The type 3 fimbriae of Klebsiella pneumoniae are comprised of the major fimbrial subunit (MrkA) and the adhesin (MrkD) that has previously been shown to mediate binding to collagen. The ability of adhesive and non-adhesive derivatives of K. pneumoniae to form biofilms on collagen-coated surfaces in continuous-flow chambers was investigated. Unlike biofilm formation on abiotic plastic surfaces, the presence of the MrkD adhesin was necessary for growth on collagen-coated surfaces. Fimbriate strains lacking the MrkD adhesin did not efficiently adhere to and grow on these surfaces. Similarly, purified human extracellular matrix and the extracellular matrix formed by human bronchial epithelial cells grown in vitro provided a suitable substrate for MrkD-mediated biofilm formation, whereas direct binding to the respiratory cells was not observed. Type 3 fimbriae may therefore have two roles in the early stages of adherence and growth on in-dwelling devices such as endotracheal tubes. The MrkA polypeptide could facilitate adsorption to abiotic polymers of recently implanted devices and the MrkD adhesin could enable bacteria to adhere to and grow on polymers coated with host-derived proteins.