Journal of Hazardous Materials 263 (2013) 486492

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Journal of Hazardous Materials

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

Removal of chromium ions from wastewater by duckweed,

Lemna minor L. by using a pilot system with continuous ow
Y. Uysal
Department of Environmental Engineering, Engineering and Architecture Faculty, Kahramanmaras Sutcu Imam University, Avsar Campus, Kahramanmaras
46100, Turkey

h i g h l i g h t s

Cr sorption potential of Lemna in different pH and concentration was determined.

Study was carried out in ponds to simulate wetlands and wastewater treatment pond.
Growth parameters of Lemna were investigated to determine toxic effects of Cr.
Increasing Cr concentration caused to increase Cr concentration.
Even plants suffered toxic effects; they continued to remove Cr from water.

a r t i c l e

i n f o

Article history:
Received 4 June 2013
Received in revised form 25 July 2013
Accepted 3 October 2013
Available online 11 October 2013
Keywords:
Chromium
Biosorption
Duckweed
Lemna minor
Wastewater

a b s t r a c t
The aim of this study was to determine the ability of Lemna minor to remove Cr (VI) ions from wastewater
in a continuous ow pond system. This system was used to simulate a wastewater treatment pond
and a natural wetland as habitat of plants. In order to nd optimal conditions for chromium removal,
ponds were operated with aqueous solutions having different pH (4.07.0) and chromium concentration
of 0.25 mgCr+6 /L, then plants were exposed to different chromium concentrations (0.255.0 mgCr+6 /L)
at pH 4.0. Chromium concentrations, both in biomass and wastewater, were measured and removal
efciency was determined throughout water ow. Growth factors such as growth rates, chlorophyll
contents and dry/fresh weight ratios of plants were also determined to measure toxic effects of chromium.
The percentages of chromium uptake (PMU) and bioconcentration factors (BCF) were calculated for each
run. The highest accumulated chromium concentration (4.423 mgCr/g) was found in plants grown in the
rst chamber of pond operated at pH 4.0 and 5.0 mgCr/L, while the minimum accumulated chromium
concentration (0.122 mgCr/g) was in plants grown in the last chamber of pond operated at pH 4.0 and
0.25 mgCr+6 /L.
2013 Elsevier B.V. All rights reserved.

1. Introduction
The main sources of heavy metal pollution in aquatic ecosystems are of the anthropogenic activities. Heavy metals include
arsenic, antimony, cadmium, chromium, mercury, lead are of particular concern to water and soil pollution. Chromium is one of
these heavy metals, and a widespread contaminant entering the air,
water and soil environment by different industrial activities such as
iron and steel manufacturing, chrome leather, chromium plating,
wood preservation and other anthropogenic sources. It exists in the
environment in two stable forms of chromium (III) and chromium
(VI) through natural processes and human activities.

Tel.: +90 344 280 1684; fax: +90 344 280 1602.
E-mail addresses: yuysal@ksu.edu.tr, yagmur uysal@hotmail.com
0304-3894/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.jhazmat.2013.10.006

The intensive use of chromium for several industrial applications causes high amount of wastewater containing chromium ions,
and also causes to increase its concentrations in the water sources
of environment. Chromium (VI) compounds are used in industry
for many purposes suh as: metal plating, cooling towers, leather
tanning, wood preservation. However, these compounds are highly
reactive, mobile and easily soluble in water, and these properties
cause several environmental health risks. Because, chromium (VI)
compounds are highly toxic to aquatic organisms and accumulated by their bodies. The problem of heavy metal contamination
consists in the fact that the metals cannot be degraded like other
organic xenobiotics but they must be extracted from a polluted
area [1]. Methods using living plants to remove metal ions from
a polluted area with organic and inorganic compounds have been
commonly called phytoremediation. Phytoremediation is a costeffective green technology based on the use of specially selected
metal-accumulating plants to remove toxic metal ions from soils

Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492

and water. Phytoremediation/constructed wetlands are claimed to

be low-technology systems able to treat a variety of wastewaters
[2] containing several heavy metal ions.
Aquatic vascular plants play an important role in the uptake,
storage, and recycling of metals, so many species of them are used
to remove these ions from wastewater. Live aquatic plant biomass
does not require any special preparations like dead biomass, and
it is typically grown in a separate chamber from the waste separation so that it can be physically separated from the wastewater
[3]. One of the most using aquatic families in the phytoremediation researches is Lemnaceae that is distributed globally in the
aquatic ecosystems. In general, these plants have been tested
and used for secondary or tertiary treatment (advanced), and
polishing of wastewater especially to remove metal ions and nutrients. Wastewater treatment with these plants provides effective,
reliable, and affordable solutions for pond-based municipal and
industrial treatment systems.
In the literature reports, duckweeds have also shown the ability
to accumulate chromium ions from water substantially. In addition,
duckweeds attain higher concentrations of chromium in their tissues than do other macrophytes. However, most of studies reported
in the literature were carried out using batch systems like asks
or Erlenmeyer. However, these oating plants were exposed to
continuous water ow both in nature and wastewater treatment
plants.
Wastewater is characterized by substantial variations in pH,
and may have different heavy metal ions at variety concentration
range. In addition, treatment processes mostly occur in natural
systems depending on the metabolic activities of plants, and pH
value of water that they possibly affect the growth of plants.
Thus, choosing of optimal pH value for the metabolism-dependent
removal and bioaccumulation of pollutants is quietly important
operation parameter in order to achieve requested treatment level.
Lemna is a fast growing plant species and adapts easily to various aquatic conditions like pH, temperature and light. Plants are
able to grow across a wide range of pH (3.510.5). Although this
wide pH range, the optimal pH range for growth is 4.57.5 [4]. The
plant carries on growing, but growth rate decreases at the lower
or upper limits of this pH range. Thus, this study was carried out
to determine the effect of different wastewater conditions such
as pH (4.07.0) and chromium concentration (0.255.0 mgCr/L)
on the sorption by Lemna minor. In order to simulate a stabilization pond for wastewater treatment and also simulate a natural
wetland for plants habitat, a laboratory scale pilot system with
continuous water ow composed of three ponds was used in this
study. The amount of plant biomass produced was also determined during the study period on weight and dry/fresh weight ratio
basis.

487

Fig. 1. Pilot system with L. minor.

They were rst washed carefully with tap water several times to
remove dirt, sludge and other debris, then were set into pilot system
consisted of three plexiglas ponds with a capacity of 14 L lled with
Synthetic Duckweed Nutrient Solution (DNS) [7]. No detectable
levels of chromium were found in the water or sediments at the
collection site.

2.3. Pilot system

This system consisted of three ponds as shown in Fig. 1. All
the experiments were carried out in two ponds. One pond was
used as control pond without any addition of metal. They were
fed with DNS using a peristaltic pump continuously. Temperature
was maintained at 2325 C, and 16 h of photoperiod was applied
to plants at a photosynthetic photon density of 250 mol/m2 s at
plant level. Each pond was segmented into twelve 10 10 cm sections connected to each other, and hydraulic retention times (HRT)
for the ponds were chosen as 7 days. Water samples were collected from the efuent of chromium pond for chromium analyses
periodically.

2.4. Growth parameters

2. Experimental
2.1. Terminology
In this study, differentiation between chromium oxidation
states is not a concern; similarly to other works [5,6], biouptake
including active incorporation into living cells as well as physical
adsorption will be referred to total Cr. Two separate experiments
were conducted: (1) to determine chromium removal from water
by duckweed and (2) to determine the inuence of Cr on duckweed
growth and its biochemical parameters.
2.2. Experimental plant material
Plants were collected from freshwater canals in Kahramanmaras, and transferred to the climate chamber in the laboratory.

