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Histological study of diurnal changes in bacterial settlement in the rat alimentary tract

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Abstract

The composition of fecal bacteria is reported to change throughout the day, whereas the circadian rhythmicity of indigenous bacteria that settle on the epithelium is mostly unknown. The present study aimed to clarify the diurnal changes in the settlement of indigenous bacteria in the rat alimentary tract using histological analysis. The settlement of indigenous bacteria on the mucosal epithelium throughout the day and the diurnal changes in settlement levels were observed in the esophagus, the nonglandular area of the stomach, and the ileum. The peak of zeitgeber time (ZT) in the settlement level differed by segment: ZT 12 in the esophagus, ZT 6 in the nonglandular area of the stomach, and ZT 0 in the ileum. Moreover, 16S rRNA amplicon sequencing using tissue sections revealed that the compositions of the indigenous bacteria in the ileum differed among ZT. In the intervillous spaces of the ileum, the formation level of the mucus layer, one of the most fundamental host defenses against bacteria, was lowest at ZT 0. Bacteria were preferentially adjacent to the villous epithelium in the area without coverage by the mucus layer at ZT 0. These findings collectively suggest that the settlement level and possibly the composition of the indigenous bacteria changed diurnally in various segments of the alimentary tract, and the formation of the mucus layer might be the most likely to lead to such diurnal changes in indigenous bacteria, at least in the ileum.

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Acknowledgements

Gram-positive Staphylococcus epidermidis and Gram-negative Escherichia coli for evaluating the quantity of Gram-positive bacteria were kindly provided by Dr. Eiko Matsuo (Kobe University).

Funding

This study was supported by the Japan Society for the Promotion of Science (grant numbers: 16K18813 and 20K15902) and by the Foundation of Kinoshita Memorial Enterprise.

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Authors and Affiliations

  1. Laboratory of Histophysiology, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo, 657-8501, Japan

    Nanami Sakata, Youhei Mantani, Satoki Nakanishi & Rinako Morishita

  2. Laboratory of Animal Molecular Morphology, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo, 657-8501, Japan

    Toshifumi Yokoyama & Nobuhiko Hoshi

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  1. Nanami Sakata
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  2. Youhei Mantani
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  5. Toshifumi Yokoyama
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  6. Nobuhiko Hoshi
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Correspondence to Youhei Mantani.

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This study was approved by the Institutional Animal Care and Use Committee (permission number 30–05-01). All procedures in studies involving animals were performed in accordance with the ethical standards of the institution (the Kobe University Animal Experimentation Regulations) or at the practice at which the studies were conducted.

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Sakata, N., Mantani, Y., Nakanishi, S. et al. Histological study of diurnal changes in bacterial settlement in the rat alimentary tract. Cell Tissue Res 389, 71–83 (2022). https://doi.org/10.1007/s00441-022-03626-9

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