Nitrogen(N) loss from fertilization in agricultural fields has an unavoidable negative impact on the environment and a better understanding of the major pathways can assist in developing the best management practices. The aim of this study was to evaluate the fate of N fertilizers applied to acidic red soil(Ferralic Cambisol) after 19 years of mineral(synthetic) and manure fertilizer treatments under a cropping system with wheat-maize rotations. Five field treatments were examined: control(CK), chemical nitrogen and potash fertilizer(NK), chemical nitrogen and phosphorus fertilizer(NP), chemical nitrogen, phosphorus and potash fertilizer(NPK) and the NPK with manure(NPKM, 70% N from manure). Based on the soil total N storage change in 0–100 cm depth, ammonia(NH_3) volatilization, nitrous oxide(N_2O) emission, N plant uptake, and the potential N leaching loss were estimated using a mass balance approach. In contrast to the NPKM, all mineral fertilizer treatments(NK, NP and NPK) showed increased nitrate(NO_3~–) concentration with increasing soil depth, indicating higher leaching potential. However, total NH_3 volatilization loss was much higher in the NPKM(19.7%) than other mineral fertilizer treatments(≤4.2%). The N_2O emissions were generally low(0.2–0.9%, the highest from the NPKM). Total gaseous loss accounted for 1.7, 3.3, 5.1, and 21.9% for NK, NP, NPK, and NPKM treatments, respectively. Estimated N leaching loss from the NPKM was only about 5% of the losses from mineral fertilizer treatments. All data demonstrated that manure incorporation improved soil productivity, increased yield, and reduced potential leaching, but with significantly higher NH_3 volatilization, which could be reduced by improving the application method. This study confirms that manure incorporationis an essential strategy in N fertilization management in upland red soil cropping system. Nitrogen (N) loss from fertilization in agricultural fields has an unavoidable negative impact on the environment and a better understanding of the major pathways assist in developing the best management practices. The aim of this study was to evaluate the fate of N fertilizers applied to acidic red soil (Ferralic Cambisol) after 19 years of mineral (synthetic) and manure fertilizer treatments under a cropping system with wheat-maize rotations. Five field treatments were examined: control (CK), chemical nitrogen and potash fertilizer (NK) nitrogen and phosphorus fertilizer (NP), chemical nitrogen, phosphorus and potash fertilizer (NPK) and the NPK with manure (NPKM, 70% N from manure) NH 3) volatilization, nitrous oxide (N 2 O) emission, N plant uptake, and the potential N leaching loss were estimated using a mass balance approach. In contrast to the NPKM, all mineral fertilizer treatments (NK, NP and NPK) showed However, total NH_3 volatilization loss was much higher in the NPKM (19.7%) than other mineral fertilizer treatments (≤4.2%). The N_2O emissions were generally low (0.2-0.9%, the highest from the NPKM). Total Whole Loss accounted for 1.7, 3.3, 5.1, and 21.9% for NK, NP, NPK, and NPKM treatments, respectively. Estimated N leaching loss from the NPKM was only about 5% of the losses from mineral fertilizer treatments. All data for that manure incorporation improved soil productivity, increased yield, and reduced potential leaching, but with significantly higher NH 3 volatilization, which could be reduced by improving the application method. manure incorporationis an essential strategy in N fertilization management in upland red soil cropping system.