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Applied and Environmental Microbiology, December 1998, p. 4711-4719, Vol. 64, No. 12
Environmental Engineering Laboratory, Aalborg
University, DK-9000 Aalborg, Denmark
Received 14 July 1998/Accepted 17 September 1998
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. The
14C-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).
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Degradation of Phthalate and Di-(2-Ethylhexyl)phthalate by
Indigenous and Inoculated Microorganisms in Sludge-Amended
Soil
*
Corresponding author. Mailing address: Environmental
Engineering Laboratory, Aalborg University, Sohngaardsholmsvej 57, DK-9000 Aalborg, Denmark. Phone: 45 96 358505. Fax: 45 98142555. E-mail: pr{at}civil.auc.dk.
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