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It takes two to tango without O2: Metabolic interactions between breviate protists and bacteria in anoxia

Aguilera Campos, Karla Iveth LU orcid (2026)
Abstract
Breviates are anaerobic protists and among the closest unicellular relatives of animals and fungi, yet their ecology and metabolism remain poorly understood. In this thesis, I investigated the distribution, metabolic potential, and bacterial interactions of breviates in low-oxygen environments. Breviates rely on bacteria both as a food source and potentially as metabolic partners. Breviates are predicted to produce hydrogen gas via [FeFe]-hydrogenases, whose activity requires low hydrogen partial pressure; therefore, hydrogen-consuming partners are essential to sustain breviate metabolism.

Using metagenomics, 16 high-quality bacterial genomes were reconstructed from protist-associated Arcobacteraceae, Desulfovibrionaceae, and... (More)
Breviates are anaerobic protists and among the closest unicellular relatives of animals and fungi, yet their ecology and metabolism remain poorly understood. In this thesis, I investigated the distribution, metabolic potential, and bacterial interactions of breviates in low-oxygen environments. Breviates rely on bacteria both as a food source and potentially as metabolic partners. Breviates are predicted to produce hydrogen gas via [FeFe]-hydrogenases, whose activity requires low hydrogen partial pressure; therefore, hydrogen-consuming partners are essential to sustain breviate metabolism.

Using metagenomics, 16 high-quality bacterial genomes were reconstructed from protist-associated Arcobacteraceae, Desulfovibrionaceae, and Terasakiellaceae, revealing one new genus and nine new species. All bacterial genomes encoded [NiFe]-hydrogenases, indicating hydrogen uptake potential, long with diverse sulfur and nitrogen metabolic pathways. The relative abundance of associated bacteria shifted with the availability of electron acceptors, with nitrate reducers being more abundant in the presence of nitrate and sulfate reducers in the presence of sulfate.

Growth experiments showed that breviates grew well in the presence of nitrate when hydrogenotrophic denitrifying bacteria were present, as shown before for the breviate Lenisia limosa. In addition, growth experiments showed that Pygsuia biforma reached significantly higher cell densities when co-cultured with a sulfate-reducing bacterium (SRB) under sulfate-replete conditions than under sulfate-free conditions or without SRB. Transcriptomic analyses revealed that sulfate deprivation induced oxidative stress responses in P. biforma, whereas sulfate availability supported a more active metabolic state.

Together, these results support a syntrophic model in which SRB perform dissimilatory sulfate reduction using breviate-derived hydrogen as an electron donor, while the breviates use the resulting sulfide for Fe–S cluster and amino acid biosynthesis. This work advances our understanding of how breviate protists interact with bacteria in low-oxygen environments and how these interactions influence the hydrogen, nitrogen, and sulfur cycles (Less)
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author
supervisor
opponent
  • Associate Professor Marshall, Ian, Department of Biology, Aarhus University.
organization
publishing date
type
Thesis
publication status
published
subject
keywords
metabolic interactions, syntrophy, bacteria, protists, breviate, anoxia, Arcobacteraceae, Desulfovibrionaceae, Terasakiella, sulfate, nitrate, hydrogen
pages
88 pages
publisher
Lund
defense location
Hörsal A213 Hall, Department of Biology, Sölvegatan 35, Lund, Sweden.
defense date
2026-02-26 13:00:00
ISBN
978-91-8104-773-8
978-91-8104-774-5
language
English
LU publication?
yes
id
13c6f6f6-b8fc-46a7-9b42-bb4baca7f64b
date added to LUP
2026-01-26 11:06:03
date last changed
2026-01-27 13:37:38
@phdthesis{13c6f6f6-b8fc-46a7-9b42-bb4baca7f64b,
  abstract     = {{Breviates are anaerobic protists and among the closest unicellular relatives of animals and fungi, yet their ecology and metabolism remain poorly understood. In this thesis, I investigated the distribution, metabolic potential, and bacterial interactions of breviates in low-oxygen environments. Breviates rely on bacteria both as a food source and potentially as metabolic partners. Breviates are predicted to produce hydrogen gas via [FeFe]-hydrogenases, whose activity requires low hydrogen partial pressure; therefore, hydrogen-consuming partners are essential to sustain breviate metabolism. <br/><br/>Using metagenomics, 16 high-quality bacterial genomes were reconstructed from protist-associated Arcobacteraceae, Desulfovibrionaceae, and Terasakiellaceae, revealing one new genus and nine new species. All bacterial genomes encoded [NiFe]-hydrogenases, indicating hydrogen uptake potential, long with diverse sulfur and nitrogen metabolic pathways. The relative abundance of associated bacteria shifted with the availability of electron acceptors, with nitrate reducers being more abundant in the presence of nitrate and sulfate reducers in the presence of sulfate. <br/><br/>Growth experiments showed that breviates grew well in the presence of nitrate when hydrogenotrophic denitrifying bacteria were present, as shown before for the breviate Lenisia limosa. In addition, growth experiments showed that Pygsuia biforma reached significantly higher cell densities when co-cultured  with a sulfate-reducing bacterium (SRB) under sulfate-replete conditions than under sulfate-free conditions or without SRB. Transcriptomic analyses revealed that sulfate deprivation induced oxidative stress responses in P. biforma, whereas sulfate availability supported a more active metabolic state. <br/><br/>Together, these results support a syntrophic model in which SRB perform dissimilatory sulfate reduction using breviate-derived hydrogen as an electron donor, while the breviates use the resulting sulfide for Fe–S cluster and amino acid biosynthesis. This work advances our understanding of how breviate protists interact with bacteria in low-oxygen environments and how these interactions influence the hydrogen, nitrogen, and sulfur cycles}},
  author       = {{Aguilera Campos, Karla Iveth}},
  isbn         = {{978-91-8104-773-8}},
  keywords     = {{metabolic interactions; syntrophy; bacteria; protists; breviate; anoxia; Arcobacteraceae; Desulfovibrionaceae; Terasakiella; sulfate; nitrate; hydrogen}},
  language     = {{eng}},
  month        = {{01}},
  publisher    = {{Lund}},
  school       = {{Lund University}},
  title        = {{It takes two to tango without O2: Metabolic interactions between breviate protists and bacteria in anoxia}},
  year         = {{2026}},
}