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Staphylococcus aureus,  the leading cause of hospital-acquired infections in the United States, is also pathogenic to the model nematode  Caenorhabditis elegans . The  C. elegans-S. aureus  infection model was previously carried out on solid agar plates where the bacteriovorous  C. elegans  feeds on a lawn of  S. aureus . However, agar-based assays are not amenable to large scale screens for antibacterial compounds. We have developed a high throughput liquid screening assay that uses robotic instrumentation to dispense a precise amount of methicillin resistant  S. aureus  (MRSA) and worms in 384-well assay plates, followed by automated microscopy and image analysis. In validation of the liquid assay, an MRSA cell wall defective mutant, MW2Δ tarO,  which is attenuated for killing in the agar-based assay, was found to be less virulent in the liquid assay. This robust assay with a Z’-factor consistently greater than 0.5 was utilized to screen the Biomol 4 compound library consisting of 640 small molecules with well characterized bioactivities. As proof of principle, 27 of the 30 clinically used antibiotics present in the library conferred increased  C. elegans  survival and were identified as hits in the screen. Surprisingly, the antihelminthic drug closantel was also identified as a hit in the screen. In further studies, we confirmed the anti-staphylococcal activity of closantel against vancomycin-resistant  S. aureus  isolates and other Gram-positive bacteria. The liquid  C. elegans – S. aureus  assay described here allows screening for anti-staphylococcal compounds that are not toxic to the host.

Whole Animal Automated Platform for Drug Discovery against Multi-Drug Resistant Staphylococcus aureus