Environ Sci Pollut Res
DOI 10.1007/s11356-014-3247-3
WETLAND SYSTEMS: ECOLOGY, FUNCTIONS AND MANAGEMENT
Screening of 18 species for digestate phytodepuration
Francesca Pavan & Simone Breschigliaro &
Maurizio Borin
Received: 2 February 2014 / Accepted: 23 June 2014
# Springer-Verlag Berlin Heidelberg 2014
Abstract This experiment assesses the aptitude of 18 species
in treating the digestate liquid fraction (DLF) in a floating
wetland treatment system. The pilot system was created in NE
Italy in 2010 and consists of a surface-flow system with 180
floating elements (Tech-IA®) vegetated with ten halophytes
and eight other wetland species. The species were transplanted
in July 2011 in basins filled with different proportions of DLF/
water (DLF/w); periodic increasing of the DLF/w ratio was
imposed after transplanting, reaching the worst conditions for
plants in summer 2012 (highest EC value 7.3 mS cm/L and
NH4-N content 225 mg/L). It emerged that only Cynodon
dactylon, Typha latifolia, Elytrigia atherica, Halimione
portulacoides, Salicornia fruticosa, Artemisia caerulescens,
Spartina maritima and Puccinellia palustris were able to
survive under the system conditions. Halophytes showed
higher dry matter production than other plants. The best root
development (up to 40-cm depth) was recorded for
Calamagrostis epigejos, Phragmites australis, T. latifolia
and Juncus maritimus. The highest nitrogen (10–15 g/m2)
and phosphorus (1–4 g/m2) uptakes were obtained with
P. palustris, Iris pseudacorus and Aster tripolium. In conclu-
sion, two halophytes, P. palustris and E. atherica, present the
highest potential to be used to treat DLF in floating wetlands.
Keywords Anaerobic digestion wastewater . Floating
wetlands . ECw . Salt tolerance halophytes . Nutrient removal
Responsible editor: Thomas Braunbeck
F. Pavan (*) : S. Breschigliaro : M. Borin
Department of Agronomy, Food, Natural Resources, Animals and
Environment (DAFNAE), University of Padua, Agripolis Campus,
Viale dell’Università 16, 35020 Legnaro, PD, Italy
e-mail: francesca.pavan.6@studenti.unipd.it
M. Borin
e-mail: maurizio.borin@unipd.it
Introduction
In recent years, as a consequence of high renewable energy
subsidies, anaerobic digestion (AD) of biomasses and animal
manures has spread throughout Europe, and many installa-
tions have recently been constructed (EurObserv’ER 2010). In
Italy, the number of anaerobic digesters has risen from 587 in
2011 to 994 in 2012 with a power increase of 116 % (CRPA
2012). In 2012, the electrical power installed was 765 MW.
The average installed power of these plants is 0.5 MWel with
an average daily digestate production of 100 m3 per installed
MWel (Balsari et al. 2013), corresponding to an estimated
35,000–40,000 m3/day of digestate.
Anaerobic digestion is an environmentally sustainable way
to manage organic waste (Appels et al. 2008; Tambone et al.
2010). No less important are the effects of AD on the envi-
ronment, which contribute to reducing global warming, not
only by replacing fossil fuel with biogas but also by carbon
storage in the soil and inorganic fertilizer substitution (Møller
et al. 2009). In fact, with AD, mineral fractions of N and P are
enriched during digestion (Massé and Droste 2000), resulting
in a higher concentration of plant-available nutrients com-
pared to undigested organic material, promoting plant growth
in a similar way to the use of mineral fertilizers (Appels et al.
2008; Gutser et al. 2005; Liedl et al. 2006).
The application of organic effluents to agricultural soils is
regulated by the Council Directive 91/676/EEC (1991), which
limits the amount of nitrogen that can be spread on farmland.
These limits pose particular challenges that cannot be solved
just by the regulation of land spreading. Even if biogas pro-
duction systems allow the exploitation of livestock effluent as
a source of income, they do not diminish the nitrogen load. In
fact, AD mineralizes a part of the slurry organic nitrogen in
ammonia nitrogen and favours more bio-available nitrogen
forms, such as ammonium (Franchino et al. 2013). In addition,
salt solubilization and mineralization of organic matter during

