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Abstract Background Essential oils and antimicrobial peptides are two well-known safe and natural products that have been considered as alternatives to antibiotics. In the present study, the antibacterial activity of four plant essential oils and one lactoferrin-derived peptide was investigated. Results The chemical profile of each essential oil was determined by GC and GC–MS. Antimicrobial activity was shown against seven clinically isolated veterinary pathogens. MIC and MBC assessment of the essential oils and cLFchimera exhibited different antibacterial properties (MIC from a range of 62.5 to 500 µg/mL and 3.5 to 39.0 µg/mL for essential oils and cLFchimera, respectively). Compared to the essential oils, cLFchimera showed more significant antibacterial activity. Among the essential oils, Vitex agnus-castus and Salvia officinalis showed relatively better antibacterial activity. Conclusions The in vitro results reported here suggested that, for animals suffering from these pathogens, cLFchimera and the essential oils particularly Vitex agnus-castus could be considered as potential antimicrobial agents.