• The advantages of the use of discs containing single agents in disc diffusion testing of the susceptibility of Aeromonas salmonicida to potentiated sulphonamides

      Douglas, I.; Ruane, N.M.; Geary, M.; Carroll, C.; Fleming, G.T.A.; McMurray, J.; Smith, P. (Elsevier, 2007)
      The susceptibilities of 106 strains of Aeromonas salmonicida to trimethoprim/sulfamethoxazole (SFT) were determined in two laboratories using the Clinical and Laboratory Standards Institute's M42-A disc diffusion protocols. The data generated by the use of discs containing 25 μg SFT (SFT25) allowed the strains to be placed into two groups. Strains in one group (17 strains) generated no inhibition zones and the zones obtained from the other 89 strains were distributed over a wide range but showed no natural division into separate sub-classes. A further investigation performed by one of the participating laboratories, of the susceptibility of 91 of these 106 strains used discs containing 100 μg sulfmethoxazole (SFM100) and 5 μg trimethoprim (TMP5). Application of normalised resistance interpretation to these data allowed the estimation of epidemiological cut-off values for WT strains of ≥ 9 mm for SFM100 and ≥ 21 mm for TMP5. This investigation demonstrated the presence of three distinct phenotypic classes, one containing strains manifesting wild type susceptibility to both agents, another containing strains manifesting non-wild type susceptibility to both and a third containing strains manifesting wild type susceptibility with respect to TMP but non-wild type with respect to SFM. Analysis demonstrated the inability of SFT25 discs to generate data that allowed the separate identification of strains that were fully susceptible to both TMP and SFM from those that were fully susceptible to TMP but were not fully susceptible to SFM. It is recommended that, in investigation of the susceptibility to potentiated sulphonamides of isolates from diseased fish, separate discs, containing the individual components of the mixture, should be employed.
    • Impact of inter-lab variation on the estimation of epidemiological cut-off values for disc diffusion susceptibility test data for Aeromonas salmonicida

      Smith, P.; Ruane, N.M.; Douglas, I.; Carroll, C.; Kronvall, G.; Fleming, G.T.A. (Elsevier, 2007)
      Two laboratories investigated the susceptibility of 106 Aeromonas salmonicida strains (from Denmark, France, Ireland, Norway and Scotland) to erythromycin, gentamicin, oxytetracycline and oxolinic acid using the disc diffusion protocols (M42-A) published by the Clinical and Laboratory Standards Institute. In studies of susceptibility to florfenicol an additional 15 Canadian strains were included. Comparison of the data generated by the two laboratories demonstrated that for each disc both detected a similar pattern of distribution but that there was a significant numerical difference in the zone sizes they recorded. Analysis of the extent of this lateral shift between the data generated in two laboratories indicated that the application of a single laboratory-independent epidemiological cut-off value for each disc could result in disagreement between the laboratories as to whether a strain should be classified as wild-type or non wild-type. Normalised resistance interpretation was employed to generate epidemiological cut-off values from the data obtained by each laboratory. The use of these laboratory-specific cut-off values resulted in both laboratories achieving complete agreement as to the classification of all strains to all agents.