[The comparative evaluation of effectiveness of laboratory techniques of diagnostic of anthrax and detection its agent in objects of environment].

The analysis of samples received during ictus of anthrax in the Stavropolskii kraii in 2013 permitted to study comparative effectiveness of regulated methods of laboratory diagnostic. The effectiveness of bacteriological, biological and molecular methods and necessity of their complex application for receiving optimal results are confirmed. The rapidity, effectiveness and specificity of polymerase chain reaction is emphasized. This method in case of absence of isolation of anthrax microbe can be the only method of confirming diagnose in people in aggregate with typical clinical picture and corresponding epidemic situation.

[EPIZOOTOLOGICAL AND EPIDEMIOLOGICAL ZONING OF THE TERRITORY OF NORTH CAUCASUS FEDERAL REGION BYA DEGREE OF NON-WELFARE BYANTHRAX USING ARC GIS10 PROGRAM].

AIM: Conducting epizootological and epidemiological zoning of the territory ofNorth Caucasus -Federal Region (NCFR) by the degree of non-welfare by anthrax using Arc GIS10 program. MATERIALS AND METHODS: 2 parameters were used during zoning ofthe territories of subjects of NCFR (Stavropol Region, Karachaevo-Cherkessia, Kabardino-Balkaria, Chechen, North Ossetia-Alania, Ingushetia, Dagestan Republics) by non-welfare by anthrax: integrated parameter of infection risk and epizootics risk. The parameters were input into the Arc GIS 10 program. Materials on infection of humans and animals with anthrax for the last 55 (1960 - 2014) years were used. RESULTS: The regions of the territories of NCFR subjects were distributed by non-welfare degree into 4 groups: group 1 - regions with a low degree of non-welfare, group 2 - regions with an average degree of non-welfare, group 3 - regions with a high degree of non-welfare, group 4 - regions with a very high degree-of non-welfare. CONCLUSION: Taking into account high parameters of spread and inten- sity of epizootological and epidemiological situation, regions of groups 3 and 4 could be classified as endemic territories for anthrax, whereas all the other regions could be attributed to the zone of sporadic infection manifestation.

[Use of GIS technologies in development of a cadaster of stationary unfavorable by anthrax points in Krasnodar region].

AIM: Development of a cadaster of stationary unfavorable by anthrax points in Krasnodar region by using GIS technologies. MATERIALS AND METHODS: Ageographical information system (GIS) of ESRI program--Arc GIS 10.1 was used for the composition of an electronic cadaster of stationary unfavorable by anthrax points. Topographic basis--electronic maps of Krasnodar region (M 1:200 000). RESULTS: An evaluation of safety by anthrax of Krasnodar region areas bythe amount of stationary unfavorable by anthrax points situated in its territories, by their activity during the 76 year observation period was given. Spatial adjoint analysis of ecological timing of stationary unfavorable by anthrax points to soil and landscape zones in Krasnodar region was carried out. CONCLUSION: Electronic cadaster of stationary unfavorable by anthrax points in Krasnodar region may serve as a basis for purposeful implementation of prophylaxis measures.

[Analysis of outbreak of anthrax in Omsk region in 2010].

AIM: Carrying out analysis of epizootologic-epidemiologic situation on anthrax that had emerged in Omsk region in 2010 when horse meat from epizootic focus of anthrax was used in production of meat semi-finished products. MATERIALS AND METHODS: Study of samples for detection of anthrax causative agents and strain identification was performed according to guidelines 1.3.2569-09. Strain genotyping was performed by MLVA method. RESULTS: The epizootologic-epidemiologic investigation performed allowed to detect the causes of emergence of anthrax outbreak, its routes and factors of transmission. MLVA genotyping results gave evidence on the single origin of Bacillus anthracis strains isolated from sick animals, humans and food substances. CONCLUSION: Timely execution of a complex of epizootic and epidemic control measures allowed to localize epizootic and epidemic focus of anthrax as well as prevent a possible large scale development of epidemic complications.

[Genotype diversity of the Bacillus anthracis strains isolated from the Caucasus region].

The study of the genotypes of Bacillus anthracis strains isolated from the Caucasus region was performed using MLVA. Among 149 strains, 32 distinctive MLVA-8 genotypes belonging to Ala, A3a, A4 and B1 molecular diversity groups were identified. 9 genotypes were not described previously; 6 genotypes were not found in other geographic regions and could be considered as endemic for Caucasus. The majority of the identified genotypes are widespread not only in Caucasus, but also throughout Eurasia, Africa, and America. Molecular diversity of Caucasian isolates is comparable to the worldwide diversity. This represents historical relations of this region, proximity to ancient trade routes and intensity of the anthrax epizootic in Caucasus. The obtained results are of interest from the theoretical point of view, as well as for the application in epidemiological research of the anthrax outbreaks.

[Methodic approaches to testing Bacillus anthracis susceptibility to antibacterials].

Susceptibility of 50 isolates of Bacillus anthracis to 24 antibiotics was tested by the disk-diffusion method and the method of serial dilutions in solid media. The tests allowed to determine the boundary values of the growth inhibition zones and the minimum inhibitory concentrations of the antibiotics for susceptible and resistant strains of B. anthracis. Nutrient media and reference strains for testing antibiotic susceptibility of B. anthracis are recommended.

