Characteristics of biofilm layer in a bio-doubling reactor and their impact on aerobic denitrifying bacteria enrichment.

Microbial loss significantly affects wastewater treatment efficiency. This study simulated the inoculation area of a self-developed biological doubling reactor (BDR) to evaluate the retention efficiency of seven different fillers for aerobic denitrifying bacteria. Over 90 days of continuous operation, the porous filler R3 demonstrated excellent performance, with OD values consistently exceeding 1.0 and minimal fluctuation. On day 90, the seed liquid amplified with R3 achieved removal efficiencies of 100% for ammonia nitrogen, 97.75% for total nitrogen, and 96.4% for chemical oxygen demand, outperforming other fillers. Scanning electron microscopy and microscopic analysis revealed that R3's large large specific surface area and volume formed a unique meshed biofilm structure, enhancing oxygen and nutrient transport while minimizing detachment. This promoted effective enrichment and retention of aerobic denitrifying bacteria. Microbial diversity analysis confirmed that Acinetobacter, a key genus involved in aerobic denitrification, dominated the network biofilm on R3, accounting for an average of 35.63%. while granular fillers, due to oxygen limitation, promoted the growth of anaerobic ammonium-oxidizing Alcaligenes. The use of BDR-enhanced MBBR for treating synthetic wastewater resulted in a 29.6% increase in TN removal efficiency, with stable system operation. The use of porous fillers with a high specific volume supports stable biofilm formation and consistent seed liquid output, providing a viable solution to microbial loss in wastewater treatment processes.