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Hydroponic cultivation of lettuce is an increasingly popular sustainable agricultural technique. However, <i>Escherichia coli</i>, a prevalent bacterium, poses significant concerns for the quality and safety of hydroponically grown lettuce. This study aimed to develop a growth model for <i>E. coli</i> and background microflora in hydroponically grown lettuce. The experiment involved inoculating hydroponically grown lettuce with <i>E. coli</i> and incubated at 4, 10, 15, 25, 30, 36 °C. Growth models for <i>E. coli</i> and background microflora were then developed using Origin 2022 (9.9) and IPMP 2013 software and validated at 5 °C and 20 °C by calculating root mean square errors (RMSEs). The result showed that <i>E. coli</i> was unable to grow at 4 °C and the SGompertz model was determined as the most appropriate primary model. From this primary model, the Ratkowsky square root model and polynomial model were derived as secondary models for <i>E. coli</i>-R168 and background microflora, respectively. These secondary models determined that the minimum temperature (T_min) required for the growth of <i>E. coli</i> and background microflora in hydroponically grown lettuce was 6.1 °C and 8.7 °C, respectively. Moreover, the RMSE values ranged from 0.11 to 0.24 CFU/g, indicating that the models and their associated kinetic parameters accurately represented the proliferation of <i>E. coli</i> and background microflora in hydroponically grown lettuce.