International Journal of Phytoremediation

ISSN: 1522-6514 (Print) 1549-7879 (Online) Journal homepage: http://www.tandfonline.com/loi/bijp20

Aquatic macrophytes mediated remediation

of toxic metals from moderately contaminated
industrial effluent

Shweta Saraswat & Dr. J. P. N. Rai

To cite this article: Shweta Saraswat & Dr. J. P. N. Rai (2018) Aquatic macrophytes mediated
remediation of toxic metals from moderately contaminated industrial effluent, International Journal
of Phytoremediation, 20:9, 876-884, DOI: 10.1080/15226514.2018.1438359

To link to this article: https://doi.org/10.1080/15226514.2018.1438359

Published online: 06 Jun 2018.

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INTERNATIONAL JOURNAL OF PHYTOREMEDIATION
2018, VOL. 20, NO. 9, 876–884
https://doi.org/10.1080/15226514.2018.1438359

Aquatic macrophytes mediated remediation of toxic metals from moderately

contaminated industrial effluent
Shweta Saraswat, Ph.D.a and Dr. J. P. N. Rai, Ph.D.b
a
Environmental Sciences, College of Basic Science & Humanities, G. B. Pant University of Agriculture and Technology, U. S. Nagar, Uttarakhand, India;
b
Environmental Science, College of Basic Science & Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar, Udham Singh Nagar,
Uttarakhand, India

ABSTRACT KEYWORDS
The present study assessed Zn, Cr, Cd, and Pb removal efficiency of Colocasia esculenta, Hydrilla verticillata, aquatic macrophytes;
Phragmitis australis, Typha latifolia, and Spirodella polyrhiza from sewage-mixed industrial effluent. The bioconcentration factor;
fresh/dry weight and relative growth rate of each macrophyte decreased with increasing effluent heavy metals;
concentration. H. verticillata and C. esculenta exhibited better growth at 50% effluent over control. The phytoremediation;
translocation factor
maximum Zn, Cd, and Pb accumulation (1008.23, 28.03, and 483.55 mg/kg dry wt., respectively) was
recorded in C. esculenta, whereas Cr (114.48 mg/kg dry wt.) in H. verticillata at 100% effluent. Metal
accumulation in roots of all plants species was higher (
50%) initially with increasing effluent
concentration and later transferred to shoots. All plants exhibited BCF >1.0 for all heavy metals, highest
being for Zn (91.2) and Cd (75.2) in H. verticillata, for Cr (97.9) and Pb (103) in C. esculenta. Except S.
polyrhhiza, all other plants exhibited TF <1.0. Maximum removal efficiency of Zn was 82.8% by H.
verticillata, whilst that of Cr, Cd, and Pb by C. esculenta at 50% effluent, demonstrating wide applicability of
H. verticillata and C. esculenta for treatment of mixed industrial effluent having heavy metals.

Introduction and Salvinia herzogii to remove Cr (III) effectively than Cr (VI)

(Maine et al. 2016), thus emphasizing that locally adapted
Surface water contamination due to accelerated urban and
aquatic macrophytes have enormous ability to decontaminate
industrial discharge has led to various environmental and public
the polluted aquatic systems. However, the specificity of plants
health problems. The domestic wastewater along with untreated
to metal and its concentration must be addressed prior to their
or partially treated industrial effluent incorporate toxic recalci-
efficient application in phytoremediation technology.
trant trace metals in such water bodies which severely affects
In Uttarakhand, rapid industrialization and infrastructure
aquatic organisms and local inhabitants (Rai et al. 2002). Most
development at Integrated Industrial Estate, Pantnagar (IIE-
often, such wastewater is used for irrigation by local farmers
Pantnagar), has exposed the native water bodies to various
which not only affect soil fertility (Abou-Shanab et al. 2005;
industrial effluent inputs containing heavy metals and munici-
Oliveira and Pampulha 2006; Saraswat et al. 2007) but also bio-
pal wastewater from Rudrapur city. Such contamination must
accumulate the trace metals in plant products and ultimately
be removed by employing locally adapted metal accumulating
find its way into the food chain through biomagnification and
macrophytes to safeguard these aquatic systems. The present
thus warrant proper treatment. Some locally adapted aquatic
study was undertaken to assess the accumulation potential of
macrophytes growing in such contaminated water bodies are
Zn, Cr, Cd, and Pb in 5 locally adapted aquatic macrophytes
metal tolerant and highly efficient in removing toxic metals
(Colocasia esculenta, Hydrilla verticillata, Phragmitis australis,
from contaminated wastewater. Sahu et al. (2007) observed Spi-
Typha latifolia, and Spirodela polyrhiza) from sewage-mixed
rodela polyrhiza and Eichhornia crassipes as best accumulator of
industrial wastewater at microcosm level and to explore their
Cd and Zn in electroplating industrial wastewater. Rai (2008)
feasibility for in situ remediation of metals from industrial
reported Cd removal by 70–94% employing Azolla pinnata after
wastewater by determining bioconcentration factor, transloca-
13 days of treatment and Ha et al. (2009) recorded Zn and Cu
tion factor, and metal removal efficiency.
removal by 93% and 87%, respectively, using Eleocharis acicula-
ris after 10 days of exposure. Sun et al. (2013) also observed high
Zn, Cr, Cu, and Fe removal efficiency from electroplating waste- Materials and methods
water using Phragmitis australis, Acorus calamus, Typha
Study area
minima, and Lythrum salicaria. Further. Lemna minor was
reported to remove Cd and Pb by >80% from sewage-mixed The study area is located at Integrated Industrial Estate,
industrial wastewater (Bokhari et al. 2016) and Pistia stratiotes Pantnagar, Uttarakhand, India at a latitude between 28
590 5100

