Vol 96, No 4 (2019)


The prevalence of HIV infection in patients of general hospitals as a parameter for monitoring the system of epidemiological surveillance

Solonin S.A., Korabelnikova M.I., Bazhenov A.I., Kudryavtseva E.N., Kuzin S.N., Godkov M.A.


Aim. The aim of this study is to compare the prevalence of HIV infection in patients of large multidisciplinary hospitals in providing emergency and planned medical care and to evaluate the value of the long-term dynamics of this indicator as a quantitative characteristic of the epidemic process of HIV infection. Materials and methods. We retrospectively analyzed the results of examination for HIV infection of patients in the Sklifosovsky Research Institute for Emergency Medicine (RIEM) and Moscow Regional Research Clinical Institute (MONIKI) from 2008 to 2017. Long-term dynamics of the prevalence HIV infection were analyzed by using linear regression analysis. The intensity of trends (growth/decline) was assessed by the criteria proposed by V. D. Belyakov et al. (1981). Results. We examined for HIV infection 251 213 and 165 194 people in the research RIEM and MONIKI, respectively. For decade, the number of hospitalizations of HIV-infected patients to RIEM increased by 1.7 times, to MONIKI by 3.1 times. Prevalence of HIV infection in the patients of RIEM exceeded MONIKI ones from 3,2 to 6,0 times. At the same time, MONIKI showed a statistically significant trend towards an increase in hospitalizations of HIV-infected citizens (R2=0,8049, p=0,0004) with a pronounced average annual growth rate of 9,6% per year. Conclusion. We obtained results indicates a tendency to increase the number of recourses HIV-infected persons to general hospitals inMoscow andMoscow region. The most significant average annual increase in the prevalence over a decade period were recorded in the intensive care (+14.2%) and surgical (+9.8%) departments of MONIKI. Indicator groups of high risk of HIV infection in theMoscow region should be considered patients with urgent surgical pathology, acute exogenous poisoning of chemical etiology, as well as persons suffering from mental disorders.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):3-10
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Mass spectrometry analysis of protein blood extracts of animals with experimental brucellos

Ulshina D.V., Kovalev D.A., Ponomarenko D.G., Rusanova D.V., Berdnikova T.V., Evchenko A.Y., Bobrysheva O.V., Siritsa Y.V., Pisarenko S.V., Zhirov A.M., Kuznetsova I.V., Varfolomeeva N.G., Kulichenko A.N.


Aim. The aim of the present research was to study the possibility of direct detection of the causative agent of brucellosis in a biomaterial under experimental conditions via the MALDI-TOF MS method using Mass-Up program resources and a set of packages for open-source statistical software R. Materials and methods. We used laboratory mice infected with the causative agents of Brucellosis (strains B. melitensis 548, B. abortus 544, B. suis 1330) as models. Protein profiling was performed on a MALDI-TOF Microflex «Bruker Daltonics» mass spectrometer. Results. The bioinformatic-statistical approach used for analyzing MALDI-TOF mass spectra allows to carry out a direct detection of Brucella in the biomaterial; besides, it is possible to determinate their species via the identification of a group of biomarkers. Conclusion. It was experimentally confirmed that the protein profiles of the blood extracts of infected animals contain 11 markers, including 6 genus specific for Brucella spp., which can be associated with Brucella infection.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):11-18
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Proteomic profiling of Yersinia pestis strains circulating in the area of natural plague foci of North Caucasus and Transcaucasia

Koteneva E.A., Kotenev E.S., Kalinin A.V., Tsarеva N.S., Kot L.A., Zharinova N.V., Zaitsev A.A., Pechkovsky G.A.


Aim. To create a database of mass spectra of Yersinia pestis strains, which will differentiate the strains of the main and Caucasian subspecies of the plague agent by MALDI-TOF MS. Materials and methods. MALDITOF mass spectrometry was used to study 50 strains of Y. pestis, isolated on the territory of 7 natural plague foci of the Caucasus and Transcaucasia in the period 1950-2012. The removal of mass spectra of extracts of cells of Y. pestis was performed using the mass spectrometer Microflex LT «Bruker Daltonics». The results were processed and analyzed in FlexAnalysis programs, аnd MALDI Biotyper V. 3.0. Results. Тhis study showed that mass spectra have characteristic features that allow differentiating strains of the main (Y. pestis pestis) and subspecies (Y. pestis caucasica). Peaks characteristic of each subspecies were detected. The presence in Y. pestis caucasica subspecies peaks characteristic of the ancestral form — Y. pseudotuberculosis indicates the ancient origin of this group, which is consistent with the data of molecular genetic and WGS analysis given in other publications. Conclusion. Тhis work shows the possibility of applying the MALDI-TOF method of mass spectrometry for rapid differentiation of strains of the main subspecies Y. pestis pestis from the subspecies Y. pestis caucasica, which have different significance in the development and maintenance of the epizootic process in natural plague foci as well as different virulence for humans. Identification of the strain to the subspecies level requires carrying out culture and biochemical tests, which can take several days. The proposed method makes it possible to differentiate and obtain a result within half an hour after receiving a pure culture.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):18-25
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Immunophenotyping of blood cells of experimental animals immunized with Brucella abortus thermoextracts

