There is increased focus on nitrogen (N)-containing dissolved organic matter (DOM) as a nutrient source supporting eutrophication in N-sensitive estuarine ecosystems. This is particularly relevant in watersheds undergoing urban and agricultural development, leading to increased dissolved organic N (DON) loading. To understand how this shift in N-loading influences estuarine phytoplankton production, nutrient addition bioassays were conducted in the N-limited Neuse River Estuary, North Carolina from 2014 to 2015. Additions included N-rich DOM sources characteristic of urban and agricultural development, including chicken and turkey litter leachate, wastewater treatment facility effluent, and concentrated river DOM (used as a reference). Each DOM addition was coupled with an inorganic nutrient treatment to account for inorganic nutrient concentrations (NO2/3, NH4, PO4) in each respective DOM addition. Repeated measures analysis of variance (RM-ANOVA) showed that chicken litter leachate stimulated phytoplankton growth greater than its coupled inorganic nutrient treatment. Wastewater treatment facility effluent, turkey litter leachate, and concentrated river DOM did not stimulate phytoplankton growth greater than their respective inorganic nutrient controls. DOM fluorescence (EEM-PARAFAC) indicated the chicken litter contained a biologically reactive fluorescent DOM component, identified as the nonhumic, biologically labile, "N-peak", which may be responsible for stimulating the observed phytoplankton growth in the chicken litter leachate treatments.