CONTACT Dr. Shweta Saraswat, Ph.D. shwetasara@gmail.com G. B. Pant University of Agriculture and Technology, Environmental Sciences, College of Basic
Science & Humanities, U. S. Nagar, 263145 India.
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/bijp.
© 2018 Taylor & Francis Group, LLC

and 29
010 1200 N and a longitude between 79
240 900 and
79
260 1500 E. The industrial area covers 1310 hectares of land
and harbours approximately 430 industries composed mainly
of electroplating, automobiles, battery manufacturing, food
production and processing, pharmaceuticals, plastic moulding,
PVC foam manufacturing, and FMCGs along with their allied
types (Banerjee and Srivastava 2010). Three sampling sites, i.e.,
S1, S2, and S3, located at a distance of 6.1 km, 2.5 km, and
4.5 km, respectively, from the centre of IIE-Pantnagar, were
selected for study. The S1 and S3 sites represent upstream and
downstream of Begul river, respectively, whereas S2 lies along
the border of the industrial estate carrying industrial wastewa-
ter discharge coupled with municipal sewage.
Collection and analysis of wastewater and aquatic
macrophytes
The acidified wastewater samples were stored in acid-washed
plastic container (5 L capacity) from each site and brought to
the laboratory along with associated plant samples and pre-
served at 2
C for further analysis. The physico-chemical
parameters (i.e., pH, electrical conductivity, COD, and heavy
metal content) of wastewater were determined in triplicate fol-
lowing APHA (1985). To analyse the metal content, i.e., Zn, Cr,
Cd, and Pb, 50 mL of wastewater was digested using 10 mL of
concentrated HNO3/HClO4 solution (3:1) at 80
C until it
became transparent followed by filtration and further dilution
up to 50 mL with double distilled water (APHA 1985). The
absorbance was recorded using Atomic Absorption Spectro-
photometer (AAS, GBC Avanta Ver. 1.33 (Australia)) fitted
with a specific lamp of a particular metal using appropriate drift
blanks. Analytical data quality of metals was ensured through
repeated analysis (n D 6) of standard reference samples and the
results were found to be within §2.05% of certified values. The
blanks were run with each set of samples in triplicate to check
the precision of the method.
The local aquatic macrophytes, viz., C. esculenta, P. australis,
S. polyrhiza, T. latifolia, H. verticilata, and C. indica were also
collected from all sites in bulk, washed thrice with distilled
water and oven dried at 80
C to constant weight. Zn, Cr, Cd,
and Pb content in plant samples were estimated after digestion
with HNO3/HClO4 (3:1, v/v) mixture at 80
C using AAS
(APHA 1985).
Microcosm study
To assess the Zn, Cr, Cd, and Pb removal efficiency of aquatic
macrophytes, the plants were collected from unpolluted water
bodies located on other side of Pantnagar. The plant samples
were rinsed with tap water immediately to remove any adhered
soil/sediment and were placed in plastic tanks filled with tap
water under natural sunlight for one week to let them adapt to
the new environment. The healthy acclimatized plants (50 g
fresh weight each) were kept in plastic troughs (5 L capacity
with equal volume and surface area) containing varied concen-
trations of wastewater (4 L). As such, the experimental set up
had 5 plant species with 3 wastewater concentrations, i.e., 25%,
50%, and 100% and 3 exposure durations, i.e., 10 days, 20 days,
and 30 days. Control experiment was carried out using tap
INTERNATIONAL JOURNAL OF PHYTOREMEDIATION 877
water with no wastewater addition. Altogether, 150 microcosms
(5 plant sp. £ 3 effluent concentrations £ 3 exposure duration
£ 3 replicates along with 15 microcosms as Control (5 plant
sp. £ 3 exposure duration)) were established. The water level
in the containers was maintained throughout the experiment
using deionized water. Experiments were performed in well
aerated glass house having 25/32
C night/day temperature and
relative humidity 65 § 5% in triplicate. The treated plants were
harvested after each experimental period and dried at 80
C.
Both plants and associated water samples were digested in
HNO3:HCIO4 (3:1, v/v) and metals were estimated using AAS
(APHA 1985). Values of metal accumulation have been given
after deducting metal concentrations of corresponding control
plants.
Relative growth of wastewater treated aquatic macrophytes
was calculated as the ratio of final fresh weight and initial fresh
weight after each treatment. Each plant’s dry weights (total dry
weight of all plant tissues), total growth (final dry weight¡ini-
tial dry weight), growth rate (g dry weight increase/total days of
exposure), relative growth rate (g dry weight increase/g initial
dry weight per day) were also determined to assess the toxicity
of contaminants in plant species. The dry weights were esti-
mated by extrapolation from original fresh weights and the
ratio of dry weight:fresh weight. The metal accumulation
potential of each macrophyte with respect to the metal concen-
tration in wastewater was also measured as bioconcentration
factor (BCF, Yoon et al. 2006).
mg of metal 6 g dry weight of root tissue
BCF D
mg of metal 6 g water
The translocation factor (TF) indicating metals transloca-
tion from root to shoot tissue (Yoon et al. 2006) was also
calculated as:
TF D ðMetal content in shootÞ=ðMetal content in rootsÞ
The tested plants’ efficiency to remove metals from wastewa-
ter was also calculated as:
Ci ¡ Ce
Metal removal efficiency .%/ D £ 100
Ci
where Ci and Ce are the initial metal concentration and final
metal concentration in the wastewater after 30 days of phytore-
mediation treatment, respectively.
Statistical analysis
Experimental data were presented as mean values § standard
error (S.E.). To verify the statistical significance of difference
among various treatments, the data were analyzed using t-test
and one-way analysis of variance as available in the SPSS statis-
tical package (Statgraphics Plus v. 11) and expressed at 0.05
and 0.01 probability levels.
878 S. SARASWAT AND J. P. N. RAI

