Available online at www.sciencedirect.

com

Environmental Pollution 155 (2007) 125e131

www.elsevier.com/locate/envpol

Sulphur transformation and deposition in the rhizosphere of

Juncus effusus in a laboratory-scale constructed wetland
A. Wiessner, P. Kuschk*, M. Jechorek, H. Seidel, M. Kästner
Department of Bioremediation, Helmholtz Centre for Environmental Research e UFZ, Permoserstraße 15, 04318 Leipzig, Germany
Received 8 August 2007; received in revised form 17 October 2007; accepted 24 October 2007

In an experimental constructed wetland a clear correlation of the occurrence of reduced S-species with decreasing
C and N removal performance and plant viability was observed.

Abstract

Sulphur cycling and its correlation to removal processes under dynamic redox conditions in the rhizosphere of helophytes in treatment
wetlands are poorly understood. Therefore, long-term experiments were performed in laboratory-scale constructed wetlands treating artificial
domestic wastewater in order to investigate the dynamics of sulphur compounds, the responses of plants and nitrifying microorganisms under
carbon surplus conditions, and the generation of methane.
For carbon surplus conditions (carbon:sulphate of 2.8:1) sulphate reduction happened but was repressed, in contrast to unplanted filters
mentioned in literature.
Doubling the carbon load caused stable and efficient sulphate reduction, rising of pH, increasing enrichment of S2 and S0 in pore water, and
finally plant death and inhibition of nitrification by sulphide toxicity.
The data show a clear correlation of the occurrence of reduced S-species with decreasing C and N removal performance and plant viability in
the experimental constructed wetlands.
Ó 2007 Elsevier Ltd. All rights reserved.

Keywords: Ammonia; Constructed wetland; Juncus effusus; Laboratory-scale reactor; Sulphur cycle; Wastewater treatment

1. Introduction transformations, such as nitrification, denitrification, minerali-

The rhizosphere of constructed wetlands offers specific
macro and micro gradients of redox conditions enabling the
development of highly diverse microbial consortia capable
of different beneficial redox reactions (Bezbaruah and Zhang,
2004; Liesack et al., 2000; Wießner et al., 2005). The near root
environment was found to be of high importance for contam-
inant removal because of the capability of helophytes to
release oxygen and organic carbon at the same time into their
rhizosphere resulting in micro gradient redox conditions of
high spatial and temporal dynamics (Liesack et al., 2000;
Wei et al., 2003; Wießner et al., 2005). Different microbiological
* Corresponding author. Tel.: þ49 341 235 2821; fax: þ49 341 235 2492.
E-mail address: peter.kuschk@ufz.de (P. Kuschk).
zation of organic carbon, methanogenesis, reduction, and oxida-
tion of several sulphur compounds occur simultaneously on
a small spatial scale (Bezbaruah and Zhang, 2004; Choi et al.,
2006; Liesack et al., 2000; Wießner et al., 2005). The dynamics
of redox processes of sulphur cycling and its effectiveness is un-
known and their impact on other processes is underestimated
(Kalin, 2004; Lamers et al., 2002; O’Sullivan et al., 1999;
Wiessner et al., 2005; Wind and Conrad, 1997; Wind et al.,
1999).
Processes of the sulphur cycling are known to be determi-
native for high redox dynamics (Holmer and Storkholm,
2001; Lens et al., 1998; Wind and Conrad, 1997) and various
sulphur compounds are present in natural and constructed
treatment wetlands caused by natural and anthropogenic sup-
ply particularly in industrial wastewater and from mining

