This article provides the basic information on the current epidemiological situation on Zika fever across the world, the data on ZIKV global distribution, its main transmission routes, peculiarities of infection manifestations, and major approaches in the laboratory diagnostics. Taking into account the significant complication of the epidemiological situation on Zika fever in North, Central, and South America and the Caribbean, as well as the tendency towards the expansion of the areals of ZIKV effective vectors, identified are the key priority tasks to minimize epidemiological risks of Zika fever in the Russian Federation.

Objective of the study is to evaluate the state of parasitic systems and activity of natural plague foci of the Russian Federation in 2015 and to develop epizootiological forecast for 2016. In 2015, plague epizooties were detected in the territory of Gorno-Altaisk high-mountain, Tuva mountain, and Pre-Caspian sandy natural plague foci with a total area amounting to 1573.4 square kilometers. Isolated were 46 strains of plague microbe, including the isolates from rodents and leporines - 18, and from fleas - 28. Substantiated is epizootiological prognosis for sustaining challenging epidemiological situation in Gorno-Altaisk high-mountain and Tuva mountain natural foci in 2016. Demonstrated is the fact that consequently to effective prompt prophylactic measures (field deratization, disinsection) in 2015 decreased epizootic activity of Pre-Caspian sandy focus is observed. Specified is retention of low numbers of carriers and vectors of plague in natural foci of North and North-Western Caspian Sea Region, Pre-Caucasian, Caucasian, and Transbaikal Territories. Identified is upward trend for numbers of the little souslik in Pre-Caspian North-Western steppe, Volga-Ural steppe, Dagestan lowland-piedmont, and Pre-Caspian sandy natural plague foci.

In the course of assessment of epidemiological situation around the world in 2015, established has been the tendency to incidence rate reduction, with the annual average tempo - 10.225 % (in reference to 2006). For the first time ever, defined has been equivalent epidemiological year in case of cholera within an interval between 2014-2015.The key epidemiological risks of infection transmission globally are still inter-state (Africa, Asia) and extra-continental imports of cholera from the Caribbean (Cuba) and Asia into America (Canada) and Europe (Great Britain, Italy, Belgium, and Germany), which took place in 2015 too. Identified have been 21 endemic administrative territories in eight countries of Africa, America, the Caribbean, and Asia. Epidemiological complications are mainly caused by V. cholerae O1, biovar El Tor, producing cholera toxin of classical type, as well as by V. cholerae O1, classical biovar, and V. cholerae O139 serogroup. Epidemic manifestations of cholera in the Russian Federation are characterized by import of the infection by citizens returning from abroad; the isolation from surface water bodies of V. cholerae O1, biovar El Tor ctxA^- tcpA^-, ctxA^-tcpA⁺ and V. cholerae O139 ctxA^-and tcpA^-, as well as of single V. cholerae O1, biovar El Tor ctxA⁺tcpA⁺. Forecast on cholera world-wide and in Russia for 2016 remains unfavorable.

Analyzed are the data on epizootiological and epidemiological surveillance of tularemia foci, situated in the territory of 85 constituent entities of the Russian Federation. Positive findings, obtained in the process of examination of small mammals, Ixodidae ticks, mosquitoes, and environmental objects, using immunological and molecular-genetic methods; isolated tularemia agent cultures; as well as tularemia incidence rates among the population have made provisions for the assessment of circulation and infection activity in the Regions. In period of 2015, 71 cases of human infection with tularemia have been registered. Given is a brief characteristics of activity of the natural tularemia foci and epidemic situation in the territory of the Russian Federation in 2015. Specified are the regions where preventive vaccination is on a low level. Given is the differentiation of RF territories according to the risk of exposure to tularemia infection for 2016.

