The potential of dietary nitrate to reduce enteric methane, apart from it being a source of rumen-degradable nitrogen, has stimulated further research into its use. However, not much has been reported on its suitability in feed treatment. The objective of this study was to determine the effects of urea or nitrate and two methods of non-protein nitrogen (NPN) application (anaerobic pre-treatment versus direct supplementation) to Eragrostis curvula hay, on its in vitro fermentation. An iso-nitrogenous level of NPN (7 g nitrogen/kg feed dry matter (DM)) from either urea or calcium nitrate was used to pre-treat hay by subjecting it to 30 days’ anaerobic storage in airtight bottles, or by direct supplementation; each diet having three replicates. Hay samples were dried, milled, and evaluated for in vitro organic matter digestibility (IVOMD), and in vitro gas and methane production, while rumen fluid was analysed for pH, ammonia nitrogen and volatile fatty acids (VFAs). Compared with the untreated hay (control), under both methods of application NPN generally did not increase 48-hour gas production, but did increase the IVOMD of E. curvula hay. However, pre-treatment of E. curvula hay improved IVOMD more than supplementation. Urea inclusion enhanced digestibility more than nitrate, and the urea pre-treatment was more effective than supplementation. In contrast, nitrate inclusion significantly reduced methane production compared with urea and the control, and supplementation of nitrate was more effective in reducing methane than pre-treatment with nitrate. Pre-treatment with nitrate as supplementation increased digestibility and reduced enteric methane emission, indicating the potential of using nitrate as a hydrolytic agent in feed treatment.