International Journal of Scientific Research in Environmental Sciences (IJSRES), 1(4), pp. 63-70, 2013 Available online at http://www.ijsrpub.

com/ijsres ISSN: 2322-4983; 2013 IJSRPUB

Full Length Research Paper Phytoremediation of Heavy Metals from Urban Waste Leachate by Southern Cattail (Typha domingensis)
Amin Mojiri1*, Hamidi Abdul Aziz1, Mohammad Ali Zahed2, Shuokr Qarani Aziz3, M. Razip B. Selamat1
School of Civil Engineering, Universiti Sains Malaysia, 14300 Nibong Tebal, Penang, Malaysia 2 Department of Civil Engineering, Auburn University, Auburn, AL 36849, USA 3 Department of Civil Engineering, College of Engineering, Salahaddin University, Erbil, Iraq *Corresponding Author E-Mail: amin.mojiri@gmail.com Tel: +60169836292, Fax: +60345995999
Received 12 February 2013, Accepted 3 March 2013
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Abstract. The effectiveness of southern cattail (Typha domingensis) for phytoremediation of heavy metals from municipal waste leachate was investigated. Some plants were transplanted into pots containing 10 liters of mixed urban waste leachate and water (3/1 V: V) and aerated during experiments. Central composite design (CCD) and response surface methodology (RSM) were used in order to clarify the nature of the response surface in the experimental design and explain the optimal conditions of the independent variables. In the optimum conditions, the amount of removed Pb, Ni and Cd were 0.9725, 0.4681, and 0.3692 mg/kg, and Translocation Factor (TF) in 24, 48 and 72 h experiment were 1, 1.07, 1.00, 1.11, 1.32, 1.00, 1.5 1.20 and 1.02 for each heavy metal (Pb, Ni, and Cd) respectively. The findings show that Typha domingensis is an effective accumulator plant for phytoremediation of these heavy metals. Key words: Heavy metals; Phytoremediation; Typha domingensis; Waste leachate; Cattail

1. INTRODUCTION The use of plants for remediation of soils and waters polluted with heavy metals, has gained acceptance in the past two decades as a cost effective and noninvasive method (Mojiri, 2012). This approach is emerging as an innovative tool with great potential that is most useful when pollutants are within the root zone of the plants (top three to six feet). Furthermore, phytoremediation is energy efficient, cost-effective, aesthetically pleasing technique of remediation sites with low to moderate levels of contamination. The method of phytoremediation exploits the use of either naturally occurring metal hyper accumulator plants or genetically engineered plants (Setia et al. 2008). A variety of polluted waters can be phytoremediated, counting sewage and municipal wastewater, agricultural runoff/drainage water, industrial wastewater, coal pile runoff, landfill leachate, mine drainage, and groundwater plumes (Olgun and Galvn, 2010). A rising method for polluted area remediation is phytoextraction (Ok and Kim, 2007). Phytoextraction is the uptake of pollutants by plant roots and translocation within the plants. Pollutants are generally removed by harvesting the plants, and it has been recognized as an appropriated approach to remove pollutants from soil, sediment and sludge (Singh et al., 2011). Plants may play a vital role in metal removal through absorption, cation exchange, filtration, and chemical changes through the root.

There is evidence that wetland plants such as Typhalatifolia, Cyperus malaccensis and etc. can accumulate heavy metals in their tissues (Yadav and Chandra, 2011). Typha is often found close to water, in lakes, lagoons and riverine areas of numerous regions of the world, in America, Europe and Asia (Esteves et al., 2008). Typha is a highly flood-tolerant species with the capacity for internal pressurized gas flow to rhizomes through a well-developed aerenchyma system that provides oxygen for root growth in anaerobic substrates (Li et al. 2010). Southern cattail (Typha domingensis) is highly salt-tolerant and considered as the potential source of pulp and fiber (Khider et al., 2012). Dipu et al. (2012) conducted a study to determine the efficiency of an emergent wetland plant species Typha sp. and floating wetland macrophytes such as Pistia sp., Azolla sp., Lemna sp., Salvinia sp., and Eichhornia sp. in phytoremediation of various heavy metals with addition of a chelating agent such as EDTA. The aims of the study were to investigate the phytoremediation of heavy metals from urban waste leachate by Typha domingensis and optimization of process parameters using the response surface methodology (RSM). This technique has been employed for modelling and optimization of plant uptakes (Hu et al., 2006; Abhilash et al., 2011) as well as bioremediation (Zahed et al., 2010; Mohajeri et al.,

