Pyelonephritis is a serious condition that is rarely described in horses. In contrast, urinary tract infections are common in humans and small animals, and multi-drug-resistant urinary infections are an emerging threat. In this report, we describe a horse with unilateral pyelonephritis caused by extended-spectrum beta-lactamase-producing bacteria belonging to the Enterobacter cloacae complex. [Correction added on 9 August 2023, after first online publication: The preceding sentence was corrected.] An 11-year-old Swedish warmblood gelding was diagnosed with a cystolith and a cystotomy through an open left para-inguinal approach was performed. Seven days after surgery the horse presented with pyrexia, dullness and colic. Diagnostic testing and renal transabdominal ultrasonography confirmed the presence of a right-sided pyelonephritis. Culture and antimicrobial susceptibility testing revealed a pure growth of extended-spectrum beta-lactamases-producing E. cloacae complex bacteria with resistance against beta-lactams, aminoglycoside and trimethoprim-sulphonamide classes. Treatment included prolonged oral antimicrobials according to susceptibility testing results (enrofloxacin), judicious use of non-steroidal anti-inflammatory drugs, fluid therapy and gastric ulcer prophylaxis. The horse recovered successfully and is currently in good health (follow-up of 5 years). Once the infection resolved, unilateral renal scarring occurred. Multidrug-resistant upper-urinary infections occur in horses and should be considered in a post-surgical patient that develops fever. Early diagnosis, urine bacterial culturing and antimicrobial susceptibility testing were crucial in this case to successful management.