[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.

A NEW GENETIC VARIANT OF THE CRIMEAN-CONGO HEMORRHAGIC FEVER VIRUS ISOLATED IN CRIMEA.

This work represents the results of the genetic identification of the Crimean-Congo hemorrhagic fever virus (CCHF virus) strains isolated in the Crimean Federal District in conducting the epidemiological survey of the imported case of CCHF from Crimea in 2015. One sample of the serum from a patient and 61 pools (506 specimens) of ticks collected during the epizootiological survey of 6 administrative districts of the Crimean Federal District were tested using PCR for the presence of the CCHF virus RNA. RNA of the CCHF virus was detected in serum from a patient and 10 samples of ticks. The genetic identification of the CCHF virus was performed by sequencing the virus genome S-, M-, and L-segments. The result of the molecular-genetic analysis revealed a high degree of identity between the samples of the CCHF virus in human serum and three samples of ticks and their belonging to a new genetic Crimea subclade (Vd) of the genotype Europe 1. Whole genome sequencing of two samples of CCHF virus belonging to the Crimea subgroup (Vd) was performed. CCHF virus variants of the Crimea subclade (Vd) of the Europe- lgenotype were described for the first time. These variants were endemic to the territory of the Crimean peninsula.

[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.

[Improvement of methods for identification of atypical anthrax strains and their differentiation from closely related bacilli].

AIM: To study biologic characteristics of atypical strains of anthrax agent in order to improve methods of identification and differentiation from closely related bacilli. MATERIALS AND METHODS: Objects of the study were 1101 strains of microorganisms of which following were experimentally examined: atypical--40 strains, typical--2, saprophytic bacilli from Bacillus genus--50. Aside from conventional methods, additional tests for intraspecies differentiation as well as multiplex PCR method were used for identification. RESULTS: Isolation rate of atypical strains of anthrax agent in natural conditions as well as frequency of misidentification of bacillary strains as Bacillus anthracis was assessed. Phenotypical test for determination of susceptibility to penicillin was improved. Variant of multiplex PCR for differentiation of B. anthracis strains with any set of plasmids from closely related bacilli strains was developed. Feasibility to use multiple loci analysis of 6 chromosomal and 2 plasmid regions of B. anthracis genome containing variable number of tandem repeats (MLVA) for differentiation of B. anthracis strains from other bacilli from Bacillus genus was demonstrated. CONCLUSION: In order to optimize the processes of identification of B. anthracis typical and atypical strains and differentiation between closely related bacilli, it is rational to use disk-diffusion method with commercially available disks of penicillin, multiprimer PCR and MLVA on 6 chromosomal and 2 plasmid loci.

[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.

[Germination of Bacillus anthracis spores].

The influence of amino acids, nucleosides and inorganic components on the kinetics and effectiveness of the germination of B. anthracis spores was studied. The study revealed that the rapid germination of the spores took place after their activation at 65 degrees C in tris buffer with L-alanine in combination with inosine or adenosine added; less pronounced germinative action was caused by the addition of alanine only and the combination of phenylalanine, tyrosine and tryptophan. The rapidity of germination and the sets of effective germinants for spores of different strains were different. All B. anthracis strains under study had nucleotide sequences, of gene gerX in their genome.

[Multiplex amplification test system for the identification and differentiation of Bacillus anthracis].

The multiplex amplification test system for the identification of Bacillus anthracis with primers to plasmid cya (pX01), capC (pX02) genes and chromosomal sap gene were developed. The primers to sap gene were selected by the authors and, after being tested on 72 microbial strains of the genus Bacillus, proposed as more specific in comparison with the known primers to chromosomal locus Ba 813. The proposed test system permitted the simultaneous identification of B. anthracis of all plasmid variants, the evaluation of their potential virulence and the differentiation of B. anthracis nonplasmid strains from bacilli of the group Bacillus cereus.

[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.

[Gene typing of Bacillus anthracis strains isolated on the territory of the countries of the Confederation of Independent States]. / Genotipirovanie shtammov Sibireiazzvennogo mikroba, izolirovannykh na territorii stran SNG.

Thirty eight B. anthracis strains isolated on the territory of the former USSR from different sources at different periods were studied by the method of the multilocus analysis of 6 chromosomal and 2 plasmid regions of B. anthracis genome with a variable number of tandem repeats. The strains belonged to 18 different genotypes; of these, 14 genotypes were described for the first time. The analysis of the genetic relationship of the strains gave grounds to suggest that on this territory both closely related strains and strains whose genotypes were remote from those peculiar to the greater part of other strains could occur. The strains belonging to subgroup A1a of molecular variability were "endemic" for the European part of the former USSR. A modification of the method of gene typing was proposed, which permitted it to be made without the use of an automatic sequencer; this made it possible to greatly widen the circle of laboratories where this method of research could be used.

[Clinical experience in the use of clonidine in regional anesthesia and early postoperative period (1984-1989)]. / Klinicheskii opyt primeneniia klonidina v regionarnoi anestezii i rannem posleoperatsionnom periode (1984-1989 gg.).

Clonidine attracts attention as a sole agent for local anesthesia. This review analyzes the role of alpha 2-adrenergic agents in modern regional anesthesiology, both practical and experimental. Recent controlled studies showed that an alpha 2-adrenoreceptor clonidine administered spinally is characterized by a dose-dependent antinociceptive effect and is as effective as epinephrine in prolonging local anesthetic blocks. Moreover, in combination with spinal opioids, intraspinal clonidine possesses a synergic analgesic effect, potentiating the intensity and duration of opioid anesthesia. Use of clonidine as an adjuvant to various plexus blockades and for caudal block in children is discussed. Regional administration of clonidine involves side effects, such as hypotension, bradycardia, and sedation. Minimization of the dose of alpha 2-adrenergic agonists minimizes the complications. Regional clonidine effects (analgesia, hemodynamic changes, and sedation) and their recognized mechanisms of action are described.