Results Table 1. Physico-chemical characteristics of industrial effluent mixed domestic

wastewater collected from different streams nearby Sidcul, Pantnagar.
Wastewater characteristics and metal accumulation
*
Standards for
in plants Sites effluent discharge
Analysis of industrial wastewater collected from all sampling into inland
Parameters S1 S2 S3 surface water
sites revealed its slight acidic nature with high chemical oxygen
demand as compared to the prescribed Indian Standards pH 6.43§ 6.27 6.39 5.5-9.0
Conductivity (ms/cm) 149.8 176.2 184.6 —
(Table 1). The suspended solids, total solids, and inorganic Chemical oxygen demand 343 477 508 250
nutrients, i.e., NO3-N and PO4-P were also higher at down- (mg/L)
stream site. In general, each sampling site was observed con- Suspended solids (mg/L) 278.30 335.4 424.6 100
Total solids (mg/L) 420.78 579.7 663.7 500
taminated with heavy metals, viz., Zn, Cr, Cd, and Pb, being Zn NO3-N (mg/L) 8.25 16.48 19.21 10.0
and Pb conspicuously higher than permissible limits. Further, PO4-P (mg/L) 3.14 7.85 9.75 5.0
analysis of metals in naturally growing aquatic plants revealed Zn (mg/L) 7.28 14.46 16.7 5.0
Cr (mg/L) 0.75 1.66 2.4 2.0
that the metal accumulation was plant specific (Figure 1). In Cd (mg/L) 0.23 0.47 0.59 2.0
general, metal accumulation was higher in roots than shoots. Pb (mg/L) 2.41 5.39 6.58 0.1
Zn accumulation was significantly (P < 0.01) higher than other 
General standards for industrial effluent discharge into inland surface water as per
metals. Maximum Zn (1382.6 mg/kg dry wt.) and Cd (54.6 mg/ Schedule VI inserted by Rule 2(d) of the Environment (Protection) Second Amend-
kg dry wt.) were found in H. verticillata and P. australis, respec- ment Rules, 1993 notified vide G.S.R. 422(E) dated 19.05.1993, published in the
tively, whilst maximum Cr (239.76 mg/kg dry wt.) and Pb Gazette No. 174 dated 19.05.1993.
(122.4 mg/kg dry wt.) in T. latifolia. Among tested macro-
phytes, C. indica exhibited least metal accumulation in its tis-
sues, and thus not considered for further experiments. (18.35 mg/kg dry wt.). Further, the metal distribution pattern
in roots and shoots of studied plants depicted higher propor-
tion of metals in roots than shoots (Table 3). This trend was
Plant biomass and metal accumulation in microcosm
more conspicuous in Cd followed by Cr, Pb, and Zn, espe-
To interpret the plants’ behavior and tolerance towards heavy cially at 10 days wastewater exposure. However, with increas-
metals, their growth analysis was performed on both fresh and ing exposure duration metal content tend to increase in
dry weight basis. In general, the relative growth (based on fresh shoots particularly for Zn.
weight) of each plant species increased up to 50% wastewater
exposure and decreased at higher concentration (Table 2).
Metal bioconcentration and translocation factors
Among all, C. esculenta showed maximum relative growth
(1.84 g) followed by H. verticillata (1.69 g), T. latifolia (1.62 g), BCFs and TFs are the key measurements for determination of
