To investigate the effects of herring oil and unsaturated fatty acids on the virulence of S. aureus MSSA 6538, we used a nematode survival assay as previously described (Kim et al., 2016 (link)) with slight modification. In brief, S. aureus cells were incubated with or without herring oil, DHA, or EPA (2, 5, or 20 μg/ml) at 37°C for 24 h and synchronized adult C. elegans fer-15;fem-1 nematodes were added into single wells of 96-well plate containing cultivated S. aureus cells. Approximately, 30 nematodes were allowed to feed on the cultured S. aureus MSSA 6538 at 25°C for 1 day.
For the cytotoxicity assay, 110 ± 10 nematodes were added into single well of 96-well plates containing M9 buffer and solutions of the compounds were added to final concentrations of 20 or 100 μg/ml at 25°C for 1 day. Then, nematodes were scored as alive or dead using an iRiSTM Digital Cell Imaging System (Logos Bio Systems, South Korea). At least three independent experiments were conducted using quadruplicate wells.
For the cytotoxicity assay, 110 ± 10 nematodes were added into single well of 96-well plates containing M9 buffer and solutions of the compounds were added to final concentrations of 20 or 100 μg/ml at 25°C for 1 day. Then, nematodes were scored as alive or dead using an iRiSTM Digital Cell Imaging System (Logos Bio Systems, South Korea). At least three independent experiments were conducted using quadruplicate wells.
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