The direct connection between consumption of Arcobacter contaminated water and human diseases has not been established yet, although it is likely that water may play an important role in the Arcobacter transmission to humans. Fluoroquinolones are potential drugs to treat infections due to these organisms. However, there is evidence of increasing resistance to these antimicrobial agents. This resistance may occur due mutations in a quinolone resistance-determining region (QRDR) of the gyrA gene. In this study, Arcobacter susceptibility to fluoroquinolones was determined by disc diffusion tests and E-test strips. In addition, to elucidate the mechanism of quinolone resistance, we carried out sequence determination and analysis of the QRDR of their gyrA gene. Among the 60 isolates tested, 14 were considered to be resistant to quinolones by disc diffusion tests and E-test strips. The sequencing of the PCR products revealed a mutation in position 254 of the gyrA gene in the 14 resistant A. butzleri isolates. This C-254 to T mutation could be the cause of quinolone resistance as this change was absent in all the susceptible isolates.