Gentamicin resistance in dairy and clinical enterococcal isolates and in reference strains

Enterococci isolated from Portuguese dairy products (milk and cheese) and clinical settings (hospitals and veterinary clinics), together with reference strains from the genus Enterococcus, were screened for low- and high-level gentamicin resistance using the standard disc diffusion method (10 and 120 microg gentamicin discs). MICs were also determined using both the macrodilution method and the Etest. Four genes [aac(6')-Ie-aph(2")-Ia, aph(2")-Ib, aph(2")-Ic and aph(2")-Id] responsible for high- and mid-level gentamicin resistance were sought using PCR. Although enterococci generally are regarded as being intrinsically resistant to low levels of gentamicin, results revealed that many dairy enterococci (around 30% of the isolates used) are not intrinsically resistant to gentamicin, showing MICs of < or = 4 mg/l. High-level gentamicin resistance was not detected in any of the dairy isolates studied, except for aph(2")-Ib, which was found in one. Therefore, gentamicin resistance should be monitored in dairy enterococci, although it does not seem to be a problem at present. In contrast, all clinical isolates studied were, as expected, intrinsically resistant to low levels of gentamicin, presenting MICs > 8 mg/l. Fifteen percent of these clinical isolates showed high-level gentamicin resistance (MICs > 512 mg/l), with the bifunctional gene aac(6')-aph(2") being detected in four of them. However, discs with gentamicin 120 microg failed to detect some isolates with high-level gentamicin resistance.