The Combination of Lactate and Diacetate Synergistically Reduces Cold Growth in Brain Heart Infusion Broth across Listeria monocytogenes Lineages

Combinations of organic acids are often used in ready-to-eat foods to control the growth of Listeria monocytogenes during refrigerated storage. The purpose of this study was to quantitatively assess synergy between two organic acid growth inhibitors under conditions similar to those present in cold-smoked salmon, and to assess the effect of evolutionary lineage on response to those growth inhibitors. Thirteen strains of L. monocytogenes, representing lineages I and II, were grown at 7°C in broth at pH 6.1 and 4.65% water-phase NaCl, which was supplemented with 2% potassium lactate, 0.14% sodium diacetate, or the combination of both at the same levels. Our data suggest that lineages adapt similarly to these inhibitors, as the only significant growth parameter difference between lineages was a minor effect (± 0.16 day, P = 0.0499) on lag phase (λ). For all strains, lactate significantly extended λ, from 2.6 ± 0.4 to 3.8 ± 0.5 days (P < 0.001), and lowered the maximum growth rate (μ_max) from 0.54 ± 0.06 to 0.49 ± 0.04 log(CFU/ml)/day (P < 0.001), compared with the control. Diacetate was ineffective alone, but in combination with lactate, synergistically increased λ to 6.6 ± 1.6 days (P < 0.001) and decreased μ_max to 0.34 ± 0.05 log(CFU/ ml)/day (P < 0.001). Monte Carlo simulations provided further evidence for synergy between diacetate and lactate by predicting signficantly slower growth to nominal endpoints for the combination of inhibitors. This study shows potassium lactate and sodium diacetate have significant synergistic effects on both λ and μ_max of L. monocytogenes at refrigeration temperature in broth, and justifies combining these inhibitors, at effective levels, in food product formulations.