Levofloxacin is a broad-spectrum antibacterial drug in the fluoroquinolone class that is classified as category C during pregnancy. This study aimed to determine the effects of levofloxacin administration on embryos at different developmental stages. It examined 420 fertile chicken eggs, divided into two main groups-levofloxacin treatment on days 7 and 14-each containing seven subgroups (n = 30 each): negative control (no treatment), positive control (treated with saline), and five levofloxacin doses (5000, 2500, 1250, 625, and 312.5 µg/kg). On day 21, all eggs were hatched, the live-dead ratio was determined, and liver and kidney tissues were collected for histopathological analysis. Administering levofloxacin to embryos at 7 and 14 days of incubation had no lethal effect at doses up to 5000 µg/kg (p>0.05). Histopathological evaluations revealed no significant or clinically meaningful histopathological toxicity was observed of levofloxacin on liver and kidney development (p>0.05). In conclusion, when administered directly, levofloxacin does not have adverse effects on embryos at different developmental stages.