Sewage sludge is frequently used as a soil fertilizer although it may contain elevated concentrations of priority pollutants including di-(2-ethylhexyl)phthalate (DEHP). In the present study, the kinetics of microbial [ 14 C]DEHP mineralization was studied in laboratory microcosms with sewage sludge and agricultural soil. A biphasic model with two independent kinetic expressions was used to fit the mineralization data. The initial mineralization activity was described well by first-order kinetics (r 2 > 0.97), whereas mineralization in long-term incubations (>40 days) was described better by fractional power kinetics (r 2 > 0.95). The mineralization activity was much lower in the late phase presumably due to a decline in the bioavailability of DEHP caused by diffusion-limited desorption. The initial DEHP mineralization rate in sludge-amended soil varied between 3.7 and 20.3 ng of DEHP (g dw) -1 d -1 depending on incubation conditions. Aerobic DEHP mineralization was 4-5 times faster than anaerobic mineralization. DEHP mineralization in sludge-amended soil was much more temperature sensitive than was DEHP mineralization in soil without sludge. Indigenous microorganisms in the sewage sludge appeared to dominate DEHP degradation in sludgeamended soil. It was estimated that >41% of the DEHP in sludge-amended soil will have escaped mineralization after 1 year. In the absence of oxygen, >68% of the DEHP will not be mineralized within 1 year. Collectively, the data suggest that a significant fraction of the DEHP in sludgeamended soils may escape mineralization under in situ conditions.
The metabolism of phthalic acid (PA) and di-(2-ethylhexyl)phthalate (DEHP) in sludge-amended agricultural soil was studied with radiotracer techniques. The initial rates of metabolism of PA and DEHP (4.1 nmol/g [dry weight]) were estimated to be 731.8 and 25.6 pmol/g (dry weight) per day, respectively. Indigenous microorganisms assimilated 28 and 17% of the carbon in [14C]PA and [14C]DEHP, respectively, into microbial biomass. The rates of DEHP metabolism were much greater in sludge assays without soil than in assays with sludge-amended soil. Mineralization of [14C]DEHP to 14CO2 increased fourfold after inoculation of sludge and soil samples with DEHP-degrading strain SDE 2. The elevated mineralization potential was maintained for more than 27 days. Experiments performed with strain SDE 2 suggested that the bioavailability and mineralization of DEHP decreased substantially in the presence of soil and sludge components. The microorganisms metabolizing PA and DEHP in sludge and sludge-amended soil were characterized by substrate-specific radiolabelling, followed by analysis of 14C-labelled phospholipid ester-linked fatty acids (14C-PLFAs). This assay provided a radioactive fingerprint of the organisms actively metabolizing [14C]PA and [14C]DEHP. The14C-PLFA fingerprints showed that organisms with different PLFA compositions metabolized PA and DEHP in sludge-amended soil. In contrast, microorganisms with comparable 14C-PLFA fingerprints were found to dominate DEHP metabolism in sludge and sludge-amended soil. Our results suggested that indigenous sludge microorganisms dominated DEHP degradation in sludge-amended soil. Mineralization of DEHP and PA followed complex kinetics that could not be described by simple first-order equations. The initial mineralization activity was described by an exponential function; this was followed by a second phase that was described best by a fractional power function. In the initial phase, the half times for PA and DEHP in sludge-amended soil were 2 and 58 days, respectively. In the late phase of incubation, the apparent half times for PA and DEHP increased to 15 and 147 days, respectively. In the second phase (after more than 28 days), the half time for DEHP was 2.9 times longer in sludge-amended soil assays than in sludge assays without soil. Experiments with radiolabelled DEHP degraders suggested that a significant fraction of the 14CO2 produced in long-term degradation assays may have originated from turnover of labelled microbial biomass rather than mineralization of [14C]PA or [14C]DEHP. It was estimated that a significant amount of DEHP with poor biodegradability and extractability remains in sludge-amended soil for extended periods of time despite the presence of microorganisms capable of degrading the compound (e.g., more than 40% of the DEHP added is not mineralized after 1 year).
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