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Abstract
. Technological aspects;
. The effect of root growth on the soil matrix;
. Gas transport in helophytes and the release of oxygen into the rhizosphere;
. The uptake of inorganic compounds by plants;
. The uptake of organic pollutants by plants and their metabolism;
. The release of carbon compounds by plants;
. Factors affecting the elimination of pathogenic germs.
0734-9750/$ - see front matter D 2003 Elsevier Inc. All rights reserved.
doi:10.1016/j.biotechadv.2003.08.010
94 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
1. Introduction
Fig. 1. Possible interactions in the root zone of wetlands for wastewater treatment.
U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117 95
Table 1
Selection of plant species used in constructed wetlands
Scientific name English name
Phragmites australis (Cav.) Trin. ex Steud. common reed
Juncus spp. rushes
Scirpus spp. bulrushes
Typha angustifolia L. narrow-leaved cattail
Typha latifolia L. broad-leaved cattail
Iris pseudacorus L. yellow flag
Acorus calamus L. sweet flag
Glyceria maxima (Hartm.) Holmb. reed grass
Carex spp. sedges
where the only issues of concern were the inlet and outlet loads. This is almost solely
accounted for by the lack of suitable testing systems and study methods. However, small-
scale process modelling experiments are currently being developed (Kappelmeyer et al.,
2002).
According to practical experience and corresponding experiments, species of helo-
phytes (marsh plants) work best of all in seminatural wastewater treatment systems. This is
because helophytes possess specific characteristics of growth physiology that guarantee
their survival even under extreme rhizosphere conditions. The extreme conditions in the
rhizosphere in wetlands used to treat wastewater can be summed up as follow:
Highly reduced milieu (Eh up to < 200 mV, especially in horizontal subsurface flow
systems) prompting the formation of H2S and CH4;
Acidic or alkaline pH values in certain wastewaters;
Toxic wastewater components such as phenols, tensides, biocides, heavy metals, etc.;
Salinity.
Although all the plant species listed in Table 1 are suitable, reeds along with types of
rushes and cattails are the ones most frequently used. Recently, the suitability of fast-
growing trees such as willows has also been examined (Greenway and Bolton, 1996).
In order to learn more about the complexities of what happens when organic pollutants
are degraded in the root zone, we need to know more about the plants’ physiological
peculiarities and the microorganisms active in their rhizospheres.
2. Technological aspects
The knowledge accumulated over time about ways in which contaminants can be
removed by the simple natural combination of water, plants and soils has led to the
deliberate application of such systems in nature and ultimately to the creation of artificial
systems with various states of naturalness. Wissing (1995) divides the systems into the
following three main groups (see also Fig. 2):
Aquaculture systems. Installations without active soil filters, such as ponds and ditches
with intensive growth of submerged aquatic and/or free-floating plants (Xu et al., 1992);
96 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
Fig. 2. Pond/wetland systems for wastewater treatment (A, pond with free-floating plants; B, horizontal surface
flow wetland or pond with emergent water plants; C, horizontal subsurface flow wetland; D, vertical flow
wetland).
Hydrobotanical systems. Installations with a few active soil filters where removal is
mainly effected by aquatic plants, helophytes and microorganisms—ponds and ditches
with intensive growth of mainly helophytes;
Soil systems (see Fig. 2 for more details).
Horizontal surface flow systems (with the wastewater level above the soil surface);
Horizontal subsurface flow systems (with the wastewater level below the soil surface);
Vertical flow systems with upstream or downstream characteristics and continuous or
intermittent loading.
has prompted intensive technological research and development in recent years. As far as
mainly domestic wastewater treatment is concerned, investigations have focused on the
following aspects:
soil hydraulics (the influence of soil matter on hydraulic permeability), (Sanford et al.,
1995);
flow characteristics (horizontal or vertical flow, continuous or discontinuous water
load) (Netter, 1992; Stairs and Moore, 1994; Chazarenc et al., 2002);
external oxygen supply (by waterfalls, overflows and aeration installations, often in
connection with defined soil formations and discontinuous water loading or external air
input) (Green et al., 1998);
minimizing the area needed and maximizing the input load of contaminants; varying
the hydraulic retention time (Platzer and Netter, 1994; Platzer, 1999);
construction configuration, coupling different systems (Vymazal and Masa, 2002);
the plants used (empirical exploitation of different plants, mono vs. mixed culture;
influence of root growth) (Breen and Chick, 1995).
