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The threat posed by water pollutants to aquatic ecosystems and human health cannot be overlooked, and the assessment of the toxicity of these contaminants is paramount to understanding their risks and formulating effective control measures. Luminescent bacteria-based assays, as a vital tool in evaluating contaminant toxicity, encounter a challenge in ensuring accuracy due to the phenomenon of “Hormesis” exhibited by pollutants towards biological entities, which may skew toxicity assessments. This study elucidated the specific effects of pollutants on luminescent bacteria at different concentrations, used modeling to characterize the effects and predict their toxicity trends, and explored the applicable concentration ranges for different pollutants. Research revealed that six typical pollutants, namely PAHs, endocrine disruptors, antibiotics, pesticides, heavy metals, and phytosensory substances, could promote the luminescence intensity of luminescent bacteria at low concentrations, and the promotional effect increased and then decreased. However, when the concentration of the substances reached a certain threshold, the effect changed from promotional to inhibitory, and the rate of inhibition was directly proportional to the concentration. The EC50 values of six types of substances to luminescent bacteria is as follows: endocrine disruptors > pesticides > antibiotics > heavy metals > polycyclic aromatic hydrocarbons > chemosensory agents. The effect curves were further fitted using the model to analyze the maximum point of the promotion of luminescence intensity by different substances, the threshold concentration, and the tolerance of luminescent bacteria to different substances. The maximum promotion of bacterial luminescence intensity was 29% for Bisphenol A at 0.005 mg/L and the minimum threshold concentration of chromium was 0.004 mg/L, and the maximum bacterial tolerance to erythromycin is 6.74. In addition, most of the current environmental concentrations had a positive effect on luminescent bacteria and may still be in the range of concentrations that promote luminescence as the substances continue to accumulate. These findings will enhance the accuracy and comprehensiveness of toxicity assessments, thereby facilitating more informed and effective decision-making in the realms of environmental protection and pollution management.