Fish-eating waterbirds from the Great Lakes of North America have shown symptoms of poisoning similar to those observed in laboratory exposures of various avian species to planar halogenated hydrocarbons (PHHs). PHHs, include among others, polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) and have been implicated in some of the reproductive problems of Great Lakes waterbirds. The objectives of this study were to assess the overall potencies of PCB-containing extracts from colonial water-bird eggs taken from the Great Lakes and to compare the potencies with the location and spatial distribution of the colonies. The potencies of the extracts were assessed by their ability to induce cytochrome P450IA1-associated ethoxyresorufin O-deethylase (EROD) activity in H4IIE rat hepatoma cells as compared to the standard, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The H4IIE bioassay-derived TCDD-equivalents (TCCD-EQs) in the waterbird eggs concur with residue analyses and biological data from other studies. The greatest concentrations of TCDD-EQs were found in waterbird eggs from historically polluted, industrialized or urbanized areas in which the reproductive impairment of colonial waterbirds was most severe. However, significant concentrations of TCDD-EQs were detected at all sites tested; with a range of 49 to 415 pg TCDD-EQ/g egg, uncorrected for extraction efficiencies. The H4IIE bioassay proved to be a useful biomonitoring tool to assess the overall potency of complex PHH mixtures in environmental samples.