e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183

available at www.sciencedirect.com

journal homepage: www.elsevier.com/locate/ecoleng

A comparative study of surface and subsurface flow

constructed wetlands for treatment of combined sewer
overflows: A greenhouse experiment

Annelies M.K. Van de Moortel a,∗ , Diederik P.L. Rousseau b ,

Filip M.G. Tack a , Niels De Pauw c
a Department of Applied Analytical and Physical Chemistry, Laboratory of Analytical Chemistry and Applied Ecochemistry, Ghent
University, Coupure Links 653, 9000 Ghent, Belgium
b Department of Environmental Resources, UNESCO-IHE Institute for Water Education, P.O. Box 3015, 2601 DA Delft, The Netherlands
c Department of Applied Ecology and Environmental Biology, Laboratory of Environmental Toxicology and Aquatic Ecology, Ghent

University, J. Plateaustraat 22, 9000 Ghent, Belgium

a r t i c l e i n f o a b s t r a c t

Article history: The use of surface flow (SFCWs) and subsurface flow constructed wetlands (SFCWs) for the
Received 7 December 2007 treatment of combined sewer overflows was assessed at pilot scale. Synthetic wastewa-
Received in revised form ter was applied in three batches with decreasing concentrations to mimic concentration
27 August 2008 profiles that are obtained in the field during overflow events. Three simulated combined
Accepted 29 August 2008 sewer overflows were applied on each wetland. Composite water samples (60 in total) were
taken for a period of 8 days to study the removal of total nitrogen (Ntot), NH4 –N, NO3 –N,
total COD (CODtot) and total phosphorus. Redox potential, which was monitored at vari-
Keywords: ous locations along the wetlands, was more negative in the SSFCWs. In general, removal
Waste water treatment occurred faster in the SSFCWs and the final concentrations were lower. The removal of Ntot
Ammonium was only 36.6 ± 3.3% in the SFCWs due to nitrification-limiting conditions. The conditions in
Nitrogen the SSFCWs, in contrast, seemed to promote Ntot removal (removal efficiency 96.7 ± 1.9%).
Phosphorus The removal of P was hampered in both wetland types by reducing conditions. P that was
Phragmites initially removed was released again from the substrates later on. First-order removal rate
Fractionation constants were derived for the removal of both CODtot (SSFCWs: 1.1 ± 0.3 m d−1 ; SFCWs:
Treatment wetlands 0.17 ± 0.06 m d−1 ) and Ntot (SSFCWs: 0.4 ± 0.1 m d−1 ; SFCWs: 1.7 ± 0.5 m d−1 ).
Stormwater © 2008 Elsevier B.V. All rights reserved.
k–C* model
First-order removal

1. Introduction nearest surface water, thereby potentially causing a sudden

During rain events, the capacity of combined sewer systems
may be too confined to allow transport of all waste- and
stormwater to a treatment plant. Combined sewer overflows
(CSOs) are provided for diverting the excess water to the
pollution shock. The study of devices applicable for the tempo-
ral storage and treatment of stormwater and combined sewer
overflows has been pointed out as one of the 10 priority areas
in urban wet weather flow research (Heaney et al., 1999). The
occurrence of uncontrolled or poorly controlled discharges

∗
Corresponding author. Tel.: +32 9 264 59 95.
E-mail addresses: Annelies.Vandemoortel@UGent.be (A.M.K. Van de Moortel), d.rousseau@unesco-ihe.org (D.P.L. Rousseau).
0925-8574/$ – see front matter © 2008 Elsevier B.V. All rights reserved.
doi:10.1016/j.ecoleng.2008.08.015
176 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183