P. australis (1.55 g), and S. polyrhiza (1.42 g). Further, the rela- metal accumulation potential of plants. Generally, plants exhib-
tive growth rate (RGR) of each plant species was also maximum iting TF and BCF >1 are considered good phytoremediators.
at 50% and minimum at 100% wastewater exposure (Table 2). Among various treatments, highest BCF was observed in case
The maximum RGR was recorded in C. esculenta (6.03 £ 10¡2 of 50% wastewater (Table 4). T. latifolia had highest BCF (91.2)
g/day), followed by H. verticillata (4.73 £ 10¡2 g/day) and T. for Zn followed by H. verticillata (90.7) and C. esculenta (90.3).
latifolia (4.30 £ 10¡2 g/day), whereas minimum (2.83 £ 10¡2 The highest BCFs for Cr (97.9) and Pb (103.0) were recorded
g/day) was in S. polyrhiza. in C. esculenta, whereas that for Cd was in H. verticillata (75.2).
Further, all plant species except S. polyrhiza exhibited metal
retention in roots than shoots as shown by TF <1 for different
Metal accumulation in plants
metals (Table 4). The TF values also varied among various
Results revealed that the metal accumulation was plant spe- treatments. The maximum TF for Zn and Pb (2.77 and 2.11,
cific. In general, metal accumulation was higher in roots than respectively) was calculated in S. polyrhiza and minimum in T.
shoots which increased with metal concentration in wastewa- latifolia (0.31 and 0.69, respectively) at 50% wastewater expo-
ter (Figure 2). The maximum Zn accumulation was observed sure. In case of Cr and Cd, the maximum TF (2.98 and 3.40,
in C. esculenta (1008.24 mg/kg dry wt.) followed in H. verticil- respectively) was also calculated in S. polyrhiza at 25% waste-
lata (943.92 mg/kg dry wt.) and T. latifolia (857.68 mg/kg dry water exposure.
wt.) whilst minimum in S. polyrhiza (727.74 mg/kg dry wt.).
The Pb accumulation recorded maximum in C. esculenta
Metal removal efficiency
(483.55 mg/kg dry wt.) followed by T. latifolia (347.76 mg/kg
dry wt.) and P. australis (317.84 mg/kg dry wt.) whilst mini- The metal removal efficiency of plants increased initially up to
mum in S. polyrhiza (262.42 mg/kg dry wt.). The maximum 50% wastewater concentration and reduced later (Table 5). An
Cr was found in H. verticillata (114.48 mg/kg dry wt.) fol- appreciably higher (>75%) metal removal was recorded from
lowed by T. latifolia (107.54 mg/kg dry wt.) and minimum in sewage-mixed industrial wastewater for all 5 plant species.
P. australis (85.2 mg/kg dry wt.). However, Cd accumulation Removal of Zn occurred maximally (82.8%) by H. verticillata
was found maximum in T. latifolia (28.49 mg/kg dry wt.) fol- followed by C. esculenta (79.9) at 50% wastewater exposure.
lowed by C. esculenta (28.03 mg/kg dry wt.) and H. verticillata The corresponding values for Cr and Pb (78.3% and 79.6%,
(27.03 mg/kg dry wt.) and minimum in P. australis respectively) was obtained by C. esculenta and Cd (72.9) by S.
 INTERNATIONAL JOURNAL OF PHYTOREMEDIATION 879

