Influence of Cultural Conditions on Cytotoxin Production by Salmonella enteritidis

The ability of Salmonella enteritidis strain 11013 to produce cytotoxic activity against Vero cells was determined under different cultural conditions. The toxin, which was not neutralizable with antiserum to Shiga toxin or Escherichia coli verotoxin-1 or verotoxin-2, was principally cell-associated and was produced primarily during the early stationary phase of growth. Trypticase soy broth was the best of three media evaluated for toxin production. Bacteria produced toxin in the range of pH 4.5 to 8.0 and at 12 to 42°C, with the optimal pH and temperature for toxin production at pH 7.0 and 37°C, respectively. Release of cellular cytotoxin into growth media was induced by growing salmonellae at extremes of pH (4.5 or 8.0) or at high incubation temperature (42°C). The Vero cell CD₅₀ of S. enteritidis lysates of cells grown under optimal conditions was a titer of 150 ± 50 per mg of lysate protein. Although the significance of ingesting preformed Salmonella cytotoxin in human disease is unknown, it can be implied from these results that toxin would not be produced in foods held refrigerated at ≤7°C or acidified at ≤pH 4.0.