From a discarded heavy-equipment tire (ca. 57 liter) at an industrial construction site, we collected 655 (86.0%) Orthopodomyia signifera, 23 (3.0%) Toxorhynchites rutilus septentrionalis, 17 (2.2%) Aedes japonicus japonicus, and 67 (8.8%) Culex pipiens pipiens. Although larvae of Aedes albopictus and Aedes triseriatus were not collected from this container, both species were prevalent as host-seeking adults and readily collected as larvae from other containers at this site. Laboratory trials to test the survival of prey (Ae. albopictus, Cx. p. pipiens, or Or. signifera) in the presence of Tx. rut. septentrionalis showed that survival of prey larvae differed among species. Multiple comparisons revealed that Ae. albopictus had the lowest and Or. signifera the highest survival in the presence of Tx. rut. septentrionalis. Survival of Or. signifera and Cx. p. pipiens was not significantly different from one another, but both were different from Ae. albopictus. Further testing is warranted to test other factors responsible for differences in the interspecific relationship between Or. signifera and other species in tree hole communities.