[Corynebacterium diphtheriae nontoxigenic strain carrying the gene of diphtheria toxin]. / Kharakteristika netoksigennykh shtammov Corynebacterium diphtheriae, nesushchikh gen difteriinogo toksina.

Among 828 C. diphtheriae nontoxigenic cultures isolated in different region of Russia in 1994-2002, 114 cultures (13.8%) had the gene of diphtheria toxin (gene tox) and were thus called nontoxigenic tox-carrying (NTTC) strains. All NTTC strains were found to belong to biovar mitis and formed neither normal, nor "defective" diphtheria toxin. The most of NTTC strains (94%) belonged to ribotype "Moskva", not occurring among C. diphtheriae toxigenic strains. The incapacity of NNTC strains of forming diphtheria toxin was caused by mutation: the deletion of one nucleotide which led to the shift of the open reading frame and to the formation of the stop codon. The results of these studies are indicative of the fact that a sufficiently homogeneous and isolated group of C. diphtheriae nontoxigenic strains is spread in Russia. These strains carry the nonexpressing gene of diphtheria toxin and are of no epidemic importance in diphtheria infection.

[Genetic structure of Corynebacterium diphtheriae strains isolated in Russia during epidemics of various intensity]. / Geneticheskaia struktura shtammov Corynebacterium diphtheriae, vydelennykh v Rossii pri raznoi intensivnosti épidemicheskogo protsessa difterii.

The genetic structure of C. dipthteriae toxigenic strains isolated in Russia during the period of more than 50 years was analysed. The use of the method of ribotyping made it possible to register 17 C. diphtheriae ribotypes. The study revealed that the genetic structure of C. diphtheriae population varied in the dynamics of the epidemic process: each epidemic cycle characterized by predominant spread of epidemic strains of definite biovars and ribotypes. Thus, C. diphtheriae strains of biovar gravis, ribotype M11, dominated in the 40-60 years and C. diphtheriae strains of biovar mitis, closely related ribotypes M1 and M1v, dominated in the 80 years. During the last epidemic rise of diphtheriae morbidity in the 90 s C. diphtheriae strains of biovar gravis, closely related ribotypes G1 and G4, dominated among circulating strains. The proportion of these ribotypes began to increase 3 years before the rise of morbidity. The data of microbiological monitoring are recommended for use in the prognostication of the development of the epidemic process of diphtheria infection.

[Temperature-dependent changes in immunochemical properties of lipopolysaccharides of yersinia pestis]. / Temperaturozavisimye izmeneniia immunokhimicheskikh svoistv lipopolisakharida yersinia pestis.

The preparations of pilopolysaccharides (LPS) of virulent and avirulent Y. pestis strains, which have a different composition, were cultivated at different temperatures. Despite the cultivation temperature of parental cells, all LPS preparation inhibited the passive hemagglutination reaction (PHR) of the red blood cells sensitized by LPS isolated from the cultures grown at 26 degrees C by using homologous antisera. In contrast, the homologous system consisting of the red blood cells sensitized by LPS from the EV NIIEG, cultured at 37 degrees C, and the antiserum to these cells proved to be more specific and it was inhibited only by the homologous LPS isolated from the strain EV NIIEG. The similar reaction of the interaction of the red blood cells sensitized by high temperature LPS agents from plasmid-free strains with the same serum was inhibited by all plague LPS preparations. The LPS preparations from the strains Y. pestis 1146 and Y. pseudotuberculosis 9532 obtained when cultivated at 37 degrees C. RHR of the red blood cells sensitized by these preparations was inhibited by homologous LPS irrespective of the temperature of cell cultivation. In all cases, the reaction was specific for Yersinia strains and it was inhibited by LPS from Ra-Rd2 cultures, variants of Salmonella and E. coli.

[Comparative characterization of the physico-chemical properties of lipopolysaccharides of Yersinia pestis and R-mutants of enterobacteria]. / Sravnitel'naia kharakteristika fiziko-khimicheskikh svoistv lipopolisakharidov Yersinia pestis i R-mutantov énterobakterii.