One important aspect of the complex processes taking place in the rhizosphere is the
interaction between roots/rhizomes and the soil matrix. The soil is the main supporting
material for plant growth and microbial films. Moreover, the soil matrix has a decisive
influence on the hydraulic processes.
Both chemical soil composition and physical parameters such as grain-size distribu-
tions, interstitial pore spaces, effective grain sizes, degrees of irregularity and the
coefficient of permeability are all important factors influencing the biotreatment system.
These physical parameters indicate certain hydraulic states of the soil and considerably
influence the flow of wastewater in constructed wetlands—and ultimately the removal of
contaminants. Root growth affects the physical (hydraulic) quality of soils (Kickuth, 1984;
98 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
Cooper and Boon, 1987; Wissing, 1995). On the one hand, roots and microbial biomass
clog up soil pores, but on the other hand, root growth and the microbial degradation of
dead roots cause the formation of new secondary soil pores.
As far as constructed wetlands are concerned, it seems that the main parameter
influencing the soil hydraulics is the grain-size distribution. Experience in Germany and
long-term studies of the hydraulics of constructed wetlands with different soil parameters
indicate that a mixture of sand and gravel produces the best results in terms of both
hydraulic conditions and the removal of contaminants (Wissing, 1995; Börner, 1990;
Netter, 1990).
For constructed wetlands with vertical flow, a relatively small range of effective grain
size d10 (the grain size below which 10 wt.% of the soil consists of, and which is the
leading characteristic of soils) from 0.06 to 0.1 mm was evaluated, while that for
constructed wetlands with horizontal flow was found to be higher at 0.1 mm (because
of the higher susceptibility to obstruction) (Wissing, 1995). The grain-size distribution of
>0.06 mm (up to 10 mm) with different distribution characteristics apparently enables
effective coefficients of permeability in the range of >10 5 m/s (Wissing, 1995; Bahlo and
Wach, 1993), enough immobilization surface area for biofilm growth, positive impacts of
root growth on the hydraulic conductivity of the soil, and hence, all in all the good removal
of contaminants.
Investigations were conducted on a whole series of constructed wetlands which were
exclusively installed using soil material with Kf < 10 8 m/s in order to maximize the area
for biofilm growth and the adsorption of wastewater chemicals (Morell, 1990; Börnert,
1990; Netter, 1990; Kretzschmar, 1990). The systems suffered hydraulic problems mainly
because of short-circuit flow on the wetland surface, and the predicted improvements in
the hydraulic conditions by root growth in the course of time were not observed. Despite
small increases in the Kf values (up to about 10 7 m/s), the hydraulic conditions did not
change sufficiently. The main root growth of Phragmites australis was only recorded in
the top soil zone down to a depth of 20 –30 cm (Börnert, 1990).
In intermittently charged vertical filters, oxygen mainly enters the soil filter by virtue of
the suction effect of the water as it flows downwards. By contrast, in subsurface horizontal
flow systems oxygen is chiefly input by the marsh plants (helophytes).
How higher plants react to a lack of oxygen in the rhizosphere varies. Whereas typical
land plants which are adapted to dry locations cannot survive for long under such
conditions, plants which are adapted to waterlogged areas such as marshes, moors,
swamps and riverbanks have the anatomical and physiological attributes necessary for
their long-term survival (Vartapetian and Jackson, 1997).