In order to determine any possible toxic effects of chromium

on plants, biochemical parameters such as growth rates, dry/fresh
weight ratios and chlorophyll contents of plants were determined.
Duckweed growth was determined by measuring fresh weight
(FW, biomass) according to the Standard Methods [7]. Plants were
surface-dried between layers of paper towels, and fresh weight
was determined. To measure dry weight, plants were dried at
80 C up to constant weight (usually 24 h). Since experiments were
carried out in the ponds, and amount of plants was too high to
count of frond number, special rings were used to limit the area
of plants. These rings were set into chambers of ponds of 1st, 3th,
5th, 8th, 10th and 12th to calculate growth rates. The inner space
areas of rings were about 20 cm2 and plants (1 g) were put into
them at the beginning of each experiment. Typical plant size was
two to four fronds on each plant; and plant density was sufcient
to exclude any signicant algae or other plant growth.

488

Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492

2.5. Test procedures

100
pH 4
pH 5
pH 6
pH 7

80
Percentage removal of Cr +6

The nutrient medium of DNS was prepared by using the following salts in stock solution: NaNO3 , 25.50 g/L; NaHCO3 , 15.0 g/L;
K2 HPO4 , 1.04 g/L; CaCl2 2H2 O, 4.41 g/L; MgCl2 , 5.70 g/L; FeCl3 ,
0.096 g/L; MnCl2 , 0.26 g/L; MgSO4 .7H2 O, 14.7 g/L; H3 BO3 , 0.186 g/L;
Na2 MoO4 2H2 O, 7.26 mg/L; ZnCl2 , 3.27 mg/L; CoCl2 , 0.78 mg/L; and
CuCl2 , 0.009 mg/L. This stock solution was diluted with deionized
water at the ratios of 1:100 to make the plant nutrient medium.
Chromium solutions at nominal concentrations of 0.25, 0.5, 1.0,
2.0, 4.0 and 5.0 mg/L were added directly to the DNS. The initial
pH of solutions was adjusted by using HCI and NaOH solutions,
and the change of pH was monitored daily. In order to determine
the chromium accumulation in plant body, plants were collected
from the chambers of chromium pond by ltering, and were then
washed with deionized water. Then they were dried to determine dry weights (Xm). In order to analyze bioaccumulated metal
concentrations, dried plants were digested with HNO3 and H2 SO4
mixture solution according to the process of Sen and Mondal [8],
and diluted to certain volume to measure the accumulated metal
ion per unit mass (mgCr/g). The concentrations of chromium in
all digested water and plant samples were determined by inductively coupled plasma optical emission spectrometry (ICP-OES,
Optima 2100-Perkin Elmer). A reagent blank was also digested in
the same manner. Chlorophyll contents of plant tissues were also
determined. The chlorophyll a (chl-a) and b (chl-b) levels were
determined on a fresh weight basis according to Standard Methods
[7].

60

40

20

0
0

10

12

14

16

Number of Days

Fig. 2. Percentage removal of Cr from feed solutions having different pH values

(pH = 4.07.0, Cr+6 = 0.25 mg/L).

growth rate (RGR) (g day1 ) was calculated using the following

equation [3] by Greger et al. [10].
RGR =

dW
(ln Wt2 ln Wt1 )
=
t2 t1
dt

(3)

RGR: growth rate (fresh weight/d); Wt1 : the fresh weight at t1 ; Wt2 :
the fresh weight at t2 .

2.6. Reagents
All reagents used were of analytical grade. Chromium stock solutions were prepared with potassium dichromate (K2 Cr2 O7 ) (Merck)
at nominal concentrations of 0.25, 0.5, 1.0, 2.0, 4.0 and 5.0 mg/L by
pipetting appropriate amounts of 1000 mg/L stock solution. Nitric
acid and sulphuric acid were metal-analysis grade (Merck, German). Dilute nitric acid (2% by weight) was used for cleaning and
rinsing of all equipment and sampling materials.

