Journal of Hazardous Materials 263 (2013) 486492

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Journal of Hazardous Materials

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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 a b s t r a c t

Article history: The aim of this study was to determine the ability of Lemna minor to remove Cr (VI) ions from wastewater
Received 4 June 2013 in a continuous ow pond system. This system was used to simulate a wastewater treatment pond
Received in revised form 25 July 2013 and a natural wetland as habitat of plants. In order to nd optimal conditions for chromium removal,
Accepted 3 October 2013
ponds were operated with aqueous solutions having different pH (4.07.0) and chromium concentration
Available online 11 October 2013
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
Keywords:
efciency was determined throughout water ow. Growth factors such as growth rates, chlorophyll
Chromium
Biosorption
contents and dry/fresh weight ratios of plants were also determined to measure toxic effects of chromium.
Duckweed The percentages of chromium uptake (PMU) and bioconcentration factors (BCF) were calculated for each
Lemna minor run. The highest accumulated chromium concentration (4.423 mgCr/g) was found in plants grown in the
Wastewater 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 intensive use of chromium for several industrial applica-

The main sources of heavy metal pollution in aquatic ecosys-
tems are of the anthropogenic activities. Heavy metals include
arsenic, antimony, cadmium, chromium, mercury, lead are of par-
ticular 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
tions 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 accumu-
lated 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 cost-
effective green technology based on the use of specially selected
metal-accumulating plants to remove toxic metal ions from soils

0304-3894/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.jhazmat.2013.10.006
 Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492 487

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 sepa-
ration so that it can be physically separated from the wastewater
[3]. One of the most using aquatic families in the phytoremedia-
tion 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 nutri-
ents. 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, Fig. 1. Pilot system with L. minor.

duckweeds attain higher concentrations of chromium in their tis-

sues 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 vari-
ous 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 stabiliza-
tion 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 deter-
mined during the study period on weight and dry/fresh weight ratio
basis.
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 sec-
tions connected to each other, and hydraulic retention times (HRT)
for the ponds were chosen as 7 days. Water samples were col-
lected 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 Kahraman-
maras, 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
The nutrient medium of DNS was prepared by using the fol-
lowing 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 deter-
mine 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 induc-
tively 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].
2.6. Reagents
All reagents used were of analytical grade. Chromium stock solu-
tions 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, Ger-
man). 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)
Bioconcentration of a chemical by aquatic organisms is gener-
ally 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 com-
parison of the results between species. BCF values of L. minor for
studied conditions were calculated according to Zayed et al. [9].
mg chromium/kg Lemna
BCF =
mg chromium/L solution
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
100
pH 4
pH 5
pH 6
80
pH 7
Percentage removal of Cr +6
60
40
20
0
0 2 4 6 8 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].
dW (ln Wt2 ln Wt1 )
RGR = = (3)
dt t2 t1
RGR: growth rate (fresh weight/d); Wt1 : the fresh weight at t1 ; Wt2 :
the fresh weight at t2 .
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
(1)
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
(2) 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 5.0 6.0 7.0

BCF PMU% BCF PMU% BCF PMU% BCF PMU%

1 1996 16.4 776 4.0 688 8.7 336 2.5

3 1123 11.8 498 3.2 580 7.6 376 2.6
5 932 17.4 408 2.8 468 5.3 436 2.5
8 1112 57.2 352 3.1 448 6.6 360 2.4
10 1072 50.8 304 2.5 388 5.7 332 2.3
12 488 5.7 284 2.2 296 2.3 380 2.4

0,6 chromium pond but they were healthier up to pond end. Similarly,
pH 4
in the control pond, plants grown in the chambers of 6th11th were
pH 5 healthier than others. There was an interest nding in the obser-
0,5
pH 6 vations that the healthiest plants in bright color were obtained at
pH 7
pH 4.0 similarly growth rate results. It is suggested that chromium
0,4 concentration of 0.25 mg/L induced the plant growth at this pH
value. However, D/F ratios of plants growing in the rst cham-
Cr (mg/g)

ber of pond were similar to those of controls, but they showed a

0,3
0,2
0,1
0,0
0 2 4 6 8 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, accu-
mulated 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 cham-
ber 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
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. Sim-
ilarly, 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 affect-
ing photosynthesis in terms of CO2 xation, electron transport,
photophosphorylation and enzyme activities [17]. Landolt and
Kandeler [18] reported that chromium affects especially the pho-
tosystem 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

a 0,25 b 0,08
CHAMBERS vs GROWTH
CHAMBERS vs GROWTH CHAMBERS vs D/F RATIO
CHAMBERS vs D/F RATIO pH 4 pH 5
GROWTH-C
GROWTH-C D/F-C
0,20 D/F RATIO-C
0,06

