Stormwater biofilter response to high nitrogen loading under transient flow conditions: Ammonium and nitrate fates, and nitrous oxide emissions.

Nitrogen (N) in urban runoff is often treated with green infrastructure including biofilters. However, N fates across biofilters are insufficiently understood because prior studies emphasize low N loading under laboratory conditions, or use "steady-state" flow regimes over short time scales. Here, we tested field scale biofilter N fates during simulated storms delivering realistic transient flows with high N loading. Biofilter outflow ammonium (NH4+-N) was 60.7 to 92.3% lower than that of the inflow. Yet the characteristic times for nitrification (days to weeks) and denitrification (days) relative to N residence times (7 to 30 h) suggested low N transformation across the biofilters. Still, across 7 successive storms, total outflow nitrate (NO3--N) greatly exceeded (3100 to 3900%) inflow nitrate, a result only explainable by biofilter soil N nitrification occurring between storms. Archaeal, and bacterial amoA gene copies (2.1 × 105 to 1.2 × 106 gc g soil-1), nitrifier presence by16S rRNA gene sequencing, and outflow δ18O-NO3- values (-3.0 to 17.1 ‰) reinforced that nitrification was occurring. A ratio of δ18O-NO3- to δ15N-NO3- of 1.83 for soil eluates indicated additional processes: N assimilation, and N mineralization. Denitrification potential was suggested by enzyme activities and soil denitrifying gene copies (nirK + nirS: 3.0 × 106 to 1.8 × 107; nosZ: 5.0 × 105 to 2.2 × 106 gc g soil-1). However, nitrous oxide (N2O-N) emissions (13.5 to 84.3 µg N m - 2 h - 1) and N2O export (0.014 g N) were low, and soil nitrification enzyme activities (0.45 to 1.63 mg N kg soil-1day-1) exceeded those for denitrification (0.17 to 0.49 mg N kg soil-1 day-1). Taken together, chemical, bacterial, and isotopic metrics evidenced that storm inflow NH4+sorbs and, along with mineralized soil N, nitrifies during biofilter dry-down; little denitrification and associated N2O emissions ensue, and thus subsequent storms export copious NO3--N. As such, pulsed pass-through biofilters require redesign to promote plant assimilation and/or denitrification of mineralized and nitrified N, to minimize NO3--N generation and export.

Highly variable removal of pathogens, antibiotic resistance genes, conventional fecal indicators and human-associated fecal source markers in a pilot-scale stormwater biofilter operated under realistic stormflow conditions.

Green stormwater infrastructure systems, such as biofilters, provide many water quality and other environmental benefits, but their ability to remove human pathogens and antibiotic resistance genes (ARGs) from stormwater runoff is not well documented. In this study, a field scale biofilter in Southern California (USA) was simultaneously evaluated for the breakthrough of a conservative tracer (bromide), conventional fecal indicators, bacterial and viral human-associated fecal source markers (HF183, crAssphage, and PMMoV), ARGs, and bacterial and viral pathogens. When challenged with a 50:50 mixture of untreated sewage and stormwater (to mimic highly contaminated storm flow) the biofilter significantly removed (p < 0.05) 14 of 17 microbial markers and ARGsin descending order of concentration reduction: ermB (2.5 log(base 10) reduction) > Salmonella (2.3) > adenovirus (1.9) > coliphage (1.5) > crAssphage (1.2) > E. coli (1.0) ∼ 16S rRNA genes (1.0) ∼ fecal coliform (1.0) ∼ intl1 (1.0) > Enterococcus (0.9) ∼ MRSA (0.9) ∼ sul1 (0.9) > PMMoV (0.7) > Entero1A (0.5). No significant removal was observed for GenBac3, Campylobacter, and HF183. From the bromide data, we infer that 0.5 log-units of attenuation can be attributed to the dilution of incoming stormwater with water stored in the biofilter; removal above this threshold is presumably associated with non-conservative processes, such as physicochemical filtration, die-off, and predation. Our study documents high variability (>100-fold) in the removal of different microbial contaminants and ARGs by a field-scale stormwater biofilter operated under transient flow and raises further questions about the utility of human-associated fecal source markers as surrogates for pathogen removal.