The paper describes the International Health Regulations (2005), hereinafter referred as IHR (2005), by the example of SARS, A/H5N1/, A/H1N1/09, Zika fever, and MERS-CoV control as an effective tool for prevention and control of public health emergences (ES of sanitary-epidemiological character) of international concern. Late detection of Ebola fever epidemic in West Africa (2013–2016) and announcement of public health emergency of international concern at the time when epidemic up-scaled to a threat to national security of the affected countries and high-priority threat to global community (ES of biological character) is attributed to non-use of methodological capacities of IHR (2005) on verification of ES at an early stage of epidemic development because of unpreparedness of local, regional, national and international public health services to such an epidemic situation. WHO plans to achieve scientifically-based reduction of the time from the onset of epidemic events to the effective response measures. Russian Federation, scientifically implementing IHR (2005) from the very beginning and having devised methodology of IHR implementation on the national level, has a strong premise for scientific provision of this process. 

Objective of the study is to comprehensively evaluate and forecast the development of epidemic process in case of anthrax in Altai Territory. Materials and methods. The data on stationary hazardous as regards anthrax areas (SHA) were obtained while analyzing cadastres of SHA in Altai Territory, as well as the reference book “Cadastre of stationary hazardous as regards anthrax areas in the Russian Federation”. Statistical and report forms from the institutions of the Rospotrebnadzor and Federal Service for Veterinary and Phytosanitary Surveillance in Altai Territory, and from Altai Plague Control Station were used for retrospective analysis of anthrax morbidity rates in Altai Territory over the period of 1953–2015. Laboratory investigations of field and clinical samples were carried out in compliance with Methodological Regulations 4.2.2413-08. Results and conclusions. Anthrax remains topical issue for Altai Territory as there are 1262 stationary hazardous areas situated in here, as well as registered animal and human infections. The density of such areas in Altai Territory is 7.5 times higher than across Siberia. Factors for the SHA formation and their sustainability are soilclimatic aspects of the region. The majority of hazardous areas are found in steppe and forest-steppe zones, predominantly along the cattle-driving paths. In Altai Territory, between 1953 and 2015, out of 1262 registered areas, 314 ones manifested epidemiological activity, including 246 new ones. Human infections were reported in 36 districts (59.1 %), while in 21 (34.4 %) – morbidity rates exceeded the territorial average. Cumulative dynamics of anthrax morbidity as regards livestock in Altai Territory in 1953–2015 showed downward trends. Complex evaluation of the situation and forecast of epidemic process development, employing composite index – epizootiological index and zoning of the territory according to the risk of epidemic complications – allows for the improvement of managerial decisions with the view to adequate anti-epidemic measures. 

123 cases of human infection with Francisella tularensis were registered in the territory of the Russian Federation in 2016. 90 % of the total number was reported by three Federal Districts: North-Western, Siberian, and Central ones. Epizootic manifestations of varying intensity were detected in 56 regions. A considerable number of tularemia cases were observed in the Republic of Karelia, Nenets Autonomous District, Omsk and Ryazan Regions, and in Saint-Petersburg. Conspicuous are some recent peculiarities of tularemia manifestations: non-specificity of clinical symptoms, especially at early stages of the disease, their increasing variability against the background of mixed infections with tularemia and other natural-focal infections agents. The most expressed symptoms are fever and lymphatic adenitis; the patients are diagnosed with tularemia during initial days of the sought treatment in 40-45 % of the cases. Based on the data on morbidity rates in humans, epizootiological condition of natural tularemia foci, the numbers of isolated cultures and incidence of the vectors and carriers of the agent, as well as taking into consideration the scope of preventive vaccination in the regions, identified were the territories in which epidemic complication are most likely to occur in 2017. 

