Four entomopathogenic nematode species (Steinernema feltiae, S. carpocapsae, Heterorhabditis bacteriophora, and H. megidis) were tested in a laboratory bioassay for the efficacy of these pathogens in controlling the larvae and adults of the Colorado potato beetle, Leptinotarsa decemlineata. The main aim of the study was to develop an efficient sustainable control method against the pest. With this we could develop a strategy of potato production with the intention of diminishing or even preventing the appearance of pest resistance to insecticides. The activity of these biological agents was assessed at three different temperatures (15, 20, and 25 °C) and three concentrations (200, 1000, and 2000 infective juveniles per individual). Mortality of three stages (young and old larvae and adults) was determined 2, 4, and 7 days after treatment. At 15 °C entomopathogenic nematodes showed the lowest efficacy against all insect stages. No significant differences in efficacy was determined at 20 and 25 °C as all nematodes caused prompt death of all stages. At all temperatures young larvae were most susceptible. However, when controlling overwintered adults for the purpose of preventing the mass appearance of Colorado potato beetle, we recommend an application of higher concentrations of S. feltiae suspension.