Mycobacterium abscessus (Mabs) is intrinsically resistant to many antimycobacterial antibiotics, which presents serious problems in therapy. Here we describe the development of a novel phenotypic-based microscopic and computerized imaging drug screening approach. A pilot screen of 568 compounds from two libraries identified 17 hits. Eleven of these compounds are described for the first time as active against Mabs. The impact of growth media on these compounds' activity was tested revealing that cation-adjusted Mueller Hinton (MHII) broth supports better growth of actively replicating Mabs and improves the associated compounds activity.
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