Vol 97, No 1 (2020)

The aim of this study was to obtain hybridomas producing monoclonal antibodies (Mabs) to the G-protein of the respiratory syncytial virus (RSV), and to evaluate their immunological characteristics and virus-neutralizing activity.

Material and methods. Mouse Mabs were obtained using hybridoma technology. The properties of Mabs were studied by enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining (IF) of infected cells, as well as by biological neutralization test in vitro (NT). To identify epitopes recognized by the Mabs on G protein ELISA additivity test was used.

Results. Hybridization of splenocytes with Sp2/0 myeloma cells and primary screening showed that 75 hybridomas produce antibodies interacting with purified virus, 17 of them also react with the recombinant G-protein in ELISA. In NT 4, hybridomas suppressed in vitro RSV infection by more than 50%. Cloning of these hybridomas revealed 4 monoclones producing the most active Mabs. Mab 1C11 was IgG2a, 3 others (5D4, 5G11 and 6H4) were IgM. Three IgM Mabs actively reacted with both RSV A2 and Long, and with G-protein; Mab 1C11 was less reactive with all antigens tested. All Mabs suppressed RSV infection, while Mab 5D4 supressed it almost completely (98%). IF analysis showed that all Mabs detected RSV G-protein in the cell cytoplasm, the largest number of infected cells was detected using Mab 5D4 (80%). It was shown that the isolated Mabs were directed to two non-overlapping epitopes on the RSV G-protein.

Conclusion. The isolated Mabs can be used to detect RSV in clinical samples by ELISA and IF. The isolated Mabs can be used for humanized recombinant antibodies construction and for the treatment of RSV infection in future.

Introduction. At present, the question of commensal, including opportunistic, microflora participation in  infertility development remains debatable. In a number of studies, the translocation of the vaginal microflora into the endometrial tissue is considered as a factor contributing to inflammation development. In addition, the connection of some reproductive losses with the persistence of certain conditionally pathogenic microorganisms is shown. Today, to solve this issue, molecular genetic research methods are being actively introduced that surpass the routine cultivation techniques in a number of positions.

The aim of the study was to assess the taxonomic diversity of microorganisms in the vaginal biotope with infertility.

Material and methods. For the study samples of vaginal contents obtained from the posterior vaginal fornix of 15 women, consisting of barren marriage, were used. A metagenomic study of 16S ribosomal RNA samples was carried out on the Illumina MiSeq platform, using the MiSeq Reagent Kits v3 kit (600-Cycle Kit), as recommended by the manufacturer. Libraries for sequencing plots of the V3-V4 gene of the 16S ribosomal RNA were prepared according to the 16S Metagenomic Sequencing Library Preparation Illumina. In bioinformatics assessment, Kraken Metagenomics version 2.0.0 software for metagenomic analysis (classifier of reads — short nucleotide sequences) was used using a standard database.

Results. It was shown that the occurrence of representatives of the Lactobacillaceae family in the vaginal biotope varies from 12 to 84%. The genus Lactobacillus with the dominance of L. jensenii, L. delbrueckii and L. amylolyticus occupied the leading position among the members of the family. In all samples, Moraxella spp. was found in large numbers, with M. osloensis leading among the representatives of the entire community. In half of the cases, the joint presence of M. osloensis and G. vaginalis was revealed.

Conclusion. It has been established that in infertility in the vaginal microbiota the number of representatives of the genus Lactobacillus is significantly reduced, and there is also a change in the leading species to L. jensenii, whose functional activity does not fully ensure the colonization resistance of the vaginal biotope, allowing for excessive reproduction of oppotrunistic microorganisms, in particular, M. osloensis.

Purpose. Study of the etiological structure of lethality from acute viral hepatitis and its comparison with morbidity and mortality for the period of 2009−2018.

Materials and methods. State Reporting Forms No. 1 “Report on Infectious, Parasitic and Non-Infectious Diseases” for the period of 2009−2018 were studied. The data was processed by the Microsoft Office Excel statistical package.

Results. In Kyrgyzstan, during the period of 2009−2018, 138,612 cases of acute viral hepatitis (AVH) were detected, of which 109 patients had fatal outcomes. In the general structure of the latter, the proportions of patients with acute hepatitis B (AHB 36.7%) and A (AHA, 35.7%) were approximately the same. In every fifth case, “unverified acute viral hepatitis (UAVH)” was diagnosed (22.9%). Acute hepatitis C (AHC) was the cause of death in 4 patients (3%), and hepatitis D virus (HDV) infection in 1 patient. Among the deceased, there were no cases of hepatitis E. Lethality from AHB was recorded in 5 children, and from hepatitis C in one child. At the same time, 9 out of 25 patients with “Unverified Acute Viral Hepatitis” were children. Over the period of 2009−2018, the incidence of AHB was 31.3 times lower than that of all AVH in general (240.9 and 7.7⁰/₀₀₀₀, AVH and AHB, respectively), the mortality rate was 2.7 times lower (0.19 and 0.07⁰/₀₀₀₀, AVH and AHB, respectively), and the lethality rate was 11.4 times higher (7.9 and 89.8⁰/₀₀₀₀, AVH and AHB, respectively). The average incidence of AHB in children was 6.4 times lower (7.7 and 1.2⁰/₀₀₀₀, total and children, respectively), the mortality rate was 2.3 times lower (0.07 and 0.03⁰/₀₀₀₀, total and children, respectively), and the lethality rate was 1.8 times higher (89.8 and 165.9⁰/₀₀₀₀, total and children, respectively).