Balakhonov S.V., Dubrovina V.I., Voitkova V.V., Korytov K.M., Barannikova N.L., Nikolaev V.B., Shkaruba T.T.


Aim. To study the subpopulational structure of blood cells of the experimental animals immunized with thermoextracts (TE) of Brucella abortus in L- and S-form. Materials and methods. Total 100 certified («Vector», Novosibirsk) outbred mice were immunized with B. abortus I-206 TE in L- and S-form in 20 μg protein dose. After 1, 3, 7, 14 and 21 days of observation the phenotypes (CD45, CD3, CD4, CD8, CD19, CD69) of blood cells were detected. Results. General regularities were revealed after injection of the experimental preparations. So, B. abortus TE in L- and S-form caused the immune response that increased granulocyte number and expression of early activation marker CD69 by T- and B-lymphocytes of blood in early period of observation (1-3 days), decrease in general B-lymphocyte content in late periods of observation (7-21 days). Thus, mice received B. abortus ТE in L-form demonstrated authentically higher CD69 expression of blood lymphocyte subpopulations than mice received B. abortus ТE in S-form. Distinctions in formation of humoral immune response were revealed that probably was connected with alteration of Brucella chemical composition in the course of L-transformation. Conclusion. The investigation established that B. abortus TE in L- or S-form caused immunological reorganization in the experimental animal organisms. On the basis of the fin

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):25-31
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Serological methods for detection of the causative agent of tularemia and their evaluation

Zharnikova I.V., Efremenko V.I., Zharnikova T.V., Kurcheva S.A., Kalnoy S.M., Еfremenko D.V., Isakova A.A., Indenbom A.V.


Aim. A comparative study of serological methods for the detection of the causative agent of tularemia and their evaluation. Materials and methods. We used experimental diagnostic kits and test systems for the production of serological methods: indirect hemagglutination reaction (RGA); the reaction immunofluorescence (RIF); enzyme immunoassay (ELISA) using traditional microplate; IFA after selective concentration of the pathogen of tularemia in magnoimmunosorbents (MIS); microgravimetric analysis (MGA) based on piezoresistors (SP) and surface plasmon resonance (SPR). The experiments were carried out with homologous strains of tularemia microbe (test strains) and with strains of heterologous microorganisms in model experiments on tap water contaminated with different concentrations of the pathogen. Results. The parameters of each diagnostic method are determined and evaluated according to the following indicators: sensitivity (when working with pure cultures (test strains), contaminated samples of large volumes), specificity, time of setting and taking into account the results, informativeness, determining the modes of setting and accounting. Conclusion. The above diagnostic methods have their advantages and disadvantages. Therefore, when choosing a method, the researcher should be guided by the goals pursued. So, for screening studies it is advisable to carry out the formulation of ELISA, RIF, RGA, in identifying the pathogen in large volumes and contaminated samples, the effective use of selective concentration on MIS followed by the formulation of ELISA, to identify small amounts of samples and take into account the reaction in real time, it is possible to use MGA and SPR.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):32-38
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Molecular-genetic study of the RA-27/3 strain used for production of rubella vaccine

Ignatev G.M., Atrashevskaya E.V., Suchanova L.L., Sidorenko E.S., Netesova N.A.