The degree of adaptation is specific to individual species. Their survivability varies
over an extremely broad tolerance band from a few hours to several months, as
demonstrated by experiments in anaerobic incubators (Crawford and Braendle, 1996).
The fact that plants adapted to anoxic rhizosphere conditions can survive is because of
their ability to supply their root system with oxygen from the atmosphere. Gas transport
U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117 99
from the sections of the plant above the ground through the rhizome into the fine roots is
effected by specific areas of tissue formed in the plant known as the aerenchyma.
Depending on the degree of adaptation, these gas chambers can account for as much as
60% of the total tissue volume (Grosse and Schröder, 1986). The gas chambers in the
rhizome area are protected by node-like segmentation and diaphragms which are gas
permeable but nevertheless provide a secure barrier which prevents liquids from pene-
trating (Soukup et al., 2000). The various possibilities involved in the genesis of
aerenchyma structures by cell lysis or cell formation as well as their anatomical
peculiarities have been (and remain) the subject of thorough anatomical and physiological
studies (Jackson and Armstrong, 1999; Drew, 1997; Allen, 1997; Armstrong et al., 1994).
Interest is focused on issues such as how the interaction of changing environmental
conditions in the rhizosphere and biochemical processes causes anatomical changes in the
plant (Jackson and Armstrong, 1999).
The flow of gas through the plants is driven by diffusion processes and/or intensive
convective flows inducing high and low pressure (Allen, 1997; Armstrong et al., 1991;
Jackson and Armstrong, 1999; Grosse and Frick, 1999; Grosse, 1989). The types and
combination of the mechanisms involved are specific to each plant. For example, very
intensive convective gas transport has been observed in Typha latifolia (cattail) and P.
australis (reed) (Bendix et al., 1994; Armstrong and Armstrong, 1991). This convection is
caused by the formation of low pressure in oxygen-consuming sections of the plant and the
formation of higher pressure in the plant’s leaves (Allen, 1997). The formation of low
pressure is mainly based on the different solubilities of the oxygen used for restoration and
the carbon dioxide formed in this process. The formation of higher pressure in the leaves
causes air to flow throughout the entire body of the plant, with transport rates of up to 10
ml air per minute being measured (Grosse and Schröder, 1986; Schröder, 1986). One of
the main processes causing higher pressure is thermoosmosis (Grosse and Schröder, 1986;
Allen, 1997).
Owing to the differences in temperature between the cold phylloplane and the warmer
leaf interior, thermoosmosis causes air molecules to enter the young leaves through pores
(which are smaller than those in older leaves). The warmer interior of the leaf causes the
gas to expand owing to Brownian movement, limiting the possibility of returning through
the leaf pores. The overpressure building up inside the leaves is compensated for in the gas
transport tissue (the aerenchyma) inside the plant. As a result, the gas molecules are
transported through the plant right down to the deepest roots. The pressure compensation
of the plant system is finally achieved by gas being released through the roots and through
older leaves with larger pores.
The processes involved in the pressure-induced flow of gas in plants have been studied
since the mid-19th century (Grosse et al., 1996). This interest in the gas balance of higher
plants has been increasingly revived since the 1980s, not least in connection with the
growing interest in the biotechnological usage of plants adapted to flooded conditions, for
example, in order to clean wastewater (Grosse et al., 1996).
The introduction of atmospheric air into the plant’s interior means that under anoxic
conditions a sufficient amount of oxygen is available in the rhizome and root zones,
which can be used for respiration. However, the oxygen transported in the airflow is also
vital to the plant’s survival in another respect. Oxygen is released into the rhizosphere
100 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
and parts of the root system, mainly around the root tips and on young laterals
(Armstrong et al., 1990; Flessa, 1991). The release of oxygen causes the formation of
an oxidative protective film directly on the root surface. This film protects the sensitive
root areas from being damaged by toxic components in the anoxic, usually extremely
reduced rhizosphere (Armstrong et al., 1994; Vartapetian and Jackson, 1997). This
protective film has a thickness of between 1 and 4 mm depending on the way in which
incoming oxygen-consuming wastewater flows against the roots, and it contains redox
gradients ranging from about 250 mV as frequently measured in reduced rhizospheres
to about + 500 mV directly on the root surface (Flessa, 1991). Oxygen is continuously
released from the internal root zones, counterbalancing chemical and biological oxygen
consumption.
