This study examined the responses of a population of brook stickleback (Culaea inconstans) exposed to pulp mill effluent at Jackfish Bay, Lake Superior, Canada, in May 2007 and May 2011. Brook stickleback were extirpated from the effluent-receiving site, presumably due to anoxia after this period. Females at the effluent-receiving site had significantly larger gonad sizes in 2007 and 2011. In 2011, effluent-exposed female gonadal development was significantly advanced when compared with reference sites; they were the second most mature when compared among three different reference sites. Analysis of 7-ethoxyresorufin-O-deethylase activity revealed that effluent-receiving site females had greater CYP1A induction in 2007 and significantly greater CYP1A induction in 2011. Effluent-receiving site males showed significantly reduced CYP1A induction in 2007 and significantly greater induction in 2011. Chemical evaluation of sediment from the receiving environment showed elevated levels of resin acids and the polycyclic aromatic hydrocarbon, retene. Higher condition factors and more mature gonads were consistent with higher winter and spring temperatures modified by effluent or by lake vs. stream environments. Overall, effects on effluent-exposed brook stickleback were not consistent with reported effects in white sucker exposed to the same effluent in previous studies.