Put forward is a methodology for quantitative assessment of potential epidemic hazard (PEH) as regards mass events with international participation in terms of the relevant infectious diseases capable of (in compliance with IHR, 2005) inducing emergency situation of sanitary epidemiological character. It is established that PEH is an aggregate risk of infection importation by incoming contestants and visitors and of its dissemination across the mass even location. As a result of PEH level differentiation identified have been four following grades: high, medium, low and minimum. Calculated has been a rate of PEH for the XXVI Worldwide Summer Universiade in Kazan, 2013, exemplified by such infectious disease as cholera.

The paper discusses the opportunities of using universal model of local, evolving within a closed population, epidemics/outbreaks, developed at the premises of SRC VB “Vector”, for investigations and assessment of the impact of various resource limitations on the countermeasure effectiveness, and in particular isolation of a patient and contact tracing and isolation. Based on the epidemic dynamics analysis for a number of infections such as smallpox, anthrax, pneumonic and bubonic plague, hemorrhagic Ebola, Marburg, Lassa, Crimean-Congo fevers, it is demonstrated that occurrence of resource limitations of isolation can have a significant impact on the scale of epidemic aftermaths. Moreover, the outset of response measures takes a severe effect, as one of the important for localization of arthropod-borne infection factors is quarantine, alongside with isolation. The computer based software model is available at http://vector-epimod.ru

Analyzed is the management system for laboratory diagnostics of infectious diseases during the XXII Olympics and XI Paralympics, 2014 in Sochi. Reviewed is piece of work, executed in the pre-Olympic period as regards provision of laboratory facilities deployed for clinical material investigation, development of normative and regulatory documentation which considers peculiarities of the situation, identification of diagnostic capacities of the laboratories, and forecasting of the probable volume of laboratory studies by reference to various groups of infections. Put forward is the procedure for arrangement of work at the different stages of laboratory diagnostics, cooperation and response in case of emergency situation in the sphere of sanitary-epidemiological welfare of the population. Discussed is the role of geo-information system and current means of monitoring over epidemiological situation in the participating states and in the region of the Olympics in laboratory support organization.

Given is a general overview of the SAET performance during the XXII Olympic and XI Paralympic Winter Games, 2014 in Sochi. Discussed are the peculiarities of work management of the SAET diagnostic facilities; represented are the data on the structure and scope of laboratory investigations of clinical material and environmental samples. Analyzed is the experience of operation under major international mass event. Consequently, it is concluded that current SAET structure, its stuffing and equipping, the laid-up stock of preparations and test-systems have allowed for coping with a diverse task complex in the laboratory diagnostics of infectious diseases and indication of their agents.

Represented is the analysis and assessment of epidemiological and epizootiological situation on brucellosis in the Russian Federation in 2014. Human morbidity rates in 2014 exceed the indexes of 2013 by 7.7 %. Negative trend in the live-stock animal incidence (cattle and small ruminants), as well as in the numbers of registered index potentially hazardous territories, as regards brucellosis, remains. The highest brucellosis morbidity rates are in the North-Caucasian, Southern and Siberian Federal Districts. The quantity of brucellosis cases among the population has increased in Central and Far-Eastern Federal Districts. Put forward is the prognosis on the human brucellosis morbidity rates for 2015.

Based on the results of retrospective analysis of HFRS morbidity and epizootiological data on the numbers and contamination of mouse-like rodents, revealed has been epidemic potential shift of the HFRS foci. It requires reconsideration of the scope, format, and objectives of preventive and anti-epidemic activities. Periodicity of fluctuations in the numbers of small mammals, habitant in the territory of the Ulyanovsk Region, makes up three years on average. Since 2011 up to 2013 generalized depression in the abundance rates was observed. The period was marked by the reduction of the segment of main Hantavirus carrier - common red-backed vole. It was the key factor for epidemic process activity decrease, which in its turn resulted in the decrease of morbidity rates among the population. Therewith, the transformations that affect the population of mouse-like rodents are the determinative factors in the evolution of epidemic process which shapes epidemiological situation on HFRS in the territory of the constituent entity.

