Phytoremediation of municipal wastewater by using aquatic

plants.

INTRODUCTION
More than 70 % of the surface water is polluted by industrial and sewage effluents generated in
cities. Aanthropogenic activities releasing untreated sewage water is the major one which contains
high molecular weight compounds such as sugars, fats, oils, proteins obtained from domestic and
industrial wastes and can causes bad odour, colour, taste and algal growth [11]. Phytoremediation
is one of the best solutions for removing pollutants from the sewage and making it suitable for
reuse. Macrophytes-based wastewater treatment systems have several potential advantages
compared with conventional treatment systems and can act as biofilters in the wastewater
treatment technologies [12].
The conventional wastewater treatment process is inconvenient in the form of its operation and also
very costly due to its maintenance. Therefore, efforts are made for the use of natural process, which
can be used as an eco-friendly and effective source for treatment. Root zone is a bioremediation
process that uses various types of plants to remove, transfer, stabilize and destroy contaminants in
the wastewater. Plants can extract heavy metals, natural aromatic and hydrocarbon compounds
and man-made chemicals such as pesticides, herbicides, fungicides and antibiotics. This approach
is a promising clean up technology, which is both low-tech and low cost and reduces remedial cost,
restore habitat and clean up contamination [16].
The use of macrophytes such as Pistia and Eichhornia for phytoremediaion of wastewater polluted
by organic and inorganic pollutants was investigated by Elumalai et al., [8] and Fonkou et al., [9].
Similarly the capacity of vascular aquatic plants to assimilate nutrients from polluted waters is well
recognized viz, duckweed and Azolla offer potential alternatives for treating sewage. The use
of Azolla, an aquatic fern with high growth rate and productivity, seems to be very promising to
improve treated urban wastewater quality. The biosorption and bioadsorvent of N[O.sub.3.sup.-2],
P[O.sub.4.sup.-2] and S[O.sub.4.sup.-2] from sewage water using Azolla is well proven fact [14].
Azolla has been used extensively and effectively for green manure in rice field. Interest in the use of
this plant as a biological filter for the renovation of wastewater has increased nowadays [18].
Duckweed (Lemna minor) is a floating aquatic macrophyte, which occur worldwide on the surface
of nutrient rich fresh and brackish water. It is used in water quality improvement, to monitor heavy
metals, removal of nutrients, soluble salts, organic matter and suspended solids by accumulating
these compounds selectively. It has some unique physiological properties (small size, rapid growth
between pH 5 and 9 and vegetative propagation), which make it an ideal system

for phytoremediation [13]. The aquatic macrophytes work most efficiently when wastewater is
diluted in different ratios because the pollutants are also diluted, which become suitable for growth
and survival of these plants. The diluted wastewater is making favourable conditions
for phytoremediation.
Several researchers have worked on phytoremediation of municipalwastewater using various aquatic
plants. However the success ofimplementing this technology is depending on selection of
appropriateand most efficient plant species. The functioning of selected plantsystem depends on
type of contaminants in the wastewater and growth ofthe plants used. Present study was focused to
find out efficient andbetter functioning plant species for phytoremediation of sewage water
atdifferent levels of dilution and the testing was done to know the highlysuitable plant species for
wastewater treatment.
MATERIALS AND METHODS
Wastewater sample (about 120 litters) was collected from sewage treatment plants of Pune
Municipal Corporation located at Bopodi during the year 2012-13 and brought to the laboratory in
plastic containers for conducting the experiment. The aquatic plants selected for phytoremediation
viz Azolla pinnata and Lemna minor were collected freshly from natural pond at Horticulture
Research Station Ganesh Khind, Pune, and brought to the laboratory in plastic bags along with
water. These plants were cleaned properly to remove dirt and dust under tap water and stabilized
in laboratory conditions for 2-3 days to normalise their growth.
Factorial arrangement with randomized complete block design with three replications was used to
conduct the experiment at Department of Environmental Science, University of Pune, India.
Treatments included P0: no plant, P1: Azolla pinnata and P2: Lemna minor and ratios
of wastewater: distilled water were R0: 100 % wastewater, R1:75 % wastewater + 25 % distilled
water, R2: 50 % wastewater + 50 % distilled water and R3: 25 % wastewater + 75 % distilled water.
The wastewater after dilution with distilled water in the proportion as mentioned above was poured
into rounded, transparent plastic trough (18x20x20cm) having surface area 254 [cm.sup.2]. The
capacity of each plastic trough was about 5 litters. Selected plants, 5 g each of Azolla pinnata and
Lemna minor were inoculated separately in above containers as per the experimental design. The
laboratory conditions were maintained uniform throughout the experimental period (30 days).
All the physicochemical characteristics of wastewater samples (pre and post treatments) such as
pH, EC, TDS, N[O.sub.3.sup.-2], P[O.sub.4.sup.-2] and S[O.sub.4.sup.-2] were determined by
using standard methods (APHA, AWWA, WEF, 2005). The results were analyzed statistically by
using MSTATC computer software and a comparison of recorded data was done on the basis of
Duncan's multiple range tests at Alfa level 5%.
RESULTS AND DISCUSSION
pH:

