Treatment of Contaminated Water by Constructed Wetlands: an old Technology revisited
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Lecture series Chemicals in the Environment
Treatment of Contaminated Water by
Constructed Wetlands: an old
Technology revisited
Hermann J. Heipieper
Department of Environmental Biotechnology
Helmholtz Centre for Environmental Research – UFZ
Leipzig, Germany
Leipzig, 16 October 2012
Constructed Wetlands for
Sewage and Wastewater treatment
Sewage Irrigation Fields
Were used in Europe for the treatment of domestic sewage since the
end of the 19th century and were substituted by modern waste water
treatment plants after the second world war
Renaissance of Constructed Wetlands
Renaissance of Constructed Wetlands
Parking areas along the
German Autobahnen
„Hundertwasserhaus“ in
Vienna
Comparison of constructed wetlands with other
wastewater treatment technologies
Trickling Filter Fixed Bed Reactor
Activated Sludge Systems Constructed Wetlands
Comparison of investment and running costs of constructed wetlands and 3
other wastewater treatment technologies
Costs / PE (€)
Investment
4 PE 8 PE 20 PE 50 PE
Treatment Plant Size
Trickling Filter
Fixed Bed Reactor
Activated Sludge Systems
Constructed Wetlands
Running Costs
/ PE*year (€)
4 PE 8 PE 20 PE 50 PE
Treatment Plant Size
PE = Population equivalent or unit per capita loading is the number expressing the sum of the
pollution load produced during 24 hours by industrial facilities as the pollution load in household
sewage produced by one person. One PE unit equals to 54 grams of Biochemical Oxygen Demand
(BOD) per 24 hours
What are the chemical,
physical and
biological processes
in planted systems?
Wetlands in nature and for
wastewater treatment
Sun phytovolatilization
Plants
volatilization
phytoaccumulation
☺☺
contaminants
☺☺ adsorption
☺☺ biodegradation ☺☺ filtration
Rhizosphere microorganisms
Constructed Wetlands (CWs):
Role of plants
Input of organic compounds
and O2 into the rhizosphere
uptake, assimilation aerobic zone
accumulation and metabo-
lisation of inorganic ions Micro- rhizodeposition products
organisms
Role of soil matrix SRB
Corg. + SO42- → CO2 + S2- anaerobic zone
adsorption, ion exchange
S2- + Me2+ → MeS↓
filtration, surface for biofilms
Role of microorganisms Sediment
degradation/transformation of organic and inorganic compounds
In the rhizosphere, density and diversity of microorganisms is
10 to 100-times higher than in the surrounding bulk soil
The rhizospheric effect
Aerobic conditions
S- + 2 O2 = SO42-
rhizospheric
effects
Anaerobic conditions
S- + metals (e.g. FeS)
black precipitationCWs with their macro- and micro-gradients
macro- and microgradients
Page 11Mosaic structure of aerobic and anaerobic
micro-niches in the root zones
Helophytes
Electron acceptor Electron donor
(oxygen) (rhizodeposition products,
root exudates)
The actual parameters (rH, oxygen concentration etc.) do not reflect the
actual net flow of oxygen and rhizodeposition products!
Influencing factors: Plant species
Climatic conditions (sun, temperature)
Water composition (organics, electron acceptors, pH,
buffer capacity)
Hydraulic conditions (stagnant or flowing pore water)Aerenchyma of helophytes
e.g. Juncus effusus
biologie.tu-dresden.de
With the help of these air-filled cavities in their stems, which allows
exchange of gases between the shoot and the root, helophytes are
able to provide their submerged hypoxic roots with oxygenCatabolic activity in the rhizosphere
Plants provide the rhizosphere with organic
compounds such as: alcohols, organic acids,
sugars, amino acids
Courtesy of Alvaro Gonzalias
• supply carbon source
• stimulate bacterial degradation
Transport of oxygen to the roots allows aerobic
degradation
Activities only with active photosynthesisBioremediation space
Nutrients, electron acceptors O2
Micro/macro geography
Microorganisms
Adapted from: de Lorenzo V., Curr. Opin. Biotechnol. 2008. 19:579-589Sensors for
T, rH, O2
Water jacket
T=6 °C
Magnetic
stirrer
Laboratory system for
measuring the oxygen input
91.6
Max. Oxygen Release Rate [mg h -1 plantlet -1 ]
1.4
TyphaT.