The intensity of epidemic West Nile fever (WNF) manifestations in the Russian Federation during the season, 2015 was low; morbidity rates coincided with those characteristic of inter-epidemic period. It total, 41 cases in 9 RF constituent entities were registered. WNF incidence rates among the population in the territory of Europe and North-American continent slightly exceeded morbidity rates, 2014, but then again did not reach epidemic levels. During the past two years seasonal development of WNF epidemic process in Russia has had a tendency to defer the incidence climax to early autumn. The presence of West Nile virus (WNV) markers was established in 11 entities of the Russian Federation in 2015, and existence of immunity to WNV in the population - in the 27 territories. According to the results of molecular-genetic surveillance, in the territory of the Russian Federation in 2015 circulation of WNV genotype 1a and 2 continued. The ongoing changes of climatic conditions towards warming predetermine high probability of further WNV circulation in the environment and emergence of infection cases in areas of the country far to the north. However probability of WNF epidemic manifestations intensification in 2016 remains insignificant.

Analyzed has been epidemiological situation and measures, performed for prophylaxis of tick-borne viral encephalitis in the territory of Russia in 2015. It is shown that the number of humans bitten by ticks increased in the majority of the constituent entities of the country. But specific and nonspecific preventive operations in 2015 were realized to a lesser extent as compared to 2014. Along with the natural factors, it might be the reason for increase in human tick-borne viral encephalitis morbidity rates. In total, 2116 patients with tick-borne viral encephalitis and 24 lethal cases were registered in the country. On the basis of the data regarding tick-borne viral encephalitis (TBVE) incidence rate among the population across the Federal Districts of Russia over a period of 2009-2015, forecasted have been intensive indicators of the clinical forms’ manifestations for 2016. TBVE morbidity rate in RF will amount to (1.90 ± 0.21)^о/_оооо. With 95 % probability it will be retained within a range of 1.4-2.4^о/_оооо.

This paper presents analysis of epidemiological situation on Crimean-Congo hemorrhagic fever (CCHF) in Russia in 2015; summarized are the results of epidemiological survey of the territory of the natural CCHF focus in the south of the European part of Russia. In 2015, the Russian Federation reported 139 cases of CCHF. The most significant increase in the incidence of CCHF occurred in the Stavropol Territory and Rostov Region. Identified was CCHF case in Voronezh city imported from the Crimean Federal District. Marked is the involvement of new administrative areas in the epidemic core of the natural focus. The aggravation of the epidemiological situation in the South of Russia was mainly due to the favorable for dissemination of ticks, H. marginatum, climatic conditions in winter 2014-2015. In 2016, in the case of successful Ixodids wintering, as well as ineffectively exercised acaricide treatments, the numbers of H. marginatum may increase compared to 2015, which may become the cause of augmented incidence among the population.

Given are the data on the circulation of highly pathogenic flu virus, A/H5, within the last two years. Discussed is the current state of the situation on H5 flu in the territory of the Russian Federation, where during 2014-2015, for the first time since 2010, registered has been the isolation of highly pathogenic flu virus, H5N1 and H5N8 subtypes. It is demonstrated that the territory of Russia plays a significant role in trans-continental transfer of flu viruses by wild birds from South-Eastern Asia into Europe and North America. Moreover, an assumption is made that a continued circulation of highly pathogenic viruses in the territory of the country is quiet possible.

Objective of the study is to characterize the case of human plague in the territory of Gorno-Altaisk high-mountain natural focus in 2015 and to analyze the initiated measures, associated with localization and elimination of epidemic focus. Materials and methods. Utilized are the data contained in reporting and source (primary) documentation of the FGHI “Altai Plague Control Station”, records of the Rospotrebnadzor Administration in the Republic of Altai, and information collected by the FGHI “Irkutsk Research Anti-Plague Institute”. Results and conclusions. Human infection occurred as a result of gray marmot dressing, which was caught in the Elangash River-Valley, against the background of unfavorable epizootic situation, caused by proliferation of the plague agent of main subspecie in the territory of the focus. Clinical material investigations, performed by means of bacteriological and molecular-genetic methods, showed negative findings. Applying serological method within the system of indirect hemagglutination test (IHAT)/antigen neutralization test (ANT), in blood sera, obtained at the time of hospitalization, low-titred specific antibodies to plague microbe (recognized as post-vaccinal ones) were detected. In blood serum sample, obtained 7 days later, identified were high-titer antibodies, which allowed for confirmation of clinical diagnosis - “bubonic plague”. Due to efficient cooperation between Rospotrebnadzor institutions and medical facilities, as well as municipal authorities it was possible to avoid further development of anthropogenic transmission of plague; to localize and eliminate epidemic outbreak of this dangerous infectious disease in Kosh-Agach Region of the Republic of Altai in the shortest possible time.

