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Phytoremediation of Wastewater with

Limnocharis Flava, Thalia Geniculata and
Typha Latifolia in Constructed Wetlands
a a a
Alexander K. Anning , Percy E. Korsah & Patrick Addo-Fordjour
a
Department of Theoretical and Applied Biology, Kwame Nkrumah
University of Science and Technology, Kumasi, Ghana
Accepted author version posted online: 19 Sep 2012.Version of
record first published: 15 Nov 2012.

To cite this article: Alexander K. Anning , Percy E. Korsah & Patrick Addo-Fordjour (2013):
Phytoremediation of Wastewater with Limnocharis Flava, Thalia Geniculata and Typha Latifolia in
Constructed Wetlands, International Journal of Phytoremediation, 15:5, 452-464

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 International Journal of Phytoremediation, 15:452–464, 2013
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ISSN: 1522-6514 print / 1549-7879 online

DOI: 10.1080/15226514.2012.716098

PHYTOREMEDIATION OF WASTEWATER WITH

LIMNOCHARIS FLAVA, THALIA GENICULATA AND TYPHA
LATIFOLIA IN CONSTRUCTED WETLANDS

Alexander K. Anning, Percy E. Korsah, and Patrick

Addo-Fordjour
Downloaded by [University of Notre Dame Australia] at 06:38 22 April 2013

Department of Theoretical and Applied Biology, Kwame Nkrumah University of

Science and Technology, Kumasi, Ghana

Phytoremediation is thought to be the most sustainable wastewater treatment option for

developing countries. However, its application is often limited by unavailability of suitable
candidate species. In the present study, the potentials of Limnocharis flava, Thalia genic-
ulata and Typha latifolia for remediation of heavy metal contaminated wastewater with a
constructed wetland system were evaluated. The wetland consisted of three treatment lines
each planted with sufficient and equal number of a species. Duplicate plant and water sam-
ples were collected bi-monthly and analyzed for Fe, Cu, Zn, Pb, and Hg using the atomic
absorption spectrophotometer over a six month period. Bioaccumulation rates generally
increased over time and varied among plants for these metals, with Fe (456–1549 mg kg1
roots; 20–183 mg kg−1 shoot) being the most sequestered and Pb (1.2–7.6 mg kg−1 roots;
1.55–3.95 mg kg−1 shoot) the least. Translocation factors differed among the species but
generally remained stable over time. L. flava showed potential for hyperaccumulating Hg.
Removal efficiencies varied for the studied metals (∼ 20–77 %) and were generally related
to metal uptake by the plants. These results demonstrate the suitability of the species for
phytoremediation, and the usefulness of the technique as an option for improving irrigation
water quality in Ghana.
Supplemental materials are available for this article. Go to the publisher’s online edition of
International Journal of Phytoremediation to view the supplemental file.

KEY WORDS: bioaccumulation, heavy metal, removal efficiency, translocation factor

INTRODUCTION
Wastewater irrigation is a common practice among resource-poor vegetable farmers
in most developing countries (Agodzo et al. 2003; Chen et al. 2010). In many of these
countries, large volumes of untreated wastewater of both domestic and industrial origins
are often released into surface waters which are used for agricultural purposes (Keraita,
Drechsel, and Amoah 2003). Inocencio et al. (2003), for example, reported that wastewater
is used to irrigate about 900,000 hectares of farmland in developing countries. Studies
suggest that the farmers depend on polluted waters for irrigation due to limited access to
treated water, lack of legislation and rapid growth of the industry, among other factors
(Amoah et al. 2007; Arora et al. 2008).