Natural tularemia foci in the territory of the Khanty-Mansiisk Autonomous District - Yugra are still active and sustainable ones despite of the extended periods of epidemiological welfare. Agent reservoir is the water vole. However, recently tularemia agent has been detected in the northern red-backed vole and common red-toothed shrew. Khanty-Mansiisk town, Berezovsky, Khanty-Mansiisk, Kondinsky, and Oktyabr’sky Regions are situated in the territory of the most active foci. Serious outbreaks of infection among the population occur with an interval up to 30 years. Their transmissibility preconditions the scale and intensiveness. All cohorts are under the risk of exposure, irrespective of the age and occupation, including the infants. The paper discusses tularemia outbreak impact on the population of Khanty-Mansiisk and the Khanty-Mansiisk Region in July-October, 2013. Casualty toll is 1005 cases. Given is a brief retrospective analysis of epidemiological situation on tularemia since 1930-s up to the onset of the outbreak under investigation. Described is the experience in management of the response activities.

The territory of the Crimean Peninsula is an endemic one as regards various natural-focal infections. The paper contains the data on epizootiological survey of 8 administrative regions of the Crimean Federal District (CFD). Carried out has been small mammals and Ixodidae ticks census. Identified is their species composition. Performed have been the field samples collection and laboratory studies for the presence of bacterial and viral infection agents. Established is the fact that H. marginatum is a dominating species among the ticks, collected during the spring season, while the house mouse prevails among the small mammals. As for the autumn collection of samples, the dominating species are H. punctata and common vole, respectively. Laboratory investigation results indicate the circulation of the agents of Ku fever, tick-borne spotty fever, Ixodidae tick-borne borreliosis, human granulocytic anaplasmosis, monocytic ehrlichiosis, hemorrhagic fever with renal syndrome and leptospirosis.

In consequence of epizootiological investigation, differentiated has been Kazan territory according to the potential epidemic hazard levels; determined is the abundance of carriers and vectors of natural-focal infection agents in and around the territory of the Summer Universiade-2013; substantiated is the scope of preventive measures (disinfection, deratization, desinsection), as well as their frequency and coverage area. Timely detection of the regions with expressed epizootic potential growth has made it possible to carry out prophylactic activities and reach an epidemic-hazard-lowering effect as regards particular territories of the natural and anthropourgic infectious disease foci. Further on, validated has been disinfection as part of specific prophylaxis and as a basic measure for elimination of risks of the population exposure to natural-focal infections directly during the mass event.

Demonstrated is the structural and functional interrelation between sanitary protection of the territories, epidemiological monitoring, and the state sanitary-epidemiological surveillance. Sanitary protection of the territories (SPT) of the Russian Federation is a separate focus area authorizing sanitary prophylactic (anti-epidemic) activities, with a self-sustained sub-goal. It combines individual roles appertained to epidemiological monitoring and to federal sanitary-epidemiological surveillance, only in reference to a particular item - emergency situation in the sphere of sanitary-epidemiological welfare of the population. Specificity of the SPT is a balance between functionality and capacity of the monitoring, surveillance and control. Legislative and regulatory legal acts, which concern sanitary protection of the territory, provide the means for defining its basic organizational principles: country-wide activities, self-sufficiency and self-support of the Rospotrebnadzor, three-level hierarchical system, and information exchange.

Carried out has been comparative analysis of the cell antigen immune-electrophoregrammes of B. pseudomallei C-141, B. mallei 10230, B. thailandensis 264, and B. cepacia 25416 after immunoblotting with immune rabbit sera to living cells of avirulent B. pseudomallei 107, celllular and extracellular antigens of B. pseudomallei 107, B. pseudomallei C-141, B. mallei 10230, B. thailandensis 264, and B. cepacia 25416. Visual investigation has revealed the presence of fractions with molar mass of 18.4 and 35 kDa in B. malei 10230 cell antigens’ electrophoregramme obtained by means of immunoblotting with serum to B. malei 10230 cell antigens, as distinct from B. pseudomallei C-141 electrophoregramme with the same serum. This makes it possible to distinguish the pathogenic Burkholderia. At the same time utilization of heterologous immune sera enhances discriminating capacity of the comparative assay. The level of reproducibility of immune-electrophoregrammes allows for the deployment of the similarity coefficient for computer-based comparative analysis.

