Isolation and characterization of a novel methanogen Methanosarcina hadiensis sp. nov. from subsurface Boom Clay pore water.

Safe geological disposal of radioactive waste requires a thorough understanding of geochemical conditions in the host formation. Boom Clay is a potential candidate in Belgium, where active methanogenesis has been detected in its deep subsurface, influencing the local geochemistry. However, the pathways driving this process and the characteristics of the methanogenic archaea involved remain unclear. We isolated a distinct archaeal strain from Boom Clay pore water and characterized it geno- and phenotypically. Isolate TD41E1-1 belongs to a novel species of the Methanosarcina genus, for which the name Methanosarcina hadiensis sp. nov. is proposed. TD41E1-1 cells are coccus-shaped, irregularly sized cells enveloped by extracellular polymer substances. Growth and substrate utilization experiments and genomic analysis demonstrated that the strain prefers methylated compounds or hydrogen as substrates for methane production. Although it possesses a complete acetoclastic pathway, no growth was observed in the presence of acetate in the tested conditions. Based on its phylogenetic relation to other known Methanosarcina species and on the presence of c-type cytochromes, it can be concluded that the strain likely occupies an intermediate position between type I and type II Methanosarcina species. These findings provide valuable insights for assessing Boom Clay's suitability for geological disposal of radioactive waste.