Address correspondence to Alexander K. Anning, Department of Environmental and Plant Biology, 315
Porter Hall, Ohio University, Athens, OH 45791. E-mail: akanning.sci@knust.edu.gh

452
 PHYTOREMEDIATION OF WASTEWATER IN GHANA 453

In recent times, the poor quality of irrigation water has become a major source of
concern to many researchers and environmentalists due to the recognition of the associated
environmental and public health effects. Wastewater often contains significant amounts of
heavy metals including some that are toxic to both plants and animals (Gupta et al., 2008;
Massa et al., 2010). Consequently, elevated levels of toxic heavy metals are commonly
encountered in wastewater-irrigated crops and soils (Sharma, Agrawal, and Marshall 2007).
Heavy metals are also non-biodegradable, and tend to persist in the environment. Thus,
treatment of wastewater prior to its use in agriculture, remains an important option for
reducing the transfer of heavy metals through the food chain.
Conventional methods for treating wastewater, including biological (typically with
micro-organisms), chemical and physical, have been found to be difficult and inadequate
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for the large volumes of wastewater generated in most countries (Pescod 1992; Ghosh and
Singh 2005). As a result, phytoremediation, a group of plant-based technologies that use
naturally occurring or genetically engineered plants to reduce, remove, degrade, or immo-
bilize environmental toxins, is fast emerging as an alternative to conventional wastewater
treatment methods (Lasat 2002; Peer et al. 2005). This technology is relatively cheap and
considered the most suitable treatment option for developing countries, where access to
quality irrigation is limited (Ghosh and Singh 2005). Phytoremediation is commonly used
together with constructed wetlands—specifically constructed wetlands for pollution con-
trol and waste management, at places other than existing natural wetlands—due to their
sustainability, low maintenance, and energy costs (Maine et al., 2006). As noted by Krämer
(2010), many candidate plant species, including approximately 500 hyper-accumulators,
have been identified in different environments throughout the world. Nevertheless, re-
search to identify new species, particularly local species, continues because of its relevance
to phytoremediation success (Massa et al. 2010).
Vegetable growers in Ghana continue to rely on untreated wastewater for irrigation,
despite the documented occupational and health threats of the practice (Agodzo et al.,
2003; Keraita et al. 2003). Such wastewater typically contains large quantities of trace
metals (mainly Fe, Cu, Zn, Pb, and Hg), which tend to contaminate fresh-eating vegetables
(Cofie and Drechsel 2006; Sarpong 2007; Akoto, Bruce, and Darko 2008). In the absence
of sufficient conventional treatment plants (Agodzo et al. 2003; Amoah et al. 2003), phy-
toremediation can serve as a useful alternative for improving the quality of irrigation water.
Nevertheless, research to explore the potential of phytoremediation as a treatment option
in Ghana has been lacking. The present study was designed to assess the suitability of three
macrophytes, Limnocharis flava L. Buchenau, Thalia geniculata L. and Typha latifolia
L., for phytoremediation of metal contaminated wastewater using constructed wetlands. A
study in Ghana by Anning and Yeboah-Gyan (2007) showed that L. flava and T. genic-
ulata often share habitat with the common phytoremediation plant, T. latifolia (Phillips
et al. 2010), although the remediation potentials of the former two species have yet to be
assessed. The specific objectives of the study were to (1) determine the bioaccumulation
rates of Fe, Cu, Zn, Pb, and Hg accumulated by these plants, (2) compare their respective
translocation factors, and (3) determine their removal efficiencies for the studied metals in
the constructed wetlands.

MATERIALS AND METHODS

Location and Design of Constructed Wetland
A subsurface flow wetland system was constructed at the Kwame Nkrumah University
of Science and Technology (latitude 6◦ 35 N, longitude 1◦ 30 E), Kumasi for the study. The
 454 A.K. ANNING ET AL.

