Spatial Distribution and Coexistence of Klebsiella pneumoniae and Pseudomonas aeruginosa in Biofilms

The heterotrophic bacteria Klebsiella pneumoniae and Pseudomonas aeruginosa stably coexisted in laboratory-grown biofilms, even though the growth rate of K. pneumoniae was twice that of P. aeruginosa under planktonic growth conditions. The failure of K. pneumoniae to displace P. aeruginosa from the biofilm could not be attributed to concentration gradients of the limiting nutrient (glucose) arising from the interaction of reaction and diffusion. Comparisons of the growth rates of the two species in mono- and binary-population biofilms suggested partial segregation of the two species in the latter. We used a fluorescently labeled monoclonal antibody to examine the spatial distribution of K. pneumoniae in frozen cross sections of biofilm to confirm this segregation. K. pneumoniae microcolonies resided on top of, or intermixed with, a base film of P. aeruginosa. We hypothesize that microscale structural heterogeneity and differing rates of bacterial attachment and detachment of the two species are responsible for coexistence in this system.