In constructed wetlands (CWs) with horizontal sub-surface flow, nutrient removal, especially phosphorus, is limited because the root biomass fills the pore spaces of the substrate (usually gravel), directing wastewater flow to deeper wetland media; plants are not regularly harvested; the litter formed by decomposing vegetation remains on the surface of the substrate and thus does not interact with the wastewater; and the substrate media often used provide only limited adsorption. Effective nutrient removal including rootzone oxidation, adsorption and plant uptake therefore requires sufficient interaction of wastewater with the treatment media. We assessed the feasibility of biological nutrient removal from wastewater using substrate-free CWs with horizontal flow, planted with two tropical macrophytes namely, Cyperus papyrus and Miscanthidium violaceum. The objectives were to evaluate the system treatment efficiency under semi-natural conditions, and to assess microbial and plant biomass contributions to nutrient removal in the CWs. Results showed high removal efficiencies for biochemical oxygen demand, ammonium-nitrogen (NH 4 -N) and phosphorus (P) fractions in papyrus-based CWs (68.6-86.5%) compared to Miscanthidium (46.7-61.1%) and unplanted controls (31.6-54.3%). Ammonium oxidizing bacteria in CW root-mats (10 8 -10 9 cells/gram dry weight) and residual nitrite and nitrate concentrations in the water phase indicated active system nitrification. Papyrus showed higher biomass production and nutrient uptake, contributing 28.5% and 11.2%, respectively, of the total N and P removed by the system compared to 15% N and 9.3% P removed by Miscanthidium plants. Compared to literature values, nitrification, plant uptake and the overall system treatment efficiency were high, indicating a high potential of this system for biological nutrient removal from wastewaters in the tropics.
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