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Author Contributions: All authors performed the experiments and contributed to data acquisition and analysis. Conceptualization: G.O.-D., A.B. and A.M.-T.; Methodology: A.M.-T., J.J.A. and T.A.; Validation: G.O.-D., A.B., M.A.A.-C., T.A. and S.P.; Writing, review and editing: M.A.A.-C., A.B., G.O.-D., S.P., T.A. and A.M.-T.; supervision, A.B. and G.O.-D. All authors have read and agreed to the published version of the manuscript., Background: The increase of multi-resistant bacteria, especially Staphylococcus spp. and Enterobacteriaceae, constitutes a challenge in veterinary medicine. The rapid growth of resistance is outpacing antibiotic discovery. Innovative strategies are needed, including the use of natural products like Allium species (Allium sativum L. and Allium cepa L.), which have been used empirically for centuries to treat infectious diseases in humans and farm and aquaculture animals due to their antibacterial properties. Methods: This study aimed to evaluate the in vitro activity of two Alliumderived compounds, propyl propane thiosulfinate (PTS) and propyl propane thiosulfonate (PTSO), against multi-resistant Staphylococcus spp. (n = 30) and Enterobacteriaceae (n = 26) isolated from dogs referred to a veterinary teaching hospital in Madrid. Results and Discussion: The results indicated the in vitro efficacy of PTSO/PTS against the tested bacterial strains, and 56.7% of Staphylococcus pseudintermedius and 53.8% of Enterobacteriaceae showed sensitivity to PTS and PTSO compared with classic antibiotics. In addition, 50% of S. pseudintermedius strains resistant to erythromycin, ibofloxacin, difloxacin and orbifloxacin and 50% of Enterobacteriaceae strains resistant to tetracycline and doxycycline were sensitive to PTS and PTSO. Although studies are needed to verify their efficacy in vivo, the combined use of PTS and PTSO exhibits promise in enhancing bacterial sensitivity against S. pseudintermedius and Enterobacteriaceae infections, providing a first insight into the potential of both compounds in veterinary practice., Centro para el Desarrollo Tecnológico y la Innovación (España), Depto. de Medicina y Cirugía Animal, Depto. de Genética, Fisiología y Microbiología, Fac. de Veterinaria, TRUE, pub