Objective of the study is to locate the present-day sites and sizes of the natural plague foci in the Volga-Ural Interfluve. Materials and methods. According to the results of the field and office mapping, 2015 in the Volga-Ural steppe and sandy plague foci, designed have been electronic maps of the sectors situated in the periphery. Results and conclusions. During the past 3–4 decades significant decrease in plague enzooties in the territory of the Volga-Ural Interfluve has been observed. It occurs due to the plowing and melioration of the grazing lands. It is established that the southern and south-western outskirts of enzootic territories have natural boundaries represented with the linear elements of hydrography, and the eastern ones are the state border of the Russian Federation and the Republic of Kazakhstan. At sites where suitable linear elements of the landscape are absent (up north and west) the frames of the sectors, where evidence of enzooty is retained, are used as the formal external boundaries. Widespread plowing of semi-desert area made the territories in the eastern part of the Volgograd Region inadequate for small souslik habitation, which marks the loss of plague enzooty features. This fact in its turn led to the reduction of steppe focus acreage by 30 %, the coverage being currently 20873 km2 . Configuration of the sandy focus changed little, if at all, and amounts to 8625 km2 . Registering of the actual position and sizes of natural plague foci within the strictly set boundaries put on topographic maps provides for targeted planning and complex prophylactic anti-plague measures. New spatial indicators of natural plague foci are formalized in official normative-methodological documents regulating epidemiological surveillance over plague. 

The article presents analysis of human and animal incidence of brucellosis in the Russian Federation in 2016. Epidemiological situation on brucellosis is characterized as unstable. Outlined has been the decrease in the number of newly diagnosed brucellosis cases in humans (15.8 %) against the background of the increasing trend for distress and deterioration of the epizootiological situation on brucellosis among the cattle in the regions with well-developed animal husbandry. Cases of group infection among the population in three constituent entities (Stavropol Region, Samara and Leningrad Regions) have been registered, which are of sporadic nature. According to the Federal Service for Veterinary and Phytosanitary Surveillance, mid-term forecast of epizootiological situation on brucellosis in Russia is unfavorable, which under the current trend constitutes a real risk of widespread brucellosis among the livestock in the regions of the Central and Volga Federal Districts. In 2017, we can predict maintaining of the unstable situation on the disease, morbidity rates among the population varying within the range of 340–370 cases. 

Objective of the study is to conduct the complex investigations aimed at identification of circulation pathways of tularemia agent, as well as to define the main carriers and vectors of the disease. Materials and methods. Trap trench method, trap fences and trap-lines were used to catch small mammals for examination. Bacteriological testing on tularemia infection in animals was carried out using spleen samples. In addition, water samples were studied. Results and conclusions. In 2015, performed were complex zoological-parasitological and bacteriological studies aimed at identification of circulation pathways of tularemia agent in the natural focus of floodplain-river type in the Khanty-Mansiysk Autonomous District – Yugra, in the confluence of Irtysh and Ob rivers. The total volume of the material tested was 542 specimens of small mammals of 16 species and 447 specimens of amphibians belonging to 4 species. It was established that, compared to 2013, zoocoenosis restructuring in the floodplain habitats occurred. The main carrier and a massive source of tularemia infection – the water vole – was excluded from the small mammals’ community. Also, ectoparasites composition changed; no specific ectoparasites of the water vole were to be found. In 2015, in the confluence of Irtysh and Ob rivers, lukewarm epizooty among the small mammals was observed. Essential for the occurrence of acute tularemia epizooty prerequisites were absent. 

Objective of the study is to design the DNA-microarray for differentiation of Y. pestis strains of the main and non-main subspecies and biovars of the main subspecies. Materials and methods. Efficiency analysis for the devised means was conducted using 62 Y. pestis strains of various subspecies and biovars, isolated in the natural foci of Russia and neighboring countries. Results and conclusions. Selected have been the DNA-targets, probes and primers – calculated. Enhanced is the method of sub-specific and biovar differentiation of Y. pestis strains by means of DNA-microarray. DNA-chip with “Med24”, “glpD(-93)”, and “45” targets allows for prompt differentiation of the strains of the main and non-main subspecies and biovars of the main subspecies based on the presence and absence of fluorescent signal by the specific for the main subspecies and its biovars DNA-targets. 

Currently, bacterial small RNAs with regulatory functions are characterized as a heterogeneous claster of highly structured single-stranded RNAs which usually would not be translated into proteins. A large group of small RNAs consists of small regulatory RNAs functioning through the mechanism of complementary interaction between small regulatory RNA bases and bacterial messenger RNAs (mRNA). The review provides the description of direct participation of small RNA mechanisms in realization of pathogenic properties, biofilm formation and virulence regulation in V. cholerae strains. In particular, the Hfq-dependent small RNAs that control the expression of genes responsible for virulence and biofilm formation of V. choleraе, and the impact of small RNA on the secretion system of VI type are presented. We also characterized the small RNAs–controlled process of V. cholerae vesicles formation, which have a great significance for colonization ability of cholera vibrios. In addition, such method of regulation at RNA level as “riboswitches”, (RNA switches) is described. 

