[Experimental non-culturable Vibrio cholerae eltor and its biological properties]. / Eksperimental'noe poluchenie nekul'tiviruemykh form Vibrio cholerae eltor i kharakteristika ikh biologicheskikh svoistv.

In experiments with the cultivation of V. cholerae eltor under the conditions of high salt concentration, as well as low temperature and deficiency in nutrient substances, uncultivable forms (UF) of toxigenic and nontoxigenic vibrios were obtained. The absence of growth of seeded vibrios after the filtration of samples (with a filter of 0.22 micron), the preservation of specific antigenic determinants and the initial set of genes, changes in the morphology of cells (small size, coccoid form with the flagella retained) confirm the transition of V. cholerae eltor under study into the uncultivable state which, under unfavorable conditions, more rapidly develops in toxigenic vibrios than in nontoxigenic ones. The analysis of the INT-reductase activity of UF disintegrates revealed that they had endogenic respiration whose activity increased (4.5- to 6.5-fold) in the presence of the exogenic intermediates of the Krebs cycle. The uncultivable forms of the vibrios retain genes responsible for pathogenicity, as well as their antigenic determinants.

[Dot immunoassay with colloidal silver particles as a marker of specific antigen for testing human sera for brucellosis]. / Dot-immunoanaliz s chastitsami kolloidnogo serebra v kachestve markera spetsificheskogo antigena dlia issledovaniia syvorotok krovi liudei na brutsellez.

Whether the dot immunoassay is suitable for the detection of Brucella antibodies in human sera by using a colloidal silver-labeled Brucella specific antigen as a diagnostic tool is assessed. The antigen was the B. abortus 19BA protein polysaccharide complex isolated by Brucella acetic acid hydrolysis. The dot immunoassay is easy-to-use, cost-effective, highly sensitive, and therefore of more informative value in detecting Brucella antigens than routine serological tests (Huddleson test, Wright agglutination test, passive hemagglutination test, long-term complement fixation test, and Coombs test). It requires no use of expensive equipment and reagents.

[Use of specific antibodies labelled with colloid silver particles for detection of Brucella antigens with dot immune analysis]. / Ispol'zovanie spetsificheskikh antitel, mechennykh chastitsami kolloidnogo serebra, dlia vyiavleniia antigenov brutsell metodom dot-immunoanaliza.

Fitness of dot immuno-analysis for detection of Brucella antigens labeled with colloid silver is evaluated. Soluble lipopolysaccharides and protein-saccharide antigen and corpuscular antigens of 22 Brucella strains (7 species) pathogenic for humans and animals in the S and R forms were used. The specificity of the method was tested on 10 heterologous microorganisms whose antigens were closely related. The suggested test system is simple, economic, highly sensitive (from 62 thousands to 8 million CFU/ml) and specific, requires no expensive equipment, and is an alternative to enzyme immuno-assay and dot immuno-analysis with gold immunosole.

[Comparative study of immunomodulating effects of natural experimental preparations on phagocytosis of Yersinia pseudotuberculosis]. / Sravnitel'noe izuchenie immunokorregiruiushchikh svoistv éksperimental'nykh preparatov prirodnogo proiskhozhdeniia pri fagotsitoze Yersinia pseudotuberculosis.

To correct defects of the body's phagocytic system, the authors used experimental agents, such as vital activity products of plant cells, which were obtained by biotechnology. These included arabinogalactan, syringin, K-212, and DFEGP. The study agents activate the oxidative metabolism of macrophages, the production of active forms of oxygen, and thus enhance its bactericidal effect against Yersinia pseudotuberculosis. Syringin and arabinogalactan produce a more pronounced stimulating macrophageal effect in Yersinia pseudotuberculosis phagocytosis than do K-212 and DFEGP. The findings indicate that further studies of the effects of these tested and other agents on humoral and cellular immunity are required.

[Detection of brucella antigens with colloid metal particles as markers for specific antigens]. / Obnaruzhenie antigenov brutsell s pomoshch'iu chastits kolloidnykh metallov v kachestve markerov spetsificheskikh antitel.

