Alterations to the gut microbiome caused by changes in diet, consumption of antibiotics, etc., can affect host function. Moreover, perturbation of the microbiome during critical developmental periods potentially have long-lasting impacts on hosts. Using four selectively bred High Runner and four non-selected Control lines of mice, we examined the effects of early-life diet and exercise manipulations on the adult microbiome by sequencing the hypervariable Internal Transcribed Spacer region of the bacterial gut community. Mice from High Runner lines run ∼3-fold more on wheels than do Controls, and have several other phenotypic differences (e.g., higher food consumption and body temperature) that could alter the microbiome, either acutely or in terms of coevolution. Males from generation 76 were given wheels and/or Western diet from weaning until sexual maturity at 6 weeks of age, then housed individually without wheels on standard diet until 14 weeks of age, when fecal samples were taken. Juvenile Western diet reduced bacterial richness and diversity after the 8-week washout period (equivalent to ∼6 human years). We also found interactive effects of genetic linetype, juvenile diet, and/or juvenile exercise on microbiome composition and diversity. Microbial community structure clustered significantly in relation to both linetype and diet. Western diet also reduced the relative abundance of Muribaculum intestinale. These results constitute one of the first reports of juvenile diet having long-lasting effects on the adult microbiome after a substantial washout period. Moreover, we found interactive effects of diet with early-life exercise exposure, and a dependence of these effects on genetic background.

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Increases in availability of energy‐dense foods and simultaneous reductions in physical activity of Western industrialized societies has created an environment that promotes obesity, but alterations in the gut microbiome may also play a role. As with other organismal characteristics, the microbiome may be influenced by factors experienced early in life. We previously demonstrated that early‐life wheel access for 3 weeks, starting at weaning, followed by an 8‐weeks washout period, increases adult wheel running in both selectively bred High Runner (HR) and non‐selected Control (C) lines of mice. We extended these studies by examining effects of early‐life treatments on the gut microbiome. Mice were given early‐life wheel access and/or Western diet from weaning until sexual maturation at 6 weeks of age, then housed individually without wheels and on standard diet until 14 weeks of age, when fecal samples were taken. We profiled the gut microbiome by extracting DNA from fecal samples using MoBio Laboratories PowerSoil DNA Isolation kit, amplified ITS rRNA, sequenced the libraries using an Illumina MiSeq, and chose OTUs using Usearch. For bacterial richness (number of OTUs and Chao1 index) we found a 3‐way interaction of linetype, juvenile Western diet, and juvenile wheel access. These results constitute one of the first reports of juvenile exercise and/or diet having long‐lasting effects on the adult gut microbiome after a substantial washout period (equivalent to ~6 human years). Support or Funding Information Supported in part by NIH grant R21HD084856

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