AD processes increase soluble salt concentration in digestate
(Massaccesi et al. 2013). To facilitate the management and the
agricultural use of this by-product, the digestate is separated
into liquid (DLF) and solid fractions, with different organic
matter and ammonia nitrogen compositions.
In areas where the nitrogen load coming from digestate
exceeds the amount that can be spread on fields, adequate
treatments have to be adopted. Constructed wetlands (CWs),
with advanced system layout, could offer an efficient and
environmentally valuable approach to the treatment of
digestate liquid fraction (DLF) (Comino et al. 2013), because
they are effective in removing biodegradable organic matter,
as well as residual suspended solids, and can also remove
nutrients (nitrogen and phosphorus) (García 2004; Kadlec
et al. 2000; Soto et al. 1999).
Constructed wetlands with free water surface (FWS CWs)
have been used for many purposes worldwide. It has been
reported that treatment performance of planted FWS CWs is
superior to unvegetated basins (Vymazal 2013). However,
treatment performance of FWS CWs could be affected by
the plant species used (Vymazal 2013).
FWS CWs can be improved by installing floating elements
that allow the use of a greater number of species. Floating
treatment wetlands are artificially created systems using var-
ious ways to trap macrophytes in self-buoyant mats (De
Stefani et al. 2011). The plant roots hanging beneath the
floating mat provide an extensive surface area for attached
biofilm growth and entrapment of fine suspended particulates.
Because the plants are not rooted in soils in the base of the
wetland, they are forced to acquire their nutrition directly from
the water column, which may enhance nutrient and element
uptake rates into the biomass (Tanner and Headley 2011).
Floating systems developed around the world have performed
well in pollution control (removing suspended solids (SS),
nitrogen (N), phosphorus (P), biochemical oxygen demand
(BOD), chemical oxygen demand (COD) and metals in par-
ticular) and in the creation of new water environments (De
Stefani et al. 2011). Floating treatment wetlands have poten-
tiality but there is a need for further pilot studies and field-
scale testing and evaluation on a range of water treatment roles
(Tanner and Headley 2011) and plant species that can be used.
A total of 35 species were recorded in constructed wetlands
in Europe (Vymazal 2013), but only some of them have been
studied in floating systems, and no information is available on
DLF treatment. Furthermore, the digestate electrical
conductivity reduces the range of species that can be used
for this purpose.
As Webb et al. (2013) underline, considerable research has
been carried out on systems planted with glycophyte plant
species for remediation of freshwater effluents, which is not
directly transferable to saline systems. There is growing inter-
est in the potential of constructed wetlands planted with fac-
ultative or obligate halophytes for the remediation of saline
Environ Sci Pollut Res
effluent and marine intensive land-based aquaculture waste-
waters (Brown et al. 1999; Calheiros et al. 2012; Lin et al.
2002, 2003, 2005; Lymbery et al. 2006, 2013; Sousa et al.
2011; Webb et al. 2012) or, as in this case, to treat the by-
product of AD that presents comparable conductivity to
seawater.
Previous research on CWs planted with emergent macro-
phytes (e.g. Phragmites spp., Typha spp.) for the treatment of
saline/brackish wastewater is limited (Klomjek and
Nitisoravut 2005; Lin et al. 2002, 2003, 2005; Lymbery
et al. 2006), and there are even fewer studies on the use of
halophyte planted CWs for saline wastewater remediation
(Brown et al. 1999; Brown and Glenn 1999; Calheiros et al.
2012; Sousa et al. 2011). It is therefore worth increasing
knowledge on the use of halophyte species in constructed
wetland systems.
The aim of this study is the screening of 18 species,
including ten halophytes, planted in floating elements in a
surface-flow wetland system treating the digestate liquid frac-
tion. Their survival potential in DLF, their adaptability to
floating systems and their aptitude to depuration have been
evaluated.
Material and methods
A pilot FCW system was constructed in 2010 in north-eastern
Italy to treat the nitrogen load of one anaerobic digester’s
effluents. It consists of three separate square pools with a
water depth of about 0.6 m and surface areas that differ
slightly: 27 m2 in pool 1 (P1), 30 m2 in pool 2 (P2) and
33 m2 in pool 3 (P3). The pools were waterproofed by sheets
of polyolefin. Each pool surface was completely covered with
Tech-IA® floating elements: 54 in P1, 60 in P2 and 66 in P3.
The self-floating elements are produced in ethylene vinyl
acetate (EVA), rectangular (50×90 cm), with eight apertures
each of which has grids to sustain plants, but for this study, we
used only four. Six holes in the frame can be used to easily
connect the single elements to each other and to the pool sides.
Considering the surface area of one Tech-IA® and the sur-
rounding space, it can be approximated that two Tech-IA®
occupy a surface area of 1 m2.
In this survey, we compared the behaviour of 18 species.
Ten of them were collected from the Venice lagoon, not far
from the experimental site, and are halophytes; therefore, they
are expected to be able to adapt to conductivity of digestate
and waterlogging (Artemisia caerulescens L., Aster tripolium
L., Halimione portulacoides (L.) Aellen, Inula crithmoides L.,
Juncus maritimus Lam., Limonium narbonense Mill.,
Puccinellia palustris (Seen.) Hayek, Sarcocornia fruticosa
(L.) A. J. Scott, Spartina maritima (Curtis) Fernald,
Elytrigia atherica (Link) Kerguélen). A further eight species
were chosen, some of which are generally used in constructed
Environ Sci Pollut Res