This constant release of oxygen in the rhizosphere is of particular interest in connection
with the exploitation of the rhizosphere to treat wastewater. The oxygen flow rates which
have been measured, e.g., 126 Amol O2/h g root dry mass for Juncus ingens (giant rush)
(Sorrell and Armstrong, 1994) and 120 –200 Amol O2/h g root dry mass for T. latifolia
(cattail) (Jespersen et al., 1998) are of biotechnological relevance. Model calculations for
P. australis (reed) resulted in area-specific oxygen input rates of 5– 12 g O2/m2 patch area
per day (Armstrong et al., 1990). If the oxidation potential of certain helophytes is to be
put to optimal biotechnological use, this process must be quantified taking into account
the various factors of influence. These factors include rhizosphere-specific parameters
such as the redox state, pH, oxygen concentration, chemical characteristics and
temperature, plant-specific parameters such as mass and the species and stage of
development of plants, as well as phylloplane-specific factors such as temperature and
light intensity.
Studies have revealed that the redox state of the rhizosphere has a significant effect on
the intensity of oxygen release through the roots of various helophytes (Sorrell and
Armstrong, 1994; Sorrell, 1999; Kludze and Delaune, 1996; Wießner et al., 2002a). For
example, it was found that the release intensities clearly depend on the redox state of the
medium in the hydroponic vessel for T. latifolia and Juncus effusus plantlets under reduced
conditions, as shown in Fig. 3.
The oxygen release rates were highest at 250 mV < Eh < 150 mV. For extremely
reduced rhizospheric conditions (Eh < 250 mV) and also moderately reduced rhizo-
spheric conditions (Eh> 150 mV), the release intensities were found to be lower. The
potential of T. latifolia to release oxygen was much higher than that of J. effusus. Under
initially oxygen-free conditions but already in the positive redox range, all the plants of the
two species continued to release oxygen up to highly oxidized states.
Another interesting result of laboratory experiments is the correlation between root and
shoot size and oxygen release into the rhizosphere, as shown in Figs. 4 and 5. The relative
independence of the oxygen release rate from the root size and the significant influence of
the shoot size are evident.
For an explanation of this finding, it is well known that only small parts of the whole
root system (root tips and laterals) are permeable enough to transfer gases.
Otherwise, aspects of the aboveground biomass such as leaf areas and stomatal
conductance appear to be very important plant-specific parameters, including the plants’
capacities to release oxygen into their rhizosphere. The correlation between oxygen release
U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117 101
Fig. 3. Maximum oxygen release rates of individual experiments with several plantlets of T. latifolia, P. australis
and J. effusus depending on the corresponding redox potential (adapted from Wießner et al., 2002a, with
permission).
rates and shoot size can be described mathematically by nonlinear equations for both
species (see Fig. 6) (Wießner et al., 2002a).
Illumination was found to influence the intensity of oxygen release for T. latifolia
decisively but less so for J. effusus (Fig. 7). Obvious processes of aboveground gas
exchange and/or gas transport inside the plants (photosynthesis, thermoosmosis, diffusion,
Fig. 4. Oxygen release rates of J. effusus correlated with root and aboveground biomasses. ORR: oxygen release
rate; ADW: aboveground dry weight; RDW: root dry weight. ORR values are the mean of three experiments with
each plant (range of relative deviations 4 – 17%) (adapted from Wießner et al., 2002b, with permission).
