South American camelids (SACs), particularly llamas (Lama glama) and alpacas (Vicugna pacos) are gaining popularity in Europe. Initially valued for their fiber and land management capabilities, these animals are now also kept for animal therapy, outdoor activities, and as companion animals. Despite their close interactions with humans and other animals, there is limited research on the transmission of microbes or antimicrobial resistance genes from SACs. This study aimed to survey the fecal carriage of extended-spectrum beta-lactamase ESBL-producing Escherichia coli (ESBL- E. coli) in SACs. A questionnaire was administered on-site to the farmers to survey management and biosecurity measures. Twelve farms from northern Italy (Lombardy, Piedmont, Veneto, and Emilia-Romagna) participated in the study. Fecal samples were analyzed to identify ESBL- E. coli and subjected to bacteriological culture on CHROMagar™ ESBL plates. Isolate identification was accomplished by MALDI-TOF MS, then subjected to the double-disk synergy test (DDST) and examined for 3 ESBL-encoding genes (bla