BackgroundGlobally, antimicrobial resistance (AMR) is a complex public problem, which is mainly fuelled by inappropriate use of antimicrobials. Rational use of antimicrobials is the main strategy for the prevention of AMR, which can be achieved by changing the prescribers’ behavior and knowledge. Hence, this study aimed to assess knowledge and attitude of paramedical students regarding antimicrobial resistance, which helps to rationalize the use of antimicrobials.MethodsAn institutional based cross-sectional study was performed on 323 graduates paramedical students at the University of Gondar, Ethiopia. Participants were invited to complete a self-reported structured questionnaire on hard copy. The data were summarized using summary statistics such as the median. Furthermore, Kruskal Wallis test, at the level of significance of 0.05, was conducted to compare group difference.ResultsAmong 360 eligible paramedical students, 323 (90%) of them participated and most of them were males 202 (62.5%). Nearly 96% of the participants perceived that antimicrobial resistance is a catastrophic and preventable public problem but about half of the participants (55%) had a poor level of knowledge. It was also found that there was a statistically significant knowledge and attitude difference across the department (p-value< 0.0001) and (p = 0.002), respectively. Furthermore, those participants who had a good level of knowledge had greater attitude rank as compared to those who had a moderate and poor level of knowledge (p-value< 0.0001).ConclusionMajority of the participants viewed antimicrobial resistance as a preventable public problem if appropriate strategies are formulated. Nonetheless, most of them had a poor knowledge regarding antimicrobial resistance, and their knowledge and attitude significantly vary across their field of study. This result implicates that improving the students’ level of knowledge about antimicrobial resistance might be an approach to flourish their attitude and to rationalize their antimicrobial use.Electronic supplementary materialThe online version of this article (10.1186/s12879-018-3199-1) contains supplementary material, which is available to authorized users.