MRSA is an important antimicrobial-resistant pathogen reported in a wide range of animal species; however, information regarding its occurrence in captive large felids remains limited. This report describes the detection and characterization of MRSA associated with a dental abscess in a captive jaguar (Panthera onca) from a zoological collection in Romania. A 10-year-old male jaguar presented with clinical signs suggestive of a localized oral infection. Samples collected from the dental lesion, nasal mucosa, external ear canal, and skin surface were subjected to bacteriological examination. Bacterial isolates were identified using conventional microbiological methods and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). Antimicrobial susceptibility testing was performed using the VITEK^® 2 system, while molecular confirmation was achieved by PCR targeting the nuc, mecA, and mecC genes. All isolates were identified as S. aureus and demonstrated a methicillin-resistant phenotype. PCR confirmed the presence of the species-specific nuc gene and the methicillin-resistance determinant mecA, while mecC was not detected. The isolates exhibited resistance to multiple antimicrobial classes, including β-lactams, fluoroquinolones, and macrolides, while remaining susceptible to linezolid, vancomycin, teicoplanin, rifampicin, and tigecycline. This case documents the occurrence of MRSA in a captive jaguar and highlights the value of integrated microbiological and molecular investigations for the diagnosis and characterization of resistant bacterial infections in zoological species.