The simple two-chamber diffusion method was improved to study the diffusion properties of bacteriophage (phage) T4 through a model biofilm agarose gel membrane (AGM) embedded with dead host Escherichia coli K12 cells. The apparent diffusion coefficient (Dapp) of phage T4 was calculated to be 2.4 × 10−12 m2/s in 0.5% AGM, which was lower than the coefficient of 4.2 × 10−12 m2/s in 0.5% AGM without host cells. The phage adsorption process by dead host cells slowed the apparent phage diffusion. The Langmuir adsorption equation was used to simulate phage adsorption under different multiplicity of infections (MOIs); the maximum adsorbed phage MOI was calculated to be 417 PFU/CFU, and the Langmuir adsorption constant KL was 6.9 × 10−4 CFU/PFU. To evaluate the effects of phage proliferation on diffusion, a simple syringe-based biofilm model was developed. The phage was added into this homogenous biofilm model when the host cells were in an exponential growth phase, and the apparent diffusion coefficient was greatly enhanced. We concluded that Dapp of phages through biofilms could be distinctly affected by phage adsorption and proliferation, and that the idea of Dapp and these methods can be used to study diffusion properties through real biofilms. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012
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