Khoudia Diop

Culture of previously uncultured members of the human gut microbiota by culturomics

Metagenomics revolutionized the understanding of the relations among the human microbiome, health and diseases, but generated a countless number of sequences that have not been assigned to a known microorganism1. The pure culture of prokaryotes, neglected in recent decades, remains essential to elucidating the role of these organisms2. We recently introduced microbial culturomics, a culturing approach that uses multiple culture conditions and matrix-assisted laser desorption/ionization–time of flight and 16S rRNA for identification2. Here, we have selected the best culture conditions to increase the number of studied samples and have applied new protocols (fresh-sample inoculation; detection of microcolonies and specific cultures of Proteobacteria and microaerophilic and halophilic prokaryotes) to address the weaknesses of the previous studies3–5. We identified 1,057 prokaryotic species, thereby adding 531 species to the human gut repertoire: 146 bacteria known in humans but not in the gut, 187 bacteria and 1 archaea not previously isolated in humans, and 197 potentially new species. Genome sequencing was performed on the new species. By comparing the results of the metagenomic and culturomic analyses, we show that the use of culturomics allows the culture of organisms corresponding to sequences previously not assigned. Altogether, culturomics doubles the number of species isolated at least once from the human gut.

‘Arcanobacterium urinimassiliense’ sp. nov., a new bacterium isolated from the urogenital tract

Herein we report the main characteristics of ‘Arcanobacterium urinimassiliense’ strain Marseille-P3248T (=CSUR P3248) isolated from a urine sample of a 54-day-old girl with rotavirus gastroenteritis.

Olegusella massiliensis gen. nov., sp. nov., strain KHD7T, a new bacterial genus isolated from the female genital tract of a patient with bacterial vaginosis

Strain KHD7T, a Gram-stain-positive rod-shaped, non-sporulating, strictly anaerobic bacterium, was isolated from the vaginal swab of a woman with bacterial vaginosis. We studied its phenotypic characteristics and sequenced its complete genome. The major fatty acids were C16:0 (44%), C18:2n6 (22%), and C18:1n9 (14%). The 1,806,744 bp long genome exhibited 49.24% G+C content; 1549 protein-coding and 51 RNA genes. Strain KHD7T exhibited a 93.5% 16S rRNA similarity with Olsenella uli, the phylogenetically closest species in the family Coriobacteriaceae. Therefore, strain KHD7T is sufficiently distinct to represent a new genus, for which we propose the name Olegusella massiliensis gen. nov., sp. nov. The type strain is KHD7T.

Taxonogenomics and description of Vaginella massiliensis gen. nov., sp. nov., strain Marseille P2517T, a new bacterial genus isolated from the human vagina

An obligate aerobic, Gram-negative, nonmotile and nonsporulating rod designated Marseille P2517 was isolated from the vaginal flora. We describe its features, annotate the genome and compare it to the closest species. The 16S rRNA analysis shows 93.03% sequence similarity with Weeksella virosa, the phylogenetically closest species. Its genome is 2 434 475 bp long and presents 38.16% G+C. On the basis of these data, it can be considered as a new genus in the Flavobacteriaceae family, for which we proposed the name Vaginella massiliensis gen. nov., sp. nov. The type strain is Marseille P2517T.

Description of three new Peptoniphilus species cultured in the vaginal fluid of a woman diagnosed with bacterial vaginosis: Peptoniphilus pacaensis sp. nov., Peptoniphilus raoultii sp. nov., and Peptoniphilus vaginalis sp. nov.

Three previously unidentified Gram‐positive anaerobic coccoid bacteria, strains KhD‐2T, KHD4T, and Kh‐D5T, isolated from a vaginal swab, were characterized using the taxonogenomics concept. The phylogenic analysis, phenotypic characteristics, and genotypic data presented in this report attest that these three bacteria are distinct from previously known bacterial species with standing in nomenclature and represent three new Peptoniphilus species. Strain KhD‐2T is most closely related to Peptoniphilus sp. DNF00840 and Peptoniphilus harei (99.7% and 98.2% identity, respectively); strain KHD4T to Peptoniphilus lacrimalis (96%) and strain Kh‐D5T to Peptoniphilus coxii (97.2%). Strains KhD‐2T, KHD4T, and Kh‐D5T DNA G+C contents are, respectively, 34.23%, 31.87%, and 49.38%; their major fatty acid was C16:0 (41.6%, 32.0%, and 36.4%, respectively). We propose that strains KhD‐2T (=CSUR P0125 = DSM 101742), KHD4T (=CSUR P0110 = CECT 9308), and Kh‐D5T (=CSUR P2271 = DSM 101839) be the type strains of the new species for which the names Peptoniphilus vaginalis sp. nov., Peptoniphilus raoultii sp. nov., and Peptoniphilu pacaensis sp. nov., are proposed, respectively.

Characterization of a novel Gram‐stain‐positive anaerobic coccus isolated from the female genital tract: Genome sequence and description of Murdochiella vaginalis sp. nov.

Strain Marseille‐P2341T, a nonmotile, nonspore‐forming, Gram‐stain‐positive anaerobic coccus, was isolated in the vaginal specimen of a patient with bacterial vaginosis using culturomics. Its growth occurred at temperatures ranging from 25 to 42°C, with pH between 6.5 and 8.5, and at NaCl concentrations lower than 5%. The major fatty acids were C18:1n9 (27.7%) and C16:0 (24.4%). Its genome is 1,671,491 bp long with 49.48 mol% of G+C content. It is composed of 1,501 genes: 1,446 were protein‐coding genes and 55 were RNAs. Strain Marseille‐P2341T shared 97.3% of 16S rRNA gene sequence similarity with Murdochiella asaccharolytica, the phylogenetically closest species. These results enabled the classification of strain Marseille‐P2341T as a new species of the genus Murdochiella for which we proposed the name Murdochiella vaginalis sp. nov. The type strain is strain Marseille‐P2341T (=DSM 102237, =CSUR P2341).

Draft Genome Sequence of Ezakiella peruensis Strain M6.X2, a Human Gut Gram-Positive Anaerobic Coccus

ABSTRACT We report here the draft genome sequence of Ezakiella peruensis strain M6.X2T. The draft genome is 1,672,788 bp long and harbors 1,589 predicted protein-encoding genes, including 26 antibiotic resistance genes with 1 gene encoding vancomycin resistance. The genome also exhibits 1 clustered regularly interspaced short palindromic repeat region and 333 genes acquired by horizontal gene transfer.

‘Corynebacterium fournierii,’ a new bacterial species isolated from the vaginal sample of a patient with bacterial vaginosis

Here we describe briefly ‘Corynebacterium fournierii’ strain Marseille P2948 (= CSUR P2948 = DSM103271), a new bacterium that was isolated from the vaginal sample of a 21-year-old woman with bacterial vaginosis.

'Olegusella massiliensis' strain KHD7, a new bacterial genus isolated from the female genital tract

We report the main characteristics of ‘Olegusella massiliensis’ gen. nov., sp. nov., strain KHD7 (= CSUR P2268 = DSM 101849), a new member of the Coriobacteriaceae family isolated from the vaginal flora of a patient with bacterial vaginosis.

‘Peptoniphilus pacaensis’ sp. nov., a new species isolated from human female genital tract

We relate here the primary characteristics of ‘Peptoniphilus pacaensis’ strain KHD5 (= CSUR P2271), a new member of the Peptoniphilus genus. Strain KHD5 was isolated from a vaginal sample of a 33-year-old woman exhibiting a bacterial vaginosis.