Tobacco is the most widespread commercial non-feed crop in the world. Tobacco production in Croatia is still a very profitable economic activity, whose production can be endangered by the pests. The tobacco flea beetle (Epitrix hirtipennis (Melsheimer, 1847)) is one of the most significant tobacco pests which, by eating the leaves, reduce the yield. The aim of the study was to determine the fauna, the intensity of attack and the damage caused by the tobacco flea beetle on tobacco F1 two line hybrids DH 17 and DH 27 in the area of Virovitica-Podravina County, and to investigate the effectiveness of botanical insecticide, organic fertilizer and chemical insecticide on most common tobacco flea beetle species in the area. In 2016, on two sites in Virovitica-Podravina County adult tobacco flea beetles were collected weekly from mid-August to early October. At the same time, to assess the damages made by flea beetles, 40 leaves of both DH 17 and DH 27 hybrids were collected weekly. The efficacy of azadirachtin (botanical insecticide), organic nitrogen fertilizer and chlorantraniliprole (chemical insecticide) on adult tobacco flea beetles collected in the same county was investigated in a laboratory experiment. The variants were applied by treating the leaves according to the IRAC No7 method, in four replicates. The test was read every 24 hours for four days. The number of dead flea beetles on insecticide and control was used to calculate the efficacy by using the Schneider-Orelli formula. All collected flea beetles were morphologically identified as species Epitrix hirtipennis, the tobacco flea beetle. The results on leaf damage showed there were significantly higher damages on DH 17 hybrid (94%) compared to DH 27 (65%). The results of insecticide efficacy showed that, although differences between the variants were not significant, the best efficacy was achieved by the botanical insecticide azadirachtin (92%). The efficacy was the same as that of a chemical insecticide chlorantraniliprole. Organic fertilizer efficiency was lower (60%), which cannot be considered satisfactory in tobacco flea beetle control. Research has shown that azadirachtin, due to favorable ecotoxicological properties, could be used as an alternative to chemical insecticides or in combination with other insecticides in tobacco flea beetle management.