Gold and silver sols were comparatively approved as markers of specific IgG isolated from hyperimmune Brucella antisera for the detection of brucellar antigens. The sensitivity of the test system using gold immunosol proved to be some higher (3.1-9.8 ng/ml of soluble and 2.0 x 10(4)-5.3 x 10(6) CFU/ml of corpuscular brucellar antigens) than that achieved with the use of silver immunosol (5.7-18.4 ng/ml of soluble and 6.1 x 10(4)-8.0 x 10(6) CFU/ml of corpuscular brucellar antigens). At the same time silver sol was a cheaper and more available marker. Both test systems were found to be highly specific. False positive results were observed only with Yersinia enterocolitica O:9 at high concentrations due to the fact that they had common polysaccharide haptens incorporated into lipopolysaccharides of these microorganisms. The proposed test systems with colloid metal particles used as markers of specific antibodies for the detection of Brucella antigens are technologically simple, economic, rapid, highly sensitive and specific. Their use in combination with other serological methods will make the results of analyses more informative, thus improving the quality of laboratory diagnostics of brucellosis.

[Cholera in the Far East of Russia. Communication 1. Epidemiological characteristics of the outbreak of cholera El tor in Vladivostok]. / Kholera na Dal'nem Vostoke Rossii. Soopshchenie 1. Epidemiologicheskaia kharakteristika vspyshki kholery él'tor v g. Vladivostoke.

The materials on the investigation of the outbreak cholera eltor in Vladivostok, caused by the import of infection from China. The leading role of the water route of transmission of this infection is shown due to the contamination of water sources with non-decontaminated sewage water. The complex of antiepidemic measures was carried out, which made it possible to arrest the spread of cholera and liquidate its foci.

[Analysis of PCR-diagnosis of brucellosis in human]. / Osobennosti PTsR-diagnostiki brutselleznoi infektsii u liudei.

Polymerase chain reaction (PCR) was used for the diagnosis of brucellosis in humans with different forms of this disease. A high incidence (77.6%) of Brucella infection was revealed in the staff of cattle breeding centers with unfavorable situation with regard to brucellosis. Such a conclusion was made after PCR testing of native human sera. In acute brucellosis of humans amplification of the specific site of brucella DNA in PCR is possible only after extraction of DNA by a procedure adapted for DNA extraction from intact brucella cells. In chronic infection weak amplification of brucella genome DNA fragment was observed in investigation of native sera by the PCR. More expressed amplification product was recorded in PCR with a DNA precipitate from this serum obtained by ethanol precipitation. A still higher level of brucella DNA fragment amplification was observed after DNA extraction from sediment obtained by ethanol precipitation from this serum. These data confirmed the incomplete phagocytosis phenomenon at the early stage of infection, known in brucellosis pathogenesis, and allowed some hypotheses on the pathogenesis of chronic phase of brucellosis infection.

[Detection of Brucella antigens by dot immunoassay using specific antibodies labelled with colloid gold particles]. / Vyiavlenie antigenov Brutsell metodom dot-immunoanaliza s ispol'zovaniemspetsificheskikh antitel, mechennykh kolloidnym zolotom.

Dot immunoassay was developed to improve the quality of laboratory diagnosis of brucellosis. Particles of colloid gold were used as a marker of specific antibodies. The method was used for detecting Brucella antigens in artificially contaminated environmental objects (soil and water) and in biological material (milk, blood serum, and visceral homogenates of animals). The sensitivity of the test system was 19.5.10(3)-62.5.10(4) CFU/ml. Specificity of the assay was tested with 10 heterologous antigenically closely related bacterial species. The proposed test system is simple, economic, highly sensitive and specific, and requires no expensive equipment and reagents.

[Use of protein-polysaccharide Brucella antigen labeled with colloidal gold for detection of specific antibodies by the dot-immunoassay method]. / Ispol'zovanie belkovo-polisakharidnogo antigena brutsell, mechennogo chastitsami kolloidnogo zolota, dlia obnaruzheniia spetsificheskikh antitel metodom dot-immunoanaliza.

Antibrucella antibodies were detected by dot-immunoassay with colloid gold label of antigen. The specific antigen was protein-polysaccharide complex (PPC) isolated from vaccine strain Brucella abortus 19 BA by acetic acid hydrolysis of bacterial cells. Dot immunoassay with PPC labeled with colloid gold was effective in testing cattle, rabbit, and human sera for antibrucella antibodies. The proposed test system is simple, economic, highly sensitive and specific, and requires no expensive equipment and reagents, which are its advantages over routine serological tests.