wetlands (Cynodon dactylon (L.) Pers., Alnus glutinosa L., Table 1 Species identifying code and number of species, plants and
Tech-IA floating elements in each pool
Arundo donax L., Iris pseudacorus L., Phragmites australis
(Cav.) Trin. ex Steud., Typha latifolia L., Calamagrostis ID Scientific name Number of plants
epigejos (L.) Roth and Cladium mariscus (L.) Pohl).
Transplanting was done in summer 2011 with random layout, P1 P2 P3 Total
using four plants of the same species per Tech-IA®, so about AD Arundo donax 12 12 12 36
eight plants per square metre. A variable number of Tech-IA AG Alnus glutinosa 12 12 12 36
per species were installed in the three pools, and this led to the CD Cynodon dactylon 12 12 12 36
number of plants reported in Table 1 and to a different number CE Calamagrostis epigejos 12 12 12 36
of plants in each pool but maintaining the same planting CM Cladium mariscus 12 12 12 36
density (Table 4). IP Iris pseudacorus 12 12 12 36
After transplanting, there was a short adaptation period of PA Phragmites australis 12 12 12 36
about a week in which plants lived only in water; the DLF/ TL Typha latifolia 12 12 12 36
water (DLF/w) ratio in the pools was then increased (Table 2), AC Artemisia caerulescens 12 12 12 36
reaching the highest concentration in summer 2012. AT Aster tripolium 12 16 16 44
From June to November 2011 and from March to October EA Elytrigia atherica 12 12 16 40
2012 temperature, redox potential (ORP), electrical conduc- HP Halimione portulacoides 12 16 20 48
tivity (ECw), oxygen demand (OD) and pH in the pools were IC Inula crithmoides 12 16 20 48
determined weekly with a digital electrochemical measuring JM Juncus maritimus 12 16 20 48
system (Hach Lange HQ 40d). In the same periods, water LN Limonium narbonense 12 16 16 44
samples were collected every 2 weeks from four corners of PP Puccinellia palustris 12 16 20 48
each pool in falcon tubes and analysed with spectrophotomet- SF Sarcocornia fruticosa 12 12 12 36
ric analysis (DR 2800 Hach Lange) to measure the concen- SM Spartina maritima 12 12 16 40
tration of total nitrogen (TN), ammonia nitrogen (NH4-N), Total plant number 216 240 264 720
nitrate nitrogen (NO3-N), total phosphorus (TP), orthophos- Total Tech-IA number 54 60 66 180
phate (PO4-P), chemical oxygen demand (COD) and turbidity
(NTU).
The phenology and behaviour of each plant were Results and discussion
monitored and recorded every week to determine mor-
tality and senescence. To assess the survival rate, the Wastewater quality
number of living plants in November 2011, in March
and in October 2012 was compared with the number in Wastewater characteristics (Table 3) allowed to describe the
the previous period. conditions in which plant roots lived during the study period.
In November 2011 and October 2012, the plant above mat The analysis showed statistically significant differences
biomass was harvested, and dry matter production of each among pools only for ammonia nitrogen and total N.
floating element was measured. Above mat dry matter of each Ammonia nitrogen concentrations were higher in P1 (median
species harvested from all pools was mixed, and a sample was value 110 mg/L) than in P2 (median value 72.4 mg/L) and P3
collected to determine total Kjeldahl nitrogen (TKN) and (median value 64.1 mg/L); total nitrogen in P1 (median value
phosphorus (TP) content, using the FAO method (FAO 134 mg/L) differed from P3 (median value 93.8 mg/L). In
2011). At the end of the growing season 2012, root measure-
ments were taken; for each plant, root depths and four root
system diameters were measured: near the crown, at mid- Table 2 Date and
amount (m3) of DLF in- DLF input in pools (m3)
depth, at 3/4 and at the end of root system. put in pools
The data collected by monitoring the effluent for each of Date P1 P2 P3
the three pools were compared using the Kruskal-Wallis test to July 12, 2011 1 1 1
identify statistically significant differences. This test is an August 23, 2011 1 1 1
alternative to the F test analysis of variance, which is used September 13, 2011 1 0,3 0
for studies of simple classification where various groups are March 22, 2012 0 0,7 1
compared using the median values. In all cases, the differences May 23, 2012 0 0 1
between the medians were considered statistically significant June 08, 2012 1 1 1
at p value of 0.05 or less. The statistical analysis of the data June 26, 2012 1 1 1
was performed using the R software, operating in Windows m3 tot 5 5 6
environment.
Table 3 Monthly median, first and third quartile values of wastewater parameters measured in the pools in 2011 and 2012