from CSOs and stormwater is one of the major reasons for

Table 1 – Composition of the applied synthetic waste
the long-term persistence of the poor water quality of surface water with concentration C1 (modified after Boeije et al.,
waters (Suarez and Puertas, 2005; Robson et al., 2005). 1998).
The use of constructed wetlands for treatment of domestic
Compound mg l−1
wastewater has been studied intensively (Kadlec and Knight,
1996; Scholz et al., 2002; Mungasavalli and Viraraghavan, 2006; MgCl2 51.0
Vymazal, 2007). More recently wetlands have been used for CaCl2 107.5
Urea 58.6
the retention and treatment of all kinds of urban wet weather
NH4 Cl 15.9
flows, including runoff from airports, parking lots, agricultural Peptone 30.6
runoff, combined sewer overflows and stormwater in general MgHPO·3H2 O 22.1
(Revitt et al., 2001; Debo and Reese, 2003; Fink and Mitsch, KH2 PO4 19.9
2005). Surface flow wetlands (Revitt et al., 2001; Greenway et FeSO4 ·7H2 O 4.9
al., 2006) and horizontal subsurface flow wetlands (Revitt et Starch 198.6
Milk powder 198.6
al., 2001, 2004; Shutes et al., 1999, 1997; Geary et al., 2006) have
Yeast 91.3
both been used for the treatment of urban wet weather flows.
Cr(NO3 )3 ·9H2 O 0.49
Emerging research also focuses on the use of vertical flow con- CuCl2 ·2H2 O 1.13
structed wetlands for the treatment of CSOs (Dittmer et al., MnSO4 ·H2 O 0.07
2005; Frechen et al., 2006; Welker, 2006). NiSO4 ·6H2 O 0.22
The removal of pollutants from water in constructed PbCl2 0.45
wetlands is achieved by an interactive combination of phys- ZnCl2 0.57
CdCl2 ·2H2 O 0.05
ical, chemical and biological processes as determined by
the physico-chemical conditions of the wetlands. Wong and
Geiger (1997) suggested that the k–C* model could be adapted
for the description of pollutant removal in wetlands treating filled with gravel to a height of 0.35 m; an inlet and outlet zone
stormwater. They suggested that the parameters in the k–C* was created with coarse gravel (16–32 mm) having a porosity (ε)
model would differ between wetlands used for treatment of of 0.43; the remainder of the beds was filled with finer gravel
wastewater and stormwater treatment due to the unsteady, (3–8 mm; ε = 0.38). Water levels during the experiments were
intermittent nature of the inflow of stormwater. Carleton et al. kept at a height of 0.35 m in the SSFCWs. Both types of wet-
(2001) investigated data from 35 studies on 45 wetlands used lands were planted with seedlings of common reed (Phragmites
for stormwater treatment and found that first-order removal australis) at a plant density of 25 plants m−2 (the SSFCWs in
rate constants for total phosphorus, ammonia and nitrate January 2005, the SFCWs in July 2006). The aboveground plant
were similar to values reported in the literature by Kadlec and parts of the SSFCWs were removed on a regular basis from the
Knight (1996) for wastewater treatment wetlands. Dittmer et wetland. Coverage for both types of wetland was over 90% at
al. (2005) described how a model, developed for the simula- the time of the experiments. However, the reed plants in the
tion of the treatment of municipal wastewater could be used SSFCWs formed more shoots and the shoots were stronger
to describe the removal of pollutants in a vertical flow con- and thicker. Shoot length in both wetland types was between
structed wetland used for treatment of CSOs in spite of the 1.2 and 1.5 m.
stochastic nature of the loading and loading regime of the The experiments were carried out from September to
incoming water. Based upon existing data, models for domes- November 2006 and November 2006 to February 2007. The
tic waste water can also be used for the removal of pollutants mean temperature during the tests with the SFCWs and
in CSOs without large modifications. SSFCWs was, respectively, 8 and 14 ◦ C. No artificial illumina-
The use of constructed wetlands represents a relatively tion was provided during this period. The experiments would
new approach for stormwater treatment (Dittmer et al., 2005; ideally have been conducted in the spring or summer but
Mungasavalli and Viraraghavan, 2006; Line et al., 2008). In this timing was influenced by factors beyond the control of the
study, two types of pilot-scale constructed wetlands, surface authors. All systems were batch-fed with synthetic wastewa-
(SFCWs) and subsurface flow (SSFCWs), were assessed for CSO ter (modified from Boeije et al., 1998) (Table 1). Each simulated
treatment. Besides an evaluation of the performance of both overflow event consisted of three successive batches with dif-
wetland types, emphasis was put on the difference in physico- ferent theoretical concentrations of pollutants (C1 ; C2 = 1/2C1
chemical conditions in the wetlands and their influence on and C3 = 1/4C1 ), which were successively introduced into the
the removal of nitrogen, phosphorous and chemical oxygen constructed wetlands. Dilution of the synthetic wastewater
demand (COD). was done by tapwater. The diluted synthetic wastewater was
allowed to stand for 30 min under continuous stirring. The
tapwater had a NO3 –N concentration of 3.4 ± 0.1 mg NO3 –N
2. Materials and methods l−1 . The applied concentrations were based on concentrations
measured in a full-scale CSO in Bornem, Belgium. The vol-
Four pilot-scale constructed wetlands were set up in a green- ume of one batch was 76 ± −4l, an overflow event (i.e. three
house, i.e. two replicate SFCWs (2.4 m × 0.3 m × 0.6 m) and two successive batches) corresponded with 227 ± −6l. The three
replicate SSFCWs (2.5 m × 0.625 m × 0.50 m). The SFCWs were different concentrations simulated the typically decreasing
filled with a 10 cm thick soil layer and were filled with water pollutant concentrations that enter a wetland during a sewer
to a height of 40 cm (water height of 30 cm). The SSFCWs were overflow event. In the SFCWs three tests with an overflow
 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183
event were carried out, in the SSFCWs two tests were per-
formed. In between two experiments the wetlands were fed
with tapwater to prevent drying-up. Contrary to the SSFCWs,
the water in the SFCWs was drained before introducing a
new overflow. The water present in the SSFCWs was forced
out of the wetlands when a new overflow was added. As such
the added water replaced the water that was still present
in the SSFCWs. As tapwater was used in the experiments,
the effect of chlorine present in the tapwater on the biofilm
needs to be evaluated. The available chlorine will react with
the organic material and biofilm present in the reedbeds, but
only a small part of it will be oxidised by it due to excess of
organic material present in the wetlands. Furthermore, if any
negative effect was present, this would be limited to the inlet
zone of the wetlands. As such, we concluded that is was safe
to use tapwater for dilution and irrigation.
Water samples were taken from each waste water batch
before application in the constructed wetlands. In the beds
themselves, water samples were collected at different time
intervals (day 0, 1, 3, 6 and 8) and at various distances from
the inlet (SFCWs: 0.3; 0.95; 1.55 and 2.2 m—SSFCWs: 0.2; 0.8;
1.4 and 2.3 m). For the SSFCWs an additional sampling event
after 12 h was included to evaluate the short-term removal.
The time interval between application of the first batch and
collection of the samples on day 0 was approximately 4 h. In
the SFCWs water samples were taken at a depth of 15 cm below
the water surface. For the SSFCWs water samples were taken
at depths of 15 and 30 cm. Water samples were taken with a
silicone tube and syringe and the samples from both depths
taken in the SSFCWs were pooled into one combined sample
for further processing.
Determination of NH4 –N was performed immediately after
sampling. To preserve samples for Ntot analysis, H2 SO4 was
added to a pH lower than 2. Both NH4 –N and Ntot were
determined by distillation following procedures described in
Standard methods for the examination of water and waste
water (Eaton, 1995). Nitrate-concentrations were determined
immediately by means of anion chromatography (Chromato-
graph 761 and Metrosep A-column, Methrohm, Switzerland).
Total phosphorus content was determined by colorimetry
using a Jenway 6500 spectrophotometer (Jenway, Essex, UK)
after digestion with potassium persulphate as described by
Eaton (1995). The redox potential was measured with com-
bined platina and gel reference electrodes (HI 3090 B/5; HI 9025,
Hanna Instruments, Ann Arbor, USA) that were permanently
present in the constructed wetlands at a depth of 0.15 m in the
water column and at the following distances from the inlet:
0.3; 0.95; 1.55 and 2.2 m for SFCWs, and 0.2; 0.8; 1.4 and 2.3 m
for SSFCWs. Chemical oxygen demand (COD) was determined
using Nanocolor test kits (Macherey-Nagel, Düren, Germany).
For overflow event 1 in the SSFCWs and event 2 in the SFCWs,
water samples were filtered over 0.45 ␮m pore size filters. This
allowed partitioning total COD (CODtot, measured on unfil-
tered samples) into soluble COD (CODsol, 0.45 ␮m filtered)
and particulate COD (CODpart, calculated as the difference
between CODtot and CODsol).
All statistical analyses were done with SPSS 15.0 (SPSS,
Chicago, USA). Two-sample t-tests were used to evaluate the
significance of differences between the two types of con-
structed wetlands and between the replicates. A confidence
177
level of 5% was adapted to evaluate the significance in the
different tests.
For plotting concentration profiles as a function of time
concentrations obtained for the different locations along the
length of the constructed wetlands were pooled together. This
was done because of following assumption: when a new over-
flow event enters the wetland, the water that is still present
in the wetland will be mixed and forced out of the wetland
as a whole. As such the mean concentration of pollutants
present at that time in the wetland will be the only relevant
concentration.
Removal efficiencies were calculated based on the amount
of a certain element that was introduced to the wetlands and
the amount that was recovered after 8 days. Concentration
profiles describing the removal of the pollutants as a function
of time in both wetland types were fitted to the first-order k–C*
model (Kadlec and Knight, 1996):
Ct − C∗
= e−kv t
C0 − C∗
where Ct the average concentration of the pollutant at time
t (mg l−1 ), C* the average background concentration of the pol-
lutant (mg l−1 ), C0 the initial concentration of the pollutant
(mg l−1 ), kv the volumetric removal rate constant (d−1 ), and t
the treatment time (d).
First-order rate removal constants and background concen-
trations were obtained for the removal of Ntot and CODtot for
both constructed wetland types. To differentiate between rate
constants for ‘fast’ and ‘slow’ removal, influent concentrations
were either included or excluded. The kv -value obtained by
excluding the influent concentrations was set as kv slow . kv fast
was calculated as the difference between the kv value obtained
when including the influent concentrations, and kv slow . Areal
removal rate constants (ka ) were also calculated as ka = kv εh
with h the water height (0.35 and 0.30 m for the SSFs and
SFCWs respectively) and the porosity ε (0.42 and 0.95 for the
SSFCWs and SFCWs, respectively).
3. Results
There were no significant differences between the two repli-
cates of each wetland type and this for all considered
pollutants (p < 0.05). After 1 day the redox potential in the
SSFCWs stabilised and remained stable for the further dura-
tion of the experiment. The redox potential increased towards
the outlet zone from −200 to −70 mV whereas for the SFCWs
no differences were observed along the length profile of the
beds. In the SFCWs the redox potential showed a less rapid
decline, but reached a similar value (−232 ± 35 mV) after day
3. The redox potential increased up to − 99 ± 20 mV on day 8.
Removal of Ntot occurred faster in the SSFCWs than in the
SFCWs (Fig. 1) with Ntot in the SSFCWs decreasing to con-
centrations of 0.90 ± 0.44 mg l−1 Ntot after 8 days. The average
removal efficiency for Ntot in the SSFCWs and SFCWs was
96.7 ± 1.9% and 36.6 ± 3.3% respectively (Table 2). Although
the global N-removal in the SFCWs was low, nitrogen forms
with time shifted from mainly organic and NO3 –N to domi-
nantly NH4 –N (Fig. 2). The NH4 –N supplied with the influent
was not detected in the SSFCWs, indicating a fast removal
178 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183

