Evidence for phospholipase B activity in Fusobacterium necrophorum cultures and its association with hemolysin/leucocidin activities

Phospholipase B (PLB) activity was present in Fusobacterium necrophorum cultures and it correlated closely with virulence and co-purified with the hemolysin/leucocidin activities. All three activities were associated with a large molecule or molecular complex (6 X 10(2)-2 X 10(3) kDa) exhibiting varying degrees of aggregation. These were present mainly in the culture medium and to a lesser extent in cell sonicates. The PLB and toxin activities were sensitive to heat, dissociating agents, proteolyric enzymes, prolonged purification regimes, freeze-drying and repeated freeze-thawing. The toxin(s) were stable over a broad range of pH, did not require divalent ions or reducing agents and could be kept for several months as an ammonium sulfate precipitate at 4 degrees C, or stored as a concentrated liquid in the presence of proteolytic inhibitors at -20 degrees C.