The pH range during phytoremediation plays a key role and favourable pH for the growth of plants
varied according to species used for purification and dilution ratios. The results of
present investigation indicated that aquatic plants, dilution of wastewater and combination of both
(aquatic plants and diluted wastewater, 1:3) have caused significant reduction in pH value as
compare to initial stage (Fig. 1 and Table 1). Slightly alkaline pH was reported by Elumalai et al.,

[8]. The results of present study are in agreement with above work. According to Azeez and Sabbar,
[2] and Sengar et al., [15] release of CO2 during phytoremediation is positively correlated with
change in pH.
EC:
The electrical conductivity at final stage was reduced when compared to initial stage in all
treatments. Highest EC was recorded in absence of aquatic plants as compare to the presence of
Azolla and Lemna (Fig. 2 A). The EC was higher in municipal wastewater without dilution with DW,
while the lowest EC was observed with highest dilution ratio (1:3) (Fig. 2 B). In combination
treatment of dilution with DW and aquatic plants EC value was reduced as compare to no dilution
and no plants. The EC value goes on decreasing along with increasing dilution and use of plants.
The lowest EC was in treatment (1:3) dilution with Lemna (32 [micro]mohs/cm) followed by Azolla
(41 [micro]mohs/cm) (Table 1). The results clearly revealed reduction in EC in presence of plants as
well as with dilution of sewage wastewater. The results of present investigation are inconformity
with Chavan and Dhulap, [4]. They noted the reduction in EC by 32 to 80 % during sewage
treatment using Cana indica, which is promising emergent macrophytes for sustainable used
in wastewater treatment. The range of EC mostly depends on the concentration of various types of
soluble salts in wastewater. The decrease in EC during phytoremediation indicated the heavy
uptake of salts by both the plants used. Dipu et al., [6] and Dar et al., [5] recorded reduction in EC
with dilution and in presence of Pistia and Eichhornia species respectively.
TDS:
The total dissolved solids at final stage were decreased significantly in presence of Azolla and
Lemna as compare to their initial stage (Fig. 3 A). The TDS was decreased by increasing
dilution and the minimum value was in dilution treatment 1:3 (Fig. 3 B). The TDS value of municipal
wastewater varied with interaction between plant species and dilution ratios. The lowest value (21.3
mg/[l.sup.-]) (1: 3) was observed in presence of Lemna and highest dilution it was followed by
Azolla (26.7 mg/[l.sup.-]) (Table 1). Azeez and Sabbar, [2] reported very high reduction in TDS (48.9
%) in phytotreatment of wastewater with duckweed. They attributed the decrease in TDS to the
capacity of plants to take some organic and inorganic ions. Results of present study corroborates
with above findings. Dipu et al., [6] observed very high reduction in TDS for dairy effluents using
some aquatic macrophytes. Chavan and Dhulap, [4] also reported 25 to 50 % reduction in TDS
by using Cana indica. El-Kheir et al, [7] observed significant reduction in TDS, improving the
quality of domestic wastewater by using Lemna. Decrease in TDS reflects improvement in quality of
wastewater due to phytoremediation.
[FIGURE 1 OMITTED]
[FIGURE 2 OMITTED]
[FIGURE 3 OMITTED]
Nitrate:
Nitrates are commonly present in various forms in the wastewater and are important for plant
growth. The nitrate content in presence of Azolla and Lemna was reduced by 66.3 and 73.5 % as
compare control respectively (Fig. 4 A). The mean comparison of dilution effect showed lowest
N[O.sub.3.sup.-2] (15.11 mg/[l.sup.-]) in 1:3 dilution of wastewater as compare to no dilution (98.44
mg/[l.sup.-]) (Fig. 4 B). The impact of dilution and plant species clearly indicated