2.7. Calculations
The bioremoval potential of Lemna was expressed as percentage
of metal ion uptake (PMU) and calculated according to following Eq.
(1).
PMU (the percentage of Cr uptake) =
Mass of Cr in Lemna
100
Mass of Cr in medium(t = 0)

(1)

Bioconcentration of a chemical by aquatic organisms is generally expressed as the bioconcentration factor (BCF). This factor is an
indicator of the metal accumulation ability of plants relative to the
metal concentration in medium (Eq. (2)), and provides the comparison of the results between species. BCF values of L. minor for
studied conditions were calculated according to Zayed et al. [9].
BCF =

mg chromium/kg Lemna
mg chromium/L solution

(2)

Commonly, plant growth was explained depending on the

increase of the frond number of Lemna. However, our experiments
were carried out in ponds and amount of plants were too high to
count the frond number, so we calculated growth rates as fresh
weight increase of plants in certain time. The average relative

3. Results and discussion

3.1. Bioaccumulation of chromium at different pH values by L.
minor
In order to determine the effect of different initial pH values
of culture medium on chromium accumulation and plant growth,
ponds were fed with DNS having Cr (VI) concentration of 0.25 mg/L
at different pH values (4.07.0). A control pond was also fed with
DNS having same pH value and no metal. Each experiment was
run for 7 days. Samples were taken the rst day at 0 h (before
the addition of biomass), and at certain time interval to measure
residual chromium concentrations in the efuent. The percentage
chromium removal values are shown in Fig. 2.
It can be seen from Fig. 2 that percentage chromium removal
efciency of system continuously decreased with time. Chromium
removal for all pH values is in the range 88.834.0% for rst 7
days of experimental period, and this range decreased to range
76.420.0% for 714 days. However, it can be said from the results
that the actual removal efciency of Lemna system started after
7th day of experimental period, because HRT of ponds were 7 day.
A slightly decreased was observed in the efuent Cr concentration
(0.855 mg/L for 1.0 mgCr+6 /L initial concentration) of control pond
on 14th day.
The decreasing removal rate by aquatic plants with increasing
contact time is indicative of a fast attainment of saturation state in
the plants [11]. Similar results have been reported by our earlier
study [12] for the removal of Pb(II) using L. minor. It was found
that Pb(II) concentrations in water did not change with initial
pH, and declined rapidly at rst 24 h, then slowly to 72 h, and
remained stable after 72 h. Maine et al. [13] also reported that Cr
was efciently depleted from water, and the greatest decrease
was observed during the rst 24 h of the experiments about
7083% and 5880% of the added Cr concentration (1.02.0 mg/L)
for Salvinia herzogii and Pistia stratiotes, respectively. Chromium

Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492

489

Table 1
PMU% and BCF values of Lemna at different pH values (pH 47; Cr+6 = 0.25 mg/L).
Chambers of pond

pH
4.0

1
3
5
8
10
12

5.0

6.0

BCF

PMU%

BCF

PMU%

BCF

PMU%

BCF

PMU%

1996
1123
932
1112
1072
488

16.4
11.8
17.4
57.2
50.8
5.7

776
498
408
352
304
284

4.0
3.2
2.8
3.1
2.5
2.2

688
580
468
448
388
296

8.7
7.6
5.3
6.6
5.7
2.3

336
376
436
360
332
380

2.5
2.6
2.5
2.4
2.3
2.4

0,6
pH 4
pH 5
pH 6
pH 7

0,5

Cr (mg/g)

0,4

0,3

0,2

0,1

0,0
0

7.0

10

12

Chambers of chromium pond

Fig. 3. Effect of culture pH on the Cr bioaccumulation by L. minor (Cr+6 = 0.25 mg/L).

