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A mixed culture with the ability to aerobically biodegrade 1,4-dioxane in the presence of tetrahydrofuran (THF) was enriched from a 1,4-dioxane contaminated aquifer. This consortium contained 3–4 morphologically different types of colonies and was grown in mineral salts media. Biodegradation of 1,4- dioxane began when THF concentrations in batch experiments became relatively low. No biodegradation of 1,4-dioxane was observed in the absence of THF and the measured cell yield was similar during degradation of 1,4-dioxane with THF or with THF alone. However, when the consortium was grown in the presence of 14C-1,4-dioxane plus THF, 2.1% of the radiolabeled 1,4-dioxane was present in the particulate fraction. The majority of the 14C (78.1%) was recovered as 14CO2, while 5.8% remained in the liquid fraction. This activity is interesting since the non-growth substrate is mineralized, yet only minimally assimilated into biomass. Using THF as the growth substrate, 1,3-dioxane, methyl t-butyl ether, ethyl t-butyl ether and t-amyl methyl ether.