REVIEWS
Natural foci of plague – a particularly dangerous transmissible zoonotic disease – are widely spread across the world, including in the territory of Russia and a number of neighboring countries in Central Asia. Between 2000 and 2019, 28339 cases of plague were registered in 21 states around the world; lethality rates reached 8 %. Active plague manifestations are associated with the increase in the abundance of carriers and vectors of the disease, which increases the risk of human infection. Prophylactic measures against plague are aimed at prevention and eradication of epidemic foci; they incorporate a whole complex (system) of activities. Disinsection and deratization remain the most radical of them. Control of the numbers of carriers and vectors of plague involves decrement up to the levels at which epizooties will not develop, they are suppressed and liquidated. Currently in the Russian Federation and CIS countries, the focus is on the field and community disinsection when planning and performing exterminatory interventions. The decrease in the numbers of fleas – carriers and vectors of plague microbe – results in disruption of epizootic process and, consequently, in provision of epidemiological welfare as regards this dangerous disease. Insecticide treatments in natural biotopes are carried out using powder-like or aerosolized products, the class of pyrethroids, organophosphorous compounds and phenylpyrazoles by means of advanced equipment: knapsack dusters, power operated air blowers, and foggers. Within the boundaries of residential areas wet disinsection is performed with the help of spraying devices. Field deratization is allowed upon epidemiological grounds only, in localized territories of epizooties, within the boundaries of the settlements of plague infected carriers. Zinc phosphide- or anticoagulant-based bait products are used as rodenticides. Therewith, the eradication of valuable animal species, rare species of small mammals and endemic species is prohibited. Community deratization is always accompanied by the community disinsection. When planning and carrying out insecticide and rodenticide treatments the priority is given to the safety of handling the chemical preparations, to the exclusion of the negative effect on target animal species.
The endemicity of cholera in India has been well researched. Among the other endemic areas, Indian subcontinent appears to be the cradle of Vibrio cholerae genovariants, which subsequently spread worldwide. In contrast, all the cholera cases recorded in Russia are of imported origin. In the past century, such importations might result in epidemics, which, however, ended with elimination of toxigenic V. cholerae (TVC) from the affected areas. Currently, the incidence of TVC in water reservoirs or infected returnees from Asian countries are rare events, mostly due to constant surveillance activities. Furthermore, the climatic conditions in the majority of Russian regions are unfavorable for longterm environmental survival of the pathogen. On the other hand, global shifts in climate accompanied by unexpected anomalies in previously stable climatic zones may promote dissemination of imported TVC and emergence of cholera. In some regions of Russia, seasonal weather patterns are pretty similar to Indian. Molecular studies of isolates from diverse territories show that TVC have been introduced into new areas and acquired additional traits, absent in their ancestors.
This article describes genomic characteristics of TVC isolates from Russia, India and some other countries. The review is complemented by bioinformatics analysis of important genetic markers to discriminate between clones that emerged in endemic regions and were imported into cholera-free locations. We have found more evidence in support of primary Indian origin of recent genovariants and their ongoing transformation, including the emergence of a new rtxA gene allele with a 60 bp deletion in addition to the previously known null mutation. In certain conditions, TVC could pose a potential threat of spreading epidemic cholera into Russia and other territories. Cholera control requires joint efforts of researchers to learn more about the pathogen at the molecular level for developing effective strategies to protect the humankind throughout the world.
ORIGINAL ARTICLES
The ongoing COVID-19 pandemic around the world and in Russia remains a major event of 2020. All over the world, research is being conducted to comprehensively study the patterns and manifestations of the epidemic process. The main quantitative characteristics of SARS-CoV-2 transmission dynamics among the population, based on the data of official monitoring over the current situation, play an important role in the development of the epidemiological surveillance system.
The aim of this study is to explore the peculiarities of age-gender distribution of COVID-19 patients in Moscow.
Material and methods. The data related to the epidemiological characteristics of age-gender structure of COVID-19 patients in Moscow between March 19, 2020 and April 15, 2020, at different stages of the epidemic were retrospectively analyzed.
