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Abstract An understanding of the mechanistic drivers of animal‐microbial symbiosis and associations generally requires experiments that manipulate specific symbionts or microbial communities. As part of an ongoing study of the mechanisms that drive microbial communities in coral reef fishes, and specifically the role of cleanerfish in microbial transmission, we tested the effects of the commonly used antibiotics Enrofloxacin and Nitrofurazone on the skin microbiome of three ornamental reef fish species: the four‐eyed butterflyfish, Chaetodon capistratus (Chaetodontidae), the cleanerfish neon goby Elacatinus oceanops (Gobiidae) and the beaugregory damselfish Stegastes leucostictus (Pomacentridae). Our main aim was to characterize dysbiosis prompted by the delivery of the two antibiotics and understand whether both could be used to establish good starting points for microbial transmission experiments. We bathed the fish in antibiotic (or no exposure in the controls), sampled the skin microbiota via swabbing at zero (before treatment) and 3 and 7 days during the treatments, and examined the microbial community using a 16S rRNA gene sequencing approach. Nitrofurazone reduced skin‐associated microbial diversity in all species, whereas the same effect for Enrofloxacin was only seen in S. leucostictus. Although each antibiotic had its own, unique impact in microbial community, all treatments showed positive and negative shifts in the most abundant microbial taxa over time. Moreover, soon after the delivery of both antibiotics, increases in the abundance of opportunistic bacteria or potential pathogens, such as Alteromonas and Vibrio, were observed. Although both antibiotics are effective, Nitrofurazone more successfully reduces microbial diversity and therefore may be more ideal for experiments seeking to disrupt fish microbiomes.