Figure 1. Metal accumulation (mg/kg dry weight) in root and shoot tissues of locally adapted aquatic macrophytes collected from streams nearby Sidcul, Pantnagar.
Mean values (§ SE) for a given plant species marked with asterisk () differed significantly at p < 0.05 () and p < 0.01 ().

polyrhiza, highlighting metal ion selectivity for removal by spe-
cific plant species.
Discussion
The acidic nature of sewage-mixed industrial wastewater might
be due to discharge of industrial effluents containing electro-
plating, metallurgical, and battery manufacturing wastes
(Table 1). The higher chemical oxygen demand at each site was
mainly due to addition of some domestic wastewater and sew-
age from upstream areas which finally joined the common
wastewater channel. Also, the higher Zn, Cr, Cd, and Pb
content in wastewater is primarily due to discharge of untreated
and/or partially treated metallurgical industries wastewater in
close vicinity of IIE-Pantnagar. Similar observations have been
recorded by Saraswat et al. (2007) for metallurgical industries
wastewater elsewhere.
Observations on metal accumulation in locally adapted
plant species revealed maximum accumulation of Zn
(Figure 1), which could be accounted for concentration-
dependent metal sequestration (Vyslouzilova et al. 2003).
However, maximum metal accumulation in roots of all
macrophytic species showed conformity with Chayapan
et al. (2015) and Madera-Parra et al. (2015), which may be
880 S. SARASWAT AND J. P. N. RAI

Table 2. Effect of varied concentrations of sewage-mixed industrial wastewater on different plants’ growth on exposure up to 30 days. Mean values (§S.E.) for a given
plant species marked with different letters (a and b) in the column differed significantly at P < 0.05 and P < 0.01, respectively, from control.
Plants Wastewater conc. (%) Initial dry wt. (g) Dry wt. after 30 days (g) Relative growth (g) Growth rate (£10¡2 g/day)

H. verticillata Control 2.05 § 0.05 3.33 § 0.12a 1.62a 4.27a

25 2.05 § 0.04 3.23 § 0.14a 1.58a 3.93a
50 2.05 § 0.02 3.47 § 0.09b 1.69b 4.73b
100 2.05 § 0.05 3.11 § 0.08c 1.52c 3.53c
P. australis Control 2.20 § 0.11 3.31 § 0.20a 1.50a 3.70a
25 2.20 § 0.06 3.39 § 0.11a 1.54a 3.97a
50 2.20 § 0.05 3.42 § 0.13b 1.55a 4.07b
100 2.20 § 0.02 2.86 § 0.07c 1.30b 2.20c
T. latifolia Control 2.08 § 0.08 3.24 § 0.09a 1.56a 3.87a
25 2.08 § 0.12 3.19 § 0.05a 1.53a 3.70a
50 2.08 § 0.03 3.37 § 0.13b 1.62a 4.30b
100 2.08 § 0.10 2.96 § 0.08c 1.42b 2.93c
S. polyrhiza Control 2.02 § 0.04 2.41 § 0.14a 1.19a 1.30a
25 2.02 § 0.09 2.47 § 0.10a 1.20a 2.37b
50 2.02 § 0.07 2.93 § 0.07b 1.42b 2.83c
100 2.02 § 0.03 2.86 § 0.05b 1.32c 2.13b
C. esculenta Control 2.15 § 0.05 3.39 § 0.11a 1.58a 4.13a
25 2.15 § 0.06 3.28 § 0.08a 1.71b 5.10b
50 2.15 § 0.10 3.56 § 0.15b 1.84b 6.03c
100 2.15 § 0.06 3.04 § 0.13c 1.55a 3.97a

due to the low mobility of metals from root to shoot cou-
pled with prevalent potential metal tolerance mechanisms
in roots (Kushwaha et al. 2016) leading to greater metal
retention in roots and slow translocation to shoots. Further,
plant behavior with respect to metal accumulation in micro-
cosm study also exhibited wastewater concentration and
exposure duration dependent metal accumulation, showing
conformity with the observation of Sahu et al. (2007) and
Kumar et al. (2013). Relatively sharp increase in metal
accumulation during initial 20 days of time threshold is due
to availability of abundant binding sites which get saturated
later and resulted in stabilized and/or reduced metal uptake
(Figure 2), as has also been observed by (Sousa et al. 2008;
Chen et al. 2016).

Table 3. Percent accumulation of Zn, Cr, Cd, and Pb in roots (R) and shoots (S) of different plant species exposed to varied wastewater concentrations for 10, 20, and
30 days.

