Vol 96, No 5 (2019)

ORIGINAL RESEARCHES

Development of the scheme of obtaining antibodies to the ribonucleoprotein of attenuated rabies virus

Gavrilova Y.K., Generalov S.V., Kireev M.N., Sharapova N.A., Abramova E.G., Savitskaya L.V., Ovchinnikova M.V., Kirillova T.Y., Semakova A.P.

Abstract

Aim. Isolation of ribonucleoprotein (RNP) of an attenuated rabies virus, develop schemes for immunizing animals with RNP-based preparations and determine the most effective scheme that allows obtaining serum with a high antibody titer to the RNP.

Materials and methods. We used the transplantable cell line Vero, the strain rabies virus «Moscow 3253 Vero», adapted for reproduction on Vero, rabbits of the chinchilla breed. In order to obtain serums containing antibodies to the RNP of the rabies virus, experimental schemes have been proposed for immunizing animals with RNP, including with adjuvants: polyoxidonium and colloidal gold. The dynamics of the accumulation of antibodies to the RNP of the rabies virus in the blood serum of experimental animals was studied by dot-immunoassay.

Results. The target component (RNP of the rabies virus) was isolated directly from the cytoplasm of the Vero cell culture infected with the rabies virus according to the modified M. Dastkhosh (2014) method, lyophilized and used in the development of preparations for immunizing experimental animals. In the study of the dynamics of the formation of antibodies to RNP of the rabies virus by the method of dot-immunoassay, the effectiveness of an adjuvant is established — colloidal gold nanoparticles ranging in size from 15 to 17 nm, the use of which makes it possible to increase the antibody titer by 2 times.

Conclusion. The results obtained are of interest for further research related to the design of diagnostic products and the development of methodological techniques using such preparations.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):3-8
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Development of genotyping method of the glanders causative agent based on multiple locus variable-number tandem repeat analysis

Bondareva O.S., Tkachenko G.A., Ledeneva M.L., Baturin A.A., Lemasova L.V., Shpak I.M., Budchenko A.A.

Abstract

The aim was to develop a short MLVA-typing scheme of the causative agent of glanders and to assess the possibility of its use for differentiation of Burkholderia mallei strains and study their genetic polymorphism.

Materials and methods. The study was carried out on 14 B. mallei strains from the collection of the Volgograd Research Institute for Plague Control and 12 whole genome sequences of the B. mallei strains presented in GenBank NCBI. A set of 32 loci proposed for differentiation of the melioidosis pathogen was used to select VNTR-loci for typing the causative agent of glanders. Polyacrylamide gel electrophoresis, sequencing, and fragment analysis were applied to detect the size of the VNTR fragments.

Results. VNTR loci 993, 3145, 3652, 20, 2862, and 1217, which were selected as the most variable among the causative agent of glanders, were included in the final MLVA typing scheme. The parameters of setting and detecting the results of MLVA typing have been optimized.

Conclusion. Analisys of the typing results of 26 B. mallei strains showed a high discriminating power of the developed method of intraspecies differentiation of glanders pathogen based on 6-loci MLVA-scheme and the prospects of its use for epidemiological investigation to determine the source of the glanders outbreak.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):8-16
pages 8-16 views

Inhibition of hemolytic activity of Streptococcus pyogenes in mechanisms of antibacterial action of zinc ions

Cheknev S.B., Vostrova E.I., Sarycheva M.A., Vostrov A.V.

Abstract

Aim. The work was performed with the purpose to study a hemolytic activity in the culture of S.pyogenes under the inhibitory action of millimolar concentrations of zinc ions.

Materials and methods. Suspensions of S.pyogenes bacteria which contained 108 CFU/ml were sown by the lawns into the standard Petri dishes coated with the supplemented Blood Nutrient Agar. 30 min later the salt solutions of zinc or copper which contained the metals at the concentrations ranged between 5 x 10-3 M to 5 x 10-1 M were added by the 5 μl drops on the surfaces of the lawns with use of 36-channel stamp replicator. Then the dishes with bacterial cultures were incubated for 24 hrs at 37°C followed by measuring diameter of the area of culture growth inhibition and of the area of inhibition of hemolysis. The study was performed with use of controls towards measuring the state of bacterial cells obtained from different zones of the areas.

