The increasing trend of community-acquired infections associated with C. difficile, a Gram (+), obligate anaerobe, and spore-forming bacterium known as a nosocomial agent, indicates the importance of considering other contamination sources of this pathogen. The growing population, changing climate conditions, rising sea temperatures, mucilage formation, flood content mixing with drinking water resources and sewage discharge increase the likelihood of C. difficile contamination in aquatic ecosystems. Seafood in these contaminated water environments is naturally at risk of microbial contamination, posing a significant public health concern. Currently, efforts to combat C. difficile infections are largely focused on diagnosis and treatment. However, adopting an effective multidisciplinary approach under the One Health perspective will facilitate the implementation of sustainable and solution-oriented measures in infection management. This review comprehensively examines the presence of C. difficile in aquatic ecosystems and seafood, emphasizing the importance of environmental sources in controlling infections. Thus, by adopting the One Health approach, it aims to contribute to the development of effective and sustainable solutions for controlling C. difficile infections. Additionally, methods used for isolating C. difficile from seafood are reviewed, and recommendations for future researchers on isolating C. difficile from food are provided.