Background and aimsThe intake of selenium, an essential element for animals and humans, in ruminants is largely determined by selenium concentration in ingested forages, which take up selenium mainly from soil. Ruminant excreta is a common source of organic fertilizer, which provides both nutrients and organic matter. This study aims to unentangle the unclear effect of applying different types of ruminant excreta in soils of different organic matter contents on selenium uptake by forage.MethodsPerennial ryegrass (Lolium perenne) was grown in soils of different organic matter contents. Urine and/or feces collected from sheep fed with organic or inorganic mineral supplements, including selenium, were applied to the soils. The selenium in the collected samples were analyzed using ICP-MS. The associated biogeochemical reactions were scrutinized by wet chemistry.ResultsThe application of urine and/or feces resulted in either the same or lower selenium concentrations in perennial ryegrass. The excreta type did not affect total selenium accumulation in grass grown in low organic matter soil, whereas in high organic matter soil, feces resulted in significantly lower total selenium accumulation than urine, which was attributed to a possible interaction of selenium sorption in soil and microbial reduction of Se.ConclusionThis one-time excreta application did not increase, but further decrease in some treatments, selenium concentration and accumulation in the perennial ryegrass. Consequently, to increase ruminant selenium intake, supplementing selenium directly to animals is more recommended than applying animal manure to soil, which might drive selenium reduction and decrease selenium uptake by grass.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11104-023-05898-8.