Impact of benzalkonium chloride on anaerobic granules and its long-term effects on reactor performance

This study assessed the inhibitory and performance-degrading effects induced by the cationic surfactant benzalkonium chloride (BAC) on anaerobic granules during the long-term operation of a laboratory-scale expanded granular sludge bed (EGSB) reactor. To address the critical scientific problem of how BAC affects the efficiency of EGSB reactors, this research uniquely evaluated the long-term stress response to BAC by systematically comparing continuous and discontinuous inhibitor exposure scenarios. The novel comparison demonstrated that inhibitor concentration is of minor relevance compared to the biomass-specific cumulative inhibitor load in the reactor. After exceeding a critical biomass-specific cumulative inhibitor load of 6.1-6.5 mg BAC/g VS, continuous and discontinuous exposure to BAC caused comparable significant deterioration in reactor performance, including accumulation of volatile fatty acids (VFA), decreased removal efficiency, reduced methane production, as well as the wash-out, flotation, and disintegration of anaerobic granules. BAC exposures had a more detrimental effect on methanogenesis than on acidogenesis. Moreover, long-term stress by BAC led to an inhibition of protein production, resulting in a decreased protein-to-polysaccharide ratio of extracellular polymeric substances (EPS) that promoted destabilizing effects on the granules. Finally, hydrogenotrophic methanogenesis was triggered. Reactor performance could not be restored due to the severe loss of granular sludge.