concentrations accumulated in the plant biomass after 14 days of

contact time are shown in Fig. 3.
Fig. 3 shows that maximum accumulation (0.499 mgCr/g) was
obtained at pH 4.0. Plants grown in the initial chambers of pond
(1st and 3th) where they exposed to chromium ions rstly, accumulated more chromium ions than plants grown in the other
chambers of pond (5th, 8th, 10th and 12th). Thus, decreasing
uptake of chromium ions resulted from decreasing concentration
of chromium ions in the water. PMU(%) and BCF values were also
calculated and are shown in Table 1.
The maximum PMU value was obtained in the 8th chamber of
chromium pond operated at pH 4.0 as 57.2% while, the lowest PMU
value was obtained as 2.2% in the last chamber of pond operated at
pH 5.0. However, it was observed that PMU values obtained at pH
7.0 and 5.0 were the lowest. The ndings showed that accumulated
chromium concentrations decreased with increasing of pH values
of medium higher than 4.0. The highest growth rates of plants were
obtained in the 1011th chambers of chromium pond, while plants
were smaller and in light color in the 1st chamber of pond. Similarly,
the highest BCF value was obtained in the 1st chamber of pond at pH
4.0 as 1996. Presence of chromium ions in the aquatic environment
of plants leads to changes in their growth and development. In this
work, toxic effects of chromium were investigated as changes of
some metabolic parameters such as growth rates, dry/fresh weight
ratios and reduction of chlorophyll contents (Fig. 4ad).
Fig. 4(ad) shows that maximum growth rate values were
obtained at pH values of 4.0 and 6.0. Generally, growth rates of
plants showed an increase between chambers of 1st10th, and then
they showed a decrease in the last chamber of chromium pond.
Maximum growth rates were generally obtained in the 5th chamber of chromium pond and 10th chamber of control pond at pH
4.0. Plants were small and in light green in the rst chamber of

chromium pond but they were healthier up to pond end. Similarly,

in the control pond, plants grown in the chambers of 6th11th were
healthier than others. There was an interest nding in the observations that the healthiest plants in bright color were obtained at
pH 4.0 similarly growth rate results. It is suggested that chromium
concentration of 0.25 mg/L induced the plant growth at this pH
value. However, D/F ratios of plants growing in the rst chamber of pond were similar to those of controls, but they showed a
rapid increase between chambers of 1st and 10th. While D/F ratios
for chromium pond were obtained in the range 0.0500.245 d1 ,
this range was obtained as only 0.0390.114 d1 for control. The
increase of dry mass compared to control plants is possibly related
with high chromium accumulation at pH 4.0. Another reason of
this may be starch storing of fronds. Hillman [15] detected that
duckweed fronds can store starch under the stress conditions. Similarly, Dirilgen and Inel [16] reported that dry biomass amount of
L. minor increased due to storing of starch when exposed to toxic
metal ions and stress conditions. Because the growth rates of plants
showed similar trend with control, this increase in the dry masses
may be the indicator of stress conditions caused by chromium
ions.
Growth rates and D/F ratios values were about same for control
and chromium ponds at pH 5.0. While growth rates were in the
range 0.0150.066 d1 for chromium pond, they were obtained as
0.0140.075 d1 for control pond. Growth rates of plants grown in
the chromium pond were also similar to those of control plants at
pH 7.0. However, D/F ratios of plants grown in the chromium pond
were generally slightly higher than those of control plants.
Chromium stress is one of the important factors affecting photosynthesis in terms of CO2 xation, electron transport,
photophosphorylation and enzyme activities [17]. Landolt and
Kandeler [18] reported that chromium affects especially the photosystem II for Lemna aequinoctialis. The inuence of 1 and 2 mg/L
Cr(VI) on Salvinia minima decreased chlorophylls a and b and
carotenoid concentrations signicantly [19]. In the present study,
results obtained from the chlorophyll analyses were summarized in
Fig. 5. Chlorophyll contents of plants decreased according to those
of control plants signicantly for pH 4.0 at which accumulation was
highest. The reason of this decrease could probably resulted from
stress effects of chromium accumulation.
3.2. Bioaccumulation of chromium at increasing chromium (VI)
concentration of medium by L. minor
In order to determine different chromium concentration in the
medium on the bioaccumulation efciency of Lemna, chromium
concentrations of inuent were increased to 0.5, 1.0, 2.0, 4.0
and 5.0 mg/L, and pH adjusted to 4.0. Calculated percentages of
chromium removal values for each concentration are shown in
Fig. 6.
Similarly, the percentage removal of chromium continuously
decreased with time. Cr was efciently depleted from water and
the greatest decrease was observed during the rst 24 h of the

