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Problems of Particularly Dangerous Infections

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No 1 (2022)
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REVIEWS

6-14 663
Abstract

The review briefly outlines the current taxonomy of the most studied strains of the species belonging to the family Francisellaceae. Due to the significant genetic diversity within the family, the detection of Francisella tularensis, especially in environmental samples, becomes even more difficult and may lead to false positive results. A comparison of the incidence rates in European countries, in which the extensive epidemic manifestations of tularemia are recorded annually, namely Sweden, Finland, Norway and the Czech Republic, and in Russia over the past five years is provided. The paper presents the comparative dynamics of tularemia incidence in the territories of the Federal Districts of the Russian Federation in recent years. The dynamics of the incidence during epidemic outbreaks in the Republic of Karelia and the Omsk Region over the past few years is shown. 17 cases of human infection with the causative agent of tularemia were registered on the territory of the Russian Federation in 2021. Epizootic manifestations of the infection of varying degrees of intensity were detected in 45 constituent entities of the Federation. Against that background, sporadic cases of tularemia in humans were reported in 11 regions of the country. Six cultures of Francisella tularensis subsp. mediasiatica from different types of ticks were isolated in the Krasnoyarsk Territory, the Republic of Altai and the Altai Territory. Based on the analysis of the data presented, in 2022, epidemic complications in the form of sporadic cases of the disease are most likely to occur among the unvaccinated population in the territories of the Central Federal District – the Voronezh, Ryazan, Smolensk Regions and in Moscow; Northwestern Federal District – in the Arkhangelsk Region, the Republic of Karelia and in St. Petersburg; Volga Federal District – in the territories of Samara, Kirov Regions and Republic of Tatarstan; Ural Federal District – in the Khanty-Mansiysk and Yamalo-Nenets Autonomous Districts; Siberian Federal District – in certain districts of the Kemerovo, Novosibirsk, Omsk and Tomsk Regions, Altai and Krasnoyarsk Territories.

15-23 632
Abstract

The aim of the work was to analyze the epidemiological situation on tick-borne viral encephalitis in the endemic territories of the Russian Federation in 2021 in comparison with the dynamics over 2011–2020 and its shortterm forecast for 2022. In Russia, 48 constituent entities belonging to seven federal districts are endemic for tick-borne viral encephalitis (TBVE). There is a statistically significant downward trend in the incidence of TBVE in the Siberian Federal District (which is characterized by the maximum incidence rate in the country), the Volga and Far Eastern Federal Districts. In the Ural Federal District (the second in terms of TBVE incidence), the decline in the incidence has stalled since 2021. The average long-term incidence of TBVE in the constituent entities of the Russian Federation varies from sporadic (Republic of Crimea)  up to 12.5 per 100 thousand of the population (0/0000) (Krasnoyarsk Territory). In 2021, TBVE cases were detected in 42 endemic regions and in one non-endemic region – Stavropol Territory. At the same time, 1015 cases of TBVE were in the country (0.69 0/0000). In all Federal Districts, the incidence of TBVE is below the long-term average values. Using the Quantum GIS program, the incidence of TBVE in 917 administrative territories of the country has been ranked and grouped according to the level of epidemiological risk. This made it possible to establish that 65 % of the territories form a zone of low epidemiological risk. High and very high epidemiological risk is observed in 13% of the analyzed districts. The structure of TBVE clinical manifestations in 2021 was dominated by febrile (59.7 %) and meningeal (24.3 %) forms. 14 lethal outcomes were reported. In 2021, 2 889 515 people were vaccinated (including 1 433 850 children), of which 14 fell ill. Specific immunoglobulin was used to prevent the overt development of infection in 100 704 individuals, which accounts for 22.6 % of the persons affected by tick bites (30.6 % among children). Acaricidic treatments were carried out on an operational area of 233 125 hectares of territories of socially significant objects. The scope of all TBVE prevention measures in 2021 increased as compared to 2020. In 2022, a decline in the incidence of TBVE in endemic Federal Districts and in the country on the whole is forecasted to (0,64±0,192) 0/0000.

