Antibiotic use in pets can contribute to antimicrobial resistance (AMR) in both humans and pets. There is a paucity of data on pet owners' knowledge, attitudes, and practices regarding antibiotic use in pets in Asia, despite increasing pet ownership. Therefore, we conducted a study to compare the knowledge, attitudes and practices of antibiotic use and AMR for pets amongst cat and/or dog owners in Singapore. A cross-sectional survey among 1080 pet owners from 16 veterinary clinics between March 2023 and December 2023 evaluated pet owners' knowledge of antibiotic use, understanding of AMR, and antibiotic practices for their pets. Majority of pet owners were female (63.9%), higher educated (86.9%), owned dogs only (62.2%) and currently working (76.4%). Nearly half (46.3%) of all pet owners had poor knowledge of antibiotic use for pets, with cat owners demonstrating poorer knowledge than dog owners (55.2% vs 43.3%, p<
0.001). While only one-third (33.9%) of all pet owners used antibiotics inappropriately, two-thirds (67.1%) demonstrated poor AMR knowledge. Cat owners preferred administering antibiotics in the form of liquids, whilst dog owners in the form of tablets. Pet owners play an important role in antibiotic use in pets. Addressing pet owners' poor knowledge of appropriate antibiotic use and AMR for pets is imperative, to strengthen antimicrobial stewardship efforts via a One Health approach.