490

Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492

0,25
CHAMBERS vs GROWTH
CHAMBERS vs D/F RATIO
GROWTH-C
D/F RATIO-C

0,20

0,08

CHAMBERS vs GROWTH
CHAMBERS vs D/F RATIO
GROWTH-C
D/F-C

pH 4

pH 5

0,06

0,15

0,04
0,10

0,02
0,05

0,00

0,00
0

10

12

6
8
Chambers of ponds

Chambers of ponds

0,08

d 0,08

CHAMBERS vs GROWTH
CHAMBERS vs DRY/FRESH RATIO
GROWTH -C
D/F RATIO-C

0,06

0,04

0,04

0,02

0,02

12

CHAMBERS vs GROWTH
CHAMBERS vs DRY/FRESH RATIO
GROWTH-C
DRY/FRESH RATIO-C

pH 6

0,06

10

pH 7

0,00

0,00
0

10

12

10

12

Chambers of ponds

Chambers of ponds

Fig. 4. (a)(d) Effect of culture pH on the growth of Lemna minor grown in the different chambers of ponds.

experiments (about 9965% of the added Cr for L. minor). Chromium

removal efciency is in the range 6641% for rst 7 days (HRT rst),
and this range decreased to range 4823% for 714 days (HRT second) while, it was again decreased to the range 4922% for 1421
days (HRT third). During the experimental period (21 days), L. minor
removed approximately 40.8%, 48.5%, 29.0%, 26.0% and 21.8% of the
added chromium ions in the concentrations of 0.5, 1.0, 2.0, 4.0 and
5.0 mg/L, respectively.

Effect of increasing chromium concentration on the growth

parameters (RGR, dry weight, dry/fresh weight ratio) of Lemna
were summarized in Table 2. The change of these parameters of
plants by chromium exposure was shown with ponds chambers
(water ow). According to the results, it can be seen that Cr concentration in the water inuenced the average relative growth rate
(RGR) of plants presenting a signicant negative linear correlation. Dry weights of plants increased between the chambers of
rst to 10 in the concentrations of 0.5 and 1.0 mgCr/L. This increase

180

Chlorophyll (control=100%)

140
120
100
80
60
40

100
90
80

Percentage Removal of Cr+6

pH 4
pH 5
pH 6
pH 7
Control

160

70
60
50
40
30
20
10
0

20

10

15

20

25

Time (days)
0

10

12

0.5 ppm

1 ppm

2 ppm

4 ppm

5 ppm

Chambers of ponds

Fig. 5. Chlorophyll content of Lemna minor at different pH 4.07.0 (Cr+6 = 0.25 mg/L).

Fig. 6. Percentage removal of chromium from feed solutions having different Cr (VI)
concentration (pH = 4.0; Cr+6 = 0.55.0 mg/L).

Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492

Table 2
Growth parameters of Lemna with increasing chromium concentration of culture
medium (pH 4; Cr+6 = 0.25-5.0 mg/L).
Cr (mg/L)