Results and discussion. The mean age of COVID-19 patients in Moscow was 46,41±20,58 years. The gender ratio (male/female) among the patients was 52.7/47.3 %, wherein the indicators varied depending upon the age. Male/female ratio in the age group “under 39” stood at 53.7/46.3 %, and “over 40 years of age” – at 39.3/60.7 %. The predominant age range among male cases was 19 to 39 years old – 35.4 %, while among female patients – 40–59 years (36.5 %). The age distribution of patients in Moscow is indicative of the fact that COVID-19 is a disease that primarily affects older age groups. The age structure of all COVID-19 cases during the observation period is characterized by predominance of adult patients over 19 years of age – 92,7 % (92,6–92,8 %), the share of patients aged 40–59 years is 35,7% (35,5–35,9 %). The differences in the age distribution in males and females are as follows: in the male cohort, the age groups 19–39 years old and 40–59 years old prevail – 35.4 % (35.1–35.7 %) and 34.9 % (34.6–35.2 %), respectively. The age group 40–59 years old – 36.5 % (36.3–36.8%) dominates in the female cohort.
Aim. To study the spread of COVID-19 among the population of the Omsk Region during 24 weeks of the epidemic on the background of anti-epidemic measures.
Materials and methods. A descriptive epidemiological study was carried out based on publically available data и data from the Center for Hygiene and Epidemiology in the Omsk Region on the official registration and epidemiological investigation of detected COVID-19 cases in the Omsk Region for the period from March 27 to September 10, 2020. To assess the potential of COVID-19 to spread, the following indicators were calculated: exponential growth rate (r), basic reproduction number (R0), effective reproduction number (Rt), expected natural epidemic size and herd immunity threshold. Data processing was performed using MS Excel 2010. The cartogram was built using the QGIS 3.12-Bukuresti application in the EPSG: 3576 coordinate system.
Results and discussion. For the period from March 27 to September 10, 2020, a total of 9779 cases of COVID-19 were registered in the Omsk Region, the cumulative incidence was 507,6 per 100000 (95 % CI 497,5÷517,6), the case-fatality rate for completed cases was 2.9 %, for identified cases – 2.4 %. The most active spread of COVID-19 was noted in Omsk and 4 out of 32 districts of the region (Moskalensky, Azov German National, Mariyanovsky, Novovarshavsky). During the ongoing anti-epidemic measures, the exponential growth rate of the cumulative number of COVID-19 cases was 4.5 % per day, R0 – 1.4–1.5, Rt – 1.10, herd immunity threshold – 28.6 %. The expected size of the epidemic in case of sustained anti-epidemic measures can reach 58.0 % of the recovered population. A decrease in the number of detected virus carriers, incomplete detection of COVID-19 among patients with community-acquired pneumonia introduced additional risks for the latent spread of infection and complications of the epidemic situation. Maintaining restrictive measures and increasing the proportion of the immune population (over 28.6 %) may significantly reduce the risks of increasing the spread of COVID-19 in the Omsk Region.
Objective: to study the bacterial microflora in the sputum of patients with pneumonia caused by SARSCoV-2 or other pathogens.
Materials and methods. The bacterial microflora of sputum of 173 patients with pneumonia admitted to hospitals in Khabarovsk and the Khabarovsk Territory in May – June 2020 was examined. Detection of RNA of the SARS-CoV-2 virus was carried out by PCR with the Vector-PCRRV-2019-nCoV-RG test system (manufactured by the State Scientific Center of VB “Vector”, Koltsovo). Determination of the DNA of mycoplasmosis agents and chlamydia was carried out with the test system “AmpliSens® Mycoplasma pneumoniae/Chlamydophila pneumonia” (manufactured by CRIE). Statistical data processing was performed using the Excel program.
