Mycoplasma bovis is considered a major contributor to respiratory diseases in young cattle. Resistant M. bovis isolates have increasingly been reported worldwide due to extensive use of antimicrobials to treat bovine pneumonia. The frequency of isolates resistant to fluoroquinolones varies considerably from one country to another. The MICs of isolates collected in France have only increased from "very low" to "low." The present study was conducted to investigate whether alterations in the quinolone resistance-determining regions (QRDRs) could account for this slight modification in susceptibility. No correlation between QRDR alterations and increased MICs was evidenced in clinical isolates. In addition, all clinical isolates were subtyped, and the tendencies of the different sequence types to develop resistance through mutations in QRDRs under selective pressure in vitro were examined. In vitro, 3 hot spots for mutations in QRDRs (position 83 in GyrA and positions 80 and 84 in ParC) were associated with a high level of resistance when cumulated. We showed that the point mutations in the QRDRs observed in vitro were different (in location and selection rapidity) between the different subtypes. Our in vitro observations were corroborated by the recent detection of a clinical isolate highly resistant to fluoroquinolones (MIC > 16 g/ml) and belonging to the subtype which easily accumulates QRDR alterations in vitro. The current increased prevalence of this subtype in clinical isolates highlights the urgent need to control fluoroquinolone usage in veterinary medicine.
Mycoplasma bovis is a wall-less bacterium responsible for severe infections in cattle, including pneumonia, mastitis, arthritis, and otitis (1). In young cattle, it is now recognized as a major contributor to economic losses associated with bovine respiratory diseases (BRD). The infection pressure of BRD usually peaks 2 to 3 weeks after calves are mingled in fattening units following transportation from their respective birth farms (2). Since no efficient vaccines are available and licensed for use outside the United States (3), efforts to control M. bovis infections often rely on antimicrobial treatments, administered either prophylactically or in the early stages of the disease. Antimicrobials used for the treatment or prevention of BRD usually include broad-spectrum cephalosporins (cefquinome and ceftiofur), extended-spectrum fluoroquinolones (enrofloxacin, danofloxacin, and marbofloxacin), florfenicol, and long-lasting macrolides (tulathromycin, gamithromycin, and tildipirosin) (4). This extensive use of antibiotics has predictably resulted in an increase in resistant isolates over time. In France, it was recently shown that contemporary M. bovis strains had become significantly less susceptible than archival strains to 9 of the 12 antimicrobials tested (5). With regard to fluoroquinolones, the decrease in susceptibility was limited (only 1 dilution of the MIC), and no highly resistant isolates were observed in a set of more than 90 strains. This wa...