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Urbanization growth has intensified the challenge of managing and treating increasing amounts of municipal solid waste (MSW). Landfills are commonly utilized for MSW disposal because of their low construction and operation costs. However, this practice produces huge volumes of landfill leachate, a highly polluting liquid rich in ammoniacal nitrogen (NH3-N), organic compounds, and various heavy metals, making it difficult to treat in conventional municipal wastewater treatment plants (WWTPs). In recent years, research has shown that microbial biofilms, developed on carriers of different materials and called “moving bed biofilm reactors” (MBBRs), may offer promising solutions for bioremediation. This study explored the biofilm development and the nitrification process of moving bed biofilms (MBBs) obtained from high ammonia-selected microbial communities. Using crystal violet staining and confocal laser-scanning microscopy, we followed the biofilm formation stages correlating nitrogen removal to metagenomic analyses. Our results indicate that MBBs unveiled a 10-fold more enhanced nitrification rate than the dispersed microbial community present in the native sludge of the Porto Sant’Elpidio (Italy) WWTP. Four bacterial families, Chitinophagaceae, Comamonadaceae, Sphingomonadaceae, and Nitrosomonadaceae, accumulate in structured biofilms and significantly contribute to the high ammonium removal rate of 80% in 24 h as estimated in leachate-containing wastewaters.

The increasing amounts of municipal solid waste and their management in landfills caused an increase in the production of leachate, a liquid formed by the percolation of rainwater through the waste. Leachate creates serious problems to municipal wastewater treatment plants; indeed, its high levels of ammonia are toxic for bacterial cells and drastically reduce the biological removal of nitrogen by activated sludge. In the present work, we studied, using a metagenomic approach based on next-generation sequencing (NGS), the microbial composition of sludge in the municipal wastewater treatment plant of Porto Sant’Elpidio (Italy). Through activated sludge enrichment experiments based on the Repetitive Re-Inoculum Assay, we were able to select and identify a minimal bacterial community capable of degrading high concentrations of ammonium (NH4+-N ≅ 350 mg/L) present in a leachate-based medium. The analysis of NGS data suggests that seven families of bacteria (Alcaligenaceae, Nitrosomonadaceae, Caulobacteraceae, Xanthomonadaceae, Rhodanobacteraceae, Comamonadaceae and Chitinophagaceae) are mainly responsible for ammonia oxidation. Furthermore, we isolated from the enriched sludge three genera (Klebsiella sp., Castellaniella sp. and Acinetobacter sp.) capable of heterotrophic nitrification coupled with aerobic denitrification. These bacteria released a trace amount of both nitrite and nitrate possibly transforming ammonia into gaseous nitrogen. Our findings represent the starting point to produce an optimized microorganisms’s mixture for the biological removal of ammonia contained in leachate.