Zn Cr Cd Pb
Plant Wastewater Plant
species* conc. (%) tissues 10 20 30 10 20 30 10 20 30 10 20 30

H. V 25 R 71.50 67.50 62.35 75.60 70.30 69.88 77.50 74.06 71.59 74.53 69.70 65.245
S 28.50 32.50 37.65 24.40 29.70 30.12 22.50 25.94 28.41 25.47 30.30 34.76
50 R 76.47 72.50 64.30 78.50 75.95 70.85 81.36 77.71 76.50 77.35 74.37 73.21
S 23.53 27.50 35.70 21.50 24.05 29.15 18.64 22.29 23.50 22.65 25.63 26.79
100 R 79.08 74.99 67.15 81.14 77.63 73.43 84.00 78.50 77.32 79.43 76.50 72.50
S 20.92 25.01 32.85 18.86 22.37 26.57 16.00 21.50 22.68 20.57 23.50 27.50
P. A 25 R 70.50 65.64 64.80 73.00 72.37 71.50 76.50 73.50 72.50 71.00 69.24 66.85
S 29.50 34.36 35.20 27.00 27.63 28.50 23.50 26.50 27.50 29.00 30.76 33.15
50 R 73.35 70.50 68.50 78.95 75.01 72.65 79.48 76.65 73.97 77.50 74.50 69.25
S 26.65 29.50 31.50 21.05 24.99 27.35 20.52 23.35 26.03 22.50 25.50 30.75
100 R 76.10 73.50 71.45 82.66 78.00 76.28 86.67 82.28 77.51 79.80 77.33 73.12
S 23.90 26.50 28.55 17.34 22.00 23.72 13.33 17.72 22.49 20.20 22.67 26.88
T. L 25 R 69.50 67.70 62.50 78.73 76.96 72.63 79.50 78.25 73.77 76.57 73.82 70.29
S 30.50 32.30 37.50 21.27 23.04 27.37 20.50 21.75 26.23 23.43 26.18 29.71
50 R 72.83 68.33 67.30 80.50 79.66 75.71 81.96 79.34 74.00 79.90 76.50 73.50
S 27.17 31.67 32.70 19.50 20.34 24.29 18.04 20.66 26.00 20.10 23.50 26.50
100 R 75.50 70.05 68.21 82.69 80.03 76.84 84.27 80.19 78.47 80.8 78.14 74.29
S 24.50 29.95 31.79 17.31 19.97 23.16 15.73 19.81 21.53 19.20 21.86 25.71
S. P 25 R 64.13 61.80 61.38 70.16 67.33 64.23 72.80 70.50 68.00 68.00 65.77 63.61
S 35.87 38.20 38.62 29.84 32.67 35.77 27.20 29.50 32.00 32.00 34.23 36.39
50 R 69.50 66.50 62.51 72.50 70.50 65.50 73.79 72.50 69.45 70.50 69.71 66.45
S 30.50 33.50 37.49 27.50 29.50 34.50 26.21 27.50 30.55 29.50 30.29 33.55
100 R 70.35 67.11 64.78 72.66 71.73 66.65 75.41 74.70 71.53 71.50 69.95 67.80
S 29.65 32.89 35.22 27.34 28.27 33.35 24.59 25.30 28.47 28.50 30.05 32.20
C. E 25 R 67.85 59.50 57.44 72.50 66.20 59.32 75.31 71.66 61.50 71.00 62.50 58.75
S 32.15 40.50 42.56 27.50 33.80 40.68 24.69 28.34 38.50 29.00 37.50 41.25
50 R 71.35 63.25 59.93 76.50 71.50 63.65 78.50 74.27 64.12 75.15 66.13 61.50
S 28.65 36.75 40.07 23.50 28.50 36.35 21.50 25.73 35.88 24.85 33.87 38.50
100 R 75.54 68.98 62.56 79.77 72.48 65.78 80.85 77.39 68.84 76.87 71.65 64.83
S 24.46 31.02 37.44 20.23 27.52 34.22 19.15 22.61 31.16 23.13 28.35 35.17

Plant species: H. V., Hydrilla verticillata; P. A., Phragmitis australis; T. L., Typha Latifolia; S. P., Spirodela polyrhiza; C. E., Colocasia esculenta.
 INTERNATIONAL JOURNAL OF PHYTOREMEDIATION 881

Figure 2. Zn, Cr, Cd, and Pb accumulation (mg/kg dry weight) in different plant species exposed to varied concentrations of sewage-mixed industrial wastewater up to
10, 20, and 30 days. Mean values (§SE) for a given plant species marked with asterisk () differed significantly at p < 0.05 () with their respective control and p < 0.01
() among various wastewater treatments. C represents Control.

