Bacillus thuringiensis Berliner toxins are highly specific in terms of insect pest targeted. For example, Cry3Bb is active against Coleoptera. As a consequence, in the framework of using genetically modified plants expressing different Bt toxins in sustainable agriculture, there is a growing interest about the impact of these crops on the environment and on non-target organisms. The effects of genetically modified Bt-tomato plants expressing the toxin Cry3Bb on the aphid Macrosiphum euphorbiae (Thomas) and its natural enemies, the generalist predator Macrolophus caliginosus Wagner and the endoparasitoid wasp Aphidius ervi Haliday, were assessed under laboratory conditions. No significant differences between performance of M. euphorbiae were observed on genetically modified tomato plants (line UC82Bt) with respect to their near-isogenic control line (line UC82). Immunoassays did not detect Cry3Bb protein in M. euphorbiae developing on Bt-expressing tomato plants. Similarly, no significant differences were found on the longevity and prey consumption of M. caliginosus when fed aphids reared on UC82Bt or on UC82. Moreover, the genetic modification did not affect the attractiveness of uninfested tomato plants toward A. ervi. It is therefore concluded that Cry3Bb-expressing tomato plants did not show any acute adverse effects on the biological parameters of the non-target herbivore M. euphorbiae or its natural enemies, M. caliginosus and A. ervi, under laboratory conditions.