0269-7491/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.envpol.2007.10.027
126 A. Wiessner et al. / Environmental Pollution 155 (2007) 125e131
industries (Kalin, 2004; O’Sullivan et al., 1999; Vymazal and
Kröpfelová, 2005; Winter and Kickuth, 1989). The importance
particularly to treat sulphur-rich waters (acid mine drainages)
to remove acidity and/or heavy metals increased continuously.
However, for these special applications sulphate removal is
particularly in the focus but the interrelations of the processes
inside the rhizosphere are poorly understood (Kalin, 2004;
Morrison and Aplin, 2003; O’Sullivan et al., 2004; Webb
et al., 1998).
In many areas, considerable amounts of sulphate are pres-
ent in domestic wastewater but were not taken into account
for efficiency evaluation of treatment wetlands. First results
of laboratory-scale model investigations (Wiessner et al.,
2005) show highly efficient sulphate reduction in such systems
and indicate a correlation of sulphur transformation processes
with nitrogen and carbon removal. Usually domestic wastewa-
ter contains 20e500 mg l1 sulphate for theoretical maximum
sulphide generation of 7e167 mg l1. For example, sulphide
concentrations of even 0.5 mg l1 is known to be toxic for
microbial nitrification (AEsoy et al., 1998) and physiological
inhibition of several helophytes was shown for concentrations
of approximately 10e50 mg l1 (Armstrong et al., 1996;
Chambers et al., 1998; Fürtig et al., 1996). Furthermore, the
deposition of elemental sulphur in constructed wetlands treat-
ing domestic sewage as well as industrial wastewater
(Vymazal and Kröpfelová, 2005; Winter and Kickuth, 1989)
similar to paddy soil conditions may provide redox buffering
and a potential source for further redox processes (Wind and
Conrad, 1997; Zhou et al., 2002). Accordingly, particularly
under dynamic redox conditions of root-near environments
inside the rhizosphere sulphate reduction and oxidation of
sulphide, for example, and also disproportionation of several
sulphur compounds like elemental sulphur, sulphite, and
thiosulphate (Finster et al., 1998; Thamdrup et al., 1993) can
probably act simultaneously on small spatial scale.
Recently, the application of a specially designed labora-
tory-scale constructed wetland (Kappelmeyer et al., 2002) in
order to evaluate micro-gradient processes of root-near envi-
ronments of the rhizosphere was shown to be useful (Wießner
et al., 2005; Wiessner et al., 2005).
Using this technique, a long-term experiment was carried
out in this study using artificial domestic wastewater with
two different ratios of carbon-COD to sulphate loads. The
goal was to analyze the influence of this ratio of organic
carbon and sulphate on the extent of microbial sulphate reduc-
tion in the rhizosphere of constructed wetlands and about the
response of plants and microorganisms to sulphide generated
in various amounts during the process.
2. Material and methods
2.1. Experimental conditions
The model experiments were performed in a laboratory-scale wetland re-
actor under conditions of complete mixing of the pore water by a permanent
circulation flow. The internal flow conditions were comparable to conditions
of a continuously stirred tank reactor and, therefore, macro-scale gradients
of concentrations were equalized and the effects of gradient changes could
be determined. The design and the operation principles of the reactor (PFR-
planted fixed bed reactor) were previously described in detail (Kappelmeyer
et al., 2002; Wießner et al., 2005). Briefly, the rhizosphere of the reactor
was represented by a rooted gravel bed (particle size 2e4 mm) in a glass
reactor of 28 cm diameter and a height of 30 cm. The bed was planted with
five plants of Juncus effusus. The initial total number of shoots was 72 and
the plants growth developed a maximum total shoot number of 192 during
the experimental period. The pore water volume in the planted bed amounted
to 10 l and the hydraulic retention time was adjusted to 5 days. The reactor was
placed in a greenhouse and operated under defined environmental conditions
to simulate an average summer day in a moderate climate. The temperature
was set to 22  C from 6 am to 9 pm to simulate daytime and to 16  C at night.
One lamp (Master SON-PIA 400 W, Phillips, Belgium) was switched on
during daytime as an additional artificial light source whenever the natural
light fell below 60 klx.
A modified artificial wastewater (Wießner et al., 2005) with total carbon,
nitrogen, and sulphur concentrations according to a standard artificial domestic
wastewater (DIN-38412-T24, 1981) was used. The inflow concentrations were
(in mg l1): 107.1 C6H5COONa, 204.9 CH3COONa, 118 NH4Cl, 36.7
K2HPO4$3H2O, 7.0 NaCl, 3.4 MgCl2$6H2O, 4.0 CaCl2$2H2O, 222 Na2SO4
(dissolved in deionised water). The resulting concentrations of parameters
were (in mg l1): 326 COD, 37 ammonia-N, 50 sulphate-S, and 5 phos-
phate-P; the resulting molecular ratio of organic carbon (80% acetate þ 20%
benzoate) and sulphate was 2.8:1.
Before starting the experiment, the reactor was newly planted after highly
efficient operating for several months using similar artificial wastewater with-
out sulphate (Wießner et al., 2005). The experimental period started in March
2004 and continued until August 2005 for a total operation time of 552 days.
Thirty days after starting the experiment with sulphate-free inflow continuous
sulphate supply was started. After an operation time of 351 days carbon load
was doubled (COD ¼ 652 mg l1) resulting in a molecular ratio of organic
carbon to sulphate of 5.6:1.
Samples for sulphur compounds and COD analysis were taken once or
twice a week. Due to the reactor design, the circulation flow represents the
actual concentration of the pore water inside of the reactor and thus the pH,
redox potential, and the oxygen concentration were controlled continuously
in the circulation flow and the data were recorded online twice per hour.
Plant transpiration represents 98% of the total water loss (Wießner et al.,
2005) and was controlled by balancing the inflow and the outflow amounts
of water. The flow balances were also used to control and adjust the inflow
rate.
2.2. Analysis and calculations
COD was determined photometrically by using Test No. 314 over a range
of 15e150 mg l1 (Dr. Bruno Lange GmbH, Düsseldorf, Germany).
Concentrations of acetate, benzoate, ammonia, nitrite, and nitrate were an-
alysed by ion chromatography (DIONEX 100, columns AS4A-SC/AG4A-SC
and CS12A/CG12A; Idstein, Germany) using a UV-detector for nitrite and
nitrate at a wavelength of 215 nm and a conductivity detector for the other
ions. The self-regenerating suppressors ASRS-Ultra4-mm (for anions) and
CBES-I 4 mm (for ammonia) were used.
Sulphate was determined photometrically by using Spectroquant Test kit
over a range of 5e250 mg l1 (Merck, Germany). Sulphide was analysed pho-
tometrically using Test kit LCW 053 for 0.1e2 mg l1 (Dr. Bruno Lange
GmbH, Düsseldorf, Germany). After derivatisation using monobromobimane
sulphite and thiosulphate were determined by HPLC (Beckman, USA) using
fluorescence detector RF 551 (Shimadzu, Japan) and columns RP Li-Chros-
pher 60, RP Select B (250-4) according to Rethmeier et al. (1997). Elemental
sulphur was also determined according to Rethmeier et al. (1997), by extract-
ing samples with chloroform and subsequent detection by HPLC (Beckman,
USA) using a Li-Chrospher 100, RP 18 column (5 mm, Merck, Germany)
equipped with a UV-detector at 263 nm. Total sulphur was calculated by sum-
ming up all determined sulphur compounds.
Methane, carbon disulphide, and dimethylsulphide in the pore water were
directly analysed by membrane inlet mass spectrometry (MIMS) (Bohatka,
1997) using a QMG 422 quadrupole mass spectrometer (Pfeiffer Vacuum
 A. Wiessner et al. / Environmental Pollution 155 (2007) 125e131 127

