High diversity and abundance of putative polyphosphate-accumulating Tetrasphaera-related bacteria in activated sludge systems

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Project Description

Investigated Tetrasphaera bacteria involved in phosphorus removal in wastewater treatment.
Found to be diverse, comprising up to ~30% of the microbial community.
Contributes to phosphorus removal by storing it as polyphosphate.
Activated after consuming specific carbon sources (e.g., amino acids, glucose) under anaerobic conditions.
Complements other bacteria, enhancing system stability and overall performance.

A total of 23,264 16S rRNA gene clone sequences were analyzed from EBPR (Enhanced Biological Phosphorus Removal) systems.
Tetrasphaera-related bacteria were the most abundant group, accounting for 11.6% of total sequences.
Phylogenetic analysis using maximum likelihood identified several distinct clades within the Tetrasphaera lineage.
New FISH probes were designed for major Tetrasphaera groups and applied successfully in full-scale EBPR plants.
Microscopy revealed diverse morphologies, including rods, tetrads, and filaments, for Tetrasphaera cells.
In situ quantification showed Tetrasphaera were often more abundant than ‘Candidatus Accumulibacter’, the traditionally recognized PAO.
MAR-FISH analysis confirmed that Tetrasphaera can:
- Uptake and store polyphosphate (poly-P).
- Ferment glucose under anaerobic conditions.
The findings suggest that Tetrasphaera plays a central role in phosphorus removal in EBPR systems—more so than previously recognized.

The study successfully achieved its initial objectives:

Clarified the genetic, morphological, and phylogenetic diversity of Tetrasphaera in full-scale EBPR systems.
Confirmed their key role in EBPR, including their ability to accumulate polyphosphate under specific conditions.
Revealed their high abundance, often exceeding that of Accumulibacter, in multiple WWTPs.
Expanded understanding of their ecophysiology, demonstrating metabolic traits distinct from classical PAOs.
Provided practical insights for microbial monitoring and potential manipulation of PAO communities to enhance EBPR performance.

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