Fertiliser and manure application are important sources of nitrous oxide (N2O) emissions from agricultural soils. The current default IPCC emission factor of 1.0% is independent of the type of fertiliser and manure, and application time, method and rate. However, in the IPCC Tiered system it is possible to use more specific emission factors that better reflect the actual fertiliser and manure management in a given country or region. The first and primary aim of this study was to determine whether the combination of cattle slurry injection with fertiliser application, which is common practice in intensively managed grasslands in the Netherlands and neighbouring countries, warrants an adjusted emission factor. A second aim was to evaluate whether alternative emission factors, based on N uptake and N surplus, respectively, give more insight in the N2O emission rates of various fertilisation strategies. In a 2-year field experiment on sandy soil in the Netherlands we measured the annual N2O emission from grasslands receiving repeated simultaneous applications of fertiliser and cattle slurry. The N2O fluxes and N uptake by grass were measured from plots receiving calcium ammonium nitrate (CAN) at four application rates, either with or without additional application of liquid cattle slurry, applied through shallow soil injection. The average emission factor for fertiliser-only treatments was 0.15%. The annual N2O emissions were similar for treatments receiving only fertiliser or only cattle slurry. In the first experimental year, application of cattle slurry increased the emission factor for fertiliser to 0.35%, but the second year showed no effect of cattle slurry on the emission from fertiliser. With regard to the first objective, we conclude that these results do not conclusively justify an adjusted emission factor for combined application of fertiliser and cattle slurry. To minimise risks however, it is sensible to avoid simultaneous application of fertiliser and cattle slurry. The N2O emission factor expressed as percentage of kg N uptake by grass was consistently higher after combined application of fertiliser and cattle slurry (0.29%), compared to fertiliser-only (0.17%). With regard to the second objective we conclude that an emission factor based on N uptake expresses the relatively inefficient N supply of cattle slurry to crop growth better than the traditional emission factor based on N application.