Search bioRxiv⌕ Search
Home/Archive/bioRxiv

bioRxiv · 10.1101/696351

Long solids retention times and attached growth phase favor prevalence of comammox bacteria in nitrogen removal systems.

Cotto, I.·Dai, Z.·Huo, L.·Anderson, C. L.·Vilardi, K. J.·Ijaz, U. Z.·Khunjar, W.·Wilson, C.·De Clippeleir, H.·Gilmore, K.·Bailey, E.·Pinto, A.
View original source ↗Export metadata ↓

Abstract

The discovery of the complete ammonia oxidizing (comammox) bacteria overturns the traditional two-organism nitrification paradigm which largely underpins the design and operation of nitrogen removal during wastewater treatment. Quantifying the abundance, diversity, and activity of comammox bacteria in wastewater treatment systems is important for ensuring a clear understanding of the nitrogen biotransformations responsible for ammonia removal. To this end, we conducted a yearlong survey of 14 full-scale nitrogen removal systems including mainstream conventional and simultaneous nitrification-denitrification and side-stream partial nitrification-anammox systems with varying process configurations. Metagenomics and genome-resolved metagenomics identified comammox bacteria in mainstream conventional and simultaneous nitrification-denitrification systems, with no evidence for their presence in side-stream partial nitrification-anammox systems. Further, comammox bacterial diversity was restricted to clade A and these clade A comammox bacteria were detected in systems with long solids retention times (>10 days) and/or in the attached growth phase. Using a newly designed qPCR assay targeting the amoB gene of clade A comammox bacteria in combination with quantitation of other canonical nitrifiers, we show that long solids retention time is the key process parameter associated with the prevalence and abundance of comammox bacteria. The increase in comammox bacterial abundance was not associated with concomitant decrease in the abundance of canonical nitrifiers; however, systems with comammox bacteria showed significantly better and temporally stable ammonia removal compared to systems where they were not detected. Finally, in contrast to recent studies, we do not find any significant association of comammox bacterial prevalence and abundance with dissolved oxygen concentrations in this study.\n\nHighlightsO_LIClade A comammox bacteria were detected in wastewater nitrogen removal systems.\nC_LIO_LINew qPCR assay targeting the amoB gene of clade A comammox bacteria was developed.\nC_LIO_LIComammox bacteria are prevalent in mainstream conventional and simultaneous nitrification-denitrification systems with long solids retention times (>10 days).\nC_LIO_LIComammox bacteria were not detected in sidestream partial nitrification-anammox systems included in this study.\nC_LI\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=127 SRC=\"FIGDIR/small/696351v2_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (28K):\norg.highwire.dtl.DTLVardef@3228c9org.highwire.dtl.DTLVardef@113dfb8org.highwire.dtl.DTLVardef@1c4adaforg.highwire.dtl.DTLVardef@166f997_HPS_FORMAT_FIGEXP M_FIG GRAPHICAL ABSTRACT\n\nC_FIG

Source connections

Explore related subjects

microbiologyPreprint

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Cotto, I., Dai, Z., Huo, L., Anderson, C. L., Vilardi, K. J., Ijaz, U. Z., Khunjar, W., Wilson, C., De Clippeleir, H., Gilmore, K., Bailey, E., Pinto, A.. 2019-07-09. Long solids retention times and attached growth phase favor prevalence of comammox bacteria in nitrogen removal systems.. https://doi.org/10.1101/696351

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Polyphyletic origins and lineage-specific expansion of chemosensory systems in phylum Actinomycetota

Phylum Actinomycetota (syn. Actinobacteriota or Actinobacteria), one of the largest well-studied bacterial phyla, is renowned for complex morphology, non-motility, ecological versatility, and secondary metabolite production. Chemosensory systems (CSS) are among the most sophisticated prokaryotic signal transduction architectures, yet their evolutionary distribution in this phylum remains poorly understood. Here, a comparative genomic analysis of 693 high-quality genomes spanning the phylum resolves the diversity, distribution, and evolutionary trajectories of CSS. Despite extensive genomic complexity and ecological versatility, only 6.9% of genomes encode complete CSS clusters, confined to a subset of lineages within a few classes, indicating lineage-specific retention or loss. In contrast, two-component systems (TCS) are ubiquitously abundant, with hybrid kinases enriched in CSS-deficient classes, suggesting functional compensation for the absence of dedicated chemosensory pathways. Basal lineages retain simplified CSS (F1 or F5), whereas Actinomycetes show lineage-specific expansion into multiple classes (F5, F8, F9, and ACF), often co-occurring, indicative of enhanced sensory complexity. The phylogeny of CheA homologs shows a polyphyletic distribution, supporting a mosaic evolutionary history shaped by multiple independent acquisitions, diversification, and gene loss. Collectively, our findings show that, unlike motile bacteria, members of Actinomycetota rely predominantly on TCS-driven regulatory networks, with CSS retained only in select lineages where ecological demands require enhanced environmental sensing. This work provides a phylum-wide evolutionary framework for the emergence, diversification, and functional specialization of CSS in non-motile bacteria.

microbiology↗
bioRxiv2026

Integrating Microbial Ecology and Sustainability: Linking Agricultural Waste Gas Streams to Microbial Community Shifts in a Photobioreactor

Photobioreactors (PBRs) coupled to animal housing offer potential for mitigating gaseous emissions such as CO2 and NH3 while simultaneously producing biomass. Here, we present a time-resolved characterization of microbial community dynamics, with a focus on the genetic potential for carbon and nitrogen metabolism. Using a cone-shaped, helical PBR cultivating Arthrospira platensis (Spirulina) and supplied with exhaust air from a chicken house, we characterized microbial communities over continuous exhaust-air exposure and periodic biomass harvesting. Microbiome and gene-centric profiling revealed temporal variation in community composition and in the representation of genes associated with carbon and nitrogen metabolism. Cycle-based dissolved inorganic nitrogen balances showed that the decrease in dissolved inorganic nitrogen was only partially recovered in harvested biomass, while ammonia-oxidizing microorganisms were associated with structured, low-light niches. These observations indicate ammonia-oxidizing potential but do not establish a quantitatively relevant nitrification flux. Structured carrier matrices provided attached-growth niches harboring microbial populations underrepresented in the circulating suspension, while pre-colonized carriers facilitated biomass development during re-inoculation. Overall, our results show that coupling PBRs to agricultural exhaust streams can support biomass production while sustaining spatially structured microbial communities with taxonomic and functional potential. These findings may inform their further development for emission mitigation and circular biomass production.

microbiology↗
bioRxiv2026

Engineered DNA materials shape microbial physiology and ecological selection

Extracellular DNA (eDNA) is both a structural component of microbial biofilms and a potential reservoir of genetic information, yet its role as an ecological material remains largely unexplored. Here we establish engineered extracellular materials as determinants of microbial physiology and community assembly across increasing biological complexity. A three-dimensional DNA hydrogel altered bacterial stress-associated responses, elicited species-dependent phenotypes and transcriptional changes in a defined consortium, and shaped community succession in wastewater-derived microbiomes. Distinct responses to surface-bound oligonucleotides reveal that DNA presentation matters beyond its mere presence. Strikingly, metagenomic sequencing identified rare, hydrogel-specific candidate junctions linking material-derived DNA to microbial genomic sequences, suggesting a previously unexplored interface between extracellular DNA materials and microbial genetic information. These findings reveal a dual role for engineered DNA: its material organization shapes microbial physiology and ecological selection, while its encoded sequences may engage with microbial genomic processes.

microbiology↗