RESUMEN
AIM: Evaluation of quality indicators of constructed cholera antigen polymer diagnosticums by using a complex of specific anti-cholera sera. MATERIALS AND METHODS. Cell lysates of cholera vibrio strains Vibrio cholerae cholerae 1395, V. eltor Ogawa 2044, V. eltor Inaba 13020, V. cholerae O139 16064 were sensitins for experimental preparations. 3 sera from cholera patients, normal human sera, cholera O1 (Ogawa, Inaba) commercial horse, cholera O139 commercial rabbit and heterologic sera against shigella, salmonella, escherichia and yersinia as well as experimental cholera rabbit sera against O1 and O139 were used as control. RESULTS: The study established that diagnosticums based on V. cholerae cholerae 1395 and V. cholerae O139 16064 strain sensitins by quality indicators may be used in the future for construction of these diagnosticums. CONCLUSION: Antibody containing preparations--commercial horse O1 sera, rabbit experimental and commercial sera and MCA O139 demonstrating titers not lower than 1/5120-1/10240 may serve as a control of experimental diagnosticums in the absence of human sera from cholera patients.
Asunto(s)
Antígenos Bacterianos/química , Cólera/diagnóstico , Inmunoensayo , Polímeros/química , Serotipificación/normas , Vibrio cholerae/química , Animales , Antígenos Bacterianos/inmunología , Cólera/sangre , Cólera/microbiología , Caballos , Humanos , Sueros Inmunes/química , Sueros Inmunes/inmunología , Conejos , Juego de Reactivos para Diagnóstico/normas , Serotipificación/métodos , Vibrio cholerae/inmunologíaRESUMEN
The in vitro study of the adhesive properties of V. cholerae eltor and V. cholerae O139 on a model of mammalian red blood cells revealed a correlation of their adhesive properties, the presence of the ctx AB, tcpA genes, and their hemolytic activity when blood group A (II) red blood cells were used. In the latter case, the strains having the characteristics of ctx(+) tcp(+) Hly(-) were ascertained to have a mean adhesive value (MAV) of > 1.5, a red blood cell involvement coefficient (RBCIC) of > 50%, while those with the characteristics of ctx(-) tcp(-) Hly(+) had a MAV of < 1.5 and a RBCIC of < 50%. In terms of the adhesive activity, the cultures with the genotype ctx tcp(+) Hly(+) is a heterogeneous group and may be low (MAV < 1.5) and high (MAV > 1.5) adhesive. According to the data of our studies, their adhesiveness is associated with the region of identification and independent on its object.
Asunto(s)
Eritrocitos/fisiología , Vibrio cholerae/fisiología , Animales , Adhesión Bacteriana , Técnicas Bacteriológicas , Cólera/epidemiología , Cólera/microbiología , Métodos Epidemiológicos , Eritrocitos/microbiología , Proteínas Fimbrias/genética , Humanos , Reacción en Cadena de la Polimerasa , Conejos , Ovinos , Especificidad de la Especie , Vibrio cholerae/clasificación , Vibrio cholerae/patogenicidadRESUMEN
Hemolysin of ctx+ Vibrio cholerae strains was obtained and studied. Ctx+ Vibrio cholerae O1 and O139 strains produced the hemolysin during cultivation in triptone medium without FeCl3. Mol.wt. and the spectrum of lytic activities of hemolysins of ctx+ Vibrio cholerae did not differ from hemolysins of ctx- strains.