Results and discussion. Both groups of patients (Covid-19+ and Covid-19–) had high levels of bacterial flora isolation (81.4 and 74.7 %) including common pathogens for community-acquired pneumonia as well as notable detection frequency of Candida spp. and microbial associations. The group of Covid-19+ patients demonstrated a wider range of detected pathogens, was positive for poly-resistant gram-negative Enterobacteriaceae, non-fermenting gram-negative poly-resistant bacteria, with more expressed manifestation of microbial associations. In the group of Covid-19– participants drug-resistant microflora was presented only by MRSA and MRSE staphylococci.
Objective: to study the possibility of applying electro-optical analysis for the assessment of cell viability of tularemia microbe vaccine strain at different stages of experimental live tularemia vaccine production.
Materials and methods. The research object was a cell culture of Francisella tularensis 15 NIIEG. Investigations were carried out at all stages of experimental live tularemia vaccine (ELTV) manufacturing according to an advanced technology: cultivation, concentrating, diafiltration, mixing with drying media, stabilization, and storage (two-year period of observation). Electro-optical analysis by the parameter “polarizability anisotropy” of bacterial cell was conducted with the help of EloTrace (EloSystems, Germany). Total concentration of cells was evaluated using density metering at 590 nm and spectrometry – at 650 nm. Viability was assessed through inoculation of plates with FT-agar.
Results and discussion. The experiment has demonstrated that the change in polarizability anisotropy of the cell at the frequencies of 900 kHz and 2100 kHz, reflecting the state of cytoplasm and cytoplasmic membrane, respectively, is the earliest response to changes in vital indicators of bacterial culture in the process of cultivation. Thereby, the decrease in viability of F. tularensis cells occurrs well before the decrease in cell concentration. We have shown the preservation of viability of F. tularensis 15 NIIEG cells at all stages of experimental live tularemia vaccine production. Electro-optical analysis allows for registering the changes in vital parameters of microorganism cells in real-time mode, while the assessment of viability applying bacteriological method takes up to 5 days. Different stages of tularemia vaccine manufacturing have impact on the vital indicators of F. tularensis cells, and electro-optical analysis is a prospect method of control of such parameter as “Specific activity (the number of live microbial cells)”.
Objective of the study was to analyze phylogenetic relation of the strains and dissemination routes of Yersinia pestis in Kara Kum desert focus in XX century.
Materials and methods. We have carried out whole genome sequencing of 8 Yersinia pestis strains isolated in Kara Kum desert focus between 1949 and 1985. We also used nucleotide sequences of 16 Y. pestis strains from neighboring foci of East and North Caspian Sea region dated 1917–2002, sequenced at the premises of RusRAPI “Microbe” on Genestudio Ion S5XL platform (Thermo Fischer Scientific). Phylogenetic analysis was performed based on 1720 identified core SNPs. Phylogenetic dendrogram was constructed using Maximum Likelihood algorithm, PhyML v. 3.1 software package, and nucleotide substitution model GTR.
Results and discussion. On the basis of whole-genome SNP analysis of Y. pestis strains from Kara Kum desert focus, West and North Caspian Sea region foci analysis of plague agent populations circulating in Turkmenistan in XX century was conducted. It has been demonstrated that three separate waves of Y. pestis dissemination swept the territory of Kara Kum desert focus. The origin of the first one, registered in 1912, plague outbreak in south-eastern part of the focus and its etiological agent remain unspecified so far. The second wave is associated with distribution of North-Aral 2.MED1 population of medieval biovar in 1949–1970, which led to the emergence of epizootic activity in 1949, as well as subsequent cases and outbreaks of plague in this territory in the middle of the XX century. In the mid-second half of the past century, one more 2. MED1 dissemination wave from the North Balkhash region hit the Kara Kum desert focus. This central-asian population has also taken root in the natural biocoenosis of the Kara Kum focus. The data obtained testify to active processes of distribution of Y. pestis of medieval biovar in East Caspian Sea region in southern sub-zone of Eurasian deserts in XX century.