[Phenotypic and genetic features of cultural-morphologic variants of Bacillus anthracis].

Comparative analysis of MVLA-genotypes of 6 Bacillus anthracis strains and 40 their variants differing on capsule- and toxin synthesis, hemolytic, proteolytic and lecitinase activity, nutritional requirements, susceptibility to anthrax bacteriophages, virulence, immunogenicity, and presence of genes for capsule and toxin synthesis was performed. Results of phylogenetic analysis of 5 chromosome locuses and plasmid locus pXO1aat which are variable for this sample of B. anthracis cultures showed that all strains divided on 2 main clusters - A and B. Cluster A consisted of 5 genotypes whereas cluster B - of 1 genotype. All highly virulent original strains and variants with characteristic phenotype Cap(CO2)(+)(O2)(-)Tox(+)ProtA(+)Hly(+) Lec(-)Trp(+) had identical genotype in 4 groups and in 5th group differences were present only in vrrA locus. All original strains and variants with the most atypical complex of phenotypic characteristics Cap (CO2)(+)(O2)(+)Tox(-)ProtA(-)Hly(-)Lec(-)Trp(-) also had the same genotype belonging to cluster B and diverged on characteristic of 5 chromosomal VNTR locuses and pXO1aat locus from typical strains. Absence of toxin production in vitro was not related to loss of genetic determinants of toxin components. Cultures with typical characteristics, one of which was ability to produce toxin in vitro, had larger sizes of amplicons of pXO1aat locus (135 and 132 nbp), whereas atoxigenic original strains and variants with complex of atypical characteristics and identical chromosome genotype had the smallest sizes (123 bnp). All original cultures were isolated in Russia, their genotypes are described for the first time.

[Properties of live antibiotics-resistant anthrax vaccine STI-PR after long-term storage].

Study showed that cultural, morphologic, genetic, immunologic characteristics, and resistance to antibiotics of STI-PR anthrax vaccine did not change after storage during 20 years in lyophilized condition. It has been shown that medium for lyophilization plays important role in preservation of vitality of anthrax spores. Optimal preservative properties have been observed for thioureal and sucrose-gelatinous media for lyophilization. Obtained results give reasons for prolongation of shelf live of STI-PR vaccine from 2 - 3 to 5 - 8 years.

[Characterization of Bacillus anthracis typical virulent test strain 81/1].

The results of the prolonged and many-sided study of B. anthracis strain 81/1 by different authors are presented. The cultural and morphological, biochemical, antigenic, molecular-genetic characteristics of this strain give grounds for regarding it as a typical test strain to be used for the determination of the vaccines immunogenicity, the effectiveness of antibiotics and immunomodulators.

[Epidemiological situation on anthrax in the regions of the Southern Federal District in connection with the flood in June 2002]. / Epidemiologicheskaia obstanovka po Sibirskoi iazve v regionakh Iuzhnogo Federal'nogo Okruga v sviazi s navodeniem v iiune 2002 g.

Data on the epidemiological situation in anthrax in the regions of the Southern Federal District, situated in the zone of inundation, are presented. In 2002 no epidemic complications in anthrax were noted in the affected territories. The causes contributing to the appearance of anthrax cases among humans and animals are presented. Recommendations on the improvement of anti-epidemic measures are given.

[The nonspecific resistance indices of different species of laboratory animals immunized with STI anthrax vaccine]. / Pokazateli nespetsificheskoi rezistentnosti u razlichnykh vidov laboratornykh zhivotnykh, immunizirovannykh sibireiazvennoi vaktsinoi STI.

A complex comparative study of the characteristics of nonspecific resistance in different species of laboratory animals immunized with vaccine STI against anthrax has revealed the existence of marked interspecific differences between noninbred white mice, guinea pigs, rabbits and noninbred white rats in such characteristics as phagocytic activity, oxygen-dependent function of polymorphonuclear blood leukocytes, serum beta-lysin and lysozyme.

[Comparative multifactorial analysis of combined administration of injection and peroral forms of an antibiotic with a microbial immunomodulator in experimental anthrax]. / Sravnitel'nyi mnogofaktornyi analiz kombinirovannogo primeneniia in''ektsionnoi i peroral'noi lekarstvennykh form antibiotika s immunomoduliatorom pri éksperimental'noi sibireiazvennoi infektsii.

Comparative efficacy of the use of injection and oral dosage forms of rifampicin in the subtherapeutic doses in combination with peptidoglycan , an immunomodulator of microbial origin, was studied in respect to experimental anthracic infection with application of multifactorial analysis. It was shown that the antibiotic and immunomodulator had a pronounced synergistic effect. Polynomial statistic models were developed and nomograms or equal level curves defining the survival rate and average life-span (ALS) of the experimental animals within a wide range of the antibiotic and immunomodulator doses and the peptidoglycan dosing time were plotted. The combined use of the injection rifampicin in the subtherapeutic doses and the immunomodulator provided a significant increase in the survival rate and ALS, whereas the use of the oral antibiotic in combination with the immunomodulator increased only the ALS and not the survival rate. Multifactorial analysis proved to be an optimal methodical approach to comparative study of various antibiotic dosage forms used in combination with immunomodulators under experimental conditions.

