This study investigates the metabolic interactions between Clostridioides difficile and Enterococcus faecalis in the cecum of mice. Using MALDI imaging mass spectrometry, the research highlights how E. faecalis influences C. difficile pathogenesis through arginine metabolism.
Take tissue cryosections from the cecum, a region of the large intestine. One section is from a mouse infected only with Clostridioides difficile. The other section is from a mouse co-infected with C. difficile and Enterococcus faecalis.
In the co-infected tissue, E. faecalis metabolizes host arginine and releases ornithine. C. difficile absorbs this ornithine, promoting growth-associated toxin production and tissue damage.
To analyze these samples using MALDI imaging, apply a uniform layer of matrix solution.
Load the slides into a MALDI mass spectrometer, where a laser scans the tissue, ionizing amino acids through the matrix.
Load the slides into a MALDI mass spectrometer, where a laser scans the tissue, ionizing amino acids through the matrix.
The resulting ions are separated by their mass-to-charge ratio and detected by the instrument to create spatial maps of amino acid distribution.
The co-infected tissue shows lower arginine and higher ornithine levels than the monoinfected sample, which supports microbial cooperation–driven C. difficile pathogenesis.
Perform the MALDI matrix application and MALDI imaging mass spectrometry as demonstrated earlier.
Analyze the ion images from multiple biological replicates, and compare the results for a significance via intensity box plot comparisons using the SCiLS statistical software.
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