24-34 1514
Abstract

The aim of the work was to summarize the results of cholera monitoring in 2021, to assess current trends in the development of the epidemic process, and to predict the epidemiological situation in the Russian Federation for 2022. It was established that within the period of 2012–2021, 4117264 cases of cholera with the spread of infection across 83 countries on all continents were registered in the world and there was a downward trend in the incidence in Asia and Africa. The dynamics of monthly morbidity in 2021 was associated with emergencies as factors of epidemiological risk. Epidemics and outbreaks of cholera were documented against the background of COVID-19 pandemic and laid a double burden on healthcare systems. At the same time, based on the overview of the results of cholera monitoring in the constituent entities of the Russian Federation, it was shown that the forecast of epidemic well-being given for 2021 was fully justified. It has been determined that the increase in the number of non-toxigenic strains of Vibrio cholerae O1 (67) isolated from water bodies compared to 2020 (25) is mainly due to the appurtenance of a number of isolates to clonal complexes. The study of phylogenetic relation has demonstrated that the detection of strains with genotypes which were previously identified in the isolates evidences the persistence potential. The identification of strains with new genotypes, which were earlier established in the strains circulating in other territories, pointed at the possibility of the occasional importations. The forecast of the epidemiological situation on cholera in Russia for 2022 is associated with the continuous existence of risks of introduction. If these epidemiological risks are not realized, a favorable epidemiological situation is predicted regarding this infection in the country. It is expected that the detection of epidemiologically insignificant strains of V. cholerae O1 in environmental water bodies, along with their clones and/or clonal complexes, will remain, including strains that may be an etiological factor in sporadic cases or outbreaks of disease.

35-42 646
Abstract

The aim of the research was to assess epizootic activity of natural plague foci in the Russian Federation in 2021 and provide a prognosis for 2022. Activation of epizootic process has been detected in Central-Caucasian high-mountain natural plague focus after a recess since 2007. Local epizootics persist to develop in Tuva Mountain and Gorno-Altai highmountain plague foci. Total area of epizooties in Russia in 2021 amounted to 1649.5 km2. 28 plague microbe strains were isolated. Animals infected with plague were not found in Tersko-Sunzhensky low-mountain, Dagestan plain-piedmont, Precaspian north-western steppe, Volga-Ural steppe, Trans-Baikal steppe, Volga-Ural sandy, Caspian sandy and EastCaucasian high-mountain natural plague foci. It is demonstrated that epidemiological welfare as regards plague in Russia in 2017–2021 was achieved through annual epizootic monitoring of plague-enzootic territories, timely epizooty detection and preventive activities aimed at reducing the risk of infection. Substantiated has been the forecast for tense epidemiological situation on the territory of Karachay-Cherkessia, Kabardino-Balkaria, Altai Republic and Tuva Republic in 2022. A new Epitracker application has been proven to be prospective for rapid assessment of actual epidemiological situation in natural plague foci.

43-53 891
Abstract

The epidemiological situation on West Nile fever (WNF) in Russia in 2021 was characterized by an increase in the incidence relative to 2020 (more than 6-fold increase). The peculiarities of the WNF epidemic process have been determined: the territorial distribution of cases (75 % – in the constituent entities of the Central Federal District), the expansion of the causative agent’s areal with the involvement of new territories (official registration of cases in Moscow and the Tula Region for the first time ever), the early end of the epidemic season. An increase in the proportion of neuroinvasive forms, an increase in the share of male patients and the age group of 30–39 years, a decrease in the proportion of the population in contact with the pathogen in natural places of mass recreation were observed in the incidence structure. According to the results of the monitoring studies carried out by the Reference Center, intensive circulation of the pathogen was established on the territory of the Central Federal District, Southern Federal District, and the North Caucasian Federal District. The low reported incidence was due to insufficient detection of WNF patients. The results of moleculargenetic study showed that in the European part of Russia, WNV lineage 2 circulated in both epizootic and epidemic cycles, lineage 2 in the south of Western Siberia (Omsk Region) and lineage 4 in the Volgograd Region – in the epizootic cycle only. Phylogenetic analysis revealed that strains isolated from the Astrakhan, Volgograd, Rostov, Voronezh Regions and the Republic of Dagestan in 2021 belong to the genovariant of WNV lineage 2 not registered in Russia previously.  The topology of the phylogenetic tree indicates the possible African origin of the isolates, probably imported into Russia by migratory birds across the coast of the Caspian Sea. In the Volgograd Region, WNV of both the entrenched clade of lineage 2 (since 2007) and new genovariant circulated. The specialists of the Reference Center developed a forecast of WNF epidemiological situation development in the Volgograd Region in 2022 on the basis of the neural network modeling technique, according to which an increase in the incidence is expected, comparable to that in 2010 and 2012. Based on the climate projections, an increase in the incidence is possible in all Federal Districts of the European part of Russia, the southern territories of the Urals, Western Siberia, and the Far East.