0,15

0,04

0,10

0,02
0,05

0,00 0,00
0 2 4 6 8 10 12 0 2 4 6 8 10 12
Chambers of ponds Chambers of ponds

c 0,08
CHAMBERS vs GROWTH d 0,08 CHAMBERS vs GROWTH
CHAMBERS vs DRY/FRESH RATIO CHAMBERS vs DRY/FRESH RATIO
pH 6 pH 7
GROWTH -C GROWTH-C
D/F RATIO-C DRY/FRESH RATIO-C

0,06 0,06

0,04 0,04

0,02 0,02

0,00
0,00
0 2 4 6 8 10 12
0 2 4 6 8 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 sec-
ond) 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 con-
centration in the water inuenced the average relative growth rate
(RGR) of plants presenting a signicant negative linear correla-
tion. 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
pH 4 100
160 pH 5 90
pH 6
140 pH 7 80
Percentage Removal of Cr+6
Chlorophyll (control=100%)

Control
70
120
60
100
50
80 40
30
60
20
40
10
20 0
0 5 10 15 20 25
0 Time (days)
0 2 4 6 8 10 12 0.5 ppm 1 ppm 2 ppm 4 ppm 5 ppm
Chambers of ponds
Fig. 6. Percentage removal of chromium from feed solutions having different Cr (VI)
Fig. 5. Chlorophyll content of Lemna minor at different pH 4.07.0 (Cr+6 = 0.25 mg/L). concentration (pH = 4.0; Cr+6 = 0.55.0 mg/L).
 Y. Uysal / Journal of Hazardous Materials 263 (2013) 486492 491

Table 2 Table 3
Growth parameters of Lemna with increasing chromium concentration of culture Change of chromium accumulation (mg/g), BCF, PMU%, and chlorophyll content
medium (pH 4; Cr+6 = 0.25-5.0 mg/L). 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
Cr (mg/L) Chambers of chromium pond
1 3 5 8 10 12
1 3 5 8 10 12
0.25
Dry weight (g) 0.101 0.105 0.187 0.515 0.474 0.117 0.2
RGR 0.035 0.033 0.070 0.067 0.050 0.038 Cr (mg/g) 0.499 0.281 0.233 0.278 0.268 0.122
D/F 0.062 0.067 0.070 0.203 0.235 0.069 BCF 1996 1123 932 1112 1072 488
PMU% 16.4 11.8 17.4 57.2 50.8 5.7
0.5
Chlorophyll (control %) 41.69 29.95 51.30 51.54 37.79 51.80
Dry weight (g) 0.051 0.115 0.126 0.145 0.149 0.121
RGR 0.001 0.055 0.040 0.063 0.050 0.047 0.5
D/F 0.050 0.045 0.064 0.050 0.064 0.055 Cr (mg/g) 0.484 0.335 0.375 0.232 0.227 0.160
BCF 968 670 750 464 454 320
1.0
PMU% 4.9 7.7 9.5 6.7 6.7 3.9
Dry weight (g) 0.038 0.147 0.136 0.176 0.183 0.132
Chlorophyll (control %) 42.48 46.94 49.22 51.57 58.07 46.55
RGR 0.005 0.054 0.031 0.047 0.031 0.047
D/F 0.042 0.047 0.071 0.066 0.095 0.050 1.0
Cr (mg/g) 1.038 0.426 0.487 0.238 0.326 0.311
2.0
BCF 1038 426 487 238 326 311
Dry weight (g) 0.108 0.125 0.128 0.150 0.118 0.106
PMU% 22.5 5.5 7.3 6.6 8.6 9.1
RGR 0.020 0.021 0.023 0.035 0.013 0.014
Chlorophyll (control %) 48.11 48.46 47.71 43.26 55.49 57.18
D/F 0.082 0.088 0.087 0.082 0.094 0.083
2.0
4.0
Cr (mg/g) 0.634 0.616 0.392 0.353 0.343 0.298
Dry weight (g) 0.108 0.125 0.128 0.150 0.118 0.106
BCF 317 308 196 177 172 149
RGR 0.016 0.023 0.005 0.008 0.001 0.019
PMU% 3.4 3.9 2.5 2.6 2.0 1.6
D/F 0.135 0.085 0.139 0.132 0.120 0.148
Chlorophyll (control %) 9.16 14.00 17.10 21.68 27.00 30.31
5.0
4.0
Dry weight (g) 0.033 0.029 0.026 0.021 0.026 0.019
Cr (mg/g) 1.611 0.995 1.203 0.908 0.647 0.958
RGR 0.066 0.055 0.077 0.067 0.062 0.122
BCF 403 249 301 227 162 240
D/F 0.102 0.074 0.096 0.065 0.075 0.150
PMU% 1.6 1.9 1.7 1.6 0.9 0.9
Chlorophyll (control %) 16.51 19.68 13.24 22.38 13.04 29.17

