Use of high throughput amplicon sequencing and ethidium monoazide dye to track microbiota changes in an equol-producing menopausal woman receiving a long-term isoflavones treatment

Lucía Guadamuro; M. Andrea Azcárate-Peril; Rafael Tojo; Baltasar Mayo; Susana Delgado; Lucía Guadamuro; M. Andrea Azcárate-Peril; Rafael Tojo; Baltasar Mayo; Susana Delgado

doi:10.3934/microbiol.2019.1.102

AIMS Microbiology

2019, Volume 5, Issue 1: 102-116. doi: 10.3934/microbiol.2019.1.102

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Use of high throughput amplicon sequencing and ethidium monoazide dye to track microbiota changes in an equol-producing menopausal woman receiving a long-term isoflavones treatment

1.
Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Paseo Río Linares s/n, 33300-Villaviciosa, Asturias, Spain
2.
Department of Medicine, Division of Gastroenterology and Hepatology, and Microbiome Core Facility, School of Medicine, University of North Carolina (UNC), Chapel Hill, NC 27599-7555, USA
3.
Gastroenterology Department, Cabueñes University Hospital, 33203-Gijón, Asturias, Spain

Received: 17 October 2018 Accepted: 26 February 2019 Published: 22 March 2019

This work describes the impact of long term consumption of an isoflavone-rich dietary daily supplement on the composition and diversity of the faecal microbiota of a menopausal, equol-producing woman. Sequencing of 16S rDNA amplicons was performed on faecal samples taken at 0, 1, 3 and 6 months of treatment. Additionally, and for comparative purposes, ethidium monoazide (EMA) was used to avoid detection of DNA from dead bacteria. Members of two genera of the family Coriobacteriaceae (Eggerthella and Collinsella) were found in greater proportions at all sampling points during isoflavone supplementation. Different genera of the family Ruminococcaceae (e.g., Ruminococcus and Faecalibacterium), as well as members of the family Lachnospiraceae (Coprococcus) also underwent significant increases. For this last genus a positive correlation with the levels of equol excretion in urine was found. Currently bacterial strains known to be involved in isoflavone metabolism and equol production have been assigned to these taxa. The use of EMA dye allowed us to unravel those bacterial gut linages (e.g., Lachnospiraceae) that were more susceptible to damage. Our study contributes to the identification of microorganisms possibly involved in the production of isoflavone-desirable metabolites (such as equol), which could ultimately be isolated and further used as probiotics by people who cannot naturally benefit from isoflavones.
- gut microbiota,
- isoflavones,
- equol,
- pyrosequencing,
- ethidium monoazide,
- menopause,
- metabotype
Citation: Lucía Guadamuro, M. Andrea Azcárate-Peril, Rafael Tojo, Baltasar Mayo, Susana Delgado. Use of high throughput amplicon sequencing and ethidium monoazide dye to track microbiota changes in an equol-producing menopausal woman receiving a long-term isoflavones treatment[J]. AIMS Microbiology, 2019, 5(1): 102-116. doi: 10.3934/microbiol.2019.1.102

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Abstract

This work describes the impact of long term consumption of an isoflavone-rich dietary daily supplement on the composition and diversity of the faecal microbiota of a menopausal, equol-producing woman. Sequencing of 16S rDNA amplicons was performed on faecal samples taken at 0, 1, 3 and 6 months of treatment. Additionally, and for comparative purposes, ethidium monoazide (EMA) was used to avoid detection of DNA from dead bacteria. Members of two genera of the family Coriobacteriaceae (Eggerthella and Collinsella) were found in greater proportions at all sampling points during isoflavone supplementation. Different genera of the family Ruminococcaceae (e.g., Ruminococcus and Faecalibacterium), as well as members of the family Lachnospiraceae (Coprococcus) also underwent significant increases. For this last genus a positive correlation with the levels of equol excretion in urine was found. Currently bacterial strains known to be involved in isoflavone metabolism and equol production have been assigned to these taxa. The use of EMA dye allowed us to unravel those bacterial gut linages (e.g., Lachnospiraceae) that were more susceptible to damage. Our study contributes to the identification of microorganisms possibly involved in the production of isoflavone-desirable metabolites (such as equol), which could ultimately be isolated and further used as probiotics by people who cannot naturally benefit from isoflavones.

Acknowledgments

The study was partially supported by projects from the Spanish Ministry of Economy and Competitiveness (MINECO) (AGL-2014-57820-R) and Asturias Principality (GRUPIN14-137). The Microbiome Core Facility is supported in part by the NIH/National Institute of Diabetes and Digestive and Kidney Diseases grant P30 DK34987. LG was supported by a research contract of the FPI Program from MINECO (BES-2012-062502). SD was supported by a research contract from MINECO associated to the project BIO2014-55019-JIN.

Conflict of Interest

The authors declare that there is no conflict of interests regarding the publication of this article.

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