[Comparative study of various methods for determining Vibrio cholerae toxigenicity]. / Sravnitel'noe izuchenie razlichnykh metodov opredeleniia toksigennosti Vibrio cholerae.

Testing of 138 Vibrio cholerae strains for gene determinants responsible for the production of cholera enterotoxin by the polymerase chain reaction (PCR) and gene probing using molecular CT-probe showed good correlation of the results of different methods and correlation of these data with studies of V. cholerae strain virulence in vivo and in hemolytic activity test. The advantages of PCR in rapid assessment of the toxigenicity and epidemic significance of V. cholerae strains are demonstrated.

[Development of a method for alkaline extraction of DNA from Brucella for diagnosing brucellosis using the polymerase chain reaction]. / Razrabotka metodov shchelochnoi ékstraktsii khromosomnoi DNK iz Brutsell dlia diagnostiki brutselleza s pomoshch'iu metoda polimeraznoi tsepnoi reaktsii.

Several methods of alkaline extraction of chromosome DNA from Brucella in the presence of 50 microliters model diagnostic material blood serum are developed for the diagnosis of brucellosis by the polymerase chain reaction (PCR). These methods are based on the capacity of NaOH to effectively denature proteins and destroy Brucella cell wall, thus isolating the genome DNA without exposure to proteolytic enzymes, detergents, deproteinization, or pH neutralization. The first method consists in alkaline lysis by 0.2-1.0 M NaOH followed by DNA precipitation with two ethanol volumes in the presence of 0.1 M NaCl, washing of the resultant precipitate in 80% ethanol, drying of the precipitate, and dissolving in distilled water. The second method includes alkaline lysis in the presence of 0.3 M NaCl with NaOH concentrations of 0.5-4.3 M and the stages of DNA sedimentation, washing of precipitate, its drying and dissolving similar to those in alkaline lysis. The third method consists in alkaline lysis-precipitation by 0.2-05 M NaOH in the presence of 0.1 M NaCl and 64% ethanol, followed by DNA preparation stages similar to those in alkaline lysis. The best results were achieved by alkaline lysis in the presence of 0.3 M NaCl at NaOH concentrations of 0.7 and 2.1 M, which meant theoretical levels of sensitivity 140 and 86 Brucella cells, respectively.

[Characteristics of phagocytosis of Yersinia pseudotuberculosis with various sets of plasmids]. / Osobennosti fagotsitoza Yersinia pseudotuberculosis s raznym naborom plazmid.

The study used the selected aplasmid strain of Y. Pseudotuberculosis 53 and I-716, which contains plasmids that have molecular weights of 48 and 82 mD, and I-727 that has only virulence plasmid pYV48. The study has indicated that the fact that Yersinias have pVM82 and pYV48 enhances their resistance to phagocytosis. The Y. pseudotuberculosis strains carrying these plasmids suppress the responsiveness of phagocytes, inhibit an "oxidative explosion", lower the activities of superoxide dismutase and myeloperoxidase, thus suppressing their bactericidal capacity.

[Use of specific antibodies, labeled with colloidal gold particles, for the detection of Brucella antigens using dot-immunoassay]. / Ispol'zovanie spetsificheskikh antitel, mechennykh chastitsami kolloidnogo zolota, dlia obnaruzheniia antigenov brutsell metodom dot-immunoanaliza.

The method of dot immunoassay with the use specific antigens, labeled with colloidal gold particles, for the detection of brucellar antigens was developed and tested under laboratory and field conditions. In this work soluble antigens isolated from different Brucella species and corpuscular antigens (13 strains belonging to 7 species of the genus Brucella, most pathogenic for humans and animals in the S- and R-forms) were used. The method was tested in the study of pathological material obtained from sick animals and humans in a farm with unfavorable situation for brucellosis in the Irkutsk Region. The sensitivity of the proposed assay system was found to be high and constituted 10 pg/ml to 1 ng/ml for soluble brucellar antigens and 200 CFU/ml to 13.5 x 10(6) CFU/ml for corpuscular antigens of Brucella S- and R-forms. The specificity of the method was tested with the use of 10 heterologous microorganisms. False positive results were observed only with Yersinia enterocolitica 0:9 at a concentration of 1 x 10(6) CFU/ml due to similarity of their polysaccharide-containing surface antigens. The newly developed dot immunoassay is simple for use, rapid and does not require expensive reagents and equipment.

