
.
.
(Glycine max (L.) Merr.) -,
(646) (604 ) B.japonicum
. . -
Agilent 2100 Bioanalyzer System (Agilent Technologies, Waldbronn, Germany).
,
-
, ,
, -
.
.
, -
B. japonicum, -
.
-, ,
, -
– , -
.
., ., ., .
BACILLUS
. . , ,
-
, -
. -
-
: . 1’2014
62
-
. ,
Xantomonas campestris pv. campestris 8003 -
B.subtilis 5/6, Bacillus sp. 41 Bacillus sp. A1, Pectobacterim carotovorum sabs. carotovorum
-1095 – Bacillus sp. 41.
: Agrobacterium tumefa-ciens 8628 B.subtilis 5/6 Bacillus sp. 41; P.carotovorum sabs. carotovorum -1095 – -
B.subtilis 5/6, Bacillus sp. 23/2; Clavibacter michiganensis 102
B.subtilis 5/6; X.campestris pv. campestris 8003 – Bacillus sp. 23/2. -
Pseudomonas syringae -1027
P.fluorescens 8573 - Bacillus
-.
, 2 (B.subtilis 5/6 Bacillus sp. 41)
-
, – Fusarium
graminearum 9G Cochliobolus sativus 10Z. , -
-
Bacillus. ) -
(15-51 × -1) Bacillus,
. , -
, -
(B.subtilis 5/6 Bacillus
sp. 41), -
-
.
. ., . .,
. ., . .
ANDIDA ALBICANS STAPHYLOCOCCUS AUREUS
, ,
[email protected] andida albicans
Staphylococcus aureus 20 % -
, , ,
27 % -.
, -
-. -
–
.albicans S.aureus.
. - 15 -
.albicans S.aureus,
. .albicans 885-
653 S.aureus 25923. ( ) -
-.
-
: . 1’2014
63
. . -
-
: – 32 , - – 5 ,
– 12,5 . -
.albicans S.aureus.
(0,0451±0,007) . , – 2,6
. . , -
.albicans S.aureus, – 2,6 . -
.
.
-
. ,
-,
, . ,
, , , -
, , , -
,
-. , -
-
,
. ---
5 . , ,
. -
----
. , -
.
.
, . , -
, .
, -
, -.
.
5,5-11,4.
, -
. -,
0,9 % -
: . 1’2014
64
, ,
. --
.
., ., ., .,
, ., .
® »
( ) – , -
. -
« ® » ( )
. : -
(m=180-230 ). 5-14
(50 , ). (0,16 , ) 4
)
14 . 1, 14 56
. (MDA) -
-
; (S D) -
; Egr-1 Sp-1 .
. : 1- ,
- (4,5 ) SOD (1,1
), MDA (2,4 , <0,05), Egr-1 (1,5
, =0,02) Sp-1 (1,2 , =0,04)
. - 2,6 MDA,
4,6 ( <0,01)
SOD. -
2 -
( ) (Clostridium spp., Staphylococcus aureus),
Bifidobacterium, Lactobacillus, Propionibacterium .
14 56
.
-, -
(3,6 ; =0,02)
. --
56 . :
.
: . 1’2014
65
., .
, ,
, 2,4- N-
. . ,
, .
, -
--
. -
. -
, : ( ,
) -. -
, -, ,
2,4- n-. -
.
, -
( --
),
. -
dI/dt ( ),
-. -
. Methylopila musalis
B-2646 . Alcaligenes xylosoxydans
subsp. denitrificans TD 2 -
. Chelativorans oligotrophicus LPM-4
-
. Rhodococcus erythropolis HL PM-1
2,4-. mamonas
testosterone BS 1310 (pBS 1010) n-
.
., ., .
. , ,
-.
-,
.
,
: . 1’2014
66
.
, ,
.
, ,
, , .
, --
. , .
. , -
-.
-
. ,
. ,
.
-. ,
, -
. ,
-
, .
. ., . .
. . , ,
. -
, -
. ,
-
: , -, , , -
.
. 7 -
Bacillus Bacillus subtilis
-7023, -
- Pseudomonas syringae pv. syringae 8511, Pseudomonas fluorescens 8573, Erwinia carotovora
: . 1’2014
67
8982, Xanthomonas campestri 80038, Clavibacter michiganensis subsp. michiganensis 102, Agrobacterium tumefaciens 8628.
--
. , Bacillus -
, -
. B. subtilis -
- Bacillus subtilis -
7023 Agrobacter
tumefaciens 8628, -
. -
Bacillus megaterium 2 B. pumilus 3.
, -
Bacillus subtilis -7023, -
.
., .
-
-
. , --
. ,
, .
Bradyrhizobium japonicum . -
(Glycine max), -
B. japonicum, 646 604 .
-,
. -
, -.
«Agilent
GC system 7890 » ( ) - 5975 . -
, , ,
, , 20-40 %
-,
-.
, -,
-
: . 1’2014
68
. -
, --
.
,
--
.
., ., ., ., .
EFSA
« », ,
[email protected] European Food Safety Authority (EFSA)
, . EFSA 2003
. EFSA , . -
. « -
», -
, --.
, -,
.
, -
-. , --
). ,
« » . -
, ,
. EFSA -
“ ” “ ”,
: (
). -
,
, -, , -
,
, --
.
. .
BACILLUS AMILOLIQUEFACIENS -7404,
. . . , ,
– -
: . 1’2014
69
.
- Bacillus,
.
- Bacillus
amiloliquefaciens –7404 Fusarium
graminearum Bipolaris sorokiniana .
B. amiloliquefaciens –7404 -
.
. ,
.
-
. --:
0,25% – ;
15% –
. ,
, , -
,
-. , , -
5 .
1 2.
. -
B. amiloliquefaciens –7404 ,
1 2. -
,
9,5 10,5 F. graminearum B. sorokiniana
.
., ., ., .
. . , ,
-, -
. --
- Meloidogine incognita -
in vitro -
0,5; 1; 2; 3; 4 24 . , 79
- 30
, 24 .
. ,
:
: . 1’2014
70
Cochliobolus, Alternaria, Fusarium
ladosporium – , , (
11 14 )
Clavibacter michiganensis, Pseudomonas syringae pv.
oronafaciens, P. syringae pv. trofaciens, P. savastanoi pv. glycinea,
P. corrugata, Xanthomonas campestris pv. vesicatoria, Pantoea agglomerans, -
- (
20 28 ).
-,
. ,
, -.
-
16S rRNA 1461 . ,
99,0 % - Amycolatopsis orientalis
AJ400711).
-. , -
, 4 « -,
». -
, Actinomycetales, Pseudonocardiaceae, Amycolatopsis, Amycolatopsis orientalis.
. .
, ,
, -
, .
–
-,
--.
, -
-
. ,
-,
,
. , -
. 0–30
25-28 %.
: . 1’2014
71
10–20 , 23–26 . ./ . -
, , 0–30
10-12 %. -
10-20
12-19 %.
-
. , 0-10 -
20-28 %, 10-20 - 25-32 %, 20-30 - 17-24 %. -
- 25-28 %,
12-19 %, - - 17-28 %.
, .,
., .
PSEUDOMONAS AERUGINOSA
, ,
[email protected] Pseudomonas aeruginosa –
-,
--
, . ,
, P. aeruginosa -
.
Escherichia
coli -906, Staphylococcus aureus -918, Micrococcus luteus Ac-469 -
Pseudomonas aeruginosa B-900. -
E.coli, S.aureus, M.luteus
P.aeruginosa 1:1. 1 .
-. -
(OriginLab 8,0). , P.aeruginosa -
-
(R=0,970). E.coli
14,71%,
(R=0,976). S.aureus
P.aeruginosa 33,8%; 50%
25%. -
- (R=-0.5),
, , ,
. M.luteus -
P.aeruginosa 11,5%.