Provided are the data on the management and carrying out of the laboratory control over the hot-water supply systems of the sports venues and accommodation areas for the presence of Legionella pneumophila during the XXII Olympics and XI Paralympics in Sochi, 2014. Inspected have been 105 facilities. The samples from 37 of them showed positive. L. pneumophila DNA concentration varies between 2.19·10² and 3.92·10⁷ genome equivalents (g.e.)/l. By means of bacteriological investigation detected have been legionella colonies in the water supply systems of 16 facilities, at one of the items - the loading is over 1·10⁴ colony-forming units/l. Performed has been comparative analysis of the results obtained using PCR assay and bacteriological test. Based on the genotyping of 7 isolated strains serogroup 1 and 2 strains of 2-14 serogroups it is concluded that L. pneumophila strains circulating in the resort town Sochi are genetically heterogeneous.

Developed is the method for tularemia microbe DNA detection using PCR with electrophoretic and hybridization-fluorescent registration of results. iglBC genes have been chosen as DNA-matrixes, being species-specific ones for tularemia agent. Based on the results obtained constructed have been preparations for tularemia microbe DNA detection in biological material and environmental samples applying PCR with electrophoretic registration of results and real-time PCR: “Gene Francisella tularensis - REP” and “Gene Francisella tularensis RHF”, respectively. Identified are the package contents to be included into the test-systems. Sensitivity and specificity of the designed panels are validated through investigations of tularemia agent bacterial emulsions and suspensions from small mammals’ organs, from ticks, fleas and mosquitoes, as well as through studies of soil and surface water samples, sputum and human blood probes, experimentally contaminated with tularemia agent. Test-systems demonstrate high sensitivity (1·10³ microbe cells/ml) and specificity (100 %), irrespective of the type of test material.

The paper contains the data on isolation of Francisella tularensis cultures from humans and rodents during tularemia epidemic in Khanty-Mansiisk in August-September, 2013. All the obtained F. tularensis strains (six cultures of tularemia microbe from patients, and four cultures - from small rodents caught in the territory of the city and its suburbs) fall under Holarctic subspecies, are highly virulent for the laboratory mice, resistant to erythromycin, ampicillin, and cephalosporin, but sensitive to amikacin, tetracycline, doxycycline, and gentamycin. Using 25-locus-MLVA typing it is demonstrated that the strains under study form a common cluster on the phylogenetic tree along with F. tularensis strains previously isolated in the territory of the Southern Urals and Kazakhstan. All the strains from the foci are closely related ones, and based on the investigations of hyper-variable Ft-M3 locus, fall into four groups. The strains, isolated from humans, combine into three groups, and the strains isolated from rodents - into two, and notably that one of the groups of Khanty-Mansiisk cluster comprises the strains isolated from both humans and rodents.

Carried out has been whole-genome sequencing of the Vibrio cholerae 81 El Tor Inaba strain DNA, isolated from Temernik River waters in the territory of Rostov-on-Don in July, 2014. Identified is the presence of hybrid CTX prophage, which contains ctxB gene of the classical type ( ctxB1 allele), as well as rstR gene El Tor type, tcpA gene with the mutations in coding and promoter regions ( tcpET^CIRS allele), the null-mutation in rtxA gene, and extended deletion inside the pandemicity island VSP-II. All the stated above peculiarities are characteristic of cholera vibrio genovariants, which have an enhanced epidemic potential and have recently replaced the typical strains of this biovar.

Coronaviruses are enveloped viruses with a single-strand “+” RNA, its genome size varying from 25 to 32 thousands of nucleotides. They cause respiratory and intestinal diseases in animals and humans. The review contains the data on human infection cases induced by a new coronavirus (NCoV), as well as the information about probable natural agent reservoirs, mechanisms of transmission, some characteristic features of the etiological agent, methods of diagnostics and identification, complete genome sequence, and NCoV relation to the established coronaviruses.

Studied has been the state of the nucleolar apparatus in lymphocytes of periphery lymphoid organs in laboratory animals vaccinated against plague and tularemia in order to evaluate predictive capability of the parameter in the context of pre-clinical evaluation of the vaccines under development, against particularly dangerous infections. For the detection of silver adjoining proteins, contained in the nucleolar organizer region (AgNOR), W.M.Howell and D.A.Black staining of semi-thin paraffin sections of lymphoid organs of the immunized BALB/c mice has been performed. For this purpose, ready-assembled panel, manufactured at BioVirtum, has been deployed. Under assessment of the functional state of immunogenesis organs in vaccinated against plague and tularemia animals revealed has been the increase in numbers of cells with three and more AgNOR-positive nucleoli in the nucleus of the lymphocytes of periphery organs of the biomodels’ immune system. It indirectly manifests proliferative capacity of immune system cellular component. Informativity of the quantitative evaluation of the cell proliferative activity by the reference to the status changes of AgNOR in the nuclei of immunogenesis organs’ lymphocytes makes it possible to entertain the possibility of introducing this method into the existing morphological vaccine quality assessment scheme at the stage of the pre-clinical trials.

