RESUMO
A thermolabile extracellular entero-cytolysin (EC) from Vibrio cholerae non-O1 was purified by ammonium sulphate fractionation, DE-52 cellulose ion exchange chromatography, gel-filtration on Ultrogel AcA-44 and high performance liquid chromatography on a Mono Q. The purified EC had a molecular weight of 63 kD and an isoelectric point of 6.2. It was not inactivated by cholesterol or 5,5'-dithio-bis(2-nitrobenzoic acid), nor activated by dithiothreitol. EC had no immunological cross-reactivity with cholera toxin. The EC caused fluid accumulation in the intestines of infant rabbits, death of mice by intravenous injection, and increased vascular permeability in the paw oedema test in mice. V. cholerae non-O1 EC lysed erythrocytes from various species and cultured cells (CHO, L-929, L-41, HEp-2, Vero, MDCK and BHK-21). In contrast to cholera toxin, EC caused crude destruction of target cells. The EC caused hemolysis by a colloid-osmotic mechanism due to the formation of hydrophilic pores of 1.8-2.0 nm diameter in the cell membrane. This EC also was able to open pores in lipid membranes. The induced channels were anion-selective and had a diameter of 1.8-2.0 nm.
Assuntos
Citotoxinas/toxicidade , Enterotoxinas/toxicidade , Vibrio cholerae/química , Animais , Permeabilidade Capilar/efeitos dos fármacos , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Cromatografia Líquida , Citotoxinas/química , Citotoxinas/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Enterotoxinas/química , Enterotoxinas/isolamento & purificação , Membrana Eritrocítica/efeitos dos fármacos , Hemólise , Canais Iônicos/efeitos dos fármacos , Ponto Isoelétrico , Dose Letal Mediana , Bicamadas Lipídicas , Camundongos , Peso Molecular , Coelhos , Vibrio cholerae/patogenicidadeRESUMO
Enzyme immunoassay was used in detection of V. cholerae cytolysin. Conjugate of immunoglobulins to purified cytolysin with horseradish peroxidase was used, obtained by the periodate technique. The method sensitivity is 2 ng/ml of purified cytolysin. The results of hemolytic activity measurements in supernatants of 40 V. cholerae strains and enzyme immunoassay findings were in high correlation.
Assuntos
Citotoxinas/análise , Vibrio cholerae/análise , Técnicas ImunoenzimáticasRESUMO
A new method for isolation and purification of Vibrio cholerae non-01 cytolysin has been elaborated. It includes the steps of concentration by ammonium sulphate, ion exchange chromatography on DE-52-cellulose, gel filtration via ultragel AsA-44 and chromatography on Mono Q. Cytolysin is shown to be a 60 kD and pI 6.2 protein. It is not inactivated by thiol-restoring agents and induces the production of precipitating antibodies. In contrast to choleragen the protein cytolytically affects erythrocytes and cells of the line L-929. It also possesses the lethal and oedematous effect and demonstrates the enteropathogenic effect on nursing rabbits.
Assuntos
Citotoxinas/isolamento & purificação , Vibrio cholerae/metabolismo , Animais , Cromatografia por Troca Iônica , Citotoxinas/química , Citotoxinas/toxicidade , Eletroforese em Gel de Poliacrilamida , Imunodifusão , Dose Letal Mediana , Camundongos , CoelhosRESUMO
Subdivision of V. cholerae 01 into toxins on the basis of the whole complex of signs characteristic of this species does not make it possible to judge on their epidemic importance and to use the data on identification of V. cholerae for solving practical problems. Classification of V. cholerae by their capacity for producing toxin (choleragen and hemolysin of type 1, subtype beta) removes these difficulties.
Assuntos
Cólera/microbiologia , Surtos de Doenças , Vibrio cholerae/patogenicidade , Animais , Animais Lactentes , Toxina da Cólera/análise , Toxina da Cólera/biossíntese , Toxina da Cólera/classificação , Toxina da Cólera/toxicidade , Cobaias , Hemólise/efeitos dos fármacos , Fenótipo , Coelhos , Ovinos , Vibrio cholerae/classificaçãoAssuntos
Febre/microbiologia , Infecções por Pasteurella/microbiologia , Pasteurella/isolamento & purificação , Sepse/microbiologia , Adolescente , Testes de Aglutinação/métodos , Feminino , Febre/sangue , Febre/diagnóstico , Humanos , Infecções por Pasteurella/sangue , Infecções por Pasteurella/diagnóstico , Sepse/sangue , Sepse/diagnósticoAssuntos
Pasteurella/classificação , Animais , Cobaias , Humanos , Métodos , Camundongos , Pasteurella/patogenicidade , CoelhosRESUMO
Variation diagrams were proposed to be used for the analysis of the published data on the study of Vibrio cholerae sensitivity to tetracycline. The diagrams provided systematization of dissimilar data of the experimental studies and a unique system of estimation was created. The analysis of the systematized data provided dividing of the Vibrio cholerae strains tested into 4 groups differing in the levels of their sensitivity to tetracycline. The groups were independent of the Vibrio cholerae biotypes and the place and period of the strain isolation.
Assuntos
Tetraciclina/farmacologia , Vibrio cholerae/efeitos dos fármacos , Relação Dose-Resposta a Droga , Resistência Microbiana a Medicamentos , Concentração de Íons de Hidrogênio , Índia , Testes de Sensibilidade Microbiana , Estatística como AssuntoRESUMO
Prolonged (for a period of one year) isolation of cholera vibrios from the sewage of a bath-house was observed. Sewage proved to serve as a favourable medium for preservation and reproduction of cholera vibrios. The data obtained pointed to the capacity of El Tor cholera vibrio to exist outside the human organism for a long time.
