Two 10-day in vitro experiments were conducted to investigate the relationship between nitrogen (N) isotope discrimination (δ15N) and ammonia (NH3) emissions from sheep manure. In Exp. 1, three different manure mixtures were set up: control (C); C mixed with lignite (C + L); and grape marc (GM), with 5, 4 and 5 replications, respectively. For C, urine and faeces were collected from sheep fed a diet of 550 g lucerne hay/kg, 400 g barley grain/kg and 50 g faba bean/kg; for C + L, urine and faeces were collected from sheep fed the C diet and 100 g ground lignite added to each incubation system at the start of the experiment; for GM, urine and faeces were collected from sheep fed a diet consisting of C diet with 200 g/kg of the diet replaced with GM. In Exp. 2, three different urine-faeces mixtures were set up: 2U:1F, 1.4U:1F and 1U:1F with urine to faeces ratios of 2:1, 1.4:1 and 1:1, respectively, each with 5 replications. Lignite in C + L led to significantly lower cumulative manure-N loss by 81 and 68% in comparison with C and GM groups, respectively (P = 0.001). Cumulative emitted manure NH3-N was lower in C + L than C and GM groups by 35 and 36%, respectively (P = 0.020). Emitted manure NH3-N was higher in 2U:1F compared to 1.4U:1F and 1U:1F by 18 and 26%, respectively (P < 0.001). This confirms the relationship between manure δ15N and cumulative NH3-N loss reported by earlier studies, which may be useful for estimating NH3 losses.