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Long-distance water transfer projects are important for water allocation. To enhance our understanding of biofilm growth and changes in water quality during raw water transfer, raw water through a long-distance non-full culvert at flow rates of 1.4−2.0 m/s was studied. The results revealed that: (1) the biofilm total cell count (TCC) and heterotrophic plate count (HPC) were the highest at a flow rate of 1.5 m/s, which were 3.7 × 104 cells/cm2 and 1.1 × 103 CFU/cm2, respectively; (2) Proteobacteria had the highest relative abundance (RA) among all samples, and the RA in biofilm (78.85%) was higher than that in water (48%−59%); (3) when the pollutants and biofilm were partially shed, the total phosphorus (TP), permanganate index (CODMn), and dissolved organic carbon (DOC) increased by 0.011, 0.36, and 0.5 mg/L at most, respectively; and (4) dissolved oxygen (DO) was sufficient during non-full flow water transport and nitrification occurred. The highest removal rates of ammonium nitrogen (NH+4-N) and nitrous nitrogen (-N) reached 27.16% and 66.76%, respectively. At the flow rates of 2.0 m/s, the efficiency decreased to 10.47% and 41.25%, respectively, due to the shedding of biofilm.

Effects of culvert length and flow rates of raw water transport on biofilm development and water quality