(R=0,88), .
:
(R=0,976),
: . 1’2014
72
- 139%.
-
Micrococcus -.
., .
«
», , [email protected]
-
, -, -
.
,
-.
–
-,
(Glycine max Moench).
,
30 , 60 , 90 100 2 15 ,
30 , 45 100 2, (Glycine max
Moench.), .
-
: – ( ), -
– , – -
( ), - – , -
– . -
,
-.
.
,
30 2
70% 78,3% -
. .
,
, --
.
. ., . ., . ., . ., . ., . .
. . . , ,
(NNIS)
: . 1’2014
73
, - 16%
. -,
, , .
,
. :
,
. -
, , -,
S. aureus (n130) (n120). -
. ,
( 1,10±0,86 ) S. aureus; 4,53±2,03 – E. coli. -
, S. aureus ( 1,45±0,81 );
E. oli 5,88±2,62 ).
- S.
aureus ( 31,5±4,3 ); E. coli – 20,20±10,41 .
- S. aureus
8,04±4,24 ; E. coli – 14,93±14,47 , -
-.
, -,
-.
., ., ., .
.
, ,
-
, -
. --
« » --
– -
). ,
(n=210 ) 2+ Fe2+ -
) - 16 –26 %. -
Rhodococcus erythropolis -7277 Dietzia maris -7278
17,6 %, Gordonia rubr pertinct
-5005 Rhodococcus erythropolis -7012 – 13,6 %
. R. erythropolis
-7277 D. maris -7278 «Biot c»
) 30 1,0 %
4
: . 1’2014
74
2+ Fe2+ 1 -, 0,7
., – 500 ., – 1,5
2
, . ,
1000 5,4 109,
2,0 , – 87,8 %. -
48
0,5 % 5 %
. pH
6,8-7,2. -
« -»,
,
.
., .,
., ., .
;
, ,
-
,
.
-. :
, -
,
.
-.
50 .
-
, --
. 3-
4 5 3 .
100% :
80% ,
107 /1 .
-: Candida albicans,
Staphylococcus saprophyticus, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Enterobacter aerogenes, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Citrobacter diversus. 80%
: . 1’2014
75
-,
(
106-107 ). -
, -
, , 102-
103/ . ,
.
., ., .
. . , ,
-.
( -, ), ,
, --
.
.
-.
--
-. 6
, -
( ) ( ). -
.
- –
70-120 . , 10 -
-.
-.
, -,
,
. ,
.
., ., ., .
BACILLUS . .
, , -
. Bacillus
, -
. -
. , , 4-
,
: . 1’2014
76
B. subtilis -7023,
. ,
,
- B. subtilis -
7023 . 12 ,
: B. megaterium (3), B. subtilis
(3), B. cereus var. mycoides (3), B. pumilus (3). -
280 (240 ) 48
-,
. ,
-
B. megaterium 2,1 (67,7 – 67,3 ).
B. megaterium 9 -
. - B. subtilis 11
43,7 , B. subtilis -7023 – 31,7 , B. subtilis 13 –
16,3 . B. cereus v. mycoides . 10 16
20 , B. cereus v. mycoides .14 – 13,6
. C B. pumilus
B. pumilus 7, 55,2 -
. -
B. pumilus 3 4 24,3 15,3 .
, - Bacillus
-,
-
, .
., .
. .
--
, -
. --
-,
. ,
, -
(P. syringae pv. atrofaciens) (Xanthomonas translucens) -
. P. syringae pv. atrofaciens, ,
.
Pseudomonas -,
-. -
P. savastanoi pv. glycinea (
) X. axonopodis pv. glycines ).
P. syringae pv. tabaci Curtobacterium flaccumfaciens pv.
flaccumfaciens.
: . 1’2014
77
, P. syringae, Pectobacterium carotovorum subsp. arotovorum, Pantoea gglomerans, C. flaccumfaciens. ,
Clavibacter
michiganensis subsp. michiganensis,
. C.
michiganensis subsp. michiganensis
14- 18.
– P. xanthochlora - P. marginalis
16S .
,
Pectobacterium, Xanthomonas Pseudomonas.
P. syringae
, . -
, --
.
. 1, . 2
: ,
1 ;
2 . . , ,
,
--
, --
, -. -
--
, -, -
,
. ---
, -
. -
, ), --
, -. 95%
, -
,
. --
- -,
,
. - –
, ---
, -
: . 1’2014
78
(
, , ).
--
( , ), -
, -
. -
, , --
-.
, .,
.
PSEUDOMONAS AERUGINOSA ATCC 9027
, ,
,
. ,
, -
. -
, -,
.
, -
, ,
.
Pseudomonas aeruginosa ATCC 9027.
. -
( ) -
0,2% - 25%. 3% 6,25% 15-120 .
10 . 6%
. ,
Pseudomonas aeruginosa 9027 12,5%. Ct
4,6×105 ./ . -
3% 15 .
2,5 lg, 6,25 % 15 . – 3,5 lg.
. - 5×108
3 ,
5,56 lg , 7 - 7,1 lg .
10 6% .
.
- .
: . 1’2014
79
, Pseudomonas
aeruginosa ATCC 9027 ,
.
. ., ., ., .,
,
. - ( )
) ---
, Clostridium difficili..
. , ,
. :
.
, 180-230 . 5 .
– (H2O per.os), -
(50 ) 14 , 14 -
, 14 (0,16 ).
( ) 72
. , 20-
.
Bifidobacterium, Lactobacillus,
Propionibacterium ,
.
( 1,7±0,4 5,8±0,3 lg 0 4,4±0,2 ).
Shigella Salmonella.
Clostridium,
. - 14
. : 1) 14
-
Clostridium ; 2)14- ( )
-.
. ., ., ., .
. . . ,
, -
,
: . 1’2014
80
, .
, --
9.048-89 .
-
» ( 1) -. -
. ,
-,
100%,
25% . -
) -
Chaetomium globosum Stachybotrys chartarum. ,
C. globosum
90%), S. chartarum -
-.
55 18
Zygomycota Ascomycot -
. 2005 .
10 , S. chartarum.
2010 . 29 14
, Alternaria Chaetomium.
-
2010 . - 31
15 , -. Alternaria
.Penicillium. .Ascotricha
, . -
, (43,75%)
-
2010 .
., ., .
« . . -
», , [email protected]
, - ( )
,
. -
, -
. 36 17-55 ( ,
, -
) . -
.
( -). -
(72,22 % ) (77,8 %).
55,6 % , -
: . 1’2014
81
– 50,0 %, - – 27,8 %. ,
E. coli (50,0 %) Klebsiella ssp. (11,9 %). 66,67 %
Candida. -
7,37±0,15 lg . ,
, (6,78 0,12 lg ),
(6,58 0,16), (3,87 0,16 lg ).
S. aureus (4,50 0,12 lg ) KON (4,48 0,18 lg ).
E. coli 2,19 0,13 lg , Klebsiella ssp. – 3,64 0,04 lg , Candida – 3,24 0,11 lg
. ,
, -
.
. 1, . 2
VITEK COMPACT-2
1 « . .
»; 2
« -», ,
. -
,
.
, -,
Vitek-2 Compact. - 40 38 .
. :
– 10, – 9, – 9,
– 6, – 3, – 1. : P. aeruginosa (19 ), . baumanii (7), K. pneumoniae (3), S. marcesc ns (2), E. aerogenenes (3), E. coli (2), E. faecalis (3), S. aureus (1).
