Vol 93, No 6 (2016)

ORIGINAL RESEARCHES

FEATURES OF FORMATION OF BACTERIAL BIOFILMS IN CONDITIONS OF SPACE FLIGHT

Rybalchenko O.V., Orlova O.G., Vishnevskaya O.N., Kapustina V.V., Potokin I.L., Lavrikova V.V.

Abstract

Aim. Study the effect of microgravitation on the formation of Lactobacillus plantarum 8PA-3 bacterial biofilms in the conditions of space flight. Materials and methods. Information on the effect of microgravitation on the development of biofilms was obtained during study of L. plantarum 8PA-3 probiotic lactobacilli in special equipment in the process of execution of space experiments in the Russian segment of the International Space Station. Comparative analysis of growth of plankton and biofilm forms of cells developing in the conditions of space flight and surface conditions was carried out by microbiologic and electron-microscopy methods using scanning and transmission electron microscopy. Results. Accelerated dynamics of formation of L. plantarum 8PA-3 lactobacilli biofilm on the surface of polymer substrate was shown for the first time during the space experiment. Microbiological analysis of the bacterial culture has also confirmed the accelerated growth of L. plantarum 8PA-3 under microgravitation compared with surface conditions. Ultrastructure of plankton form of L. plantarum 8PA-3 taking part in formation of biofilms in conditions of microgravitation was detected for the first time in the space experiment. Conclusion. Data on comparative electron-microscopic analysis obtained in space experiments are important for scientific justification of the effect of microgravitation on bacterial communities developing as biofilms - the most natural form of existence of microorganisms. The results obtained could be taken into consideration during creation of novel antibacterial means and disinfectants as well as methods of treatment of surfaces of modules of piloted space complexes that could allow to clarify methods of effective prophylaxis of biofilm spread which pose a risk of health of the crew and normal functioning of equipment in the International Space Station.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):3-10
pages 3-10 views

BIOLOGICAL ACTIVITY OF YERSINIA PSEUDOTUBERCULOSIS TOXINS

Terentieva N.A., Timchenko N.F., Golotin V.A., Rasskazov V.A.

Abstract

Aim. Study of effect of heat-labile (HLT) and thermostable (HST) lethal toxins of Yersinia pseudotuberculosis on the development of embryos of sea urchin Strongylocentrotus intermedius, processes of biosynthesis of nucleic acids and protein in embryo cells and activity of nucleoside-kinases of sea urchin. Materials and methods. Y. pseudotuberculosis strains 2517 (pYV-) and 512 (pYV48MD, pYV82MD) were used for isolation of HLT and HST. Gametes and embryos of sea urchin S. intermedius were used to carry out the experiments and isolate nucleoside-kinases. Results. Both of the studied toxins of Y. pseudotuberculosis possessed spermiotoxic effect and reduced fertilizing ability of sea urchin spermies. HLT LD50 was 1 (ig/ml, and HST - 2 pg/ml. Toxins affected the development of embryos of sea urchin resulting in severe morphologic damages, cessation of the development of embryos at early stages of embryogenesis, destruction of cells and death of embryos. Wherein, damaging effect of HLT was observed at lower concentrations compared with HST. HLT inhibited DNA and RNA biosynthesis at concentrations of 1-2 pg/ml. HST did not affect biosynthesis of nucleic acids even at high concentrations, but inhibited protein biosynthesis in sea urchin embryos. HLT did not reduce the level of inclusion of labeled amino acids into embryo cells. HLT had inhibiting effect on the activity of thymidine- and uridine-kinase of sea urchin, whereas HST did not affect these enzymes. Conclusion. Both of Y. pseudotuberculosis protein toxins affect the development of sea urchin embryos, however, mechanisms of action of HLT and HST on embryos and processes occurring in them differ.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):10-19
pages 10-19 views

GIS: CAPABILITIES OF DATA ANALYSIS OF PHENO- AND GENOTYPING OF EL TOR 01 SEROGROUP CHOLERA VIBRIOS ISOLATED FROM AQUATIC OBJECTS OF THE ENVIRONMENT IN RUSSIA FEDERATION

Levchenko D.A., Kruglikov V.D., Vodopianov A.S., Titova S.V., Arkhangelskaya I.V., Nepomnyaschaya N.B., Ezhova M.I.

