The development and proliferation of antibiotic resistance in pathogenic, commensal, and environmental microorganisms is a major public health concern. The extent to which human activities contribute to the maintenance of environmental reservoirs of antibiotic resistance is poorly understood. In the current study, wastewater treatment plants (WWTPs) were investigated as possible sources of tetracycline resistance via qualitative PCR and quantitative PCR (qPCR). Various WWTPs and two freshwater lakes were surveyed for the presence of an array of 10 tetracycline resistance determinants (tet(R)): tet(A)-(E), tet(G), tet(M), tet(O), tet(Q), tet(S). All WWTP samples contained more different types of tet(R) genes, as compared to the lake water samples. Gene copy numbers of tet(G) and tet(Q) in these samples were quantified via qPCR and normalized to both the volume of original sample and to the amount of DNA extracted per sample (a proxy for bacterial abundance). Concentrations of tet(Q) were found to be highest in wastewater influent while tet(G) concentrations were highest in activated sludge. Investigation of the effects of UV disinfection on wastewater effluent showed no reduction in the number of detectable tet(R) gene types.