P. aeruginosa -:
47,4 % , – 80,0 %, – 100%. -
, , , – 15,8%
; – 10,5%. P. aeruginosa -
– -, , -, , -
I – IV ( , ,
, , -, , ,
), ( - 5,3 % ),
(15,8 % ). 7 A.
baumanii 1
2 – .
: . 1’2014
82
(K. pneumoniae, S. marcesc ns, E. aerogenenes, E. coli)
- ( , , -, ) – 80% 100%,
-, 1
K. neumoniae,
.
, .
.
.
(CRO42-,
CO2+, NI2+, CU2+, HG2+)
-,
.
,
.
(CrO42–),
(Ni2+, Co2+) -
, . , ,
(Cu2+ Hg2+). --
- ( – 4030 ), -
( ).
- ( ).
(100 ) , Nutrient
Broth ( HiMedia Laboratories Pvt. Ltd.) -
. K2CrO4, Ni(NO3)2×6H2O,
CoCl2×6H2O, Hg(NO3)2 ). -
, -, -
. -
». ,
-. ,
2 - 3 -
-
(0,1 – 10 ). :
50 Hg2+, 500 Co2+ Ni2+, 1000 Cr(VI), 20 000 Cu2+
; 100 Hg2+, 500 Co2+ Ni2+, 1500 Cr(VI) 5000 Cu2+
-. -
-, « -
» .
, ---,
: . 1’2014
83
-
« » .
., .,
., ., ., ., .,
., ., .
”
,
-.
,
.
:1. (Cf) - 50 / , , 2.Cf " " (Sym) - 0,16 /
, 4 Cf ).
14 . --. -
.
: -
, NO
. 14- Cf
:
Clostridium (lg 0,5±0,1 2) -
E.coli . -
, ,
, -
( ). Cf
64%,
NO .
-
, . -
Cf Sym --
Clostridium. -
48%, .
, -
, -. -
.
: . 1’2014
84
.1, .2
®
1 . , ,
, ; 2
, ,
-,
.- . ,
( , ),
-,
.
- E ®
- Lactobacillus:
L.plantarum 20 L.casei 6, - Lactococcus lactis 4/6,
, Fusarium, -
Aspergillus, -.
, -
(E.coli, S. aureus, P.aerogenosa, S.typhimurium)
3 (
28 30 ), ,
®, 35 . E ®
, . E ®
A.candidus, A.fischeri 15 - 22 ,
L.casei 6, - A.flavus - 14 , L.plantarum
20, , .
A.pulvinus 27 ,
L.plantarum 20 L. casei 6 .
- A.avenaceus
F.moniliforme. (E ®)
: A.avenaceus - 12,5 , F.moniliforme, F.sulphureum
28 . , ---
, -, ,
, -.
E ® -,
,
. ®,
. ., .
, ,
--
, -, .
, , -
: . 1’2014
85
, , . -
: --
- ( ).
(« -
»), ( ») (
). - 40
, -, , -
. 10 ,
. -
-, ; . -
.
S.aureus (50%), E.coli (30%), P.auruginosa (40%), .faecals, K.pneumoniae ( 20%), P.vulgaris, C.albicans A. calcoaceticus ( 10%). 7 -
1,5 .
7 . -
,
, , . -
. , -
.
. . , -
-
,
.
., ., .
. , ,
, -. -
,
. -, , -
, , , , -
, , .
-, , -
, -.
– -,
, , ( « »,
, . ). -,
-.
: . 1’2014
86
Neisseria sp., Staphylococcus aureus, Candida albicans, Enterobacter sp., Escherichia coli. -
.
-.
108 . ./ .
18-24 37±10° . 3- .
,
Staphylococcus aureus Escherichia coli.
25 . -
Enterobacter Neisseria.
. ,
,
,
.
., ., .
" "
-
, ,
-,
. : -
« -»
. , ,
. -
, -, , ,
. ,
» - 8 -
.
-.
--
. ,
.
, -.
2-3
,
Clostridium. -
.
1-2 . -,
-. -
,
: . 1’2014
87
, , , -
, E.coli -
-,
3 , Staphylococcus aureus
-.
-
. , -
--
.
., ., .
–
. . .
-
-.
. (Sorhus arvensis L.)
(Taraxacum officinale Wigg.) -
22 , -
.
, .
,
-. , -
: (Convolvulus arvensis L.),
(Elytrigia repens (L.), (Equisetum arvense L.),
(Setaria pumila (Poir.) Schult.) -
– (Triticum aestivum, 93), (Avena sativa, ) (Glycine max ) ).
, -
,
. -
, , -
, , ,
Rhodosporidium diobovatum Newell & I.L. Hunter. 342 ,
, -
, Rhodotorula sp. 345
342 ,
Rhodotorula, .
,
, -,
: . 1’2014
88
-.
., .,
. ., . . ,
, ,
, .
-,
.
,
.
: (Prunus cerasifera), (Vaccinium myrtilus L),
( Rúmex confértus), (Anethum graveolens), (Prunus domestica), (Petroselinum crispum), (Raphanus sativus).
,
(P. mirabilis, S. aureus MRSA, S. pneumoniae, E. cloaceae, K. pneumoniae, K. oxytoca, P. aeruginosa); (S. enterica, S. flexneri, L. monocytogenes, EPEC E. coli),
(S. mitis, S. mutans, S. pneumoniae)
(E. coli 058, E. faecalis, E. faecium);
(L. acidophilus, L. delbrueckii, L. casei, L. fermentum, B. subtilis 8130, B. subtilis 090, B. dentium). , -
E. faecalis.
P. eruginosa, MRSA, S.
enterica, P. organi . S.
ureus, L. monocytogenes,
S. dysenteriae. -,
E.coli, B.subtilis, E.cloacae.
-.
- -
,
, ,
.
.
., ., ., .
CHLOROBIUM LIMICOLA -8
, ,
,
-. -
: . 1’2014
89
. -
.
- C. limicola -8
.
C. limicola -8. -
C. limicola -8.
0,05 0,5 .
.
GSB.
C. limicola -8
4, 5 6 . ,
,
.
5 -, -
. -
C. limicola -8.
-
. 0,05, 0,125
0,5 - 0,5, 2 3,2
, .
. -
. , -
C. limicola -8
,
.
., ., ., .
S. EPIDERMIDIS
,
, -
,
, -.
, ,
, . --
. ,
-, -
. S.
: . 1’2014
90
epidermidis .
,
-
. ,
-
S. epidermidis 1×106 50 , -
– 1×109 50 . -
,
S. epidermidis,
,
Staphylococcus – 3,5 , --
– 2,3 .
Enterobacteriaceae – 2,5 -
1,2 – -.
-
.
.
. .
-
0,05-5
. , ,
--
Cr( )3. -
( ) 100 1 -
-
-, -
, --
.
, -
. -
h, .
-
.
-
,
100 -
. --
: . 1’2014
91
-
-.
.
. . 1, . . 1, . . 2, . . 2
1
. . , , , ;
2 , , [email protected]
-
,
-, , -
-, . , -
Sphingobacterium multivorum Ochrobactrum antropi
( , -
), , Bacillus
amyloliquefaciens -
.
S 802 (Sphingobacterium multivorum, 109), B 100/13 (Bacillus amyloliquefaciens, 107), S 804 (Ochrobactrum antropi, 109) S 802 + S 804 + B 100/13 1 : 1000.
-
-.
, (2
100 ). – -,
.
10-16% 0,05), -
S 804 -.
12-25% , , -
S 802 .
, -
--
,
, -.
-
, - Sphingobacterium
multivorum, Bacillus amyloliquefaciens Ochrobactrum antropi, -
-.
: . 1’2014
92
., ., ., .
«
», , [email protected]
. , --
,
, . , -
-,
,
.
,
, -
.