Chambers of chromium pond

1

10

12

0.25
Dry weight (g)
RGR
D/F

0.101
0.035
0.062

0.105
0.033
0.067

0.187
0.070
0.070

0.515
0.067
0.203

0.474
0.050
0.235

0.117
0.038
0.069

0.5
Dry weight (g)
RGR
D/F

0.051
0.001
0.050

0.115
0.055
0.045

0.126
0.040
0.064

0.145
0.063
0.050

0.149
0.050
0.064

0.121
0.047
0.055

1.0
Dry weight (g)
RGR
D/F

0.038
0.005
0.042

0.147
0.054
0.047

0.136
0.031
0.071

0.176
0.047
0.066

0.183
0.031
0.095

0.132
0.047
0.050

2.0
Dry weight (g)
RGR
D/F

0.108
0.020
0.082

0.125
0.021
0.088

0.128
0.023
0.087

0.150
0.035
0.082

0.118
0.013
0.094

0.106
0.014
0.083

4.0
Dry weight (g)
RGR
D/F

0.108
0.016
0.135

0.125
0.023
0.085

0.128
0.005
0.139

0.150
0.008
0.132

0.118
0.001
0.120

0.106
0.019
0.148

5.0
Dry weight (g)
RGR
D/F

0.033
0.066
0.102

0.029
0.055
0.074

0.026
0.077
0.096

0.021
0.067
0.065

0.026
0.062
0.075

0.019
0.122
0.150

was observed between the chambers of rst to 8. However, in the

5.0 mgCr/L, dry weights of plants showed slightly decrease with
ponds chambers due to toxic effect of chromium. Bioaccumulated
chromium concentrations, BCF, PMU% and relative chlorophyll content values for different chromium concentrations were shown in
Table 3.
An increase in the amount of accumulated chromium concentrations by plant biomass was observed with increasing metal
concentration of medium (Table 3). In the chromium solutions of
4.0 and 5.0 mgCr/L, L. minor was able to accumulate chromium ions
in its body up to concentrations of 1.611 and 4.423 mg/g, respectively. These values are quite higher than those of other plants
reported in the literature such as water lily, Nymphaea spontanea
as 1.466 mg/g [20], and Bacopa monnieri as 1.600 mg/g [21] when
exposed to same chromium concentration. According to the results
summarized in Table 3, the highest BCF value was obtained in
the lowest Cr supply (0.25 mg/L) as 1996. BCF values decreased
when Cr supply was increased from 0.25 to 2.0 mg/L and with
water ow through the pond. The lowest BCF value was obtained
in the last chamber of pond having 2.0 mg/L inuent chromium
concentration as 149, and these values showed an increase again
for inuent chromium concentrations higher than 2.0 mg/L. It can
be seen from the results that BCF values are higher for chromium
concentrations of 4.0 and 5.0 mg/L than those of 2 mg/L (Table 3).
However, decreasing of chlorophyll contents of plants in these
high chromium concentrations showed that this increase was not
resulted from biouptake or biochemical metabolism by plants.
When BCF values are investigated in the literature, it can be seen
that these values are higher in the studies performed at very low
metal ion concentrations than those in performed with moderate
and high metal ion concentrations. Arora et al. [22] investigated
the potential of Azolla spp. namely A. microphylla, A. pinnata and A.
liculoides to tolerate chromium ions in their growth environment
and bioconcentrate them. They reported that BCF values were in
the range 2434617 for three species. A. microphylla showed highest bioconcentration potential, while its growth was affected to

491

Table 3
Change of chromium accumulation (mg/g), BCF, PMU%, and chlorophyll content
of Lemna with increasing chromium concentration of culture medium (pH 4;
Cr+6 = 0.255.0 mg/L).
Cr (mg/L)