Asunto(s)
Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Proteínas Hemolisinas/química , Proteínas Hemolisinas/metabolismo , Vibrio cholerae O139/metabolismo , Vibrio cholerae no O1/metabolismo , Animales , Proteínas Bacterianas/aislamiento & purificación , Caribdotoxina/genética , Toxina del Cólera/genética , Medios de Cultivo , Eritrocitos/metabolismo , Compuestos Férricos , Cobayas , Proteínas Hemolisinas/aislamiento & purificación , Hemólisis , Peso Molecular , Conejos , Ovinos , Vibrio cholerae O139/genética , Vibrio cholerae O139/crecimiento & desarrollo , Vibrio cholerae no O1/crecimiento & desarrolloRESUMEN
The influence of hydrogen peroxide on the dynamics of transition into uncultivable state (UCS) and on the reversion of V. cholerae and their subcultures, resistant to hydrogen peroxide, was studied. The transition of the initial cultures in river and distilled water into UCS took place earlier than that in resistant to hydrogen peroxide variants. The capacity for reversion to hydrogen peroxide resistant subcultures preserved, on the average, 2 - 3 times longer. An increase in the level of hydrogen peroxide in uncultivable populations was found to be 2.7 - 4.4 times. Subcultures, resistant to hydrogen peroxide, in the vegetative form had lower characteristics of peroxide concentrations than in uncultivable form (UCF), but somewhat higher than in initial variants. In revertants the concentration of hydrogen peroxide was lower in UCF, but somewhat higher than in vegetative cultures. The dynamics of the formation of UCF by cholera vibrios, with different degree of stability to the action of hydrogen peroxide, the accumulation of hydrogen peroxide in uncultivable populations, the deceleration of transition into uncultivable forms, an accumulation of hydrogen peroxide and an increase in the time of the reversion of clones, resistant to hydrogen peroxide, made it possible to suggest that the accumulation of hydrogen peroxide was possible to make an essential contribution to the formation of UCF of cholera vibrios in an experiment.
Asunto(s)
Peróxido de Hidrógeno/farmacología , Oxidantes/farmacología , Vibrio cholerae O1/efectos de los fármacos , Adaptación Fisiológica , Medios de Cultivo , Farmacorresistencia Bacteriana , Peróxido de Hidrógeno/análisis , Peróxido de Hidrógeno/metabolismo , Factores de Tiempo , Vibrio cholerae O1/crecimiento & desarrollo , Vibrio cholerae O1/metabolismoRESUMEN
The dynamics of the transition of V. cholerae into the uncultivable state in distilled, river and tap water, containing organic and inorganic components added, was studied. As additives, potassium nitrate, potassium phosphate, magnesium sulfate, ammonium chloride, lysine, alpha-ketoglutarate, succinic acid, catalase were used. The study of the influence of biotic factors on transition into the uncultivable state was carried out in the presence of one-celled green algae Scenedesmus quadricauda or infusoria Paramecium caudatum. The linear dependence of speed of transition into the uncultivable form on the concentration of cells was noted. The composition of the microcosmic medium was also found to have some influence on the speed of transition into the uncultivable form and on the reversibility of this process. The presence of organic substances, such as peptone solution or destroyed cells of phyto- and zooplankton, in the microcosmic medium prolonged the time of transition into the uncultivable form and produced a positive effect on the capacity of the population to reversion. In respect of live biotic components, no such dependence was found. Inorganic additives prolonged the time of transition into the uncultivable state, but did not promote reversion.
Asunto(s)
Vibrio cholerae/fisiología , Adaptación Fisiológica , Medios de Cultivo , Compuestos Inorgánicos , Compuestos Orgánicos/metabolismo , Paramecium caudatum/metabolismo , Peptonas/metabolismo , Scenedesmus/metabolismo , Factores de Tiempo , Vibrio cholerae/crecimiento & desarrollo , AguaRESUMEN
Testing the supernatants of ctx(+) strains of V. cholerae eltor and V. cholerae O139 on cell subcultures confirmed the possibility of the synthesis of hemolysin by V. cholerae under the condition of growing them in tripton medium lacking FeCl3. At the same time ctx(+) strains of V. cholerae of both serogroups retained, simultaneously with hemolysin production, their capacity for the synthesis of cholera toxin.
