Considered are main hypotheses that explain plague natural focality. The modern advances in the investigation of plague microbe genetic structure and its biofilms are demonstrated to play determinative role in interpretation of plague enzooty mechanism. Possible role of the nematodes in the transfer of plague microbe biofilms to the flea larvae is discussed. Considered is epizootiological significance of plague microbe trans-larval transfer for implementation of its vertical transmission from the soil biotope to the organism of warm-blooded animals. Analysis of flea larvae obtained in locations where plague persists, for the presence of plague agent, seems to be a promising approach as it can detect the readiness of the parasitic system of the natural focus for emergence and development of the epizooty.

Carried out is the analysis of scientific-methodological approaches to the development of algorithm for epizootic and epidemiological investigation of anthrax cases in the administrative territory. Put forward are five criteria of analysis for epidemiological investigation of anthrax sporadic cases and outbreaks, in order to develop the package of administrative, informational, anti-epizootic, anti-epidemic, and preventive measures, aimed at localization and elimination of anthrax focus.

Presented are the results of ecological and epizootiological surveillance of the territory of the Saratov region, which was carried out in autumn of 2010. The surveillance was aimed at detection of West Nile (WN) virus circulation and premises for WN Fever natural focus formation. It is demonstrated that in 2010 WN virus circulation took place in damp biotopes of the Saratov region territory, and that common species of small mammals were involved in it. Presented are the results of analysis of the WN virus role in the infectious pathology in the territory of the Saratov region.

The analysis of current regulatory requirements and applicable guidelines, as well as scientific papers, confirmed the existence of the subsystem of the control over epidemiological situation at the potentially hazardous biological facility. This subsystem is an element of the facility biosafety support system. Developed is the set of models which contain schemes of epidemiological situation management in case a sick facility officer is detected whose disease is suspected to be caused by the microorganisms of I-II pathogenicity groups.

Considered is the experience of development of the decision support system (DSS) in the sphere of biological safety provision. Described are the objectives, functions, and architecture of DSS. Represented are the results of operational program-testing in the model territory (the Astrakhan region). Indicated is the effectiveness of DSS for information support of the control activity over internal and external biosafety hazards. Determined are the directions for further development of DSS.

Studied are diagnostic properties of four anthrax bacteriophages. Indicated are various spectra of specific lytic activity and specificity of bacteriophages Gamma A-26, K VIEV, VA-9 and Fah-VNIIVV&M. K VIEV, VA-9 bacteriophages with wide spectrum of specific activity possess relatively low specificity. On the contrary, Fah-VNIIVV&M bacteriophage with high specificity exhibited narrow spectrum of specific lytic activity. Gamma A-26 bacteriophage is proposed for application as a diagnostic one, as it lyzes B. anthracis strains of all types, its specificity being equal to 90 %.

Identified are the occurrence, serotypic and genotypic variations of Hepatitis B virus isolates (HBV) among the Novosibirsk region inhabitants (n=2000), native population of the Alarsk District of the Irkutsk Region (n=487) and Shuryshkarsk Township of the Yamalo-Nenets Autonomous District (n=657). Occurrence rate of hepatitis В surface antigen (HBsAg) among different groups of the Novosibirsk Region population varied within the limits of 3,6-35,0 %. It was 8,2 % in Alarsk District, and 3,2 % in Shuryshkarsk Township. HBV isolates of D genotype (92-97 %) prevail among the population of Siberia; few are the cases of A (1,7 %) and C (1,2-8 %) genotypes. The identified varying occurrence of HBV sub-genotypes and HBsAg subtypes in two aboriginal groups of Siberia (D3 sub-genotype and ayw2 subtype - in the Alarsk District, D2 and ayw3 - in Shuryshkarsk Township) suggests the existence of, at least, two isolated HBV virus populations, circulating among different groups of Siberia native population .

Studied is the UV irradiation and nalidicsic acid effect on the RecA-protein synthesis in Francisella tularensis 15/10 cells. Obtained is the specific murine serum to the recombinant RecA-protein. The results of immunoblotting with this specific serum demonstrate that the amount of RecA-protein in Francisella tularensis 15/10 cells is not increased after the exposure to these damaging factors.

The studied strains of avian influenza virus A/H5N1 (AIV), isolated in the territory of the Russian Federation and CIS countries, can be classified into three groups according to their virulence for mice after intranasal and aerosol challenging: highly virulent strains [50 % aerosol lethal dose (ALD₅₀) - <3,0 lg 50 % embryonic infection doses (EID₅₀)] - A/Chicken/Kurgan/05/2005, A/Chicken/Crimea/04/2005, A/Chicken/Omsk/06, A/Chicken/Krasnodar/02/06, A/Chicken/Dagestan/06; strains with mid-level virulence [LD₅₀ (ALD₅₀) - 3,0-4,5 lg EID₅₀] - A/Duck/Kurgan/08/2005; and non-virulent strains [LD₅₀ (ALD₅₀) - >4,5 lg EID₅₀] - A/Turkey/Suzdalka/Nov-1/2005 and A/Chicken/Suzdalka/Nov-11/2005. Registered is the high degree of correlation (r = 0,89 ) between the indexes of virulence of the strains, obtained by means of intranasal and aerosol challenging of mice. The primary target-cells in mice, respiratory-infected by AIV, are the respiratory tract ciliated cells and pneumonocytes of I, II types.

Presented are the results of cytokines' (IL-1, IL-2, IFN-γ, and TNF-α, IL-10) level detection in blood sera of West Nile fever patients in the territory of Volgograd Region during the summer outbreak in 2010. Marked is a significant increase in IL-1, IFN-γ, IL-10 concentration and cytokine response prevalence of Th1-type.