Aim. In order to study rubella virus strain RA-27/3 genetic stability, used for the vaccines production, a molecular genetic study was conducted. Materials and methods. In the study different series of master and work seed of RA-27/3 rubella virus strain by «Microgen», a few lots of rubella vaccines by the different manufacturers, as well as strain «Orlov» of rubella virus were used. RT-PCR followed by restriction, sequencing were performed . Results. Full-genomic sequences of the rubella virus strain RA-27/3 by «Microgen», were obtained and presented to GenBank. The full structure correspondence of RA-27/3 rubella virus strain by «Microgen» to the similar rubella strains used by GSK and Merck & Co Inc. has been shown. The RT-PCR method with the subsequent restriction was fulfilled using only domestic reagents. The developed method has been demonstrated as applicable for the identification of the RA-27/3 rubella virus strain as in monopreparation as well as in the combined vaccine preparation. Conclusion. The data obtained make it possible to suggest application of the molecular genetic methods for the vaccine virus identification not only at the production stages, but also in the finished vaccine lots.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):38-46
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The modern condition of Crimean-Congo hаemorrhagic fever natural focus in the Russian Federation

Vasilenko N.F., Manin E.A., Maletskaya O.V., Volynkina A.S., Prislegina D.A., Semenko O.V., Kulichenko A.N.


Aim. To determine the boundaries of the Crimean-Congo haemorrhagic fever (CCHF) natural focus in the Russian Federation at the current stage, to clarify the range of the main reservoirs and vectors of CCHF pathogen, to assess the epidemiological capacity of the natural focus. Materials and methods. The materials of epidemiological and epizootological monitoring of the CCHF natural focus, methods of epidemiological and epizootological analysis, molecular-genetic and cartographic methods were used in the work. The findings have been treated using by software package Microsoft Office Excel 2010. Results. The unified integrity of the CCHF semi-desert-steppe natural focus, which occupies vast territory of the southern part of the Russian Federation of 831 thousand square kilometres, is science-based. Expanding the geographic area of the CCHF agent with the involvement new administrative district can be seen. The trend of shifting of the CCHF natural focus borders in a northerly direction has been established. An increasing of epidemiological capacity of the CCHF natural focus has been noted. Hyalomma marginatum ticks are the main reservoirs and vectors of CCHF virus. The genotype «Europe-1» is predominant genotype in the natural focus. Conclusion. It is necessary to improve the tactics of CCHF epidemiological surveillance using modern science-based approaches. For example, automated forecasting-modeling system, using results of multifactorial risk analysis, which have an impact on the intensity of CCHF epidemic appearances, allows to quantitative forecast epidemiological situation on this infection in the aggregate and for certain subjects of the south of the Russian Federation.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):46-52
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Prevalence of carbapenemase-producing Klebsiella pneumoniae in the Gomel Region

Tapalski D.V., Petrovskaya T.A., Bonda N.A., Kozlova A.I., Osipkina O.V.


Aim. Within the microbiological monitoring program, to study the prevalence of carbapenemase-producing K. pneumoniae in the healthcare organizations of the Gomel region and assess their level of resistance to antibacterial drugs. Materials and methods. For 91 clinical isolates of Klebsiella pneumoniae with multiple antibiotic resistance, isolated in Gomel and Gomel region, carbapenemase genes were detected by real-time PCR and sensitivity to antibacterial drugs was determined. Results. 68 carbapenemase producers were revealed: KPC — 1 isolate, OXA-48 — 47 isolates, NDM — 20 isolates. Carbapenemase producers were found in 11 Gomel health organizations and 8 central district hospitals of the regional centers of the Gomel region. All of them had an associated resistance to most antibiotics and retained sensitivity to colistin (91.2% sensitive isolates) and tigecycline (98.5%). Conclusion. The spread of carbapenemase-producing K. pneumoniae isolates in healthcare organizations makes it very difficult to conduct effective antibiotic therapy for patients and requires the introduction of appropriate infection control measures aimed at limiting their circulation in the hospital environment.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):53-58
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Severe cases of seasonal influenza in Russia in 2017-2018

Svyatchenko S.V., Durymanov A.G., Kolosova N.P., Gudymo A.S., Goncharova N.I., Torzhkova P.Y., Bulanovich Y.A., Epanchintseva A.V., Danilenko A.V., Marchenko V.Y., Sysoeva A.V., Susloparov I.M., Tregubchak T.V., Ryzhikov A.B., Maksyutov R.A., Ilicheva T.N.


