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bioRxiv · 10.1101/2025.10.06.680780

Enhancing Volatile Fatty Acid Accumulation in Seaweed-Arrested Anaerobic Digestion via a Two-Tier Framework of Engineering Diagnostics and Microbial Surveillance

Zhao, Z.·Liu, S.·Nikkhah, H.·Zhang, Y.·Wang, Y.·Liang, J.·Lu, S.·Beykal, B.·Qiao, M.·Li, B.
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Abstract

This study presents a two-tier framework for brown seaweed-arrested anaerobic digestion (SW-AAD) by coupling engineering diagnostics (tier-1) with microbial surveillance (tier-2) to transform conventional digesters into volatile fatty acid (VFAs) biorefineries. In 70-day lab-scale batch tests (volume: 2 L), engineering diagnostics revealed that sequential additions of brown seaweed elevated salinity to 3.3% ash content, stabilized pH at 5.8-6.1, collapsed the CH/CO2 ratio from 2.98 to <0.04, suppressed biogas by 96%, and boosted carbon-conversion efficiency from 10% to 52%. More than half of the influent carbon was redirected from methane to a liquid-phase VFAs pool, dominated by butyrate, hexanoate, and acetate, peaking at 14.5 g L-{superscript 1}. Microbial surveillance using 16S rRNA sequencing presented a 10-fold decline in methanogens, alongside an increase in salt- and acid-tolerant acidogenic microbial families, including Lachnospiraceae, Ruminococcaceae, and Clostridiaceae, as well as seaweed-derived microbial families, including Psychromonadaceae, and Marinomonadaceae. Temporal synchrony and asynchrony-resolved analysis confirmed a strong correlation between two tiers, revealing an operational window featured with two patterns: a rapid engineering parameter response (where changes in engineering parameters were followed by a decline in methanogens within 0-3 days) and a slower microbial restructuring (where the surge in VFA concentration lagged the enrichment of seaweed-derived acidogens by 1-5 days). Based on this operation window, feedback control strategies were simulated for SW-AAD, indicating that cumulative VFAs (cVFAs) yield could be bolstered from 11.38 to 16.13 g COD L-{superscript 1} (+41.8%). A techno-economic assessment (TEA) revealed that this VFA-targeted SW-AAD can reduce capital expenditure by [~]20% compared to conventional AD. This study underscores the promise of asynchrony-resolved analysis of SW-AAD systems for robust and economically viable VFAs production from organic waste. Graphics O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=131 SRC="FIGDIR/small/680780v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@1dc596dorg.highwire.dtl.DTLVardef@1731cadorg.highwire.dtl.DTLVardef@5fe025org.highwire.dtl.DTLVardef@74d8d3_HPS_FORMAT_FIGEXP M_FIG C_FIG Environmental synopsisBrown seaweed-arrested anaerobic digestion (SW-AAD) leverages seaweed-driven salinity, and acidification to suppress methanogenesis, diverting >50% of carbon to VFAs (14.5 g COD L-{superscript 1}) while cutting methane by 99%. A two-tier framework couples engineering diagnostics with microbial surveillance and asynchrony-resolved analysis, enabling feedback control and low-emission waste valorization in digesters.

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Zhao, Z., Liu, S., Nikkhah, H., Zhang, Y., Wang, Y., Liang, J., Lu, S., Beykal, B., Qiao, M., Li, B.. 2025-10-07. Enhancing Volatile Fatty Acid Accumulation in Seaweed-Arrested Anaerobic Digestion via a Two-Tier Framework of Engineering Diagnostics and Microbial Surveillance. https://doi.org/10.1101/2025.10.06.680780

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