latifolia
latifolia
1.2
J.effusus
Juncus effusus
1
0.8
0.6
0.4
0.2
0
-300 -200 -100 0 100
Eh [mV]
Maximum oxygen release rates of individual experiments with several plantlets
of T. latifolia and J. effusus depending on the corresponding redox potential
(Wießner et al. 2002. Int. J. Phytorem. 4, 1-15)Oxic-anoxic interfaces and gradients
O2-gradients
Capillary fringe in soils Root zone of plants
Microbial matsWastewater Treatment in Constructed Wetlands Advantages Disadvantages • low initial costs • sensitive system • low running costs • high demand on ground • provision of habitats • seasonal fluctuations • construction with local • need of biological research materials
Challenges of Wetland Research at the UFZ
Hygienisation
(UBZ, UBT)
Generation of
climate-relevant
gases
(ANA, BOPHY, HDG, MET)
Technology
C and S sequestration development
(ANA, CATHYD) together
with UBZ
Removal of recalcitrants
of low concentration UBZ - Langenreichenbach
– pharmaceuticals …
(ANA, UBZ, ECOTOX, GWS)
Costs (Root mat filter UBZ - Langenreichenbach
technology)Investigations
in the
CW Pilot Plants
in Leuna and Bitterfeld
Zhongbing Chen, Eva Seeger, Mareike
Braeckevelt, Shubiao Wu, Oksana
Voloshchenko et al.Types of ponds/constructed wetlands
A
D
B
E
C
A: pond with floating plants
B: pond with submersed water plants
C: pond with emersed waster plants different intensities
D: CW, horizontal subsurface-flow of wastewater to air
E: CW, vertical flow and root contact!
3Constructed Wetlands for treatment
contaminated groundwater in Leuna
phytovolatilization
deep volatilization
groundwater
unsaturated
saturated zone oxic
anoxic anoxic
O2
groundwater locally
flow oxic .
mO2 ~ 10 g m-2 d-1Constructed Wetlands for treatment
contaminated groundwater in Leuna
Phragmites australis
m
5
M. Kästner, 2007
1.1 m
SEITE CITE Programmtag 2010Degradation of Benzene in the
Pilot Plant Leuna
A1
A6
A2 A3
A5
A1
A2
A4
25,000
20,000
Benzol [µg/l]
15,000
• (A1) horizontal flow planted
10,000 and unplanted soil filters
5,000 • Benzene degradation only
0
occurs in the planted soil
2.5.09 12.5.09 22.5.09 1.6.09 11.6.09 21.6.09 1.7.09 filter
Date
Outflow unplanted soil filter Outflow planted soil filter InflowMass balance
87.8%
benzene
85.7% 88.8%
9.1%
8.1%
11.2% 2.1%
3.1% 3.1%
24.2%
1.0%
43.5% 18.0% 56.5% 37.3%
MTBE
32.3%
38.5% 5.0% Plant root mat
38.5%
6.2%
worst-case scenario best-case scenario data-based scenario
Microbial degradation
}
Surface volatilization
Phytovolatilization
Accumulation, Plant uptake
Phytodegradation
Seeger et al. 2011,
EST 45, 8467-8474Mass balance
87.8%
benzene
85.7% 88.8%
9.1%
8.1%
11.2% 2.1%
3.1% 3.1%
1.0%
24.2%
43.5% 18.0% 56.5% 37.3%
MTBE
32.3%
38.5% 5.0% 38.5%
6.2%
Plant root mat
worst-case scenario best-case scenario data-based scenario
Microbial degradation
}
Surface volatilization
Phytovolatilization Seeger et al. 2011,
Accumulation, Plant uptake EST 45, 8467-8474
PhytodegradationMass balance
87.8%
benzene
85.7% 88.8%
9.1%
8.1%
11.2% 2.1%
3.1% 3.1%
1.0%
24.2%
43.5% 18.0% 56.5% 37.3%
MTBE
32.3%
38.5% 5.0% 38.5%
6.2%
Plant root mat