Objective of the study is to apply 2D-electrophoresis for imaging protein spectrum of exoprotein fractions, as well as identification and comparison of biomarker expression of the key pathogenicity factors in plague and cholera agents. Materials and methods. 2D gels of exoprotein fractions were obtained on the model of Vibrio cholerae Inaba 569B, V. cholerae El Tor M888Ctx+ and Ctx– strains, and also Y. pestis EV NIIEG strain. Capsular antigen F1 and cholera toxin were used as biomarkers of major exoproteins of plague and cholera agents. Results and conclusions. While studying cholera toxin through 2D-electrophoresis on IPG strips with pH 3–10 gradient, the antigen fell into a number of protein spots, two of which were close to A1 domain parameters (MW 20.309 kDa and pI 6.51) and B-subunit monomer (MW 11.091 kDa and pI 7.68). Comparative protein spectrum analysis of the exoprotein fractions of V. cholerae Inaba 569B, V. cholerae El Tor M888 Ctx+ and Ctx– has revealed the spots with similar parameters, excluding the strain V. cholerae M888 Ctx– . Investigation of 2D gels of capsular antigen and exoprotein fractions of Y. pestis EV strain has demonstrated that protein spots corresponding to the parameters of the subunit form and diameter of the antigen are present in 37 °C culture patterns. 28 °C culture sample contains F1 subunit in a far lesser concentrations. On the model of toxigenic V. cholerae Inaba 569B strain, isogenic system of V. cholerae El Tor M888 Ctx+ and Ctx– strains, and Y. pestis EV NIIEG strain, high efficiency of 2D-electrophoresis in protein spectrum construction for exoprotein fractions of the cultures, as well as identification and comparison of biomarker expression of major exoproteins of plague and cholera agents is established. 

Biological method of investigation is specified for the laboratory diagnostics of plague. Mastering of this method by the trainees within the frames of further vocational education is associated with the use of avirulent Yersinia pestis strains and vaccine Y. pestis strain EV line, which while providing safety does not allow for typical pathomorphological pattern on biomodels, as well as for isolation of microorganisms from internal organs. Objective of the study is to select avirulent Yersinia pestis strains and to conduct comparative analysis of the simulation techniques for plague on biomodels. Materials and methods. Utilized were Y. pestis strains. Virulence was evaluated both, in vitro (polymerase chain reaction) and in vivo (LD50 for white mice). Results and conclusions. Set forward have been avirulent Y. pestis strains, prospective in terms of mastering biological method of laboratory diagnostics of plague, and means of their application for simulating plague in biomodels. The designed approach allows for exercising biological methods of plague investigation to the fullest extent, enhancing biological safety of practical studies and reducing the time line for isolation and accumulation of pure bacterial culture. 

The Arenaviridae family consists of a large group of single strand ambisense RNA viruses that are separated phylogenetically, serologically and geographically into Old World and New World viruses. Recent studies indicate that cellular entry of arenaviruses requires a series of cellular protein interaction and molecular mechanisms. The arenaviruses entry into cells is initiated by the interaction of viral glycoprotein with one or more receptors on the surface of host cells. The main host cell factors that are involved in filovirus entry are attachment factors (α-dystroglycan for Old World and human transferrin receptor 1 for New World viruses), endolisosomal host cell factors (cathepsins B and L and Niemann-Pick C1 protein). The review presents the modern knowledge about the role of structural proteins of arenaviruses and some cell factors in pathogenesis of the diseases, caused by arenaviruses. 