The manifestation of toxicity with increasing wastewater
concentration led to reduction in plants growth and their rela-
tive growth rate, especially in 100% wastewater concentration
(Table 2), primarily due to metal-induced reduction in cell divi-
sion (Pastor et al. 2003; Yadav 2010) and negative interference
with important metabolic process such as photosynthesis (Pal
et al. 2006). Though, Zn is an essential element but its higher
concentration can cause reduction in root and shoot biomass
(Mendez et al. 2007), chlorophyll synthesis (Broadley et al.
2007), death of leaf tips and interference with P, Mg, and Mn
uptake (Rout and Das 2003). Besides, the non-essential metals
(i.e., Cd, Cr, and Pb) competing out with some essential ele-
ments (i.e., K, Ca, etc.) leading to reduced plant growth in con-
centrated wastewater, might have occurred in present study as
well. The results are in line with John et al. (2008) and Radic
et al. (2010) who reported harmful effects of Cd and Pb on
growth, chlorophyll content, carotenoid content, and enzy-
matic activities in Lemna polyrhiza and also with Hasan et al.
(2007), who reported reduction in fresh and dry weights, rela-
tive growth rate, and metal accumulation in E. crassipes on
882 S. SARASWAT AND J. P. N. RAI

Table 4. BCF and TF of heavy metals phytoremediated with different plants up to 30 days. Mean values given for each plant species marked with different letters in a col-
umn differed significantly at P < 0.05 and P < 0.01.
Bioconcentration factor (BCF)

Plant species Wastewater conc. (%) Zn Cr Cd Pb

H. verticillata 25 34.4 § 4.7a 40.1 § 5.6a 59.6 § 6.5a 60.8 § 5.4a

50 90.7 § 7.3b 58.1 § 8.3b 75.2 § 8.3b 58.2 § 4.2a
100 54.1 § 9.7c 49.8 § 5.9c 49.2 § 2.9c 40.0 § 3.8b
P. australis 25 63.5 § 5.2a 40.2 § 4.9a 44.5 § 3.8a 74.6 § 6.7a
50 82.0 § 10.2b 61.9 § 3.8b 74.9 § 6.9b 71.5 § 5.8a
100 50.8 § 6.3a 33.0 § 4.5a 28.9 § 2.2c 33.8 § 3.1b
T. latifolia 25 60.4 § 7.2a 58.9 § 7.9a 61.5 § 4.5a 69.9 § 7.2a
50 91.2 § 11.6b 63.8 § 8.1a 74.9 § 3.7a 81.7 § 9.3a
100 49.9 § 3.8c 42.4 § 3.4b 35.9 § 4.1b 56.8 § 6.7b
S. polyrhiza 25 25.2 § 2.7a 12.2 § 1.4a 21.2 § 1.8a 27.1 § 1.8a
50 27.0 § 3.1a 15.5 § 2.3a 24.0 § 1.8a 29.2 § 1.4a
100 20.9 § 2.5a 18.9 § 1.5a 22.2 § 1.3a 23.8 § 2.4a
C. esculenta 25 61.3 § 5.9a 56.7 § 3.5a 58.3 § 6.4a 76.7 § 8.5a
50 90.3 § 11.3b 97.9 § 5.2b 62.4 § 8.1a 103.0 § 9.6b
100 54.4 § 6.6a 33.0 § 2.4c 50.9 § 4.8a 67.4 § 8.2a
Translocation factor (TF)
H. verticillata 25 0.45 § 0.03a 0.41 § 0.06a 0.68 § 0.09a 0.38 § 0.05a
50 0.46 § 0.05b 0.36 § 0.04a 0.52 § 0.08b 0.69 § 0.05b
100 0.55 § 0.08b 0.42 § 0.06a 0.38 § 0.05b 0.48 § 0.03c
P. australis 25 0.41 § 0.04a 0.53 § 0.08a 0.62 § 0.07a 0.29 § 0.02a
50 0.29 § 0.03b 0.46 § 0.04b 0.18 § 0.01b 0.45 § 0.05b
100 0.43 § 0.05a 0.52 § 0.06a 0.41 § 0.03c 0.66 § 0.08c
T. latifolia 25 0.31 § 0.02a 0.21 § 0.03a 0.34 § 0.02a 0.42 § 0.04a
50 0.35 § 0.04a 0.42 § 0.05b 0.52 § 0.05b 0.45 § 0.03a
100 0.59 § 0.06b 0.48 § 0.06b 0.51 § 0.07b 0.48 § 0.06a
S. polyrhiza 25 2.33 § 0.09a 2.98 § 0.15a 3.40 § 0.23a 1.99 § 0.10a
50 2.77 § 0.08a 1.84 § 0.09b 3.33 § 0.19a 2.11 § 0.18a
100 2.05 § 0.1a 1.75 § 0.11b 2.17 § 0.25b 1.79 § 0.13a
C. esculenta 25 0.60 § 0.07a 0.51 § 0.06a 0.78 § 0.08a 0.47 § 0.05a
50 0.40 § 0.05b 0.28 § 0.03b 0.32 § 0.04b 0.26 § 0.03b
100 0.52 § 0.08a 0.37 § 0.04b 0.28 § 0.02b 0.49 § 0.08a