,
-, -
. -
, -
:
, -
, -, ,
, -
, -, -
, . --
.
-, -
.
. ,
-
, , ,
-.
., .,
.
-
" " , ,
Candida
-,
: . 1’2014
93
. Candida -
15 % -, 72 %
, 8 % 15 %
-.
.
Candida. -
Candida albicans, -
Candida albicans Candida non-albicans 36% 64%.
-
, C.glabrata(45%), C.tropicalis(35%), C krusei(30%). ,
Candida
2008-2013 . 6,5%.
-
(16,5%) (13,6%).
0,9 %, - 5,4%. -
Candida spp.
, .
9 Candida
C.albicans 74,2%. Candida,
, :
C.albicans, C.tropicalis, C.glabrata, C.parapsilosis, C.sake C.lusitaniae.
C.albicans - 44%,
C.tropicalis 22%. ,
, - C.albicans
94%, C.tropicalis, C.glabrata C.sake
. , Candida
-,
, -
.
., ., .
. , ,
,
. --
. , ,
.
, , ,
, . ,
,
. -
.
,
: . 1’2014
94
.
--
, .
-.
11,4 % 10,2 %
17,5 % 19,6 % , 4,6 5,4
.
, -
, ,
. 16,2 % 18,7 %
21,9 % 23,1 %
6,1 5,9 .
. -
, --
.
., ., ., .
BACILLUS ,
, . , [email protected]
,
Bacillus , -
, -
-, .
,
. -
Bacillus
-, ,
. 16
Bacillus .
1970-77 . --
, - - -
. ,
-
Staph lococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Serratia marcescens, Micrococcus luteus.
: . 1’2014
95
Escerichia coli ATCC 25922 .
--
andida albicans andida tropicalis, andida utilis.
Cryptococcus albidus. -
.
86-98% 10
spergillus sclerotium, Paecilomyces lilacinum, Fusarium culmorium.
Penicillium italicum,
. , ,
andida. 10 -
: .sclerotium, P.lilacinum, F.
culmorium. ,
Bacillus subtilis.
. ., . ,
,
. . . , ,
Cr3+ Cr6+. Cr3+ .
Cr6+ .
,
, . -
Cr6+ Cr3+.
-:
., ,
, . ( 40 ).
5 37 .
Cr6+ Cr3+, -
. - Cr6+ -
-.
, , -, -
-. -
, -
. , -
, . . ,
. , , -
, -. ,
. - 60%,
.
5 ,
.
, -. ,
, ,
.
: . 1’2014
96
, --
. .,
., ., ., ., .
. .
, , , ) . .
. ,
-,
-: ,
, ,
.
Penicillium chrysogenum (60% – 80% - ),
Aspergillus niger (60% 60%), Stachybotrys chartarum (40% 40%), Ulocladium atrum (20% 60%).
-
: – , - –
Acremonium, Aspergillus, Chaetomium, Geotrichum, Penicillium, Stachybotrys, Trichoderma, Ulocladium
gomyc ; – , --
( - ) –
Cladosporium, Alternaria, – -,
– Fusarium,
Botrytys, Phoma i -. -
, : Cladosporium
cladosporioides, C. herbarum, C. sphaerospermum, Scopulariopsis brevicaulis, Ulocladium chartarum,
Trichoderma, , , , :
spergillus ochraceus, A. terreus, A. ustus, A. versicolor, Stachybotrys chartarum.
-,
-. , («
»), , -
.
-
, --
.
., ., ., .,
., ., .
. . , ,
--
.
: . 1’2014
97
, ,
-
. --
( ), -
-. .
. -
« » -
. - -
, - -,
.
-,
. , ,
-
.
-
» « ».
--
. , -
,
OSB- , ,
( -) ,
-, 90%
(29±2)° .
DVD-
. -,
-
, , Ascomycota, -
.
., , ., ,
. .
-
. . , , ;
. , ,
-
- – -
, ,
-.
- -.
--
: . 1’2014
98
,
Fusarium sambucinum – , ,
Mycelia sterilia – , -
-
-. -
2-3 --
, - 40–
42 . -
-, -
, , ,
.
45%, -
70% (4,2--4,4%).
,
, , -.
. -
-, , ,
. -
, -
, – .
-
. -
, .
-, -
-.
. ., . .
PSEUDOMONAS
. , ,
, -.
,
-, , -
.
- Pseudomonas
. -
Pseudomonas chlororaphis (ONU 304, ONU 305, ONU 306), P. fluorescens ONU 303, P. aeruginosa (ATCC 15692, ATCC 27853, ATCC 10145).
:
; , 5 % -; . -
: . 1’2014
99
7 25 37 .
24
. ,
,
Levitch M.E. -
. -,
P. eruginosa ATCC
15692 P. fluorescen ONU 303. , P.
chlororaphis P. aeruginosa ATCC 10145, P. aeruginosa ATCC 27853
. -
-
-.
, --
. -
P. eruginosa - P.
fluorescens P. chlororaphis. , , -
-
.
-.
.
-
. ,
, ,
40 . 10 ,
-, -
.
-
-.
: E.coli,
E.cloacea, K.pneumonia, A.baumanii, P.aeruginos .
. 60
, , - 28(46,66%), -
. ,
E.coli - (55,55%), E.cloacea - (41,66%), K.pneumonia - (38,09%).
-
, - .
P.aeruginosa - (69,23%), , - A.baumanii - (39,63%).
, ,
. 46,66%
: . 1’2014
100
-, 44,89% -
.
-
.
., ., .
, ,
, -
.
, (l gP).
,
( ) -
. - 4- -
- - ( ) -
( )- 46 : 4- -3,5- -4H-1,2,4- ( ), 5- 4- -1,2,4- -3- ( ), 4,5- -4H-1,2,4-
-3- ( ), 5- -1,3,4- -2-
( V), 1- 1- -6- (V),
(VI), 2- -4,6-
)-1,3,5- (VII), 1,2,4- -3- -
(VIII). .
( ) ( ).
-
: ACD Log P: Version 6.0,
Interactive log P calculator (www.molinspiration.com/cgi-bin/properties). ,
(logP -3,52 -1,89),
, .
- VI (logP -0,29 1,28) -
. , VI
.
2- -2-(1,2,3,4--6- )-
(logP -1,99) - - 120
- 21,7±1,7 , -
. (logP -
0,22 2,23), (logP 5,96-7,03), IV (logP 4,00-4,96), V (logP 5,06-6,52), VII (logP 1,32-2,52), VIII (logP -0,20
0,86) .
: . 1’2014
101
. 1, . 1, . 2
ZF40
ERWINIA CAROTOVORA SUBSP. CAROTOVORA
1 . . , ,
2 , ,
[email protected] Erwinia carotovora subsp. carotovora
.
, -
. ZF40 ( -
)
--
. ,
E.
carotovora. --
ZF40. ZF40 ,
– . -
, ZF40
Mu,
E.coli -
, .
-
)
Erwinia carotovora subsp. arotovora
ZF40. 5000 ,
3 , -
( 10-4).
-
, -. -
-
ZF40.
., .
. .
-, ,
, , ,
.
, , -. -
, --
: - ( 64 16 8 )
(
156 ) ,
: . 1’2014
102
. ,
, ,
(800 ), , , -
. -
, , ,
, ,
. -,
, -
.
500
-
-
, --
. -
-.
.1, .1, .2,
.2 .
.2, .1, .1
1 , ;
2 , ,
, .
-,
,
, , ,
.
--
,
5 --
137Cs.
,
. ,
: . 1’2014
103
( ). -
Cladosporium cladosporioides, -
-. ,
-,
.
-
,
.
., .
”
”, ,
. 2012-13 . 732
159 . -
, , -
«bioMerieux» ( )
VITEC2 COMPACT. 159 -,
13,4%.
