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Grant support

The authors thank grant AGL2017-89055-R, funded by MICIN/AEI and EU/FEDER; grant PID2023-147862OB-100 funded by MICIU/AEI and ERDF/EU; grant JDC2022-049517-I and grant RyC2021-034786-I, both funded by MICIU/AEI and EU/PRTR; and grant 20237AT026, funded by CSIC. The authors also thank Jesus Prados, Head of ICT Unit at CIAL, for his support in IT-related matters.

Analysis of institutional authors

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May 5, 2025
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Application of nanopore long-read sequencing and metabolomics in an in vitro dynamic intestinal digestion model: A genome-centric metatranscriptomic approach to investigating microbial TMA and SCFA metabolism

Publicated to:JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 262 (): 116896- - 2025-09-01 262(), DOI: 10.1016/j.jpba.2025.116896

Authors: Simo, Carolina; Mamani-Huanca, Maricruz; Hernandez-Hernandez, Oswaldo; Redondo-Rio, Alvaro; Munoz, Sergio; Garcia-Canas, Virginia

Affiliations

CNS, Barcelona Supercomp Ctr BSC, Barcelona, Spain - Author
Inst Res Biomed IRB, Barcelona, Spain - Author
UAM, Spanish Natl Res Council CSIC, Inst Food Sci Res CIAL, Mol Nutr & Metab, Madrid, Spain - Author

Abstract

The gut microbiota plays a relevant role in human health by metabolizing dietary components into bioactive molecules, including short-chain fatty acids and trimethylamine. Understanding how dietary interventions modulate microbial metabolism is key to developing strategies for reducing harmful metabolites such as TMA, a precursor of the pro-atherogenic trimethylamine-N-oxide. In this study, we integrated a dynamic in vitro gastrointestinal model (simgi (R)) with nanopore sequencing technology and metabolomics to investigate the impact of red thyme extract on microbial trimethylamine metabolism from L-carnitine. Metabarcoding, metagenomic, and metatranscriptomic analyses were performed alongside targeted metabolite quantification. Our results showed that microbial trimethylamine production primarily occurred in the transverse and descending colon compartments, coinciding with increased transcriptional activity of taxa harboring gbu cluster, associated with trimethylamine production. The administration of red thyme extract transiently reduced L-carnitine utilization but had a limited effect on overall trimethylamine levels. In parallel, short-chain fatty acids analysis revealed a shift in microbial fermentation patterns, with Acidaminococcus emerging as a dominant butyrate producer. Carbohydrate-active enzyme profiling identified Bacteroides and Parabacteroides genera as key mucin utilizers under the simulation conditions. These findings highlight the metabolic plasticity of the gut microbiota in response to the presence of L-carnitine and reduced complex carbohydrates availability, and provide new insights into microbial functional responses to dietary interventions targeting trimethylamine metabolism. Additionally, this study represents the first integration of nanopore-based metagenomics and genome-centric metatranscriptomics with targeted metabolomics in a dynamic in vitro gastrointestinal model. This multiomics approach enabled a detailed reconstruction of the microbial metabolic network involved in L-carnitine utilization and trimethylamine formation, offering a powerful tool for mechanistic studies of gut microbiota-diet interactions.

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Quality index

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-07-16:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 5.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 8 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 3.2.
  • The number of mentions on the social network X (formerly Twitter): 3 (Altmetric).

Leadership analysis of institutional authors

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: First Author (Simo, Carolina) and Last Author (García-Cañas, Virginia).

the author responsible for correspondence tasks has been García-Cañas, Virginia.