Third-stage larvae of the nematode Serpinema trispinosum (Leidy, 1852) were collected from the midgut of four of five species of adult damselflies (Zygoptera) from a non-irrigated restored semipermanent wetland located in Stillwater, Oklahoma, USA. Of the four infected damselfly species, prevalence and mean abundance was highest for the southern spreadwing, Lestes disjunctus australis Walker (10%, 0.2 ± 0.8) and lowest for the familiar bluet, Enallagma civile (Hagen) (2.5%, 0.04 ± 0.3); whereas mean intensities were lowest for the citrine forktail, Ischnura hastata (Say) (1.5 ± 0.5) and the eastern forktail, Ischnura verticalis (Say) (1.0 ± 0). This is the first record of larvae of S. trispinosum from damselflies. Serpinema trispinosum adults have been reported from 18 species of North and Central American freshwater turtles, whereas microcrustaceans such as copepods serve as intermediate hosts and snails, fish and amphibians serve as paratenic hosts in this nematode's life cycle. However, dietary studies of the 18 species of freshwater turtles reported as definitive hosts for S. trispinosum indicate that aquatic insects including damselflies are more commonly reported in turtle diets than are fish or amphibians. Additionally, unlike snails and amphibians, larval damselflies predominantly feed on microcrustaceans, and our observation of S. trispinosum infecting damselflies may reflect the importance of these insects as paratenic hosts. In the present study, we provide new host information and measurements for third-stage larvae of S. trispinosum from damselfly hosts along with measurements for adult male and female S. trispinosum from turtle hosts from Oklahoma, USA.