Electrokinetic potentials (EKP) of the cells of R mutants of Escherichia coli and Salmonella minnesota and cells of Yersinia pestis strains EV (line NIIEG), 358/12 P-, TWJ, Java, and 231 (708) were determined, as well as EKP of lipopolysaccharide (LPS) preparations isolated from these bacteria. The electric characteristics of the cell surfaces of the strains under investigation were demonstrated to correlate with the LPS charge and the reduction extent of their molecules. Acidic hydrolysis of LPS on the cell surface resulted in the leveling of the distinctions in EKP values (their reduction to the same level). EKP values and the size of LPS micelles of the studied Y. pestis strains corresponded to those of the deep R mutants of enterobacteria, while the aggregation extent of the molecules was higher for Y. pestis.

[In vitro development of fluoroquinolone resistance in the glanders pathogen]. / Formirovanie vozbuditelem sapa ustoichivosti k ftorkhinolonam in vitro.

To improve the scheme for the therapy of malleus, the possible development of fluoroquinolone resistance in the cells of Pseudomonas mallei was studied. The frequency of the mutations determining the resistance to oxolinic acid, norfloxacin, enoxacin, ofloxacin and ciprofloxacin was detected. The spectrum of the cross resistance in the strains was investigated. The influence of the resistance mutations on the culture virulence was studied. Recommendations on the rational use of fluoroquinolones in the treatment of malleus are presented.

[Cloning and detailed mapping of the fra-ymt region of the Yersinia pestis pFra plasmid]. / Klonirovanie i detal'noe kartirovanie fra-ymt-oblasti plazmidy pFra Yersinia pestis.

The genetical libraries of the pFra plasmid of Yersinia pestis genes were obtained by insertion into the PstI, SalGI, EcoRI, XhoI restriction sites of the cosmid vector pHC79. The immunochemical analysis of the recombinant clones has revealed the clones synthesizing the antigen Fl (fraction I) and mouse toxin (Ymt--Yersinia pestis murine toxin). The restriction analysis of the plasmids from antigen synthesizing clones has permitted to construct the detailed physical map of the fra-ymt region of the pFra plasmid the size of 22 kb. The recombinant F1 positive clones of Escherichia coli are able to form at 37 degrees C the capsule-like structure peculiar for Yersinia pestis. The antigen F1 and the mouse toxin were isolated, purified and characterized. The antigen F1 is an 1-2 Md polymer containing a 16 kDa protein subunit. The mouse toxin a 240 kDa protein consisting of 61 kDa subunits. The nucleotide sequence of ymt gene has been defined.

[Genetic characteristics and protective properties of neamin-resistant mutant Salmonella typhimurium and S. dublin of the Nea(r) Str(s) and Nea(r) Str(r) 500 classes]. / Geneticheskaia kharakteristika i protektivnye svoistva neaminrezistentnykh mutantov Salmonella typhimurium i S. dublin klassov Nea(r) Str(s) i Nea(r) Str(r) 500.

The genetic analysis of attenuated mutants, class Nea(r) Str(s), with the use of bacteriophage P 22 has shown that mutation rendering the mutants resistant to neamine is localized in gene nea A. In experiments with the intraperitoneal infection of mice, the appearance of this mutation in S. typhimurium and S. dublin virulent strains has been found to lead to the decrease of virulence in 100% of clones. On the basis of the data obtained in this investigation, region str-spc in S. typhimurium and S. dublin has been mapped. In contrast to mutation spc A, mutations nea A and str A have been shown to inhibit the action of amber suppressor. The investigation has confirmed the regularity, previously established for Shigella flexneri, concerning the relationship between the influence of mutations, occurring in the genes which determine resistance to neamine and streptomycin and control the synthesis of ribosomal proteins S4, S5, S12 and S17, on the virulence of S. typhimurium and S. dublin and the effect of these mutations on the accuracy of the translation of genetic information in the biosynthesis of protein: mutation spc A has been found to produce no changes in the virulence of salmonellae, while mutations nea A and str A cause its loss. Salmonella strains carrying mutations nea A and nea B have shown pronounced protective properties in experiments on mice.

