Limosilactobacillus fermentum, Lactococcus lactis and Thomasclavelia ramosa are enriched and Methanobrevibacter smithii is depleted in patients with non-alcoholic steatohepatitis

Mbaye, Babacar; Wasfy, Reham Magdy; Alou, Maryam Tidjani; Borentain, Patrick; Andrieu, Claudia; Caputo, Aurélia; Raoult, Didier; Gerolami, René; Million, Matthieu

doi:10.1016/j.micpath.2023.106160

Cited by 10 publications

(9 citation statements)

References 57 publications

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“…Speci cally, T. ramosa, is also increased in NASH patients using different methods. The association of T. ramosa and L. fermentum with NASH con rms previous ndings [10,13,43]. However, this study is the rst to demonstrate a plausible association of E. bolteae with NASH.…”

Section: Discussionsupporting

confidence: 90%

“…These ve species included two members of the recently reclassi ed Limosilactobacillus genus [28]: L.fementum and L.panis. L. fermentum, a known ethanol producer [29], which was found enriched in both culture and sequencing and has previously been associated with ethanol production in NASH [10,13], whereas L. panis was only detected in the 16S amplicon sequencing in a multi-assigned OTU [30]. S. mutans, the only member of the genus Streptococcus able to produce ethanol [31], was found nonequivocally using amplicon sequencing (Fig.…”

Section: Comparison Of Culturomics and Amplicon Sequencing Resultsmentioning

confidence: 82%

“…Zhu et al found high concentrations of ethanol in the blood of NASH patients, con rmed in patients' stools by Yuan et al [8,9]. Case-control studies have shown the involvement of speci c microorganisms in endogenous ethanol production, notably yeasts of the Candida and Pichia genus [12] and bacteria including Klebsiella pneumoniae, Limosilactobacillus fermentum and Lactococcus lactis [9,13].…”

Section: Introductionmentioning

confidence: 92%

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The Ethanol-Producing Enterocloster bolteae and Limosilactobacillus fermentum are enriched in Nonalcoholic steatohepatitis

MBAYE,

WASFY,

BORENTAIN

et al. 2023

Preprint

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Background: Non-alcoholic steatohepatitis (NASH) has become a major public health issue. In fact, it is one of the leading causes of disturbed liver function and liver transplantation worldwide. Its link with the intestinal microbiota is still topical, and the production of endogenous ethanol following dysbiosis of the microbiota - one of the mechanisms linked to the disease - has been shown in several studies. In this study, we analyzed stool samples from 41 NASH patients and 24 healthy controls using culturomics and 16S rRNA targeted metagenomics to identify the microbial profile associated with each group. Fecal ethanol concentration was also determined for all samples. Results: We identified 358 different bacterial species using the culturomics approach distributed into 11 phyla and 143 genera. With a significant difference, 12 appeared significantly more frequently in the NASH group and 4 more frequently in the control group. Finegoldia magna was the most detected in NASH patients (10/14 (71%)), and 2 ethanol producing strains: Limosilactobacillus fermentum and Enterocloster bolteae. Metagenomics shows an enrichment of Lactobacillus and Streptococcus and a microbial profile identical to that of the culture. With regard to the species, there was an elevated frequency of ethanol-producing bacteria in NASH patients, and the concentration of ethanol was also higher in the stools of NASH patients compared to healthy controls (bilateral Mann-Whitney test, p=value 0.0145). However, compared with metagenomics, only the culture showed enrichment of Enterocloster bolteae in NASH patients. Conclusions: Culturomics and metagenomics are complementary methods for exploring the microbiota. Our results suggest that enrichment of ethanol-producing bacteria in NASH patients may play an instrumental role in the pathogenesis of the disease.