102 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
Fig. 5. Oxygen release rates of J. effusus correlated with root and aboveground biomasses. ORR: oxygen release
rate; ADW: aboveground dry weight; RDW: root dry weight. ORR values are the mean of three experiments with
each plant (range of relative deviations 4 – 17%) (adapted from Wießner et al., 2002b, with permission).
Fig. 6. Oxygen release rates of T. latifolia and J. effusus correlated to aboveground dry weight (adapted from
Wießner et al., 2002b, with permission).
U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117 103
Fig. 7. Oxygen release rates of T. latifolia and J. effusus correlated to illumination (adapted from Wießner et al.,
2002b, with permission).
rice plant (Banker et al., 1995). Thomas et al. (1996) summarized and cited other papers in
which helophytes are responsible for 50 –90% of the total methane flux from wetlands.
Tanner et al. (1997) estimated methane emissions from constructed wetlands used to treat
agricultural wastewater to account for around 2– 4% of wastewater carbon loads in
vegetated wetlands and 7 – 8% of loads in unvegetated systems.
Other pollutants (phytovolatilization) such as trichloroethylene and 2,6-dimethylphenol
were only emitted in trace amounts and are not significant for the technological treatment
process (Baeder-Bederski-Anteda, 2002).
The main mechanisms of nutrient removal from wastewater in constructed wetlands are
microbial processes such as nitrification and denitrification as well as physicochemical
processes such as the fixation of phosphate by iron and aluminum in the soil filter.
Moreover, plants are able to tolerate high concentrations of nutrients and heavy metals,
and in some cases even to accumulate them in their tissues.
The amount of phosphorous which can be taken up in the surface biomass of
Schoenoplectus lacustris (Sch. lacustris) is about 6.7 g m2 a 1 (Seidel, 1966). The mean
phosphorous content in the dry biomass of a large number (41) of helophytes was found
by McJannet et al. (1995) to be around 0.15 –1.05%. Consequently, less than 5% of the
phosphorus load in municipal wastewater is taken up by the plants. Seen from this angle,
the effect of harvesting the plant biomass is insignificant (Kim and Geary, 2001).
The uptake of nitrogen into the plant biomass is also of minor importance from a technical
viewpoint since harvesting the aboveground biomass would remove only 5– 10% of the
nitrogen (Thable, 1984). Tanner (1996) estimated the nitrogen concentrations in helophytes
in the aboveground biomass to be between 15 and 32 mg N g 1 dry mass. Owing to these
relatively low levels of nutrients, plant biomass is usually not harvested in Europe.
104 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
One of the pioneers of using helophytes to treat wastewater was Seidel (1968). She
was the first person to study the removal of various phenols by helophytes in
hydroponic vessels as well as the tolerance of plants to phenols. Because these studies
were carried out as batch experiments under nonsterile conditions, the uptake is likely to
have been caused by both microorganisms and the plants themselves. In addition, no
constant test concentrations were established during her investigation of tolerance to
contaminants.
Infusion experiments with sterile plant tissue of Scirpus lacustris (Sc. lacustris) showed
an uptake of 0.08 mg phenol g 1 fresh mass day 1 (Kickuth, 1970). The main metabolite
identified was picolinic acid. It was therefore surmised that phenol degradation chiefly
takes place via catechol and further meta-ring cleavage. In the case of Lemna gibba,
U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117 105
Transformation;
Conjugation;
Compartmentation.
Cytochrome P450;
Glutathione transferase;
Carboxylesterase;
O- and N-glucosyl transferase;
O- and N-malonyl transferase;
The current knowledge about the input of carbon from plants into their rhizosphere
comes mainly from agricultural research.
The entire process of carbon input is known as rhizodeposition. Rhizodeposition
products (exsudates, mucigels, dead cell material, etc.) cause various biological processes
to take place in the rhizosphere. The quantity of organic carbon compounds released has
been estimated at 10– 40% of the net photosynthetic production of agricultural crops
106 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
(Helal and Sauerbeck, 1989). The chemical composition of the exudates is very diverse.
