Biophysiological and factorial analyses in the treatment of rural domestic wastewater using multi-soil-layering systems.

Multi-soil-layering (MSL) system was developed as an attractive alternative to traditional land-based treatment techniques. Within MSL system, the environmental cleanup capability of soil is maximized, while the soil microbial communities may also change during operation. This study aimed to reveal the nature of biophysiological changes in MSL systems during operation. The species diversity in soil mixture blocks was analyzed using Illumina HiSeq sequencing of the 16S rRNA gene. The interactive effects of operating factors on species richness, community diversity and bacteria abundance correlated with COD, N and P removal were revealed through factorial analysis. The results indicated the main factors, aeration, bottom submersion and microbial amendment, had different significant effects on microbial responses. The surface area and porosity of zeolites in permeable layers decreased due to the absorption of extracellular polymeric substances. The findings were applied for the design and building of a full-size MSL system in field and satisfied removal efficiency was achieved. The results of this study can help better understand the mechanisms of pollutant reduction within MSL systems from microbial insights. It will have important implications for developing appropriate strategies for operating MSL systems with high efficiency and less risks.