Asunto(s)
Proteínas Hemolisinas/biosíntesis , Vibrio cholerae O139/metabolismo , Animales , Proteínas Bacterianas , Línea Celular , Toxina del Cólera/biosíntesis , Medios de Cultivo , Medios de Cultivo Condicionados/metabolismo , Medios de Cultivo Condicionados/farmacología , Exotoxinas/biosíntesis , Compuestos Férricos , Humanos , Vibrio cholerae O139/crecimiento & desarrolloRESUMEN
A total of 20 ctx- and 16 ctx+ V. cholerae eltor strains, 20 ctx- and 22 ctx+ V. cholerae O139 strains were under study. Hemolytic activity was tested in modified Greig test with sheep, guinea pig and rabbit red blood cells. The comparative study of the hemolytic properties of V. cholerae O1 and O139 under different conditions of cultivation demonstrated their capacity of lysing sheep red blood cells (SRBC) irrespective of the presence of toxigenic properties. A wider spectrum of lytic activity of ctx- strains in Greig test with respect to red blood cells of different animals and the capacity of lysing SRBC, most resistant to the action of toxin, may be due to a considerably greater content of Hly+ clones in their population.
Asunto(s)
Toxina del Cólera , Hemólisis , Vibrio cholerae/inmunología , Animales , Proteínas Bacterianas , Cobayas , Proteínas Hemolisinas , Conejos , Ovinos , Especificidad de la Especie , Vibrio cholerae/patogenicidadRESUMEN
The hemolytic activity of ctx- and ctx+ V. cholerae, serogroups eltor and O39, in a medium free of FeCl3 was studied. During the cultivation in this medium, the strains of both V. cholerae serogroups proved to be capable of lysing sheep red blood cells in the Graig test, irrespective of the presence of ctx genes. The cultivation of V. cholerae ctx+ strains of both serogroups under such conditions facilitated the production of hemolysin with the same spectrum of lytic activity as hemolysin produced by ctx- strains.
Asunto(s)
Proteínas Hemolisinas/metabolismo , Vibrio cholerae/metabolismo , Animales , Animales Lactantes , Línea Celular , Cloruros , Toxina del Cólera/genética , Medios de Cultivo , Eritrocitos , Compuestos Férricos , Proteínas Hemolisinas/biosíntesis , Hemólisis , Humanos , Conejos , Ovinos , Vibrio cholerae/genética , Vibrio cholerae/patogenicidadRESUMEN
Experimental data on the comparative study of the invasive properties of vct+ Hly- and vct- Hly+ V. cholerae of serogroups 01 and 0139 are presented. Both vct- Hly+ and vct+ Hly- V. cholerae of serogroups 01 and 0139 have been shown to be capable of dissemination into internal organs. No differences in the dissemination of V. cholerae of different serogroups in both immunologically immature and mature experimental animals have been detected.
Asunto(s)
Cólera/microbiología , Vibrio cholerae , Animales , Animales Lactantes , Proteínas Bacterianas/genética , Cólera/inmunología , Modelos Animales de Enfermedad , Cobayas , Proteínas Hemolisinas/genética , Conejos , Receptores de Superficie Celular/deficiencia , Receptores de Superficie Celular/genética , Vibrio cholerae/genética , Vibrio cholerae/aislamiento & purificaciónRESUMEN
Experimental data on the comparative evaluation of the hemolytic activity of ctx+ Hly- and ctx- Hly+ V. cholerae, serogroups O1 and O139, in the process of their cultivation in different nutrient media are presented. The capacity of ctx+ V. cholerae of both serogroups cultivated under the conditions of iron deficiency, for the production of hemolysin capable of lyzing sheep red blood cells was shown. Hemolysin produced by ctx- strains of V. cholerae was synthesized under any conditions. The study of hemolysin preparations obtained from ctx- and ctx+ strains of V. cholerae, serogroups O1 and O139, revealed that they were biologically and immunologically similar.