Objective of the study was to investigate a criminal case of infection with HIV, applying molecular-genetic analysis of blood plasma samples from an estimated source of an infection and the recipient for evaluation of probability of epidemiological connection between them. Materials and methods. The study involved genotyping and phylogenetic analysis of nucleotide sequences of HIV-1 variants, isolated from patients in the investigated group and the control one (19 nucleotide sequences of the HIV-1 from the patients living in the Saratov region, and 15 nucleotide sequences from GenBank). Genotyping was performed using the commercial ViroSeq HIV-1 Genotyping System. The sub-typing of HIV-1 strains was carried out on-line, through the COMET HIV-1/2 and HCV and REGA HIV-1 Sybtyping Tool programs. Phylogenetic analysis of nucleotide sequences was carried out by Mega software, version 5.2. Phylogenetic trees were constructed; nucleotide distances were calculated by Kimura method (bootstrap level 1000). Results and conclusions. Virus variants, isolated from the studied samples, were defined as HIV-1 A subtype. Performed phylogenetic analysis showed that nucleotide sequences of the studied samples authentically grouped on the phylogenetic tree, forming a common cluster, which mismatched that of control group. Calculation of the genetic distance testifies that the genetic relation between the samples within the investigated group is higher, than between the same samples and those of the control group. Thus, by means of phylogenetic analysis it is shown that the strains received from an estimated source of infection and the recipient are genetically closer to each other, than to the strains from the group of comparison. In this regard, it is possible to claim with a big share of confidence that probability of epidemiological connection between them exists.

Objective of the study is to demonstrate technical and operational characteristics of the specialized suit, its protective efficacy in relation to respiratory organs and skin at different air-injection rates through analyzing the possibility to utilize individual protective equipment, connected to pneumatic line, from the viewpoint of biological safety. Materials and methods. Tested has been specialized suit of a combined type. Investigations have been conducted in an aerosol cabinet, involving volunteer researches, who performed controlled medium physical activities within 4 hours term. In the course of the studies analyzed have been the following physical parameters of the test operators: heart rate, respiratory rhythm, body temperature. Identification of layer-by-layer contamination of the specialized suit (external and internal surfaces), linen and skin of the volunteers has been carried out applying washing techniques and triple agar prints. Results and conclusions. Testing of the designed suit has revealed that its utilization provides for proper solid protection of healthcare personnel from microbial aerosol, and adherence to desired speed of air-injection into interlayer (under-suit) space, ≥ 250 l/min, allows for maintaining high working capacities.

Iron is an essential for growth and reproduction of bacteria element. One of the ways of its acquisition by Bacillus anthracis is utilization of high-affinity chelating agents of iron ions - bacillibactin and petrobactin siderophores, extracting the iron from transferring and ferritin of a host cell. Bacillibactin and petrobactin functions are realized on different stages of B. anthracis growth in vivo, whereas petrobactin synthesis is also necessary for manifestation of microbe virulence. Awareness of siderophore biosynthesis pathways facilitates the development of medicines against anthrax, which can block them up. The review contains the data on the structure, genetics, and functions of B. anthracis siderophores.

