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This paper examines the nitrogen isotope fractionation factors (α) associated with the volatilisation of ammonia from water under controlled conditions at two pH values (8.5 and 9.2). This experiment assumed the continuous removal of ammonia at a single purge rate of 10 mL air min -1 . The fractionation resulting from the removal of total ammonia from the water into an acid trap was named the observed isotope fractionation factor (α obs ), and it was measured as 1.019 (±0.0025) at pH 8.5 and 1.030 (±0.0025) at pH 9.2. The observed isotope fractionation factor includes the equilibrium isotope fractionation factor (α eq ) and the kinetic isotope fractionation factor (α kin ), each one mathematically derived from the experimental data. The equilibrium and kinetic isotope fractionation factors were estimated as α eq  = 1.036 (±0.0014) and α kin  = 1.050 (±0.003), respectively. Our results are compared to other previously measured and estimated fractionation factors.

The Grand River (Ontario, Canada) is impacted by wastewater treatment plants (WWTPs) that release ammonia (NH3 and NH4+) into the river. In-river microbial communities help transform this ammonia into more oxidized compounds (e.g., NO3- or N2), although the spatial distribution and relative abundance of freshwater autotrophic ammonia-oxidizing prokaryotes (AOP) are not well characterized. This study investigated freshwater N cycling within the Grand River, focusing on sediment and water columns, both inside and outside a WWTP effluent plume. The diversity, relative abundance, and nitrification activity of AOP were investigated by denaturing gradient gel electrophoresis (DGGE), quantitative real-time PCR (qPCR), and reverse transcriptase qPCR (RT-qPCR), targeting both 16S rRNA and functional genes, together with activity assays. The analysis of bacterial 16S rRNA gene fingerprints showed that the WWTP effluent strongly affected autochthonous bacterial patterns in the water column but not those associated with sediment nucleic acids. Molecular and activity data demonstrated that ammonia-oxidizing archaea (AOA) were numerically and metabolically dominant in samples taken from outside the WWTP plume, whereas ammonia-oxidizing bacteria (AOB) dominated numerically within the WWTP effluent plume. Potential nitrification rate measurements supported the dominance of AOB activity in downstream sediment. Anaerobic ammonia-oxidizing (anammox) bacteria were detected primarily in sediment nucleic acids. In-river AOA patterns were completely distinct from effluent AOA patterns. This study demonstrates the importance of combined molecular and activity-based studies for disentangling molecular signatures of wastewater effluent from autochthonous prokaryotic communities.