Put forward is an efficient method for manufacturing cholera toxin B-subunit. Its advantages are relative simplicity and economy feasibility, as well as maximum output of the purified B-subunit, absolutely free from toxic A-subunit contaminant. All this is due to the deployment of cholera vibrio recombinant strain producing only cholera toxin B-subunit instead of cholera toxin as it is, which results in lack of residual preparation toxicity. Applied has been gel-penetration column chromatography, providing for stable native state and maximum antigen output. The method under discussion is verified experimentally. Sample purity has been analyzed after each phase of chromatographic investigation on TSK gel HW-60, using disc electrophoresis. It is established that three steps of purification are ample for the obtainment of cholera toxin B-subunit preparation free from admixtures. Immunological activity of the purified B-subunit is validated by monoclonal antibody obtainment. Designed preparation of cholera toxin B-subunit and monoclonal antibodies to it can serve as a basis for the development of various immune-diagnostic test-systems alternatives.

Carried out is comparative analysis of adsorption activity of the constructed specific enterosorbent as regards pharmacological markers - methylene blue, bovine serum albumin, gelatin, and neutralizing activity in vivo with reference to the specific marker - cholera toxin. The data obtained testify to the fact that the level of the sorption activity in relation to the utilized marker substances both of initial micro-particles of chitosan, and its specific modification, is comparable to the corresponding parameter of the reference drugs. It is established that the process of immunoglobulin component immobilization initiates the changes in electro-chemical properties of the chitosan active surface, leading, on the one side, to the decrease in the sorption activity of the specific preparation in relation to methylene blue and gelatin, and on the other, to the increase in the protein marker sorption activity - bovine serum albumin, as compared to the initial chitosan matrix. In the course of the toxin-neutralizing capacity investigation, identified has been high anti-toxic activity of the experimental preparation in reference to cholera vibrio enterotoxin. The results attained give grounds to view the constructed anti-enterotoxic enterosorbent as potential prototype of efficient aids for cholera prophylaxis and treatment.

Investigated have been 8 cultures of Francisella tularensis strain 15 NIIEG (lyophilized in 1953, 1966, 1969, 1987, 1990, 2003, 2012, and 2013, respectively) stored at the State Collection of Microorganisms of the Scientific Center on Expertise of Medical Application Products. It is established that the majority of cultures has maintained their immunobiological properties. However, it is of note that liophilization does not prevent F. tularensis strain 15 NIIEG from changes in its residual virulence under extended storage. Revealed is the fact that LD₅₀ for 7 cultures of tularemia microbe strain is within the limits of 100-250 microbial cells (m.c.). At the same time, residual virulence for the strain which dates back 1966 is 7.3·10⁵ m.c. Immunogenic activity rates in F. tularensis 15 NIIEG strain cultures range within specified limits. Apart from this, F. tularensis 1987 strain does not comply with the established requirements to the “specific safety”, as subcutaneous inoculation with 5·10⁹ m.c./ml caused death of Guinea pigs within the scheduled observation time. Demonstrated is the necessity in maintaining constant stability of the original immunobiological properties in Francisella tularensis strain 15 NIIEG under extended storage conditions.

Carried out have been two experimental studies on hybridization of mouse myeloma cells and lymphocytes of BALB/c mice immunized with inactivated microbe Francisella tularensis cultures. As a result obtained have been hybridomas-producers of monoclonal antibodies (MAb) specific to the antigens of tularemia agent. Evaluated have been the prospects of its application for the detection of the agent under discussion using enzyme-linked immunoassay. Established is the fact that monoclonal antibodies produced by 31G1F10, 32E5D3, 35B11C8, 36C2F11 hybridomas make it possible to identify microbe cells of various tularemia agent strains when concentrated up to 0.5·10⁶ mc/sm³, and do not interact with cultures of heterologous microorganisms when concentrated to 1.0·10⁸ mc/sm³, which testifies to their specificity. These MAb are planned to be used for the construction of immune-enzyme and immune-chromatographic test-systems designed for tularemia agent detection.