Constructing multispecies biofilms with defined compositions by sequential deposition of bacteria

• Published in: Applied microbiology and biotechnology 2010-05, Vol.86 (6), p.1941-1946
• Main Authors: Stubblefield, Bryan A, Howery, Kristen E, Islam, Bianca N, Santiago, Ariel J, Cardenas, Wendy E, Gilbert, Eric S
• Format: Article
• Language: English
• Subjects: Applied Microbial and Cell Physiology; Bacillus subtilis; Bacillus subtilis - growth & development; Bacillus subtilis - physiology; Bacteria; Bacterial Adhesion; Bacteriological Techniques; Biocatalysts; Bioconversions. Hemisynthesis; Biodegradation; Biofilms; Biofilms - growth & development; Biological and medical sciences; Biomedical and Life Sciences; Biosynthesis; Biotechnology; Catalysis; Colony Count, Microbial; E coli; Environmental Microbiology; Escherichia coli; Escherichia coli - growth & development; Escherichia coli - physiology; Experiments; Fundamental and applied biological sciences. Psychology; Life Sciences; Metabolism; Methods. Procedures. Technologies; Microbial Genetics and Genomics; Microbial Interactions; Microbiology; Microscopy; Pseudomonas aeruginosa; Pseudomonas aeruginosa - growth & development; Pseudomonas aeruginosa - physiology; Quantitative analysis; Studies
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-010-2473-y

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.