54-63 495
Abstract

The aim of the review was to characterize the epidemiological situation on rickettsiosis of the tick-borne spotted fever (TSF) group in the constituent entities of the Russian Federation in 2012–2021 and provide a long-term forecast for 2022–2026. In 2020–2021, indicators of registered incidence of tick-borne rickettsiosis in Russia on the whole decreased relative to the average long-term pre-pandemic level in 2012–2019 by 2 fold for all forms of the TSF group on average (Siberian tick-borne typhus – STT, Astrakhan spotted fever and Mediterranean fever). A comparative analysis of the territories revealed a correlation between the incidence of STT and the number of medical facility visits due to “tick bites” (r=0.67, p=0.015), which, against the background of an increase in the frequency of contacts of the population with vectors in 2020–2021 in most regions and a decrease in the incidence in the same regions indicates the credible nature of the changes. The ranking of territories according to average long-term incidence rates made it possible to classify the Republic of Altai and the Altai Territory as regions of extremely high epidemic hazard as regards STT; high hazard – the Republic of Tuva; above average – the Republic of Khakassia, Khabarovsk Territory and the Jewish Autonomous Region. The established downward trend in the incidence of tick-borne rickettsiosis among the population will be sustained in most entities of the Russian Federation within the next 5 years, excluding the Republic of Altai, the Altai Territory, the Republic of Crimea and the city of Sevastopol, where current incidence or an increase should be expected. The results of the analysis indicate the need to put more emphasis on natural-focal infections at the time of COVID-19 pandemic.

64-70 1433
Abstract

A generalized analysis of the epidemiological and epizootiological situation on anthrax in the world in 2021 is presented. Provided is the forecast for the Russian Federation for 2022. In 2021, two cases of cutaneous form of anthrax were recorded in Russia among population living in the Siberian (the Republic of Tuva) and the North Caucasian (the Republic of Dagestan) Federal Districts. Epizootiological and epidemiological instability due to the infection was manifested in neighboring countries – Azerbaijan, Armenia, Kazakhstan, Kyrgyzstan, and Ukraine. Anthrax among farm and wild animals was registered in a number of countries in Asia, Africa, Europe, North and South America, and Australia. Human cases were noted mainly in Africa (Zimbabwe, Kenya, Uganda, China) and Asia (India, Indonesia, Iraq), and were caused by the contact and/or alimentary pathways of transmission of the pathogen as a result of forced slaughter of sick and/or butchering of fallen farm animals, consuming meat of sick and fallen cattle. The level of anthrax incidence among farm animals and humans in the Russian Federation in 2022 will be predetermined by the completeness of the implementation of regulated preventive measures, and, provided the strict introduction of comprehensive epizootiological and epidemiological surveillance, will be limited to sporadic cases of infection.

ORIGINAL ARTICLES

71-76 471
Abstract

The aim of the study was to investigate the effect of experimental bases on the growth qualities of liquid nutrient media at the stage of obtaining the biomass of plague microbe vaccine strain using submerged cultivation.

Materials and methods. Yersinia pestis EV NIIEG vaccine strain was used in the work. The cultivation was carried out in a 5 L bioreactor with automatic stirrer control. We used 28 variants of nutrient media obtained through combining five types of bases and six growth stimulants. Nutrient media without the addition of growth-stimulating additives were used as controls. The following parameters were assessed in the yield biomass: the total number of microbial cells, pH, the percentage of viable microbial cells.

