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For investigation of main and interactive effects of six experimentally controlled environmental factors on phenol biodegradation in a shake-flask system, a largely neglected statistical procedure was applied. A major benefit resulting from the application of the orthogonal, fractional factorial design is that the number of experiments necessary to evaluate multifactor interactions is limited. In our investigation, the required number of experiments was reduced to 81 from the 324 necessary with conventional factorial designs; information was sacrificed for only 3 of 15 possible two-factor interactions. Six experimentally controlled factors were investigated at two or three treatment levels each; the six factors were (1) amount of phenol substrate, (2) amount of bacterial inoculum, (3) filtration of inoculum, (4) type of basal salts medium, (5) initial pH of basal salts medium, and (6) flask closure. Significant main effects were found for factors 1, 2, and 4; whereas significant interactive effects were found only for factor 2 with factor 3 and for factor 2 with factor 5. Our results suggest that the application of these statistical designs will greatly reduce the number of experiments necessary to evaluate multifactor effects on degradation rates during optimization of both hazard screening systems and waste treatment systems.

Use of a Fractional Factorial Design to Evaluate Interactions of Environmental Factors Affecting Biodegradation Rates