| Summary: | Background: <i>Acinetobacter</i> spp. have emerged as troublesome pathogens due to their multi-drug resistance. The majority of the work to date has focused on the antibiotic resistance profile of <i>Acinetobacter baumannii</i>. Although <i>A.</i> <i>calcoaceticus</i> strains are isolated in the hospital setting, limited information is available on these closely related species. Methods & Results: The computational analysis of antibiotic resistance genes in 1441 <i>Acinetobacter</i> genomes revealed that <i>A. calcoaceticus</i> harbored a similar repertoire of multi-drug efflux pump and beta-lactam resistance genes as <i>A. baumannii</i>, leading us to speculate that <i>A. calcoaceticus</i> would have a similar antibiotic resistance profile to <i>A. baumannii</i>. To profile the resistance patterns of <i>A. calcoaceticus</i>, strains were examined by Kirby–Bauer disk diffusion and phenotypic microarrays. We found that <i>Acinetobacter</i> strains were moderately to highly resistant to certain antibiotics within fluoroquinolones, aminoglycosides, tetracyclines, and other antibiotic classes. These data indicate that <i>A. calcoaceticus</i> has a similar antibiotic resistance profile as <i>A. baumannii</i> ATCC 19606. We also identified that all <i>Acinetobacter</i> species were sensitive to 5-fluoroorotic acid, novobiocin, and benzethonium chloride. Conclusion: Collectively, these data provide new insights into the antibiotic resistance in <i>A. calcoaceticus</i> and identify several antibiotics that could be beneficial in treating <i>Acinetobacter</i> infections.
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