Preprints
https://doi.org/10.5194/egusphere-2025-236
https://doi.org/10.5194/egusphere-2025-236
03 Feb 2025
 | 03 Feb 2025
Status: this preprint is open for discussion and under review for Biogeosciences (BG).

Anaerobic biodegradation of Miocene lignites from an opencast mine by autochthonous microorganisms stimulated under laboratory conditions

Michal Bucha, Piotr Siupka, Anna Detman-Ignatowska, Sushmita Deb, Agnieszka Bylina, Daria Kaim, Aleksandra Chojnacka, Dominika Kufka, Dominika Lewicka-Szczebak, Anna Sikora, and Leszek Marynowski

Abstract. The supplementation and provision of appropriate nutrients to microorganisms, which are often lacking in the natural environment are essential and critical for microbial growth. One such element is nitrogen, most of which is found in the Earth's atmosphere. In this study, we present evidence of nitrogen processing and anaerobic N2-fixation by microorganisms naturally present in sedimentary organic matter. Miocene detritic lignite from the opencast mine was incubated under anaerobic conditions in the dark (headspace atmosphere 85 % N2, 10 % CO2, 5 % H2) for three years. The natural microbial community of these coal materials was stimulated for growth through the addition of trace elements, vitamins, and carbon-bearing compounds such as yeast extract, nutrient broth, methanol, and sodium acetate. A visual indicator of microbial activity was observed as the color of the fermentation solutions changed over time: from colorless to light yellow (after 3 months), dark brown (after 6 months), and finally black (after more than 1 year). This progression suggests the dissolution of fulvic and humic acids. At the end of the cultivation period, the total nitrogen (TN) and total inorganic nitrogen (TIN) contents in the solutions were significantly reduced whereas in incubations with sodium acetate, total organic nitrogen (TON) content significantly increased compared to the initial levels. In most cases, total carbon (TC) content significantly increased due to biodegradation, except for the incubations where methanol was added. A GC-MS analysis of the total extracts from lignite revealed that the main macromolecule decomposed by microorganisms was lignin, along with its diagenetic derivatives. The biogas released during the process contained CO2 and trace amounts of CH4 (up to 50 ppm). Isotopic data indicated the occurrence of anaerobic CH4 oxidation. Notably, 16S rRNA gene sequencing identified the presence of N2-fixing microorganisms in all investigated samples, members of the order Rhizobiales (families Beijerinckiaceae, Rhizobiaceae). Our findings demonstrate that N2-fixation may play a pivotal role in coal decomposition under anaerobic conditions.

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Michal Bucha, Piotr Siupka, Anna Detman-Ignatowska, Sushmita Deb, Agnieszka Bylina, Daria Kaim, Aleksandra Chojnacka, Dominika Kufka, Dominika Lewicka-Szczebak, Anna Sikora, and Leszek Marynowski

Status: open (until 17 Mar 2025)

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Michal Bucha, Piotr Siupka, Anna Detman-Ignatowska, Sushmita Deb, Agnieszka Bylina, Daria Kaim, Aleksandra Chojnacka, Dominika Kufka, Dominika Lewicka-Szczebak, Anna Sikora, and Leszek Marynowski
Michal Bucha, Piotr Siupka, Anna Detman-Ignatowska, Sushmita Deb, Agnieszka Bylina, Daria Kaim, Aleksandra Chojnacka, Dominika Kufka, Dominika Lewicka-Szczebak, Anna Sikora, and Leszek Marynowski

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Short summary
In this work, we show the evidences of anaerobic activity of nitrogen-processing microorganisms that naturally occurred in outcropped organic-rich sediments as Miocene lignites. The results presented in this paper indicate the important role of nitrogen supply processes in environments depleted in this element and at the same time rich in humic substances, such as peat bogs, wetlands, immature sediments containing organic carbon (e.g. gyttja, fossil wood, coal).
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