Estuaries of the northern Gulf of Mexico are dynamic environments, with fluctuations in salinity and dissolved oxygen, including areas of seasonal hypoxia. Fish that reside and reproduce in these estuaries, including sheepshead minnow (Cyprinodon variegatus; SHM), were at significant risk of oil exposure following the Deepwater Horizon oil spill. It is poorly understood how differences in environmental conditions during oil exposure impact its toxicity. The present study investigated the effects of crude oil high-energy water accommodated fraction (HEWAF) on SHM reproduction in three environmental scenarios (normoxic, hypoxic, and hypoxic with low salinity) to determine if differences in salinity (brackish vs low salinity) and dissolved oxygen (normoxia vs hypoxia) could exacerbate the effects of HEWAF-derived polycyclic aromatic hydrocarbons (PAHs). We observed that HEWAF exposure significantly increased liver somatic index of SHM compared to control, but this effect was not exacerbated by hypoxia or low salinity. HEWAF exposure also significantly decreased egg production and egg fertilization rate, but only in the hypoxic and hypoxic with low salinity scenarios. A significant correlation existed between body burdens of PAHs and reproductive endpoints, providing substantial evidence that oil exposure reduced reproductive capacity in SHM, across a range of environmental conditions. These data suggest that oil spill risk assessments that fail to consider other environmental stressors (i.e. hypoxia and salinity) may be underestimating risk.Mississippi, Alabama, and Florida were all impacted by oil slicks that washed ashore (Nixon et al., 2016). In late April 2010, in an effort to limit the effects of oil on coastal habitats, the state of Louisiana initiated a massive diversion of freshwater from the Mississippi River into Barataria Bay and Breton Sound (Bianchi et al., 2011;Martínez et al., 2012). These freshwater diversions significantly reduced salinities in the estuaries of the northern GOM (Bianchi et al., 2011;Powers et al., 2017), but it is still undetermined if this effort was effective in reducing oiling to the Louisiana coast (Martínez et al., 2012).Estuaries of the northern GOM are dynamic environments, with constant fluctuations in salinity and dissolved oxygen. Freshwater inflow from the Mississippi River causes a saline gradient in the estuaries that contributes to water stratification, usually during summer months (USEPA, 1999). The combination of water stratification, excess