Compounds which occur in plant tissues are usually also released through the roots. For
example, among the substances which have been identified in root exudates are sugars and
vitamins such as thiamine, riboflavin and pyridoxine etc., organic acids such as malate,
citrate, amino acids, benzoic acids, and phenol and other organic compounds (Miersch et
al., 1989). The range of substances varies from one species or even subspecies to the next.
It is assumed that the rhizodeposition products perform the following functions in the
rhizosphere:
Mobilizing nutrients. Nutrient limitation can cause organic acids or other compounds to
be excreted. This may for example increase the solubility of iron and phosphate, thus
improving the plant’s nutrient supply (Hoffland et al., 1992).
Allelopathic effects. Some species of plants excrete special compounds into the
rhizosphere which impede the growth of other plant species (Miersch et al., 1989).
This effect has been examined in detail for a number of agricultural crops. The
literature does not yet contain any clear indications of allelopathy among helophytes
(Gopal and Goel, 1993).
Rhizosphere effects. Organic compounds such as sugars and amino acids can be used
by microorganisms as substrates, and excreted vitamins stimulate microbial growth.
Helal and Sauerbeck (1989) report that the majority of organic compounds excreted by
maize (80%) are mineralized by the microorganisms in the rhizosphere to form CO2,
increasing the microbial biomass in the rhizosphere. Furthermore, it has been shown
that organic compounds released by plants and plant residues influence the microbial
degradation of xenobiotics (Horswell et al., 1997; Donnelly et al., 1994; Moormann et
al., 2002). This issue is examined in more detail in Section 9.
Current knowledge of the composition of root exudates of helophytes is very limited, and
so far almost nothing is known about adult plants in this regard. Kaitzis (1970) investigated
rhizome extracts of Sc. lacustris and found various benzene derivatives including hydroxyl,
methoxyl, aldehyde and carboxyl groups. These extracted compounds displayed bactericidal
effects, indicating that they are responsible for the ‘negative’ rhizosphere effect (for more
details, see Section 10).
Owing to the relatively low amount of carbon released by plants in comparison to the
water flow, it can be assumed that rhizodeposition is only significant in constructed
wetlands if the carbon load in the wastewater is extremely low, as is for instance the case
with mine drainage. Rhizodeposition products can be used for bacterial dissimilatory
sulfate reduction. The H2S arising combines with heavy metal ions to form poorly soluble
sulfides in the anaerobic areas of the rhizosphere.
8. Transpiration
5NHþ
4 þ 3NO3 ! 4N2 þ 9H2 O þ 2H
þ
first specified by Van de Graaf et al. (1990) plays a significant part in this system.
In constructed wetlands, especially subsurface horizontal flow systems, very little
attention has been paid to the sulfur metabolism. In the case of an industrial wastewater
loaded with SO42 and S2O32 (area-specific load of 1.1 g S/m2 day), Winter (1985)
showed that constructed wetlands can act as an important sink for sulfur. Two percent of
the load was retained in the soil: 31% as S0, 25% as organic S (mainly in humic matter),
15% as sulfate and 11% as sulfide. Both microbial and abiotic processes are responsible
for these transformation processes.
108 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
Heavy metals are usually removed from industrial wastewater and mine drainage in
constructed wetlands by a variety of methods including:
In the aqueous phase of surface flow wetlands to treat mine drainage, Fe(II) is oxidized
to Fe(III) by abiotic and microbial oxidation; other elements such as arsenic also
precipitated. Other heavy metals are immobilized in the mainly anoxic soil by microbial
dissimilatory sulfate reduction and the H2S formed.
As far as highly chlorinated organic compounds are concerned, it is always much
easier for the corresponding carbon atom to be attacked by nucleophilic rather than by
oxidative reactions. Therefore, highly chlorinated hydrocarbons have a much higher
chance of being dehalogenated than less chlorinated compounds. Adrian et al. (1998)
demonstrated the microbial reduction of trichlorobenzene to form monochlorobenzene
via dichlorobenzene.
