IntroductionAntimicrobial resistance (AMR) is a growing global public health concern that was directly responsible for 1.27 million deaths and indirectly associated with 4.95 million deaths in 2019. Antibiotic resistance (ABR) occurs naturally, but its prevalence has been accelerated broadly due to antibiotic misuse and overuse, poor infection control practices, and availability of substandard and falsified medicines. ObjectivesTo assess antibiotic use and knowledge of ABR among graduates of tertiary education in Nigeria and evaluate if tertiary education, academic background (science, social sciences, or the arts) and years post-graduation significantly impact antibiotic use and awareness of antibiotic resistance. ResultDespite having tertiary education, 47.7 % of respondents self-medicate with antibiotics, 54.9 % of respondents don't carry out laboratory investigations before using antibiotics, and 70.1 % reduce or stop taking antibiotics on feeling better. Inconvenience, limited access to doctors and financial constraint were the most common reasons for self-medication. Academic background, age, gender and years post-graduation had no statistically significant effect on self-medication practices (p-value>0.05). Inappropriate antibiotic use for viral-infections like sore throat, cold and flu, and diarrhoea was reported in 50.2, 42.6 and 40.2 % of respondents, respectively. Amoxicillin (54.2 %), metronidazole (48.2 %), and ciprofloxacin (45.6 %) were the most used antibiotics. Pharmacists were the most consulted health professionals before antibiotics were used, and the internet is the most common source of information on antibiotic resistance. Most-respondents, 85.5 %, correctly identified that antibiotic misuse contributes to ABR. ConclusionThe increased knowledge and awareness of ABR among respondents did not translate into proper antibiotic use, emphasizing the need for more stringent regulations on antibiotic dispensing and prescribing. Health sector reforms focused on utilizing technology like telemedicine to improve convenient access to health care, and multiple health insurance schemes to reduce out-of-pocket spending should be prioritized to improve health equity and universal health coverage (UHC).