area is located within the moist semi-deciduous forest region of Ghana. Climate is humid
with mean annual rainfall and daily temperature of 1300 mm and 26 ◦ C respectively. The
soil is mainly sandy loam and is classified as Ferric Acrisol (Aikins and Afuakwa, 2010).
The KNUST campus is drained by the Wiwi and Sisa rivers. Water from these rivers,
polluted by influents from several laboratories and halls of residence in the University,
serves as irrigation water source for vegetable farmers in the surrounding communities.
The experimental wetland system (Figure S1) consisted of a storage tank, a sedimen-
tation tank, three parallel treatment lines and effluent tanks, all of which were connected
by 1 12 inch PVC pipes. Each treatment line had two serially-arranged rectangular cells (cell
dimension: 2.1 m length, 1 m width and 0.8 m depth). High-density polyethylene films
were placed at the bottom of each cell to prevent infiltration and soil contamination by
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percolation, while short-circuiting was prevented by inserting baffles underneath the cells.
The cells were filled with coarse sand (0.6–2 mm diameter) up to 0.2 m depth, followed
by 0.1 m layer of washed gravels (6.25 mm) as the filter medium and 30–40 mm diameter
stones at the inlet and outlet points to even out the flow rates of water. These substrate
layers were then covered with washed and sieved sand from Afisiyaso in Kumasi, which
had hydraulic conductivity of 368.87 cm3/d. All outlet points were covered with steel sieves
(4 mm pore size) to restrain large particles and debris from entering the connecting pipes.
Inflow discharge of 1 m3 per day was maintained to achieve a hydraulic residence time of
5–7 days. Total surface area and hydraulic loading rate required for proper functioning of
the wetlands were determined using plug flow reactor and first-order models (Kadlec and
Knight 1996). The wetlands were thoroughly checked for leakages, prior to transplanting.
A transparent nylon roof was used to prevent direct rainfall into the wetlands during the
early stage of the experiment (June-September) when this was a problem.

Transplanting of Test Plants and Stabilization of Wetland

Healthy-looking young plants of the three macrophytes—L. flava (yellow vel-
vetleaf, Limnocharitaceae), T. geniculata (arrowroot, Marantaceae) and T. latifolia (cattail,
Typhaceae)—were collected from nearby natural wetlands and immediately transplanted
into the constructed wetlands. The plants were grown separately with each treatment line
receiving one of the three species at a planting density of 5–6 transplants or cuttings
per m2. These gave off multiple plants (∼25–30 plants per cell) and allowed specimens
to be removed at the interval for the analysis. Wastewater was obtained from the Wiwi
River through the storage tank. Plants were allowed a two-month period to establish and
acclimate to their new environment in the wetland. Taxonomy and nomenclature of plants
followed Hutchinson and Dalziel (1963) and Akobundu and Agyakwa (1998).

Chemical Analyses
The concentrations of Fe, Cu, Zn, Pb, and Hg accumulated in the plants were analysed
after 12 weeks of growth in the wetlands. Starting from October 2010, two plants were
randomly collected, bi-monthly, from each of the six treatment cells into sterilized bags and
transported to the laboratory for analysis. These were thoroughly washed with tap water,
rinsed with distilled water and sorted into roots and shoots for a total of 48 samples per
species. After chopping into smaller pieces, the samples were weighed to determine their
fresh weights and then oven dried at 80 ◦ C for 72 h to determine their dry weights. Dried
samples were crushed in a mortar to pass through a 1 mm sieve and digested by dissolving
 PHYTOREMEDIATION OF WASTEWATER IN GHANA 455

2 g in a 250 ml glass beaker containing 10 ml nitric acid (65 %) and hydrochloric acid
(37 %) on a water bath for two hours, filtered, and then diluted to 50 ml with distilled
water (Jones and Case 1990). Analysis of heavy metals was performed in triplicate using
the atomic absorption spectrophotometer (AAS), according to standard protocols (APHA
1990). Shoot and root samples were analyzed separately. Concentrations of metals in the
plants were similarly determined prior to the start of the experiment, while concentrations
in the soil were determined using the aqua regia digestion protocol (USEPA 1996).
Duplicate water samples were collected monthly from each of the treatment cells and
the effluent tank for analysis. Samples were collected into sterilized sampling bottles and
transported immediately to the laboratory for the determination of dissolved heavy metals.
The samples were filtered using Whatman No. 41 filter paper, concentrated on a water bath
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and digested using 10 ml nitric acid and hydrochloric acid before heavy metal analysis with
the AAS.