Results. In presence of the zinc ions concentrations ranged between 50 to 500 mM the area of the growth inhibition of S.pyogenes was surrounded on the lawn of the bacterial culture by the area of the inhibition of hemolysis where the growth inhibition of S.pyogenes was not registered. Copper ions did not form such an area of the hemolysis inhibition.

Conclusion. Inhibitory action of zinc ions on the hemolytic S.pyogenes activity in the culture seems to be specific and reversible, and is discussed in a context of the antivirulent zinc ions properties.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):16-23
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The influence of seasonal influenza viruses biological features on the effectiveness of development strains for live influenza vaccine

Larionova N.V., Kiseleva I.V., Bazhenova E.A., Grigorieva E.P., Rudenko L.G.

Abstract

Aim. Evaluation of the efficiency of the method of reassortant strains for live influenza vaccine development and ways to optimize it, taking into account the differences in the current epidemic influenza viruses by key biological characteristics.

Materials and methods. Influenza viruses — candidates for seasonal LAIVs, MDVs for Russian LAIVs A/Leningrad/134/17/57 (H2N2) and B/USSR/60/69. The vaccine strains development in developing chicken embryos included reassortment, selective passages at low temperature in the presence of hyperimmune serum to the MDV, several stages of reassortants cloning, their virological and molecular genetic characteristics. Epidemic influenza viruses and LAIVs strains were evaluated by their ability to reproduction at temperatures beyond optimal values, by sensitivity to serum inhibitors.

Results. The assessment of phenotypic properties used in reassortment epidemic viruses is carried out. Presented the data on the efficiency of development reassortant strains for LAIV depending on the biological properties of circulating epidemic influenza viruses: their temperature-resistant, cold-sensitive phenotype, inhibitor resistance, and receptor specificity.

Conclusion. Based on the assessment of the influence of the biological characteristics of the epidemic viruses, the rational methodological techniques for the most effective development of reassortants for LAIV are selected.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):24-34
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The use of a polyvalent bacterial lysate in a complex therapy of chronic urticaria

Sorokina E.V., Sivakova N.G., Akhmatova E.A., Mitrofanova N.N., Shodova S.A., Akhmatova N.K.

Abstract

The aim of the study was to study trigger factors in chronic urticaria, peculiarities in the expression of Toll-like receptors, clinical and immunological efficacy of microbial antigens in patients with chronic urticaria.

Materials and methods. Patients with chronic urticaria (134 patients). A study of the expression of TLR2, TLR3, TLR4, TLR9 on blood cells using flow cytometry. 62 patients received polyvalent bacterial lysate against baseline therapy per os, 72 patients received monotherapy with basic drugs.

Results. In patients with bacterial infection, high levels of TLR2, TLR4 expression were detected. In the presence of viral infections, high TLR3 expression values were observed. The use of PBL contributed to an increase in the number of patients with clinical remission, decreased urticaria activity, led to a correction in TLR2 and TLR4, and decreased the level of total IgE.

Conclusion. Inclusion in the complex of therapeutic and prophylactic measures in patients with urticaria of a chronic drug based on microbial antigens (polyvalent bacterial lysate) contributes to the increase of clinical effectiveness and activation of the links of innate immunity.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):34-39
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Enterobacteria in the intestinal microbiocenosis of hivinfected children

Zakharova Y.V., Levanova L.A., Ivanov V.I., Bykov A.C., Afanasiev S.S., Afanasiev M.S., Karaulov A.V.

Abstract

Aim. The purpose was to assess the species composition, virulence factors and the nature of the interaction of Enterobacteria with intestinal microsymbionts in HIV-positive children.

Material and methods. The composition of intestinal microbiocenosis of 89 HIV-infected children (main group) and 74 children without HIV status (comparison group) was studied. 273 strains of Enterobacteria were isolated and identified to the species. The frequency and level of expression of adhesion, invasion, toxin formation and antagonism factors were studied.