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2010) and phytoremediation (Cordova et al., 2011; Akinibile et al., 2012). 2. MATERIALS AND METHODS 2.1. Sample Preparation The plants were transplanted into pots containing 10 liters of mixed urban waste leachate and water (mixed 75 percentages of waste located with 25 percentage of

water; V: V), and aeration was done in 2011. Central composite design and response surface methodology were used in order to clarify the nature of the response surface in the experimental design and explain the optimal conditions of the independent variables. Different number of Typha domingensis transplanting in each pot (2 to 4 Typha domingensis) and different lengths of time for taking samples (24 to 72 hours) were used.

pH

EC (dS m-1) 0.23 28.72

N (mg/L)

7.00 5.84

Table 1: Waste leachate and water properties BOD5 Fe Mn Zn Pb (mg/L) (mg/L) (mg/L) (mg/L) (mg/L) Water ND ND ND ND ND Urban Waste leachate 0.71 27.18 80.013 16.011 17.11 2.218

Ni (mg/L) ND 1.092

Cd (mg/L) ND 0.925

ND: Not Detected, MDL: 10 g/L

Run

1 2 3 4 5 6 7 8 9 10 11 12 13

Table 2: Experimental variables and results for the removal metals Variables Response A: number of plants B: Time for taking Amount of Pb Amount of Ni transplanting samples removed. removed. (hours) (mg/kg) (mg/kg) 2 48 0.800 0.343 4 48 0.901 0.420 3 48 0.870 0.383 2 24 0.703 0.380 3 48 0.881 0.397 4 72 1.130 0.589 3 48 0.882 0.398 3 72 0.997 0.492 2 72 0.798 0.372 3 48 0.879 0.399 3 48 0.889 0.390 4 24 0.802 0.341 3 24 0.76 0.437

Amount of Cd removed. (mg/kg) 0.300 0.341 0.314 0.282 0.322 0.447 0.322 0.386 0.300 0.327 0.318 0.301 0.310

2.2. Laboratory Analysis The plant tissues were prepared for laboratory analysis by Wet Digestion method (Campbell and Plank, 1998). Extractable lead (Pb), nickel (Ni) and cadmium (Cd) in waste leachate and plant tissues were carried out using a flame atomic absorption spectrometer (Varian Spectra 20 Plus, Mulgrave, Australia) in accordance to the Standard Methods (APHA, 2005). Waste leachate and water properties are shown in Table 1.
Y

2.3. Statistical Analysis Central composite design (CCD) and Response surface methodology (RSM) were employed in order to clarify the nature of the response surface in the experimental design and elucidate the optimal conditions of the independent variables. CCD was established through Design Expert Software (6.0.7). The behavior of the system is described through equation 1 an empirical second-order polynomial model:

i
i 1

i 1

x
ii

i 1

i j 1

xx
ij i

ij

(1)

where Y is the response; Xi and Xj are the variables; 0 is a constant coefficient; j, jj, and ij are the interaction coefficients of linear, quadratic and second-order terms, respectively; k is the number of study factors; and e is the error.

The results were completely analyzed by analysis of variance (ANOVA) in the Design Expert Software. Number of Typha domingensis transplanting (2, 3, and 4) and times for taking samples (24, 48, and 72 hours) were used. To carry out an adequate analysis, three

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International Journal of Scientific Research in Environmental Sciences (IJSRES), 1(4), pp. 63-70, 2013

dependent parameters (reducing lead, nickel, and cadmium concentration in leachate) were measured as responses (Table 2). Descriptive statistical analysis including mean comparison of Pb, Ni and Cd accumulation in the roots and shoots of the plants using Duncans Multiple Range Test (DMRT) was conducted using the SPSS software.

3. RESULTS AND DISCUSSIONS Waste leachate properties before the experiment, the results of the experiments, ANOVA results for response parameter, and comparing the means of Pb, Ni and Cd accumulation in Typha domingensis roots, and shoots are shown in Tables 2.

Response Pb Removal Ni Removal Cd Removal

Table 3: ANOVA results for response parameter Final equation in terms of actual Prob. R2 Adj.R2 SD factors 0.7321-0.0278A-0.003B-0.003AB 0.0001 0.9520 0.9360 0.027 0.05318+0.0788A-0.0152B0.0273A2-0.0050B2-.003AB 0.3432+0.0157A-0.003B-0.005B0.003AB 0.0001 0.0001 0.9554 0.9623 0.9235 0.9354 0.018 0.011

