Abstract

Recent evidence suggests that gut microbiota shifts can alter host metabolism even during healthy aging. Lactobacillus acidophilus DDS-1, a probiotic strain, has shown promising probiotic character in vitro, as well as in clinical studies. The present study was carried out to investigate whether DDS-1 can modulate the host metabolic phenotype under the condition of age-affected gut microbial shifts in young and aging C57BL/6J mice. Collected fecal samples were analyzed using 16S rRNA gene sequencing for identifying gut microbiota and untargeted gas chromatography-mass spectrometry (GC-MS) metabolomics analysis. Gut microbial shifts were observed in the control groups (young and aging), leading to an alteration in metabolism. Principal coordinate analysis (PCoA) of microbiota indicated distinct separation in both the DDS-1-treated groups. L. acidophilus DDS-1 increased the relative abundances of beneficial bacteria, such as Akkermansia muciniphila and Lactobacillus spp., and reduced the relative levels of opportunistic bacteria such as Proteobacteria spp. Metabolic pathway analysis identified 10 key pathways involving amino acid metabolism, protein synthesis and metabolism, carbohydrate metabolism, and butanoate metabolism. These findings suggest that modulation of gut microbiota by DDS-1 results in improvement of metabolic phenotype in the aging mice.

Highlights

  • Microbes residing in a mammalian gut co-evolve with age

  • Of all the pathways identified, two pathways were found to be common among both age groups, galactose metabolism and alanine, aspartate, and glutamate metabolism

  • We used a combination of 16S rRNA sequencing analysis and untargeted metabolomics profiling of fecal samples to identify specific gut microbiota changes and numerous gut metabolites that were associated with response to DDS-1 supplementation in young and aging mice

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Summary

Introduction

Microbes residing in a mammalian gut co-evolve with age. These microbes, known as the ‘gut microbiota’, interact with each other, as well as the host, and impact the health of the host [1]. The abundance of beneficial bacteria, such as Akkermansia spp. and Lactobacillus spp., were reduced in the aging population [7,8,9] Such drastic microbial shifts in healthy-aging could lead to increased intestinal inflammation and changes in host metabolism. Along these lines, a few animal and clinical studies demonstrated metabolic trajectory associated with gut microbiome upon aging [10,11,12,13]. The above-mentioned studies [18,24,27] encouraged us to hypothesize that the probiotic strain, L. acidophilus DDS-1, could further improve the beneficial microbial composition of healthy gut and host metabolism. A combination of omics-based approaches, involving 16S rRNA gene sequencing and untargeted GC-MS based metabolomics, were utilized to provide a comprehensive understanding of changes in host metabolic phenotype of altering gut microbiota with and without DDS-1 supplementation

Ethics Statement
Bacterial Culture and Probiotic Feed Preparation
Animals and Probiotic Treatments
Fecal Sample Collection and Preparation
Microbiota Analysis Using 16S rRNA High-Throughput Sequencing
Fecal Metabolomics
Data Analysis and Multivariate Analysis
Statistical Analysis
Gut Microbial Changes at the Phylum Level
Kruskal-Wallis
Gut Microbial Changes at Genus and Species Level Among Four Groups
Metabolic Phenotyping of YC and YP Groups
Metabolic
Comparative Metabolite Phenotyping in Young and Aging Groups
Discussion
Amino Acid and Protein Metabolism
Carbohydrate Metabolism
Butanoate Metabolism
Conclusions
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