NO3-N NH4-N* TN** COD TP Eh

mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mV mV mV

Month/year Quart 1 Median Quart 3 Quart 1 Median Quart 3 Quart 1 Median Quart 3 Quart 1 Median Quart 3 Quart 1 Median Quart 3 Quart 1 Median Quart 3
June 2011 0.5 0.6 0.8 0.1 0.1 0.1 10.4 13.4 28.4 110.0 116.0 133.0
July 2011 2.2 3.5 5.1 12.8 34.1 45.0 30.7 55.9 78.5 467.8 692.0 1064.5 7.6 9.3 10.0 −169.0 −57.8 −28.0
August 2011 3.9 5.4 7.1 31.2 43.6 54.9 51.7 71.4 92.3 578.0 887.0 1025.0 7.6 10.6 13.4 −165.3 −99.9 −25.0
September 2011 10.0 12.3 14.0 76.2 91.1 100.2 150.0 156.0 192.0 1857.0 2085.0 2234.0 22.6 23.6 24.1 −181.7 −140.1 −118.5
October 2011 12.5 13.9 16.4 79.4 85.5 100.0 141.3 152.5 174.5 1740.3 2066.5 2234.5 22.3 25.4 25.9 32.0 94.5 153.5
November 2011 10.0 10.9 13.4 73.4 102.0 117.0 115.0 152.0 179.0 1454.0 1594.0 1923.5 20.8 21.5 24.3 177.0 177.0 177.4
March 2012 5.9 8.8 10.4 57.0 85.1 105.2 20.2 71.3 109.0 797.0 1163.0 2173.0 11.1 11.7 12.5
April 2012 12.2 13.1 14.4 84.0 99.5 138.5 112.8 131.0 181.5 1820.5 1962.5 2466.0 23.7 24.6 25.4 −264.2 −218.0 −142.6
May 2012 10.4 10.9 11.1 65.6 71.0 75.6 114.8 117.5 121.0 1710.8 1784.0 1861.8 17.0 19.5 21.5 −200.2 −158.2 −133.1
June 2012 17.0 17.7 18.4 156.5 162.0 169.0 215.0 223.0 227.0 2595.5 2695.0 2812.5 26.9 27.5 28.2 −272.1 −210.6 −173.6
July 2012 13.2 17.9 21.6 165.0 193.0 205.0 219.0 236.0 262.0 3437.0 3505.0 4154.0 13.2 17.3 27.3 −273.6 −266.8 −259.8
August 2012 20.4 23.0 24.8 134.5 157.0 159.5 155.5 177.0 178.0 2743.0 2916.0 2983.5 28.0 28.7 30.0 −291.7 −280.3 −255.7
September 2012 10.1 11.3 11.6 53.6 71.5 76.8 76.5 80.1 101.1 1271.5 1385.0 1449.8 16.0 17.1 19.0 −339.9 −266.9 −188.7
October 2012 10.5 12.6 13.2 63.2 85.8 100.4 80.9 105.0 126.0 1415.5 1495.0 1560.0 21.1 22.3 22.8 −418.3 −362.1 −334.7
All values 6.1 11.1 13.8 44.6 75.6 112.5 71.4 116.0 165.0 936.0 1574.0 2247.0 11.6 20.0 24.1 −237.5 −158.9 −41.7