[Isolation and characteristics of the biological and genetic properties of neamin-resistant mutants of Salmonella typhimurium and Salmonella dublin candidates for vaccine creation]. / Poluchenie i kharakteristika biologicheskikh i geneticheskikh svoistv ustoichivykh k neaminu mutantov Salmonella typhimurium i Salmonella dublin, perspektivnykh dlia sozdaniia vaktsin.

Mutants, resistant to neamine and spectinomycin, have been isolated from S. typhimurium and S. dublin highly virulent strains. The neamine-resistant mutants can be divided into 3 classes in accordance with their sensitivity to streptomycin: sensitive, resistant to low and high concentrations of this antibiotic. The transduction analysis with the use of bacteriophage P 22 has revealed that the spectinomycin-resistant mutations under study are spc A mutations, while the mutations leading to resistance to neamine in class Near Strr 500 are nea B mutations. The mutation leading to resistance to spectinomycin (spc A) has been found to produce no changes in the virulence of salmonellae in the intraperitoneal infection of mice. The mutations leading to resistance to neamine and streptomycin (nea B and str A) have been found to decrease virulence.

[Auxotrophic mutations formed on the incorporation into the E. coli C600 chromosome of transposon Tnl that determines ampicillin resistance]. / Mutatsii auksotrofnosti, voznikaiushchie pri vstraivanii v khromosomu E. coli C600 transpozona Tnl, opredeliaiushchego ustoichivost' k ampitsillinu.

Incorporation of transposone Tnl of ampicillin resistance into the chromosome of E. coli C600 resulted in formation of at least 8 types of auxotrophic mutants. The transposone incorporates mainly into the loci the damage of which induces proline deficit (20 per cent). The sites of incorporation of transposone Tnl in the chromosomes of E. coli C600 and E. coli JC411 did not coincide.

[Isolation and characteristics of deletion mutants of thermosensitive plasmid pEG1]. / Vydelenie i kharakteristika deletsionnykh mutantov temperaturochuvstvitel'noi plazmidy pEG1.

Phenotypic revertants of temperature-sensitive in self-maintenance plasmid pEG1 derived from R-factor RP4 were studied. The study indicated a sertain part of them to be variants without ampicillin resistance. The frequency of pEG1 ampicillin sensitive (Aps) derivatives is about 5.10(-7) and does not depend on the recA gene product. The Aps-derivatives of pEG1 plasmid can be classified in two phenotypic groups. The first group is temperature-independent and does not inhibit the growth of host cells by non-permissive temperature (43 degrees C). Agarose gel electrophoresis has revealed that plasmids of this group occur as a result of deletions in plasmid pEG1. The other group of Aps-derivatives still possess the property to be eliminated at 43 degrees C and they inhibit the growth of bacterial cells like pEG1. Agarose gel has demonstrated that some of them are deletion mutants while others are quite similar to pEG1 plasmid. The data obtained in the course of study of deletion mutants make it possible to suppose that the presence of gene with temperature sensitive mutation in pEG1 is not necessary for stable maintenance of R-factor.

[Preparation and comparative evaluation of experimental anthrax diagnostic sera in experiments on animals]. / Poluchenie i sravnitel'naia otsenka éksperimental'nykh sibireiazvennykh diagnosticheskikh syvorotok v opytakh na razlichnykh zhivotnykh

The authors present the results of studies on obtaining and comparative assessment of experimental anthrax diagnostic sera in experiments on various animals. Donkeys, sheep, horses, rabbits and monkeys (Papio hamadryas) were immunized with the STI-I vaccine by a single scheme. The activity of the obtained sera was tested in the diffuse precipitation reaction by the amount of the detected antibodies and the titre. The most active sera were obtained from donkeys and sheep: their titre was 5.5 and 4 times greater and amount of the detected antibodies 2.6--2 times greater than in the sera of horses used up to the present time as producers of diagnostic anthrax preparations. The least active sera were obtained from monkeys.