Working with Snodgrassella alvi

Snodgrassella alvi is a member of the honey bee (Apis mellifera) gut core microbiota that can be cultivated in vitro. Its isolation and characterization were recently carried out by Waldan Kwong in the laboratory of Dr. Nancy Moran and are described in depth in here.

In vitro culture conditions

Plate Growth

Snodgrassella alvi strain wkB2 (type strain) is generally cultured at 35°C and 5% CO₂. It grows robustly on agar plates after 48 hours on the following media:

• Heart Infusion Agar
• Heart Infusion Agar supplement with Sheep's Blood
• Columbia Agar supplement with Sheep's Blood (CBA)
• R2 Agar

Liquid Media

Snodgrassella alvi can be cultivated in the following liquid media (same general culture conditions as above, without shaking):

• Insectagro (proprietary media)
• Salt-free LB
• R2 Liquid Media

Conjugation

Snodgrassella alvi can be conjugated using E. coli donor strains and mobilizable rsf1010-based plasmids. Refer to the Bee Microbiome Toolkit paper for full description of working with this species.
Perform conjugation as described here: Conjugation, with the following specifics: use 10:1 ratio of S. alvi to donor E. coli pooled on CBA plates. Incubate overnight, then resuspend and plate on selective CBA plates.