New area of biological studies that goes under general name of “synthetic biology” is a next step in the development of gene engineering associated with design and construction of unique biological systems with “preset” functions and properties, having no natural analogues. Nowadays synthetic biology is a source of innovations that offer solution to a number of global problems facing the humanity, including production of artificial genome-based multi-diagnostic panels, medicinal preparations, synthetic vaccine, etc. The process of unnatural life form creation requires conduction of monitoring both on the international and national scales using advanced system of biological risk assessment.

Objective of the work was to carry out complex assessment of the current state of epizootic activity and epidemiological significance of the infectious disease natural foci in the Aleksandrovsk-Sakhalin territory of the Sakhalin Region. Trapped were 56 samples of small mammals in July, 2010; collected were 180 specimens of taiga tick imago, caught were 1000 specimens of mosquitoes. 223 samples of blood sera were taken from residents of the region. All the field data were tested to detect specific antibodies, antigens and genetic material of agents. Based on the results of epizootiological investigations, serological and molecular-genetic assays, demonstrated was the occurence of natural foci of leptospirosis, tularemia, tick-borne encephalitis, borreliosis, rickettsiosis, human granulocytic anaplasmosis, human monocytic ehrlichiosis, West Nile fever, Inco fever, Batai and Geto fevers, as well as HFRS with varying degree of activity manifestation in the territory of the region. Isolated was tick-borne encephalitis virus from mosquitoes.

Represented is the analysis of epidemiological situation on vector-borne natural-focal infections in the specially protected eco-resort territory of Russia - Caucasian Mineral Waters of the Stavropol Region. Climactic peculiarities of Caucasian Mineral Waters with their landscape diversity, high abundance rates of ticks and mosquitoes, and anthropogenic impact on ecosystems create favorable conditions for natural foci formation. Crucial epidemiological significance in the regional infectious pathology is attributed to tick-borne borreliosis. Registration of Crimean hemorrhagic fever cases, as well as West Nile (retrospectively) fever and tularemia cases, and identification of specific antibodies to etiological agents of these infections in blood sera of donors testify to volatile epidemiological situation on vector-borne natural-focal infections in the region, which means that there is a need for further ecological, epidemiological and epizootiological monitoring as a constituent element of epidemiological surveillance aimed at provision of sanitary-epidemiological welfare of the population.

Within the period of 2000-2011, in the West of Kazakhstan, identified have been five, previously unknown in the territory, natural foci of dangerous infectious diseases such as hemorrhagic fever with renal syndrome, Astrakhan spotty fever, Crimean hemorrhagic fever, West Nile fever, and tick-borne viral encephalitis. The reason is that key ecological factors for the persistence of the infections in the local biocoenoses occurred. It is characteristic that circulation of the agents of new infectious diseases is registered in the territory of the long-established natural plague and tularemia foci. Since 2000 and on, hemorrhagic fever with renal syndrome morbidity is registered in the Western-Kazakhstan Region on a regular basis. There is some evidence to identification of West Nile fever patients in the territory of Russia in 2012, which came from Kazakhstan. Based on the spatial distribution of the natural foci of various infections, Western-Kazakhstan Region has been subdivided into four areas. In order to provide for the effective prophylaxis of emerging diseases, it is essential that healthcare facilities and services dealing with infectious diseases are consolidated and reinforced, and epidemiological surveillance is improved with the current conditions in mind.

A set of documents concerning biological safety at the institutional level is prepared and put in to practice. Three levels of specialists` admission for works with pathogenic biological agents of the I-IV groups have been developed, substantiated and introduced. Considered are the variants of execution of coupled (simultaneous) work with PBA of different groups. The statement on control of disinfectants received by the institution defines the order of control execution as regards newly received disinfectants and correspondence of disinfectants` working solutions in the laboratories to the requirements of sanitary regulations. Demonstrated is practicability of the assessment of protective efficacy of air-cleaning filters in the exhaust ventilation systems of “contaminated” area rooms in the laboratories carrying out the works with PBA using bacterial aerosol and physical method.