Each value is calculated on six data, 12 for redox potential except for the last line. Asterisks indicate significant difference at p≤0.05 using the Kruskal-Wallis non-parametric test
*Significant differences at 0.005 among pools (P1a , P2b , P3b ); **significant differences at 0.005 among pools (P1a , P2ab , P3b )
Environ Sci Pollut Res
Environ Sci Pollut Res
Table 4 Plant density in each
pool at the beginning and end of Plant density (plants/m2)
each growing season and dry
above mat biomass (g/m2) in each Pool Month
pool combining all species
July 2011 Nov 2011
P1 8.00 7.26
P2 8.00 7.73
P3 8.00 7.15
particular, from July to September 2011, the values of ammo-
nia N increased, reaching concentrations lower than 100 mg/L
in P2 and P3, but exceeding 100 mg/L in P1 (Fig. 1). This
difference can be explained with the higher amount of DLF
applied in P1 during the period. In 2012, concentrations of
ammonia nitrogen were always higher in P1 and reached a
maximum of 184 mg/L in April and 225 mg/L in July. Total
nitrogen followed the ammonia nitrogen trend.
The differences among pools can be related to the interac-
tion between DLF load and vegetation, considering the overall
effect of the various species. The higher load in P1 during
2011 might have caused some damage to the plants, that
resulting in a lower capacity to overcome the winter and at
the start of the 2012 growing season, P1 had almost 40 % less
plants than P3 and 24 % less than P2 (Table 4). During the
growing season 2012, mortality of plants occurred in all the
pools, but the ranking in density (P3>P2>P1) remained. The
different plant density might have induced different N abate-
ment in the pools due to both plant uptake and microbial
activity in the root biofilm.
Total phosphorus concentrations oscillated between 6.12
and 31.2 mg/L, reaching highest values in August 2012.
ECw (Fig. 2) followed an increasing trend in the first months
of 2011, reaching a maximum of 4.38 mS/cm in P1 (October 4,
2011). At the beginning of 2012, ECw in P1 was around 4 mS/
cm, while it was half that in the other pools. From March to
May 2012, values oscillated between 3 and 5 mS/cm; at the
beginning of June until the end of August, the conductivity
increased rapidly reaching 7.3 mS/cm in P3 on July 9, 2012.
The increase may be due to the progressive addition of DLF;
the trend also corresponded with a period of increasing temper-
ature (around 25 °C) and very low rainfall (less than 20 mm/
month) that could have stimulated evapotranspiration.
Another important parameter that could be unfavourable
for plants was the availability of oxygen for roots. The anal-
ysis of redox potential (ORP) indicated that the values in 2011
varied between 50 and −300 mV; values around −50 mV
indicated anoxia and were favourable to denitrification. In
2012, anaerobiosis conditions prevailed in the pools; the
values were always under −50 mV; high negative values as
in this case could contribute to denitrification, polyphosphate
degradation and production of sulphide, acids and methane
(Masotti and Verlicchi 2009).
Above mat dry biomass g/m2/year
Month
Mar 2012 Oct 2012 Nov 2011 Oct 2012
2.22 1.59 177 149
2.93 2.33 197 192
3.64 2.73 193 208
These values were due to the large amount of undecom-
posed organic matter supplied with DLF input in pools and
expressed by the COD values. Indeed COD values rose after
DLF input, reaching maximum values in September 2011
(2,306 mg/L) and July 2012 (5,081 mg/L).
Plant survival
From transplanting to November 2011, almost all species
showed a survival rate of 70–100 % (Fig. 3); the lower values
were recorded for A. glutinosa, C. epigejos, L. narbonense,
S. maritima and (only in P 1) T. latifolia. The DLF/w ratio
was not yet too critical; indeed there were only three applica-
tions of DLF before November. The generally good survival
might be due also to healthy storage organs of plants (coming
from a nursery) that supplied nutrients and limited the interac-
tions with the DLF-w mixture. Although the amount of DLF/w
was higher in P1, the three pools presented similar survival
capacity (Table 4).
From November 2011 to March 2012, the species showed
different ability to survive winter conditions. Almost all the
plants of A. donax, A. glutinosa and A. tripolium died; con-
versely, S. maritima had survival of 100 %, followed by
P. palustris (93 %), T. latifolia (78 %), A. caerulescens
(78 %) and P. australis (72 %) (Fig. 3, Table 5). The mortality
has to be related to the effects of cold during winter; in fact,
even though the species used were indigenous and adapted to
the climate, being placed above the floating elements without
any medium, their perennial organs had no protection against
the cold. Some species as A. caerulescens, J. maritimus and
P. australis had higher mortality in P1 probably due to higher
ammonia concentration in the autumn that could have weak-
ened the storage organs (Li et al. 2013).
The increasing values of pollutant concentration caused
mortality during the vegetative season 2012. In particular,
the higher values of ammonia in P1 could be the reason for
the higher mortality than that observed in the other two pools.
There are a lot of examples of phytotoxic effects of ammonia
concentrations of about 200 mg/L in the literature (de
Casabianca-Chassany et al. 1992; Reddy et al. 1983; Tanner
1994). This could explain the mortality of some species in
July 2012, but not the mortality of some helophytes.
T. latifolia performed better than the indications in Surrency
 Environ Sci Pollut Res

Fig. 1 Ammonia concentrations 250

in the three pools during the
monitoring period. Vertical bars 225
indicate the dates of DLF 200
distribution according to Table 2
175