CV 3.15 4.46 3.27

PRESS 0.025 0.020 0.030

Prob.LOF 0.0033 0.0113 0.0222

Prob.: Probability of error; R2: Coefficient of determination; Ad. R2: Adjusted R2; Adec. P.: Adequate precision; SD: Standard deviation; CV: Coefficient of variance; PRESS: Predicted residual error sum of square; Prob. LOF: Probability of lack of fit Where A is number of Typha domingensis transplanting, and B is time for taking samples

Metals(mg/L) Pb Ni Cd

Table 4: Comparison the heavy metals TF in Typha domingensis after 24, 48 and 72 hours Time(h) Plants TF Time(h) Plants TF Time(h) Plants Root Shoot Root Shoot Root Shoot 24 0.291a+ 0.292a 1.00 48 0.401a 0.448f 1.11 72 0.700a 0.739f 0.102b 0.110b 1.07 0.209b 0.277g 1.32 0.312b 0.375g 0.096c 0.096c 1.00 0.128c 0.129h 1.00 0.293c 0.300h
+ Numbers followed by same letters in each column are not significantly (P<0.05) different according to the DMR test

TF 1.05 1.20 1.02

In this work, the RSM was used for analyzing the correlation between the variables (number of Typha domingensis transplanting and the lengths of time for taking samples) and the important process response (the amount of removed Pb, Ni, and Cd). Predicted vs. actual values plot for metal removals are shown in Figures 1. Considerable model terms were preferred to achieve the best fit in a particular model. CCD permitted the development of mathematical equations where predicted results (Y) were evaluated as a function of the number of Typha domingensis transplanting (A) and the lengths of time for taking samples (B). The results were computed as the sum of a constant, two first order effects (terms in A and B), one interaction effect (AB), and two second-order effects (A2 and B2), as shown in the equation (Table 3). The results were analyzed by ANOVA to determine the accuracy of fit. The model was significant at the 5% confidence level because probability values were less than 0.05. The lack of fit (LOF) F-test explains variation of the data around the modified model. LOF would be significant, if the model did not fit the data well. Generally, large probability values for LOF (>0.05)

explained that the F-statistic was insignificant, implying a significant model relationship between variables and process responses. 3.1. Lead (Pb) Removed The amount of removed Pb ranged from 0.703 mg/kg (2 plants transplanting, and 24 hours of time for taking samples) to 1.130 mg/kg (4 plants transplanting, and 72 hours of time for taking samples). The phytoremediation of Pb increased when the number of plants transplanting and time for taking samples were increased. 3.2. Nickel (Ni) Removed The amount of removed Ni ranged from 0.341 mg/kg (3 plants transplanting, and 24 hours of time for taking samples) to 0.589 mg/kg (4 plants transplanting, and 72 hours of time for taking samples). The phytoremediation of Ni increased when the number of plants transplanting and time for taking samples were increased.

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(a)

(b)

(c) Fig. 1: The design expert statistical plots - predicted versus actual plot: (a) Pb, (b)Ni, (c) Cd

3.3. Cadmium (Cd) Removed The amount of removed Cd ranged from 0.282 mg/kg (2 plants transplanting, and 24 hours of time for taking samples) to 0.447 mg/kg (4 plants transplanting and 72 hours of time for taking samples). The phytoremediation of Cd increased when the number of plants transplanting and time for taking samples were increased. 3.4. Uptake of Heavy Metals by Plant Soluble metals can enter into the root symplast by crossing the plasma membrane of the root endodermal cells, or they can enter the root apoplast through the

space between cells. While it is possible for solutes to travel up through the plant by apoplastic flow, the more efficient method of moving up the plant is through the vasculature of the plant, called the xylem. To enter the xylem, solutes must cross the Casparian strip, a waxy coating, which is impermeable to solutes, unless they pass through the cells of the endodermis. Therefore, to enter the xylem, metals must cross a membrane, probably through the action of a membrane pump or channel. Once loaded into the xylem, the flow of the xylem sap will transport the metal to the leaves, where it must be loaded into the cells of the leaf, again crossing a membrane. The cell types where the metals are deposited vary between hyper-accumulator species (Peer et al., 2005).

