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case-control·gastroenterology, infectious disease, clinical trial, research methods·PMC10608849

Ethanol-Producing Enterocloster bolteae Is Enriched in Chronic Hepatitis B-Associated Gut Dysbiosis: A Case–Control Culturomics Study

Microorganisms · 10 authors, 4 centres

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TL;DR

This case–control study compared gut microbiota in chronic HBV patients versus healthy controls using both culturomics and 16S rDNA metagenomic sequencing.

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This case–control study compared gut microbiota in chronic HBV patients versus healthy controls using both culturomics and 16S rDNA metagenomic sequencing. Among 18 total samples, culturomics identified 240 species from 14,340 colonies in the HBV group and 254 species from 14,780 colonies in controls. Enterocloster and Clostridium were significantly enriched by culturomics in HBV patients, while 213 species were missing exclusively from the HBV group. Limitations include the small sample size and the study's exploratory design, and further studies with larger cohorts are encouraged to confirm these findings.

STUDY TYPE

case-control

SPECIALTY

gastroenterology

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100% · A

ORIGINAL PAPER

PMC10608849

Ethanol-Producing Enterocloster bolteae Is Enriched in Chronic Hepatitis B-Associated Gut Dysbiosis: A Case–Control Culturomics Study

Microorganisms

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ABSTRACT

Abstract Background: Hepatitis B virus (HBV) infection is a global health epidemic that causes fatal complications, leading to liver cirrhosis and hepatocellular carcinoma. The link between HBV-related dysbiosis and specific bacterial taxa is still under investigation. Enterocloster is emerging as a new genus (formerly Clostridium ), including Enterocloster bolteae , a gut pathogen previously associated with dysbiosis and human diseases such as autism, multiple sclerosis, and inflammatory bowel diseases. Its role in liver diseases, especially HBV infection, is not reported. Methods: The fecal samples of eight patients with chronic HBV infection and ten healthy individuals were analyzed using the high-throughput culturomics approach and compared to 16S rRNA sequencing. Quantification of ethanol, known for its damaging effect on the liver, produced from bacterial strains enriched in chronic HBV was carried out by gas chromatography–mass spectrometry. Results: Using culturomics, 29,120 isolated colonies were analyzed by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI–TOF); 340 species were identified (240 species in chronic HBV samples, 254 species in control samples) belonging to 169 genera and 6 phyla. In the chronic HBV group, 65 species were already known in the literature; 48 were associated with humans but had not been previously found in the gut, and 17 had never been associated with humans previously. Six species were newly isolated in our study. By comparing bacterial species frequency, three bacterial genera were serendipitously found with significantly enriched bacterial diversity in patients with chronic HBV: Enterocloster , Clostridium , and Streptococcus ( p = 0.0016, p = 0.041, p = 0.053, respectively). However, metagenomics could not identify this enrichment, possibly concerning its insufficient taxonomical resolution (equivocal assignment of operational taxonomic units). At the species level, the significantly enriched species in the chronic HBV group almost all belonged to class Clostridia , such as Clostridium perfringens , Clostridium sporogenes , Enterocloster aldenensis , Enterocloster bolteae , Enterocloster clostridioformis , and Clostridium innocuum. Two E. bolteae strains, isolated from two patients with chronic HBV infection, showed high ethanol production (27 and 200 mM). Conclusions: Culturomics allowed us to identify Enterocloster species, specifically, E. bolteae , enriched in the gut microbiota of patients with chronic HBV. These species had never been isolated in chronic HBV infection before. Moreover, ethanol production by E. bolteae strains isolated from the chronic HBV group could contribute to liver disease progression. Additionally, culturomics might be critical for better elucidating the relationship between dysbiosis and chronic HBV infection in the future. Keywords: dysbiosis, gut microbiota, chronic hepatitis B virus infection, Enterocloster bolteae , culturomics, metagenomics

CITATION EXPORT

Reham Magdy Wasfy, Babacar Mbaye, Patrick Borentain et al.. Ethanol-Producing Enterocloster bolteae Is Enriched in Chronic Hepatitis B-Associated Gut Dysbiosis: A Case–Control Culturomics Study. Microorganisms. (2023). https://doi.org/10.3390/microorganisms11102437

AUTHORS

10 · 4 CENTRES

Reham Magdy Wasfy, Babacar Mbaye, Patrick Borentain, Maryam Tidjani Alou, Maria Leticia Murillo Ruiz, Aurelia Caputo, Claudia Andrieu, Nicholas Armstrong, Matthieu Million, Rene Gerolami

1IHU Méditerranée Infection, 13005 Marseille, [email protected] (M.T.A.); [email protected] (C.A.);
2MEPHI, IRD, Aix-Marseille Université, 13005 Marseille, France
3Unité Hépatologie, Hôpital de la Timone, APHM, 13005 Marseille, France; [email protected]
4Assistance Publique-Hôpitaux de Marseille (APHM), 13005 Marseille, France