5.0
was observed between the chambers of rst to 8. However, in the Cr (mg/g) 4.423 3.782 4.412 3.459 4.099 3.544
5.0 mgCr/L, dry weights of plants showed slightly decrease with BCF 885 756 882 692 820 709
PMU% 2.9 2.2 2.3 1.4 2.1 1.3
ponds chambers due to toxic effect of chromium. Bioaccumulated
Chlorophyll (control %) 1.62 1.43 1.40 1.77 2.00 2.11
chromium concentrations, BCF, PMU% and relative chlorophyll con-
tent values for different chromium concentrations were shown in
Table 3. greater extent than those of A. pinnata and A. liculoides. Landolt and
An increase in the amount of accumulated chromium concen- Kandeler [18] listed BCF values for different species of Lemnaceae
trations by plant biomass was observed with increasing metal in their book (which name is The Family of Lemnaceae a mono-
concentration of medium (Table 3). In the chromium solutions of graphic study), and showed them to be in the range 38040000 for
4.0 and 5.0 mgCr/L, L. minor was able to accumulate chromium ions chromium. In the present study, BCF values for different chromium
in its body up to concentrations of 1.611 and 4.423 mg/g, respec- concentrations were obtained in the range 1491996, and this
tively. These values are quite higher than those of other plants range shows that L. minor appears to be a poor accumulator for
reported in the literature such as water lily, Nymphaea spontanea chromium.
as 1.466 mg/g [20], and Bacopa monnieri as 1.600 mg/g [21] when When considering relative chlorophyll contents of plants as the
exposed to same chromium concentration. According to the results percent of control, it was shown that total chlorophyll amounts
summarized in Table 3, the highest BCF value was obtained in were not affected by increasing chromium concentration up to
the lowest Cr supply (0.25 mg/L) as 1996. BCF values decreased 1.0 mg/L. Chlorophyll contents of plants was average 50% of control
when Cr supply was increased from 0.25 to 2.0 mg/L and with plants in the 0.1 mg/L chromium concentration while, it decreased
water ow through the pond. The lowest BCF value was obtained to about 20% for chromium concentration of 2.0 mg/L. However,
in the last chamber of pond having 2.0 mg/L inuent chromium this value signicantly decreased to 1.8% for 5 mg/L chromium con-
concentration as 149, and these values showed an increase again centration, because all plants were whitened and died in the rst
for inuent chromium concentrations higher than 2.0 mg/L. It can 24 h of this run. Similar results were also reported in the literature
be seen from the results that BCF values are higher for chromium that chlorophyll content of P. stratiotes decreased with increas-
concentrations of 4.0 and 5.0 mg/L than those of 2 mg/L (Table 3). ing metal concentration upper than 1.0 mg/L in medium [13].
However, decreasing of chlorophyll contents of plants in these Decreasing of chlorophyll content of plants suggests that metals
high chromium concentrations showed that this increase was not affected the chlorophyll synthesizing system and chlorophyllase
resulted from biouptake or biochemical metabolism by plants. activity. A decrease in the photosynthetic pigment content is well
When BCF values are investigated in the literature, it can be seen known from many other works concerning chromium stress in
that these values are higher in the studies performed at very low plants and it is attributed to the oxidative stress [21]. Augustynow-
metal ion concentrations than those in performed with moderate icz et al. [1] reported that very strong declining of photosynthetic
and high metal ion concentrations. Arora et al. [22] investigated activity was observed when plants exposed to chromium ions. They
the potential of Azolla spp. namely A. microphylla, A. pinnata and A. reported that 100 M Cr was the border concentration at which
liculoides to tolerate chromium ions in their growth environment some symptoms of the metal toxicity on the C. cophocarpa (water-
and bioconcentrate them. They reported that BCF values were in starwort) were found. They observed inhibition of growth and
the range 2434617 for three species. A. microphylla showed high- decrease in the photosynthetic pigment content at this concentra-
est bioconcentration potential, while its growth was affected to tion. 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 bioac-
cumulation 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 con-
centration 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 stud-
ies 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 bioup-
take capacity of L. minor on operational pH value and chromium
concentration of wastewater. It was proved that pilot system con-
taining 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. Conse-
quently, 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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