exposure of Zn and Cd either alone or in combination against
concentration gradient.
Apart from this, the appropriateness of a plant for metal
uptake from contaminated wastewater is often adjudged by its
BCF (Lin and Zhang 1990). BCF values over 1000 are generally
considered useful for metal accumulation (Jain et al. 1990).
Though, in present study, the BCF values of each plant species
were under 1000, yet these plants can be considered as a mod-
erate metal accumulator. Almost similar BCF values of Zn for
H. verticillata, T. latifolia, and C. esculenta (Table 3) indicated
equal plants’ potential for metals sequestration from wastewa-
ter and stabilize them into roots. Srivastava et al. (2009) and
Islam et al. (2017) also demonstrated BCF >1 of Zn in H. verti-
cillata and C. esculenta, respectively. Likewise, maximum BCF
values of Cr and Pb in C. esculenta reflected better metal accu-
mulation potential of these plants. In addition, T. Latifolia and
H. verticillata possessed better Cd accumulation potential in
roots than other plant species. The results were in accordance
with Sasmaz et al. (2008) who recorded BCF >1 and TF <1 for
Cd in T. latifolia growing in natural stream. Yabanli et al.
(2014) also demonstrated BCF >1000 in a submerged macro-
phyte Myriophyllum spicatum in order of Cr > Pb > Cd. How-
ever, among all plants, only S. polyrhiza exhibited TF >1.0
showed translocation of Zn, Cr, Cd, and Pb from root to shoot

Table 5. Zn, Cr, Cd, and Pb removal efficiency (%) of different plants exposed to varied metal concentrations of wastewater up to 30 days. The values given for each plant
species marked with different letters in a column differed significantly at P < 0.05 and P < 0.01.
Plants Wastewater conc. (%) Zn Cr Cd Pb

H. verticillata 25 74.1a 41.7a 59.3a 52.6a

50 82.8b 49.2a 72.9b 61.4a
100 52.3c 44.2a 42.4c 37.1b
P. australis 25 55.9a 38.3a 45.8a 60.5a
50 66.1b 56.7b 55.9b 65.0a
100 45.2c 31.3a 25.4c 35.1b
T. latifolia 25 49.5a 43.3a 52.5a 62.3a
50 77.0b 56.7b 52.5a 74.2b
100 49.5a 39.6a 33.9b 52.4c
S. polyrhiza 25 54.0a 30.3a 59.3a 50.8a
50 63.5b 37.5a 66.1a 57.4a
100 39.8c 32.5a 44.1b 44.1b
C. esculenta 25 61.4a 53.3a 56.1a 70.8a
50 79.9b 78.3b 62.5b 79.6a
100 51.7c 28.3c 40.7c 62.9b

(Table 4). Further, the aquatic plant’s potential for removing
metals from wastewater depends on 3 variables, viz., plant bio-
mass, BCF, and the volume of wastewater that requires remedi-
ation. On this ground, highest Zn removal efficiency (82.8%) in
H. verticillata, whilst Cr, Cd, and Pb removal efficiency (78.3%,
72.9%, and 79.6%, respectively) in C. esculenta (Table 5) clearly
demonstrated the wide applicability of these aquatic macro-
phytes as suitable metal miners for treating metal-laden waste-
water through natural and artificial constructed wetlands.
Conclusions
Experimental results depicted that H. verticillata and C. escu-
lenta possessed enormous potential for Zn, Cr, Cd, and Pb
removal from high voluminous industrial wastewater having
moderate concentration of metals by continuously passing
through a bed of these aquatic macrophytes. Owing to good
capacity of metal accumulation and high biomass potential,
these macrophytes can be employed as potential metal miners
for decontamination of metal-laden wastewater. Further, the
observation on plant subjectivity in metal removal as seen in
present study could be exploited for devising appropriate and
efficient phytoremediation technology for high voluminous
and moderate multi-metal contaminated wastewater.
Acknowledgments
The research facilities provided by G. B. Pant University of Agriculture &
Technology, Pantnagar, Uttarakhand are gratefully acknowledged.
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