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We measured methane (CH₄) and stable carbon isotope of methane (δ 13 C-CH₄) concentrations in ambient air and within a red wood-ant (RWA; Formica polyctena ) nest in the Neuwied Basin (Germany) using high-resolution in-situ sampling to detect microbial, thermogenic, and abiotic fault-related micro-seepage of CH₄. Methane degassing from RWA nests was not synchronized with earth tides, nor was it influenced by micro-earthquake degassing or concomitantly measured RWA activity. Two δ 13 C-CH₄ signatures were identified in nest gas: -69‰ and -37‰. The lower peak was attributed to microbial decomposition of organic matter within the RWA nest, in line with previous observations that RWA nests are hot-spots of microbial CH₄. The higher peak has not been reported in previous studies. We attribute this peak to fault-related CH₄ emissions moving via fault networks into the RWA nest, which could originate either from thermogenic or abiotic CH₄ formation. Sources of these micro-seepages could be Devonian schists, iron-bearing "Klerf Schichten", or overlapping micro-seepage of magmatic CH₄ from the Eifel plume. Given the abundance of RWA nests on the landscape, their role as sources of microbial CH₄ and biological indicators for abiotically-derived CH₄ should be included in estimation of methane emissions that are contributing to climatic change.

Can a Red Wood-Ant Nest Be Associated with Fault-Related CH4 Micro-Seepage? A Case Study from Continuous Short-Term In-Situ Sampling