Conclusion. The existence of morbidity and lethality in adults and children from acute hepatitis indicates a high disease burden for the country. The high proportion of AHB and AHA in the structure of AVH mortality requires improving the quality of immunization of children and expanding the coverage of adults with vaccination against these viral hepatitis. The increase in lethality against the background of a tenfold decrease in the incidence of AVH points out that the number of patients who have died from this pathology does not decrease.

Relevance. In September, 2017 the International Convention on Control of Ships’ Ballast Waters and sediments, in which the Russian Federation takes part, came into effect.

Aim of article is to cover the results of implementation of the Convention in Russia, regarding selection and analysis of ballast waters tests for compliance with the international standard.

Material and methods. The materials for work were data on ship arrivals at the international seaports of the Russian Federation, provided by sanitary and quarantine departments of the Russian ports, and monitoring researches of ballast waters in seaports of some regions of the Russian Federation. Analytical, bacteriological, molecular methods were applied.

Results. The studies of ballast waters in the Leningrad and Kaliningrad regions have been conducted for the first time in 2018, and in the Rostov region the study lasts since 2010. The laboratory researches of ships’ ballast showed that E. coli, Enterococcus spp. were in norm, V. cholerae O1 and O139 in ballast were absent. 12 of 21 ballast water tests investigated by specialists of the Rostov region laboratories contained V. cholerae non-O1/non-O139, ballast was taken on the ships which arrived from Romania and Turkey.

Сonclusion. The results of the molecular and genetic researches suggest that there is a probability of V. сholeraе introduction brougth with ship ballast. Management decisions are demanded to ensure biological safety of shipping and to decrease intestinal infections incidence in residents of the seaside cities.

The paper discusses the issues of morphofunctional variability of sapronoses pathogens in stressful environment. In the current century, sapronoses infections attract increasing attention. Under unfavorable environmental conditions the pathogens use the strategy for the formation of resting (stable) states, that is: viable but non-culturable cell formes and persistent bacteria, which are characterized by reduced metabolism and changes in the morphology and physiology of the microorganisms, termination of replication. Possibility of sapronoses pathogens survival in interepidemic period and antibiotic resistance formation, which play an important role in chronic infections, are associated with the formation of persistent forms of bacteria. The literature extensively discusses mechanisms and conditions of the pathogenic bacteria stable states formation and their pathogenetic contribution to infectious pathology, whereas ultrastructural organization and morphological variability of persistent cell forms, as well as their differentiation, causing the pathogens population heterogeneity, is still insufficiently illuminated. Based on the analysis of current data and their own experience, the authors evaluate the morphological and functional changes of bacteria stable cellular forms and their role in sapronoses pathogens adaptation strategies.

Features of the cellular immune response in the presence of a microbial biofilm are well described in the literature. Based on numerous studies, it became possible to establish a number of patterns: mature biofilms are better protected from immune factors, the effectiveness of antibiofilm strategies depends on species of the microorganisms, forming the biofilm, and, accordingly, on the composition of the biopolymer matrix. For example, rhamnolipids and alginate of Pseudomonas aeruginosa exert a significant negative effect on the function of immunocompetent cells. The bacteria of biofilms became able to turn to their advantage many of the protective reactions developed by the immune system and fixed evolutionarily, applying them for the growth and development of the microbial consortium.

The article represents the history of studies of human diseases caused by faсultative parasites of humans, animals and plants, attributed by microbiologist V.I. Terskych in 1958 to the third class of infections named sapronoses. The concept of sapronous pathogens ecology is one of priorities of Russian medicine. Since that time, facts and generalizations have been accumulated in the world and national science, allowing to revise the initial concept of sapronous infections of biota — from prokaryotes, plants and invertebrates to warm-blooded animals and humans. To decide the controversial issues of the sapronoses theory, epidemiological terminology and systematics of infectious pathology, it is necessary to integrate the knowledge and systemic participation of specialists in medicine and biology in studying this problem. The results of this cooperation will make it possible to clarify and form a unified theoretical concept of the population pathology of the biota based on the ecological approach and general biological regularities of life existence in the world.

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