Francisella tularensis is an intracellular bacterium that causes tularemia. Progress in creating a safe and effective vaccine for the prevention of tularemia is challenging due to a lack of knowledge about immunological parameters indicative of protective adaptive immunity. Objective of the research was to assess the effect of modifications of the F. tularensis 15 NIIEG genome on the immunogenic and protective properties of F. tularensis 15/23-1ΔrecA and F. tularensis 15/23-1/sodBΔrecA strains. Materials and methods. Multi-parameter flow cytometry and the measurement of secreted cytokines were used to characterize the responses of mouse spleen lymphocytes in response to re-stimulation of F. tularensis with acid-insoluble complex (AIC) in vitro. Also, the titers of specific antibodies to F. tularensis lipopolysaccharide in blood serum were analyzed by enzyme-linked immunosorbent assay. Results and discussion. It has been shown that immunization with the studied strains led to a significant increase in CD4+ and/or CD8+ T cells capable of expressing functional markers: CD69, CD25 and/or CD28; an increase in the subpopulation of T-helpers synthesizing IFN-γ. In the body of immune mice, a pool of B-lymphocytes was formed, capable of secreting IFN-γ in response to their stimulation with AIC. Immunization with the strain 15/23-1/sodBΔrecA provided 70% protection in mice from intranasal infection with a virulent strain of F. tularensis SchuS4. More pronounced protective properties were associated with the activation of not only B-lymphocytes and T-helpers, but also with the simultaneous activation of cytotoxic T-lymphocytes.
Objective: to study the evolution of SARS-CoV-2 virus in the process of adapting to human organism during
the current pandemic.
Materials and methods. Database (GISAID) on nucleotide sequences of SARS-CoV-2 virus genome, obtained from clinical samples during the period of late December, 2019–July, 2020. Phylogenetic tree diagram construction was carried out applying BioNumerics v.7.6 software using Maximum parsimony algorithm.
Results and discussion. The most substantial change in the genomes of SARS-CoV-2 virus are associated one-time mutations in ORF1b (P314L) and S (D614G) genes, as a result of which the overwhelming majority of identified isolates of this virus have the stated pair of substitutions to date. Many researches link the substitution in S (D614G) gene to the decreased pathogenicity in the strains that contain it, which may be also explained by enhanced methodology of patient treatment in the course of pandemic. The effect of the mutation in ORF1b (P314L) gene has not yet been investigated. P314L and D614G mutations are closely related and only their combined presence in the genome favored the dissemination of the genovariants of SARS-CoV-2 virus. Analysis of congregated epidemiological data testifies to the fact that the spread of new genovariants may be associated with biological properties facilitating human-to-human transmission. Thereat, associated decrease in lethality may reflect not only advancements in methods of treatment, but possible attenuation of virulent properties. Thus, observed growth in dissemination potential against the background of decrease in virulence is, probably, one of the forms of adaptation of new coronavirus to human population and, apparently, will remain in the future as the integration of SARS-CoV-2 virus into the structure of seasonal ARVI agents.
In order to develop the most diagnostically informative methods for carrying out antigen-stimulated cellular tests in vitro a careful selection of stimulating agent (antigen) is required, possessing an adequate activating potential and providing specificity of the reaction.
Objective of the study was to identify the qualitative indicators of experimental batches of brucellosis antigen preparations designed for cellular tests in vitro.
Materials and methods. Initially we produced antigen complexes of brucellosis microbe on the basis of the vaccine strains of three epidemically significant Brucella species (B. abortus, B. melitensis, B. suis). Quantitative determination of WsAg and PPBC proteins of experimental preparation series was performed applying capillary electrophoresis. Qualitative composition was assessed through ion exchange liquid chromatography with refractometric detection.
Results and discussion. We have specified physical-chemical features, investigated chromatographic profiles and composition of protein fractions, as well as tried the produced experimental batches of brucellosis antigen preparations. After analyzing the defined protein and polysaccharide composition of the obtained WsAg samples, one can conclude that WsAg preparation cannot be used for cellular tests as the probability of non-specific lymphocyte reaction manifestation in vitro was experimentally proven. By contrast, complex brucellosis antigen preparation PPBC has an expressed specific activity and specificity under in vitro conditions and the prospects to be used when developing methodological approaches for laboratory diagnosis of brucellosis and assessment of de facto immunity rate in risk contingents after vaccination. The obtained parameters will allow for proper quality provision when manufacturing the developed experimental PPBC preparation designed for cellular tests in vitro, taking into account modern validation and standardization regulations.