[Multifactorial experiment on the combined action of doxycycline and a low molecular weight immunomodulator of microbial origin in experimental anthrax infection]. / Izuchenie v mnogofaktornom éksperimente sochetannogo deistviia doksitsiklina i nizkomolekuliarnogo immunomoduliatora mikrobnogo proiskhozhdeniia pri éksperimental'noi sibireiazvennoi infektsii.

Chemotherapeutic efficacy of combined therapy of experimental anthrax infection with subtherapeutic doses of doxycycline and a low molecular weight immunomodulator of microbial origin was studied with mathematical design of the experiment and multifactorial analysis. A marked synergistic effect of oral doxycycline and the immunomodulator was observed. The results of the multifactorial experiment were computer processed and polynomial statistic models (the second order equations) describing the survival rate and mean lifespan (MLS) were derived. The equal level lines characterizing the survival rate and MLS were plotted against the fixed values of the time factor of administering the immunomodulator and the dose of the antibiotic. The doses of the immunomodulator and the time of its administration were optimized with respect to the maximum therapeutic effect with doxycycline subtherapeutic doses.

[The assessment of antitoxic anti-anthrax immunity]. / Otsenka antitoksicheskogo protivosibireiazvennogo immuniteta.

In experiments on guinea pigs immunized with avirulent noncapsular strains STI, Sterne (34F2) and the avirulent mutant of Bacillus anthracis strain 228/8 the relationship between the titers of serum antibodies to the preparations of purified protective antigens (PA) and purified lethal factor (LF) of B. anthracis toxin and the level of the antitoxic activity (ATA) of blood sera, as well as acquired resistance, was analyzed. The ATA of sera was evaluated in the primary culture of peritoneal macrophages affected by the mixture of PA and LF. The level of relationship (r) between individual ATA values and the titers of antibodies to PA and LF was shown to vary over a wide range, depending on the group of the animals and did not exceed, on the average, 0.19-0.37. At the same time the mean values of these characteristics, followed in their dynamics depending on the immunogenic properties of vaccine strains or the time elapsed after vaccination, were highly correlated (r = 0.76-0.87). The possibility of using these characteristics for the evaluation of acquired resistance are discussed.

[The effect of the protective antigen of Bacillus anthracis on the formation of immunity under the action of live anthrax vaccines]. / Vliianie protektivnogo antigena Bacillus anthracis na formirovanie immuniteta pod deistviem sibireiazvennykh zhivykh vaktsin.

The preparation of low-molecular protective antigen (PA) isolated from strain 34F2 (Sterne) and having a molecular weight of 34 and 51 kD, unlike the preparation of high-molecular PA with a molecular weight of 87 kD, suppressed the formation of acquired resistance to anthrax when introduced into guinea pigs in mixture with live spores of strains of STI, 34F2 and new vaccine strain 228/8; this phenomenon was mainly accompanied by a decrease in the level of antibodies to lethal factor (LF) nad in the antitoxic activity of blood serum. The immunosuppressing action of low-molecular PA depended on the kind of vaccine strain introduced together with this antigen, which suggested the existence of differences in the ligand determinants of strains 34F2 and STI. In contrast to high-molecular PA, low-molecular PA blocked the action of the lethal mixture of PA and LF on the culture of peritoneal mononuclear phagocytes of CBA mice. The competitive relationships between low-molecular PA and high-molecular PA are discussed.

[Methodological characteristics of the use of the immunofluorescence method for the rapid determination of the sensitivity of the anthrax microbe to antibiotics]. / Metodicheskie osobennosti ispol'zovaniia immunofliuorestsentnogo metoda dlia ékspressnogo opredeleniia chuvstvitel'nosti sibireiazvennogo mikroba k antibiotikam.

An immunofluorescent method for rapid assay of antibiotic sensitivity of Bacillus anthracis was tested with the use of virulent strains. It was shown that the immunofluorescent method was applicable for assay of antibiotic sensitivity of Bacillus anthracis immediately upon inoculation of the native matter: soil samples and other materials. Comparison of the results obtained with the method of serial dilutions and the immunofluorescent method showed that the levels of the Bacillus anthracis sensitivity to rifampicin, benzylpenicillin, tetracycline, streptomycin and gentamicin coincided. The immunofluorescent method provided the results on antibiotic sensitivity of Bacillus anthracis 6-8 hours after initiation of the rapid assay, the inoculum size being at least 10(6) spores/ml. Isolation of the causative agent pure cultures was not required. Under laboratory conditions the assay required consideration of the characteristic features of the causative agent and fixation of smear replicates in a mixture of 96 degrees ethyl alcohol and 3 per cent hydrogen peroxide for 30 min. After the assay the panels should be disinfected by immersing for 16-18 hours into 6 per cent hydrogen peroxide supplemented with 0.5 per cent of a detergent.