Fig. 2 – Fractionation of the different N-forms

(average ± 1S.D.) in the water of the SFCWs during overflow
3 as a function of time.

A substantial part of P was removed within the first 4 h

of the experiment in both types of constructed wetlands
(Fig. 1). Adsorption and precipitation are considered as the
main removal mechanisms for P. Overall removal efficien-
cies were much higher for the SSFCWs (Table 2). For both the
SFCWs and the SSFCWs a gradual release of P was observed
with time. Calculating release rates based upon the surfaces
of the beds (SSFCWs: 1.56 m2 ; SFCWs: 0.72 m2 ) and a given
retention time of 8 days showed values of 9 mg P m−2 d−1 and
14 mg P m−2 d−1 for the SSFCWs and SFCWs, respectively.
Fig. 1 – Removal patterns of Ntot, P and CODtot as a CODtot removal occurred faster (p < 0.005) in the SSFCWs
function of time for the SSFCWs (dashed line) and SFCWs and was removed to a higher extent (Fig. 1) although the influ-
(full line). Mean inlet concentrations are represented by ent concentrations were similar. The C1 batch had a CODtot
open symbols. concentration of 434 ± 50 and 450 ± 30 mg O2 l−1 , respectively
in the SFCWs and SSFCWs. After 4 h the CODtot concentra-
tion in the SSFCWs was half the concentration measured in
the SFCWs (107 ± 20 mg O2 l−1 vs. 214 ± 8 mg O2 l−1 ). The con-
centration at the end of the experiment was 95 ± 4 mg O2 l−1
occurring mostly within the first 24 h. In the SSFCWs con-
and 21 ± 15 mg O2 l−1 for the SFCWs and SSFCWs, respectively.
centrations dropped from 2.5 ± 2.2 mg l−1 NH4 –N present in
For each wetland type, a fractionation of the CODtot for one
the influent to less than 0.4 mg l−1 (data not shown) and no
overflow event (SSFCWs: event 1; SFCWs: event 3) is pre-
increase of NH4 –N was found during the further duration of
sented in Table 3. Although the composition and fractionation
the experiment. In contrast to the SSFCWs, increasing NH4 –N
were equal for both overflows on the SSFCWs and SFCWs,
concentrations were observed with time for the SFCWs:
the concentration of CODpart was higher in the SFCWs 4 h
NH4 –N concentrations increased from 2.7 ± 0.4 mg l−1 on day
after application (day 0). CODtot was removed much faster in
0 to 18.1 ± 0.2 mg l−1 on day 8. The influent concentration
the SSFCWs than in the SFCWs, as evidenced by the differ-
of nitrates was 8 ± 1.5 mg NO3 –N l−1 throughout the experi-
ence between CODtot in the influent and that on day 0. This
ments. Concentrations of NO3 –N after 1 day were lower than
could be mainly attributed to the faster removal of CODpart in
0.07 and 0.2 mg NO3 –N l−1 for the SSFCWs and SFCWs, respec-
the SSFCWs. The CODpart was for each sampling time signif-
tively.
icantly higher in the SFCWs. The overflow caused an increase
of CODpart in the SFCWs probably due to resuspension of the
sediment when the water entered the system. Moreover, the
Table 2 – Removal efficiencies (mass-based) for the CODsol was lower in the SSFCWs than in the SFCWs. The
different pollutants in the SFCWs and SSFCWs after 8 overall removal efficiency was 88 ± 4% for the SSFCWs and
days. 61 ± 7% for the SFCWs (Table 2). For the SFCWs the removal
Pollutant SFCW (%) SSFCW (%) efficiency decreased systematically over the overflow events
from 67.8 ± 2% for overflow 1 to 52 ± 0.1% for overflow 3. The
Ntot 36.6 ± 3.3 96.7 ± 1.9 COD-removal in the SSFCWs remained constant throughout
P 36.0 ± 5.0 71.1 ± 7.7
the different overflow events.
NO3 –N 99.7 ± 0.1 99.1 ± 0.5
CODtot 60.8 ± 7.1 88.1 ± 3.5
Initial contaminant removal was faster in the SSFCWs than
in the SFCWs. After 12 h the concentration of NO3 –N was
 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183 179

