Formation and Microbial Composition of Biofilms in Drip Irrigation System under Three Reclaimed Water Conditions

As the second source of water for cities, reclaimed water (RW) has become an effective solution to the problem of water scarcity in modern agriculture. However, the formation of biofilm in an RW distribution system seriously affects the performance of the system and has become a technical challenge in RW utilization. In this study, we first showed that several water quality parameters, including five-day biochemical oxygen demand (BOD₅), total bacteria count (TB), total nitrogen (TN), and Cl⁻ were the main factors affecting biofilm accumulation in the drip irrigation system (DIS), with the correlation coefficient averaging above 0.85. Second, after 392 to 490 h of system operation, the total biomass and extracellular polymer (EPS) accumulation rate of biofilms increased to a maximum of 0.72 g/m²·h and 0.027g/m²·h, respectively, making this time point a critical point for controlling biofilm accumulation and clogging of the system. Third, we examined changes in biofilm microbial composition over time on Illumina’s MiSeq platform. High throughput sequencing data showed that bacterial community structure and microbial network interaction and modularity changed significantly between 392 and 490 h, resulting in maximum microbial diversity and community richness at 490 h. Spearman correlation analyses between genera revealed that Sphingomonas and Rhodococcus promote biofilm formation due to their hydrophobicity, while Bacillus, Mariniradius, and Arthronema may inhibit biofilm formation due to their antagonistic effects on other genera. In conclusion, this work has clarified the accumulation process and compositional changes of biofilms in agriculture DIS under different RW conditions, which provides a basis for improving RW utilization efficiency and reducing system maintenance costs.