The only realistic possibility of biologically degrading hexachlorobenzene perchlo-
roethylene or highly chlorinated biphenyls is reductive dehalogenation (Wischnak and
Müller, 2000). The low-chlorinated products can then undergo further biological
degradation under aerobic conditions. Owing to the different redox states, existing as
It was back in the 1970s that Seidel (1971, 1972, 1973a,b) first drew attention to the
bactericidal effect of higher plants on pathogenic germs. She put this aspect to practical
use in the first constructed wetlands built in Germany. Her laboratory experiments revealed
that the effect varied depending on the species of plant used. For example, 10 plant species
managed to remove 99% of Escherichia coli in pot experiments within 48 h, while 13
species removed 85% and another 31 species only eliminated 15%. Among the helo-
phytes, Mentha aquatica, Alisma plantago and J. effusus proved to be especially efficient
(Seidel, 1971). Seidel’s findings were largely confirmed by Burger and Weise (1984) in
pot experiments using 1.2 l of sand and 5 l of nutrient solution. In pots containing Glyceria
maxima, S. lacustris, A. plantago-aquatica and M. aquatica, the number of bacteria
(colony-forming units) was reduced by 90% after a contact time of 7 –11 h and by 99%
after 16 –19 h. The effectiveness was highest in the first few days of the investigation
period. Compared to the control experiments, the reduction time was between a third and a
half shorter.
Unfortunately, the findings of the two groups (Seidel vs. Burger and Weise) cannot be
directly compared since the descriptions of the experimental method (the way in which the
experiments were carried out, the state of the plants, etc.) are too incomplete.
Vincent et al. (1994) studied the bactericidal effect of the in vitro (aseptically)
cultivated helophytes M. aquatica, P. australis and Sc. lacustris on E. coli. In the test
system, all three plant species inhibited the growth of E. coli, with the strongest effect
being exhibited by Sc. lacustris. Only in the case of M. aquatica did plant-free nutrient
110 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
solutions from which the plants had been removed after 14 days’ cultivation show a slight
antibacterial effect. The authors concluded that the bactericidal effect is an active process
that requires the direct presence of plants.
Despite these findings, it is difficult to explain the fact that the root exudates both
stimulate and inhibit bacterial growth. Therefore, other mechanisms and indirect effects by
plants (adsorption, aggregation and filtration) as well as the effect of protozoa have been
discussed (Kadlec and Knight, 1996).
The exact role of biolytic processes (such as the action of protozoa, bdellovibrios and
bacteriophages) in germ reduction in constructed wetlands is still largely unknown.
Gradl et al. (1994) carried out small-scale experiments into the removal of fecal coli and
total coli from the discharge of a mechanical – biological clarification plant with a
vertically charged soil filter system (2.5 m/day). The filter was planted with Acorus
calamus and P. australis. The filter planted with A. calamus achieved bathwater quality in
terms of fecal coli and total coli. Unfortunately, this paper does not allow any scientifically
sound conclusions. The effectiveness of filters planted with A. calamus and P. australis
was compared by using findings from various years; the state of planting was not
characterized, and no unplanted control was used by way of comparison.
Rivera et al. (1995) confirmed the effect of the increased elimination of bacteria (E.
coli) in the rhizosphere by Phragmites and Typha (35 –91%) compared to unplanted
controls in microcosm investigations. However, no significant efficiency differences were
found between the two plant species (Phragmites and Typha). During the pilot-scale tests,
a lower rate of elimination was noted in the winter than in the summer, although no
significant differences were observed between the planted and unplanted variants.
In an Austrian study, the disinfection parameters were characterized in three vertically
charged constructed wetlands (Mitterer, 1995). In terms of colony-forming units, elimina-
tion was in the order of between three and four orders of magnitude. The rates of elimination
for fecal coliforms and enterococci were 3– 4 and 2 –3 orders of magnitude, respectively.
