Bioassays for baseline toxicity, which measure toxicants’ non-specific effects, have been shown in previous studies to effectively correlate with the increased presence of pharmaceuticals, personal care products, endocrine-disrupting compounds, and other synthetic organics in treated sewage effluent. This study investigated how the baseline toxicity of anthropogenic compounds-spiked wastewater changed during the treatment of biofiltration and ozone oxidation, as measured by the bioluminescence inhibition of the Vibrio fischeri bacterium. The water quality parameters of dissolved organic carbon, seven common anions, and fluorescence spectroscopy were used to corroborate and collate with the toxicity results. Water quality was evaluated on two bench-scale soil filtration columns, which were configured for pre-ozonation and post-ozonation. Both systems’ soil aerobically removed similar amounts of dissolved organic carbon, and the reduction ranged between 57.7% and 62.1% for the post-ozonation and pre-ozonation systems, respectively. Biological removal of DOC, protein-like, humic-like, and soluble microbial product-like material was highest in the first 28.5 cm of each 114 cm-long system. While bioluminescence inhibition showed that ozonation was effective at lowering baseline toxicity, this study’s bioassay procedure was a very poor indicator of soil filtration treatment; both system’s effluents were significantly more toxic than their non-ozonated influents.