Chambers of chromium pond

1

10

12

0.2
Cr (mg/g)
BCF
PMU%
Chlorophyll (control %)

0.499
1996
16.4
41.69

0.281
1123
11.8
29.95

0.233
932
17.4
51.30

0.278
1112
57.2
51.54

0.268
1072
50.8
37.79

0.122
488
5.7
51.80

0.5
Cr (mg/g)
BCF
PMU%
Chlorophyll (control %)

0.484
968
4.9
42.48

0.335
670
7.7
46.94

0.375
750
9.5
49.22

0.232
464
6.7
51.57

0.227
454
6.7
58.07

0.160
320
3.9
46.55

1.0
Cr (mg/g)
BCF
PMU%
Chlorophyll (control %)

1.038
1038
22.5
48.11

0.426
426
5.5
48.46

0.487
487
7.3
47.71

0.238
238
6.6
43.26

0.326
326
8.6
55.49

0.311
311
9.1
57.18

2.0
Cr (mg/g)
BCF
PMU%
Chlorophyll (control %)

0.634
317
3.4
9.16

0.616
308
3.9
14.00

0.392
196
2.5
17.10

0.353
177
2.6
21.68

0.343
172
2.0
27.00

0.298
149
1.6
30.31

4.0
Cr (mg/g)
BCF
PMU%
Chlorophyll (control %)

1.611
403
1.6
16.51

0.995
249
1.9
19.68

1.203
301
1.7
13.24

0.908
227
1.6
22.38

0.647
162
0.9
13.04

0.958
240
0.9
29.17

5.0
Cr (mg/g)
BCF
PMU%
Chlorophyll (control %)

4.423
885
2.9
1.62

3.782
756
2.2
1.43

4.412
882
2.3
1.40

3.459
692
1.4
1.77

4.099
820
2.1
2.00

3.544
709
1.3
2.11

greater extent than those of A. pinnata and A. liculoides. Landolt and

Kandeler [18] listed BCF values for different species of Lemnaceae
in their book (which name is The Family of Lemnaceae a monographic study), and showed them to be in the range 38040000 for
chromium. In the present study, BCF values for different chromium
concentrations were obtained in the range 1491996, and this
range shows that L. minor appears to be a poor accumulator for
chromium.
When considering relative chlorophyll contents of plants as the
percent of control, it was shown that total chlorophyll amounts
were not affected by increasing chromium concentration up to
1.0 mg/L. Chlorophyll contents of plants was average 50% of control
plants in the 0.1 mg/L chromium concentration while, it decreased
to about 20% for chromium concentration of 2.0 mg/L. However,
this value signicantly decreased to 1.8% for 5 mg/L chromium concentration, because all plants were whitened and died in the rst
24 h of this run. Similar results were also reported in the literature
that chlorophyll content of P. stratiotes decreased with increasing metal concentration upper than 1.0 mg/L in medium [13].
Decreasing of chlorophyll content of plants suggests that metals
affected the chlorophyll synthesizing system and chlorophyllase
activity. A decrease in the photosynthetic pigment content is well
known from many other works concerning chromium stress in
plants and it is attributed to the oxidative stress [21]. Augustynowicz et al. [1] reported that very strong declining of photosynthetic
activity was observed when plants exposed to chromium ions. They
reported that 100 M Cr was the border concentration at which
some symptoms of the metal toxicity on the C. cophocarpa (waterstarwort) were found. They observed inhibition of growth and
decrease in the photosynthetic pigment content at this concentration. Shanker et al. [14] reported that disorganization of chloroplast

492

Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492

ultrastructure and inhibition of electron transport processes due to

chromium ions is a possible explanation for Cr-induced decrease in
photosynthetic rate of plant cells.
In the present study, author tried chromium concentration at
the range 0.5100 M, and toxicity symptoms were observed in
the chromium concentration higher than 2 M. However, even
plants may have been suffering toxic effects due to the bioaccumulation of chromium ions; they continued to remove them
from water. Chlorophyll contents and accumulation efciencies of
plants decreased with increasing chromium concentration while,
BCF values showed an increase higher than 2 mg/L chromium concentration in which toxicity symptoms appeared.
4. Conclusions
The capacity of aquatic plants to remove potentially toxic heavy
metals, including chromium, from water is well documented. When
considering the literature studies, it can be seen that much studies were conducted in batch systems and asks. The difference of
this study is carrying out in a pilot system with continuous water
ow in order to simulate a wastewater treatment pond. The results
of this study provide information about the dependence of biouptake capacity of L. minor on operational pH value and chromium
concentration of wastewater. It was proved that pilot system containing of ponds with continuous water ow like a natural and/or
constructed wetland could efciently reduce chromium contents
of water. However, chromium ions showed some toxic effects on L.
minor upper than concentration of 2.0 mg/L apparently, but system
continued to remove chromium ions with low efciency. Consequently, wastewater containing a low chromium concentration can
be treated using these plants by continuously subjecting them to
water ow. In addition to this, in order to prevent the release of
accumulated metal ions from plant biomass, Lemnaceae cover must
be removed by harvesting of plants regularly in certain time.
Conict of interest
No competing nancial interests exist.
Acknowledgements
This work was a part of the project No: 2011/3-49M supported
by the Unit of Research Project (BAP) of Kahramanmaras Sutcu
Imam University. The author is grateful to KSU-BAP for the nancial
support provided for the pursuit of this project.

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