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International Journal of Scientific Research in Environmental Sciences (IJSRES), 1(4), pp. 63-70, 2013

(a)

(b)

(c) Fig. 2: The 3D surface plots of heavy metal removal: (a) pb, (b) Ni, (c) Cd

Metal accumulating plant species can concentrate heavy metals like Cd, Zn, Co, Mn, Ni, and Pb up to 100 or 1000 times more than those taken up by nonaccumulator (excluder) plants. The uptake performance by plant can be greatly improved (Tangahu et al., 2011). The concentrations of lead (ppm) in the roots of Typha domingensis were 0.291, 0.401, and 0.700, and in the shoots of Typha domingensis were 0.292, 0.448, and 0.739, after 24, 48, and 72 hours, respectively. The lead is not necessary for plant growth and considered as contaminated at the concentration of 30300 g g-1 in plant tissues Mojiri (2012). The concentrations of nickel (ppm) in the roots of Typha domingensis were 0.102, 0.209, and 0.312, and

in the shoots of Typha domingensis were 0.110, 0.277, and 0.375, after 24, 48, and 72 hours, respectively. The concentrations of cadmium (ppm) in the roots of Typha domingensis were 0.096, 0.128, and 0.293 and in shoots of Typha domingensis were 0.096, 0.129, and 0.300, after 24, 48, and 72 hours, respectively. 3.5. Translocation factor (TF) The efficiency of phytoremediation can be quantified by calculating translocation factor. The TF expresses the capacity of a plant to store the MTE in its upper part. This is defined as the ratio of metal concentration in the upper part to that in the roots (Chakroun et al., 2010). The translocation factor indicates the efficiency

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of the plant in translocating the accumulated metal from its roots to shoots. It is calculated as follows (Padmavathiamma and Li, 2007).

Translocation Factor (TF )

CShoot C Root

(2)

where Cshoot is the concentration of the metal in plant shoots and Croot is the concentration of the metal in plant roots. Based on Table 4, translocation factors (TF) were more than 1 in all treatments. A translocation factor value greater than 1 indicates the translocation of the metal from root to above-ground part (Jamil et al. 2009). According to Yoon et al. (2006), only plant species with TF greater than 1 have the potential to be used for phytoextraction. 4. CONCLUSION Phytoremediation of heavy metals from urban waste leachate by Typha domingensis was studied. CCD and RSM were used in the design of experiments, statistical analysis and optimization of the parameters. The factors were number of Typha domingensis transplanting (2, 3, and 4) and time for taking samples (24, 48, and 72 hours); while the responses were removals of Pb, Ni and Cd. The findings clarified that the Typha domingensis is an effective accumulator plant for phytoremediation of Pb, Ni and Cd. Statistical analysis via Design Expert Software (6.0.7) showed that the optimum conditions for the number of Typha domingensis transplanting and the time for taking samples were 3.33 and 61.90 hours, respectively. For the optimized factors, the amount of removed pollutants Pb, Ni and Cd (ppm) were 0.9725, 0.4681, and 0.3692 mg/kg, respectively. Acknowledgements The authors would like to acknowledge the University Sains Malaysia (USM) for their supports. REFERENCES Abhilash PC, Srivastava S, Srivastava P, Singh B, Jafri A, Singh N (2011). Influence of rhizospheric microbial inoculation and tolerant plant species on the rhizoremediation of lindane. Environmental and Experimental Botany, 74: 127-130. Akinbile CO, Yusoff MS, Shian LM (2012). Leachate Characterization and Phytoremediation Using Water Hyacinth (Eichorrnia crassipes) in Pulau Burung, Malaysia. Bioremediation Journal, 16(1): 9-18.

APHA (2005) Standard Methods for Examination of Water and Wastewater. 21th ed. American Public Health Association, Washington, DC, USA. Campbell CR, Plank CO (1998). Preparation of plant tissue for laboratory analysis. P 37-49. In Y.P. Kalra (ed) Handbook of Reference Method for Plant Analysis. CRC Press, Boca Raton, FL. Chakroun HK, Souissi F, Bouchardon JL, Souissi R, Moutte J, Faure O, Remon E, Abdeljaoued S (2010). Transfer and accumulation of lead, zinc, cadmium and copper in plants growing in abandoned mining-district area. African Journal of Environmental Science & Technology, 4(10): 651-659 Crdova FC, Len AG, Reyes RG, Gonzlez MG, Soto E (2011). Response surface methodology for lead biosorption on Aspergillus terreus. Int J Environ Sci Tech, 8(4): 695-704. Dipu S, Kumar AA, Thanga SG (2012). Effect of Chelating Agents in Phytoremediation of Heavy Metals. Remediation Journal, 22(2): 133-146. Esteves BS, Enrich-Prast A, Suzuki MS (2008). Allometric relations for Typha domingensis natural populations. Acta Limnol Bras, 20(4): 305-311. Hu ZH, Yu HQ, Zheng JC (2006). Application of response surface methodology for optimization of acidogenesis of cattail by rumen cultures. Bioresource technology, 97(16): 2103-2109. Jamil S, Abhilash PC, Singh N, Sharma PN (2009). Jatropha curcas: A potential crop for phytoremediation of coal fly ash. J Hazard Mater, 172: 269-275. Khider TO, Omer S, Taha O (2012). Alkaline Pulping of Typha domingensis stems from Sudan. World Applied Sciences Journal, 16 (3): 331336. Li S, Lissner J, Mendelssohn LA, Brix H, Lorenzen B, McKee KL, Miao S (2010). Nutrient and growth responses of cattail (Typha domingensis) to redox intensity and phosphate availability. Annals of Botan 105: 175184. Mohajeri L, Aziz HA, Zahed MA, Mohajeri S, Kutty SRM, Isa MH (2011). Response surface analysis and modeling of n-alkanes removal through bioremediation of weathered crude oil. Water Sci Technol 9(63.4): 9. Mojiri A (2012). Phytoremediation of heavy metals from municipal wastewater by Typha domingensis. African Journal of Microbiology Research, 6(3): 643-647. Ok YS, Kim JG (2007). Enhancement of Cadmium Phytoextraction from Contaminated Soils with Artemisia princeps var. orientalis. Journal of Applied Sciences, 7(2): 263-268.