97,9 %, 2,1% .
(52,9% ) -
S. epidermidis S. aureus.
S.epidermidis 19
14 ( 5 % ). -
. S. aureus
(4 ),
.
Pseudomonas sp., E.coli Acinetobacter baumanii.
.
Candida parapsilosis, 10,2 %
. -
, MRSE – 17,4%, MRSA – 4,4%
. - 4,4
% , 21,7% , --
– 17,4% 47,8 % .
--
.. -
.
: . 1’2014
104
., ., ., .
AZOTOBACTER CHROOCOCCUM
AZOTOBACTER VINELANDII
--
-
.
Azotobacter -
. . chroococcum
. vinelandii, . -
Azotobacter
-. ,
25° ,
, . ,
Azotobacter -
1,25
0,11 , 12,7 .
--
8,4 . ,
1,3
20 40
5,5 1,25 .
1,9–3,9 . -
, A. chroococcum A.
vinelandi , 15 ° .
. -,
40 20 3,7 2,17
, 10,3 10,4
. Azotobacter
4,5–5 .
--
4 3% 10% -
.
.
SACCHAROMY ES CEREVISIAE
..
. , ,
– -
, « »
: . 1’2014
105
(“dancing bodies”). , , -.
-
S. cerevisiae.
S. cerevisiae Y-517 -
( ) . . .
.
-,
DAPI (Sigma, USA). ,
» .
-
.
(40,68 , 30 )
8,2 . 2
« » .
- lag- . -
, « » .
(KCl, KNO3, ) -
,
-
.
S. cerevisiae -
. ,
-.
.
RHODOCOCCUS ERYTHROPOLIS
-,
, -
, ).
. ,
-, -
---
. -
R. erythropolis -.
, -
. , -
, , , -
, -. -
: . 1’2014
106
,
= 8–12 ) ,
, -
( = 4-6 ).
= 1 .
R. erythropolis -23 -
. -
, ( 2,0 ),
(2,0 ) (0,5 )
28,6, 71,4 85,7 %,
,
R. erythropolis -23
.
-
21-52%, 15-76%.
-
.
., ., .
--
. ---
,
. -
,
, ( ), ,
. -
, ,
. ) –
,
, -, -
. ,
( )
,
. --
, -
. - –
-.
.
)
: . 1’2014
107
(Cr, Cu, Pb) .
: 28% (1 ) 37% (5 ) ,
. -
18-26% . ,
, (Cr, Cu,
Pb).
.
. .
( )
.
,
,
, . ,
,
4
). -
-1, -
-.
- ( ),
-.
14 ,
. , -
. Aspergillus, M. racemosus T. viride C. cladosporioides, C. sphaerospermum
S. chartarum, . ,
-1
938 – 7500 ppm, - – 59-
234 ppm. 1875 – 30000
ppm 59-467 ppm, -
. , 1
,
,
. -
1 -, .
: . 1’2014
108
. 1, . 2
1 , . , ;
2 . , ,
-, -
. -
-
, -
. , , -
-
. -
, -,
. -
, , . :
Pseudomonas syringae 8511, Pseudomonas fluorescens 8573, Pectobacterium carotovorum 8982, Xanthomonas campestris pv. campestris 8003 , Clavibacter michiganensis 102, Agrobacterium tumefaciens 8628
, : Dickeya chrysanthemi 8683, Clavibacter michiganensis subsp. sepedonicus 7757, Pectobacterium carotovorum 8982, Pectobacterium carotovorum 8682, Ralstonia
solanacearum 9049 . -
-
12 , -. ,
-
, « ».
+ .,
.
Pseudomonas fluorescens,
. ,
, ,
Clavibacter michiganensis Agrobacterium tumefaciens, , -
.
. ., ., . .
-
. . ,
, , ,
- 26 ) - 20 -
, :
: . 1’2014
109
, , ,
, -,
. -
, , ,
- (12
). , , -
5,67±0,18 lg 2 ,
. -
2,3 ,
2 3 ( < 0,05 ).
, -
-.
.
.
8 , 4
-.
: Moraxella spp., Enterococcus spp., Neisseria spp., Actinomyces spp.
-, ,
. , --
,
-
. -
.
. .
STAPHYLOCOCCUS AUREUS
, ,
-
. -,
, -. , -
, Staphylococcus aureus –
-, , ( -), , , -,
. -
.
1% – ) ( ), -
+370 48-72 .
-, ,
: . 1’2014
110
, , .
,
,
. 48 -
S- -
, 24
. 48
,
.
, 0,6-1,0 , .
Staphylococcus aureus -
.
- 72
+370 , -
.
., .
AZOTOBACTER -
-
. ,
-
,
.
,
( -
, ) ,
(
, ) ,
-.
-
-. -
-
. ,
Roundup. -
Bradyrhizobium japonicum -6035.
--
. 5-6
B. japonicum -6035
- 19 %
( , ).
Azotobacter
: . 1’2014
111
.
7,8 % . ---
: – 13,7
%, – 14,5 %.
. 5-6 -
--
18,3 %,
. -
( -)
Azotobacter 17,5 %.
-:
- – 5,6 %, -
– 8,9 %. , -
, -
, Roundup -
.
: . 1’2014
112
: . 1’2014
113
PLENARY SESSION
Skivka L.M.
IMMUNOGENIC CELL DEATH IN HEALTH AND DISEASES
Taras Shevchenko National University of Kyiv, Kyiv, Ukraine
[email protected] In adult multicellular organisms, homeostasis is maintained through a balance between cell proliferation and cell death. Abnormalities in cell death mechanisms is involved in the patho-genesis of the number of diseases. The failure of cells to undergo apopto-sis participates in the pathogenesis of several human proliferative disorders: malignant transformation, autoimmune diseases, some latent viral infections. A variety of diseases are associated with accelerated rates of cell death: severe -thalassaemia and Parkinson's diseases, polycystic kidney disease and fulminant hepatitis, myocardial infarction and stroke. The immune system is routinely exposed to dead cells during normal cell turnover, injury and infection. The mechanisms our immune system use to deal with dead and dying cells are complex. It is commonly assumed that the immuno-logical consequences of cell death follow a classical dichotomy of immu-nogenic versus non-immunogenic (or even tolerogenic) cell loss. Dying cells release and expose at their surface molecules that signal to the immune system. Potential immunogenic signals emanating from dying cells include proteins that appear at the surface of stressed and dying cells (such as calreticulin and heat shock proteins), lipid moieties that flip from the inner plasma membrane leaflet to the outer leaflet (such as phosphatidylserine), proteins that are released into the
supernatant of cells (such as HMGB1), as well as nucleic acids and their degradation products (oligonucleo-tides, nucleotides, nucleosides, urate) that appear in the extracellular space. Such damage-associated molecular patterns (DAMPs) released from or exposed at the surface of dying cells can drive the pathogenesis of a wide range of inflammatory diseases (acute pancreatitis, gout, chronic obstructive pulmonary disease etc). Main mechan-isms to discriminate between immuno-genic and nonimmunogenic forms of cell death; increasing numbers of endogenous danger signals of the host origin associated with inflammation-related diseases; the use of DAMPs as biomarkers for the diagnosis of inflammatory events in vivo and monitoring of the efficacy of cancer treatment are discussed in the lecture.
