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2.3.2.1. Micro-dilution method. This method has been adopted from NCCLS M26-T (Food and Drug Administration, 1991); (National Committee for Clinical Laboratory Standards, 1999), with modifica- tions (Hacek et al., 1999). In short: 100 µl antibiotic solutions were pipetted into micro-titer plates by twofold dilutions. 100 µl freshly grown bacteria were added to the wells in a density of 2.5¡¤105 CFU (final concentration per well). Micro-titer plates were incubated overnight (≙ MBC/L) or 4 h (≙ MBC/S). CFU was determined by diluting each well in tenfold dilutions. From each dilution, aliquots were transferred on agarplates and incubated overnight. On the following day, the number of colonies was evaluated and the initial CFU/well retrospectively calculated. |
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