ABSTRACT A lysimeter experiment was carried out to investigate the effects of applying readily available carbon (12 or 24 t sucrose ha−1) to soil on nitrogen (N) losses from cattle urine patches. The carbon (C) was readily available to microbes and was applied onto intact soil monolith lysimeters, containing stony silt loam soil, beneath either ryegrass (Lolium perenne)/white clover (Trifolium repens) pasture (RG/WC) or lucerne (Medicago sativa). Cow urine (700 kg N ha−1) was applied in early July 2017, two days after C application. The application of readily available C increased the immobilisation of N in the soil which reduced the amount of soil mineral N. The C-induced immobilisation of N reduced the ammonia oxidising bacteria population growth rate and the amount of nitrate leached by 51–89%. The addition of readily available C had no significant effect on nitrous oxide emissions. These findings were consistent under both RG/WC and lucerne.