Epizootiological monitoring of the forest-steppe area of the natural tularemia focus in the Stavropol region has revealed that the role of particular species of small mammals in the persistence of natural tularemia focality is unequal. Epizootic activity of the focus in 1959-1970 was determined by the numerous species of rodents: Microtus arvalis , mice of Syvaemus genus and Mus musculus . In 1972-2010 there occurred significant changes in the grouping of the main tularemia agent carriers under the influence of strong anthropogenic pressure. Nowadays the leading role is played by the widely-spread and subsistent mice of Sylvaemus genus and C. suaveolens , the latter ones being responsible for 31.2 % of overall, isolated from small mammals, tularemia agent strains. In addition to this, epizootic significance of M. arvalis has greatly changed. Index of strains isolated from field vole has lowered from 55.3 up to 28.4. Numbers of M. arvalis and Mus musculus are continuously on the low level, which is due to the absence of favorable breeding conditions. It reduces their impact on the persistence of natural focality in the territory under surveillance significantly.

In recent years, an increase of tourist flow, strengthening of social and economic connections between the countries take place. The processes of the world economy globalization, development of international transport systems, and migration of population form preconditions for the spread of new infectious diseases in the territory of Russia and CIS countries. Imported cases of blastomycosis and paracoccidioidomycosis are already registered in many countries of Europe and Asia among persons who returned from endemic countries. Thus, it is likely to find infected persons and patients in Russia. The purpose of our review is to provide data indicating the need for improvement of methods for epidemiological control of blastomycosis and paracoccidioidomycosis. Diagnostic studies should be carried out by competent personnel in specialized laboratories that meet the necessary safety requirements. Thus, awareness about these diseases of as many experts as possible, guarantees the successful implementation of strategies for diagnostics and treatment.

Objective of the work was to develop selection criteria for the dummy Y. pestis strains, as well as principles of the setting-up a panel and its application for practicing laboratory and differential diagnostics of plague. Studied were the RF regulations, statutory documents and methodological recommendations on the laboratory diagnostics of plague and safety of works with microorganisms; training courses for specialists to qualify for work with the agents of particularly dangerous infections. Research method: analytical. Consequently, established were the term for “dummy strain”; selection criteria for the Y. pestis strains used in the practical course within the frames of the training programme “Microbiology and Laboratory Diagnosis of Plague”; and algorithm of the course application in view of biological risk mitigation during the process of education.

Consequently to the long-term investigations conducted in natural and anthropourgic foci of plague, situated in Africa, Eurasia, North and South America, identified were not less than 340 species of mammals and birds infected with plague agent. The paper contains the list of plague agent carriers described, both in domestic and foreign publications.

Concerned are the issues related to enterobacteria omptins, their structure and functionality, as well as alternative role in pathogenesis of the infections induced by them. Isolated from cholera vibrio, and later purified using differential centrifugation and column chromatography has been porin protein of the OmpT outer membranes, with the molar mass of approximately 40 kDa. Synthesis of porin is under control of the complex regulatory system. It does not contain cysteine, but possesses proteolytic activity with broad substrate specificity: it hydrolyzes fibrin, protamin, gelatine; transduces human plasminogen into plasmin, which provides for the well-known advantages for the vibrios in the intestine of a susceptible host. Comparative computer-assisted analysis of amino acid sequence has revealed that cholera vibrio OmpT protein relates to the omptins of enterobacteria as a far-remotely one, and has 13 % identity and similarity to it. OmpT protein is probably affiliated to a new class of porins of the family Vibrionaceae .

A set of primers for detection of genes encoding resistance to streptomycin ( strA, strB ), tetracyclin ( tetA, tetR ), chloramphenicol ( catА ), kanamycin ( npt , aphA ), vankomycin ( sanA ), polymyxin ( pmrD ) has been developed with the aim of rapid and effective detection of drug-resistant strains of dangerous bacterial infections agents. Efficacy of constructed primers has been confirmed against a panel of 40 Yersinia pestis, 49 Vibrio cholerae, and 2 Escherichia coli strains from the State collection of pathogenic bacteria of the RAPI “Microbe”. Drug-resistance genes ntp and catA have been detected in plague agent strains , strA, strB , npt , aphA , tetA and tetR - in cholera agent; strA , tetR , ntp and aphA - in pathogenic strain E. coli О157:H7. Determined is universal character of the designed primers for drug-resistance genes detection in these pathogenic bacteria species.

