Gastrointestinal nematodes were recovered from thirty four Gracilinanus agilis from forty four collected in Pantanal, Mato Grosso do Sul State, Brazil. Two hundred seventy four spirurids were recovered from the esophagus, stomach and intestines, comprising three species from three different genera. These were identified as Pterygodermatites (Paucipectines) jägerskiöldi, Spirura guianensis and Physaloptera herthameyerae n. sp. is first described. This is the first record of nematodes of the genera Physaloptera and Spirura in hosts of the genus Gracilinanus. The high prevalence of spirurids in 72.3% of the G. agilis collected probably is influenced by the arboreal and diet behaviors.