Effects of henna plant biomass (stem) packed in an up-flow anaerobic bio-filter (UAF) on an azo dye (AO7) removal were investigated. AO7 removal, sulfanilic acid (SA) formation, and pseudo first-order kinetic constants for these reactions (k(AO7) and k(SA)) were higher in the henna-added UAF (R2) than in the control UAF without henna (R1). The maximum k(AO7) in R1 and R2 were 0.0345 and 0.2024 cm(-1), respectively, on day 18; the corresponding molar ratios of SA formation to AO7 removal were 0.582 and 0.990. Adsorption and endogenous bio-reduction were the main AO7 removal pathways in R1, while in R2 bio-reduction was the dominant. Organics in henna could be released and fermented to volatile fatty acids, acting as effective electron donors for AO7 reduction, which was accelerated by soluble and/or fixed lawsone. Afterwards, the removal process weakened over time, indicating the demand of electron donation and lawsone-releasing during the long-term operation of UAF. (C) 2015 Elsevier B.V. All rights reserved.