Abstract

Aim. Application of the authors’ GIS «Cholera 1989-2014» for systematization of atoxigenic strains of serogroup 01 cholera vibrios (ctxAB-tcpA-, ctxAB-tcpA+), isolated from aquatic objects of the environment by pheno- and genotype. Materials and methods. A sample of 304 Vibrio cholerae 01 strains was studied. Isolation of 39 genes related to pathogenicity was carried out. Discrimination ability of a set of genes was determined by Simpson formula. Cluster analysis was carried out by UPGMA method. Results. Analysis of multi-year data on aquatic V. cholerae 01 strains in country’s subject was carried out using GIS. A possibility of systematization of phenotypes of the isolated strains by defined parameters was shown. An experimental program for detection of presence/lack of various genes and their combinations forgenotyping was developed. Conclusion. GIS was established to allow to carry out analysis of phenotypes by defined parameters, as well as implement approximate systematization of genotypes of atoxigenic strains of cholera vibrios 01 by optimally sufficient detection of 14 genes.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):19-25
pages 19-25 views

IDENTIFICATION OF CAUSATIVE AGENTS OF GLANDERS AND MELIOIDOSIS BASED ON PRINCIPLES OF POLYPHASE TAXONOMIC APPROACH

Korsakova I.I., Antonov V.A., Khrapova N.P., Zamarina T.V., Pimenova E.V., Kim E.E., Merinova L.K., Senina T.V., Tkachenko G.A., Savchenko S.S., Ageeva N.P., Molchanova E.V., Lopasteiskaya Y.A., Prokhvatilova E.V.

Abstract

Aim. Determine an optimal set of the most effective methods of identification and intraspecies typing of causative agents of glanders and melioidosis. Materials and methods. Bacteriologic, immunochemical, molecular-genetic methods were used. Results. A possibility to identify collection strains of pathogenic and closely related Burkholderia in semiautomatic systems is studied. Means of detection of informative variable genome segments of the specified microorganisms were developed, methods of their genetic typing were selected. Effectiveness of application of precipitating mAbs for differentiation of Burkholderia was established. Data on diagnostic possibilities of immunoglobulins fluorescing based on monoclonal antibodies of various etiotropic directionality for detection and identification of B. mallei and B. pseudomallei are generalized. Experimental series of amplification test-systems for identification of glanders and melioidosis causative agents in real-time PCR format are created. Conclusion. A number of methods for identification and typing of glanders and melioidosis causative agents is proposed.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):25-34
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POPULATION MORBIDITY BY INFECTIONS TRANSMITTED VIA IXODES PER-SULCATUS IN THE NORTH AND SOUTH OF IRKUTSK REGION

Nikitin A.Y., Noskov A.K., Balandina T.P.

Abstract

Aim. Evaluate the degree of epidemic risk of emergence of tick-borne encephalitis (TBE) and ixodes ticks’ borreliosis (ITB) in municipalities (MPs) of Irkutsk region with various natural-climate conditions. Materials and methods. Morbidity was compared for TBE and ITB during 2001 - 2015 in MPs of Irkutsk region located to the north or south of the 55th parallel, i.e. in the conditions of different severity of sharply continental climate. 5-year average data were analyzed. Results. ITB morbidity was 2 - 3 times higher than TBE for all the 5-year periods in the north of the region, whereas in the south -differences were not present. Moreover, in MPs located to the north of the 55th parallel in 2001 - 2015 a decrease of TBE and ITB morbidity did not occur, as in the south of the region. Conclusion. The lack of population morbidity reduction by TBE and ITB in the north and prevalence of the latter nosoform requires enhancement of a complex of prophylaxis measures in these MPs with an accent in development of non-specific means, including acaricidic treatment, enhancement of operations of centers of express diagnostics of transmissible infections, familiarizing of the population with the use of protective suits during periods of high activity of ticks.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):34-40
pages 34-40 views

GUT MICROSYMBIOCENOSIS IN CHILDREN WITH REACTIVE ARTHRITIS

Bukharin O.V., Chelpachenko O.E., Danilova E.I., Chainikova I.N., Perunova N.B., Ivanova E.V., Nikiforov I.A., Fedotova L.P., Bondarenko T.A., Salgina A.V.

