Skip to main content
ARS Home » Northeast Area » Wyndmoor, Pennsylvania » Eastern Regional Research Center » Dairy and Functional Foods Research » Research » Publications at this Location » Publication #414984
Research Project: In vitro Human Gut System: Interactions Between Diet, Food Processing, and Microbiota

Location: Dairy and Functional Foods Research

Title: Bacterial metabolites influence the autofluorescence of Clostridioides difficile

Author
item TICER, TAYLOR - Medical University Of South Carolina
item GLOVER, JANIECE - Medical University Of South Carolina
item DOOLEY, SARAH - Medical University Of South Carolina
item KENDRICK, JACOB - Medical University Of South Carolina
item ZACKULAR, JOSEPH - University Of Pennsylvania
item DEVKOTA, SUZANNE - Cedars-Sinai Medical Center
item Mahalak, Karley
item WU, GARY - University Of Pennsylvania
item ENGEVIK, AMY - Medical University Of South Carolina
item ENGEVIK, MELINDA - Medical University Of South Carolina
item TINGLER, ANNA - Medical University Of South Carolina

Submitted to: Frontiers in Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/2/2024
Publication Date: 10/8/2024
Citation: Ticer, T.D., Glover, J.S., Dooley, S.A., Kendrick, J., Zackular, J., Devkota, S., Mahalak, K.K., Wu, G., Engevik, A., Engevik, M., Tingler, A. 2024. Bacterial metabolites influence the autofluorescence of Clostridioides difficile. Frontiers in Microbiology. https://doi.org/OI 10.3389/fmicb.2024.1459795.
DOI: https://doi.org/10.3389/fmicb.2024.1459795

Interpretive Summary: Clostridioides difficile is a bacterium responsible for a range of gastrointestinal illnesses that is also able to auto-fluoresce under certain conditions. This property of C. difficile could be very useful for researchers wanting to identify it in mixed bacterial populations both for laboratory work and for identification of its presence in patient samples. To understand this property, we used a variety of bacterial growth and screening methods to identify key chemical compounds present in cultures that increased the level of fluorescence of C. difficile. We also found that certain bacteria, especially Klebsiella pneumoniae, can also produce these chemical compounds, increasing the autofluorescence of C. difficile when grown together. Using techniques to separate bacteria that fluoresce differently, we were able to separate C. difficile from K. pneumoniae using this autofluorescence property. This is a first step towards understanding how this fluorescence occurs on a technical level.

Technical Abstract: Clostridioides difficile is bacterial pathogen that has been implicated in severe gastrointestinal infections. C. difficile has intrinsic green autofluorescence and the level of this autofluorescence is known to be increased by growth time and oxygen. Currently, it is unclear if dietary compounds or metabolites from the gut microbiota are able to enhance C. difficile autofluorescence. Here, we aimed to determine potential factors that affect C. difficile autofluorescence. After screening a large repertoire of compounds, we identified several substances, like L-lysine and pantothenate, that led to an increased C. difficile autofluorescence. We also found that several members of the gut microbiota, such as Enterococcus faecalis, Klebsiella aerogenes and K. pneumoniae, can increase C. difficile autofluorescence through their secreted compounds. We further focused on the effect of K. pneumoniae on C. difficile autofluorescence and found that multiple enteric strains of K. pneumoniae could enhance C. difficile’s autofluorescence. We further used this enhanced autofluorescence to identify C. difficile in K. pneumoniae co-cultures by flow cytometry. Our findings shed light on the relationship between C. difficile and other members of the gut microbiota, as well as different factors that can affect C. difficile autofluorescence.