Aim. Analysis of the phenotypic characteristics and identification of peculiarities of the genotypic organization in non-toxigenic strains of cholera vibrios having different origin, isolated in Russia.
Materials and methods. A sample of 548 non-toxigenic strains obtained using the author’s updated GIS “Cholera 1989–2014” was used. PCR genotyping was carried out in accordance with the patented “Method for the identification of non-toxigenic strains of cholera vibrio O1 serogroup using PCR to isolate genetic determinants.” Cluster analysis was performed applying the UPGMA method. The dendrogram was constructed using MEGA 5 software package.
Results and discussion. Representative cultural-morphological, serological and biochemical properties of V. cholerae strains have been specified. The variability of the studied strains on the basis of phagolizability has been revealed. Unique phage-types not previously encountered in Russia have been identified. The population of non-toxigenic strains of cholera vibrio O139 serogroup is genetically homogeneous in contrast to V. cholerae O1 El Tor isolates and has identical PCR genotypes. The universality of the PCR genotyping by 14 target genes has been shown to differentiate the studied strains of V. cholerae O1 and O139, as well as to identify disparities among O139 strains isolated in different geographical regions of the country.
Objective of our work was to assess prevalence of the primary HCV drug resistance mutations in the NS5b gene in patients with newly diagnosed HIV infection.
Materials and methods. The study material was 196 blood plasma samples from patients living in the North-Western Federal District with newly diagnosed HIV. Samples were examined for the anti-HCV antibodies and HCV RNA presence. If HCV RNA was detected, amplification was performed using three primers pairs that co-flanked the NS5b gene. After sequencing the indicated gene nucleotide sequence, the virus subtype was determined and drug resistance mutations were detected.
Results and discussion. Antibodies to HCV were detected in 18.87 % of HIV-infected individuals. HCV RNA was detected in 18.36 % of the patients, including 89.18 % anti-HCV-positive and 1.88 % anti-HCV-negative. It was shown that co-infection is more common in men (77.8 %) compared to women (22.2 %) – χ2 = 3.996 at p = 0.0456, df = 2. The difference in the HIV viral load between the groups with HIV monoinfection and with HIV + HCV coinfection was demonstrated (χ2 = 6.284 at p = 0.0432, df = 2). A significant difference between the groups by the CD4 + lyphocytes number was shown. In the phylogenetic analysis, the HCV subtypes are distributed as follows: HCV 1b – 47.2 %, HCV 3a – 30.6 %, HCV 1a – 13.9 %, HCV 2a – 5.5 % and only one sample was defined as HCV 2k – 2.8 %, respectively. Nine samples (25 %) presented NS5b mutations in the positions related to the development of drug resistance of HCV, including two samples among HCV genotypes 1a and 3a (i.e., 5.6 % of the total HIV + HCV group), as well as five samples among HCV 1b (13.9 % of the total group). Mutations among HCV 1a were C316Y and N444D substitutions. Among HCV 1b, C316N, C451S, S556N/G substitutions were identified. Among patients with HCV 3a, 2 samples (5.6 %) with a D310N mutation associated with an unfavorable disease prognosis were found. The introduction of direct sequencing of HCV nucleotide sequences into the routine laboratory diagnostics will allow us to estimate the primary drug resistance mutations prevalence in risk groups to predict the HCV life-threatening complications development – fibrosis, cirrhosis, hepatocellular carcinoma, as well as the outcome of antiviral therapy prognosis. The data obtained can be rationally used to assess the dynamics of the HCV primary pharmacoresistance prevalence among HIV-infected individuals.
Objective of the seroepidemiological study was to determine the level and structure of herd immunity to SARS-CoV-2 among the population of the Irkutsk Region during the period of an increase in the incidence of COVID-19.