Kadlec and Knight (1996) listed the efficiency of the elimination of coliforms and
streptococci in various systems of constructed wetlands. As a rule, more than 90% of the
coliforms and more than 80% of the fecal streptococci were eliminated.
Hagendorf and Hahn (1994) studied the efficiency of a number of wetlands in
Germany. They observed the best results in systems with a mixture of sand and gravel
and vertical flow. Horizontal systems were by no means as efficient, although those with
fine to medium sandy soil afforded better germ reduction than those with pebbly soil.
However, systems with small-grained soils often resulted in hydraulic problems (clogging
leading to surface flow), which drastically reduced efficiency.
The findings of Thurston et al. (1996) regarding the comparison of a pond system with
a subsurface flow planted soil filter are very interesting, and are listed in Table 2.
As expected, the planted soil filter is more efficient at eliminating bacteria than the
Lemna pond. Surprisingly, however, the protozoa were better eliminated in the pond than
in the soil filter.
Rivera et al. (1995) also observed that the elimination of pathogens cannot be solely
explained by filtration effects. Whereas amoebae in a gravel filter were removed to a
degree of 95% (tropical climate) or 75% (subtropical climate), the efficiency of the soil
filter was only 15 – 17%.
U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117 111
Table 2
Comparison of the efficiency of a pond and a planted soil filter for wastewater disinfection (Thurston et al., 1996)
(elimination efficiency in %)
Lemna pond Planted soil filter
Giardia cysts 93 83
Cryptosporidium oocysts 91 67
Total coliforms 54 99
Fecal coliforms 59 98
Coliphages 35 94
11. Outlook
Constructed wetlands have been used to treat wastewater ever since the pioneering
work performed by Käthe Seidel in the 1960s.
Over the years, numerous examples have shown that this technology is suitable for
treating both municipal sewage and a broad range of industrial wastewater. It also gave
birth to the idea of using trees to remediate contaminated aquifers near the surface.
Whereas, originally, merely small wetlands were constructed which could only treat the
wastewater produced by a small population, in the meantime, larger treatment works have
also been built that are able to cope with a population equivalent of several thousands.
Therefore, the consequences of using this technology should be examined, such as the
technological limits of wetland size and the problems which are to be expected with large
constructed wetlands. In the past, for example, sewage farms (farmland irrigated with
wastewater) were used, which resulted in the accumulation of pollutants over a long
period. This problem can be minimized by the separate treatment of industrial and
municipal wastewater accompanied by careful monitoring. Another question that needs
to be answered is the extent to which the plant biomass in such large wetlands can
subsequently be put to viable economic use, for example, as a source of energy or as raw
112 U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117
material for the paper industry, etc. These are issues that will naturally vary from one
country to the next depending on the various socioeconomic and climatic conditions that
apply.
Given the pressing need for clean, disinfected water in the ‘Third World,’ this is an area
where the usage of constructed wetlands to solve water and wastewater problems could be
especially widespread. This is a specific field which needs further research and develop-
ment work.
In future, once constructed wetlands become better known, increasing attention is likely
to be paid to developing suitable combinations of different technologies. For example, one
energy-saving strategy would be the combination of anaerobic fermentation and post-
treatment in constructed wetlands.
Despite the experience which has been built up in years of practical application and
research, a number of fundamental aspects of exactly how constructed wetlands function
are not yet adequately understood. One reason for this is that, compared to other
technologies such as activated sludge, constructed wetlands depend on the interaction of
many more different components.
The basic aspects upon which more work is essential include:
Achieving a better understanding of the complex interactions involved will enable the
basic scientific aspects to be optimally combined with the technical possibilities available,
thus enabling wetland technologies to be used on a broader scale.
U. Stottmeister et al. / Biotechnology Advances 22 (2003) 93–117 113
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