[The characteristics of the protectiveness of a preparation of Vibrio cholerae outer membrane based on the data from a study using the ligated intestinal loop]. / Kharakteristika protektivnosti preparata naruzhnoi membrany kholernogo vibriona po dannym issledovaniia pereviazannoi kishechnoi petli.

The study aimed at finding out the antiadhesive capacity of antigenic preparation, earlier obtained from V. cholerae outer membrane and highly effective with respect to cholera infection, was undertaken. The study was made on previously immunized adult rabbits who had been subjected to laparotomy under anesthesia and the ligation of intestinal loops, subsequently inoculated with the broth culture of V. cholerae eltor (P-3122, serovar Ogawa). The intestinal loops were studied histologically and bacteriologically with the calculation of the number of vibrios, the deduction of the adhesion index and the coefficient of the efficacy of immunization. The data thus obtained indicated that the specific immunization of rabbits with their subsequent inoculation with V. cholerae virulent strain suppressed the adhesive activity of the infective agent which was more pronounced in rabbits immunized with the preparation of V. cholerae outer membrane.

[Adaptive mechanisms in the ecology of the pseudotuberculosis microbe]. / Adaptivnye mekhanizmy v ékologii psevdotuberkuleznogo mikroba.

The paper presents the radiorespirometric and enzymatic findings of the insensitivity of catabolism of glucose and acetate in Y. pseudotuberculosis cells. The reduction in a Y. pseudotuberculosis cultivation temperature (4 degrees C) has been found to be followed by higher activities of NADP(+)-dependent dehydrogenases and increased functioning of the glyoxylate cycle. The temperature-compensated increase in the functioning intensity of alternative metabolic pathways appears to be part of adaptative metabolic manifestations in the Y. pseudotuberculosis cells during autonomic habitance and accumulation in the environmental objects.

[The outer membranes of Vibrio cholerae as a potential component in a chemical vaccine]. / Naruzhnye membrany kholernogo vibriona kak potentsial'nyi komponent khimicheskoi vaktsiny.

The results of the study of the preparation of V. cholerae eltor membrane, obtained by the lysis and inactivation of microbial cells with urea and the subsequent differential centrifugation and nuclease treatment. As revealed in this study, the outer membrane preparation, when introduced parenterally and orally to mice, induced pronounced immunity to experimental cholera infection and the production of vibriocidal antibodies in high titers. The treatment of V. cholerae eltor membranes with trypsin led to further increase of the immunogenic potency of the preparation. The protective action of V. cholerae eltor outer membranes considerably exceeded the protective effect of currently used whole-cell eltor vaccine. This opens prospects for using the above-mentioned preparation for the improvement of chemical vaccine as a component ensuring the formation of antibacterial immunity.

[The activity of the phagocytic bactericidal systems in intact guinea pigs and in guinea pigs immunized against tularemia]. / Aktivnost' bakteritsidnykh sistem fagotsitov u intaktnykh i immunizirovannykh protiv tuliaremii morskikh svinok.

The activities of NADP-oxidase and glucose-6-phosphate dehydrogenase in mononuclear phagocytes and polymorphonuclear leukocytes (PML) during the phagocytosis of Francisella tularensis and the influence of immunization of on the activity of oxygen-dependent metabolism were studied. The phagocytosis of F.tularensis was found to be accompanied by a rise in the activity of NADP.H-oxidase and glucose-6-phosphate dehydrogenase. In the process of immunogenesis the rearrangement of the enzymatic system ensuring the antimicrobial action of phagocytes occurred. Macrophages exhibited higher enzymatic activity than PML.

[Electron microscopic data on the action of Yersinia pestis bacterial toxin on phagocytosis]. / Elektronno-mikroskopicheskie dannye o deistvii toksina bakterii Yersinia pestis na fagotsitoz.

Electron-microscopic study of the interaction of Y. pestis and peritoneal phagocytes obtained from BALB mice, both intact and treated with the toxic fraction of Y. pestis, has revealed that in the process of phagocytosis the toxin weakens the killing and degradation phases of these bacteria, but leaves the ingestion activity of phagocytes unchanged. The process of phagocytosis in mice exposed to toxin is accompanied by destructive changes of cells and redistribution of myeloperoxidase, a bactericidal enzyme, in polymorphonuclear leukocytes without any signs of bacterial degradation.