Characterization of Biofilm Formation by Environmental Clostridioides difficile Isolates

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Clostridioides difficile is a gram-positive, spore-forming bacteria capable of causing disease, referred to as Clostridioides difficile Infections (CDI), which may include symptoms such as severe diarrhea and colitis. Every year in the United States, about half a million of these CDI cases result in approximately $6 billion in medical costs. C. difficile is difficult to eradicate due to its antibiotic resistance, virulent properties, and the fact that there is no available vaccine. CDIs are thought to be iatrogenic, but accumulating evidence suggests that environmental transmission may play an important role as well. Previously, we were able to isolate multiple C. difficile samples from fish markets, hospital wastewater, and wastewater treatment plants in southern Taiwan. Genotypic analysis revealed the presence of toxigenic C. difficile isolates closely related to ones prevalent among humans and animals infected by C. difficile. To gain a better understanding of the virulence capabilities of these environmental isolates, a series of studies including growth rates, spore production, and cytotoxicity were performed; however, the biofilm forming abilities were not well characterized. In this study, biofilm formation ability of these isolates was observed over a 72-hour period, then biofilm mass was measured using a crystal violet staining assay. By using a One-Way ANOVA, results were compared to the control lab strains 630 and R20291 to identify any significant differences. Further comparisons were made between isolates based on biofilm morphology. We observed a diverse range in biofilm formation abilities as well as biofilm morphology, but we did not detect any significant correlation between the robustness of the biofilms and the presence of toxin genes. Ongoing experiments focus on assessing the ability of these biofilms to resist antibiotics. In summary, our preliminary results contribute to the characterization of multiple environmental C. difficile isolates obtained from water and seafood samples in southern Taiwan.
Original languageAmerican English
StatePublished - 4 Nov 2022
Event111th Annual Technical Meeting, Oklahoma Academy of Science - Oklahoma State University Center for Health Sciences, Tulsa, United States
Duration: 4 Nov 20224 Nov 2022


Conference111th Annual Technical Meeting, Oklahoma Academy of Science
Abbreviated titleOAS Meeting 2022
Country/TerritoryUnited States
Internet address


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