Entomopathogenic nematodes (EPNs) in the genera Steinernema and Heterorhabditis (Rhabditida: Steinernematidae and Heterorhabditidae, respectively) represent a most promising group of pathogens that are lethal to many soil-inhabiting insects. Ukraine has Lagged behind of Eastern European countries in apply studies of entomopathogenic nematodes. Recently, Heterorhabditis bacteriophora were extracted from soils in the Central Forest Steppe areas of Ukraine using Galleria mellonella traps and identified with PCR. Infection were carried out in Laboratory to determine the host range, specificity and virulence of H. bacteriophora ukr. isol. It was shown that this nematode infected a wide range of insect species. Of the 14 tested insects which belong to four orders , all were susceptible to nematode species with mortality ranges from 12 to 96%. Infectivity did differ within each host. The most favorable hosts were Lepidopterans and coleopterans. The virulence of H. bacteriophora was measured by one-on-one bioassay. H. bacteriophora caused 60% mortality at 5 infective juveniles per Galleria mellonella larvae. LD50 values were 3 +/- 1. Nematodes reproduced and completed their cycle with IJ production in all four orders. H. bacteriophora ukr. isoL.. developed preferably in immature insect stage. Percentage of emergence form cadavers that produced progeny were higher for Lepidoptera and Coleopteran species. Progeny production were not related to the biomass of the cadavers. Because of wide range of insect species Heterhorobditis bacteriophora ukr. isol. has potential for control economically important pests in Ukraine. Further studies should be focused at ecological characterization of isolated H. bacteriophora and carrying out survey of entomopathogenic nematodes.