Data Analyses
Bioaccumulation and translocation factors as were as removal efficiency were com-
puted according to the following formulae described by Shilev et al. (2008), Massa et al.
(2010), and Ganjo and Khwakaram (2010).

metal concentration in plant tissue

Bioaccumalation factor (BAF) = (1)
metal concentration in water
metal concentration in shoot
Translocation factor (TF) = (2)
metal concentration in root
inlet pollutant - outlet pollutant
Removal efficiency (RE) = × 100 % (3)
inlet pollutant
For the calculation of RE, concentration of metal in water collected at the point of
entry of each cell and the effluent tank were considered as the inlet pollutant and outlet
pollutants, respectively. Differences in bioaccumulation rates or BAFs, TFs, and REs were
statistically evaluated using the Kruskal-Wallis test, a nonparametric alternative to ANOVA.
The Shapiro-Wilk normality test was used to test the normality of the data before analysis.
Pairwise comparison tests, were performed using the kruskalmc command in the R package
“pgirmess.” The simple linear regression was used to assess the relationship between RE and
bioaccumulation rates/TFs, specifying RE as the dependent variable and bioaccumulation
rate/TF as the explanatory variable. The RE was log-transformed to stabilize its variance
before each analysis. Model diagnostics were performed using Q-Q plots and residual plots.
The R programming language (R Development Core Team 2011.) was used for all analyses.

RESULTS
Initial Concentrations of Heavy Metals in Water, Plant and Soil
Initial concentrations of all studied metals in the wastewater (except Cu) were within
safe limits for irrigation purposes (Table 1). Fe (with a mean concentration of 1.77 mg
L−1) was the most abundant element while Hg (0.05 mg L−1) was least abundant. Fe
(258.4 mg kg−1) was also the predominant metal in the soil while Pb (1.75 mg kg−1) was
the least abundant. Concentration of Fe in the three plants was about fivefold greater than
 456 A.K. ANNING ET AL.

Table 1 Initial concentrations of the studied metals in whole plant, soil and water samples together with their
safe limits and thresholds for toxicity and hyperaccumulation. In parenthesis are the standard errors. a Safe limits
for irrigation (Pescod 1992); Toxicity thresholds for bFe (Ganjo and Khwakaram 2010) and cother metals (Massa
et al., 2010); d hyperaccumulator criterion (Massa et al. 2010).

Concentration of heavy metal

Sample Fe Cu Zn Pb Hg

Plant
Limnocharis flava (mg kg−1) 468.0 (10.2) 19.38(2.9) 72.4(6.0) 4.2(0.8) 4.06(0.1)
Thalia geniculata (mg kg−1) 570.8 (11.6) 17.88(1.8) 86.0(7.9) 4.2(1.1) 5.16(0.0)
Typha latifolia (mg kg−1) 585.6 (9.3) 28.4 (2.7) 82.4(6.5) 5.6 (0.5) 5.14(0.5)
Soil (mg kg−1) 258.4 (0.1) 34.5 (0.00) 6.65 (0.05) 1.75 (0.05) 6.35(0.02)
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Water (mg l−1) 1.77 (0.52) 0.31 (0.01) 0.42 (0.00) 0.19 (0.01) 0.05 (0.00)
Safe limit (mg L−1)a 5.0 0.2 2.0 5.0 —
Toxic concentration (mg kg−1) 40–500b 20–100c 100–400c 30–300c 1–3c
Hyper-accumulation limit (mg kg−1)d 10000 1000 10000 1000 10

those of the other metals combined. T. latifolia had the largest amounts of Fe (585.6 mg
kg−1), Cu (28.4 mg kg−1) and Pb (5.6 mg kg−1), whereas T. geniculata recorded the
highest concentrations of Zn (86.0 mg kg−1) and Hg (5.16 mg kg−1). Concentration of
Hg in all species and that of Cu in T. latifolia, were within the toxicity limits (Table 1).
Initial concentrations of the metals in the species were, however, below their respective
hyperaccumulation limits.

