Using a ligate intestinal loop mouse model to investigate Clostridioides difficile adherence to the intestinal mucosa in aged mice.

Pablo Castro-Córdova,María José Mendoza-León,Daniel Paredes-Sabja,Yung-Fu Chang

doi:10.1371/journal.pone.0261081

Pablo Castro-Córdova, María José Mendoza-León + Show 2 more

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https://doi.org/10.1371/journal.pone.0261081

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Abstract

Interaction of Clostridioides difficile spores with the intestinal mucosa contributes to the persistence and recurrence of the infection. Advanced age is one of the main risk factors for C. difficile infection and recurrence of the disease. However, interaction of C. difficile spores with the intestinal mucosa during aging has not been evaluated. In the present work, using intestinal ligated loop technique in a mouse model, we analyzed C. difficile spore adherence and internalization to the ileum and colonic mucosa during aging. Additionally, we provide visual documentation of the critical steps of the procedure. Consequently, our data suggest that spore internalization in the ileum and colonic mucosa is higher in elderly mice rather than adults or young mice. Also, our data suggest that spore adherence to the ileum and colonic mucosa decreases with aging.

Highlights

Clostridioides difficile is a Gram-positive, anaerobic, and spore former bacterium and the leading pathogen causing hospital acquiring diarrhea associated with antibiotics [1,2]
We provide a detailed method with video, to quantify spore adherence and internalization in the ileum and colonic mucosa of mice, in particular, we detailed a surgical procedure of intestinal ligated loop technique, including animal anesthetize, opening of peritoneal cavities, the performance of ligated intestinal loop with inoculation of C. difficile spores, incision suturing and animal necropsy, followed by fixation of the tissues to whole-mounted tissue immunofluorescence and mounting of the sample for visualization by confocal microscopy
Using the intestinal ligated loop technique, here, we describe that adherence of C. difficile spores to the ileum, and colonic mucosa is decreased in mice of 1-years-old and 2-years-old compared to 7-weeks-old mice

Summary

Introduction

Clostridioides difficile is a Gram-positive, anaerobic, and spore former bacterium and the leading pathogen causing hospital acquiring diarrhea associated with antibiotics [1,2]. The two main risk factors for CDI are the continuous alteration of the intestinal microbiota caused by antibiotics, and the age over 65 years old [7,8], being the 91% of the CDI deaths in this age group [9]. This increasing association of CDI with aging could be explained by agerelated physiologic changes such as the immunosenescence and age-related dysbiosis of the intestinal microbiota; reducing protection against C. difficile

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