Muscle Necrosis Assay in C. elegans Induced by a Pathogenic Aeromonas Strain

Begin with a nutrient-rich nematode growth medium plate containing a lawn of the pathogenic bacterium Aeromonas.

Transfer L4-stage C. elegans worms expressing muscle-specific GFP onto the bacterial lawn and incubate.

The worms ingest bacteria along with nutrients. This allows the bacteria to reach and colonize the intestine.

Using a specialized secretion system, Aeromonas delivers virulence factors into the worm’s intestinal cells.

These virulence factors diffuse to the body wall, where they disrupt muscle, causing structural damage and necrosis.

Transfer the worms daily to a fresh plate with a bacterial lawn to maintain continuous pathogen exposure.

Place the worms onto an agarose gel supplemented with an anesthetizing agent to anesthetize them, then place a coverslip.

Using a GFP filter, capture fluorescence images at regular intervals.

The progressive loss of the GFP signal indicates muscle disruption in C. elegans, representing Aeromonas-induced muscle necrosis during host-pathogen interaction.

After culturing Aeromonas strains and adjusting the absorbance according to the text protocol, spot and spread the bacterial broth on NGM plates.

After this, transfer 50 L4 worms to each NGM plate