Bioaccumulation Rates of Heavy Metals in the Constructed Wetlands

Bioaccumulation of heavy metals generally increased with time and varied among
the plant species (Figures 1 and S2). By the end of the study period, the plants had
accumulated more Fe than the rest of the studied metals combined. Rates of accumulation
were higher in the roots (456–1549 mg kg−1) than in the shoots (20–183 mg kg−1). Shoot
concentrations of T. geniculata (175 ± 16 mg kg−1) and L. flava (113 ± 4.53 mg kg−1)
differed significantly (P < 0.001) from that of T. latifolia (91 ± 16.27 mg kg−1). Pairwise
comparison indicated significantly higher accumulation rate in T. geniculata than in L.
flava. However, root concentrations of Fe did not differ statistically (P = 0.064) among
the species. Accumulation of Fe by the plants was below the hyper-accumulation threshold
(Table 1). The TF for Fe (< 0.4) was the lowest among the metals (Figure 2) and varied (P
< 0.001) among the species, with highest value found in T. geniculata.
T. latifolia accumulated more Cu (49.98 ± 15.11 mg kg−1 and 31.45 ± 1.23 mg
−1
kg respectively for shoots and roots) than the other plant species. Shoots concentration
of Cu only differed (P < 0.001) between T. latifolia and T. geniculata. On the contrary,
Cu accumulated in the roots varied among all but these species pair (P > 0.05). Like Fe,
bioaccumulation of Cu failed to reach the hyper-accumulation limit. Although the TF for
Cu differed (P = 0.001) among the species, it was greater than unity in all of them.
L. flava and T. latifolia accumulated significantly (P < 0.001) higher concentrations
of Zn than T. geniculata in both plant parts. Mean concentration of Zn in the shoot of T.
latifolia increased from 11.13 ± 2.18 to 32.9 ± 10.28 mg kg−1 between the start and end of
the experiment. Shoot concentration of the metal in T. geniculata was significantly lower
(P > 0.05) than in the other two species, and barely changed over time. Bioaccumulation
 PHYTOREMEDIATION OF WASTEWATER IN GHANA 457
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Figure 1 Mean concentrations of heavy metals accumulated by Limnocharis flava, Thalia geniculata and Typha
latifolia in the constructed wetland over the six month study period. P-values indicate statistical difference among
the plants (color figure available online).

rates in the roots were similar to those of the shoots, although slightly higher in L. flava than
in T. latifolia. Accumulation of Zn was also within the toxicity and hyper-accumulation
limits. TF of T. geniculata almost remained constant and above one for Zn, while those of
L. flava and T. latifolia fell below one over time, resulting in the difference (P = 0.017)
among the species.
Shoot and root concentrations of Hg in L. flava marginally exceeded the toxicity and
hyper-accumulation limits of 10 mg kg−1 (Figure 1) by the end of the study, and differed
statistically (P < 0.05) from those of the other plant species. Mean concentration of Hg
in T. latifolia (shoot) was lower than the other two species. Hg accumulated in the root of
T. geniculata (3.82–4.92 mg kg−1) was the lowest among the species. TFs differed (P <
0.001) among the species, and averaged about 1.5 for T. geniculata. TF of L. flava decreased
slightly over time but did not deviate much from one. On the contrary, TF of T. latifolia
was less than unity.
T. latifolia sequestered higher (P < 0.001) amounts of Pb than L. flava and T.
geniculata (Figure 1), but no significant difference was found between the latter two species.
Bioaccumulation rates of Pb (1.2–7.6 mg kg−1 in roots; 1.55–3.95 mg kg−1 shoots) were
 458 A.K. ANNING ET AL.
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Figure 2 Translocation factors of Limnocharis flava, Thalia geniculata and Typha latifolia over time. Error bars
represent standard errors. P < 0.05 value indicates statistical difference among the three species. A line above the
horizontal dashed line indicates the species effectively translocate heavy metals from its roots to the shoot (color
figure available online).

the lowest among the studied metals, and also fell below the toxic and hyperaccumulation
thresholds (Table 1). While the TFs of T. geniculata and T. latifolia fluctuated around one,
that of L. flava consistently stayed above one, indicating the effectiveness of the latter to
translocate Pb to its shoots. However, there were no statistical differences (P = 0.667)
among the TFs of the species.

