Development of Steady-State Diffusion Gradients for the Cultivation of Degradative Microbial Consortia

ABSTRACT

A diffusion gradient plate was constructed and evaluated for its potential use in the isolation of degradative microbial consortia from natural habitats. In this model, a steady-state concentration gradient of diclofop methyl, established by diffusion through an agarose gel, provided the carbon for microbial growth. Colonization of the gel surface was observed with epifluorescence and scanning confocal laser microscopy to determine microbial responses to the diclofop gradient. A detectable gradient developed over a narrow band (<10 mm). Consequently, quantitative analyses of the microbial response to the gradient were difficult to obtain. A two-dimensional, finite-element numerical transport model for advective-diffusive transport was used to simulate concentration and flux profiles in the physical model. The simulated profiles were correlated with the measured concentration gradient (R² = 0.89) and the cell numbers on the gel surface (R² = 0.85). The numerical model was subsequently used to redesign the physical model. The detectable concentration gradient in the modified physical model extended over the length of the gel (38 mm). The simulated profile again showed a good correlation with the measured profile (R² = 0.96) and the microbial responses to the concentration gradient (R² = 0.99). It was concluded that these gradients provide the steady-state environments needed to sustain steady-state consortia. They also provide a physical pathway for the development of degradative biofilms from low to high concentrations of toxicants and simulate conditions under which low concentrations of toxicant are supplied at a constant flux over long periods of time, such as the conditions that could occur in natural environments.