GmbH, Germany) fitted with a silicon covered inlet probe (detection limit for

Outflow sulphur [mg Sl-1]

S-compounds: 0.1 mg l1). The quantification of methane concentrations was 60
based on calibrations using synthetic gases with 4e50% methane.
The elemental analysis (heavy metals, S and P) of the sediments was per-
formed by the energy dispersive X-ray fluorescence (EDXRF) spectrometer 40
XLAB 2000 (SPECTRO Instruments) running the software package X-LAB
Pro 2.2 after drying the sediment at 105  C for 24 h using oven MA4O (Sator-
20
ius, Germany). A mixed sample was used for analyses and means of triplicate
analyses were calculated. The method’s relative error is 2e3%. In order to
determine the content of total organics, the sediment was heated to 505  C 0
for 2 h using oven Hottherm 10 (Linn High Therm, Düsseldorf, Germany). Sulphate Sulphide
Determination of outflow amounts of total sulphur was based on area cal-
culation by Simpson-method (Press et al., 1994) using outflow concentration

Outflow sulphur [mg Sl-1]

12
of total sulphur.

3. Results 8

During the first part of the experiment characterized by

4
moderate COD supply (326 mg l1), a nearly complete COD
removal was observed. The dynamics of total sulphur and
the removal of COD are represented by the inflow and outflow 0
0 100 200 300 400 500
concentrations in Fig. 1. Outflow concentrations of total sul-
Time [d]
phur indicate varying sulphate removals. At the end of this ex-
perimental period (220e350 days), measured outflow Elemental sulphur Sulphite Thiosulphate
concentrations of total sulphur even exceeded the inflow
Fig. 2. Concentration of sulphate, sulphide, elemental sulphur, sulphite, and
values, which indicate the formation of S-deposits within the thiosulphate in the outflow.
reactor and/or increased concentrations by water losses of
plant transpiration. Nearly 100% of outflow sulphur (see
Fig. 2) was detected as sulphate, however, trace concentrations of outflow sulphate (see Figs. 1 and 2) was found to be corre-
(<1 mg l1) of sulphide, elemental sulphur, sulphite, and lated to the corresponding water losses by plant transpiration
thiosulphate were also observed during this moderate COD represented in Fig. 3B.
supply. The time course of sulphate removal calculated from A relatively stable pH of w7, healthy plant constitution
the outflow sulphate concentrations considering the water (increasing water loss by transpiration), and highly efficient
losses by the plants is represented in Fig. 3A. The enrichment biological nitrogen removal processes were observed (see
Fig. 3B, C and D) which means small residual ammonia
100 concentrations and no significant concentrations of nitrite
600 and nitrate indicating high efficiency of nitrification and
75
Removal (%)

