The gastrointestinal tract (GIT) is responsible for essential digestive functions, which rely on
tight regulation by a network of neurons and glial cells embedded in the gut wall, termed the
enteric nervous system (ENS). These cells direct numerous gastrointestinal motor, immune
and homeostatic functions, which are vital for normal gut activity. A critical ENS-mediated
function is the coordination of gastrointestinal smooth muscle to generate force for gut
transit. Consequently, ENS dysregulation and functional dysmotility underlie various
gastrointestinal disorders. However, whether ENS dysregulation and altered gut motility
contribute to enteric infection has not been as greatly explored. Therefore, we aimed to
determine if infection with Clostridioides difficile, a pathogen that causes extensive colonic
damage, affects the ENS and explore how ENS dysregulation contributes to disease. Using
a mouse model of C. difficile infection (CDI), we demonstrated that disease perturbed
smooth muscle and ENS architecture. To link this structural disorder with functional motor
alteration, we showed that CDI decreased the contractive ability of the colon, resulting in a
slowing of fecal transit through the GIT. These results demonstrate that CDI causes
considerable functional dysmotility by inducing smooth muscle and ENS structural
derangement. Importantly, alterations to gastrointestinal transit may have subsequent
consequences on gut microbiota activity, disease symptoms, and recurrence risk. Therefore,
this work could aid in the generation of new treatments targeting the smooth muscle and
ENS to restore gut function, and combat disease phenotypes resulting from gastrointestinal
infections.