Results and discussion. On experimental nutrient media, biomasses of the plague microbe vaccine strain have been produced using submerged cultivation. After evaluating bacterial suspensions by the main indicators, the quality of the obtained suspensions has been compared depending on the nutrient medium used. The most promising bases identified are pancreatic hydrolyzate of casein with dry enzymatic peptone and acid hydrolyzate of corn syrup, especially in combination with such growth stimulants as sodium sulfite, ferrous ammonium sulfate or ammonium molybdate.

77-85 500
Abstract

The aim of the work was to study surface antigenic determinants of V. cholerae R-variant strains using enzyme immunoassay and a panel of monoclonal antibodies (MAbs).

Materials and methods. 60 strains of V. cholerae R-variant isolated from ambient environment objects in the territories of the former USSR and the constituent entities of the Russian Federation over a 30-year period (1988–2019) were investigated in the slide agglutination reaction with cholera diagnostic sera, enzyme immunoassay (ELISA) using the panel of MAbs specific to membrane proteins and a set of reagents “Monoclonal diagnostic immunoglobulins labeled with horseradish peroxidase, dry, for serological identification of V. cholerae O1 and O139 (in vitro) through ELISA and dot-ELISA”.

Results and discussion. The analysis of the surface structures of V. cholerae R-variant strains with atypical agglutinability has been carried out applying enzyme immunoassay. It showed that individual strains with different amounts of O-antigen are registered among the studied strains identified at isolation as V. cholerae R-variant (the optical density range is from 0.261±0.002 to 1.312±0.003). Epitopes of specific O-antigen were found in some “conservative” strains (30 %) that are agglutinated only with RO serum, and in several strains (20 %) that do not have the wbeT gene that determines its synthesis, and lost agglutinability with all diagnostic cholera sera, including RO. The protein epitopes recognized by complementary MAbs are represented with varying frequency in the composition of surface antigens of R-vibrios; a decrease in their representation or absence on the cell surface correlates with the modification or loss of R-LPS and is accompanied by a negative agglutination reaction.

86-93 627
Abstract

The aim of the work was to develop an approach to isolation of rabies virus glycoprotein applying threephase extraction and to characterize its antigenic properties.

Materials and methods. Infectious activity of the rabies virus (production strain, “Ovine” GNKI) after long-term storage was restored on white BALB/c mice. The strain was used for cultivation on BHK-21 cells; the culture liquid was concentrated applying ultracentrifugation followed by separation by buoyant density in a sucrose gradient, selection of visually opalescent zones, phase concentration, chromatographic separation on an ENrich™ SEC650 column (Bio-Rad, USA) and selection of monomeric fractions with high serological activity according to the results of Western blotting.

Results and discussion. We have demonstrated that preliminary mechanical destruction of brain suspension, extraction of the virus-containing material from the cell suspension through successive low-speed and high-speed centrifugation, separation of the sediment produced in sucrose gradient with further phase concentration and chromatographic separation of the precipitate allows to obtain monomeric antigenic preparations with high serological activity. This methodology has made it possible to obtain an antigen, which is rabies virus glycoprotein with a molecular weight of 67 kDa, and two of its isoforms, having molecular weights of 60 and 54 kDa. The described approach can be viewed as an option for isolation of the rabies virus specific antigen when improving laboratory diagnostics techniques. The resulting antigen is a monomeric discrete containing one fraction with a molecu lar weight of 67 kDa. The data obtained corroborate the high specificity of the antigen and its suitability for the design of enzyme immunoassay and immunochromatographic tests, production of specific immunoglobulins, the study of the antigen/antibody interaction, as well as for the assessment of the protective immunity intensity after vaccination.

94-100 890
Abstract

The aim of the research was to study reproduction features of SARS-CoV-2 coronavirus strains of various genetic lines on Vero and Vero E6 cell culture.

Materials and methods. The SARS-CoV-2 virus strains related to the variants of concern (VOC) circulating in the territory of the Russian Federation were used in the research. The strains of the SARS-CoV-2 virus were deposited in the State Collection of Pathogens of Viral Infections and Rickettsioses at the FBIS SSC VB “Vector” of the Rospotrebnadzor. The experiments were carried out on Vero and Vero E6 cell cultures. The dynamics of infectious virus accumulation was determined by titration of culture fluid samples 24, 48, 72, 96 hours after infection (MOI – from 1 to 0,00001 CPE50/cell). Plaque formation was studied on Vero E6 cell culture under 0.2 % agar coating. Image analysis and plaque size calculation were performed using GIMP (GNU Image Manipulation Program).

