a b s t r a c tNatural estrogens (NEs) discharged from wastewater treatment plants (WWTPs) have drawn great attention because of their potential risks to aquatic ecosystems. However, neglect of the conjugated natural estrogens (C-NEs) has caused large discrepancies among different studies on the removal of NEs in WWTPs. The present work investigated the transformation and fate of three NEs and six corresponding C-NEs along wastewater treatment processes. The removal efficiencies of the target estrogens (i.e., NEs and C-NEs) and their correlations with the operational parameters were determined over a twelve-month monitoring period at a typical WWTP adopting a combined bio-treatment process (i.e., anaerobic/anoxic/oxic process followed by a moving-bed biofilm reactor). The concentration variations of the target estrogens along the treatment processes were examined to differentiate the transformation and fate of NEs and C-NEs. Moreover, lab-scale experiments were conducted to clarify the removal pathways of C-NEs in the bio-treatment process. Results indicate that both NEs and C-NEs could pass through the treatment processes, thus being frequently detected in the effluent and excess sludge. The aqueous removal efficiencies of NEs and C-NEs were significantly correlated with the sludge retention time and temperature, respectively. C-NEs were more persistent than NEs, so considerably high conjugated ratios (13.5e100.0%) were detected in the effluent. Sulfate conjugates presented a lower adsorption affinity to sludge and a slower hydrolysis rate than glucuronide conjugates, which makes the former more recalcitrant to biodegradation. This study highlights the challenge on the elimination of NEs, particularly their conjugates, by wastewater treatment processes.