Results. In HIV-positive children colonization rate of intestinal mucosa E. coli lac — was 5 times higher, E. coli hly+ 1.5 times higher than in the comparison group. The quantitative level of opportunistic enterobacteria increases with the aggravation of microecological disorders, 98% of enterobacteria forms a many-to-many association. Of these, 86.8% are neutral towards other symbionts. The frequency and level of production of adhesion, invasion and toxin formation factors did not differ in the compared groups.

Conclusion. In HIV-infected children is formed enterobacteriales type microbiota. Biological properties of enterobacteria did not depend on the presence of HIV status in children.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):40-46
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The biofilm formation of freshly isolated and vaccine strains of Bordetella pertussis of different serotypes

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

Abstract

Aim. To study the formation of biofilms by freshly isolated and vaccine strains of Bordetella pertussis of different serotypes.

Materials and methods. The intensity of biofilm formation by B. pertussis strains in 96-well round-bottom polystyrene plates by using three sowing doses of microbial cells (1,25 IOU/ml, 2,5 IOU/ml and 5,0 IOU/ml) was estimated by staining with 0,1% gentian-violet solution. The results were interpreted after measuring the optical density (OP) of the colored solvent at a wavelength of 600 nm.

Results. The highest intensity of biofilm formation was found in the newly isolated strain No. 211 and vaccine strain No. 475, both belonging to serotype 1.2.3. Cultures of these strains formed dense biofilms at all sowing doses of microbial cells. Certain differences in the intensity of biofilm formation were found between freshly isolated and vaccine strains of serotypes 1.2.0 and 1.0.3, especially when using a sowing dose of 5,0 IOU/ml. Freshly isolated strains at this dose formed dense biofilms, while two of the three vaccine strains formed moderate biofilms, and one strain was dense.

Conclusion. The revealed differences between the strains of B. pertussis in the intensity of biofilm formation may be associated with the peculiarities of the expression of agglutinogens, as well as other surface structures of microbial cells involved in the adhesion process on the substrate.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):47-50
pages 47-50 views

REVIEWS

Nosocomial respiratory viral infections: state of the problems

Akimkin V.G., Zakharova Y.A., Igonina E.P., Bolgarova E.V.

Abstract

We scanned the PubMed search database for literature on the incidence of nosocomial respiratory viral infections (NRVI) published over a ten-year period. Necessity to apply the standard case definition and the laboratory panel based on the multiplex polymerase chain reaction in frequency assessment was established. In general, predominance of rhinoviruses in the etiological structure was detected. Rationale was given for introduction of nonspecific epidemic prevention activities against a broad spectrum of other respiratory pathogens including high-priority respiratory syncytial viruses, metapneumoviruses, adenoviruses, influenza and parainfluenza viruses and coronaviruses. Bocaviruses and mimiviruses were designated as rare species. The biological diversity of the pathogens causing NRVI calls for active promotion of molecular genetics techniques in the work of the laboratory services of health facilities to perform quality etiological diagnosis, design relevant antiviral therapy regimens and effective prevention programs whose implementation will lead to significant reduction in the spread risk of these infections and the treatment costs.
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):50-61
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Mechanism of intracellular bacterial parasitism

Boichenko M.N., Kravtsova E.O., Zverev V.V.

Abstract

Algorithm of intracellular bacterial parasitism does not depend on if bacterium is obligate or facultative intracellular parasite. Depending on replicative niche’s localization intracellular bacterial parasites are divided onto cellular and vacuolated. Rickettsia spp., Shigella spp., Chlamydia spp. and Listeria monocytogenes use cell’s machinery of actin polymerization during process of their intracellular parasitism. These bacteria possess some of effector’s proteins which contain domains identical to effector proteins from the host cell. Shigella spp. T3SS and autotransporter protein IscA provide this process together with spreading bacteria intra colonic epithelium. In contrast other intracellular bacterial parasites, Listeria monocytogenes switches from dissemination in cytosol to persist in vacuole. In case of Brucella spp. the leading role in the creation of a replicative niche and in the modulation of the innate immune response is played by effector proteins of fourth type secretory system (T4SS).
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):61-72
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Efficacy of pneumococcal vaccine in immunocompetent and immunocompromised patients

Kostinov M.P., Kostinov A.M., Pakhomov D.V., Polishchuk V.B., Kostinova A.M., Shmitko A.D., Tarasova A.A.