worst-case scenario best-case scenario data-based scenario
- very low emissions of biodegradable volatile compounds (benzene)
Microbial degradation
- high emissions of hardly degradable compounds (MTBE)
}
Surface volatilization
Phytovolatilization
Seeger et al. 2011,
Accumulation, Plant uptake EST 45, 8467-8474
PhytodegradationNitrogen transformation processes
CWs have favorable
conditions
for partial nitrification and
anammox
Canfield et al. 2010. Science. 330:192-196Investigations
in the
CW Pilot Plant
in Langereichenbach
Otoniel Carranza, Monika Möder, Jaime Nivala,
Luciana Schultze-Nobre et al.UBZ ecotechnology research facility
“Langenreichenbach”
Modified from Headley et al. (2011)
Test TEST
TES11Selected substances
Substance Use Log inlet Structures
Kow (µg L-1)
Galaxolide Personal care product 5.7 4.27 ± 4.42
Tonalide Personal care product 5.7 0.48 ± 0.47
Triclosan Personal care product 4.76 1.59 ± 0.46
Carbamazepi Pharmaceutical 2.45 11.85 ± 12.06
ne
Caffeine Pharmaceutical 0.16 31.77 ± 30.02
Ibuprofen Pharmaceutical 3.97 30.96 ± 10.13
Diclofenac Pharmaceutical 4.5 9.55 ± 3.47
Naproxen Pharmaceutical 3.18 3.17 ± 0.87
Ketoprofen Pharmaceutical 3.12 4.90 ± 1.06
Techn. Phenolic estrogenic disruptor 4.5 5.52 ± 2.47
Nonylphenol chemical
Bisphenol A Phenolic estrogenic disruptor 3.64
Test TEST
6.63 ± 7.11
chemical TES
Carranza, O. et al. 2012Sampling approach
75.0 %
50.0 %
25.0 %
12.5 %
12.5 cm
50.0 cm
Modified from Hijosa-Valsero et al. (2010)
Test TEST
TESAnalytical procedure
SPE
Multicomponent GC-MS for the quantification of pharmaceuticals
1. Filtration
Neutral
2. pH conditioning
Galaxolide
Tonalide
3. Extract sample Caffeine
through cartridge Carbamazepine
4. Drying cartridges + derivatization
Ibuprofen
5. Elution with Methanol Naproxen
Triclosan
Bisphenol A
Ketoprofen
6. Evaporation to 1 mL
Diclofenac
Techn. Nonylphenol
7. Cleanup if needed Test TEST
Carranza, O.TES
et al.132012Overall removal efficiencies
0.16
3.12
3.18
3.97
4.5
5.7
2.45
4.5
4.76
3.64
5.7
Test O.TEST
Carranza, et al. 2012
TESPOF III: Controlling Chemicals’ Fate
Investigations
in the
Planted Fixed Bed
Reactors
PFR
Zhongbing Chen, Arndt Wiessner, Uwe
Kappelmeyer, Paula Martinez, Luciana Schultze-
Nobre, Vianey Marín Cevada et al.UFZ - Phytotechnicum
Linking water and soil research:
Multiphasic process analysis in Planted Fixed-
Bed Reactors (PFR)Experimental tools: Macro-gradient free test system
for simulating root-near processes
e.g.:
- Routes and interrelations of
C, N and S transformations
- Sulphur immobilization
stability
- Toxicity effects
- Toluene degradation
(Planted Fixed Bed Reactor - PFR)
(Kappelmeyer, U. et al. 2002)Planted Fixed Bed Reactors PFR
Planted Fixed Bed Reactor with Juncus effusus
200
150
Oxygen (µg/L) / Redox Potential (mV)
100
50
0
04.07.05
05.07.05
06.07.05
07.07.05
08.07.05
09.07.05
10.07.05
11.07.05
12.07.05
-50
-100
-150
-200
Diurnal oscillation
of O2 availability -250
Redox [mV] Oxygen [µg /L]
Wießner et al. 2005. Water Res. 39:248-256.Is the catabolic activity of bacteria changing
according to oxygen and redox fluctuations?