that N[O.sub.3.sup.-2] content was significantly lowered in presence of Azolla (3 mg/[l.sup.- ])

followed by Lemna (4 mg/[l.sup.-]) in (1: 3) ratio (Table 2). Azeez and Sabbar, [2] and El-Kheir et al.,
[7] reported the reduction of N[O.sub.3.sup.-2] in phytoremediation process by using Lemna.
According to Chavan and Dhulap, [3] the range of reduction in nitrates varied with type of plant
species used and they reported 42 to 70 % reduction in nitrate. The significant fall in nitrate
was attributed to its uptake for growth and metabolisms of the plant system used. Vipat et al, [17]
also reported the decreasing in N[O.sub.3.sup.-2] due to plant uptake and ions exchange.
Phosphate:
Phosphate is a major pollutant in wastewater and responsible for eutrophication. High
concentration of phosphate has negative effects on the structure of aquatic ecosystems [10].
Therefore it must be removed from wastewater during phytoremediation system before discharge
into environment. The results of present investigation showed that phosphate content was
decreased at final stage when compared with initial stage. The decreased was by 74 and 83 % with
Azolla and Lemna as compare to control respectively (Fig. 5 A). The influence of dilution of
municipal wastewater in different proportions indicated minimum phosphate content in the
treatment 1:3 dilution (2 mg/[l.sup.-]) as compare to the treatment no dilution (9 mg/[l.sup.-]) (Fig. 5
B). The impact of combination treatment (dilution of wastewater along with Azolla/Lemna) showed
that the minimum value was observed in the treatment 1:3 dilution and Lemna (0.3 mg/[l.sup.-])
followed by Azolla (0.7 mg/l) (Table 2). Many researchers such as Chavan and Dhulap, [3], Sengar et
al., [15] and Tripathi and Upadhyay, [16] recorded very high reduction in P[O.sub.4.sup.-2] content
in sewage water, dairy effluent and wastewater using different aquatic plants such as Panicum,
Algal species and Lemna respectively.
Sulphate:
Sulphate in very high concentration becomes the limiting factor for growth of plants and hence its
removal is essential during phytoremediation so that the treated wastewater can be reused
for agriculture purpose. The results of present study showed that S[O.sub.4.sup.-2] content was
decreased at final stage as compare to initial stage. Sulphate content at final stage was maximum
(37 mg/[l.sup.-]) in absence of plant species but it was highly reduced to minimum level (11
mg/[l.sup.-]) in presence of Lemna (Fig. 6 A). The mean comparison of ratios at final stage indicated
decrease in S[O.sub.4.sup.-2] with increasing dilution of sewage wastewater with DW and the
lowest S[O.sub.4.sup.-2] content was noted in 1:3 dilution ratios (Fig. 6 B). The interaction between
plant species used and the dilution ratios (1:3) revealed that minimum SO4-2 content
(1.3 mg/[l.sup.-]) was in presence of Lemna followed by Azolla (1.5 mg/[l.sup.-]) (Table 2). Chavan
and Dhulap, [3] observed more than 50 % reduction in the level of S[O.sub.4.sup.-2] using the
aquatic plant species Panicum. They noted that phytoremediation is most effective in highest
dilution and the treated water may be used for agriculture. They further stated that locally adapted
aquatic plants are more effective. Priya et al., [12] stated that wastewater treatment system using
aquatic macrophytes like Lemna, Azolla, Pistia etc has proved to be an efficient mechanism for
treating domestic as well as industrial wastewater.
[FIGURE 4 OMITTED]
[FIGURE 5 OMITTED]
[FIGURE 6 OMITTED]
Summary and conclusion:

Phytoremediation of municipal wastewater using aquatic macrophytes such as Azolla and Lemna
has proved to be a very convenient, highly effective and mostly nature friendly technology. It is low
cost and sustainable, protecting the environment from water pollution, which is very threatening to
the survival of human beings. From the results of present investigation it can be concluded that
Azolla and Lemna will serve the purpose of wastewater treatment in municipal areas which
are easily manageable. Amongst both them Lemna was more efficient for reducing almost all the
parameters studied in the state of dilution of wastewater (1: 3) as it is highly convenient for active
growth of these plants, which enable them to function at optimum level for improving the quality of
wastewater.
ARTICLE INFO
Article history:
Received 15 November 2013
Received in revised form 14
January 2014
Accepted 20 January 2014
Available online 25 February 2014
ACKNOWLEDGMENT
The authors express their sincere thanks to Head of Environmental Science Department, University
of Pune, Pune-7 for providing the research facility to conduct this experiment and for giving
inspiration, guidance as well as technical support.
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(1) Hossein Azarpira, (2) Pejman Behdarvand & (1) Kondiram Dhumal, (1) Gorakh Pondhe
(1) Department of Environmental Sciences, University of Pune, Pune-07-India.
(2) Department of Agricultural and Natural Resources, Ahvaz Branch, Islamic Azad University,