Objective of the study is to evaluate the influence of CTXφ prophage deletion, which carries ctxAB genes, on phenotypical properties associated with pathogenicity or biofilm formation in non-toxigenic mutants. Materials and methods. Utilized have been the clinical strains of Vibrio cholerae biovar El Tor and their spontaneous non-toxigenic mutants that lost CTXφ prophage. Applied have been microbiological and biochemical methods, inoculation of model animals with cells of the strains under study. Results and conclusions. The results of comparative analysis of phenotypic properties in isogenic toxigenic and non-toxigenic strains of Vibrio cholerae biovar El Tor, which lost CTXφ prophage encoding the cholera toxin, are represented. It is established that the deletion of CTXφ prophage leads to the simultaneous change of several phenotypic properties associated with virulence (colonizing ability, production of soluble hemagglutinin/protease and heat labile hemolysin/cytolysin) and biofilm formation (motility, exopolysaccharide biosynthesis) in spontaneous non-toxigenic mutants. It is suggested that the reason for these phenotypic changes in the mutants might be the changes in activity of the related to each other regulatory genes controlling virulence and biofilm formation process in cholera agent.

Objective of the study is to specify the character and dynamics of morpho-functional changes in the organs of peripheral immune system of BALB/c mice in case of immunization with vaccine Yersinia pestis EV NIIEG strain against the background of immune modulation. Materials and methods. Applying a combination of histological, histo-chemical, immune histo-chemical methods, and morphometric analysis, defined has been the targeting of immunological processes in peripheral lymphoid organs of BALB/c mice. Results and conclusions. Revealed has been stimulating effect of polyoxidonium in concomitant use of Yersinia pestis EV NIIEG, on the processes of cell proliferation in T-zones of lymphoid organs, on activation of T- and B-lymphocyte sub-population in them by reference to the increment in the amount of specifically stained with CD25 marker cells and nuclei of lymphocytes with high argyrophil granule content, indirectly characterizing the functional state of nuclei apparatus, as well as on response intensification of neuroendocrine environment of lymphoid structures. Based on the results of complex morphological analysis, specified have been the peculiarities of morphofunctional changes in immune competent organs of biomodels, being indicative of polyoxidonium potential to increase the effectiveness of vaccination against plague. 

F. tularensis ssp. novicida, considered earlier as a representative of a separate species, has been recently classed among F. tularensis variety, based on the results of comparative analysis of 16S-ribosomal RNA. Subspecies novicida can cause disease only in immunocompromised humans and is low virulent for rabbits. Despite this, high rate of homology of the nucleotide sequence of F. tularensis intraspecific taxon is established. Objective of the study is to obtain protective surface antigen complex from F. tularensis ssp. novicida Utah 112 (ATTC 15 482) cells and investigate its properties. Materials and methods. Protein, carbohydrate, and lipid content of the antigen preparation was measured using conventional colorimetric methods, SDS-PAGE was conducted according to U.Laemmli, and immunoblotting – to H.Towbin. For purification and molecular mass determination column chromatography was applied. Immune-chromatographic activity was analyzed by immune-enzyme assay. Immunogenicity of the produced preparation was tested on scrub white mice, with LD50 and ED50 calculated according to Karber’s method. Results and conclusions. Carried out has been comparative analysis of physical-chemical, antigenic and bio-chemical peculiarities of the protective antigen complex obtained from F. tularensis Utah 112 cells and equivalent antigen complex obtained from the vaccine strain – F. tularensis 15 NIIEG. Protectivity of the preparation has been tested through inoculation of the immunized white mice with virulent F. tularensis 503/840 strain. Demonstrated have been distinctive features of the new preparation, by structure and composition, as compared to similar antigen from the vaccine producer strain, as well as the slowdown of its immunochemical and protective activities. 

Objective of study is to prepare hybridomas-producers of monoclonal antibodies to brucellosis agent antigens. Materials and methods. B. abortus, B. melitensis, B. suis strains from the State collection of microorganisms of the 48th Central Research Institute Affiliated Branch and BALB/c mice. Hybridization was performed as described by G.Kohler and C.Milstein in modification by Fazekas De St. and Scheidegger D. The study of specific activity of immune sera, hybridoma supernatants, ascites fluid, and monoclonal antibody preparations was performed using ELISA. Results and conclusions. Obtained and characterized have been hybridomas-producers of monoclonal antibodies to specific antigens of brucellosis agent. They are active and stable antibody producers in the repeated passaging both, in vitro and in vivo. Obtained have also been the ascites fluid and preparations of monoclonal antibodies of brucellosis agent. Carried out has been substantiated selection of antibodies which could provide for the most sensitive ELISA. It is established that the monoclonal antibodies produced by hybridomas 232B6H7, 232G12F7, 233B2C5 in combination with brucellosis rabbit immunoglobulins allow for the identification of microbial cells of type strains of various Brucella species in concentrations ranging from 0,25·10⁶ mc·sm^–3 up to 1,0·10⁶ mc·sm^–3 and gave negative results with cultures of heterologous microorganisms in the contents of 1,0·10⁸ mc·sm^–3. Hybridomas-producers of monoclonal antibodies are planned to be used for the construction and manufacturing of immunodetection test-systems.