Asunto(s)
Toxina del Cólera/metabolismo , Proteínas Hemolisinas/metabolismo , Vibrio cholerae/metabolismo , Animales , Animales Lactantes , Toxina del Cólera/inmunología , Cricetinae , Medios de Cultivo , Eritrocitos/inmunología , Proteínas Hemolisinas/inmunología , Humanos , Hierro , Conejos , Ovinos , Vibrio cholerae/patogenicidadRESUMEN
The influence of the preparation of hemo-cytolysin, obtained from V. cholerae non O1 strain P-11702 and inducing lysis of both red blood cells and V. cholerae cultures using a panel of indicator cultures for the detection of vibriocins, was studied. The set of indicator cultures contained 2 Shigella flexneri strains, 1 S. dysenteriae strain, 3 S. sonnei strains, 3 Escherichia coli strains and 2 V. cholerae strains, one of them being atypical. Hemo-cytolysin exhibited lytic activity with respect to S. dysenteriae, S. sonnei strains and 1 V. cholerae strain. i.e. to 4 out of 11 indicator strains. V. cholerae atypical strain proved to be resistant to the preparation in contrast to 33 V. cholerae typical strains, studied previously.
Asunto(s)
Bacteriólisis/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Proteínas Hemolisinas/farmacología , Shigella/efectos de los fármacos , Vibrio cholerae/efectos de los fármacos , Proteínas Bacterianas , Bacteriocinas/análisis , Escherichia coli/crecimiento & desarrollo , Shigella/crecimiento & desarrollo , Especificidad de la Especie , Vibrio cholerae/crecimiento & desarrolloRESUMEN
The influence of the preparation of hemocytolysin, obtained from V. cholerae non O1 (strain P-11702), on the growth of V. cholerae cells was studied. The study revealed that hemocytolysin is capable of inducing partial or complete bacterial lysis on the place of its application, depending on the protein load of the substance and the inoculation dose of microbes. Two electrophoretic fractions with molecular weights of 14 and 14.5 kD induced the cytolysis of sheep, rabbit, guinea-pig red blood cells and showed a bactericidal effect. The different sensitivity of Vct+ and Vct- strains of V. cholerae to different doses of hemocytolysin was studied.
Asunto(s)
Bacteriólisis/efectos de los fármacos , Proteínas Hemolisinas/farmacología , Vibrio cholerae/crecimiento & desarrollo , Proteínas Bacterianas , Vibrio cholerae/efectos de los fármacosRESUMEN
It was shown that plasmapheresis included in the complex program of intensive therapy of patients with acute pyo-septic lesions of the lungs and pleura allowed to effectively arrest manifestations of endotoxicosis and to improve results of treatment of this category of patients. Extracorporeal hemoxygenation of small volumes added to plasmapheresis at the expense of inclusion of dialyzer DIP-02-02 in the extracorporeal contour considerably reinforced the detoxicating effect of plasmapheresis at the expense of nonspecific biotransformation of toxins.
Asunto(s)
Oxigenación por Membrana Extracorpórea , Enfermedades Pulmonares/terapia , Plasmaféresis , Enfermedades Pleurales/terapia , Equilibrio Ácido-Base , Enfermedad Aguda , Lesión Renal Aguda/sangre , Lesión Renal Aguda/etiología , Lesión Renal Aguda/terapia , Análisis de los Gases de la Sangre , Terapia Combinada , Oxigenación por Membrana Extracorpórea/instrumentación , Oxigenación por Membrana Extracorpórea/métodos , Humanos , Fallo Hepático Agudo/sangre , Fallo Hepático Agudo/etiología , Fallo Hepático Agudo/terapia , Enfermedades Pulmonares/sangre , Enfermedades Pulmonares/complicaciones , Plasmaféresis/instrumentación , Plasmaféresis/métodos , Enfermedades Pleurales/sangre , Enfermedades Pleurales/complicaciones , Cuidados Preoperatorios/instrumentación , Cuidados Preoperatorios/métodos , Inducción de RemisiónRESUMEN
The authors have shown hyporegeneratory character of anemia developing after operations on the lungs followed by massive blood loss which was adequately compensated by conserved homologous blood. The transfusion of the autologous bone marrow prepared before operation results in a pronounced stimulation of production of erythrocytes, contributing to the quickest recovery of the circulating erythron.