This article describes the installation of an electrochemical system in a natural reservoir to study anode-respiring bacteria. The process involves creating a redox gradient to facilitate bacterial growth and biofilm formation on the anode.
Begin by installing an on-site electrochemical system with the anode in oxygen-free deep water and the cathode in the oxygen-rich surface water of a natural reservoir.
Connect the electrodes with an external circuit to establish a redox gradient that enriches anode-respiring bacteria.
These bacteria, naturally present in anaerobic water, colonize the anode and use outer membrane redox proteins to transfer intracellular electrons to it via extracellular electron transport.
The electrons then flow through the circuit to the cathode, where oxygen serves as the electron acceptor, generating current.
This continuous electron transfer supports bacterial growth and biofilm formation on the anode.
Once the biofilm forms, transfer the anode into a test tube placed underwater to maintain anaerobic conditions and preserve bacterial viability.
Cut the electrode wire while keeping the anode submerged to avoid oxygen exposure.
Seal the tube immediately and store it at a low temperature.
To collect an electrode sample from the natural environment, at least 30 minutes prior to collection, place an uncapped glass test tube and the test tube lid in an anaerobic location.
After placing the bottle into the collection location, make sure that all of the air bubbles have been removed from inside the bottle.
At the end of the cultivation period, use a wire cutter to cut the titanium lead from the electrode and gently collect the electrode sample into the test tube. Cap the test tube within the anaerobic water zone and immediately store the sample at 4 degrees Celsius.
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