Unraveling the capability of graphene nanosheets and γ-Fe2O3 nanoparticles to stimulate anammox granular sludge.

In this study, we investigated the potentials of nanomaterials to enhance anaerobic ammonium oxidation (anammox) process, in terms of nitrogen removal, microbial enrichment, and activity of key enzymes. Graphene nanosheets (GNs) and γ-Fe2O3 nanoparticles (NPs) were selected due to their catalytic functions as conductive material and electron shuttles, respectively. The obtained results revealed that the optimum dosage of GNs (10 mg/L) boosted the nitrogen removal rate (NRR) by 46 ± 3.1% compared to the control, with maximum NH4+-N and NO2--N removal of 86.5 ± 2.7% and 97.1 ± 0.5%, respectively. Moreover, hydrazine dehydrogenase (HDH) enzyme activity was augmented by 1.1-fold when using 10 mg/L GNs. The presence of GNs promoted the anammox granulation via enhancement of hydrophobic interaction of extracellular polymeric substances (EPS). Regarding the use of γ-Fe2O3 NPs, 100 mg/L dose increased NRR by 55 ± 3.8%; however, no contribution to HDH enzyme activity and a decrease in EPS compositions were observed. Given that the abiotic use of γ-Fe2O3 NPs further resulted in high adsorption efficiency (~92%), we conclude that the observed promotion due to γ-Fe2O3 NPs was mainly abiotic. Moreover, the 16S rRNA analysis revealed that the relative abundance of genus C. Jettenia (anammox related bacteria) increased from 11.9% to 12.3% when using 10 mg/L GNs, while declined to 8.3% at 100 mg/L γ-Fe2O3 NPs. Eventually, nanomaterials could stimulate the efficiency of anammox process, and this promotion and associated mechanism depend on their dose and composition.