Removal Efficiency of Heavy Metals

REs varied considerably among the plant species for the different metals studied
(Figure 3). However, no statistical differences (P > 0.05) were found between the REs of
the first and second cells or ponds. The plants were most efficient at removing Fe than the
other metals investigated, with RE ranging from 34.28 % in the wetland planted with T.
latifolia to 77.09 % in L. flava wetland. The L. flava and T. latifolia treatment lines differed
(P < 0.05) in their REs for Fe. For Hg, the L. flava wetland (51.61 %) again performed better
than those of T. latifolia (46.63 %) and T. geniculata (45.04 %), although the differences
were insignificant (P > 0.05). The T. latifolia wetland (51.88 %) was somewhat more
 PHYTOREMEDIATION OF WASTEWATER IN GHANA 459
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Figure 3 Removal efficiencies of Fe, Cu, Zn, Pb and Hg by wetlands planted with Limnocharis flava (LIMFLA),
Thalia geniculata (THAGEN) and Typha latifolia (TYPLAT) after treatment. Error bars represent standard errors
of the means. In each panel, different letters indicate statistical difference (P < 0.05).

effective at reducing the concentration of Zn than L. flava and T. geniculata wetlands,

which respectively, removed 44.45 % and 41.58 % of Zn. Again, these differences were
not significant (P > 0.05). The wetlands were generally less efficient in removing Pb and
Cu compared to the other metals. T. geniculata, T. latifolia and L. flava wetlands removed
30.26 %, 20.29 %, and 11.62 % of Pb respectively. The corresponding values for Cu were
9.39 %, 33.84 %, and 4.21 %.
Regression analysis showed strong relationship between RE and bioaccumulation
rate and/or TFs of the macrophytes (Table 2). Bioaccumulation of Fe, Zn, and Hg by L.
flava significantly influenced their removal from the wastewater. Conversely, REs of Cu and
Pb were not dependent on their accumulation by L. flava. The relationship between RE and
TF of L. flava was also significant for all metals but Pb. Accumulation of Fe, Cu, Zn, and Pb
by T. geniculata largely contributed to their removal, but translocation in this plant was only
related to the removal of Fe and Zn. The greatest number of significant associations between
RE and bioaccumulation/TF was observed for T. latifolia. Comparatively, bioaccumulation
in the roots of this plant showed greatest effect on RE.
 Downloaded by [University of Notre Dame Australia] at 06:38 22 April 2013

460
Table 2 Simple linear regression results showing the relation of removal efficiency with bioaccumulation rates and translocation factors of Limnocharis flava, Thalia geniculata
and Typha latifolia for the studied metals in a constructed wetland. Significant relations are indicated with bold face.

L. flava T. geniculata T. latifolia

Metal Plant part Coefficient R2 P-value Coefficient R2 P-value Coefficient R2 P-value

Bioaccumulation rate
Fe shoot 0.001 0.415 < 0.001 0.006 0.406 0.001 0.006 0.192 0.032
root 0.000 0.304 0.005 0.000 0.582 < 0.001 0.001 0.515 < 0.001
Cu shoot 0.034 0.107 0.119 0.008 0.001 0.871 0.005 0.094 0.155
root 0.003 0.000 0.943 0.132 0.228 0.018 0.039 0.738 < 0.001
Zn shoot 0.041 0.340 0.003 0.068 0.133 0.080 0.009 0.164 0.050
root 0.018 0.349 0.002 0.082 0.204 0.027 0.024 0.615 < 0.001
Pb shoot 0.121 0.011 0.664 1.240 0.516 < 0.001 1.152 0.415 0.001
root 0.126 0.013 0.639 0.242 0.051 0.290 0.253 0.362 0.002
Hg shoot −0.047 0.204 0.027 −0.146 0.088 0.158 −0.293 0.081 0.178
root −0.043 0.184 0.037 0.091 0.016 0.559 −0.286 0.248 0.013
Translocation factor
Fe −2.733 0.484 0.001 −2.349 0.656 < 0.001 2.125 < 0.001 0.996
Cu 1.306 0.373 0.001 −1.450 0.132 0.081 −0.000 0.021 0.504
Zn −0.729 0.445 0.004 −2.421 0.185 0.036 −0.227 0.089 0.018
Pb −0.735 0.010 0.669 0.558 0.052 0.282 −0.612 0.101 0.140
Hg −0.9683 0.5839 < 0.001 −0.558 0.082 0.175 −4.519 0.362 0.002
 PHYTOREMEDIATION OF WASTEWATER IN GHANA 461