Ammonia-N (mg/L)
150

125

100

75

50

25

2012-02-11
2011-06-01
2011-06-16
2011-07-01
2011-07-16
2011-07-31
2011-08-15
2011-08-30
2011-09-14
2011-09-29
2011-10-14
2011-10-29
2011-11-13
2011-11-28
2011-12-13
2011-12-28
2012-01-12
2012-01-27

2012-02-26
2012-03-12
2012-03-27
2012-04-11
2012-04-26
2012-05-11
2012-05-26
2012-06-10
2012-06-25
2012-07-10
2012-07-25
2012-08-09
2012-08-24
2012-09-08
2012-09-23
2012-10-08
2012-10-23
P1 P2 P3

(1993), who noted that it was stressed when the external
ammonia concentration averaged from 160 to 170 mg/L.
Similarly, P. australis remains unaffected at concentrations
of ammonia up to 160 mg/L and higher values (Surrency
1993).
Humenik et al. (1999) found that the tolerance to ammonia
of Juncus effusus L. and Schoenoplectus tabernaemontani
was up to 350 mg/L; even if J. maritimus is a different species,
it could explain the survival capacity in our system.
The addition of DLF induced a progressive increase in
ECw, which could be another reason for plant mortality. In
soil of the salt marshes of the northern Venice lagoon where
the halophyte species live, EC of topsoil is in a range between
6.58 and 9.80 mS/cm (Lang et al. 2010). Accordingly, only
more tolerant species should survive the conditions of our
wastewater. The results obtained from the observations
(Table 5) confirmed this hypothesis. Most halophytes sur-
vived even at the most critical moment in July, and some,
such as L. narbonense, J. maritimus and H. portulacoides,
continued their lifecycle and began flowering. Most of the
other species died, except C. dactylon, P. australis and
T. latifolia. Some of the species used in the experiment, after
a period of suffering, restarted their vegetative activity in
September 2012: some plants of C. dactylon, P. australis

Fig. 2 Conductivity values in the 8000

three pools during the monitoring
period. Vertical bars indicate the 7000
dates of DLF distribution
according to Table 2 6000
EC (µS/cm)

5000

4000

3000

2000

1000

0
11/02/12
01/06/11
16/06/11
01/07/11
16/07/11
31/07/11
15/08/11
30/08/11
14/09/11
29/09/11
14/10/11
29/10/11
13/11/11
28/11/11
13/12/11
28/12/11
12/01/12
27/01/12

26/02/12
12/03/12
27/03/12
11/04/12
26/04/12
11/05/12
26/05/12
10/06/12
25/06/12
10/07/12
25/07/12
09/08/12
24/08/12
08/09/12
23/09/12
08/10/12
23/10/12

P1 P2 P3
Environ Sci Pollut Res

Fig. 3 Survival rate of the Pool 1

species in each pool; the three 100
columns refer to three periods 90
80

Survival rate (%)

70
60
50
40
30
20
10
0
AD AG CE CM CD IP PA TL AC AT EA HP IC JM LN PP SF SM
Species

Pool 2
100
90
80
Survival rate (%)

70
60
50
40
30
20
10
0
AD AG CE CM CD IP PA TL AC AT EA HP IC JM LN PP SF SM
Species

Pool 3
100
90
80
Survival rate (%)

70
60
50
40
30
20
10
0
AD AG CE CM CD IP PA TL AC AT EA HP IC JM LN PP SF SM
Species
Jul 11 - Nov 11 Nov 11 - Mar 12 Mar - Oct 12

and T. latifolia and among halophytes A. caerulescens,
E. atherica, S. maritima and some plants of P. palustris.
Another important factor that could have affected the spe-
cies behaviour is the oxygen lack in the root zone. Anoxic
conditions of the wastewater were more endurable for macro-
phytes and for waterlogging-tolerant species, such as
S. maritima a plant indicator of anoxic conditions. Instead,
S. fruticosa and L. narbonense are indicators of oxic condi-
tions (Lang et al. 2010), and this could explain the high
mortality of these two species in our experiment.
J. maritimus, P. palustris and P. australis are characteristic
of histic soils, and this affirms their adaptability to anoxic and
eutrophic water (Lang et al. 2010).
Biomass production
The dry biomass production in 2011 (Fig. 4) was below 400 g/
m2 for all species except T. latifolia, H. portulacoides,
P. palustris and S. fruticosa. In general, biomass production in
2011 was conditioned by the nursery effect, while production in
2012 (Fig. 4) underlined more differences among the species. In
2012, the species that produced higher dry biomass were
 Environ Sci Pollut Res

Table 5 Survival rate of the species and median population values of dates of death and senescence calculated in the three pools combined

