SALMONELLA ENTERICA STRATEGIES FOR PERSISTENCE ON TOMATO (SOLANUM LYCOPERSICUM) AND SEROVAR DYNAMICS IN SURFACE AND RECLAIMED WATER

Abstract

While select aspects of Salmonella enterica subspecies enterica persistence in agricultural matrices have been illustrated, serovar specific survival strategies in surface water, transmission, and persistence on plants are multifaceted and remain only partially examined. In the present work, we utilized an interdisciplinary approach to illustrate novel mechanisms by which S. enterica may adapt to plants as an alternative host. Furthermore, we leveraged the wealth of diversity in S. enterica serovars to investigate specific dynamics and drivers of persistence in water and transfer onto produce crops. Through biochemical, gene expression, and plant challenge assays of both tomato (Solanum lycopersicum) vegetative and fruit organs, we found that plant-derived NO was generated in response to S. Newport recognition. Furthermore, bacterial gene expression on both leaves and fruit was indicative of adaptation to a novel environment including upregulation in NO detoxification machinery, indicating plant-derived NO as a novel bacterial stress. NO tolerance of various S. enterica was then evaluated to investigate drivers of “produce associated’ S. enterica adaptation to the plant niche. We identified that plant derived NO can negatively affect titers of all S. enterica serovars tested and that serovar specific tolerance to NO in vitro was apparent in a concentration and exposure time dependent manner. Finally, the survival of various S. enterica in surface and reclaimed water was investigated while evaluating the potential for transition to viable but non-culturable (VBNC) organisms. Furthermore, surface water used for irrigation, a common water environment for S. enterica, was investigated as a priming reservoir for various S. enterica serovars for enhanced transmission onto tomato crops. Persistence in water included VBNC subpopulations and was driven by water type. Transfer success onto tomato was driven by serovar, and prolonged incubation in water increased the transfer ability of serovars that initially transferred poorly onto tomato. Finally, attachment to polystyrene and water oxidation-reduction potential were identified as possible indicators of foodborne pathogen transfer success onto tomato. Moving forward, a greater understanding of the environmental queues used by S. enterica subspecies enterica responding to the agricultural environment will aid researchers in developing S. enterica targeted on-farm management techniques to ensure safe yet sustainable fresh produce cultivation practices.