z
z z z
z z
z
zz z
zz
Courtesy of Dr. Marcell NikolauszPFR 5 and 6 planted with Juncus effusus
PFR 6 PFR 5
Benzoate
+
TolueneToluene concetration (mg/L)
27
.A
0
5
10
15
20
25
30
35
pr
.
2.
M
ai
.
R6 In
R5 In
7.
M
ai
.
12
.M
R6 Out
R5 Out
ai
.
17
.M
ai
.
22
.M
ai
.
27
.M
Date
ai
.
1.
Ju
n.
6.
Ju
n.
11
.J
un
.
16
.J
un
.
PFR 5 and 6 planted with Juncus effususPFR 5 Deltaproteobacteria
"Bacteroidia"
"Planctomycetacia"
Illumina sequences
Holophagae 16S rRNA gene (V5-V6)
Chloroplast
Subdivision3
Chlamydiae
"Sphingobacteria"
Clostridia
Proteobacteria (70-80%)
Actinobacteria
Alphaproteobacteria
• Most abundant phylum:
Gammaproteobacteria
Betaproteobacteria
• Some diversity differences
between the two PFRs
PFR 6 Deltaproteobacteria
"Bacteroidia"
"Planctomycetacia"
Holophagae
Chloroplast
Subdivision3
Chlamydiae
"Sphingobacteria"
Clostridia
Actinobacteria
Alphaproteobacteria
Gammaproteobacteria
BetaproteobacteriaPossible Target enzymes in PFR
TOL (Xylene monooxygenase) Baldwin et al.2003
TOD (Ring hydroxylating dioxygenase) Baldwin et al., 2003
Benzyl-succinate synthase
COO-
PHE (Phenol hydroxylases)CHBaldwin
3 et al., 2003
bssA
CH
TMO (Toluene monooxygenase) Hendrickx et al., CH
2006
2
COO-
BssA (Benzyl-succinate synthase)
Howard Junca, personal
communication
Adapted from Kahng et al., 2001CATABOLIC ARRAY
Tol plasmid-like
Xylene MO
bcrA T. aromatica
Catechol-2,3-
bssA
dioxygenase G4
B. Magnetospirillum
vietnamensis sp.
TS6
Catechol-2,3-
dioxygenase
Soluble diiron MO
Vilchez-Vargas et al. 2012. Environ. Microbiol. in press.The catabolic array indicates that monooxygenation
seems to be the predominant mechanism for aerobic
toluene degradation in PFRs
Only in PFR 6, the anaerobic toluene degradation
seems to have a significant contribution
How can we confirm these results?....
...the old fashion way: IsolationAEROBIC ISOLATES
Pseudomonas sp. AET-5-01 CH3 CH2OH
Isolated from PFR5
(Baldwin BR., et al., 2003)
CH3 CH3
Ralstonia pickettii AET-6-14
Isolated from PFR6
OH
(Hendrickx B., et al., 2006a,b)
CH3 CH3
OH
Pseudomonas sp. AET-6-18 H
H
Isolated from PFR6 OH
(Hendrickx B., et al., 2006a,b)Screening with degenerate primers using cDNA
as template
Note: The cDNA was prepared using the reverse primer of each primer pair
because with hexamer primers we couldn’t detect get any PCR product
BssA (benzylsuccinate synthase) Most of the DNA bssA
-357 clones were very
-357
-134
-134
256
247
-79
247
256
mV
-79
-14
R6
-14
87
45
R5
45
87
C+
75
75
closely related to
C-
Magnetospirillum sp.