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ORAL/POSTER PRESENTATION
Degen A.S., Topol I.O., Kamyshny A.M. FEATURES OF AN EXPRESSION OF
THE T-BET AND GATA3 TRANSCRIPTIONS FACTORS IN EXPERIMENTAL PATHOLOGY
Zaporozhye State Medical University, Zaporozhye, Ukraine
[email protected] Introduction. The great interest represents studying of adaptive immune system, especially tight regulation of the type 1 inflammatory response, at the development of chronic social stress (CSS) and autoimmune disease, such as type 1 diabetes mellitus. The dim of research: To study the peculiarities of T-bet and GATA3 transcriptions factors expression in gut-associated lymphoid tissues (GALT) of rats with experimental STZ induced diabetes mellitus (EDM) and pentoxifilline (PTX) administration and CSS and modulation of the composition of intestinal microflora. Methods: Structure of population of T-bet+ and
: . 1’2014
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GATA +-cells has been studied by the analysis of serial histological sections using the method of indirect immunofluorescense with monoclonal antibodies to T-bet and GATA3 of rat. Results: It has been established that diabetes development was accompanied with 45-72 % ( <0,05) increase in quantity of T-bet +-cells in lymphoid structures of ileum, with 21-44 % ( <0,05) decrease in total density of GATA3+-lymphocytes, insignificant decrease in concentration of T-bet and did not influence on concentration of GATA3 in T-helpers. PTX administration of diabetic animal reduces quantity T-bet+-cells in mucous membrane of villus (on 24-30 %, <0,05), does not influence on density of Th2 in villus, change their number in ILF, reduce concentration of T-bet by the 4th week of development of T1DM at the absence of changes in concentration of GATA3 in immunopositive cell. Development of CSS is associated with an increase of the total number of lymphocytes expressing T-bet in GALT rats with the most pronounced in lymphocyte-filled villi (LFV) (in LFV – on 46%, CSS1 and 92%, CSS2, <0,05). Conclusions: The expression augmentation with T-bet and GATA3 ileum immunopositive cells can influence on differentiation of subsets of T-helpers and their proinflammatory cytokines production, thus acting as one of triggers of diabetes development and progression.
Galkin O.Yu. OPTIMIZATION OF CONDITIONS
FOR THE ISOLATION AND PURIFICATION OF HUMAN IGE
National Technical University of Ukraine "Kyiv Polytechnic Institute", Kyiv, Ukraine
[email protected] To solve many problems in
immunology and molecular medicine are needed in purified preparations of immunoglobulins (Ig). IgE is used as immunogen for immunization and to create immune sorbents designed to remove cross-reacting antibodies and affinity purification of polyclonal anti-Ig sera. Purified IgE is used as standard antigen in the test-kits for IgE quantitative definition and in the tests for anti-IgE antisera evaluation. Following differences between properties of different Ig isotypes and other sera proteins are used for creation of scheme of isolation and purification of human Ig: molecular weight, affinity for proteins A and G, isoelectric point, solubility in different conditions. That is why following methods are used (in various combinations): gel filtration, affinity and ion exchange chromatography, dialysis, precipitation by salts and organic solvents. The aim of the study was to develop improved methods of isolation and purification of human IgE, suitable for use in highly sensitive immunoassay methods. Based on the available bibliographic data and our own experience it has been theoretically synthesized and experimentally tested the following scheme of isolation and purification of human IgE: 1) removal of serum human IgG using affinity chromatography on protein G; 2) removal of serum human IgA and IgM using immunoaffinity sorbents based on anti-IgA and anti-IgM monoclonal antibodies that were at our disposal; 3) the allocation of human IgE in comparative experiments using sephacryl S-300 and superdex 200; 4) control the quality of the IgE antibodies by means of Ouchterlony double immunodiffusion and electrophoresis
: . 1’2014
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in polyacrylamide gel (reducing conditions). Using the proposed scheme has allowed to obtain high purity human IgE fraction. Obtained human IgE is suitable to be used for immunization and highly sensitive methods of immune analysis. Yield of human IgE after all phases of purification has been about 42% of the initial amount of immunoglobulin E in serum.
Kiyamova R. G.1 Kostianets .I.1 Dyachenko L.V.2, Lytovchenko A.S.2,
Filonenko V.V1. TUMOR-ASSOCIATED ANTIGENS AS MOLECULAR MARKERS FOR BREAST CANCER DIAGNOSTICS
1 Institute of Molecular Biology and Genetics, NAS of Ukraine, Kyiv, Ukraine
2 Taras Shevchenko National University of Kyiv, Kyiv, Ukraine
[email protected] Breast cancer is the most widespread cancer type in women. Many breast tumors with high histological tumor grade, mitotic index and proliferation rate which overrepresented among the so-called interval breast cancers (eg, cancers arising between annual mammograms) cannot be detected by currently existing methods, in particular mammography. In a view of above mentioned facts an urgent need exists for creation and development of new alternative methods to improve breast cancer diagnostics and prognosis. Over the last few years emerging evidence suggests that tumor-associated antigens (TAAs) and their cognate autoantibodies serve as molecular markers of human malignancy. The principle of the immunological detection of molecular changes in tumor cells by autoantibodies allowed us to identify 41 autoantigens from medullary breast
carcinoma (MBC) tumors by SEREX (serological analysis of recombinant tumor cDNA expression libraries) approach. Large-scale allogenic screening of breast cancer antigens with sera of breast cancer patients of different histological types and sera of healthy individuals allowed us to reveal that 6 TAAs including RAD50, PARD3, SPP1, SAP30BP, NY-BR-62 and NY-CO-58 had higher immunogenicity in sera of breast cancer patients compare with sera of healthy individuals. Combination of these 6 TAAs in a single panel with Roc-analysis may differentiate cancer patients and healthy individuals with 70% of sensitivity and 91% specificity. This panel of 6 TAAs could be considered as the base for creating of serological test-system for non-invasive breast cancer diagnostics in future. The sensitivity of proposed panel can be increased by extending of TAA panel. Analysis of immunogenicity of additional TAA identified by SEREX screening in sera of breast cancer patients is currently in progress.
Nikulina V., Garmanchuk L., Senchylo N., Nikolaienko T.
ADHESION POTENTIAL, PROLIFERATION AND THE
GLUCOSE ABSORPTION LEVEL OF THE HELA CELLS UNDER THE
INFLUENCE OF TEICHOIC ACID Taras Shevchenko National University of
Kyiv, Kyiv, Ukraine [email protected]
The modern researches reveal anticarcinogenic influence one of major bacteria cell wall components – teichoic acids (TA). It was shown that TA of cell wall of some bacterium is responsible for enhancement of hypersensitivity reaction and able to
: . 1’2014
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activate cell cytotoxicity, supreses antibody synthesis in big concentrations. Many researchers underline particular role of toll-like receptors (TLRs) which recognize different bacterial structures and induct antitumor effect. This receptors are expressed on surface of different cell types including different types of tumors. The interaction TA and LTA with specific receptors results in activation of links of antitumor immunity. Our previous studies on transplantable Lewis lung carcinoma have revealed antitumor and antimetastatic effects of the bimetallic complex of copper and cadmium in a combined application with teichoic acid obtained from St. aureus Wood 46, but study of application of TA alone was not conducted. The aim of our study was the exploration of the TA influence on an adhesion potential, proliferation and the glucose absorption level of the Hela cells under standard culture conditions. The change of adhesion potential of HeLa cells was detected. This parameter was about 10% lower on exponential phase of growth and 20% higher on stationary phase. The glucose absorption level was lower by 30% in presence of TA on exponential phase of growth and 70% higher on stationary phase. The changes in proliferation rate were shown under the influence of TA, proliferation on exponential phase was increased by 40%, when proliferation on stationary phase was decreased by 35% in comparison with control, according to data of counting in presence of trypan blue. In conclusion, earned data indicated the rise of malignant degree of tumor cells after the influence of the teichoic acid. Also, in our previous
studies we have revealed that TA in combined application is capable of altering the adhesive characteristics of tumor cells, which may be caused by its effects on TLRs. Definition of antitumor efficacy of TA needs further investigations in combination with classic treatment agents.
