BackgroundGut microbiota alteration is implicated in the pathogenesis of alcoholic liver disease (ALD) and HCC. No study has characterized the dysbiosis associated with ALD by microbial culturomics, an approach that certifies viability and allows the characterization of pathobiont strain candidates.MethodsA case-control study was conducted on patients with ALD without HCC (ALD-NoHCC) (n=16), ALD with HCC (ALD-HCC) (n=19), and controls (n=24). 16S rRNA amplicon sequencing and microbial culturomics were used as complementary methods for gut microbiome profiling.ResultsBy microbial culturomics,Thomasclavelia ramosawas the most enriched and detected in all ALD samples (100%), while it was cultivated in only a small proportion of controls (20%, p < 0.001). By 16S rRNA amplicon sequencing and 3-groups linear discriminant analysis,T. ramosawas increased explicitly in the ALD-HCC group (LDA-score > 5, p < 0.05).ConclusionsT. ramosa,identified by culturomics and 16 rRNA sequencing, is associated with ALD and ALD-HCC. Alongside the recently reported in vitro genotoxicity of this species in colorectal cancer, this species has been identified as a candidate oncobiont in ALD-HCC.HighlightsThe gut microbiota signature of ALD and ALD-HCC was explored by microbial culturomics and 16S amplicon sequencingBy culturomics,T. ramosawas the most enriched and cultured from all included ALD patients, but in only 20% of controls (p < 0.05).T. ramosawas significantly associated with alcohol-related HCC by 16S sequencing.T. ramosais identified as a putative oncobiont associated with ALD-HCC, thus opening new avenues for diagnosis and treatment.Graphical abstract