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Although numerous reports have compared the antimicrobial susceptibility of Campylobacter spp., controversy still exists about the use of the E test as an alternative to the agar dilution method suggested by the Clinical and Laboratory Standards Institute. MICs of 8 antimicrobials were determined using the E test and agar dilution methods for 103 Campylobacter jejuni and Campylobacter coli isolates from fresh chicken randomly purchased from stores in 3 southern Ontario counties. Overall, 72.6% of E test MIC values were within 1 log(2) dilution and 95.7% within 2 log(2) dilutions of the corresponding agar dilution MICs. For individual antimicrobials, agreement within 1 log(2) dilution and 2 log(2) dilutions was as follows: ampicillin (n = 103), 90.3% and 98.1%, respectively; chloramphenicol (n = 104), 85.6% and 99%; ciprofloxacin (n = 99), 51.5% and 97.0%; clindamycin (n = 99), 26.3% and 78.8%; erythromycin (n = 99), 52.5% and 96.0%; gentamicin (n = 99), 100% and 100%; nalidixic acid (n = 98), 91.8% and 99.0%; and tetracycline (n = 86), 82.6% and 97.7%. Relative to agar dilution, the E test underestimated the MIC value by a mean of 0.74 (ampicillin), 0.82 (chloramphenicol), 1.44 (ciprofloxacin), 1.94 (clindamycin), 1.40 (erythromycin), 0.21 (gentamicin), 0.94 (nalidixic acid), and 0.20 (tetracycline) log(2) dilutions and by a median of 1 log(2) dilution for all antimicrobials except clindamycin (2), gentamicin (0), and tetracycline (0). Cost analysis, including materials and labor, showed a 39.0% higher cost per analyte for the agar dilution method as compared with the E test. The most relevant advantage of the E test over the agar dilution method is the turnaround time because testing 99 strains by the agar dilution method takes 3.6 times longer compared with the E test using the same number of strains. The E test is an acceptable alternative for antimicrobial susceptibility testing in Campylobacter because it corresponds well with the agar dilution method although being considerably less expensive, is less labor intensive, and is more rapid. However, the relationship between E test and agar dilution MICs must be considered when interpreting E test results.