Palyvoda K.O., Oliinyk O.S.,
Lugovskaya N.E., Kolibo D.V., Lugovskoy E.V., Komisarenko S.V.
GENERATION AND CHARACTERISATION OF
RECOMBINANT SINGLE CHAIN VARIABLE FRAGMENT ANTIBODIES
AGAINST PRO186-LEU197 PROTEIN C REGION
Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine,
Kyiv, Ukraine [email protected]
Protein C also known as blood coagulation factor XIV is a vitamin K-dependent serine protease which is a major component of human blood anticoagulation system. Anti-protein C antibodies have been reported as a tool for protein C level measure in human plasma under normal and pathological conditions. For this purpose single-chain fragment variable (scFv) antibodies are preferred than traditional monoclonal antibodies due to their greatly reduced size and ease of genetic manipulation. The aim of this research was to obtain and characterise recombinant scFv antibodies specific to Pro186-Leu197 protein C region. Pro186-Leu197 protein C region was chosen because of its low homology to other plasma proteins and high immunogenicity. Pro186-Leu197 peptide was synthetized following the methodology of peptide synthesis in solid phase and conjugated to keyhole limpet
: . 1’2014
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hemocyanin (KLH) or bovine serum albumin (BSA). A phage display immune library of immunized with Pro186-Leu197 protein C region-KLH conjugate mice was generated. The library was screened against Pro186-Leu197 protein C region-KLH and -BSA conjugates and one scFv producer clone was isolated. The results showed that obtained scFv antibodies could specific bind both Pro186-Leu197 protein C region-BSA conjugate and activated protein C. ScFv recognized target antigen in Western blotting, but only under non-reducing conditions. Isolated clone DNA sequence was subcloned into expression vector for easily scFv isolation. Thus, in the present study scFv antibodies against Pro186-Leu197 protein C region were obtained. Our data indicate that these scFv are suitable for ELISA, Western blot analysis and immuncytochemistry and may be applied in protein C research and possibly for new diagnostic systems design.
Siryk G.1, Fedorchuk O.2, Malanchuk O.3, Skivka L. 1
PHYSICAL ACTIVITY AT DIFFERENT DAYPARTS CHANGES
CIRCULATING PHAGOCYTES FUNCTIONS
1Taras Shevchenco National University of Kyiv, Kyiv, Ukrain
2R. E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology,
Kyiv, Ukrain 3Institute of Molecular Biology and
Genetics, Kyiv, Ukrain [email protected]
An ever-growing volume of peer-reviewed publications speaks to the recent and rapid growth in both scope and understanding of exercise immunology. Regular exercise and
physical activity provide many health benefits and are encouraged by medical professionals for the prevention of illnesses. Nevertheless, it is known that exercise can have both positive and negative effects on immune function, depending on the intensity and duration of physical activity. Immune cells have diverse responses to acute exercise or long-term training at moderate and high intensities. The present study focuses on the effects of moderate exercises performed in different day parts on circulating phagocytes functions in healthy adult men. Six healthy volunteers were recruited; blood samples were obtained before and after standardized bout of moderate physical activity which was performed at noon and at midnight. ROS (reactive oxygen species) generation and phagocytosis were estimated by flow cytometry. The reaction of circulating phagocytes on the bout of exercises which was performed in the day time differed from that on exercises which was performed at night. We also registered some differences in the reactions of monocytes and neutrophils on moderate exercises. Phagocytic activity of monocytes and neutrophils had increased after physical activity which was performed in the day time and at night. Standartized bout of moderate exercises which was performed in the day time caused the decrease of ROS generation by both monocytes and neutrophils whereas physical activity at night was associated with the increase of oxidative metabolism of circulating phagocytes.
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; - -
Cu 6,3%; 4,2% 6,9%
: . 1’2014
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: . 1’2014
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-
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. -
CD8 NK 654 IVF .
CD8 NK 90 -
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(>60%) (OR 2.85 p=0.053) (<40%)
(OR 3.565 p=0.018). ,
– CD8 NK
(OR 5.6 p=0.0007). -
N CD8 -
CD8 N CD8 . 654
CD8 N
13% 86%.
N IR CD158a
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: . 1’2014
142
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: . 1’2014
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, 1 .
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/ , - --
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: . 1’2014
144
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– 650 , 530 10- .
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-,
. -
: . 1’2014
146
-
, . -
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. 96-
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, 9,2). -
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., ., ., , ., .
-
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. . - HBc,
. ,
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,
. -, -
(IgM IgG) HBc. -
: . 1’2014
147
, -
. HBcore -
E. coli BL21(DE3)CodonPlus-RIL, 24
HBcAg.
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HBcore -.
-
- 95 %. -
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. - HBcore
, ---
, .
HBcore, . -
,
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HBcore,
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2 -
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. , .
. :
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e
. per os (500 25 )
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(20- )
, 500
, , .
: . 1’2014
148
– ,
( ). -
, 500
,
, .
25 (
)
, (80% 65%
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(6,4±2,7/ 13,9±4,5/ – ).
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., .
XXI ! -
, 2-3 10
, .
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. 2008
14 910 600 -. -
,
: 10 30 %
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45% - 8
; 40 % -
8 -;
--
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, -
: . 1’2014
149
.
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. , -
. -
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, , , ,
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. -
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: - : 1. -
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- 63,2%; 4. -
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. : -
: 1. - – 35,5%; 2.
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- 27%; 5. –
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1 3- 0,005 2-3 ; - 3 7 - 0,01 2
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10-15 . .
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. : -
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,
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: . 1’2014
150
--
,
-. III.
, , -
, . -
– . : 2,4,6,8– –
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. : – 300 , 500 . -
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: . 1’2014
151
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,
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VITRO IN VIVO «
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( Fe) --
, -
, , . ,
Fe ,
.
Fe in vitro in vivo . -
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. . . Fe
IgG, -
in vitro;
, 1 24
- Fe.
, in vitro Fe
1 ,
Fe 24
. IgG Fe 0,1; 0,01; 0,001
, ,
35, 15 1 . Fe 0,5
86,9 % . 1
Fe -
: . 1’2014
152
( 6,5% 17,2%).
24 ( 25,5%
22,8%). Fe -
, -
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1 24 Fe
-
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-,
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in vitro ,
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., ., ., .,
., .
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, ,
,
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,
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.
--
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,
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-
: . 1’2014
153
, .
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.
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« . . .
», , [email protected]
, , ( ) -
-
: . 1’2014
154
-
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,
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( )
.
: 1- 3-
3- , 21- . - . 1-
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- 6-
0,94±0,05 , 10-
0,78 ±0,05 , 14- 0,16±0,08 .
3-
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0,93±0,01 , 14- 0,66±0,06 ,
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», , [email protected]
-
: . 1’2014
155
. ,
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: ( 6 ) — -
; ( 7-9 ) —
: . 1’2014
156
;
10 ) — , -,
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. ,
(1,076-1,077 ) (1,087 ), , -
( ) . , -
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. -:
( , n=10) (
, n=10). -
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[Q1; Q3] — 43,0 [31,5; 59,8] %), ( 8,7-10,8 , 45,0 [31,8; 57,0] %)
( 11,1 , 10,0 [7,5; 12,3] %). -
-
. -
( <0,05, )
(7 [6; 8] %) ,
- (39,0 [36,5; 40,0] %).
(54,0 [53,8; 56,5] %) (p>0,05)
. -
-.
.
- -4 ,
. .
-,
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-
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122 - ( ) -I. 40
( ) -1 .
, -
28,6±1,2
,
: . 1’2014
157
24,5±13,3 , 2
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., ., ., .,
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10 65 %. 30 % -
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-
. 9
15 ), -: - ,
; - ,
.