Objective of the study is to investigate the sensitivity of different animals to highly pathogenic Orthopoxviruses applying techniques, based on utilization of primary cultures of lung cells, and to assess the possibility of further deployment of this approach. Materials and methods. Cultural and virological research methods are used. Results and conclusions. Performed is the assessment of sensitivity of outbred mice, marmots and chickens to variola virus (VV) and monkeypox virus (MPV), using suspended primary cultures of lung cells (SPCLC) of these animals. Through inoculation of the mentioned above cell cultures with VV and MPV in a dose of 0.00001 PFU per a cell (plaque forming unit /cell) demonstrated has been virus replication with maximum concentration values in all cases (1,4 - 2,0 lg PFU/ml), mainly 3 days after infection. According to the data on SPCLC, sensitivity to VV in mice, marmots and chickens (ID₅₀ - 50 % infective dose) amounts to (1,3 ± 0,5) lg PFU; (2,3 ± 0,5) lg PFU; and (0,0 ± 0,4) lg PFU respectively, taking into account unhindered interaction of the virus with permissive lung cells in the organism of the animals. As for MPV values for this indicator, they are: (1,7 ± 0,3) lg PFU for mice, and (0,5 ± 0,3) lg PFU - for marmots. Obtained ID₅₀ values for VV using mice SPCLC and for MPV using mice and marmots SPCLC coincide with the ones, studied in direct experiments on intranasal infection with the viruses, with regard to 10 % of the viral application in lungs when deploying the latter method of infection. The fact testifies to the possibility of further deployment of this method for the assessment of animal sensitivity to highly pathogenic Orthopoxviruses based on the results of in vitro experiments.

Objective of the study is to optimize the algorithm for authenticity specification of pathogenic bacteria strains and to evaluate its efficacy for nomenclature examination of the isolates, Bacillus genus, from the State Collection of Pathogenic Bacteria functioning at the premises of RusRAPI “Microbe”, the authenticity of which bore scrutiny. Materials and methods. Methodological approach is based on application of automated microorganism identification systems: bacteriological analyzer Vitek 2, automatic ribotyping station DuPont Qualicon RiboPrinter System, and mass-spectrometer MicroFlex MALDI Biotyper, followed by complex assessment of the results obtained, using BioNumerics 7.1 software product. Results and conclusions. It is established that automated analyzers perform reasonably accurate identification of the investigated microorganisms in reference to their specie appurtenance. Vitek 2 shows the best efficiency. Unlike other analyzers, it allows for differentiation of B. anthracis STI-1 strain from a group of bacilli, B. cereus specie. It is of note that different systems range B. megaterium 5 strain in different ways. Carried out complex analysis of the results, obtained from all the three automated devices, using BioNumerics 7.1 software, relegates B. megaterium 5 to B. licheniformis with a high degree probability. Thus, it is necessary to include in the algorithm several techniques with subsequent complex analysis of the data obtained to specify authenticity and taxonomic appurtenance of the collection strains under nomenclature examination.

Objective of the investigation was to model the adverse action of vaccinia virus (VV), caused by oral immunization of mice and to evaluate efficacy of its reduction, using therapeutic and prophylactic drugs. Materials and methods. Virological and immunological research methods were used. Results and conclusions. Reproduced was pathological action of VV in the orally infected mice. The ability to reduce the side effect and protect mice from lethal infection was demonstrated by such preparations as Metisazon, Likopid, and NIOCH-14 orally administered in the investigated schemes. Moreover preliminary single oral immunization with TEOVak smallpox vaccine before oral infection with Neurovaccine-92 strain of VV also lowered pathogenic effect and protected mice against death. All the investigated schemes of drug administration did not affect the immune response if used alongside with TEOVak smallpox vaccine and can be deployed to develop safe schemes of primary oral vaccination against smallpox. In addition, such drugs as Ribomunil, Immudon, Ingavirin can be used as means to enhance the immune response to smallpox vaccines.

Objective of the study was to substantiate experimentally the possibility to enhance efficiency of manufacturing process through application of novel engineering solutions in the course of freeze-drying of vaccine immunogens. Materials and methods. Choleragen-anatoxin and O-antigens, Inaba and Ogawa, served as specific immunogenic components of the vaccine. Identified were temperature values for O-antigen eutectics (approx. -35 ºC). At reaching -55 ºC choleragen-anatoxin froze incompletely (fractionally). Full solidification of the preparation was achieved by means of the “annealing”. Revealed was non-effect of the tested freeze modes on the properties of antigen components of chemical cholera vaccine. Desired values of technological parameters for primary drying and desorption were specified by trial. Investigations of activity of chemical cholera vaccine immunogen lyophilizates testified to the compliance with the rated critical requirements. Results and conclusions. Results of the study provided for significant time expenditure reduction and obtainment of high-quality preparations at once.

