title: Constructing multispecies biofilms with defined compositions by sequential deposition of bacteria.
abstract: Rationally-assembled multispecies biofilms could benefit applied processes including mixed waste biodegradation and drug biosynthesis by combining complementary metabolic pathways into single functional communities. We hypothesized that the cellular composition of mature multispecies biofilms could be manipulated by controlling the number of each cell type present on newly colonized surfaces. To test this idea, we developed a method for attaching specific numbers of bacteria to a flow cell by recirculating cell suspensions. Initial work revealed a nonlinear relationship between suspension cell density and areal density when two strains of Escherichia coli were simultaneously recirculated; in contrast, sequential recirculation resulted in a predictable deposition of cell numbers. Quantitative analysis of cell distributions in 48-h biofilms comprised of the E. coli strains demonstrated a strong relationship between their distribution at the substratum and their presence in mature biofilms. Sequentially depositing E. coli with either Pseudomonas aeruginosa or Bacillus subtilis determined small but reproducible differences in the areal density of the second microorganism recirculated relative to its areal density when recirculated alone. Overall, the presented method offers a simple and reproducible way to construct multispecies biofilms with defined compositions for biocatalytic processes.
authors.affiliation: Department of Biology, Georgia State University, Atlanta, GA 30303, USA.
authors.forename: Bryan A
authors.initials: BA
authors.lastname: Stubblefield
authors.forename: Kristen E
authors.initials: KE
authors.lastname: Howery
authors.forename: Bianca N
authors.initials: BN
authors.lastname: Islam
authors.forename: Ariel J
authors.initials: AJ
authors.lastname: Santiago
authors.forename: Wendy E
authors.initials: WE
authors.lastname: Cardenas
authors.forename: Eric S
authors.initials: ES
authors.lastname: Gilbert
mesh_terms: D001412:Bacillus subtilis / Q000254:growth & development / Q000502:physiology*; D001422:Bacterial Adhesion; D001431:Bacteriological Techniques; D018441:Biofilms / Q000254:growth & development*; D015169:Colony Count, Microbial; D004783:Environmental Microbiology; D004926:Escherichia coli / Q000254:growth & development / Q000502:physiology*; D056265:Microbial Interactions; D011550:Pseudomonas aeruginosa / Q000254:growth & development / Q000502:physiology*