Table 3 – Fractionation of CODtot (average ± 1S.D.) for the SSFCWs (overflow 1) and SFCWs (overflow 3).
CODtot CODpart CODsol

SFCW SSFCW SFCW SSFCW SFCW SSFCW

C1 434 ± 50 450 ± 30 43 ± 15 151 ± 35 361 ± 29 300 ± 5

C2 212 ± 25 227 ± 14 15 ± 18 21 ± 39 195 ± 22 205 ± 27
C3 102 ± 10 113 ± 7 7 ± 16 4±2 101 ± 14 109 ± 7
Day 0 230 ± 27 96 ± 14 50 ± 24 19 ± 8 179 ± 48 76 ± 17
Day 1 137 ± 16 50 ± 5 37 ± 10 6±4 100 ± 9 45 ± 7
Day 3 113 ± 15 20 ± 10 54 ± 15 2±4 59 ± 2 18 ± 6
Day 6 73 ± 11 16 ± 1 12 ± 11 <1 61 ± 5 <15
Day 8 n.d. 16 ± 1 n.d. <1 n.d <15

n.d.: not determined.

Table 4 – Short-term removal of Ntot, NO3 –N and P after 4, 12 and 24 h for the SSFCWs (average ± 1S.D.).
Influent 4h 12 h 24 h

Ntot (mg l−1 ) 31.9 ± 21.1 19.4 ± 5.4 (39) 12.1 ± 2.6 (62) 4.9 ± 1.3 (85)
NO3 –N (mg l−1 ) 10.2 ± 5.9 7.0 ± 0.7 (32) 0.9 ± 1.6 (91) 0.04 ± 0.03 (>99)
P (mg l−1 ) 4.0 ± 1.9 1.00 ± 0.21 (75) 1.03 ± 0.77 (74) 1.25 ± 0.85 (67)

Removal percentages (%) are shown between brackets.

Table 5 – Coefficients for the k–C*-model assessed with (+ influent) or without (− influent) influent concentrations.
R C* (mg l−1 ) kv (d−1 ) ka (m d−1 )
Ntot SFCW + Influent 0.98 18.7 ± 0.5 6.0 ± 1.6 1.7 ± 0.5
− Influent 0.98 18.3 ± 0.3 1.2 ± 0.4 0.3 ± 0.1
SSFCW + Influent 0.99 1.5 ± 0.6 2.8 ± 0.4 0.4 ± 0.1
− Influent 0.99 1.1 ± 0.1 1.9 ± 0.1 0.29 ± 0.02

CODtot SFCW + Influent 0.99 95.9 ± 10.2 0.6 ± 0.2 0.17 ± 0.06
− Influent 0.99 89.6 ± 6.9 0.4 ± 0.1 0.11 ± 0.03
SSFCW + Influent 0.99 36.7 ± 9.5 7.0 ± 1.8 1.1 ± 0.3
− Influent 0.99 24.3 ± 2.9 0.8 ± 0.1 0.12 ± 0.02

Table 6 – First-order volumetric and area-based (subscripts v and a, respectively) removal rate constants determined for
fast and slow removal in both the SFCWs and SSFCWs.
kv fast (d−1 ) kv slow (d−1 ) ka fast (m d−1 ) ka slow (m d−1 )
Ntot SFCW 4.8 ± 2.0 1.2 ± 0.4 1.4 ± 0.6 0.3 ± 0.1
SSFCW 0.9 ± 0.5 1.9 ± 0.1 0.14 ± 0.07 0.54 ± 0.03

CODtot SFCW 0.2 ± 0.3 0.4 ± 0.1 0.06 ± 0.09 0.11 ± 0.03
SSFCW 6.2 ± 1.9 0.8 ± 0.1 0.9 ± 0.3 0.12 ± 0.02