Materials and methods. The content of antibodies to SARS-CoV-2 was determined by ELISA applying a reagent panel “ELISA anti-SARS-Cov-2 IgG” produced by the State Scientific Center of Applied Microbiology and Biotechnology (Obolensk).
Results and discussion. The investigation has revealed that the herd immunity of the total population of Irkutsk Region amounted to 5.8 %. The greatest share of seropositive persons was among the children aged 14–17 (13.8 %) and 1–6 (11.8 %). It has been established that the risk of infection increases by 3.1 times in case of contact with COVID-19 patients. After exposure to COVID-19, antibodies were produced in 56.5 % of the cases. The share of asymptomatic forms among seropositive residents of the Irkutsk Region reached 81.2 %. Consequently, during the chosen period of increased COVID-19 incidence among the population of Irkutsk Region low level of seroprevalence was formed. A significant proportion of asymptomatic forms of infection characterize high intensity of the latently developing epidemic process. The results obtained can be used when organizing preventive measures, including vaccination, and for forecasting morbidity rates.
The first case of COVID-19 was registered in the Leningrad Region on March 13, 2020. The period of increasing intensity of the epidemic process lasted 8 weeks. One month after reaching the maximum incidence rate, a study was organized to determine seroprevalence to COVID-19 among the population of the Region.
Objective of the study was to determine the level and structure of community immunity to SARS-CoV-2 virus in the population of the Leningrad Region in the period of intensive COVID-19 transmission.
Materials and methods. The work was carried out within the framework of the Rospotrebnadzor project on assessment of community immunity to the SARS-CoV-2 virus in the population of the Russian Federation. The content of antibodies to SARS-CoV-2 was determined applying ELISA using a kit for the analysis of human serum or blood plasma for the presence of specific IgG to the nucleocapsid of the SARS-CoV-2 virus , manufactured by the State Scientific Center of Applied Microbiology and Biotechnology (Obolensk) in accordance with the instructions for use.
Results and discussion. The study has showed that the herd immunity of the population of the Leningrad Region was 20.7 %. The maximum level has been established in children 1–6 years old (42.3 %) and people over 70 years old (29.0 %). The highest level of seropositivity, except for children and older people, was found among the unemployed (25.1 %). The lowest level of seroprevalence was detected in civil servants (12.8 %) and military personnel (16.7 %). It has been shown that the risk of infection increases by 1.5 times in case of contacts with COVID-19 patients. After exposure to COVID-19 virus, antibodies are produced in 82.1 % of the cases. In individuals with a positive PCR test result obtained earlier, antibodies are detected in 82.8 % of cases. The share of asymptomatic forms among seropositive residents of the Leningrad Region was 86.9 %. The results of assessing the herd immunity to SARS-CoV-2 in the population of the Leningrad Region indicate that during the period of intensive COVID-19 transmission, an average level of seroprevalence was formed. A significant proportion of asymptomatic forms of infection characterizes the high intensity of the latently developing epidemic process. The results obtained should be taken into account when organizing preventive measures, including vaccination, and predicting morbidity rates.
The first case of COVID-19 was diagnosed in St. Petersburg on March 2, 2020; the period of increase in the incidence lasted for 10 weeks, the maximum rates were recorded in mid-May, and subsequently there was a statistically significant decrease in the incidence.
Objective: to determine the level and structure of community immunity to SARS-CoV-2 among the population of St. Petersburg during the period of intensive spread of COVID-19.
Materials and methods. Selection of volunteers for the study was carried out through interviewing and randomization. The exclusion criterion was active COVID-19 infection at the time of the survey. 2713 people aged 1 to 70 years and above were examined for the presence of specific antibodies to SARS-CoV-2. Antibodies were detected by enzyme immunoassay.