denitrification.
COD [mgl-1]

400 After 351 days of operation total sulphur input amounted to

50 31 g S as sulphate and a total output of 24.1 g S mainly as sul-
200 phate and certain other S-species (see Figs. 1 and 2). The
25
resulting difference of 6.9 g S represents 22.2% of the total
0
S-input should be found to be deposited under the given highly
0
efficient micro gradient redox conditions.
Mean Inflow Outflow Removal
Doubling the carbon load from day 351 for the second part
of the experiment, COD removal continued on a high but
Total sulphur [mg Sl-1]

60 slightly lower level of approximately 85% (see Fig. 1). Sul-

phate reduction increased immediately to high levels, being
40
characteristic for this load (93% mean sulphate removal, see
Fig. 3A). With the highly efficient sulphate reduction, sulphur
deficits were found but over time, total sulphur of the outflow
20
increased to concentrations in the range of inflow-sulphate at
the end of the experiment (Fig. 1). Total sulphur of the outflow
0
0 100 200 300 400 500
mainly consists of reduced species such as sulphide and
Time [d]
elemental sulphur. At the end of the experiment about 70%
of total sulphur of the outflow was determined as sulphide,
Mean Inflow (Sulphate) Outflow
20% as elemental sulphur, and 10% as sulphate. Sulphite
Fig. 1. Inflow and outflow concentrations of total sulphur and COD and COD and thiosulphate were only minor compounds (mean concen-
removal; COD: chemical oxygen demand. trations of 0.16 mg l1 and 0.54 mg l1, respectively; Fig. 2).
128 A. Wiessner et al. / Environmental Pollution 155 (2007) 125e131

A 100 B
30
Sulphate Removal [%]

Water loss [ml h-1]

75

20
50

10
25

0 0
0 100 200 300 400 500 0 100 200 300 400 500
Time [d] Time [d]

C 8.0
D

Ammonia outflow [mg l-1]

30

7.5
20
pH

7.0
10

6.5 0
0 100 200 300 400 500 0 100 200 300 400 500
Time [d] Time [d]

Fig. 3. (A) sulphate removal (concentration by water losses accounted), (B) water loss by plant transpiration, (C) pH in the pore water and (D) ammonia concen-
tration in the outflow.

Calculation of the total sulphur for this period resulted in (representing nearly total ammonia removal) to the level of
16.9 g S of inflow, 11.2 g S of outflow, and 5.7 g S deficit, ac- the inflow concentration (40 mg l1) at the end of the experi-
cording to 33.8% of the total S-input being deposited inside ment. The correlation of the ammonia concentrations of the
the reactor. Due to the immediately raised pH-level from around outflow and the sulphide concentrations in the pore water is
7 to finally more than 7.5 mediated by microbial sulphate reduc- shown in Fig. 4 A, B. Outflow concentration of ammonia
tion (Raven and Scrimgeour, 1997) (Fig. 3A, C), losses of free started to increase at 0.5 mg l1 sulphide (see Fig. 4B). For
H2S from the surface of the gravel bed or by transport through sulphide concentrations up to a range of 5e10 mg l1,
the shoots should be of subordinated importance for the sulphur
balance of the system (Lens et al., 1998; Märkl, 1999). Sulphide A 40
uptake by the roots can be neglected (Fürtig et al., 1996). No sig-
40
nificant signals for carbon disulphide (CS2) and dimethylsul- 30 B
phide (S(CH3)2) were measured by mass spectrometry due to 20
the detection limits of 0.1 mg l1. In principle, this finding cor- 30 10
0
responds to measurements of emissions of various steps of the
Ammonia [mg l-1]