Carried out are the studies of preparations of Francisella tularensis antigen complexes, obtained from the producer strains of different subspecies (holarctica, nearctica, mediasiatica, novicida). It is discovered that the outer-membrane (OM) antigen complex is a common antigen of Francisella tularensis regardless of its sub-specific origin. Antigen complexes isolated from strains of holarctica, nearctica, and mediasiatica subspecies have similar chemical and subunit composition, and immunobiological properties. Peculiarities of composition of F. tularensis subsp. novicida OM-antigen complex lead to the decrease of its immunogenicity and protective capability in comparison with analogous antigens of other subspecies.

Obtained are the immunomagnetic particles with the immobilized highly-specific monoclonal antibodies FB 11-x to Francisella tularensis. Their usage in enzyme-linked immunoassays made it possible to detect 10⁵ - 10⁶ CFU/ml of F. tularensis. The binding of the bacterial cells with IMP was confirmed by the results of fluorescent microscopy.

Carried out is approbation of the new nutrient media set for cholera diagnostics on the basis of bakers' yeast pancreatic digest within the frames of the special tactical training exercises for specialized anti-epidemic team (SAET). It is demonstrated that the developed nutrient media set compare favourably with its analogues, commonly used for practical procedures; and its constituent media have a potential to be included into the SAET mobilization reserve.

At CRIE, Moscow and RRAPI Microbe designed is the set of reagents for detection of Brucella spp. DNA in biological materials and cultures of microorganisms by means of PCR with hybridization-fluorescent registration of the results. This reagent set, AmpliSens ^® Brrucella spp.-FL, is supplied in two modifications versus the mechanism of the results registration (FRT - real time and FEP - end-point). High diagnostic value of the reagent set is determined in medical trials at L.A. Tarasevich Institute. The set is registered as medical device.

On the basis of silica - aluminosilicate, modified by carboxymethylated lignin and carbodiimide, obtained are the composite microgranulated magnetic immunoadsorbents (MIA) with high adsorption activity, which are characterized by the standardized structural characteristics and mechanical strength. Application of MIAs makes it possible, at the stage of tick samples preparation, to eliminate various admixtures via reiterative irrigations of the sorbent with the infectious agent fixed on it. Therefore negative influence of admixtures on the performed analysis is excluded, and the target agent is concentrated to the maximum limit. Thus the specificity and sensitivity of PCR-analysis enhances.

Presented are the data on the development and approbation of the method of Marburg, Ebola, and Lassa viruses identification based on real-time multiplex PCR with hybridization-fluorescent detection. This method is meant for the differential diagnostics of hemorrhagic fevers caused by these viruses. Displayed are the results of determination of multiplex PCR analytical sensitivity and specific activity.

Designed is the multi-analyte test for simultaneous immune chromatographic detection of A & B type botulinum neurotoxins (BT), using colloid gold nanoparticles. It is meant for food and environmental samples' analysis. The sensitivity of simultaneous BT detection of the A (30 ng/ml) and B (10 ng/ml) types is as high as that of mono-analytical tests, designed for one type BT detection. The test is demonstrated to be a specific one and can be used for BT detection in food stuffs.

Technology of O-antigen obtaining from Vibrio cholerae M41production strain, using the hollow fiber ultrafiltration modules, is developed and introduced into cholera vaccine manufacturing. Application of ultrafiltration makes it possible to obtain native high-activity O-antigen preparations which comply with the current regulatory requirements. Besides, it provides for economy (10-15-fold) of ammonium sulphate, used for antigen precipitation, reduces the duration of technological cycle (2 times), and the labor inputs. The technology of protective antigen concentrating is tested on V. cholerae strains MO45 and KM137 of O139 serogroup. Three series of experimental V. cholerae O139 vaccine are obtained.

Worked out is the experimental technology for protective antigens of Vibrio cholerae 569 B Inaba (cholera-anatoxin and O-antigen) concentrating by means of tangential ultrafiltration. Optimization of concentrating technological process is carried out. This technique makes it possible to obtain cholera vaccine components meeting all regulatory requirements.

Detected are several differences in biochemical properties of V. cholerae 569 B Inaba and V. cholerae M41 Ogawa productive strains. The differences are in the protein content in culture liquid, in the dynamics of enzyme activity in the process of reactor strain growth, and in the protease, phospholipase A2 and C activity. Demonstrated is the diverse resistance of enzymes of both of the strains to formaldehyde detoxification effect. Identified is the difference in enzyme activity of fractions, which are obtained by means of isoelectric precipitation of culture liquid proteins within the limits of pH 3,5-4,4.

Yersinia pestis recombinant strain KM277 (EV11MpFSK-3) Km^r is a potential producer-strain of capsular antigen (F1), since its productive properties and immunochemical activity of F1-antigen, synthesized by it, are higher than in Y. pestis EV and its derivatives. The apparent advantage of the Y. pestis KM277 strain is the ability to synthesize F1 antigen at 28 °C, and the absence of its own plague-microbe plasmids in cells. Worked out is the scheme of scaled cultivation of the recombinant strain and obtainment of capsular antigen preparation.

Demonstrated is the possibility of application of ultrafiltration technologies in the process of cholera toxin and plague agent capsular antigen precipitation under production conditions. Application of ultrafiltration techniques permits of the reduction of losses at the stages of isolation and purification of antigen preparations; and concentration of raw material or semi-finished product provides for the reduction of labor inputs. Thus it leads to the increase in productivity and economical efficiency.