The absorption and excretion times of oxolinic acid(OX) used in farms as new aquatic antibiotics commonly were evaluated with determination of the effects of water temperature and feeding to parameters by using the bioassay technique. On the same time, antibacterial activity and the complex formation of oxolinic acid with serum proteins of two different fishes were compared to those oxytetracycline(OTC). With more than 10 times lower MIC values than those of OTC in the strains among 13 analyzed fish pathogens. OX did not show the decresed antibacterial activity by the binding of serum proteins in carp and tilapia. It implies more powerful potential of OX as aquatic medicine OTC. The serum concentration of OX after different administrations the oral, i.m., i.v and dipping methods were compared. The higher beginning concentration in serum and faster excretion times were obserbed in i.m. and dipping methods respectively. In the oral and i.m. administration, peak serum concentration after 24－48 hrs and slow excretion times demonstrated in both methods. These pharmacokinetic characteristics similar at $30^{circ}C$ and $20^{circ}C$ water temperature conditions, however, beginning serum concentration of OX in fish dipped in $50mg/ell$ sol after starvation for 2 wks was appeared lower than those of fed fish. It suggests the importance of biological condition of the gill or skin for absorption of antibiotics after dipping administration.