less than 1 mg l−1 (Table 4). In first instance removal of P was
observed but already after 12 h the concentrations of P in
the water started to increase. The main removal of P (75%)
occurred within the first 4 h after the overflow event. This cor-
responded with the sharp decline of the redox potential from
173 ± 107 mV after 4 h to −192 ± 54 mV after 24 h.
A considerable amount of contaminants was already
removed 4 h after application of the simulated overflow
event, suggesting high rate removal processes. During the
further duration of the experiment a slower removal was
observed. First-rate removal constants for Ntot and CODtot
were calculated with inclusion and exclusion of the influent
concentration data. The calculated constants are presented in
Table 5. The inclusion of influent concentrations resulted in
higher C* and k-values. Deriving kfast and kslow showed that
fast occurring reactions were important for the removal of
Ntot in the SFCWs and the removal of CODtot in the SSFCWs
(Table 6).
4. Discussion
The SSFCWs were planted in January 2005, whereas the SFCWs
were planted in July 2006. At the start of the experiments both
180 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183
wetland types had an actual age of 15 months (SSFCWs) and
5 months (SFCWs). The high plant density that was applied
to the wetlands (25 plants m−2 ) results in a faster maturation
process of the vegetation compared to lower plant densities
(USEPA, 2000). Another factor at which both wetlands may
differ is the release of components during degradation of
dead plant material. In the SFCWs, as it was recently planted,
no decaying plant material was present. For the SSFCWs the
standing biomass was removed in the past and as such no
litter build-up was present on top of both wetland types.
Microbial removal of pollutants is a very important pathway
for several contaminants. Because of the difference in age
between both wetland types, a difference in biofilm develop-
ment may be present. Development of microbial biomass in
the media of a SSFCW typically requires three to six months
(USEPA, 2000). Tao et al. (2006) investigated the maturation of
biofilms in SFCWs used for woodwaste leachate treatment. It
took <1–6 weeks for maturation of the biofilm on submerged
plant surfaces and the sedimentary microbial community,
with longer maturation time needed for the attached micro-
bial communities than for the suspended communities.
Although the SFCWs were much younger, it can be expected
that in both used wetland types, a stable microbial commu-
nity was present and that gravel, submerged plant parts, soil
surface and the water column were completely colonised by
biofilms.
N-removal in the SFCWs was very limited. Removal of
nitrogen is influenced by temperature (Gardner et al., 2000;
Wang et al., 2008). Absence of total Kjeldahl nitrogen removal
was observed in a constructed wetland by Reuter et al. (1992).
They attributed this to reduced nitrification and ammonifi-
cation rates, which was associated with a poor vegetation
growth. Although the reed plants in the SFCWs had been accli-
matised during more than 5 months, the lower temperatures
in the greenhouse during the test period resulted in a poor con-
dition of the reed. Below 15 ◦ C neither the micro-organisms
responsible for nitrogen removal nor uptake by vegetation
functions properly (Vymazal, 1999; Akratos and Tsihrintzis,
2007).
After 8 days, almost all the N was present in the form
of NH4 –N within the SFCWs, indicating that ammonification
occurred (Fig. 2). Ammonification occurs both under aerobic
and anaerobic conditions (Vymazal, 2007). Part of the Ntot
present in the influent consisted of nitrogen associated with
particulate matter. This part was quickly removed by sedimen-
tation. The compounds that were added to act as a carbon
source, such as starch, milk powder and peptone, did not dis-
solve completely and contained also N, acting as a source of
particulate N. The decrease of Ntot that was observed between
the addition of the combined sewer overflow and 4 h later
was attributed to sedimentation, sorption and nitrate-removal
reactions. The nitrates were efficiently removed after 4 h by
denitrification for both the SSFCWs and SFCWs and no sig-
nificant levels of nitrates were detected later on. Such strong
denitrification activity at redoxpotentials lower than 300 mV
was clearly shown in a microcosm incubation experiment
by Yu et al. (2007). The nitrates present in the influent were
not detected in the SSFCWs after 4 h, pointing out the quick
removal by plant uptake or turn-over to intermediate products
of the denitrification and N2 .
The consistent low nitrate concentrations in both wet-
land types during the further duration of the experiment
indicated limiting conditions for nitrification due to very low
dissolved oxygen levels in the water. Some authors believe
that the amount of oxygen escaping from the plant roots into
the anaerobic environment is very limited, and atmospheric
oxygen diffusion is therefore the main source of oxygen in
wetlands (Armstrong and Armstrong, 1988; Brix, 1993; Kadlec
and Knight, 1996). One can assume however that oxygen
supply due to atmospheric diffusion is negligible in SSFCWs
because the water flows between the gravel and only limited
contact is possible between water and atmosphere. Then, root
oxygen loss would constitute the main oxygen source. How-
ever Wu et al. (2001) found in a comparative study between
SSFCWs and SFCWs that the atmospheric oxygen-diffusion
rates of SSFCWs were 4–17 times higher than those of SFCWs.
Wu et al. (2001) concluded their study with the suggestion that
the amount of oxygen transferred through both the roots and
atmospheric diffusion could not fully explain the high removal
of NH4 in the SSFCWs and SFCWs under study by nitrification
and denitrification reactions.
Authors have stated that constructed wetlands are char-
acterized by the occurrence of both aerobic and anaerobic
microsites within the wetland, allowing both aerobic and
anaerobic reactions to occur (Vymazal, 1999). The overall good
removal efficiency of Ntot within the SSFCWs seems to be in
accordance with the assumption that nitrification could occur
near the roots after which diffusion of the formed nitrate to the
anaerobic surroundings occurred, followed by denitrification.
Decreasing nitrogen concentrations can go together with-
out significant increase of nitrate or nitrite as was observed
by Akratos and Tsihrintzis (2007). Denitrification is primarily
determined by the rate of diffusion of the nitrates to zones
where denitrification might take place (Flynn et al., 1999).
After 4 h no significant removal of NH4 –N was observed
in the SFCWs (Fig. 2). Thus, N-removal by sorption of NH4 –N
was negligible within the first 4 h for the SFCWs, compared