Objective of the study is to experimentally substantiate the possibility to improve manufacturing efficiency by means of mass reduction of a vaccine tablet from 300 to 100 mg. Materials and methods. Inaba O-antigen lyophilizate serves as the specific immunogenic component of the vaccine. Results and conclusions. It is identified that it is expedient to produce tablets of 6 mm in diameter. Justified is the quantitative content of additive substances (lactose monohydrate, micro-crystal cellulose, and polyvinylpyrolidone). Moreover, the studies have specified target values for technological parameters of such processes as fluid bed granulation of the formula with overfeed of the binder, tablet compression and enteric-coating (Acryl-eze) application to finished dosage form. Using Inaba O-antigen lyophilizate manufactured has been model experimental series of the vaccine. Investigated have been its characteristics. Verified vaccine quality indicators testify to the compliance of the product with the requirements of manufacturer’s pharmacopoeial monograph. The studies exercised showed the possibility in principle to enhance manufacturing efficacy through the decrement of additives amounts, and thus the mass of a vaccine tablet from 300 up to 100 mg.

Objective of the study is to deploy 2-D electrophoresis for the construction of lysate “protein profiles” of bacterial cultures of particularly dangerous infections. Materials and methods. “Protein profiles” are obtained on the model of non-toxigenic V. cholerae biovar El Tor M-888 strain and Y. pestis EV NIIEG strain, cultivated at 28 and 37 °C. Commercial sample of E. coli strain lysate serves as comparator (reference) product. SDS-PAGE-electrophoresis is carried out following classical Laemmli approach. 2D-electrophoresis is based on O’Farrel technique and manufacturer’s recommendations. For protein visualization, staining of electrophoregrams with coomassie blue G-250 and silver nitrate is used. Protein concentration (load) is evaluated with the help of Bradford protein assay. Cell lysate samples are purged of non-protein impurities applying Ready Prep 2-D Cleanup Kit. Results and conclusions. Described is a highly effective complex method of protein mixture separation, which assumes preparation of bacterial culture probes where cells are fractured under ultrasonic exposure in the lysing buffer, then purified and analyzed either in reference to the charge in pH gradient, or - to molecular mass in SDS-PAGE-electrophoresis. After the staining 2-D gels are assayed by means of “Dymension” software product, installed on the platform of multi-functional gel documentation system “Syngene”. Utilization of stripes with immobilized pH gradient of a preset range, commercial reagent kits, and large-sized polyacrylamide gels has allowed for optimization of the conditions for performing 2D-electrophoresis, reduction of the time elapsed, and improvement of accuracy and reproducibility. On the model of non-toxigenic V. cholerae biovar El Tor M-888 strain and Y. pestis EV NIIEG strain, cultivated at 28 and 37 °C, applying 2D-electrophoresis, obtained are the protein “profiles” of the investigated strains, demonstrated is high efficacy of the method and possibility of its deployment for studies of gene expression dynamics as regards agents of particularly dangerous infections.

Nowadays, problem of tableted drug form contamination with extraneous micro-flora is in the spotlight of scientists, as the specific share of these medical preparations in the world market amounts to more than 60 % and has a tendency to increase. Thus, objective of the study is to investigate the degree of contamination of the basic and auxiliary raw materials at different stages of live plague vaccine manufacturing, rapid dissolving tablets, and the ways to reduce it. Materials and methods. Utilized has been lyophilized Y. pestis live culture of the vaccine strain EV NIIEG, and the live plague vaccine, rapid dissolving tablets. Carried out has been assessment of “microbiological purity” at different stages of tableted live plague vaccine manufacturing: grinding, mixing, granulation, sublimation, and palletizing. Results and conclusions. Identified is the dynamic pattern of quantitative micro-flora composition of the mentioned above drug. Established is the alteration of microbial impurity at separate technological manufacturing steps. Specified is the technological stage with the most expressed contamination. Analysis of factors, which affect vaccine impurity, has revealed that finished dosage-form quality improvement is impossible without incoming control of stock and auxiliary materials, as well as enhancement of manufacturing procedure up to the level, complying with applicable pharmaceutical production standards. It is experimentally proved that series of tableted live plague vaccine, obtained using modernized technological equipment, provide for 3-fold reduction of contamination.