Results and discussion. The study describes the dynamics of accumulation of infectious virus in the culture fluid depending upon multiplicity of infection for the strains of SARS-CoV-2 virus belonging to different genetic lines. Differences in the morphology of plaques on the monolayer of Vero E6 cell culture under agar coating are shown. SARS-CoV-2 virus strains related to Alfa and Delta VOC demonstrate maximum reproduction rate among the studied strains (infectious titer is higher than 7 lg TCID50/100µl). Omicron VOC forms small plaques under agar coating and at a low multiplicity of infection has a low reproduction rate. Thus, SARS-CoV-2 virus strains belonging to different genetic lines have significant differences in the rate of reproduction on Vero and Vero E6 cell culture.

101-105 393
Abstract

The aim of the study was to assess the epizootiological situation on bacterial vector-borne infections in Caucasian Mineral Waters area of the Stavropol Territory over the period of 2018–2020.

Materials and methods. 3494 specimens of ticks (473 pools), 257 specimens of small mammals, 9 regurgitates of birds of prey and mammals, 7 excreta samples of small mammals, and 2 water samples were tested. Laboratory research of the field material was carried out using molecular-genetic, serological, biological methods. Statistical analysis of laboratory results was conducted using Microsoft Excel 2010. The data were mapped using QGIS 2.18 software.

Results and discussion. The study revealed that the 44.8 % of collected ticks were positive for tick-borne borreliosis, 21.5 % – for tick-borne rickettsiosis, 10.3% – for human granulocytic anaplasmosis, 2.7 % – for Q fever, 0.84 % – for tularemia. There has been an increase in the percentage of positives for tick-borne borreliosis agent samples (more than three times) and a decrease in this indicator for human granulocytic anaplasmosis (1.5 times) as compared with 2010–2012. Investigation of tick infection with the agents of Q fever and tick-borne rickettsioses has not been previously conducted in the region. During the period under review, 19 pools of ticks had mixed infection, which indicates that there are combined foci of bacterial natural-focal infections with vector-borne transmission in the recreation zone of the Stavropol Territory. This necessitates preventive measures and systematical epizootiological surveys in the Caucasian Mineral Waters region.

106-112 371
Abstract

The aim of the study was to analyze differential gene expression in Burkholderia pseudomallei strains with different survival rates under cold stress conditions.

Materials and methods. Three strains of B. pseudomallei (sequence types ST 46, ST 70, and ST 85) were used as model strains. The RNA was isolated using the membrane columns method and stabilized through dehydration. The cDNA was sequenced on the Illumina MiSeq platform. Gene functions were classified using the KEGG PATHWAY database.

Results and discussion. Based on the analysis of transcriptomes of B. pseudomallei strains after prolonged exposure to cold stress, the molecular mechanisms of B. pseudomallei adaptation to low temperatures have been described for the first time ever. It was shown that adaptation of B. pseudomallei to cold stress is associated with regulatory processes leading to a significant decrease in the total transcriptional activity. Two strategies of adaptation to low temperatures have been found: 1) modulation of regulatory processes leading to suppression of gene expression of the main metabolic pathways to the minimum level that ensures cell viability and activation of the minimum required set of stress response genes, and 2) less noticeable suppression of general metabolism in combination with activation of expression of an extended range of genes for cold and heat shock, general, osmotic, and universal stresses. Both mechanisms provide the causative agent of melioidosis with survival under conditions of prolonged cold stress at low positive temperatures. The first strategy showed greater efficiency at negative temperatures. The transition of B. pseudomallei to a viable but uncultivated state occurs in the long term (at least 2 years). While with the second strategy, this happens within 2 months. Assessment of the potential and molecular mechanisms of adaptation of this bacterium to cold stress is necessary to understand the degree of risk in case of a possible introduction of B. pseudomallei into regions with a temperate climate and to develop effective measures to ensure the biosafety of the environment.

