Stored manures are often considered to be source of nitrous oxide (N2O). A study was done to identify manure characteristics and handling stages related to N2O production. In two laboratory experiments we investigated the production of N2O from several animal manures collected at different stages of manure handling or locations in stored-manure systems. Denitrifying enzyme activity, denitrification and CO2 production rates and chemical and fibre characteristics of manure samples were also determined. Most samples had low rates of N2O production. Exceptions were open piles of fresh beef manure and the moist surface layer near the base of an open pile of poultry manure. Production of N2O was most highly correlated with nitrite plus nitrate (NO2- + NO3-) content of manure regardless of whether results were expressed on a wet or a dry weight basis. Denitrification was the most probable source of N2O because N2O production with acetylene and denitrifying enzyme activities was higher than N2O production without acetylene. Stored manure is potentially an important source of N2O emissions, particularly when storage conditions are conducive to formation of (NO2- + NO3-).