Amoeba-Based Enrichment of Giant Viruses from an Environmental Water Sample

Take a multi-well plate containing amoebae cultured in starvation medium.

The starvation medium contains antibiotics and antifungal agents that limit microbial growth while keeping amoebae viable without multiplying or forming dormant cysts.

Take a tube containing the environmental water sample and vortex it to release viruses from suspended solids and debris.

Add the water sample containing the viruses to the amoebae.

Place the plate in a bag with a wet compress to prevent evaporation, and incubate.

During incubation, the amoebae engulf the giant viruses by phagocytosis.

Once internalized, the virus releases its genome into the amoeba’s cytoplasm.

A cytoplasmic viral factory forms around the viral genome, where it uses host machinery to replicate and produce viral proteins that assemble into new virions.

This causes the amoeba cells to round up and lyse, releasing new virions into the medium, enriching the giant viruses for downstream detection.

After collecting dirty water samples according to the text protocol, prepare the host cell, V. vermiformis by adding the amoebae to a 75 square centimeter cell culture flask with 30 milliliters of protease peptone yeast extract glucose, or PYG medi