Scientific name Survival rate (%) Senescence (S) or death (D)

date median values of the
Jul 2011–Nov 2011 Nov 2011–Mar 2012 Mar–Oct 2012 % tot period Jul 2011–Oct 2012 population

Arundo donax 100 3 0 0 D: 22/06

Alnus glutinosa 67 4 0 0 D: 22/06
Calamagrostis epigejos 78 21 0 0 D: 02/07
Cladium mariscus 100 25 67 17 D: 24/07
Cynodon dactylon 89 19 83 14 S: 31/07
Iris pseudacourus 100 31 0 0 D:16/07
Phragmites australis 100 72 62 44 D:09/07
Typha latifolia 78 71 100 56 S:31/07
Artemisia caerulescens 100 78 79 61 S: 21/08
Aster tripolium 100 7 0 0 D: 09/08
Elytrigia atherica 100 80 100 80 –
Halimione portulacoides 100 50 96 48 –
Inula crithmoides 100 31 40 13 D:21/08
Juncus maritimus 100 19 78 15 –
Limonium narbonense 91 18 71 11 –
Puccinellia palustris 100 92 100 92 S:22/06
Sarcocornia fruticosa 100 17 100 17 D:10/05
Spartina maritima 50 100 70 35 S:09/07
Average rate 92 41 58 28

A. tripolium, which doubled its 2011 productivity, P. palustris that in our system, P. palustris produced 691.9 g/m2, a little
and E. atherica, all belonging to the halophyte group. A com- more than that in a study by Scarton (2006) of a plot called La
parison between the biomass production of some halophytes in Grisa in Venice lagoon, who reported that the vegetation in the
this study and the biomass production in Venice lagoon shows plot was dense and luxuriant. S. maritima had comparable

Fig. 4 Dry above mat biomass 1200

production in 2011–2012 (g/m2);
the last ten species are halophytes
1000
Above mat dry weight (g/m2)

800

600

400

200

0
CD
CE
CM

JM

CD
CE

JM
AD
AG

IP
PA
TL
AC
AT
EA
HP
IC

LN
PP
SF
SM

AD
AG

CM
IP
PA
TL
AC
AT
EA
HP
IC

LN
PP
SF
SM

Nov 2011 Species ID Oct 2012

Environ Sci Pollut Res

Table 6 N and P concentration (% on dry weight) of the different species Among the non-halophyte group, it is interesting to high-
in the 2 years in the three pools combined
light the productivity increase of I. pseudacorus from 2011 to
Species TKN (%) TP (%) 2012, confirming its suitability to be grown in floating wet-
lands (Mietto et al. 2013). Similarly, C. dactylon was more
2011 2012 2011 2012 productive in the second year, while both T. latifolia and
A. caerulescens 3.46 2.12 0.81 0.11 P. australis gave lower biomass. Their productivity was quite
A. donax 3.22 – 0.80 – low if compared to values reported in the literature, which
A. glutinosa 1.41 – 0.06 – range between 490–5,602 g/m2 for T. latifolia and 942–
A. tripolium 3.91 1.71 0.52 0.49 10,800 g/m2 for P. australis (Kadlec and Wallace 2009).
C. dactylon 4.04 3.66 0.22 0.36 Both species are able to give high production at ammonia
C. epigejos 3.75 2.14 0.20 0.27 concentration of 200 mg/L (Borin and Salvato 2012) and are
C. mariscus 2.08 2.40 0.30 0.46 suitable for floating systems (De Stefani et al. 2011; Mietto
E. atherica 3.29 2.13 0.94 0.16 et al. 2013). The low productivity can be explained as a
H. portulacoides 4.13 3.09 0.40 0.20 depressive effect of ECw since the plants used in the experi-
I. crithmoides 3.29 2.83 0.59 0.20 ment came from freshwater channels.
I. pseudacorus 2.41 2.77 0.32 0.33
J. maritimus 2.29 2.22 0.34 0.14
L. narbonense 4.08 2.49 0.24 0.20 Nutrient concentrations and uptake
P. australis 2.69 2.53 0.51 0.23
P. palustris 3.55 2.16 1.00 0.30 The concentration of TKN and TP in dry biomass was higher
S. fruticosa 3.16 – 1.19 – in 2011 than in 2012 (Table 6) except for C. mariscus,
S. maritima 2.34 2.33 1.06 0.22 I. pseudacorus and T. latifolia. This could be explained by
T. latifolia 1.88 2.04 0.08 0.25 the favourable conditions in which plants lived in 2011 and
Average values 3.05 2.44 0.53 0.26 may be an indication of the concentration that every species
might reach in better conditions. In 2011, C. dactylon,
L. narbonense and H. portulacoides dry biomass had a con-
centration of TKN higher than 4 %. The concentration was
results in 2012, but H. portulacoides, J. maritimus, lower in 2012, but C. dactylon and H. portulacoides reached
L. narbonense and S. fruticosa produced a lower quantity of the greatest concentrations among all species (higher than
biomass than in Venice lagoon. 3 %). Although these results, their uptake aptitude was