0.01 0.1 1 10 100

treatment processes of wastewater treatment plants (Devai and
DeLaune, 1999). 20
Plant transpiration decreased from 25 ml h1 (nearly 30%
of the inflow) before starting high C-load to finally
1.6 ml h1 (nearly 2% of the inflow) at the end of the experi-
ment (see Fig. 3B). Accordingly, the number of green healthy 10

shoots of nearly 200 at day 351 was reduced to a final remain-

der of 26 at the end of the second period of the experiment
(data not shown). 0
Similar to repression of plant activity (see Fig. 3B), the 0 10 20 30

removal of ammonia was found to be inhibited during the Sulphide [mg l-1]
second part of the experiment (Fig. 3D). The ammonia con- Fig. 4. Correlation of ammonia and sulphide outflow concentrations: (A)
centrations of the outflow increased from about 3 mg l1 non-logarithmical and (B) semi-logarithmical.
 A. Wiessner et al. / Environmental Pollution 155 (2007) 125e131 129

ammonia outflow concentrations increased to 12e22 mg l1 Table 1

(representing 45e70% ammonia removal). Further increase Average contents of elements in the sludge of the reactor in mg kg1 dry
weight
of sulphide concentration up to approximately 25 mg l1 did
not change ammonia removal (see Fig. 4A). Lead 6
Cadmium <2
Furthermore, only at the final stage of the experiment sig- Calcium 2001
nificant enrichment of methane could be measured reaching Chromium <10
a concentration of 7.4 mg l1 (34% saturation) in pore water Iron 1259
(Fig. 5). At the operation time around day 400, when plants Potassium 2739
damages became obvious by nearly halving the transpiration Copper 701
Manganese 134
rate (see Fig. 3 B) with only 80 residual green healthy shoots Nickel 122
from 200 initial ones (data not shown), methane was started to Phosphorus 11,011
be generated. Relating to further plant death, the concentration Total sulphur 21,100
of methane was found to be exponentially enriched towards Zinc 640
the end of the experiment simultaneously to highly efficient Elemental sulphur 0.45
sulphate reduction.
In order to gain more information about the fate of immobi-
lized sulphur, samples of the accumulated sludge in the gravel During S-cycling in the first part of the experiment, the spe-
bed were analysed at the end of the experiment. Total sulphur, cific root-near conditions of aerobic to anaerobic micro gradi-
elemental sulphur, heavy metals, and other element concen- ents caused by oxygen release from the roots have apparently
trations are shown in Table 1. The sludge consisted of approx- suppressed the intensity of microbial sulphate reduction
imately 2.1% total sulphur. However, small amounts of despite of a surplus of organic carbon as electron donor (see
elemental sulphur (<0.5 mg kg1) were found. The maximum Fig. 3A). The results indicate the importance of the plants
theoretical amount of sulphur of the sludge, which can be and of the well rooted environments inside treatment wetlands
bound by the heavy metals of the sludge (see Table 1) as heavy in order to guarantee oxidation/reduction equilibriums in
metal sulphides, is 1461 mg kg1 dry weight e this is only 7% S-cycling for undisturbed C and N removal and healthy plant
of the total sulphur amount of the sludge (see Table 1). A stocks (see Fig. 3B, D) mainly by repression of sulphide tox-
content of 95.1% of dry mass organic substances in sludge icity. The lack of significant amounts of elemental sulphur and
was estimated. thiosulphate in pore water (see Fig. 2) suggests that they are
either not formed or other highly efficient possible transforma-
tion reactions (reduction, oxidation, and disproportionation),
4. Discussion
or even other pathways finally result in the formation of a sig-
nificant S-depot being relatively stable under the given
Continuous stable microbial sulphate reduction activity even
conditions.
for a molecular ratio of carbon to sulphate of 1:1 was found for
Instantly starting highly efficient sulphate reduction in the
wastewater and soil treatment without plants (Barnes et al.,
second part of the experiment conditions immediately changed
1992; Seidel et al., 1995) whereas our findings for the actual mo-
(see Fig. 3A, C) and apparently caused sulphide toxicity to
lecular ratio of organic carbon to sulphate of 2.8:1 showed only
repress plant physiological activity. Therefore, advanced
partial sulphate reduction in the presence of the plants.
suppressions of internal gas ventilation and oxygen release
Sulphate reduction was always found for moderate carbon
by the roots of J. effusus can be considered for the measured
load but a sulphur balance of the in- and outflow concentra-
sulphide concentrations (see Fig. 2), as also found for other
tions indicates the immobilization of sulphur and the forma-
helophyte-species (Armstrong et al., 1996; Chambers et al.,
tion of a deposit.
1998; Fürtig et al., 1996). The resulting surplus of reduced
8 S-compounds by obviously insufficient reoxidation mainly
caused further enrichment of S2 finally to a high level of
35 mg l1 in pore water. Nevertheless, the simultaneous
6 enrichment of S0 finally to a maximum amount of 11 mg l1
Methane [mg l-1]

