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bioRxiv · 10.1101/2025.10.15.682543

Microbial community analyses of composts are influenced by particle size fraction and DNA extraction method

Logo, A.·Koch, T.·Maurhofer, M.·Oberhaensli, T.·Thuerig, B.·Widmer, F.·Flury, P.·Mayerhofer, J.
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Abstract

Composting plays a key role in sustainable agriculture by converting organic waste into a valuable soil conditioner. The process is driven by complex microbial communities, whose characterization is essential for optimizing the composting process and compost quality. Molecular techniques such as amplicon sequencing are commonly used for this purpose. However, sampling procedures and DNA extraction methods, key steps in the sequencing workflow, vary often across studies, challenging comparability. We investigated two aspects of sampling preparation that may influence compost microbial analyses. For DNA extraction, often fine fractions (<2 mm) are used. However, compost has a heterogeneous structure, including coarse particles. To assess the effect of particle size, we separately sequenced bacterial and fungal communities of the fine (0-2 mm) and coarse (2-10 mm) fractions of three composts. In addition, DNA was extracted using a carboxyl affinity-based magnetic method and a silanol affinity-based filter method to evaluate the impact of the extraction technique. We found that the coarse fraction had higher bacterial richness and a distinct bacterial and fungal community structure compared to the fine fraction. DNA extraction method also influenced bacterial community profiles, with the magnetic bead method improving coverage, particularly for Bacillota. Although the effects of particle size and extraction method were small compared to general diversity among composts, we recommend including coarse particles in sequencing analyses and using standardized DNA extraction protocols, especially for studies aiming at high-resolution community analyses. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/682543v1_ufig1.gif" ALT="Figure 1"> View larger version (51K): org.highwire.dtl.DTLVardef@1cd1b77org.highwire.dtl.DTLVardef@c64c9corg.highwire.dtl.DTLVardef@cfb5f6org.highwire.dtl.DTLVardef@b350df_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Logo, A., Koch, T., Maurhofer, M., Oberhaensli, T., Thuerig, B., Widmer, F., Flury, P., Mayerhofer, J.. 2025-10-15. Microbial community analyses of composts are influenced by particle size fraction and DNA extraction method. https://doi.org/10.1101/2025.10.15.682543

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