Simple SummaryRabbits are the second most common specialty pet among households in Europe and the USA. However, research on antimicrobial resistance (AMR) in pet rabbits is very scarce. Therefore, scientific data on AMR in pet rabbits is urgently needed as a guide for veterinarian clinicians to optimize antibiotic use in rabbits for reducing the selection of antibiotic resistance. In addition, antimicrobial stewardship programs should be conducted to educate rabbit owners not to misuse antibiotics on their pets as it may put their own health at risk. This paper aims to provide an overview of the current state of AMR in rabbits attended to in veterinary clinics distributed in Spain to highlight the importance of addressing AMR under the One Health approach.AbstractResearch on antimicrobial resistance (AMR) in pet rabbits is very scarce. The aim of this study was to provide an overview of the current state of AMR in rabbits attended to in veterinary clinics distributed in Spain. Records of 3596 microbiological results of clinical cases submitted from 2010 to 2021 were analyzed.Staphylococcusspp. (15.8%),Pseudomonasspp. (12.7%),Pasteurellaspp. (10%),Bordetellaspp. (9.6%) andStreptococcusspp. (6.8%) were the most frequently diagnosed agents. Enterobacteriaceae, principallyEscherichia coli,Klebsiella pneumoniaeandEnterobacter cloacae, accounted for about 18% of the cases and showed the highest proportion of multi-drug resistance (MDR) isolates, with 48%, 57.5% and 36% of MDR, respectively. Regarding the antimicrobial susceptibility testing for a number of antimicrobial categories/families, the largest proportion of isolates showing resistance to a median of five antimicrobial categories was observed inP. aeruginosa,Stenotrophomonas maltophiliaandBurkolderiaspp. In contrast, infections caused byStaphylococcus,Streptococcusspp. andPasteurella multocidawere highly sensitive to conventional antimicrobials authorized for veterinary use (categories D and C). The emergence of AMR major nosocomial opportunistic pathogens such asP. aeruginosa, S. maltophiliaandK. pneumoniaein pet rabbits can represent a serious public health challenge. Consequently, collaboration between veterinarians and human health professionals is crucial in the fight against antimicrobial resistance, to optimize, rationalize and prudently use antimicrobial therapies in domestic animals and humans.