to the high contribution of this process in the SSFCWs where
no NH4 could be detected 4 h after application of the over-
flow event. Adsorption onto the substrate may be of more
importance in SSFCWs because of the larger surface area pro-
vided by the gravel (Sikora et al., 1995). During a previous
experiment on the same SSFCWs as used during this study
maximum sorption capacity of the gravel was determined at
17 mg NH4 –N kg−1 gravel (Torres, 2005). In agreement with our
results, Wu et al. (2001) found higher ammonium removal effi-
ciencies in SSFCWs than in SFCWs. Depending on the pH of the
water, conversion of NH4 –N to NH3 followed by volatilisation
might become important. As there is a closer contact between
the water and air in the SFCWs, this reaction may, at higher
pH, be more important in SFCWs (Vymazal, 2007). Connolly et
al. (2004) described how the first step of ammonium-removal
in a vertical flow constructed wetland was adsorption of NH4 +
onto the media, followed by nitrification and denitrification
reactions. Sorption of NH4 –N is enhanced in more reducing
conditions, stimulating the removal by sorption in the SSFCWs
(Sikora et al., 1995).
The sorption of NH4 + onto the substrate of the SSFCWs
can be followed by a gradual release. The released NH4 –N can
be removed by plant uptake as plants prefer the uptake of
 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183
the more reduced NH4 + –N to the uptake of NO3 − (Kadlec and
Knight, 1996). This release could have occurred between two
monitoring campaigns, when tap water was used for irrigating
the beds. Nutrient-limiting conditions between two overflow
events may have enhanced desorption so that no saturation
of the substrate occurred and adsorption sites were readily
available when a new overflow event entered the system.
Evaluating the importance of plant uptake in the SFCWs
and SSFCWs one can conclude that plant uptake will be of
higher importance in the SSFCWs. This was mainly due to
the better condition of the reed plants in the SSFCWs during
the test period, but might also depend on the better con-
tact between plant roots and water in the SSFCWs. Next to
nutrient uptake from the water, also nutrients present in the
soil of the SFCW are available for plant uptake. Recently, the
presence of alternative NH4 -removal pathways in constructed
wetlands has been brought under attention (Sun and Austin,
2007; Vymazal, 2007). Autotrofic denitrification, better known
as the ANAMMOX-process, could also have contributed next to
plant uptake and adsorption onto the substrate to the removal
of nitrogen. In this process nitrite and ammonium are directly
transformed into nitrogen gas (Mulder et al., 1995).
As such, we attribute the difference in removal between
the SSFCWs and SFCWs to a combination of characteristics
intrinsic to the type of system and lower temperatures dur-
ing the testing period with the SFCWs (8 ◦ C) affecting both
plant growth and microbial processes. Sorption of NH4 onto
the gravel was identified as the most probable removal mech-
anism in the SSFCWs.
The gravel in the SSFCWs was more important in removal
of P than the soil substrate in the SFCWs. The gravel in the
SSFCWs has a higher adsorption capacity than the soil and
provides a larger contact area as water flows within the gravel
substrate rather than on top of the soil surface. Many studies
demonstrated P removal for both SSFCWs and SFCWs (Green
and Upton, 1993; Kadlec and Knight, 1996; Drizo et al., 2000;
Braskerud, 2002). In a column-experiment simulating SFCWs
Dunne et al. (2005) observed an initial release of P, followed by
a decrease. This release resulted in a concentration increase
of 0.4 mg l−1 after 2 days. Over the duration of this study, P in
the SFCWs increased with approximately 0.4 mg l−1 although
the initial P-concentrations present in the soil were 2–4 times
higher (1800 mg P kg−1 soil) than the soils that were used by
Dunne et al. (2005). The increase of P in the SSFCWs was a lit-
tle higher (0.6 mg l−1 ). The release of oxygen by plant roots may
increase the adsorption capacity of wetlands for P (Walhugala
et al., 1987). The release of P coincided with decreasing redox
potentials in both wetland types. Redox conditions strongly
influence the chemistry of P through indirect effects on the
solubility of Fe and Mn minerals. In addition, the associated
changes in pH can affect the solubility of Ca-bound phos-
phate and the sorption reactions involving variably charged
surfaces (Scalenghe et al., 2002). Reduction reactions in the
environment result in a net alkaline reaction, which tends to
be counteracted by the precipitation of carbonates of Fe(II) and
Mn(II) and the production of CO2 and organic acids originat-
ing from decomposing organic material (Yu et al., 2007). Gu
and Dreschel (2008) found a correlation between the removal
of Ca, present in the influent water, and the removal of P. The
removal of P is less influenced by the loading rate than by the
181
mean retention time. In contrast with Carleton et al. (2001),
who found that an increase of the retention time lowered the
effect of resuspension of the P sorbed or associated with the
sediment, our experiment showed for both the SSFCWs and
SFCWs an initial removal of P followed by a gradual release of
the sorbed P. The current experiment showed the importance
of the retention time when release of P from the substrate
is presumably to occur. Shorter retention times would have
resulted in higher removal efficiencies. Also for the removal of
P, influence of temperature has been mentioned in the litera-
ture (Wang et al., 2008), but this was described as an indirect
effect. Lower temperatures are associated with winter and this
period goes together with decomposition of the litter, resulting
in P-release.
COD removal occurred significantly faster in the SSFCWs
than in the SFCWs (p < 0.001) as a combination of enhanced
sedimentation, the presence of a higher contact surface with
the biofilm and probably a more balanced assembly of biofilm
in the SSFCWs as they were already in use for a longer time.
The gravel in the SSFCWs enhanced sedimentation of COD-
part as could be subdued from the rapid decrease of CODpart.
The presence of the gravel has a dual action. It enhances
sedimentation by slowing down the water current, and sec-
ondly, the biofilm, attached to the gravel will enhance the
trapping of particulates. The gravel also prevents resuspen-
sion to occur, which was clearly observed in the SFCW. As
water from combined sewer overflows is highly loaded with
particulate material and pollutants tend to be associated with
the particulate fraction, an effective removal of this fraction
may strongly increase removal efficiencies of such systems.