in the pore water suggests still active reoxidation of S2 to

S0 (Holmer and Storkholm, 2001).
4
Ammonia/sulphide correlation given in Fig. 4 strongly indi-
cates relatively high efficient possible nitrification in root-near
2 environments of the rhizosphere of J. effusus, despite of high
sulphide levels up to more than 20 mg l1. The toxicity limit
of 0.5 mg l1 sulphide (see Fig. 4B) for nitrification activity
0 in our rhizosphere biofilms was also found for biofilms in
300 350 400 450 500 550
sewers (AEsoy et al., 1998). For sulphide concentrations
Time [d]
higher than 25 mg l1 nitrification lasting continued to de-
Fig. 5. Concentration of methane in pore water. crease (see Figs. 2 and 3D). High variation of ammonia
130 A. Wiessner et al. / Environmental Pollution 155 (2007) 125e131
concentration in the outflow (16e32 mg l1) for sulphide con-
centration higher than 30 mg l1 (see Fig. 4A) suggests the re-
sistance of involved microorganisms to relatively high
sulphide concentrations. To explain this finding it can be pos-
tulated: (a) a biodiversity of nitrificants and selection of high
sulphide tolerant species and (b) a close association of nitrifi-
cants with the roots and their protection by oxygen released by
the roots (high level of redox potential, high level of reoxida-
tion of sulphide at root surfaces by oxygen).
Coexistence of methanogenesis and sulphidogenesis is well
known (Liesack et al., 2000). Nevertheless, for natural
wetlands it was shown that sulphate can substantially reduce
the methane formation (Gauci et al., 2004). Our results (see
Fig. 5), however, suggest repression of methanogenesis in
root-near environment by oxygen released by the plants. The
findings underline differences of the oxygen tolerance of
sulphate reducing and methane producing bacteria (Wind
and Conrad, 1997; Wind et al., 1999) and the importance of
different areas of the rhizosphere of helophytes for methane
emission from wetlands.
Due to the high content of organic substances in the sludge
(95.1% of dry mass), sulphur should be mainly immobilized
by organic substances of the sludge, such as dead plant matter
and microbial biomass. Presumably, S-deposition in root-near
environments of treatment wetlands with low pools of heavy
metals partly is similar to specific S-immobilization in peat
lands (Chapman, 2001; Gauci et al., 2004) and iron-poor
lake sediments (Holmer and Storkholm, 2001). Organic S of
approximately 64% C-bound S and 27% ester bound sulphate
was found to be dominant in peat (Chapman, 2001). Organic
sulphur compounds in lake sediments are less investigated
but may also include thiols, mercaptans, and thioethers
(Holmer and Storkholm, 2001).
5. Conclusions
 For carbon surplus conditions (carbon:sulphate of 2.8:1)
sulphate reduction happened but was repressed, in contrast
to unplanted filters mentioned in literature.
 No enrichment of reduced sulphur compounds was ob-
served in pore water but immobilization of sulphur in or-
ganic sludge could be determined.
 Only 7% of the total sulphur immobilized in the sludge
theoretically can be bound as heavy metal sulphide and
the formation of C-bounded organic S, ester sulphate,
thiols, mercaptans, and thioethers was postulated from
literature.
 High carbon load caused stable and efficient sulphate re-
duction, rising of pH, increasing enrichment of S2 and
S0 in pore water, and finally plant death and inhibition
of nitrification by sulphide toxicity.
 Inhibition of nitrification started at 0.5 mg l1 sulphide but
high residual nitrification efficiencies of up to 70% were
found even for high sulphide levels of 6e25 mg l1.
 No significant enrichments of sulphite, thiosulphate,
carbon disulphide, and dimethylsulphide were observed
during the experiment.
 Methane production was correlated to plant death, sug-
gesting repression of methanogenesis by oxygen released
