Inhibition tests for detection and presumptive identification of tetracyclines, beta-lactam antibiotics and quinolones in poultry meat

Abstract

A combination of three plates, seeded with strains of Micrococcus luteus, Bacillus cereus or Escherichia coli, can be used for detection of residues of betalactam antibiotics, tetracyclines and fluoroquinolones. The sensitivity of each plate is optimal for only one of these groups, resulting in detection limits (LOD) lower than the corresponding maximum residue limits (MRL) and in distinct inhibition patterns typical for each antibiotic family. Beta-lactam antibiotics such as penicillin G, ampicillin and amoxicillin give only inhibition zones on the plate with M. luteus. Tetracyclines are detected up to the MRL level with B. cereus, and fluoroquinolones with E. coli. The LODs of the antibiotics tested were as follows: penicillin G (PENG) 0.9 ng, ampicillin (AMPI) 0.6 ng and amoxicillin (AMOX) 1.0 ng on the plate with M. luteus; tetracycline (TET) 4 ng, oxytetracycline (OXY) 3 ng, doxycycline (DOX) 0.6 ng, and chlortetracycline (CHL) 0.3 ng on the plate with B. cereus; enrofloxacin (ENRX) 1.5 ng, ciprofloxacin (CIPX) 0.5 ng and flumequine (FLUM) 1.5 ng on the plate with E. coli. The combination of plates enables the laboratory to select appropriate chromatographic techniques for identification and quantification of the residues. On the other hand, the three groups can also be detected on one plate seeded with Bacillus subtilis, although the limits of detection are higher: PENG 0.4 ng, AMPI and AMOX 3 ng; TET 5 ng, OXY 8 ng, DOX 1 ng, CHL 0.5 ng, ENRX 4 ng, CIPX 10 ng and FLUM 4 ng. The test was applied to 228 broiler fillets and to 27 turkey thighs, originating from different poultry slaughterhouses. Nineteen broiler fillets contained inhibiting substances. The positive results of the inhibition tests were confirmed with a chromatographic technique. Doxycycline residues were found in 16 samples and amoxicillin in two.