A novel apramycin resistance gene, apmA, was detected on the ca.-40-kb resistance plasmid pAFS11 from bovine methicillin-resistant Staphylococcus aureus (MRSA) of sequence type 398 (ST398). The apmA gene coded for a protein of 274 amino acids that was related only distantly to acetyltransferases involved in chloramphenicol or streptogramin A resistance. NsiI deletion of apmA resulted in a 16- to 32-fold decrease in the apramycin MICs. An apmA-specific PCR identified this gene in one additional bovine and four porcine MRSA ST398 isolates.