DISCUSSION
Phytoremediation is a promising alternative to conventional wastewater treatment
methods in developing countries, as suggested by previous researchers (e.g., Ghosh and
Singh 2005). Successful application of the technique, however, depends on the identification
of plant species (particularly indigenous species) with the appropriate suite of characteristics
(Massa et al. 2010). In assessing the phytoremediation potential of a species, researchers of-
ten emphasize biomass production, bioaccumulation and tolerance of unusually high levels
of phytotoxins by the plant (Shah and Nongkynrih 2007; Krämer 2010). Hyperaccumulator
plants with the capacity to take up extremely high amounts of heavy metals (usually 0.1 %
to 3 % of dry weight) are even more desirable. In these respects, L. flava, T. geniculata, and
T. latifolia show considerable potentials as candidates for phytoremediation, as all three
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plants exhibited relatively high accumulation and translocation abilities in the constructed
wetland. Additionally, the species tolerated phytotoxic levels of Fe, Hg, and Pb, as no signs
of wilting were found during the study.
Differences in the bioaccumulation rates and TFs among the three species reflect the
abundance of the metals studied and the intrinsic abilities of the plants to sequester them.
Fe is one of the most abundant metals on earth, and several previous studies have provided
evidence of its predominance in water bodies in Ghana (e.g., Sarpong 2007; Akoto et al.
2008). As expected, Fe not only occurred in relatively higher concentrations in the water
and soil samples, but also accumulated in almost equal amounts in the three studied species.
Although, the plants accumulated more Fe than the rest of the metals, most of it remained
in the roots (Figures 1 and 2), agreeing with previous findings that T. latifolia (likewise L.
flava and T. geniculata) is good at extracting Fe from surrounding waters and generally
into the roots through immobilization in the rhizosphere (Ganjo and Khwakaram 2010).
The inability of the species to hyperaccumulate Cu, Pb and Zn is not surprising due to
the fact that only a few plants have these capabilities (Shilev et al. 2008). Nonetheless, T.
latifolia is a better accumulator of Cu and Pb than the other species, but less so with respect
to Zn. In a similar study, Ganjo and Khwakaram (2010) found the highest levels of Zn
and Cu in a congeneric species of T. latifolia (T. angustifolia). How these plants respond
to extreme concentrations of the heavy metals would be interesting to know, given the
observation by Kapourchal et al. (2009) that increasing Pb concentration in soils promoted
its accumulation in raddish (Raphanus sativus).
The prospect of Hg hyperaccumulation by L. flava is particularly interesting, con-
sidering the high toxicity and persistence of the metal in the environment, and the little
progress made so far in the search for hyperaccumulators of this metal (Su et al. 2009).
Besides exceeding the hyperaccumulation threshold, L. flava was effective at translocating
the metal from the roots to the shoots (Figure 2), which further demonstrates the capacity
of the plant to hyperaccumulate Hg (Krämer 2010). Su et al. (2009) reported comparatively
higher concentrations of Hg for the Chinese brake fern (Pteris vittata) and Indian mustard
(Brassica juncea) grown in hydroponic solutions, but also observed that Hg levels in the
plants were dependent on its concentration in the growth media. Thus, considering the
initially low concentration of Hg in the wastewater, the accumulation rate recorded in L.
flava is quite significant. These results, however, contrast the high root/shoot ratios of Hg
observed by Su et al. (2009) in their test plants.
The increasing pattern of bioaccumulation over the course of the study also demon-
strates the importance of time in the phytoremediation process. This finding supports
observation by Lai et al. (2010) that increasing the growth time promotes accumulation of
 462 A.K. ANNING ET AL.