Results and discussion. Studies have shown that in St. Petersburg, in the active phase of COVID-19 epidemic, there was a moderate seroprevalence to SARS-CoV-2, which amounted to 26 %, against the background of a high frequency (84.5 %) of asymptomatic infection in seropositive individuals who did not have a history of COVID-19 disease, positive PCR result and ARI symptoms on the day of examination. The maximum indicators of herd immunity were established in children 1–6 years old (31.1 %), 7–13 years old (37.7 %) and people over 70 years old (30.4 %). Differences in the level of seroprevalence in the age groups of 18–49 years are statistically significant. The highest level of seroprevalence was found among the unemployed (29.7 %), healthcare workers (27.1 %), education sector (26.4 %) and business sector personnel (25 %). In convalescents, COVID-19 antibodies are produced in 75 % of cases. In individuals with positive result of PCR analysis carried out earlier, antibodies are detected in 70 % of the cases. The results of the study of herd immunity to SARS-CoV-2 are essential to forecast the development of the epidemiological situation, as well as to plan measures for specific and non-specific prevention of COVID-19.
Objective: to analyze the epidemiological situation on Ixodidae tick-borne borreliosis in the Russian Federation in 2019 in comparison with the period of 2002–2018.
Materials and methods. The paper uses the data contained in Form No. 2 of the state statistical reporting for 2002–2019 and information obtained by the Reference Center for Monitoring Borreliosis of the Omsk Research Institute of Natural Focal Infections from 74 constituent entities of Russia in 2019. The main research method is epidemiological one with the use of modern information technologies.
Results and discussion. In Russia, 8048 cases of tick-borne borreliosis (Lyme disease – LD) were recorded (5.48 0/0000) in 2019. The actual indicators of the LD incidence for the whole country and federal districts (FD) in 2019 were within the confidence limits predicted with linear regression based on the study of the dynamics of the epidemic process in 2002–2018 in the vast majority of cases. A steady upward trend in the LD incidence was observed during 2002–2019 in the Central Federal District due to 10 out of 18 entities (Moscow, Belgorod, Lipetsk, Moscow, Bryansk, Voronezh, Kursk, Ryazan, Tambov, Tula regions); in the Southern Federal District because of the Krasnodar Territory and the Volgograd Region; in the North Caucasus Federal District – because of the Stavropol Territory. Despite the fact that a downward trend in the incidence of LD has been established over the past 18 years in the North-West, Volga and Ural Federal Districts, in some subjects of these regions a trend towards an aggravation of the epidemiological situation is observed (the Komi Republic and Chuvashia, Penza Region). In the absence of a pronounced tendency to change in the incidence rate of LD in the Siberian and Far Eastern Federal Districts, Kemerovo Region-Kuzbass, the Republic of Tuva and the Trans-Baikal Territory where a growing trend has been identified require special attention. In the Ural and Siberian Federal Districts, the share of non-erythema forms among laboratory-confirmed cases of LD was higher than in other regions, which merits further study of the genome-specific features of borrelia populations and their carriers. Effective control of the LD epidemiological situation in Russia is possible provided that the control is improved and maintained, and the capacity of preventive measures and zoological-entomological monitoring of the activity and structure of the natural foci of LD is enhanced in the entities with the long-term tendency towards increase in the incidence of the disease.
Objective of the study was to select the criteria and methodological approaches to reclassify avirulent strains of plague agent from pathogenicity (hazard) group I into pathogenicity group III.
Materials and methods. We have reviewed domestic and foreign normative, methodological documents, scientific publications in the field of laboratory diagnostics of plague and biosafety provision while working with pathogenic biological agents.
Results and discussion. A complex of criteria for reclassification of Y. pestis strains from hazard group I into hazard group III has been substantiated; the methods for their assessment identified. Validation has revealed the grounds for reassignment of a number of avirulent Y. pestis strains included into the training kit which is compiled for mastering the training module “Microbiology and laboratory diagnostics of plague” into pathogenicity group III. We have demonstrated the feasibility of combining the structured methods of assessment of pathogenic properties in plague microbe with additional informative ones; in particular, evaluation of cytotoxicity of Y. pestis strains in relation to leucocytes of whole human blood in vitro. Analysis of a strain virulence should be built on complex characterization of major pathogenicity factors using advanced molecular-genetic research methods, the data on phenotypic manifestations of their functioning, as well as the level of pathogenicity for sensitive laboratory animals. The review of the utilized quantitative indicators for differentiation of plague microbe strains by virulence taking into account LD50 values for Y. pestis EV NIIEG strain is a relevant task. It is practical to supplement the complex approach with informative research methods, notably, characterization of strain cytotoxicity which shows high correlation with virulence criterion LD50.BRIEF COMMUNICATIONS
The aim of this work was to identify the causative agent of community-acquired pneumonia and coinfection using PCR study of biomaterial from patients.
