Performance of loach to reduce clogging matter in vertical-flow constructed wetlands: Hydraulic evaluation and metabolic processes

Extracellular polymeric substances (EPS) secreted by microorganisms cause constructed wetlands (CWs) clogging, which is a major challenge for the long-term stable operation of CWs. This study provided a comprehensive and in-depth study of the hydraulic evaluation and metabolic processes of loach (Misgurnus anguillicaudatus) in mitigating clogging. The results showed that the loach reduced the clogging content in the substrate layer and improved the hydraulic performance of the system. The loach system increased the initial porosity in the sand layer by 11.52 %, 25.11 %, and 28.15 %, and the initial porosity in the gravel layer by 15.49 %, 15.85 %, and 13.83 %, respectively. Loach also increased the actual and theoretical hydraulic retention time (HRT), improved the connectivity between different layers, and reduced the "dead zone" area of the system. The analysis of EPS components revealed that the daily EPS removal amount of each loach was 36.64 mg L-1, in which S-EPS, LB-EPS, and TB-EPS were 21.80, 8.02, and 6.82 mg L-1d-1, respectively. Direct feeding and the indirect coordination with microorganisms effectively reduced the contents of polysaccharides and proteins in EPS, and increased the proportion of humic acid and nucleic acid. The clogging characteristics analysis also confirmed that loach decreased tyrosine and tryptophan fluorophores groups and reduced the proportion of protein or peptide substances in the macromolecules in EPS. In addition, loach increased the activities of dehydrogenase, fluorescein diacetate, and the relative abundance of Chlorobiaceae in the sand layer, which was conducive to the hydrolysis of clogging matters with the metabolic activity of microorganisms. Loach increased the relative abundance of denitrifying microorganisms such as Aeromonadaceae, Rhodocyclaceae, and Hydrogenophilaceae in the gravel layer, which enhanced the denitrification process of the system and was conducive to pollutant transformation and clogging matter degradation. In conclusion, this study proposes a novel, eco-friendly strategy for preventing CW clogging in wetland animals.