by healthy plant roots. Maximum concentration of
7.4 mg l1 (34% saturation) was measured in pore water.
 Stability and reoxidation of immobilized S and correlation
of S-dynamics and methanogenesis in root-near environ-
ments are objects of further investigation.
Acknowledgements
The work was funded by a grant of the UFZ. The authors
like to thank Kerstin Puschendorf, Ines Mäusezahl, Reinhard
Schumann, Nora Puschmann, Claudia Pietsch, and Doris
Sonntag for technical assistance.
References
AEsoy, A., Odegaard, H., Bentzen, G., 1998. The effect of sulphide and
organic matter on the nitrification activity in a biofilm process. Water
Science and Technology 37 (1), 115e122.
Armstrong, J., AfreenZobayed, F., Armstrong, W., 1996. Phragmites die-back:
sulphide- and acetic acid-induced bud and root death, lignification, and
blockages within aeration and vascular systems. New Phytologist 134
(4), 601e614.
Barnes, L.F., Janssen, F.J., Scheeren, P.H.J., Versteegh, J.H., Koch, R.O., 1992.
Simultaneous microbial removal of sulphate and heavy metals from waste
water. Transactions of the Institution of Mining and Metallurgy C: Mineral
Processing and Extractive Metallurgy 101, C183eC189.
Bezbaruah, A.N., Zhang, T.C., 2004. pH, redox, and oxygen microprofiles in
rhizosphere of bulrush (Scirpus validus) in a constructed wetland treating
municipal wastewater. Biotechnology and Bioengineering 88 (1), 60e70.
Bohatka, S., 1997. Process monitoring in fermentors and living plants by
membrane inlet mass spectrometry. Rapid Communications in Mass
Spectrometry 11 (6), 656e661.
Chambers, R.M., Mozdzer, T.J., Ambrose, J.C., 1998. Effects of salinity and
sulfide on the distribution of Phragmites australis and Spartina alterniflora
in a tidal saltmarsh. Aquatic Botany 62 (3), 161e169.
Chapman, S.J., 2001. Sulphur forms in open and afforested areas of two
Scottish peatlands. Water, Air, and Soil Pollution 128, 23e39.
Choi, J.H., Park, S.S., Jaffè, P.R., 2006. The effect of emergent macrophytes
on the dynamics of sulfur species and trace metals in wetland sediments.
Environmental Pollution 140, 286e293.
Devai, I., DeLaune, R.D., 1999. Emission of reduced malodorous sulphur
gases from wastewater treatment plants. Water Environment Research 71
(2), 203e208.
DIN-38412-T24, 1981. German Standard Methods for the Analysis of Water,
Waste Water and Sludge e Bio-Assays (Group l): Determination of Biode-
gradability by Use of Special Methods of Analysis, L 24. Beuth Verlag
GmbH, Hennef, Germany.
Finster, K., Liesack, W., Thamdrup, B., 1998. Elemental sulphur and thiosul-
fate disproportionation by Desulfocapsa sulfoexigens sp. nov., a new anaer-
obic bacterium isolated from marine surface sediment. Applied and
Environmental Microbiology 64 (1), 119e125.
Fürtig, K., Rüegsegger, A., Brunold, C., Brändle, R., 1996. Sulphide utilization
and injuries in hypoxic roots and rhizomes of common reed (Phragmites
australis). Folia Geobotanica and Phytotaxonomica 31, 143e151.
Gauci, V., Fowler, D., Chapman, S.J., Dise, N.B., 2004. Sulfate deposition and
temperature controls on methane emission and sulfur forms in peat.
Biogeochemistry 71 (2), 141e162.
Holmer, M., Storkholm, P., 2001. Sulphate reduction and sulphur cycling in
lake sediments: a review. Freshwater Biology 46 (4), 431e451.
Kalin, M., 2004. Passive mine water treatment: the correct approach? Ecolog-
ical Engineering 22, 299e304.
 A. Wiessner et al. / Environmental Pollution 155 (2007) 125e131
Kappelmeyer, U., Wießner, A., Kuschk, P., Kästner, M., 2002. Operation of
a universal test unit for planted soil filters e planted fixed bed reactor.
Engineering in Life Sciences 2, 311e315.
Lamers, L.P.M., Falla, S.J., Samborska, E.M., van Dulken, L.A.R.,
van Hengstum, G., Roelofs, J.G.M., 2002. Factors controlling the extent
of eutrophication and toxicity in sulfate-polluted freshwater wetlands.