However, the capacity of removing particulate material may
change over time resulting in a decrease of the retention of
suspended solids (Tanner et al., 1998). SSFCWs tend to allow
a better removal of particulate material due to the filtering
effect of the gravel, although clogging can occur, resulting in
lower removal performances. Kadlec and Knight (1996) stated
that the effect of temperature on the removal of BOD for
both SSFCWs and SFCWs was negligible. Also the final COD-
sol concentration was lower in the SSFCW than in the SFCW.
Higher concentrations of attached biofilm may have enhanced
the removal of soluble organic compounds. Stein et al. (2006)
observed a substantial COD removal almost immediately upon
filling during a batch-loaded column experiment simulating
SSFCWs. Residual levels (C*) were reached by days 3–14 and
sometimes COD-values approached C* within 1 day.
For all the investigated parameters, the SSFCWs performed
better, resulting in a faster removal down to lower concentra-
tions. This study confirms the results of Shutes (2001), which
indicated that SSFCWs are more efficient than SFCW when
removing pollutants at higher application rates (Shutes, 2001).
Due to higher removal rates, shorter retention times can be
applied. The main drawback for both systems is the large area
needed for construction and the need for a constant standing
volume of water to preserve the vegetation. For buffering a vol-
ume of 1000 m3 and working with a total height of 0.5 m (0.4 m
available for storage and a remaining water height of 0.1 m)
the required surface is 0.65 ha for the SSFCWs and 0.25 ha
for the SFCWs. Also other pollutants than the ones discussed
here can be of major concern in stormwater and CSO treat-
ment. Metal removal and behaviour may differ between both
182 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183
wetland types. Furthermore, performances observed in full-
scale systems may differ from those in small-scale systems.
SSFCWs seem to be usable when CSO- and stormwater treat-
ment can be combined with tertiary treatment of wastewater.
The application of this type of treatment without secondary
water inputs and to guarantee at the same time a satisfac-
tory plant growth might prove to be quite difficult as the
performance will be highly dependent on the frequency of
overflowing. SSFCWs are sensitive to drying-up, resulting in
plant death, when no regular water input is present. This is
important, especially during summer when high evapotran-
spiration rates occur. However, peak concentrations can be
removed more quickly by SSFCWs, resulting in concentrations
that can be discharged earlier compared to SFCWs. SFCWs can
store a larger volume of water on the same surface area than
SSFCWs and therefore a higher volume of water remaining in
the bed can be used without losing the needed buffer capacity.
Therefore SFCWs seem to be more applicable as stand-alone
constructed wetlands in the field without the need for sec-
ondary water inputs and when the frequency of overflows is
rather low. When surface area is scarce, when dealing with
highly frequent overflow events or rather high concentrations,
preference is given to SSFCWs.
5. Conclusion
Nitrogen, P and COD present in synthetic wastewater simulat-
ing combined sewer overflow events were removed faster by
SSFCWs than SFCWs. In general the end concentrations of the
investigated pollutants were lower in the SSFCWs. Removal
of nitrogen was strongly inhibited in the SFCW. The very
high removal efficiencies of Ntot obtained in the SSFCWs are
attributed to a combination of efficient ammonification, plant
uptake and effective sorption of the formed NH4 –N to the
substrate. Removal by nitrification and denitrification upon
desorption of the ammonium occurred probably in the time
span between two testing periods. In both wetland types an
initial removal of P was followed by a release within the first
day. The quick removal of P was due to sorption. More reduc-
ing conditions resulted in a gradual release of the sorbed P.
Besides removal performances practical design issues such as
the surface area needed to store and treat a certain amount of
stormwater or CSOs, dependence of other water inputs (and
the possible presence of these inputs in combination with the
frequency of the overflow events) need to be considered when
choosing a wetland type for treatment of combined sewer
overflows.
references
Akratos, C.S., Tsihrintzis, V.A., 2007. Effect of temperature, HRT,
vegetation and porous media on removal efficiency of
pilot-scale horizontal subsurface flow constructed wetlands.
Ecol. Eng. 29, 173–191.
Armstrong, J., Armstrong, W., 1988. Phragmites australis—a
preliminary study of soil oxidizing sites and internal gas
transport pathways. New Phytol. 108, 373.
Boeije, G., Corstanje, R., Rottiers, A., Schowanek, D., 1998.
Adaptation of the CAS test system and synthetic sewage for
biological nutrient removal. Part I. Development of a new
synthetic sewage. Chemosphere 38, 699–709.
Braskerud, B.C., 2002. Factors affecting phosphorus retention in
small constructed wetlands treating agricultural nonpoint
source pollution. Ecol. Eng. 1, 41–61.
Brix, H., 1993. Macrophyte mediated oxygen transfer in wetlands:
transport mechanisms and rates. In: Moshiri, G.A. (Ed.),
Constructed Wetlands for Water Quality Improvement. CRC
Press LLC, Boca Raton, FL, p. 391.
Carleton, J.N., Grizzard, T.J., Godrej, A.N., Post, H.E., 2001. Factors
affecting the performance of stormwater treatment wetlands.
Water Res. 35, 1552–1562.
Connolly, R., Zhao, Y.Q., Sun, G.Z., Allen, S., 2004. Removal of
ammoniacal-nitrogen from an artificial landfill leachate in
downflow reed beds. Process Biochem. 39, 1971–
1976.
Debo, T.N., Reese, A.J., 2003. Municipal Stormwater Management.
CRC Press LLC, Boca Raton, FL, p. 1141.
Dittmer, U., Meyer, D., Langergraber, G., 2005. Simulation of a
subsurface vertical flow constructed wetland for CSO
treatment. Water Sci. Technol. 51, 225–232.
Drizo, A., Frost, C.A., Grace, J., Smith, K.A., 2000. Phosphate and
ammonium distribution in a pilot-scale constructed wetland
with horizontal subsurface flow using shale as a substrate.
Water Res. 39, 2483–2490.
Dunne, E.J., Culleton, N., O’Donovan, G., Harrington, R.,
Daly, K., 2005. Phosphorus retention and sorption by
constructed wetland soils in Southeast Ireland. Water Res. 39,
4355–4362.
Eaton, A.D., 1995. In: Eaton, A.D., Clesceri, L.S., Greenberg, A.E.
(Eds.), 1995. Standard Methods for the Examination of Water
and Waste water, 19th ed. American Public Health
Organisation, Washington, DC, USA, p. 1325.
Fink, D.F., Mitsch, W.J., 2005. Seasonal and storm event nutrient
removal by a created wetland in an agricultural watershed.
Ecol. Eng. 23, 313–325.
Flynn, N., Gardner, P.J., Maltby, E., 1999. The measurement and
