This research comparatively investigates the biotoxicity of landfill leachate effluent from acclimatized and non-acclimatized sludge two-stage activated sludge (AS) systems. Both AS systems were operated with two leachate influent concentrations: moderate (condition 1) and elevated (condition 2). The biotoxicity of AS effluent of variable concentrations (10, 20, and 30 % (v/v)) was assessed by the mortality rates of common carp (Cyprinus carpio) and glutathione-S-transferase (GST) enzyme activity. The treatment efficiency of the acclimatized sludge AS system for organic and inorganic compounds and nutrients (BOD, COD, TKN, NH₄⁺, PO₄^3-) were 75 – 96% under condition 1 and 79 – 93% under condition 2. The non-acclimatized sludge AS system achieved the treatment efficiency of 70 – 91% under condition 1 and 66 – 90% under condition 2. The acclimatized sludge AS system also achieved higher biodegradation of trace organic compounds, especially under condition 1. The effluent from acclimatized sludge AS system was less toxic to the common carp, as evidenced by lower mortality rates and higher GST activity. The findings revealed that the acclimatized sludge two-stage AS system could be deployed to effectively treat landfill leachate with moderate concentrations of compounds and trace organic contaminants. The acclimatized sludge AS is an efficient wastewater treatment solution for developing countries with limited technological and financial resources.