Materials and methods. PCR testing of 268 samples from 258 patients was carried out to identify RNA/DNA of viral and bacterial pathogens of respiratory infections.
Results and discussion. In 43.3 % of samples SARS-CoV-2 RNA was detected, in 4.5 % – RNA/DNA of acute respiratory viral infections pathogens, in one sample – DNA of Mycoplasma pneumoniae. Co-infection was detected only in patients of the anti-tuberculosis dispensary (SARS-CoV-2 and Mycobacterium tuberculosis). In the examined patients with pneumonia, SARS-CoV-2 RNA was significantly more often detected in biomaterial from the lower respiratory tract (52 %) than in respiratory smears (8.5 %). In the first week from the onset of the disease, 19.2 % of positive samples were found, in the second – 56.5 %.
Objective of the study was to identify the differences in the production of proteins in typical strain and genetically altered variant of V. cholerae O1, biovar El Tor.
Materials and methods. Natural strains M1062 (Astrakhan, 1970) and M1509 (Moscow, 2010) were used as model strains in this work. Strains were cultivated on Luria Bertani agar at 37 °C. Electrophoresis was performed in accordance with W.K. Laemmli technique (1970), mass-spectrometric profiling – the method described by A. Shevchenko et al.
Results and discussion. Mass-spectrometric scanning of cell lysates of the examined strains showed significant similarity of their proteomes (615 common proteins). The identified differences pertained to high expression of proteins in the strain M1062, participating in biosynthesis of DNA/RNA, included into “purines, pyrimidines, nucleosides” group, as well as regulatory proteins. In M1509 strain, biosynthesis of the proteins responsible for pathogenesis, adaptation under the influence of unfavorable environmental factors, included into “co-factors, vitamins, pigments” group, involved in lipid, carbohydrate, and protein metabolism, cellular processes, as well as proteins-transporters was increased. It has been suggested that the wide dissemination of El Tor genovariants is probably due to enhanced pathogenic and adaptive properties and also to the considerable transformation of cell metabolism.
Objective of the work was to investigate the peculiarities of gastrointestinal tract disorders in children with coronavirus disease caused by SARS-CoV-2 virus.
Materials and methods. We have analyzed the frequency, terms of emergence, duration and character of gastrointestinal disorders on the basis of medical records of 36 children receiving the treatment for community-acquired pneumonia of coronavirus etiology (positive for SARS-CoV-2 virus) in the children’s infectious disease department of Moscow State Clinical Hospital, Mytishchi. The majority of patients had moderate form of the disease (97 %), severe cases were not registered. The age range of the children – 8 months to17 years old. All patients were subjected to supplementary examination on Day 1 in accordance with the current recommendations. Children were treated in hospital settings for 8–15 days. Assessment of clinical status was performed at least once a day over the stated period, supplementary survey – at least once in a week, upon indications – more frequently. SARSCoV-2 virus was identified using polymerase chain reaction. All patients received standard therapy in compliance with advanced methodological recommendations, upon indications – other pharmaceutical products.
Results and discussion. Gastro-intestinal disorders in 23 children (64 %) emerged on Day 4–5, in 8 children (22 %) – before the start of the treatment; they lasted 3–5 days. In most of the patients diarrhea (64 %) and stomachaches (57 %) were observed. 67 % of the children demonstrated other symptoms indicative of gastrointestinal dysfunction. The disorders were characteristic for most of the children hospitalized with community-acquired pneumonia caused by SARS-CoV-2 virus. The symptoms are contained when the main course of therapy is conducted, indicated for treatment of COVID-19, and additional therapy recommended for management of gastro-intestinal infections is performed.
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