Fig. 5 Above mat biomass 30

nitrogen uptake values of each
species (g/m2) in each year; the
last ten species are halophytes
25
Nitrogen uptake (g/m2)

20

15

10

0
CD
CE
CM

CD
CE
CM

PA
AD
AG

PA
TL

HP

JM
IP

AC
AT
EA

IC

LN
PP
SF
SM

AD
AG

IP

TL
AC
AT

HP

JM
EA

IC

LN
PP
SF
SM

Nov 2011 Species ID Oct 2012

 Environ Sci Pollut Res

Fig. 6 Above mat biomass 7

phosphorus uptake values of each
species (g/m2) in each year; the
last ten species are halophytes 6

Phosphorus uptake (g/m2)

4

0
CE
CD

CM

AD

CD
CE
CM
AD
AG

IP
PA
TL
AC
AT
EA
HP
IC
JM
LN
PP
SF
SM

AG

IP
PA
TL
AC
AT
EA
HP

JM
IC

LN
PP
SF
SM
Nov 2011 Species ID Oct 2012

influenced by their ability to produce biomass in our system. 2012 despite the system conditions, instead H. portulacoides
For example, the uptake aptitude of C. dactylon increased in biomass production was noticeably affected in 2012.

Width
20 cm
Depht

Fig. 7 Root development of the studied species (cm)

Environ Sci Pollut Res
The highest nitrogen uptake values were between 10 and
15 g/m2 (Fig. 5), reached by A. tripolium, H. portulacoides,
P. palustris and S. fruticosa in 2011 and by P. palustris,
I. pseudacorus, A. tripolium and E. atherica in 2012. Very
few examples of uptake of these species are found in the
scientific literature; both T. latifolia and P. australis showed
lower uptake compared to the values reported by Kadlec and
Wallace (2009) and Borin and Salvato (2012).
Phosphorus concentrations were higher in P. palustris,
S. fruticosa and S. maritima dry biomass. Kadlec and
Wallace (2009) reported some above biomass P concentration
examples for T. latifolia, the range of values was from 0.24 to
0.42 % and is comparable with the result in our system in
2012. P. australis also gave comparable results in 2012, but
higher in 2011. In our system, considering also the biomass
production (g/m2) of the species, the highest phosphorus
values were given by S. fruticosa, P. palustris and
E. atherica in 2011 and by A. tripolium and P. palustris in
2012 (Fig. 6).
Root system
In floating mat systems, roots are directly in contact
with the water column and play an important role in
water treatment. Root biomass and pollutant accumula-
tion in roots are thus critical parameters to investigate if
the aim is to assess plant suitability to treat digestate
using floating mats. Nevertheless, the necessity to con-
tinue the experiment in the following years did not
allow the root systems to be destroyed for analysis.
We thus limited our analysis to the measurements of
diameters and depth.
The helophytes had root depths up to 40 cm, in particular
C. epigejos, P. australis and T. latifolia (Fig. 7). The latter’s
root system was also well developed in width, which is
particularly useful for producing a homogeneous filtering
system in floating wetlands. Conversely, halophytes devel-
oped smaller roots, reaching a maximum 15-cm depth, except
J. maritimus which reached a depth comparable to helophytes
roots (maximum 43 cm). The measurements refer to 2012
because it can be considered the first real growing
season in the system.
Conclusions
In conclusion, C. dactylon, T. latifolia, A. caerulescens,
E. atherica, H. portulacoides, P. palustris and S. maritima were
able to survive in the floating systems and in wastewater con-
centrations of the system with maximum values of 7.3 mS/cm
EC and of 225 mg/L NH4-N. Among these species, the highest
nitrogen uptakes were shown by P. palustris (14.93 g/m2 N in
2012), E. atherica (11.03 g/m2 N in 2012), S. maritima (10.40 g/
m2 N in 2011) and T. latifolia (7.94 g/m2 N in 2011).
This screening was aimed to investigate possible new
species to be used in floating systems and for the treatment
of digestate. Halophytes were promising for these uses, and
further testing and evaluation would be interesting.
Acknowledgments Research was carried out with the financial support
of the Italian Ministry of Agriculture and Forestry, FITOPROBIO Project.
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