Testing multiple variables on the growth of a mixed inoculum of Salmonella strains using gradient plates.

Six Salmonella strains were grown on two-dimensional sodium chloride-pH and temperature-pH gradient plates. Using image analysis the results were expressed in the form of three-dimensional wire frame graphs. On the temperature-pH gradient plates the optimum growth range was 20-30 degrees C and the minimum pH for visible growth was ca. pH 4, except for strain S. typhimurium CRA63 which grew over a narrower temperature and pH range. On the NaCl-pH gradient plates (whose NaCl gradient began at 2.75% (w/v) NaCl) the maximum concentration of salt at which growth was visible varied from 3.9% to 6.0%, and the minimum pH at which growth was observed varied from pH 4.7 to 5.4 according to the strain used. The incorporation of 0.02% (w/v) sodium nitrite reduced the maximum salt concentration and increased the minimum pH at which the strains could grow. The strains were combined and used in a mixed inoculum on NaCl-pH gradient plates containing 6 different concentrations of NaNO2 incubated at 6 different temperatures. Comparison of the data from the mixed inoculum with data from individual strains showed that, apart from one case, the mixed inoculum represented the extremes of the growth domains of the individual strains. The effect of NaNO2 on the ability of the strains to grow at different pH, NaCl concentrations and temperatures, was more clearly shown by subtracting the data of plates containing NaNO2 from the data of plates without NaNO2.