Olive flounder is the most important species for the Northeast Asian fish farming industry. However, this species is substantially affected by multiple infectious agents, including Streptococcus parauberis. Evaluation of antibiotics before their application is critical to treat infections and prevent drug resistance. Therefore, in this study, the pharmacodynamics of amoxicillin (AMX) and other antimicrobials against the planktonic-and biofilm-forming bacteria were assessed. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) andTime-kill curve assay were analysed using micro-dilution method. The minimum biofilm eradicating concentration (MBEC) was determined using the Calgary Biofilm device. The effects of temperature, pH, hardness and salinity were detected for both planktonic-and biofilm-forming bacteria. The MIC of AMX ranged from 0.015 to 2 lg/ml, whereas that of cephalexin (CEP), enrofloxacin (ENR) and oxytetracycline (OTC) ranged from 0.125 to 256, 0.125 to >512 and 0.25 to >512 lg/ml respectively. No bacteria were resistant against AMX, while the percentage of resistance to CEP, OTC and ENR were 68.7%, 52.6% and 11.1% respectively. The IC 50 of AMX, CEP, ENR and OTC was 0.03, 0.091, 0.015 and 0.213 lg/ml respectively.The MBEC of amoxicillin against S. parauberis ranged from 0.5 to 16 lg/ml. Higher rates of bacterial growth were obtained at 30°C, pH = 8 and salinity of 7.5-10 ppt.The hardness of the media suppressed the bacterial growth. In conclusion, AMX was found to be effective against both the planktonic and the biofilm forms of the prominent fish pathogen, S. parauberis. K E Y W O R D S biofilm, minimum inhibitory concentration, minimum biofilm eradication concentration, Oliver flounder, Streptococcus parauberis, time-kill curve wileyonlinelibrary.com/journal/are