Smallpox eradication due to global vaccination is still one of the paramount triumphs of medical science. Given the termination of the subsequent immunization, nowadays humanity virtually possesses no antivariolic immunity and is unprotected against the pathogenic for humans orthopoxviruses. Utilization of the first-generation traditional live vaccines, obtained with the help of the virus replication on calf skin, or the second-generation preparation, produced in mammalian cell cultures or grown on bird embryos, for mass vaccination is currently unacceptable in view of considerable increase in immune deficiency states among the human population within the recent decades. Attenuated non-replicating antivariolic vaccines of the third generation, obtained in the process of multiple vaccinia virus (VV) passage on cell cultures of heterologous host, induce weaker antivariolic response as compared to traditional vaccine. The most prospective approach is to produce the vaccines of the fourth generation, applying targeted VV genes’ mutation, which control protective reactions of an organism against viral infection, as well as host range genes and the genes involved in nucleic acid metabolism, while skipping the genes responsible for virus replication. Novel attenuated and highly immunogenic strain, VV LIVPΔ6, having mutations in 6 virulence genes, is presently in the phase of pre-clinical trial and later on it may turn an effective and safe vaccine of the fourth generation against smallpox and other orthopoxvirus infections in humans. 

Objective of the study is to prepare rabbit monospecific anti-abortus serum to a Brucella S-form thermoextract, as well as to evaluate its activity and specificity. Materials and methods. Utilized was polyvalent hyperimmune serum obtained to Brucella thermoextract. It was adsorbed by a strain of heterologous species Brucella melitensis 16 N. Reference, vaccinal and field Brucella strains were used to estimate the activity and specificity of the serum produced. Results and conclusions. This serum possesses high activity (1:160, 1:320 titer) and specificity, and the results completely coincide with the data specified in the table of differential properties presented in МR 3.1.7.1189-03 “Prophylaxis and laboratory diagnostics of human brucellosis». The outlined method for manufacturing monospecific Brucella anti-abortus serum permits for a considerable decrease in biological hazard for personnel involved in the technological operations. 

The paper describes an effective liquid nutrient medium, utilization of which in the process of submerged cultivation of the vaccine tularemia microbe strain allows for the production of high concentrations of viable biomass with low rates of dissociation, which is essential in manufacturing of live vaccines. Dry enzymatic hydrolysate of fibrin, by-product of anti-rabies immunoglobulin production is used as a nutrient-rich base of the new nutrient medium. 

Objective of the work was to determine protective potential and safety for animals of the naturally attenuated B. anthracis 363/11 monoplazmid non-capsular strain to be used as a new candidate vaccine strain for the development and enhancement of specific prophylaxis of anthrax. Materials and methods. Vaccine strain 55-VNIIVViM that is used for welfare provision as regards anthrax in animals in the territory of Russia was employed for the comparison of protective properties. Identification of LD50, ImD50 and protective efficacy was carried out using standard protocols for modeling the infection on laboratory (white mice, guinea pigs) and targeted (sheep) animals. Results and conclusions. It is established that avirulent for white mice B. anthracis 363/11 strain demonstrates higher protective and immunogenic activity than 55-VNIIVViM strain. It does not reverse to the virulent variant after passaging on the nutrient media and in the organism of susceptible animals. Thus it allows for considering B. anthracis 363/11 strain as a promising candidate vaccine strain, the utilization of which will improve epizootic situation on anthrax in the Russian Federation. 

To the 70-th Anniversary of Zurab L. Devdariani.

Of blessed memory of Lyubov V. Samoilova.