113-121 550
Abstract

The aim of the study was to assess the dynamics of seroprevalence to SARS-CoV-2 in various population groups during the initial period of herd immunity formation based on multivariate analysis using logit regression models.

Materials and methods. The study involved 1561 individuals divided into three population cohorts: people living with HIV/AIDS (PLHA), healthy blood donors, medical workers. The presence of antibodies to SARS-CoV-2 was determined in blood serum through ELISA using commercial reagent kits. Multivariate analysis of the dynamics of seroprevalence to SARS-CoV-2 was carried out using logistic regression models.

Results and discussion. It has been revealed that probability of detecting IgG antibodies to SARS-CoV-2 significantly increased among the donors and medical workers in the spring-autumn 2020 series (p=0.005 и p<0.001, respectively), which corresponds to seroprevalence shift in the general population. Groups of donors and medical workers can be considered as indicator groups that characterize the herd immunity in reference to SARS-CoV-2, as well as the intensity of COVID-19 epidemic process. Seroprevalence to SARS-CoV-2 in PLHA group was at a consistently high level throughout the observation period. The generated logistic regression models made it possible to determine the trends in the development of the epidemic situation based on multifactorial analysis of the dynamics of seroprevalence to SARS-CoV-2.

122-129 560
Abstract

The aim of the work was to conduct phylogenetic analysis of Y. pestis strains, biovar altaica ssp. central asiatica, isolated in Gorno-Altai high-mountain and Sailyugem natural plague foci on the territory of Russia and Mongolia in 1965–2020, by full-genome sequencing data.

Materials and methods. To determine the population structure of the biovar altaica ssp. central asiatica, 34 whole genome sequences were used (including 20 Y. pestis strains of the biovar altaica, 18 of which were sequenced by us). To isolate DNA from the Y. pestis strains, a PureLink Genomic DNA Mini Kit (Invitrogen, USA) was applied. Sequencing of the nucleotide sequences of Y. pestis strains was carried out in Ion PGM system Lifetechnologies. The analysis and processing of the obtained data were performed with the help of Newblergs Assembler 2.6 and IonTorrent Suite software package, 3.4.2. The search for SNPs was performed using the Wombac 2.0 program. The Maximum Likelihood dendrogram was built in the PhyML 3.1. The dendrogram was visualized using the FigTree 1.4.3 software.

Results and discussion. Based on the data of whole genome analysis, taking into account the 1871 revealed polymorphic nucleotides, the spatial structure of the biovar altaica ssp. central asiatica has been determined. It includes several phylogeographic branches: the Kurai-Tarkhatinskaya (cluster 0.PE4a-1) and the Ulandryk-Mongolian (0.PE4a-2), which is in agreement with the geographical regions of the isolation of strains forming these branches in the Altai Mountains. The Kurai-Tarkhatinskaya branch is further divided into the Kurai (sub-cluster 0.PE4a-1-1, formed by the strains of 2009–2018) and Tarkhatinskaya (subcluster 0.PE4a-1-2, formed by the strains of 2012–2020) sub-branches, while the Ulandryk-Mongolian branch of evolution is split into sub-branches represented by strains from the Ulandryk meso focus (sub-cluster 0.PE4a-2-2, strains 1965–2010) and the Sailyugem focus of Mongolia (sub-cluster 0.PE4a-2-1, strains 1964–1990).

130-136 464
Abstract

The aim of the work was to summarize the experience of the Rostov-on-Don Research Anti-Plague Institute in the organizational and functional response aimed at ensuring the implementation of a complex of on-going and scheduled anti-epidemic (preventive) measures at different territorial levels in the context of COVID-19 pandemic. Stage-wise reassignment of scientific and operational subdivisions of the facility was carried out in a timely manner in a flexible mode for a prompt response to the challenges that arose from community transmission of the new coronavirus infection in the territory of the Russian Federation. It allowed an effective distribution of the forces and assets without damage for specialized functioning in the main areas of scientific and practical activities. Realized was an effective interdepartmental interaction with territorial scientific and practical institutions of the Rospotrebnadzor and specialized medical organizations. Practical, methodological and advisory assistance was provided to personnel of specialized medical institutions within the framework of conversing in-patient facilities into hospitals for the treatment of patients with COVID-19. Priority research to study the level of the humoral and cellular immune response to SARS-CoV-2 among the population of the Rostov Region, the etiological spectrum of the causative agents of community-acquired pneumonia associated with COVID-19, full genome sequencing of SARS-CoV-2 isolates was carried out. Specialists of the task forces reinforced human resources of individual laboratory facilities in Moscow city, the Republic of Crimea and the Rostov Region. Complex objective of effective functioning has been implemented: in an operational mode – participation in anti-epidemic (preventive) measures in the face of changes in the dynamics and intensity of the epidemic manifestations of new coronavirus infection; in scheduled mode – the implementation of activities in the main areas of the scientific work of the organization.

