Cultivation of Pathogenic Spirochetes to Investigate Skin Infection

Take a skin biopsy from a host infected with a pathogenic spirochete.

In the skin, spirochetes localize within the dermis. The motile spirochetes migrate using the host extracellular matrix to evade the body’s immune responses.

Disinfect the tissue with ethanol and hydrogen peroxide, then transfer it to a petri dish containing saline.

Cut the tissue into pieces, place them in an antibiotic-supplemented prewarmed medium, and incubate.

Antibiotics eliminate the contaminating bacteria, while the spirochetes' outer membrane provides resistance against the antibiotics, allowing their survival.

The spirochete replicates and reaches an exponential growth phase.

Next, dilute the spirochete culture to the desired concentration. Add molten semi-solid agarose, mix, and overlay it onto the solidified nutrient agarose base.

This immobilizes the motile spirochetes, preventing their dispersal while permitting diffusion of nutrients and gases to support growth.

After media solidification, incubate to allow spirochete growth.

The discrete spirochete colonies confirm the spirochete skin infection.

For the cultivation of Borrelia from a skin biopsy, surface-sterilize the skin biopsy with 70% ethanol and hydrog