Abstract: Predicting impact of oil spills on aquatic life requires a better understanding of effects on aquatic organisms, both for single hydrocarbons and for their interactions. In this study, the individual and combined effects of petroleum hydrocarbons phenanthrene (Phen) and dibenzothiophene (DBT) were assessed on the reproductive behavior of the freshwater amphipod Hyalella azteca. Following a 24-h exposure to single PAHs, or an equimolar mixture of Phen-DBT, mate-guarding behavior was assessed at the end of the exposure and during a subsequent 10 min behavioral observation period with the animals in clean water. The endpoints of the study during the behavior observation period were—time taken to initiate mate-guarding (TIMG), and proportion of time spent in amplexus (PTA). The study demonstrated that the exposure to Phen and DBT reduced the incidence of mate-guarding during the actual exposure period, but not during the observation period. However, whether or not pairs were involved in mate-guarding at the end of the exposure period did affect both TIMG and PTA during the observation period. Thus, the effects of Phen and DBT on amplexus status at the end of the exposure period indirectly affected TIMG and PTA during the observation period. The interaction between Phen and DBT with respect to their effects on mate-guarding varied among the mate-guarding measures. For the amplexus status at the end of the exposure period and for the effect on TIMG, the interaction did not deviate statistically from an additive effect. For PTA, the overall interaction was a synergistic one. This study\'s findings point out that assessments of hydrocarbon toxicity need to take into account that subtle reproductive behaviors (that may play an important role in population persistence) may be negatively affected. The results also show that the general assumption of additive effects among different PAHs may be an oversimplification.