heavy metals. Besides the time lag required for plant establishment, metal accumulation
involves several physiological and biological processes (Peer et al. 2005), which may take
considerable time to develop and become functional.
The removal of substantial quantities of heavy metals (particularly Fe, Zn and Hg)
from the wastewater by the wetlands provides further support for the use of phytoreme-
diation as a wastewater treatment method (Prasad and Freitas 2003; Peer et al. 2005).
Although removal of the metals was performed by the entire wetland system (cells and
the plants), significant relations between the REs and bioaccumulation rates or TFs of the
plants (Table 2) indicate that much of the reduction in concentration of the metals may be
attributed to their uptake by the plants. In this regard, the high RE of Fe (up to ∼ 34, 60,
and 77 % respectively by T. latifolia, T. geniculata, and L. flava) from the wastewater is
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plausible. Similar studies by Ganjo and Khwakaram (2010) and other researchers using
Typha species yielded between 33 and 91 % reduction in Fe, which agrees with the present
results. However, it is important to note that metal uptake is also a function of biomass pro-
duction, and this may confound the relationship between RE to bioaccumulation rate/TF.
For instance, the relatively higher rates of growth of T. geniculata and T. latifolia may have
compensated for their low bioaccumulation of Hg, resulting in the high RE of the metal.
Exactly why the plants could not efficiently reduce the concentrations of Pb and Cu in
the wastewater is not immediately clear. However, as Peer et al. (2005) observed, Pb has
extremely low solubility which poses a challenge for phytoremediation.
Heavy metal contamination of water bodies is considered less problematic in Ghana
because of scarcity of industries (Keraita et al. 2002; Cofie and Drechsel 2006). Nonetheless,
large quantities of wastewater, fraught with heavy metals, are often released into water
bodies in peri-urban areas of the country (McGregor, Thompson, and Simon 2002). For
example, high levels of Fe, Cu, Pb, Cd, As, and Hg above WHO recommended values for
irrigation purposes have recently been found in surface waters in the Kumasi metropolis
(Sarpong 2007; Akoto et al. 2008). In the present study, wastewater was collected from
only one source which happened to have low heavy metal loading. It is also worth noting
that heavy metals may build up to toxic thresholds in the environment and plants, even
when their concentrations in surface water are low (Gupta et al. 2008; Chen et al. 2010;
Sharma et al. 2007). Hence, there is a need for wastewater treatment prior to agricultural
use in order to reduce the associated risks in Ghana. Phytoremediation with constructed
wetlands clearly provides an efficient and cost-effective means for achieving this goal,
especially with the paucity of conventional treatment methods. The treatment system is
easy to construct and can be replicated in many places to give farmers access to clean water
for their irrigation needs. Identifying and evaluating more candidate species, particularly
indigenous plants, as was done in this study, would be an important starting point.

CONCLUSIONS
The present study demonstrates the potentials of L. flava, T. geniculata, and T.
latifolia for used in remediating heavy metal contaminated wastewater. L. flava and T.
geniculata compared fairly well with the widely used phytoremediation plant, T. latifolia,
in sequestering the studied metals. T. latifolia has high prospect for hyperaccumulating
Hg. The constructed wetland system was efficient at removing Fe, Zn, and Hg but less
so for Cu and Pb. REs were generally related to uptake of the metals by the plants. On a
broader scale, these results highlight the effectiveness of phytoremediation for improving
 PHYTOREMEDIATION OF WASTEWATER IN GHANA 463

the quality of irrigation water in Ghana. However, more studies integrating bioaccumulation
or TF assessment with biomass production of candidate species and metal accumulation in
irrigated crops are needed to facilitate the process.

ACKNOWLEDGMENTS
The authors thank the staff of the Department of Theoretical and Applied Biology,
KNUST, for research permits and logistical support. This research was supported by the
International Foundation for Science, Stockholm, Sweden, through a grant to Alexander K.
Anning.
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