Obtaining specific components for the manufacture of erythrocyte diagnosticum for clostridial infections in animals

The article presents the results of studies on obtaining specific components for staging the indirect hemagglutination assay in animals with anaerobic enterotoxemia. For the manufacture of erythrocyte diagnosticum, toxoids and soluble antigens intended for sensitization of formalized ram erythrocytes, as well as positive and negative blood serum of rabbits to antigens of bacteria Cl. perfringens serotypes A, B, C, D, E were obtained. Anatoxins were obtained by inoculation of industrial strains of Cl. perfringens N 28 (type A), LD-1 (type B), N 392 (type C), N 213 (type D), N 342 (type E) in liquid meat-liver-casein medium under vaseline oil and cultivating them for 10 hours at a temperature of 37-38 °C, followed by inactivation of the culture with formalin, purification and concentration of the toxoid by precipitation with ammonium sulfate. Somatic soluble antigen was obtained by destroying Cl. perfringens bacteria grown on beef-extract agar with ultrasound on a «Bandelin GM3100» disintegrator at a frequency of 20 kHz ± 500 Hz for 10 minutes at a temperature of 4 °C. After centrifugation of the suspension at 3000 rpm for 30 minutes, the supernatant was used as an antigen for sensitization of ram erythrocytes. Positive serum to the antigens of bacteria Cl. perfringens was obtained on rabbits with a live weight of 3-3.5 kg by three intravenous immunizations with a composite mixture of toxoids and antigens with an interval of 4 days. Control negative serum obtained from intact rabbits with a live weight of 3-3.5 kg.