, sIgA
,
-.
sIgA
-. , ,
, , -
, ( sIgA
2 ). ,
,
,
: . 1’2014
158
-
, 40% -
, -. ,
sIgA .
.
., ., .
-123
« ,
», , [email protected]
) – ,
- ,
.
,
. -.
--
.
- 123 - ("Burst
test"). -,
-
-123 - - -
-123). --
(fMLP), - E.coli,
– – ( ). –
.
.
R848( ) –
7/8, -, ). -
-
-). ,
85,6% 82,6%, .
- E.coli - 98,7%.
, -
-
20,6% - E.coli, 12,1% 17,8%
. R848
.
,
, --
.
: . 1’2014
159
.
-
-
. .
NO,
, .
NO ,
-
(LAR).
: 1
(100 (OVA) 100 Al(OH)3) 5
(100 OVA 10 Al(OH)3).
0,1% OVA 0,9% NaCl -
OVA sIgG ELISA
. NO -
( ) -
(Beckman Coulter — EPICS C)
DAF-2DA. -
0,2% Trypan Blue Neubauer. , OVA sIgG -
12 — 20 -
.
OVA sIgG - 2
, OVA,
- 0,9% NaCl. -
, DAF-2DA -
OVA 2,6
DAF-2DA
0,9% NaCl. ,
-,
,
, -
, - NO -
LAR
0,9% NaCl, OVA
sIgG .
.
80 « »
, ,
1933 . -,
, , 1933
1951 -.
-
: . 1’2014
160
. 1951
.
1953
– -
.
1000 , 6
-.
(1970-1971) -
-
. 1972 1974 -,
-
. 1983 . .
.
-
-. 1986
-
-
-
.
, --
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-
-, .
. «
» --
.
., ., ., .,
.
,
», ,
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--
-
. --
, --
VEGF .
, VEGF
,
( / 25 :
: . 1’2014
161
75 ) 2646 / 2162 : 2756 .
, (rP = 0726)
VEGF –
” ( = 0.027).
: Var1 = 20.46*Var4 – 75.93, Var1 – -
VEGF , Var4 – –
“ ”. VEGF -
1 -
( >0,05), -
>0,05), . VEGF
( >0,05).
VEGF
>0,05), (p= 0,165),
(p= 0,543), (p=0,472).
VEGF -
, -. , -
,
VEGF -.
- VEGF .
.1, .2
1
, , 2
---,
,
.
, -
. -,
, -, ,
,
. -
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-,
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: . 1’2014
162
,
- .
, -, ,
.
, -
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-
,
.
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,
, ,
–
.
1986 .
-137 -90, ,
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-
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, ,
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:
,
-, 1995
2013 . - 5 .
,
-
, -
, -, .
, -
, -,
1995 2013 , .
. -
CD3+ CD4+,
CD4+/CD8+ -
. , -
. ,
: . 1’2014
163
-. -
-, ,
, -
.
., ., ., .
», ,
. ,
-.
- ,
. .
74 -
15 82 , – 21-
, 13 – 40 – . -
52 . -1 , 2, 4, 6,
10, 13, -a -
« », « », «Diaclone».
Statistika 6,0.
,
-
,
.
, .
: 1. -,
-
. 2. , ,
. 3. --
.
., ., ., .
, - Mollicutes
-, -
. , , -
, ,
, ., -
-.
-
: . 1’2014
164
, -
, -,
.
-,
. , -
-. ,
Acholeplasmataceae. , -
, ,
, --
. --
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laidlawii – oculi” --
, -
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.
., .
CD45RA/CD45ROCD4+ ,
«
. . », ,
. , ,
-.
CD45 CD4+ -
-.
CD4+CD45RA-CD45R0+
CD4+CD45RA+CD45R0- -
. -
CD4+CD45RA-CD45R0+
in vitro.
CD45RA CD45ROCD4+ -
.
.
: . 1’2014
165
: - 10-16 , - 30-85 .
-.
CD45RA CD45ROCD4+
.
,
, .
(>1:32) : 42,9% - 1,2 , 14,3% - 3
; – 33,3% , 8,3% - 2,3 .
14 . -
CD45RA CD45ROCD4+
: 33,67%-CD4+CD45RA-
CD45R0+ , 61,49% - CD4+CD45RA+CD45R0- .
-: 67,46% - CD4+ -
, 27,1% - CD4+ . : 1)
; 2)
-
CD45RA CD45ROCD4+ -
.
., ., ., .
, ,
( ) , ,
( )
( ), ,
, .
140 -
. -
, , , --
. -
CD4- CD8
CD16- .
( -1 , -8, - ).
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-: (Hirudo verbana,
Carena, 1820), (H. medicinalis, Linnaeus, 1758) (H. rientalis, S. Utevsky et Trontelj, 2005) ,
, . -
, -.
, -
-,
-
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: . 1’2014
166
-
: -, , -
, , ,
. 5 : - (11,8 %
), , ( -
, 85,7; 2,0 0,3 % ), - –
(0,2 % ). -
.
" », ,
-, -
. 90 % -
. -
: Esherichia.coli
,
, , Staphylococcus
aureus, Streptococcus agalactiae
. , -
.
. 20
.
- (Himedia,
). -
"Lachema" ( ). -
RIDASCREEN FAST Lysozym Biopharm ( ). -
33 , S. aureus
(n=5), Staphylococcus spp. (n=10), S. agalactiae (n=12), Enterococcus spp. (n=6).
-.
, -
( 0,02 ) -
. 3- ,
-. , ,
,
.
-
.
.
: . 1’2014
167
., .
,
,
.
-
. -
. (SVC) -
, Rhabdoviridae
Rhabdovirus.
SVC -
, ( )
( ).
,
(32,3% <0,001). -
- 30,3%.
.
. -
, -.
. ( )
.
-.
, ,
.
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. ,
, . -
,
, .
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1986-87 .
1 , ,
2 “ ”,
,
,
.
: . 1’2014
168
.
( ) 137 ( ) .
4 (D)
: - (0<d<d<d<d2 cSv, . -
, --
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. -
. -
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2 Sv. -.
CD3+HLA-DR+
, - 10 Sv. .
,
-
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., .
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-
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.
: . 1’2014
169
.
Fomes
fomentarius »,
2002 . , ,
. « » -
, ,
.
,
.
.2 .2, .1 .2,
.1 .1
PSEUDOMONAS AERUGINOSA
ARABIDOPSIS THALIANA
1
, , 2
( )
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, --
. -, -
-
, -
.
( 0,85 %
) Pseudomonas
aeruginosa - -8614 -9096
Arabidopsis
thaliana (Col-0 wt) , -
(Jin) (Nah G) .
100 .
P. syringae -8511.
8614 (145–
275 %), - ( 26–69
% Nah G 62–88 % Jin).
8614 .
9096
. - 9096
Col-0 wt,
. ,
: . 1’2014
170
, , .
., .,
., ., ., ., .,
. « »,
,
-, -
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1 ).
2
( -), -
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: . 1’2014
171
., ., .
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[email protected] , 13,3 .
15 .
. ,
, 36 000 86
000 – . 2013 18735 -
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:
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-- 3860 ; -
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104 (22,6%); – 126 (27,3%); -
33 (7,2%). 3. 7424 (
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; : - 10
(33,3%); - - 8 (26,7%); -
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: . 1’2014
172
. ,
, -
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-, .
4 : ,
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3 52% -
, -, -
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3 (p<0,05).
, , -
2 (p<0,05). : -
.
: . 1’2014
173
: . 1’2014
174
ORAL/POSTER PRESENTATION
. . «
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: . 1’2014
175
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