Anniversary of melioidosis agent discovery. Discussed are the key advances of world science in the sphere of Burkholderia pseudomallei ecology, epidemiology, clinical picture, and treatment of the disease. Apart from this, summarized are the data on the 50-years long study of melioidosis, carried out by the specialists of Plague Control Agency of the Russian Federation.

By the example of Martin Christ Epsilon 2-6D device carried out was assessment of the possibility to use freeze-dryers of the chamber type for conservation of pathogenic microorganisms collection strains. Elaborated was algorithm of lyophilisation of the III-IV pathogenicity groups bacteria, which incorporated conditions of freeze-drying and biological safety provision of this process. Indices of viability and survivability were defined for freeze-dried cells of pathogenic bacteria strains. Using thermostability test calculated were predicted timelines of storage of collection strains preparations freeze-dried in the flasks in Martin Christ Epsilon 2-6D. It was determined that in the collections of pathogenic microorganisms freeze-dryers of the chamber type could be used most prospectively for the III-IV pathogenicity groups bacteria conservation requiring mass reproduction and not intended for long storage. At the same time their application for freeze-drying of the strains of the I-II pathogenicity groups bacteria intended for a long storage, requires further adaptation of these devices as regards biological safety provision and prolongation of the shelf life.

CFU/ml) and soluble antigens (FI) − ≥ 4.8 ng/ml and high specificity, confirmed by the absence of false-positive reactions with five heterologous microorganisms. The test-systems were used for Y. pestis antigen detection in field material from the territory of the Altai mountain natural plague focus by dot-immunoassay with comparison of the received results in passive hemagglutination reaction. Test-systems possessed a number of advantages as compared to routine serological reactions and could be applied with success by practical public health services both in stationary and field conditions.

Considered are the stages of the SAET laboratory facilities development. Initially main area of activities was assumed to be specific indication of bacterial threats and laboratory control over the ambient environment objects for the presence of particularly dangerous infectious disease agents. Significant increment in the workload occurred during the period of localization and elimination of epidemic cholera manifestations in the 1970s, when primary tasks of bacteriological unit consisted in carrying out mass bacteriological investigations of samples from humans, environment objects, and food items. Assignment of new functions to the laboratory facilities, such as performance of sanitary-microbiological investigations, monitoring over ambient environment objects for the presence of vibrio-flora and natural-focal infectious disease agents, clinical material assays - is associated with SAETs participation in liquidation of medical-sanitary consequences of natural disasters and human cost relief as aftermaths of military conflicts, as well as participation in management of mass events with international representation. Most important issues in the development of SAET laboratory facilities are implementation of advanced diagnostic technologies, automatization of various stages in the process of analysis performing, standardization of diagnostic investigations, and ensuring compliance of the facilities with national and international requirements.

Mass spectrometry is a modern physical-chemical analytical method that provides for qualitative and quantitative assessment of the substance composition. It is based on pre-ionization of the atoms and molecules included into it. One of the advanced methods of ionization, due to which mass-spectrometry investigation of macromolecules has become a frequent practice, is matrix-assisted laser desorption/ionization (MALDI). The essence of it is the pulsed laser irradiation of the matter under study, mixed with the matrix. The review discusses current data on MALDI mass-spectrometry application for the performance of species-specific and genus-specific identification of microorganisms at the premises of diagnostic laboratories. Considered are the basic advantages of MALDI-TOF identification as compared to bacteriologic, immunologic, and molecular-genetic methods of assessment. Allocated is the mass-spectrometry position in the system of laboratory diagnostics of infectious diseases, including particularly dangerous ones, in the territory of the Russian Federation.

B. pseudomallei 100 resistance (61.5 % of survived animals). It was concluded that it is a good practice to include bestim in the schedule of complex immunization against melioidosis.