TS-6 (around 90 from
93 clones)
RT+ RT- RT+ RT-
R5 R6 C+:T aromaticaWhere do we go from here?
UBT papers:
POF III: Controlling Chemicals’ Fate (CCF)
Towards eco-compatible chemicals:
Managing ecosystem chemodynamicsLinking water and soil research:
Multiphasic process analysis in Planted Fixed-
Bed Reactors (PFR)
Temporal Fluctuations Microbial Adaptations Microbial Community & Catabolism
- Metagenomics - Lipidomics (UBT) - Metagenomics (UBT, HZI)
(UBT, HZI) - Transcript Analysis (UBT) - Protein SIP (PROTEOM)
- 2D Isotope Fractionation (ISOBIO)
- Physiology (UBT)
- Isolation (UBT)
Pollutant
e.g. toluene as a
model compound;
pharmaceuticals, rhizospheric
health care products effects
Wießner et al. 2005.
Water Res.Investigations planned with the planted fixed-bed reactors (PFR) within Workpackage 2 of CCF (POF3) Degradation of a pollutants such as toluene or e.g. ibuprofen or paracetamol in at least two PFRs. Elucidation of the microbiota responsible for the degradation under aerobic and anaerobic conditions. Analysis of two-dimensional isotope fractionation in order to elucidate the catabolic pathways. Detection of the catabolic genes responsible for the degradation. This will be done by metagenomic and clone library techniques as well as by applying a catabolic gene array developed by the HZI. The expression of these catabolic genes will be systematically analyzed under seasonal (winter-spring-summer-autumn) as well as diurnal (day- night) cycles. Addition of the compounds in form of the 13C-labelled stable isotopes. Monitoring of the development of microbial and catabolic diversity by protein-based stable isotope probing (protein SIP).
Potential pharmaceuticals to
be investigated
Paracetamol
IbuprofenMore technical activities
Future Research Questions for CWs
Effect of filter material additives gravel, + activated carbon, + ferric iron
Performance efficiency of CW types gravel filter system vs. plant mat
Integ. Contaminant massbalance microbial degradation, plant uptake, emission
Technical approach for promoting removal efficiency
Filter material
additives: Plant mat without gravel Mass balance incl. emissionConstructed wetland efficiencies - Benzene
Comparison of wetland type: planted gravel filter vs. plant mat
planted filter bed
plant mat
100
80 Plant mat without gravel
residual load [%-inflow]
60
benzene
40
20
0
1.6.07 1.10.07 1.2.08 1.6.08 1.10.08 1.2.09 1.6.09
Seeger et al. 2011, Environ. Poll. 159, 3769-3776Nitrogen transformation processes
N2
NH4+
Tools: [NH2OH]
Isotope fractionation,
S0
FISH, qPCR
NO
N 2O
NO NO2-
NO2- NH4+
NO3-
CWs have favorable conditions H 2S
for partial nitrification and anammox CorgRemoval mechanisms of pathogenic germs in CWs
Hygienisation
Protozoa
Bacteriophages
Bdellevibrio
Plants
lawrence-throughthevisor.blogspot.com
Main future applications of CWs will be the treatment of
domestic sewage in countries such as China, India,
Vietnam, etc.
EU project: Water4cropsAcknowledgements
Peter Kuschk
Arndt Wiessner
Uwe Kappelmeyer
Jochen A. Müller
Matthias Kästner
Paula Martínez Lavanchy
Zhongbing Chen
Vianey Marín Cevada
Luciana Schultze Nobre
Mareike Braeckevelt
Eva M. Seeger
Alexander Al-Dahoodi
Helga Fazekas
Otoniel Carranza
Jaime Nivala
ANA/UBT and many more colleagues!
Fabio Kraft
Anja Taubert
Ilja Iwlew
Claudia Pietsch
Kerstin Puschendorf
Ines Mäusezahl
Kerstin Ethner
Ivonne Nijenhuis ISOBIO
Jana Seifert PROTEOM
Monika Möder ANA
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