Agar well technique for antibiotics in animal feeds

The agar well technique compares favorably with the cylinder plate assay in accuracy, sensitivity, and precision. It is more flexible and more rugged, and growth of seed organism is not inhibited. The wells are precision cut with Grafar gel punch assembly with sets (6) of 10, 7, 5, 4, and 3 mm cutters. The wells are easily and rapidly filled with short, disposable Pasteur pipets fitted with rubber bulbs. The smaller wells are filled with capillary pipets. The diameter of the well (independent of volume) appears to be a function of concentration. For every decrease in the diameter size of the well, concentration can be increased, at least to the next higher level of the standard response line. Extracts of chlortetracycline containing as much as 3.2 mu-g/ml can be analyzed if the 3 mm wells are used and with no sacrifice in accuracy or precision. This works especially well for antibiotic premixes. The technique has been used successfully for penicillin, streptomycin, chlortetracycline, oxytetracycline, oleandomycin, and tylosin.