Home › Microbiology › Using the Protozoan Paramecium caudatum as a Vehicle for Food-borne Infections in Zebrafish Larvae
Microbiology JoVE (Open Access) Citable · DOI

Using the Protozoan Paramecium caudatum as a Vehicle for Food-borne Infections in Zebrafish Larvae

DOI: 10.3791/58949-v
What you'll learn
  • ✓ Culture Paramecium caudatum and load with foodborne bacteria
  • ✓ Administer bacterial-laden paramecia to zebrafish larvae orally
  • ✓ Quantify bacterial colonization in infected larval zebrafish
  • ✓ Model foodborne pathogenesis using natural predation behavior
Protocol

Zebrafish (Danio rerio) are becoming a widely-used vertebrate animal model for microbial colonization and pathogenesis. This protocol describes the use of the protozoan Paramecium caudatum as a vehicle for food-borne infection in zebrafish larvae. P. caudatum readily internalizes bacteria and get taken up by larval zebrafish through natural preying behavior.

Difficulty
intermediate
Total time
~3–5 days (paramecia culture and bacterial loading) + ~2 hrs (infection and sampling per cohort)
Model organism
Zebrafish (Danio rerio) larvae; Paramecium caudatum
Biosafety
BSL-1

Steps

1
Grow and maintain Paramecium caudatum cultures

Culture P. caudatum under standard conditions and determine appropriate bacterial dose for loading into paramecia without killing the protozoan host.

▶ 00:40
2
Prepare paramecia with internalized bacteria

Load P. caudatum with the target foodborne pathogen, ensuring bacteria are internalized prior to offering paramecia to zebrafish larvae.

▶ 02:23
3
Expose zebrafish larvae to infected paramecia

Allow larval zebrafish to consume bacterial-laden paramecia through natural predation and determine infection success rate or bacterial uptake.

▶ 04:40
4
Assess bacterial colonization in infected larvae

Quantify and visualize bacterial burden within zebrafish tissues following consumption of infected paramecia.

▶ 06:30
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