Aim. Evaluation of herd immunity prior to the 2017-2018 influenza season, and characterization of influenza viruses isolated from severe or fatal influenza cases and from influenza cases in people vaccinated in the fall of 2017. Materials and methods. Evaluation of herd immunity in hemagglutination inhibition assay. Isolation of influenza viruses. Antigenic and genetic analysis. Results. Prior to epidemic season 33-47% of blood sera samples collected on the territory of Russia showed presence of protective antibody titers against vaccine strains of influenza A, 24-30% of samples — against B/Victoria. During 2017-2018 epidemic season 87 influenza A and B viruses were isolated. A(H1N1)pdm09 strains belonged to clade 6B.1, B/Yamagata strains to clade 3, and B/Victoria strains to clade 1A; they were antigenically similar to corresponding vaccine strains. A(H3N2) viruses belonged to clade 3C.2a and were difficult to characterize antigenically. One strain of influenza virus А(H1N1pdm09) was resistant to oseltamivir and had H275Y amino acid substitution in neuraminidase. All other isolates were susceptible to neuraminidase inhibitors. Conclusion. Influenza vaccination with vaccine effective against current circulating strains and treatment with neuraminidase inhibitor drugs at first manifestation of clinical signs of influenza disease are effective means of population protection against influenza.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):58-64
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Study of the effect of polyelectrolytes with antiviral effect on the activity of influenza virus neuraminidase and the process of oxidative phosphorylation in mitochondrias of cells host organism

Kontarov N.A., Pogarskaya I.V., Yuminova N.V.


Aim. Study of the inhibitory effect of polyelectrolytes with antiviral effect against influenza neuraminidase. Determination of the type and inhibition constant. The study of the effect of polyelectrolytes (PE) on the process of oxidative phosphorylation in the mitochondria of cells host organism. Materials and methods. Purified influenza virus strains were used: A/VPCH/Weybridge (H7N7), A/Mallard Pennsylvania/10218/84 (H5N2), A/NIBRG-14 (H5N1) with an initial infectious titer of 4,5 lgTCD50/ml. PE solutions of polystyrenesulfonate with a degree of polymerization of 8 (PSS-8) in concentrations of 0,5—4,0 mM and polyallylamine (6 kDa) PAA (6 kDa) in concentrations of 0,5—4,0 μM. To determine the activity of influenza neuraminidase, influenza virus strains were used after the removal of low molecular weight inhibitors of neuraminidase by dialysis against water. The neuraminidase substrate was fetuin at final concentrations of 0,052 to 1,2 μM for PAA (6 kDa) and from 0,052 to 1,2 mM for PSS-8. As quantitative characteristics of respiration and phosphorylation of mitochondria, respiratory coefficients according to Lardi-Velman and Chans-Williams, as well as the ratio of ADP/O were used. Mitochondria were isolated from skeletal muscle. The determination of respiratory coefficients and the ratio of ADP/O) was determined by the polarographic method. Results. A noncompetitive type of inhibition of these PEs was detected in relation to the neuraminidase activity of influenza viruses with inhibition constants KI = 1,6 ± 0.08 μM for PAA (6 kDa) and KI = 1,7 ± 0.085 mM for PSS-8. Respiratory coefficients and the ratio of ADP / 0 were determined in the absence and after addition of PSS-8 and PAA (6 kDa) to mitochondria at concentrations of 20 mM and 10 μM, respectively. A decrease in respiratory coefficients and an ADP/O ratio was observed, indicating an inhibition of the enzymes of the electron-transport chain of mitochondria. At concentrations of less than 20 mM and 10 μM for PSS-8 and PAA (6 kDa), all indicators did not significantly change. Conclusion. The non-competitive mechanism of inhibition of the neuraminidase activity of the influenza PE viruses is explained by the conformational changes in the molecules of the enzyme and/or enzyme-substrate complex and, accordingly, the structural and functional changes in its secondary structure. When going beyond the range of non-toxic concentrations of20 mM for PSS-8 and 10 μM for PAA (6 kDa), inhibition of the mitochondrial respiratory chain enzymes was observed while maintaining the antiviral effect.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):64-68
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Models of dependence of the quantity of the Streptococcus pneumoniae biomass and his capsular polysaccharide from the composition of the feeding environment

Smirnova A.O., Baranovskaya S.A., Tokarskaya M.M., Elkina S.I., Yastrebova N.E.


