TABLE 3.
Strain | MIC (μg/ml)a
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AMP | OXA | MCIPC | NAF | CAR | SBPC | CXM | FAM | CTX | CMZ | ATM | FRPM | DOCb | SDSb | NOVb | ERMb | TPPb | |
TG1 | 1.56 | >100 | 200 | >400 | 1.56 | 1.56 | 1.56 | 0.39 | 0.025 | 0.78 | 0.05 | 0.39 | >40,000 | >400 | 50 | 50 | 200 |
KAM3, KAM3/pHSG398 or pHSG399 | 0.78 | 0.39 | 0.78 | 1.56 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 625 | 50 | 0.78 | 1.56 | 3.13 |
KAM3/pHSGacrAB | 1.56 | >100 | 100 | 200 | 1.56 | 1.56 | 1.56 | 0.39 | 0.025 | 0.78 | 0.05 | 0.39 | >40,000 | >400 | 25 | 12.5 | 200 |
KAM3/pHSGacrD | 0.78 | 12.5 | 12.5 | 50 | 25 | 50 | 0.39 | 0.20 | 0.012 | 0.78 | 0.39 | 0.10 | >40,000 | >400 | 6.25 | 1.56 | 3.13 |
KAM3/pHSGacrEF | 0.78 | 50 | 100 | 200 | 1.56 | 1.56 | 0.78 | 0.39 | 0.025 | 0.78 | 0.05 | 0.10 | >40,000 | >400 | 25 | 12.5 | 200 |
KAM3/pHSGmdtABC | 0.78 | 3.13 | 6.25 | 6.25 | 6.25 | 12.5 | 0.20 | 0.10 | 0.012 | 0.39 | 0.10 | 0.10 | 20,000 | 200 | 6.25 | 1.56 | 1.56 |
KAM3/pHSGmdtEF | 0.78 | 12.5 | 12.5 | 25 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 20,000 | 200 | 0.78 | 12.5 | 25 |
KAM3/pHSGemrABc | 0.78 | 0.39 | 0.78 | 1.56 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 2,500 | 100 | 3.13 | 1.56 | 1.56 |
KAM3/pHSGemrKY | 0.78 | 0.39 | 0.78 | 1.56 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 1,250 | 50 | NDe | 1.56 | ND |
KAM3/pHSGmacAB | 0.78 | 0.39 | 0.78 | 1.56 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 625 | 50 | 0.78 | 12.5 | 1.56 |
KAM3/pHSGmdfA | 0.78 | 0.39 | 0.78 | 1.56 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 625 | 50 | 0.78 | 1.56 | 12.5 |
KAM3/pHSGemrEd | 0.78 | 0.39 | 0.78 | 1.56 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 625 | 50 | 0.78 | 1.56 | 3.13 |
KAM3/pHSGmdtK | 0.78 | 0.39 | 0.78 | 1.56 | 1.56 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 625 | 50 | 0.39 | 1.56 | 25 |
TG1ΔtolC | 0.78 | 0.39 | 0.78 | 0.78 | 0.78 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 313 | ND | ND | ND | ND |
TG1ΔtolC/pHSGacrAB | 0.78 | 0.39 | 0.78 | 0.78 | 0.78 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 313 | ND | ND | ND | ND |
TG1ΔtolC/pHSGacrD | 0.78 | 0.39 | 0.78 | 0.78 | 0.78 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 313 | ND | ND | ND | ND |
TG1ΔtolC/pHSGacrEF | 0.78 | 0.39 | 0.78 | 0.78 | 0.78 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 313 | ND | ND | ND | ND |
TG1ΔtolC/pHSGmdtEF | 0.78 | 0.39 | 0.78 | 0.78 | 0.78 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 313 | ND | ND | ND | ND |
TG1ΔtolC/pHSGmdtABC | 0.78 | 0.39 | 0.78 | 0.78 | 0.78 | 1.56 | 0.20 | 0.10 | 0.012 | 0.39 | 0.05 | 0.10 | 313 | ND | ND | ND | ND |
MIC determination was repeated at least three times. Values in bold differ from the KAM3/pHSG398 or 399 control strain value by ≥4-fold. MIC values of benzylpenicillin (12.5 μg/ml for all strains listed in the table), cephalothin (1.56 μg/ml), cefsulodin (12.5 μg/ml), ceftazidime (0.10 μg/ml), flomoxef (0.025 μg/ml), and imipenem (0.05 μg/ml) were not changed by overexpression of any drug exporters tested in this study. Abbreviations: AMP, ampicillin; SBPC, sulbenicillin; CTX, cefotaxime; CMZ, cefmetazole; DOC, deoxycholate; SDS, sodium dodecyl sulfate; NOV, novobiocin; ERM, erythromycin; TPP, tetraphenylphosphonium bromide.
These compounds were used to confirm the overproduction of the pumps. Additionally, CCCP (carbonyl cyanide m-chlorophenylhydrazone) and acriflavine were used to confirm the overproduction of EmrAB and EmrE, as described below.
pHSGemrAB conferred CCCP resistance (MIC, 6.25 μg/ml for KAM3 versus 50 μg/ml for KAM3/pHSGemrAB).
pHSGemrE conferred acriflavine resistance (MIC, 12.5 μg/ml for KAM3 versus 200 μg/ml for KAM3/pHSGemrE).
ND, not determined.