Ahvaz, Iran.
Corresponding Author: Hossein Azarpira, Department of Environmental Sciences, University of
Pune, Pune-07-India
E-mail: hazarpira912@gmail.com
Table 1: Interaction effect of plant species and ratios on pH, EC and TDS of wastewater during
phytoremediation Treatment pH EC ([micro]mohs/cm) Initial Final Initial Final P0R0 7.4 a 7.3 a
3555.0 a 3549.0 a P0R1 7.5 a 7.4 a 2682.0 b 2632.0 b P0R2 7.4 a 7.2 a 1793.0 c 1768.0 cd P0R3
7.3 a 7.2 a 881.3 d 866.7 e P1R0 7.5 a 7.1 a 2355.0 a 1938.0 c P1R1 7.5 a 7.0 a 2682.0 b 537.7
f P1R2 7.4 a 7.0 a 1793.0 c 176.0 g P1R3 7.2 a 7.1 a 881.3 d 41.0 g P2R0 7.3 a 7.2 a 3555.0 a
1592.0 d P2R1 7.4 a 7.3 a 2682.0 b 472.0 f P2R2 7.4 a 7.2 a 1793.0 c 121.3 g P2R3 7.3 a 7.2 a
881.3 d 32.0 g Treatment TDS (mg/[l.sup.-]) Initial Final P0R0 2355.0 a 2325.0 a P0R1 1805.0 b
1789.0 b P0R2 1153.0 c 1135.0 d P0R3 592.0 d 569.0 e P1R0 2355.0 a 1317.0 c P1R1 1805.0 b
358.3 f P1R2 1153.0 c 113.0 g P1R3 592.0 d 27.0 h P2R0 2355.0 a 1109.0 d P2R1 1805.0 b 325.0
f P2R2 1153.0 c 85.3 g P2R3 592.0 d 21.3 h Means with different letters are
significantly different at P = 0.05, using Duncan's Multiple Range Test. Table 2: Interaction effect
of plant species and ratios on N[O.sub.3.sup.-2], P[O.sub.4.sup.-2] and S[O.sub.4.sup.-2] content of
wastewater during phytoremediation Treatment N[O.sub.3.sup.-2] content (mg/[l.sup.-]) Initial
Final P0R0 153.3 a 143.3 a P0R1 118.0 b 116.3 b P0R2 76.0 c 70.7 d P0R3 43.0 d 38.3 e P1R0
153.3 a 80.0 c P1R1 118.0 b 35.0 e P1R2 76.0 c 6.7 g P1R3 43.0 d 3.0 g P2R0 153.3 a 71.0 d P2R1
118.0 b 19.0 f P2R2 76.0 c 4.0 g P2R3 43.0 d 4.0 g Treatment P[O.sub.4.sup.-2] content
(mg/[l.sup.-]) Initial Final P0R0 16.0 a 14.7 a P0R1 11.3 b 10.3 b P0R2 10.0 b 9.0 c P0R3 7.0 c
5.0 e P1R0 16.0 a 7.3 d P1R1 11.3 b 2.7 f P1R2 10.0 b 1.0 gh P1R3 7.0 c 0.7 hi P2R0 16.0 a 5.0
e P2R1 11.3 b 1.3 g P2R2 10.0 b 1.0 gh P2R3 7.0 c 0.3 i Treatment S[O.sub.4.sup.-2] content
(mg/[l.sup.-]) Initial Final P0R0 64.7 a 59.7 a P0R1 44.3 b 42.0 b P0R2 36.7 b 32.7 c P0R3 17.7 d
13.0 f P1R0 65.3 a 29.7 d P1R1 44.3 b 15.0 e P1R2 36.7 c 3.7 h P1R3 17.7 d 1.5 i P2R0 64.7 a
29.3 d P2R1 44.3 b 9.3 g P2R2 36.7 c 3.0 h P2R3 17.7 d 1.3 i Means with different letters are
significantly different at P = 0.05, using Duncan's Multiple Range Test.
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