The activated sludge process in Wastewater Treatment Plant (WWTPs) relies on the activities of microbes to reduce the organic and inorganic matter and produce effluent that is safe to discharge into receiving waters. This research examined the effects of non-steroidal antiinflammatory drug (NSAID) ibuprofen and the antibiotic tetracycline on the microbial population in activated sludge from the Humber WWTP. The current investigation was designated to observe the impact of these contaminants, at low (environmentally relative concentrations) as well as extremely high concentrations of tetracycline and ibuprofen. Using 16S and 18S rRNA gene primer sets, and qPCR the abundance of each population was monitored as well as the relative abundance of two populations under the various conditions. It was found that current environmental concentrations of ibuprofen stimulated protozoan growth but higher concentrations reduced their numbers especially in the presence of tetracycline. Finally using DGGE, the identity for some of the more abundant protozoa were identified and it was noted that high ibuprofen and tetracycline concentrations favored the abundance of some genera.