Selective Cleaning of Caenorhabditis Worms for Enrichment of Intestinal Microbiota

Take a culture plate with Caenorhabditis larvae harboring the bacteria of interest colonizing their intestinal lumen.

The plate contains a known bacterial strain as food. Starve the worms so they consume all the bacteria and enter a dormant stage.

Add a medium and transfer the worms to a tube.

Centrifuge and discard the supernatant.

Add a detergent solution, agitate to remove surface-adhered bacteria, centrifuge again, and discard the supernatant.

Repeat the wash, then transfer the worms to a fresh culture plate.

Allow crawling to remove loosely bound food bacteria from the surface and intestinal lumen while retaining the bacteria of interest that stably colonize the intestine.

Repeat the transfer and crawling step to ensure complete removal of food bacteria.

Add the medium, transfer the worms to a watch glass, and dissect to expose their intestines.

Dissect the intestines containing the bacteria of interest, collect them in water-filled tubes, and freeze for future use.

For starving the worms and reducing the number of OP50-1 bacteria, incubate the nematodes at 20 degrees Celsius for three to four days. Add 5 milliliters of the M9 media to the plate of recently starved worms, and then trans