Subject(s)
Transfer Factor/analysis , Amino Acids/isolation & purification , Chromatography, Gel , Chromatography, Thin Layer , Guanosine/isolation & purification , Humans , Hypoxanthines/isolation & purification , Peptides/isolation & purification , Uracil/isolation & purification , Uridine/isolation & purificationABSTRACT
Immunological methods have been used to visualize peroxidase and dehydrogenase activities of substrains of BCG mycobacteria. Immunodiffusion (ID) and immunoelectrophoresis (IE) in agar gel and two-dimensional immunoelectrophoresis (2D-IE) in agarose gel were employed. In all these tests part of the ID, IE, and 2D-IE preparations were stained with Amido Black 10B (AB 10B) to visualize the total number of precipitin lines and part were stained at the same time with histochemical methods for vizualization of peroxidase and dehydrogenase activities.
Subject(s)
BCG Vaccine , Immunologic Techniques , Mycobacterium bovis/enzymology , Immunodiffusion , Immunoelectrophoresis , Immunoelectrophoresis, Two-Dimensional , Methods , Oxidoreductases/metabolism , Peroxidases/metabolismABSTRACT
The usefulness of the method of two-dimensional immunoelectrophoresis (2D-IEP) in comparative studies and characterization of the antigenic spectra of various BCG substrains have been estimated. The BCG substrains used were: substrain BCG-Rio de Janeiro, substrain BCG-Poland, form-rough (R) and substrain BCG-Poland, form-smooth (S). The 2D-IEP technique was found to give characteristic immunoelectrophoretic patterns of antigenic composition of the different BCG substrains. The number of precipitin peaks was usually over 20, that is more than detected by conventional immunoprecipitation techniques such as immunodiffusion and immunoelectrophoresis in agar gel (ID, IE).
Subject(s)
Antigens, Bacterial/analysis , BCG Vaccine , Mycobacterium bovis/immunology , Immunoelectrophoresis, Two-DimensionalABSTRACT
The influence of iron concentration in Sauton's medium solidified with agar on the type of colonies of BCG-Poland substrains, BCG-Rio de Janeiro, BCG-France, BCG-Denmark and BCG-Japan substrains has been examined. Of all the studied BCG substrains only the BCG-Poland substrain formed rough (R) and smooth (S) colonies. In the investigated substrains rough colonies became smaller with the decrease of iron concentration but they retained their characteristic surface roughness. The smooth colonies which in the same conditions appeared only in BCG-Poland substrain did not display such dependency on iron concentration. When the incubation period was prolonged secondary rough colonies appeared among the smooth ones, regardless of the iron concentration in the medium.
Subject(s)
BCG Vaccine , Iron/pharmacology , Mycobacterium bovis/drug effects , Culture Media , Mycobacterium bovis/cytology , Mycobacterium bovis/growth & developmentABSTRACT
The dissociation of colonies of the substrain BCG-Poland on Löwenstein-Jensen medium has been studied. It had been stated previously (Lipinska and Rzucidlo, 1974) that BCG-Poland substrain undergoes a process of dissociation which manifests itself in the appearance of smooth colonies alongside typical rough colonies. Attempts have been made to obtain pure culture of both colony types. Pure cultures composed of smooth or rough colonies respectively were obtained. The morphology and diameter of the smooth colonies depended on the size of the seed inoculum spread on the medium.