analysis of denitrification rates obtained using soil
columns from river marginal wetlands. Soil Use Manage. 15,
150–156.
Frechen, F.B., Schier, W., Felmeden, J., 2006. The plant-covered
retention soil filter (RSF): the mechanical and biological
combined sewer overflow (CSO) treatment plant. Eng. Life Sci.
6, 74–79.
Gardner, P.J., Flynn, N., Maltby, E., 2000. A temperature correction
procedure for field-based soil denitrification experiments. Soil
Use Manage. 16, 260–263.
Geary, P.M., Mendez, H., Dunstan, R.H., 2006. Design
considerations in the performance of stormwater devices
incorporating constructed wetlands (1833–1842). In:
Proceedings of the 10th International Conference on Wetland
Systems for Water Pollution Control, September 23–29, Lisbon,
Portugal.
Green, M.B., Upton, J., 1993. Reed bed treatment for small
communities: UK experience. In: Moshiri, G.A. (Ed.),
Constructed Wetlands for Water Quality Improvement. CRC
Press, Inc., Boca Raton, FL, pp. 517–524.
Greenway, M., Jenkins, G., Polson, C., 2006. Macrophyte zonation
in stormwater wetlands—getting it right! (1807–1819). In:
Proceedings of the 10th International Conference on Wetland
Systems for Water Pollution Control, September 23–29, Lisbon,
Portugal.
Gu, B., Dreschel, T., 2008. Effects of plant community and
phosphorus loading rate on constructed wetland performance
in Florida, USA. Wetlands 28, 81–91.
Heaney, J.P., Wright, L., Sample, D., 1999. Research needs in urban
wet weather flows. Water Environ. Res. 71, 2.
Kadlec, R.H., Knight, R.L., 1996. Treatment Wetlands. Lewis
Publishers, Boca Raton, FL, p. 928.
 e c o l o g i c a l e n g i n e e r i n g 3 5 ( 2 0 0 9 ) 175–183
Line, D.E., Jennings, G.D., Shaffer, M.B., Calabria, J., Hunt, W.F.,
2008. Evaluating the effectiveness of two stormwater
wetlands in North Carolina. Trans. ASABE 51, 521–528.
Mulder, A., van de graaf, A.A., Robertson, L.A., Kuenen, J.G., 1995.
Anaerobic ammonium oxidation discovered in a denitrifying
fluidized bed reactor. FEMS Microbiol. Ecol. 16, 177–184.
Mungasavalli, D.P., Viraraghavan, T., 2006. Constructed wetlands
for stormwater management: a review. Fresenius Environ.
Bull. 15, 1363–1372.
Reuter, J.E., Djohan, T., Goldman, C.R., 1992. The use of wetlands
for nutrient removal from surface runoff in a cold climate
region of California—results from a newly constructed
wetland at Lake Tahoe. Environ. Manage. 36, 35–53.
Revitt, D.M., Shutes, R.B.E., Jones, R.H., Forshaw, M., Winter, B.,
2004. The performance of vegetative treatment systems for
highway runoff during wet and dry conditions. Sci. Total
Environ. 334–335, 264–270.
Revitt, D.M., Worall, P., Brewer, D., 2001. The integration of
constructed wetlands into a treatment system for airport
runoff. Water Sci. Technol. 44, 469–476.
Robson, M., Spence, K., Beech, L., 2005. Stream quality in a small
urbanised catchment. Sci. Total Environ. 357, 194–207.
Scalenghe, R., Edwards, A.C., Marsan, F.A., Barberis, E., 2002. The
effect of reducing conditions on the solubility of phosphorus
in a diverse range of European agricultural soils. Eur. J. Soil
Sci. 53, 439–447.
Scholz, M., Höhn, P., Minall, R., 2002. Mature experimental
constructed wetlands treating urban water receiving high
metal loads. Biotechnol. Prog. 18, 1257–1264.
Shutes, R.B.E., 2001. Artificial wetlands and water quality
improvement. Environ. Int. 26, 441–447.
Shutes, R.B.E., Revitt, D.M., Lagerberg, I.M., Barraud, V.C.E., 1999.
The design of vegetative constructed wetlands for the
treatment of highway runoff. Sci. Total Environ. 235, 189–197.
Shutes, R.B.E., Revitt, D.M., Mungur, A.S., Scholes, L.N.L., 1997.
The design of wetland systems for the treatment of urban
runoff. Water Sci. Technol. 35, 19–25.
Sikora, F.J., Zhu, T., Steinberg, S.L., Coonrod, H.S., 1995.
Ammonium removal in constructed wetlands with
recirculating subsurface flow: removal rates and mechanisms.
Water Sci. Technol. 32, 193–203.
Stein, O.R., Biederman, J.A., Hook, P.B., Allen, W.C., 2006. Plant
species and temperature effects on the k–C* first-order model
for COD removal in batch-loaded SSF wetlands. Ecol. Eng. 26,
100–112.
Suarez, J., Puertas, J., 2005. Determination of COD, BOD and
suspended solids loads during a combined sewer overflow
(CSO) event in some combined catchments in Spain. Ecol. Eng.
24, 201–219.
183
Sun, G., Austin, D., 2007. Completely autotrophic
nitrogen-removal over nitrite inlab-scale constructed
wetlands: evidence from a mass balance study. Chemosphere
68, 1120–1128.
Tanner, C.C., Sukias, J.P.S., Upsdell, M.P., 1998. Organic matter
accumulation during maturation of gravel-bed constructed
wetlands treating farm dairy waste waters. Water Res. 32,
3046–3054.
Tao, W., Hall, K.J., Duff, S.J.B., 2006. Performance evaluation and
effects of hydraulic retention time and mass loading rate on
treatment of woodwaste leachate in surface-flow constructed
wetlands. Ecol. Eng. 26, 252–265.
Torres, A.M.L., 2005. Subsurface flow constructed wetlands (SSF
CW): pilot-scale experiments and applicability in the
Philippines. MSc Environmental Sanitation (2004–2005), Ghent
University, 2005.
USEPA, 2000. Constructed wetlands treatment of municipal
wastewaters. Manual of EPA/625/R-99/010, U.S. Environmental
Protection Agency, Cincinnati, OH.
Vymazal, J., 1999. Nitrogen removal in constructed wetlands with
horizontal sub-surface flow—can we determine the key
process? In: Vymazal, J. (Ed.), Nutrient Cycling and Retention
in Natural and Constructed Wetlands. Backhuys Publishers,
Leiden, pp. 1–17.
Vymazal, J., 2007. Removal of nutrients in various types of
constructed wetlands. Sci. Total Environ. 380, 48–65.
Walhugala, A.G., Suzuki, T., Kurihara, Y., 1987. Removal of
nitrogen, phosphorus and COD from waste water using a sand
filtration system with Phragmites australis. Water Res. 21,
1217–1224.
Welker, A., 2006. Vertical flow constructed wetlands for enhanced
CSO treatment—an alternative for elimination of organic
pollutants? In: In Proceedings of the 10th International
Conference on Wetland Systems for Water Pollution Control,
vol. 693, September 23–29, Lisbon, Portugal.
Wang, Y., Inamori, R., Kong, H., Xu, K., Inamori, Y., Kondo, T.,
Zhang, J., 2008. Nitrous oxide emission from polyculture
constructed wetlands: effect of plant species. Environ. Pollut.
152, 351–360.
Wong, T.H.F., Geiger, W.F., 1997. Adaptation of waste water
surface flow wetland formulae for application in constructed
stormwater wetlands. Ecol. Eng. 9, 187–202.
Wu, M.-Y., Franz, E.H., Chen, S., 2001. Oxygen fluxes and
ammonia removal efficiencies in constructed treatment
wetlands. Water Environ. Res. 73, 661–666.
Yu, K., Böhme, F., Rinklelebe, J., Neue, H.-U., DeLaune, R.D., 2007.
Major biogeochemical processes in soils—a microcosm
incubation from reducing to oxidizing conditions. Soil Sci.
Soc. Am. 71 (4), 1406–1417.