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Section: Discussionsupporting

confidence: 90%

Section: Comparison Of Culturomics and Amplicon Sequencing Resultsmentioning

confidence: 82%

Section: Introductionmentioning

confidence: 92%

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The Ethanol-Producing Enterocloster bolteae and Limosilactobacillus fermentum are enriched in Nonalcoholic steatohepatitis

MBAYE,

WASFY,

BORENTAIN

et al. 2023

Preprint

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“…Thus, only the first 24 samples (14 NASH and 10 controls) were analyzed using culturomics whereas all samples (41 NASH and 24 controls) were analyzed using V3V4 16S amplicon sequencing simultaneously. No sample was common to our previous studies ( Mbaye et al., 2022 ; Mbaye et al., 2023 ) allowing us to test reproducibility of our previous findings using different methods (specific yeast and enterobacterial microbial culture were used in our previous study but not untargeted culturomics). Characteristics of the population in this study are described in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

“…( Zhu et al., 2013 ; Yuan et al., 2019 ). Case-control studies have shown the involvement of specific microorganisms in endogenous EtOH production, notably yeasts of the Candida and Pichia genus ( Mbaye et al., 2022 ) and bacteria including Klebsiella pneumoniae , Limosilactobacillus fermentum and Lactococcus lactis ( Mbaye et al., 2023 ; Yuan et al., 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Increased fecal ethanol and enriched ethanol-producing gut bacteria Limosilactobacillus fermentum, Enterocloster bolteae, Mediterraneibacter gnavus and Streptococcus mutans in nonalcoholic steatohepatitis

Mbaye,

Magdy Wasfy,

Borentain

et al. 2023

Front. Cell. Infect. Microbiol.

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BackgroundNon-alcoholic steatohepatitis (NASH) has become a major public health issue as one of the leading causes of liver disease and transplantation worldwide. The instrumental role of the gut microbiota is emerging but still under investigation. Endogenous ethanol (EtOH) production by gut bacteria and yeasts is an emerging putative mechanism. Microbial metagenomics and culture studies targeting enterobacteria or yeasts have been reported, but no culturomics studies have been conducted so far.AimTo assess fecal EtOH and other biochemical parameters, characterize NASH-associated dysbiosis and identify EtOH-producing gut microbes associated with the disease, fecal samples from 41 NASH patients and 24 controls were analyzed. High-performance liquid chromatography (HPLC) was used for EtOH, glucose, total proteins, triglyceride and total cholesterol. Viable bacteria were assessed with microbial culturomics. Microbial genetic material was assessed using 16S metagenomics targeting the hypervariable V3V4 region.ResultsFecal EtOH and glucose was elevated in the stools of NASH patients (p < 0.05) but not triglyceride, total cholesterol or proteins. In culturomics, EtOH-producing Enterocloster bolteae and Limosilactobacillus fermentum were enriched in NASH. V3V4 16S rRNA amplicon sequencing confirmed the enrichment in EtOH-producing bacteria including L. fermentum, Mediterraneibacter gnavus and Streptococcus mutans, species previously associated with NASH and other dysbiosis-associated diseases. Strikingly, E. bolteae was identified only by culturomics. The well-known Lacticaseibacillus casei was identified in controls but never isolated in patients with NASH (p < 0.05).ConclusionElevated fecal EtOH and glucose is a feature of NASH. Several different EtOH-producing gut bacteria may play an instrumental role in the disease. Culturomics and metagenomics, two complementary methods, will be critical to identify EtOH-producing bacteria for future diagnostic markers and therapeutic targets for NASH. Suppression of EtOH-producing gut microbes and L. casei administration are options to be tested in NASH treatment.

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The Characteristics of Gut Microbiota and Its Relation with Diet in Postmenopausal Osteoporosis

Chen,

Meng,

Wang

et al. 2024

Calcif Tissue Int

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Limosilactobacillus fermentum, Lactococcus lactis and Thomasclavelia ramosa are enriched and Methanobrevibacter smithii is depleted in patients with non-alcoholic steatohepatitis

Cited by 10 publications

References 57 publications

The Ethanol-Producing Enterocloster bolteae and Limosilactobacillus fermentum are enriched in Nonalcoholic steatohepatitis

The Ethanol-Producing Enterocloster bolteae and Limosilactobacillus fermentum are enriched in Nonalcoholic steatohepatitis

Increased fecal ethanol and enriched ethanol-producing gut bacteria Limosilactobacillus fermentum, Enterocloster bolteae, Mediterraneibacter gnavus and Streptococcus mutans in nonalcoholic steatohepatitis

The Characteristics of Gut Microbiota and Its Relation with Diet in Postmenopausal Osteoporosis

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