The members of the Filoviridae (Marburg and Ebola viruses) and Arenaviridae (Lassa, Lujo, Machupo, Junin, Guanarito, Sabia viruses) families are the etiological agents of particularly dangerous viral hemorrhagic fevers. These agents pose a potential threat to public health care in view of the possibility of their unintended import into the non-endemic regions, and thus construction of specific medical protectors as regards induced by them diseases is a pressing issue. According to leading experts, vaccination of the cohorts that fall in the risk groups is the most effective and least expensive method to prevent the development of epidemics. The review contains information on a new prospective line of protective preparations development as regards particularly dangerous viral infections - construction of alphavirus-replicon-based vaccine. Elaboration of recombinant replicons does not require cultivation of pathogenic microorganisms. RNA-replicons are distinguished by their incapacity to produce infective progeny, which is of a great importance for the development of vaccines against particularly dangerous viral hemorrhagic fevers. Advantages of alphaviral replicons over other RNA-replicons are as follows: high levels of heterologous gene expression and resistance to anti-vector immunity. RNA-replicons of alphaviruses combine the safety of inactivated, and immunogenicity of live attenuated vaccines. Alphaviruses-based replicons are suitable for express vaccine development with the purpose of specific prophylaxis of viral infectious diseases.

Objective of the work is to carry out comparative assessment of the pancreatic hydrolysates of protein-containing products, both phytogenous and zoogenous, as nutrient base for microbiological media. Gelatine, soy, soy concentrate, maize gluten, fish meal, common kilka, and bovine blood have been used as a feedstock. Protein stuff hydrolysis, hydrolysate purification, and validation of physical-chemical properties were performed in accordance with conventional techniques. Testing of peptone biological parameters has been carried out on the model of nutrient agar using Shigella flexneri 1a 8516, Shigella sonnei “S form”, Pseudomonas aeruginosa 27/99, Serratia plymuthica 1 test strains. Identified have been physical-chemical parameters of the hydrolysates under study. Detected are the variations in quantity, diameter and frequency of dissociation among the colonies of Shigella flexneri 1a 8516, Shigella sonnei “S form”, chromogenesis in Pseudomonas aeruginosa 27/99, Serratia plymuthica 1, cultivated on agar media with hydrolysates under study. Obtained are the comparative data on physical-chemical and biological parameters of all experimental hydrolysates, which offers an opportunity to differentiate their choice when adding them into bacteriological nutrient media.

Presented is mathematical model of kinetics of the process of O-antigen and cholera toxin synthesis during periodical submerged cultivation of V. cholerae 569В Inaba with limitation as regards carbonic substrate. The proposed model is based upon analysis of experimental data on V. cholerae 569В Inaba biomass and antigens accumulation, rate of growth and antigens release, and glucose utilization. Using Mathcad 15.0 software calculated are coefficients of differential equations entering into the mathematical model. Comparison of predicted and experimental data demonstrates that relative error of determination of concentrations of the synthesized substances, glucose and cholera vibrio is between 5 and 20 %. The proposed model permits to determine maximum output of final products and specify the parameters of cultivation process performance at different initial conditions.

Put forward are two alternatives of a new method for optimization of conditions of LPS obtaining and purification from Y. pestis strains; as well as for avoiding application of poisonous and hard-to-remove reagents; for simplification and cost-cutting of the technique; and for rationalization of production waste management. This method involves preliminary salt-water extraction of bacteria, for elimination of easy-dissolving substances, with the subsequent fracturing using ultrasound in lysing buffer (0,1 M Tris-HCl, pH 8,0; 10 mmol of EDTA, 1 % Triton X-100). The first alternative for deproteinization of non-purified endotoxin is the commercial preparation of proteinase K (Sigma), the second one - an enzyme complex - proteovibrin, isolated from waste material accumulated in the process of cholera chemical bivalent vaccine production. Apart from this, introduced has been a phase of sample acidification by applying glacial acetic acid up to pH 3,2-3,4 to decontaminate LPS from nucleic acids. These two variations of the method provide for enhancement of LPS preparation quality as compared to prototype method, and for obtainment of plague agent endotoxin that is hardly distinguishable in physical-chemical properties, homogeneity, immunochemical activity and specificity from the antigen, manufactured by means of water-phenol extraction following Westphal O. technique.