Abstract
Acetogens play a key role in anaerobic degradation of organic material and in maintaining biogas process efficiency. Profiling this community and its temporal changes can help evaluate process stability and function, especially under disturbance/stress conditions, and avoid complete process failure. The formyltetrahydrofolate synthetase (FTHFS) gene can be used as a marker for acetogenic community profiling in diverse environments. In this study, we developed a new high-throughput FTHFS gene sequencing method for acetogenic community profiling and compared it with conventional terminal restriction fragment length polymorphism of the FTHFS gene, 16S rRNA gene-based profiling of the whole bacterial community, and indirect analysis via 16S rRNA profiling of the FTHFS gene-harbouring community. Analyses and method comparisons were made using samples from two laboratory-scale biogas processes, one operated under stable control and one exposed to controlled overloading disturbance. Comparative analysis revealed satisfactory detection of the bacterial community and its changes for all methods, but with some differences in resolution and taxonomic identification. FTHFS gene sequencing was found to be the most suitable and reliable method to study acetogenic communities. These results pave the way for community profiling in various biogas processes and in other environments where the dynamics of acetogenic bacteria have not been well studied.
Highlights
Acetogens play a key role in anaerobic degradation of organic material and in maintaining biogas process efficiency
The aims of the present study were to (1) further evaluate AcetoBase and AcetoScan for profiling and monitoring the temporal dynamics of acetogenic communities in biogas reactors and (2) compare this new highthroughput amplicon sequencing (AmpSeq) method targeting the acetogenic community with conventional methods such as terminal restriction fragment length polymorphism (T-RFLP) and 16S rRNA gene sequencing
The content of carbon dioxide (%) and of methane (%) and volatile fatty acids (VFA) levels were used as indicators of process performance of both GR1 and GR2
Summary
Acetogens play a key role in anaerobic degradation of organic material and in maintaining biogas process efficiency. Less expensive and relatively easier molecular marker-based analysis techniques are generally used to track the temporal dynamics of the whole microbial community involved in the biogas process or a selected fraction of that community. These techniques include fluorescence in-situ hybridisation (FISH), single-strand conformation polymorphism (SSCP), denaturing gradient gel electrophoresis (DGGE), quantitative real-time polymerase chain reaction (qRT-PCR), terminal restriction fragment length polymorphism (T-RFLP) and amplicon sequencing (AmpSeq) of the 16S rRNA gene[28,46]. Our previous studies showed that AcetoBase and AcetoScan can be used for reliable monitoring of the acetogenic community in multiplexed samples in biogas reactors
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