The fluoroquinolone antibacterial family is a relatively recent group of bactericidal compounds, generally characterized by efficacy against a wide spectrum of bacterial organisms and exhibiting minimal adverse effects in treated patients. The fluoroquinolones are widely prescribed in both human and veterinary medicine, though in veterinary medicine in the USA there are currently only two approved compounds, enrofloxacin (Baytril, Bayer Animal Health, Shawnee Mission, KS) and sarafloxacin (SaraFlox, Abbott Laboratories, North Chicago, IL), both with limited species and disease label approvals. Currently, there are no approved fluoroquinolone antibacterials to treat bacterial infectious diseases in cultured fish species. Enrofloxacin was administered to juvenile Atlantic salmon as a single bolus via intraarterial (i.a.), intraperitoneal (i.p.), intramuscular (i.m.), or oral gavage routes of administration. The drug was administered via the first three routes to achieve a dose of 10 mg/kg, and via oral gavage to achieve both 10 (p.o.-10) and 5 (p.o.-5) mg/kg doses. Two-compartment model kinetics were observed with elimination of half-lives (t1/2) of 130.6, 34.32, 84.98, 105.11, and 48.24 h, area under the drug concentration-time curves (AUC) of 84.3, 75.31, 55.61, 41.68, and 38.81 micrograms x h/mL, and bioavailabilities (F) of 100, 89.34, 65.97, 49.44, and 46.04% (i.a., i.p., i.m., p.o.-10, p.o.-5, respectively). All administration routes at 10 mg/kg were found to yield comparable drug concentration-time curves for multiple tissue, indicating no distinct advantage of using one route over another from a kinetics perspective. Finally, the 5 mg/kg dose (p.o.-5) yielded comparable multiple tissue drug concentration-time curves to the 10 mg/kg dose (p.o.-10), providing pharmacokinetic evidence to justify therapeutic efficacy trials with the lower dose.