Considering the increasing antibiotics resistance, there has been a propensity to replace them with antiseptics when it comes to wound management and treatment. Nevertheless, in recent years, there have been reports regarding resistance to antiseptics by some bacterial strains. There is also concern about the environmental impact of these substances. The aim of this study was to compare the antimicrobial efficacy of antiseptics and eucalyptus essential oils on bacterial strains from horse’s wounds. We used twelve Escherichia coli, eight Staphylococcus aureus, two Staphylococcus pseudintermedius, one Staphylococcus vitulinus and one Staphylococcus saprophyticus strains from equine wounds. The effect of Eucalyptus radiata essential oil, Eucalyptus globulus essential oil, povidone-iodine and chlorhexidine gluconate against the isolated strains was evaluated applying the Kirby-Baüer method. Regarding the Escherichia coli strains, E. radiata and the mixture of E. radiata and E. globulus had a better inhibitory effect than antiseptics. E. globulus had a better effect against most Staphylococcus spp. compared to E. radiata. For both Gram-negative and Gram-positive strains tested, chlorhexidine gluconate had a better inhibitory effect than povidone-iodine. The antibacterial efficacy of essential oils highlights their potential to substitute or complement the use of antiseptics and so reduce resistance to antiseptics.