There is limited published information regarding antimicrobial use (AMU) and antimicrobial resistance (AMR) in aquaculture. Our objective was to determine the opinions of aquaculture-allied professionals around the world on the frequency of AMU and AMR in common aquatic species. The study questionnaire included five sections: respondent demographics, extent of AMU in aquaculture, frequency of observations of AMR in aquaculture, AMR monitoring and surveillance and antimicrobial susceptibility testing in various jurisdictions. It was administered in English and Spanish to 604 professionals in 25 countries and with varying expertise in aquaculture. The response rate was 33% (199/604). Over half of the participants had >10 years of experience in aquaculture: 70% (140/199) were involved in fish health/clinical work and their primary experience was with salmon, tilapia, trout, shrimp (including prawn) and/or catfish. Tetracycline use was reported by 28%, 46%, 18%, 37% and 9% of respondents working with catfish, salmon, tilapia, trout and shrimp, respectively. Resistance to tetracycline in one or more species of bacteria was reported as ‘frequent-to-almost always’ for the same aquaculture species by 39%, 28%, 17%, 52% and 36% of respondents, respectively. ‘Frequent-to-almost always’ use of quinolone was reported by 70% (32/46) and 67% (8/12) of respondents from the United States and Canada, respectively, where quinolone products are not approved for aquaculture, and extra-label fluoroquinolone use is either prohibited (United States) or discouraged (Canada). Similar frequencies of quinolone use were also reported by the majority of respondents from Europe [70% (7/10)] and Asia [90% (9/10)] where labelled indications exist. This baseline information can be used to prioritize research or surveillance for AMU and AMR in aquaculture.