This study aimed to determine the persistence of Salmonella Javiana, Listeria monocytogenes, and Listeria innocua in hydroponic nutrient solution (NS) at 15, 25, 30, and 37°C over a 21-day period to mimic time from seedling to mature lettuce. Bacteria were inoculated in modified Hoagland's NS at 106 CFU/ml and maintained at 15, 25, 30, and 37°C. Samples were collected at various time points and quantified. Data were analyzed using a mixed effect model to compare mean log CFU/ml obtained from each sampling point for all three bacteria at four different temperatures. Least mean squares were calculated to compare mean log CFU/ml. Tukey-Kramer honest significant difference test was used to compare mean values. At all temperatures, S. Javiana persisted in NS throughout the 21-day study period, compared to L. innocua and L. monocytogenes where persistence was limited to d 5 to d 14 and d 1 to d 14, respectively. Similarly, decimal reduction values (D-values) of S. Javiana indicated longer persistence in nutrient solution than L. innocua and L. monocytogenes at most temperatures. For instance, at 15°C and 25°C D-values for S. Javiana were estimated at 82 and 26 d, respectively, compared to D-values of 3.6 and ~3 d for L. monocytogenes. Data indicate that temperature has minimal effect on S. Javiana and thus may pose a greater risk during hydroponic production of leafy greens due to longer survival in NS when compared to Listeria spp. This study furthers the understanding of potential food safety risks associated with hydroponic systems and may aid in developing management strategies to reduce foodborne outbreaks, fresh produce recalls, and economic losses.