Abstract

Aim. To study the state of gut microsymbiocenosis in children with reactive arthritis (RA), with the assessment of biofilm formation (BFF) of microsymbionts and the ability to change cytokine levels (their anticytokine activity) in vitro. Materials and methods. The investigation of gut microsymbiocenosis by means of bacteriological method was conducted in 34 children with RA and 25 relatively healthy 3 - 16 year- old children. Microorganisms were identified with the help of MALDI-TOF mass-spectrometry, anticytokine activity (АСА) of microsymbionts - according to Bukharin O.V. et al. (2011), biofilm formation - according to O’Toole G.A., Kolter R. (1998). Results. On the ground of species composition differences of gut microbiota discrimination model was created which allowed to separate the group of children with RA from healthy individuals. Microsymbiocenosis of patients with RAwas characterized by increasing number of opportunistic microorganisms (OM) (enterobacteria, clostridia, bacteroides, and Candida), BFF and АСА level. Conclusion. The obtained data greatly contribute to the deciphering of spondylo-arthritis and disclose the role of microbial factor under given pathology. Hypercolonisation of human gut with OM, having pronounced ability to BFF and regulating cytokine level, promotes strengthening of arthritogenic potential and serves as additional marker of arthritis development risk in children.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):41-48
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PROFILES OF CYTOKINES IN MICE DURING IMMUNIZATION WITH ADTP-VACCINE WITH ACELLULAR PERTUSSIS COMPONENT

Zaitsev E.M., Poddubikov A.V., Britsina M.V., Ozeretskovskaya M.N., Mertsalova N.U., Bazhanova I.G.

Abstract

Aim. Study cytokine status in mice immunized with vaccines containing acellular pertussis component. Materials and methods. Vaccines developed in Mechnikov RIVS - acellular pertussis vaccine (aPV) and adsorbed pertussis-diphtheria-tetanus vaccine (aDTaP), containing a complex of protective antigens of pertussis microbe - were used in the study. Fi (CBAxCsyBle) line mice weighing 12 - 14 g were immunized intraperitoneally 3 times at an interval of 7 days with aPV and aDTaP at human immunization dose (0.5 ml), containing 25 pg of pertussis component. Intact mice were used as a control group. Levels of IFN-y, IL-2, IL-4, IL-5, IL-12 cytokines were determined after each immunization in enzyme immunoassay using commercial test-systems from Cusabio (China). Results. An increase of levels of IFN-y, IL-2, IL-5, IL-12 and lack of stimulation of production of IL-4 was established in dynamics of immune response after administration of aPV and aDTaP vaccines. Conclusion. The data obtained indicate that immunization of mice with aPV and aDTaP vaccines resulted in activation of production of cytokines characteristic for immune response during pertussis infection and immunization with whole-cellular aDTP-vaccines.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):49-53
pages 49-53 views

SYNTHETIC CONJUGATED ANALOGUES OF CAPSULE POLYSACCHARIDES OF PNEUMOCOCCUS - AN INSTRUMENT FOR DETECTION OF POST-VACCINATION ANTIBODIES

Kurbatova E.A., Akhmatova E.A., Akhmatova N.K., Egorova N.B., Yastrebova N.E., Sukhova E.V., Tsvetkov Y.E., Yashunsky D.V., Nifantiev N.E.

Abstract

Aim. Evaluation of the ability of capsule polysaccharides (CP) of Streptococcus pneumoniae serotype 3 and 14 and their synthetic structure analogues, conjugated with bovine serum albumin (BSA), to detect antibodies in post-vaccination sera of mice. Materials and methods. Oligosaccharides corresponding to one, one and a half and two repeating links of serotype 3 and 14 S. pneumoniae CP were synthesized, their conjugates with BSA were produced by squarate method as well. Ligand content per BSA molecule was controlled by MALDI-TOF spectrometry. Immune sera were obtained after 2 intraperi-toneal administrations to mice of glucoconjugates adsorbed on aluminum hydroxide or 13-valent pneumococcal conjugated vaccine. Determination of levels of post-vaccination class G antibodies and their sub-isotypes was carried out in EIA. Results. Immunization of mice with neoglucoconjugates resulted in formation of predominantly IgGl recognizing serotype 3 and 14 S. pneumoniae CP. IgGl in mice immunized with a 13-valent conjugated vaccine recognized serotype 3 S. pneumoniae CP, but detected serotype 14 S. pneumoniae CP weakly. All the conjugated synthetic oligosaccharides were characterized by a high ability to bind antibodies in blood of mice immunized with the polysaccharide conjugated vaccine. BSA-tetrasaccharide of serotype 3 S. pneumoniae and BSA-tetrasaccharide of serotype 14 S. pneumoniae were characterized by the highest ability to detect IgG 1 against CP. Conclusion. Synthetic oligosaccharides, conjugated with BSA protein-carrier, may be used to develop diagnostic test-systems for determination of antibodies in post-vaccination sera.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):54-60
pages 54-60 views

COMPARATIVE ACTIVITY OF INFLUENZA VACCINES: EFFECT ON LYMPHOCYTE SUBPOPULATION STRUCTURE

Chromova E.A., Semochkin L.A., Akhmatova E.A., Stolpnikova V.N., Skhodova S.A., Sorokina E.V., Akhmatova N.K., Kostinov M.P.

