This study evaluates the virulence of various strains of pathogenic bacteria using C. elegans as a model organism. By assessing worm survival rates after exposure to different bacterial strains, the relative toxicity of each strain can be determined.
Take a multiwell plate containing suspensions of a non-pathogenic bacterium, serving as a control, and various strains of pathogenic bacteria to evaluate their relative virulence.
The bacteria are suspended in a nutrient-limited medium to prevent overgrowth and to mimic an environment conducive to infection.
Add L4-stage C. elegans worms to each well and incubate the plate under continuous agitation to enhance contact between the bacteria and the worms.
The worms feed on the bacteria, which enter their intestinal tract and colonize it.
The pathogenic bacteria release toxins that damage intestinal tissues, leading to systemic toxicity and eventual death of the worms.
After incubation, count the live worms in each well and generate a graph to assess their survival rates.
Differences in the worm survival rates reflect the relative virulence of each bacterial strain.
After culturing Aeromonas strains and measuring their absorbance as previously described, centrifuge the bacterial broth at 3,500 g’s for 15 minutes. Adjust the bacterial broth with S Medium and add 195 microliters of bacterial broth in the S medium to eight wells of a 96-well plate. Wash the worms off of the ENGM plate with M9 medium. Then, adjust the concentration of the worm solution to five worms per microliter.
Add 5 microliters of the worm solution to each of the wells of the 96-well plate. Incubate the plate on a shaker, and count the number of live worms at 24, 48, and 72 hours.
繁體中文