Aim.The development of a semi-synthetic nutrient medium that provides the maximum amount of capsular polysaccharide (CPS). Materials and methods. We used the strain 521 of S. pneumoniae serotype 23F. Cultivation was carried out in test tubes with 10 ml of polysynthetic nutrient medium of a specific composition. The amount of polysaccharide in the samples was determined using rocket immunoelectrophoresis. Building models and comparing the effects of various components was carried out according to the methodology specified in the tutorial. The calculation of the coefficients of the equation and the assessment of the adequacy of the equations themselves was carried out using RStudio version 1.0.153. Results. As a result of a series of experiments, the coefficients of the regression equations were calculated, their significance was evaluated, and models of dependence of biomass production and CPS were constructed depending on the composition of the nutrient medium. To solve the problem, an experiment was carried out according to the Box-Wilson method. The peptone and glucose concentrations were selected as optimized parameters. The step size ΔSi in the increasing gradient direction was calculated based on the coefficients of the regression equation. At the same time, the exact nature of the dependence was determined. The optimal calculated concentrations of peptone and glucose, at which the formation of CPS is maximum, are 32.6 and 12.1 g/l, respectively. In this case, the forecast yield of the polysaccharide is 239 mg/l. Conclusion. Using the method of fractional factorial experiment, models of the dependence of the biomass amount of S. pneumoniae and its capsular polysaccharide on the composition of the nutrient medium were obtained. The optimal concentrations of the components of the medium were found, which make it possible to increase the level of biomass formation by 10% compared to the standard formulas, and the CPS — by 1.5—2 times.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):68-72
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Comparative analysis of efficiency of bacteriological and molecular-biological methods for the assessment of microbial contamination of hospital environment objects

Orlova O.A., Semenenko T.A., Akimkin V.G.


Aim. To conduct a comparative analysis of bacteriological and molecular biological research methods to assess the microbial contamination of objects of hospital environment as factors of microbial transmission. Materials and methods. A comprehensive laboratory investigation of nosocomial environment of surgical, intensive care, hematology divisions of a large multi- disciplinary clinic during 2017 was carried out, while 215 samples were selected. Results. Microbial contamination of hospital environment facilities accounted for 54,0% when carrying out bacteriological tests and 80,0% — in conducting molecular biological studies. Between the frequency of bacteriological isolation of microorganisms and the determination of DNA of pathogens in different departments of the hospital, a strong direct correlation was revealed (r=0.92). The coincidence of the results of the two methods of research in most cases was observed at high values of the number of DNA copies (800-10000). The molecular biological method of diagnosis revealed the Methicillin-resistant Staphylococcus 1.9 times more often and in 19,4±7,2% lactamase (MBL) genes in major gram-negative pathogens. Conclusion. The use of molecular biological methods allows to detect microorganisms on the objects of hospital environment much more often than during bacteriological examination.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):73-78
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Varicella Zoster virus infection: immunity, diagnosis and modelling in vivo

Lavrov V.F., Svitich O.A., Kazanova A.S., Kinkulkina A.R., Zverev V.V.


Varicella Zoster Virus (VZV) is a highly contagious viral agent of the Herpesviridae family, which has a strict species specificity, and causes two different diseases — chickenpox, mainly in children, and herpes zoster — more often in the elderly. Obtaining additional information about the life cycle of the virus, its biology, pathogenetic features of the complications caused by it, will contribute to the emergence of more advanced methods of diagnosis and prevention, the development of new experimental approaches that allow to study the innate and adaptive mechanisms of immune protection in VZV-infection.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):82-89
pages 82-89 views

Vaccination of patients with bronchial asthma against influenza and pneumococcal infection

Protasov A.V., Andreeva N.P., Kostinovа A.M.


The review presents data on the role of respiratory agents in the pathogenesis of exacerbations of bronchial asthma and the impact of vaccination on the clinical course of the disease. The features of the formation of protective immunity in various immunization schemes are analyzed. In patients with bronchial asthma, the substantiation of the vaccination scheme with the advantage of priority administration of conjugated polysaccharide vaccine followed by the introduction of polysaccharide pneumococcal vaccine after 8 weeks is given. It was shown that vaccination against pneumococcal infection resulted in elimination from sputum S. pneumoniae. Influenza vaccination in patients with bronchial asthma reduced the frequency and duration of exacerbations. Combined vaccination against influenza and pneumococcal infections did not reduce the clinical and immunological effect.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):90-98
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Genetic variability of Bordetella pertussis and its role in vaccine prevention of pertussis

Bazhanova I.G., Britsina M.V., Mertsalova N.U., Ozeretskovskaya M.N.


