The management of produced water (PW) generated during oil and gas operations requires effective treatment and comprehensive chemical and toxicological assessment to reduce the environmental risks associated with reuse or discharge. This study evaluated a treatment train that included a low-temperature thermal distillation pilot system followed by granular activated carbon (GAC) and zeolite post-treatment for processing hypersaline Permian Basin PW. Our study provides a unique and comprehensive assessment of the treatment efficiency considering a targeted chemical scheme together with whole effluent toxicity (WET) tests across four trophic levels regarding aquatic critical receptors of concern (ROC): Raphidocelis subcapitata, Vibrio fischeri, Ceriodaphnia dubia, and Danio rerio. The distillate from the thermal distillation process met various numeric discharge standards for salinity and major ions. However, it did not meet toxicity requirements established by the United States National Pollutant Discharge Elimination System program. Subsequent post-treatment using GAC and zeolite reduced the concentration of potential stressors, including volatile organics, NH3, Cd, Cr, Zn, and Mn in the final effluent to below detection limits. This resulted in a consistent toxicity reduction across all WET tests, with no observable adverse effects for R. subcapitata, C. dubia, and D. rerio (no observed effect concentration >100%), and V. fischeri effects reduced to 19%. This study realizes the feasibility of treating PW to non-toxic levels and meeting reuse and discharge requirements. It underscores the importance of implementing integrated treatment trains to remove the contaminants of concern and provides a systematic decision framework to predict and monitor environmental risks associated with PW reuse.