I am planing an experiment where I need to dose Anaerobic bacteria into mouse. For this I need to make anaerobic suspension from capsulated anaerobic bateria. I do not have anaerobic lab.
How to check the viability of bacteria in suspension?
You will have to study some anaerobic techniques
And, depending on your specific bacteria and their sensitivity to oxygen you might get by with a burning "candle jar", an anaerobic jar and pack, or need a full anaerobic hood.
You will also need to figure out which media to use to enumerate your bacteria
https://universe84a.com/candle-jar/
As per my practical hands-on experience to make an anaerobic suspension from anaerobic bacterial capsules without an anaerobic lab, start by preparing or obtaining pre-reduced anaerobically sterilized (PRAS) media such as BD GasPak™(https://www.bd.com/en-ca/products-and-solutions/solutions/capabilities/bd-gaspak-systems-and-accessories) and adding reducing agents like Oxyrase® (https://www.sigmaaldrich.com/IN/en/product/sigma/sae0013) for Broth. Now, Create an anaerobic environment using an anaerobic jar with a gas pack, such as the BD GasPak™ EZ Anaerobe Container System. Open the capsules within this environment and suspend the contents in the PRAS media, then gently vortex to mix. Store the suspension in a sealed vial within the anaerobic jar until use. To check viability, inoculate a small volume onto anaerobic agar plates like Anaerobe Systems Brucella Blood Agar and incubate anaerobically at 37°C for 24-48 hours, then count the colony-forming units (CFUs). Alternatively, perform a Gram stain, use viability stains such as LIVE/DEAD® BacLight™, or metabolic activity assays like resazurin reduction to assess viability. Maintain strict anaerobic conditions and aseptic techniques throughout the process.
Following these steps mentioned by me, you can get a standard result. Feel free to contact me if you need any further assistance regarding this.
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