Delafont, Vincent

ABSTRACT Ca . Babelota is a phylum of strictly intracellular bacteria whose representatives are commonly detected in various environments through metagenomics, though their presence, ecology, and biology have never been addressed so far. As a group of strict intracellular, we hypothesize that their presence, occurrence, and abundance heavily depend on their hosts, which are known as heterotrophic protists, based on few described isolates. Here, we conducted a sampling campaign allowing to characterize protists and associated bacterial communities, using high-throughput sequencing. In parallel, a systematic enrichment of protists from samples was performed to attempt characterization and isolation of new Ca . Babelota within native hosts. We found that Ca . Babelota are among the most widespread phylum among the rare ones. Protist enrichments are allowed in certain cases to enrich as well for Ca . Babelota, which could be visualized in vivo infecting protist cells. Though cosmopolitan, Ca . Babelota diversity was highly site-specific. Cooccurrence analyses allowed to retrieve well-known as well as new putative associations involving numerous protists of various trophic regimes. The combination of approaches developed in this study enhances our understanding of Ca . Babelota ecology and biology, while paving the way for future isolation of new members of this elusive phylum, which could have huge impact on protists—and ecosystems—functioning. IMPORTANCE Our understanding of microbial diversity surrounding us and colonizing the environment has been dramatically impacted by the advent of DNA-based analyses. Such progress helped shine a new light on numerous lineages of yet-to-be-characterized microbes, whose ecology and biology are basically unknown. Among those uncharacterized clades is the Candidatus Babelota, a bacterial phylum for which parasitism seems to be an ancestral trait. All known Ca. Babelota thrive by infecting phagotrophic protist hosts, thereby impacting this basal link of the trophic chain. The Ca. Babelota constitutes a model that stands out, as phylum-wide conserved parasitism has only been described in one previous occurrence for Bacteria, with the Chlamydiota. Thus, exploring the intricate interplay between Ca. Babelota and their protist hosts will advance our knowledge of bacterial diversity, their ecology, and global impact on ecosystem functioning.

Abstract Ba.be.lo'ta. N.L. fem. n. Babela , a Candidatus genus name; L. neut. pl. n. suff. ‐ ota , ending to denote a phylum; N.L. neut. pl. n. Babelota , the Candidatus Babela phylum. Candidatus Babelota is a phylum of strictly intracellular bacteria, currently divided into two class‐level groups and at least five orders based on 16S rRNA sequence analyses. However, all known biological isolates to date belong to the same order of the Babelales, within the Babelia class. Presently, four isolates representing three families have been described and maintained in laboratory cultures through the cultivation of their hosts, which are known to be amoeboid and flagellated protists. Genomic analysis of Ca . Babelota consistently reveals a profound reduction in metabolic capabilities, suggesting a phylum‐wide, conserved parasitic lifestyle. To date, all lineages covered by genomic data indicate that parasitism is a defining trait of this phylum. Ca . Babelota exhibits a cosmopolitan environmental distribution, with sequences detected across a spectrum of habitats including soil, freshwater, and saline or hypersaline water bodies. Taxonomic and Nomenclature Notes According to the List of Prokaryotic names with Standing in Nomenclature (LPSN), the taxonomic status of the phylum Candidatus Babelota is: preferred name (not correct name) (last update, February 2025) * . LPSN classification: Bacteria / incertae sedis / Candidatus Babelota The phylum Candidatus Babelota can also be recovered in the Genome Taxonomy Database (GTDB) as p__Dependentiae (version v220) ** . GTDB classification: d__Bacteria / p__Dependentiae * Meier‐Kolthoff et al. ( 2022 ). Nucleic Acids Res , 50 , D801 – D807 ; DOI: 10.1093/nar/gkab902 ** Parks et al. ( 2022 ). Nucleic Acids Res , 50 , D785 – D794 ; DOI: 10.1093/nar/gkab776