In many countries of the world despite the extensively vaccination against pertussis has increased the incidence of the whooping cough in all age group of the population. The MLST, MLVA and CGH investigations revealed the differences in genotypes between the vaccine strains B.pertussis and the circulating isolates B.pertussis in consequence of adaptation of the bacterium B.pertussis to the immunized hosts. It is lead to waning immynity and outbreak of incidence of pertussis. The mutations in the genes encoding the major virulence factors, the allelic polimorfism and decreasing the genome size of B.pertussis strains are the basis of the B.pertussis adaptation to the immunized hosts and dependent on the type of the vaccine used for immunization Programme. In countries that use acellular pertussis vaccine for vaccination programme the dominant isolates genotypes are: ptxА1-ptxС2- ptxР3-prn2- tcfA2-1-fim3-2 и ptxА1- ptxС2- ptxР3-prnА2- tcfA2- fim2-1- fim3-1, and that use the cellular pertussis vaccine the dominant isolates genotypes are ptxА1-ptxС1- ptxР1-prn1- tcfA2- fim2-2 fim3-1 и ptxА1- ptxС1- ptxР1- prn2- tcfA2- fim2-1- fim3-1. The constant monitoring of the genotypes of isolates B.pertussis is necessary to reveal the dominant genotypes and include them in the national immunization programme in combination with vaccine strains B.pertussis.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):98-105
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Akkermansia muciniphila is a new universal probiotic on the basis of live human commensal gut bacteria: the reality or legend?

Shenderov B.A., Yudin S.M., Zagaynova A.V., Shevyreva M.P.


Contemporary information on biology of Akkermansia muciniphila and the role of these gut mucolytic anaerobic bacteria in physiological functions, metabolic and signaling reactions in human health and diseases are presented in the review. Established by foreign researchers, favorable and negative effects are associated with the presence in these gram-negative bacteria specific surface membrane proteins, the production of certain short-chain fatty acids and endotoxin, as well as with the ability degrading mucin, changing intestinal barrier function and synthesizing some neurotransmitters. Prospects and difficulties of creation of new microbial nutraceuticals and drugs on the basis of living cells of A. muciniphila or their specific low-molecular components and metabolites are considered.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):105-115
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Molecular-genetic mechanisms of conservation of the pathogenic potential of the causative agents environments of natural-focus sapronosis

Andryukov B.G., Somova L.M., Bynina M.P., Lyapun I.N.


For interepidemic periods of natural focal sapronoses, various ways of maintaining the viability of pathogens in terrestrial parasitic systems are associated with various adaptation strategies necessary for the conservation of the population. Unlike spore-forming bacteria, sapronose pathogens use stable cellular forms — a viable but uncultivated state and persistence. The implementation of these strategies is due to the influence of various stress factors of the habitat and is characterized by a decrease in metabolism, a change in the morphology and physiology of the bacterial cell, and the cessation of its replication. It is important that stable forms of cells retain virulence and, when favorable conditions come, they are again transformed into active vegetative forms. The discovery in recent years of genetic modules of bacterial toxin-antitoxin systems has made it possible to uncover complex regulatory molecular mechanisms for preserving the pathogenic potential of stable forms of pathogens of natural focal sapronoses in interepidemic periods.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):115-126
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Manufacturing of hybridomas, producing monoclonal antibodies against Burkholderia mallei and Burkholderia pseudomallei antigens

Kuklina G.V., Elagin G.D., Pechenkin D.V., Fomenkov O.O., Eremkin A.V., Kytmanov A.A., Shurupov S.A., Ipatov S.S.


Aim. Obtaining hybridomas, stable producing specific monoclonal antibodies against Burkholderia mallei and Burkholderia pseudomallei antigens. Materials and methods. The microbial cultures from State Collection of Microorganisms from the Branch of 48 CSRI of the Defense Ministry of Russian Federation (Kirov) and BALB/c mouse were used in research. Hybridization of B lymphocytes with SP2/0-Ag14 myeloma cells was performed by G.Kohler and C.Milstein procedure in De St. Fazekas and D.Scheidegger modification. The specific activity of immune sera, hybridoma supernatants, ascites and evaluating the diagnostic capabilities of monoclonal antibodies was studied by ELISA. Results. Hybridomas, producing monoclonal antibodies against causative agents of glanders and melioidosis antigens, were obtained and characterized. Obtained hybridomas are active and stable antibody producers after repeated in vitro and in vivo passaging. Immunoglobulins from obtained ascites were isolated. Antibodies provided the greatest sensitivity and specificity were selected. Conclusion. Monoclonal antibodies, producing by obtained hybridomas may be used for creating of immune biological tests.

Journal of microbiology, epidemiology and immunobiology. 2019;96(4):78-82
pages 78-82 views

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