Limnology and Oceanography 47 (2), 585e593.
Lens, P.N.L., Visser, A., Janssen, A.J.H., Hulshoff Pol, L.W., Lettinga, G.,
1998. Biotechnological treatment of sulfate-rich wastewaters. Critical
Reviews in Environmental Science and Technology 28 (1), 41e88.
Liesack, W., Schnell, S., Revsbech, N.P., 2000. Microbiology of flooded rice
paddies. FEMS Microbiology Reviews 24, 625e645.
Märkl, H., 1999. Modeling of biogas reactors. In: Rehm, H.-J., Reed, G.,
Pühler, A., Stadler, P. (Eds.), Biotechnology, vol. 11a. Wiley-VCH,
Weinheim, Germany, pp. 527e560.
Morrison, M., Aplin, A.C., 2003. CeSeFe geochemistry of a wetland
constructed to treat acidic mine waters. Land Contamination and Reclama-
tion 11 (2), 239e244.
O’Sullivan, A.D., McCabe, O.M., Murray, D.A., Otte, M.L., 1999. Wetlands
for rehabilitation of metal mine wastes. Biology and Environment:
Proceedings of the Royal Irish Academy 99B (1), 11e17.
O’Sullivan, A.D., Moran, B.M., Otte, M.L., 2004. Accumulation and fate of con-
taminants (Zn, Pb, Fe and S) in substrates of wetlands constructed for treat-
ing mine wastewater. Water, Air, and Soil Pollution 157 (1e4), 345e364.
Press, W.H., Teukolsky, S.A., Vetterling, W.T., Flannery, B.P., 1994. Numeri-
cal Recipes in Fortran. Cambridge University Press, New York, USA.
Raven, J.A., Scrimgeour, C.M., 1997. The influence of anoxia on plants of sa-
line habitats with special reference to the sulphur cycle. Annals of Botany
79, 79e86.
Rethmeier, J., Rabenstein, A., Langer, M., Fischer, U., 1997. Detection of
traces of oxidized and reduced sulfur compounds in small samples by
combination of different high-performance liquid chromatography
methods. Journal of Chromatography A 760 (2), 295e302.
131
Seidel, H., Ondruschka, J., Kuschk, P., Stottmeister, U., 1995. Influence of the
sulphur content in sediments on the mobilization of heavy metals by
bacterial leaching. Vom Wasser 84, 419e430.
Thamdrup, B., Finster, K., Hansen, J.W., Bak, F., 1993. Bacterial dispropor-
tionation of elemental sulfur coupled to chemical-reduction of iron or
manganese. Applied and Environmental Microbiology 59 (1), 101e108.
Vymazal, J., Kröpfelová, L., 2005. Sulphur deposits at the outflow from
horizontal sub-surface flow constructed wetlands e what does it indicate?
In: International Symposium on Wetland Pollutant Dynamics and Control,
Sept. 4e8, 2005, Ghent, Belgium, Book of Abstracts, pp. 200e201.
Webb, J.S., McGinness, S., Lappin-Scott, H.M., 1998. Metal removal by sul-
phate-reducing bacteria from natural and constructed wetlands. Journal of
Applied Microbiology 84, 240e248.
Wei, S., Zhou, Q., Zhang, K., Liang, J., 2003. Roles of rhizosphere in remedi-
ation of contaminated soils and its mechanisms. Chinese Journal of
Applied Ecology 14 (1), 143e147.
Wiessner, A., Kappelmeyer, U., Kuschk, P., Kästner, M., 2005. Sulphate reduc-
tion and the removal of carbon and ammonia in a laboratory-scale
constructed wetland. Water Research 39, 4643e4650.
Wießner, A., Kappelmeyer, U., Kuschk, P., Kästner, M., 2005. Influence of the
redox condition dynamics on the removal efficiency of a laboratory-scale
constructed wetland. Water Research 39, 248e256.
Wind, T., Conrad, R., 1997. Localization of sulfate reduction in planted and
unplanted rice field soil. Biogeochemistry 37, 253e278.
Wind, T., Stubner, S., Conrad, R., 1999. Sulfate-reducing bacteria in rice field
soil and on rice roots. Systematic and Applied Microbiology 22 (2),
269e279.
Winter, M., Kickuth, R., 1989. Elimination of sulphur compounds from waste-
water by the root zone process e II. Mode of formation of sulphur
deposits. Water Research 23 (5), 547e560.
Zhou, W., Wan, M., He, P., Li, S.T., Lin, B., 2002. Oxidation of elemental
sulfur in paddy soils as influenced by flooded condition and plant growth
in pot experiment. Biology and Fertility of Soils 36 (5), 384e389.