137-147 422
Abstract

The aim of the work was to study the prevalence of different types of SXT element with different composition of antibiotic resistance genes among clinical strains of the El Tor cholera pathogen isolated in Russia, Ukraine and cholera-endemic countries in Asia and Africa.

Materials and methods. The subject of the study was 27 strains and nucleotide sequences of 77 strains of Vibrio cholerae El Tor available from the NCBI GenBank. The structure of the SXT element and its type were determined using the Mauve and BLAST v.2.9.0 programs. Phylogenetic relations of strains with different types of SXT were identified using Snippy v.4.6.0 and MrBayes v.3.2.7 software. Assessment of strain sensitivity to antibiotics was carried out in accordance with Methodological Regulations 4.2.2495-09.

Results and discussion. Two types of SXT element (ICEVchInd5 and ICEVchBan9) have been identified among the studied strains from Russia and Ukraine, which have different composition of antibiotic resistance genes: floR, strAB, sul2, dfrA1 and floR, tetAR, strAB, sul2, dfrA1, respectively. At the same time, the studied strains from Asia and Africa contain five types of SXT: ICEVchInd5, ICEVchBan9, ICEVchBan5, SXTTET, ICEVchInd5ΔVRIII, which differ in size and/or composition of resistance genes. Of these, the last three have not been found in Russia and Ukraine. Due to the high level of genomic diversity of SXT in the population of V. cholerae in endemic regions, there is a risk of importation of cholera pathogen strains with altered resistance to antibiotics into Russia. Phylogenetic relations of 76 strains with different SXT types and different alleles of the ctxB gene encoding the B subunit of cholera toxin have been assessed based on SNP analysis. A close phylogenetic relation between strains with the same type of SXT isolated in Russia and Asian countries has been demonstrated, which confirms the importation of the causative agent of cholera with multiple resistance to antibiotics from this region and the need for constant monitoring of the sensitivity of V. cholerae to antimicrobial drugs.

148-155 681
Abstract

The aim of the research was to assess the susceptibility of mice of different lines to newly emerging variants of SARS-CoV-2.

Materials and methods. The SARS-CoV-2 virus strains belonging to variants of concern (VOC) circulating in the territory of the Russian Federation were used in the study. Experiments involved three inbred mouse lines (BALB/c, CBA and C57Bl/6z) and CD1 outbred mice taken from the nursery of the SSC VB “Vector” of the Rospotrebnadzor. The infectious titer of coronavirus in tissue samples obtained from the laboratory animals was determined on a Vero E6 cell culture. The (Ct) threshold value in RT-PCR was considered an additional parameter for monitoring the viral load in the samples. The severity of lung tissue damage was assessed using histological preparations.

Results and discussion. The susceptibility of various mouse lines to the genetic variant Beta of the SARS-CoV-2 virus has been investigated. During intranasal infection of the inbred and outbred mice with strains of VOC at a dose of 2·103 TCID50, the virus replicated in the lungs with maximum concentrations 72 hours after infection. The pathogenicity of genetic variants of the SARS-CoV-2 virus for BALB/c mice has been assessed, a 50 % infectious dose for intranasal infection (ID50) determined. Histological analysis showed COVID-19-specific lung tissue lesions in infected animals. Our study proves that BALB/c mice can be used as a model animal in screening studies when evaluating the effectiveness of therapeutic, vaccine preparations and studying the pathogenesis caused by VOC of the SARS-CoV-2 virus: Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Omicron (B.1.1.529) and the like.

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ISSN 0370-1069 (Print)
ISSN 2658-719X (Online)