Abstract

Aim. Study subpopulation structure of lymphocytes in healthy individuals under the effect of various influenza vaccines in an in vitro system. Materials and methods. Evaluation of immunephenotype features of PBMCs, activated in vitro by immune-adjuvanted and unadjuvanted vaccines against influenza in healthy individuals, was carried out by using flow cytometry method. Results. Grippol plus vaccine caused a more pronounced stimulating effect compared with subunit and split-vaccines on NK-cells, cells with markers of early activation CD45/CD25, induced the quantity of natural regulatory cells (CD4/CD25/Foxp3), increase of the number of В-cells and reduced the amount of cell types with apoptosis marker CD45/CD95. Conclusion. Immune-adjuvanted vaccine Grippol plus induced formation of effectors of both innate and adaptive immunity and possessed the most powerful potential regarding activation of various types of immune-competent cells compared with unadjuvanted vaccines.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):61-65
pages 61-65 views

EXPERIMENTAL RATIONALE OF ENDODONTIC THERAPY OF CHRONIC FORMS OF PULPITIS AND PERIODONTITIS USING PHOTO-ACTIVATED DISINFECTION AND ULTRASOUND TREATMENT

Tsarev V.N., Podporin M.S., Ippolitov E.V., Avtandilov G.A., Tsareva T.V.

Abstract

Aim. Laboratory rationale of application of a complex effect of ultrasound treatment and photo-activated disinfection in experiments in vitro using strains of anaerobic and microaerophilic microorganisms isolated from patients with chronic forms of pulpitis and periodontitis. Materials and methods. Microbiologic study of root canal was carried out using a bacteriologic method (2 species of obligatory-anaerobic and 2 - microaerophilic streptococci). Evaluation of the effect of ultrasound treatment in a complex with a system of photo-activated disinfection on microflora of root canal was carried out using a bioreactor. Study of biofilms of the root canal system of a tooth in patients with chronic forms of pulpitis (52 individuals) and periodontitis (16 individuals) were carried out using scanning electron microscopy. Results. The presence of an ample microbial biofilm in the ostiums of dentinal canaliculi and obturation of dentinal canaliculi in certain cases was established. Features of formation of growth curves for bacterial populations of oral microflora (Porphyromonas gingivalis, Fusobacterium nucleatum, Streptococcus sanguinis, Streptococcus mutans) were described in the experimental part of the study as a result of a complex effect of photodynamic activation and ultrasound compared with control lacking those. Conclusion. The data obtained allow to conclude that complex effect significantly suppresses the growth or microbial populations of oral bacteria.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):66-73
pages 66-73 views

REVIEWS

PLASMID-ASSOCIATED VIRULENCE OF YERSINIA PSEUDOTUBERCULOSIS AND INFECTIOUS PROCESS

Somova L.M., Shubin F.N., Drobot E.I., Plekhova N.G., Lyapun I.N.

Abstract

Literature data regarding genetically determined pathogenicity factors of Y. pseudotuberculosis and associated manifestations of this infection caused by various plasmid types of the causative agent are generalized. Principal attention is given to features of cell-tissue alterations mediated by virulence plasmid pYV, as well as effects of pathogenicity of an understudied pVM82 plasmid present only in Y. pseudotuberculosis strains causing clinical-epidemic manifestation of the infections as Far East scarlet-like fever (FESLF). The data obtained on the abihty of far-eastern strains to produce YPMa super-antigen, Y. pseudotuberculosis-derivative mitogen A, probably give evidence on its key role in FESLF pathogenesis. Variability of damage of innate immunity cells and target-organs caused by various plasmid types of Y. pseudotuberculosis by virulence could determine polymorphism of clinical-morphological manifestations of this infection. In-depth understanding of dependency of immune pathogenesis mechanisms of the disease on molecular characteristics of the causative agent opens up perspectives of enhancement of diagnostics and prognosis of the severity of the course of pseudotuberculosis and yersiniosis in human in general.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):74-85
pages 74-85 views

THEORETICAL ASPECTS OF CREATION OF SPECIFIC PREPARATIONS FOR ETIOTROPIC THERAPY AND PROPHYLAXIS OF WEST NILE FEVER AND CRIMEAN HAEMORRHAGIC FEVER

Efremenko V.I., Efremenko A.A., Efremenko D.V.

