Inhibition of Salmonella enterica by plant-associated pseudomonads in vitro and on sprouting alfalfa seed.

Foodborne illness due to the consumption of contaminated raw or lightly cooked sprouts is a continuing food safety concern. In this study, we tested several plant-associated pseudomonads for their ability to inhibit the growth of Salmonella enterica both in vitro and in situ. An agar spot bioassay method was used with three different media. Only Pseudomonas fluorescens 2-79 produced clear zones of inhibition when tested against five serovars of S. enterica, and activity was dependent on media type and serovar. The antibiosis by derivative strains of P. fluorescens 2-79 defective in the production of phenazine-1-carboxylic acid and fluorescent siderophore was not reduced, indicating that these known antimicrobial metabolites were not responsible for the inhibition observed in our studies. However, mutants defective in the regulatory gene gacS (global antibiotic and cyanide control) were severely reduced in inhibitory activity. In tryptic soy broth, the control cultures of a cocktail of S. enterica strains reached approximately 10 log CFU/ml by 24 h but, when coinoculated with P. fluorescens 2-79, reached only approximately 5 log CFU/ml. The addition of P. fluorescens 2-79 to the seed soak water prior to the germination of alfalfa seed previously inoculated with a cocktail of S. enterica strains led to an average reduction of 5 log CFU/g at 6 days of sprouting without an adverse effect on sprout yield or appearance. Time course studies indicated that S. enterica outgrowth was controlled on days 1 through 6 of sprouting. Competitive exclusion as a potential food safety intervention for seed sprouts merits further study.