Abstract

The article presents the analysis of numerous scientific studies carried out in Russia with the use of pneumococcal polysaccharide vaccine in immunocompetent and immunocompromised patients. New data are available to assess the impact of vaccines on clinical and immunological aspects in a particular pathology in children, which allows to reveal the mechanisms associated with the effectiveness of vaccination.
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):72-83
pages 72-83 views

Rabies and rabies immunobiological preparations: vaccinations Pasteur to the contemporary biotechnology

Abramova E.G., Nikiforov A.K., Movsesyants A.A., Zhulidov I.M.

Abstract

The review provides information on topical issues of rabies spread in the world and the Russian Federation, the stages of development and directions of improvement of available preventive anti-rabies immunobiological preparation used in medical practice for active and passive immunization against rabies. The current level of biotechnology development with the use of molecular biology and genetic engineering methods opens up prospects for the design of new safe effective anti-rabies drugs using recombinant technologies. Expanding the range of immunobiological drugs against rabies and their introduction into health practice will contribute to the elimination of human mortality from rabies.
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):83-94
pages 83-94 views

Genetic diversity of rubella virus at the present stage of rubella elimination and congenital rubella prevention programs

Chekhlyaeva T.S., Shulga S.V., Erokhov D.V., Tikhonova N.T., Zverev V.V.

Abstract

The review is devoted to an analysis of the genetic diversity of the rubella virus. Historical and geographical data of the circulation of various genotypes of the rubella virus were presented. The tendency to reduce the genetic diversity of the virus in the world as a result of the rubella elimination programs have been shown.
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):95-102
pages 95-102 views

Vaccine prophylaxis of intercurrent infections of respiratory tract in patients with tuberculosis

Mashilov K.V., Kostinova T.A.

Abstract

In this review, on the base of analysis of the current epidemiological data, the objective preconditions of the growing role of vaccinal prevention in increasing of effectiveness of treatment of tuberculosis in the key groups of patient is elucidated. Here are actual recommendations and information about efficiency and safety of vaccines in treatment of tuberculosis.
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):102-108
pages 102-108 views

Changing the spectrum of fatty acids as one of the mechanisms of adaptation/persistence of microorganisms

Shipko E.S., Duvanova O.V.

Abstract

The review presents the basic information available in the literature on changes in the composition of fatty acids in various microorganisms in response to various environmental factors (stressors). The issues affecting the importance of fatty acids as biomarkers of pathogenetic and adaptive-persistent potential of bacteria are discussed. The prospects of studying the spectrum of fatty acids in the field of biochemistry, in particular, lipidomics of infectious diseases are noted.
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):109-118
pages 109-118 views

Endemicity and enzooticity aspects of leptospirosis

Breneva N.V., Balakhonov S.V.

Abstract

The review covers the main definitions and criteria for endemicity and enzooticity relating to natural focal zoonotic infections and, in particular, to leptospirosis. Since leptospirosis is widespread, and almost all territories are endemic for this disease, it is proposed to allocate a high, medium and low degree of endemicity on the basis of several criteria, primarily the population morbidity. The unevenness of endemicity is also important in the epidemiological analysis of the leptospirosis situation.
Journal of microbiology, epidemiology and immunobiology. 2019;96(5):118-125
pages 118-125 views

BOOK REVIEW

О.В.Бухарин, А.А.Стадников, Н.Б.Перунова. Роль окситоцина и микробиоты в регуляции взаимодействий про- и эукариот при инфекции

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Abstract

О.В.Бухарин, А.А.Стадников, Н.Б.Перунова. Роль окситоцина и микробиоты в регуляции взаимодействий про- и эукариот при инфекции. Екатеринбург, УРО РАН, 2019, 247 с.

Journal of microbiology, epidemiology and immunobiology. 2019;96(5):125-126
pages 125-126 views


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