Growth of anaerobic methane oxidizing archaea and sulfate reducing bacteria in a high pressure membrane-capsule bioreactor

Anaerobic methane oxidizing communities of archaea (ANME) and sulfate reducing bacteria (SRB) grow slowly, which limits physiological studies. High methane partial pressure was previously successfully applied to stimulate growth, but it is not clear how different ANME subtypes and associated sulfate reducing bacteria (SRB) are affected by it. Here, we report growth of ANME/SRB in a membrane-capsule bioreactor inoculated with Eckernförde Bay sediment that combines high pressure incubation (10.1 MPa methane) and thorough mixing (100 rpm) with complete cell retention by a 0.2 µm membrane. Results were compared to previously obtained data from an ambient-pressure (0.101 MPa methane) bioreactor inoculated with the same sediment. Labelled-methane oxidation rates were not higher at 10.1 MPa, likely because measurements were done at ambient pressure. The subtype ANME-2a/b was abundant in both reactors, but subtype ANME-2c was only enriched at 10.1 MPa. SRB at 10.1 MPa mainly belonged to the SEEP-SRB2, Eel-1 group and Desulforomonadales and not to the typically found SEEP-SRB1. Increase of ANME-2a/b occurred in parallel with increase of SEEP-SRB2 which was previously only found associated with ANME-2c. Our results imply that the syntrophic association is flexible and that methane pressure and sulfide concentration influence growth of different ANME-SRB consortia. We also studied the effect of elevated methane pressure on methane production and oxidation by a mixture of methanogenic and sulfate-reducing sludge. Here, methane oxidation rates decreased and were not coupled to sulfide production, indicating trace methane oxidation during net methanogenesis and not anaerobic methane oxidation, even at high methane partial pressure.