Feb 08, 2026
Preparation and Handling of Anaerobic Bacteria for Mouse Oral Gavage
- Anastasiya Moiseyenko1
- 1California Institute of Technology
- Faecalibacterium prausnitzii improves motor deficits in a-synuclein overexpressing mice
Protocol Citation: Anastasiya Moiseyenko 2026. Preparation and Handling of Anaerobic Bacteria for Mouse Oral Gavage. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgboqw1lpk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 08, 2026
Last Modified: February 08, 2026
Protocol Integer ID: 242881
Keywords: oral administration, gavage, mouse, anaerobic, bacteria, culture, anaerobic chamber, gaspack, anaeropack, handling of anaerobic bacteria, bacterial culture from an anaerobic chamber, oral administration of bacteria, anaerobic bacteria, transfer of anaerobic, mouse oral gavage, bacterial culture, mouse oral gavage this protocol, preparation
Funders Acknowledgements:
Aligning Science Across Parkinson’s
Grant ID: ASAP-020495
Aligning Science Across Parkinson’s
Grant ID: ASAP-000375
Heritage Medical Research Institute
Grant ID: n/a
Abstract
This protocol outlines the set up and transfer of anaerobic bacterial culture from an anaerobic chamber for the oral administration of bacteria to animals.
Materials
anaerobic chamber
cryotubes
parafilm
grow flasks
1ml slip-tip syringes
plastic gavage tubes
GasPack box (Anaerobe systems)
AnaeroPack - Anaero sachets (MCG)
pre-reduced 50% glycerol
pre-reduced PBS with 1.5% NaHCO3
Troubleshooting
Bacterial growth and aliquots
Grow anaerobic bacteria inside an anaerobic chamber (80% nitrogen, 10% carbon dioxide, and 10% hydrogen) at 37℃ to an approximate optical density (600nm) of 1
For extremely oxygen sensitive bacteria, wrap growth flasks with parafilm prior to removing from anaerobic chamber
Centrifuge culture and resuspend in sterile pre-reduced 50% glycerol to an approximate Colony-Forming Unit (CFU) density of 108/mL inside the chamber
Aliquot into cryotubes
For extremely oxygen sensitive bacteria, wrap cryotubes with parafilm prior to removing from anaerobic chamber
Store aliquots at -20℃ for short-term storage, -80℃ for long-term storage
Preparation for gavage
Thaw desired aliquots while preparing gavage supplies
Attach gavage needles to syringes (inside a hood if sterility is of concern) and place inside GasPack box
Place an AnaeroPack - Anaero sachet inside GasPack box, DO NOT OPEN the sachet until inside the anaerobic chamber
Transfer supplies and aliquots into anaerobic chamber
Transfer aliquots to conical tubes and centrifuge to remove glycerol
Resuspend culture in pre-reduced PBS with 1.5% NaHCO3
Draw up culture into gavage syringes
Place ready gavage syringes and opened AnaeroPack-Anaero sachet in GasPack box, seal tightly
Remove from chamber and transport to animal facility
For extremely oxygen sensitive bacteria, open the GasPack box as minimally as possible during gavage process and administer gavage as quickly as safely possible to animals to reduce oxygen exposure times