Characterization of the hemolysin produced by haemophilus pleuropneumoniae

Production and characterization of Haemophilus pleuropneumoniae hemolysin were investigated by using 5 serotype 2 strains. The hemolysin was produced in chicken meat-infusion broth medium both in stationary and in shaking cultures. In stationary culture, hemolytic activity against horse RBC reached maximum at postincubation day 5 (at the late stage of stationary phase), and the activity was maintained at the same level for 2 days thereafter. The hemolytic activity of shaking culture reached a maximum at postincubation hour 9 (at the early stage of logarithmic-growth phase), gradually decreased, and disappeared at postincubation day 2. The hemolysin was shown to be an extracellular product of the bacterial cells. The RBC of horses, rabbits, and sheep were highly susceptible to the hemolysin, and those of pigs and guinea pigs were less susceptible, whereas RBC of 60-day-old chicks were not susceptible. The hemolysin was not inactivated by autoclaving at 121 C for 2 hours, and by treatments with formalin, trypsin, or pronase. The presence of calcium or magnesium ions did not change the activity, whereas iodoacetic acid significantly reduced the activity.