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. In this study, we report lipid biomarker patterns and phylogenetic identitiesof key microbial communities mediating anaerobic oxidation of methane (AOM)in active mud volcanoes (MVs) on the continental slope of the CanadianBeaufort Sea. The carbon isotopic compositions ( δ13C ) ofsn-2- and sn-3-hydroxyarchaeol showed the highly 13C -depleted values ( − 114 ‰ to − 82 ‰)associated with a steep depletion in sulfate concentrations within 0.7 m ofsediment depths. This suggested the presence of methanotrophic archaeainvolved in sulfate-dependent AOM, albeit in a small amount. The ratio ofsn-2-hydroxyarchaeol to archaeol (> 1) and operationaltaxonomic units (OTUs) indicated that the anaerobic methanotrophic archaea (ANME) clades ANME-2c and ANME-3were involved in AOM. Higher δ13C values of archaeoland biphytanes (BPs; - 55.2 ± 10.0  ‰ and - 39.3 ± 13.0  ‰, respectively) suggested that archaeal communities were alsoassimilating AOM-derived inorganic carbon. Furthermore, the distinctdistribution patterns of methanotrophs in the three MVs appears to beassociated with varying intensities of ascending gas fluids. Consequently,our results suggest that the niche diversification of active mud volcanoeshas shaped distinct archaeal communities that play important roles in AOM inthe Beaufort Sea.

Biogeochemical evidence of anaerobic methane oxidation on active submarine mud volcanoes on the continental slope of the Canadian Beaufort Sea