Engineering a synthetic gut model to explore Clostridioidesdifficile infections

Our intestines play home to over one thousand bacterial species, often referred to as the gut microbiota. This microbiota profoundly affects our bodies, providing an array of benefits. Disturbances to this community are associated with pathogenic infection and diseases such as obesity, diabetes and inflammatory bowel disease. Our aim is to engineer a synthetic microbiota, with an emphasis on tracking individual species. Introducing this community into our in vitro gut epithelial culture system will provide insights into pathogen infection strategies, and mechanisms by which the microbiota generates resistance. This tool can be used for the screening of drug/probiotic candidates, potentially reducing the need for animals. Using PMA-qPCR we are able to track up to ten single species in a mixed biofilm. Clostridium difficile an opportunistic pathogen targets the gut during times of depleted microbiota. We have begun using our tracking technology to see how a representative microbiota reacts to a C. difficile invasion. Alongside this, we investigated the effect of the gut commensal - Bacteroides dorei on C. difficile utilizing classical quantification. In both a standard biofilm and in our in vitro model, we see a reduced number of C. difficile when in mixed culture.