Addressing the global challenge of uranium (U)-contaminated groundwater requires innovative bioremediation strategies. This study investigates Desulfovibrio desulfuricans, a neutrophilic and mesophilic sulfate-reducing bacteria (SRB) strain optimized for low-temperature (15 °C) and acidic (initial pH 4) conditions, to validate its bioaugmentation potential for uranium decontamination in groundwater. Our research aimed to assess its efficacy in treating U-contaminated groundwater and elucidate the optimal growth conditions for this strain in acidic and sulfate-enriched environments. We found that D. desulfuricans was phylogenetically distinct from the native microbial community in acidic U-contaminated groundwater, while it maintained appreciable activity in sulfate reduction under contaminated groundwater conditions after accumulation. Acid-tolerant D. desulfuricans removed 75.87 % of uranium and 30.64 % of sulfate from acidic U-contaminated groundwater (pH 4.0) at 15 °C within 14 days. Furthermore, we explored the optimal sulfate concentration for bacterial growth, which was found to be 2000 mg/L, and an elevated Fe2+ concentration from 100 to 1000 mg/L increasingly stimulated sulfate-reducing activity. These findings provide a novel insight into the application of neutrophilic and mesophilic SRB in bioremediation of acidic and low-temperature groundwater after accumulation and underscore the feasibility of bioremediation by using exogenously pure SRB.