Abstract

Problematic issues on creation and practical introduction of specific immune biologic preparations for therapy and prophylaxis of natural-foci arbovirus infections - West Nile fever (WNF) and Crimean hemorrhagic fever (CHF), that are not available until now, are examined. Persistent natural foci of WNF and CHF with epidemic manifestations have formed in Southern and North Caucasian Federal Districts of Russia. Markers of Wfest Nile virus are being detected in central regions of Russia and Siberia, and the presence of fraction of population immune to this infection is also detected. Analysis of literature sources that has been carried out forms a theoretical basis for creation of novel specific preparations for etiotropic therapy and prophylaxis ofWNF and CHF. Use of blood from healthy donors with sufficiently high titers of class G immunoglobulins residing in certain subjects of Russian Federation in natural foci territories with the most intensive epidemic process is possible as raw material.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):85-93
pages 85-93 views

PLAGUE: CASES OF HUMAN INFECTION IN THE USA NATURAL FOCI AND THE DIFFICULTY OF THEIR DETECTION

Arutyunov Y.I.

Abstract

In this work we analyze the epidemic situations with plague in the United States of America, which are linked to internal migration - infection in a natural focus and movement of a diseased person within the boundaries of this focus to another district, state or non-endemic territory, as well as with international migration of infected person during incubation period.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):94-100
pages 94-100 views

DETERMINATION OF VIRULENCE PROPERTIES OF PATHOGENIC MICROORGANISMS IN VITRO: STATE-OF-ART

Shmelkova T.P., Sazanova E.V., Kravtsov A.L., Malyukova T.A., Popov Y.A., Boiko A.V., Devdariani Z.L., Schukovskaya T.N.

Abstract

Various methods for evaluation of virulence properties of causative agents of infectious diseases in vitro were analyzed: molecular-genetic, cultural-biochemical, immunologic, physiologic. Predominant use of molecular-genetic methods, expediency of a complex approach, relevance of search of novel informative parameters of virulence are noted. Study of biological properties of pathogens in vitro is the first screening stage of evaluation of their virulence.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):100-108
pages 100-108 views

INFECTIONS CAUSED BY EPSTEIN-BARR VIRUS IN HIV-INFECTED PATIENTS

Puzyreva L.V., Safonov A.D.

Abstract

The review is dedicated to features of clinical manifestations of infections caused by Epstein-Barr virus (EBV) in HIV-infected patients, problems of diagnostics and execution of antiviral therapy in the case of combination of these infections. Individuals at AIDS stage develop tumors, associated with EBV: non-Hodgkin’s lymphomas, including Berkitt’s lymphoma, primary B-cell lymphoma ofCNS, nasopharyngeal carcinoma. Formation of lymphoid interstitial pneumonitis and leukoplakia is known to be associated with EBV. A large list of preparations that are inhibitors of EBV replication are currently known, however, there is no clear pathogenetically justified therapy scheme for patients with this infection against the background of HIV-infection.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):108-116
pages 108-116 views

EBOLA FEVER

Markin V.A., Pantyukhov V.B.

Abstract

Problems of etiology, taxonomy and nomenclature of filoviruses, epidemiology, morbidity with a little-known by Russian medics especially dangerous exotic infectious disease - Ebola fever are examined. Significant distinguishing features of 2013 - 2015 epidemic in West Africa were detected - along with its unprecedented length, a decline did not take place as in previous outbreaks, neither causative agent virulence, nor infectivity of the infection during multiple generations from human to human. Literature data analysis allowed to assume that in the process of epidemic focus formation Ebola virus changes its properties and cyclically passes through several successive interconnected phases: an initial reservation phase in unknown ecosystems - animals, either plant, soil or water; intermediate phase of epidemic spread with primary acquisition of high virulence for humans, and then its decline; final stage of hidden circulation of causative agent that is ap-athogenic for humans. This hypothetical chain of natural phases’ transitions of Ebola virus allows to explain and link together phenomenology of this causative agent - rapid fall of virulence and infectivity for humans in foci in dynamics of epidemic outbreaks, quite a high population immunity in nosoareal of the causative agent in Africa, that contradicts the established understanding of its high lethality for humans.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):116-125
pages 116-125 views

OBITUARIES

НЕКРОЛОГИ

Article E.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):126
pages 126 views

СОДЕРЖАНИЕ

CONTENTS

Article E.
Journal of microbiology, epidemiology and immunobiology. 2016;93(6):127-128
pages 127-128 views


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