The James River Treatment Plant (JRTP) operated a 2 MGD Integrated Fixed Film Activated Sludge (IFAS) demonstration process from November 2007 to April 2009 to explore IFAS performance and investigate IFAS technology as an option for a full scale plant upgrade in response to stricter nutrient discharge limits in the James River Basin. During the study, nitrification kinetics for both ammonia and nitrite oxidizing bacteria and plastic biofilm carrier biomass content were monitored on a near-weekly basis comparing the IFAS media, the IFAS process mixed liquor, and mixed liquor from the full-scale activated sludge process. Carrier biomass content is variable with respect to temperature and process SRT and relates to the localization of nitrification activity in the IFAS basin. Similar to trends observed for carrier biomass content (Regmi, 2008), ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) activity also shifted from the fixed film to the suspended phase as water temperatures increased and vice versa as the temperature decreased. The data suggest that AOB activity occurs on the surface of the biofilm carriers, while NOB activity remains deeper in the biofilm. During the highest temperatures observed in the IFAS tank, AOB activity on the media contributed as little as 30% of the total nitrification activity in the basin, and after temperatures dropped below 20 °C, AOB activity in the fixed film phase made up 75% of the total activity in the IFAS basin. During the warmest period of the summer, the media still retained more than 60% of the total NOB activity, and more than 90% of the total NOB activity during the period of coldest water temperature. This trend also points out that some AOB and NOB activity remained in the mixed liquor, even during the coldest periods. The retention of nitrification activity in the mixed liquor indicates that the constant sloughing of biomass off of the carriers allowed for autotrophic activity, even during washout conditions. Carrier biomass content and nitrification rates on the IFAS media remained constant along the length of the basin, indicating that the IFAS tank is will mixed with respect to biomass growth, although there was a concentration gradient for soluble species (NH₄-N, NO₂-N, NO₃-N). In addition to the weekly nitrification rate measurements, experiments were also conducted to determine how operational inputs such as dissolved oxygen (DO) and mixing affect the nitrification rates. Mixing intensity had a clear impact on nitrification rates by increasing the velocity gradient in the bulk liquid and decreasing the mass transfer boundary layer mass transfer resistance. At higher mixing intensities, advection through the mass transfer boundary layer increased making substrate more available to the biofilm. The affect of mixing was much more profound at low DO, whereas increased mixing had less effect on nitrification rates at higher bulk liquid DO. DO also affected nitrification rates, such that as DO increased it penetrated deeper into the biofilm increasing the nitrification rate in a linear fashion until the biofilm became saturated. Another aspect of the research was modeling effective half saturation effects for AOB and NOB activity in the fixed film phase. The modeling work demonstrated that KS for AOB activity on the media was similar to accepted suspended growth KS values, while KS for NOB activity on the media was considerably higher than suspended growth KS. This trend indicates that nitrite was not as bioavailable in the biofilm and resists diffusion into the deeper part of the biofilm where NOB activity takes place. KO for both AOB and NOB activity in the biofilm was higher than typical suspended growth values because of boundary layer and biofilm diffusion resistances. In addition, the presence of readily degradable organics did not significantly affect nitrification rates on the media, but did reduce nitrification rates in the mixed liquor. That, combined with low chemical oxygen demand (COD) uptake rates indicates that little heterotrophic activity is occurring on the media.