Modeling Intestinal Infection in Zebrafish Larvae Using Foodborne Bacteria

Begin with a multiwell plate containing paramecia pre-infected with pathogenic E. coli expressing fluorescent proteins.

Add anesthetized zebrafish larvae and co-incubate them under daylight conditions to mimic the natural feeding environment.

During incubation, the larvae regain consciousness and prey on infected paramecia.

Once ingested, the paramecia are digested, releasing E. coli into the intestinal tract.

Bacterial colonization progresses from the anterior intestine to the posterior intestine, mimicking a foodborne infection.

Next, wash the larvae in a medium supplemented with an anesthetic agent to re-anesthetize them.

Then, place the larvae in low-melting-point agarose within a fluorescent imaging plate.

Using a stereomicroscope, align the zebrafish and incubate to allow the agarose to solidify.

Overlay with a nutrient-rich medium supplemented with an anesthetic agent.

Using a fluorescence microscope, observe the larvae for the distribution of fluorescent E. coli to evaluate the progression of bacterial infection

For food-borne infection of the zebrafish, dilute the paramecia samples to a two times ten to the five paramecia per milliliter of E3 medium concentration and add 3 millilite