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Olgun EJ, Galvn GS (2010). Aquatic phytoremediation: Novel insights in tropical and subtropical regions. Pure Appl Chem, 82(1): 27-38. Padmavathiamma PK, Li LY (2007). Phytoremediation technology: hyper accumulation metals in plants. Water Air Soil Pollut, 184: 105-126. Peer WA, Baxter IR, Richards, EL, Freeman, JL, Murphy AS (2005). Phytoremediation and hyperaccumulator plants. The University of Chicago, The Science Behind Genetically Modified Organisms, pp: 43. Setia RC, Kuar N, Setia N, Nayyar H (2008). Heavy Metal Toxicity in Plants and Phytoremediation. Crop Improvement: Strategies and Applications, I.K. International Publishing House Pvt. Ltd., New Delhi, pp 206-218. Singh D, Gupta R, Tiwari A (2011). Phytoremediation of Lead from Wastewater Using Aquatic Plants. International Journal of Biomedical Research, 2(7): 411421.

Tangahu BV, Abdullah SRS, Basri H, Idris M, Anuar N, Mukhlisin M (2011). A Review on HeavyMetals (As, Pb, and Hg) Uptake by Plants through Phytoremediation. International Journal of Chemical Engineering, Article ID 939161, 31 pages. Yadav S, Chandra R (2011). Heavy metals accumulation and ecophysiological effect on Typha angustifolia L. and Cyperus esculentus L. growing in distillery and tannery effluent polluted natural wetland site, Unnao, India. Environ. Earth Sci, 62: 12351243. Yoon J, Cao X, Zhou Q, Ma LQ (2006). Accumulation of Pb, Cu, and Zn in native plants growing on a contaminated Florida site. Sci Total Environ, 368: 456-464. Zahed MA, Aziz HA, Mohajeri L, Mohajeri S, Kutty SRM, Isa MH (2010). Application of statistical experimental methodology to optimize bioremediation of n-alkanes in aquatic environment. J Hazard Mater, 184(1): 350-356.

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Amin Mojiri is a PhD candidate in environmental engineering, School of Civil Engineering, Universiti Sains Malaysia (USM), Pulau Pinang. He is fellowship holder and research assistant at the School of Civil Engineering (USM). He is a member of Young Researchers Club, Islamic Azad University, Iran. He is editor and reviewer of some international journals. His area of specialization is waste management, waste recycling, wastewater treatment, wastewater recycling, and soil pollutions.

Dr Aziz is a Professor in environmental engineering at the School of Civil Engineering, Universiti Sains Malaysia. Dr. Aziz received his Ph.D in civil engineering (environmental engineering) from University of Strathclyde, Scotland in 1992. He has published over 200 refereed articles in professional journals/proceedings and currently sits as the Editorial Board Member for 8 International journals. Dr Aziz's research has focused on alleviating problems associated with water pollution issues from industrial wastewater discharge and solid waste management via landfilling, especially on leachate pollution. He also interests in biodegradation and bioremediation of oil spills.

Dr. Mohammad Ali Zahed received PhD in Environmnetal Engineering from Universiti Sains MAlasia (USM). He is editor and reviewer some international journals.

Dr. Shuokr Qarani Aziz is a lecturer in the Civil Engineering Department, College of Engineering, University of Salahaddin-Erbil, Iraq. He received B.Sc. degree in Civil Engineering and M.Sc. in Sanitary Engineering from University of Salahaddin-Erbil, Iraq; Ph.D. in Environmental Engineering from Universiti Sains Malaysia (USM), Malaysia. He is editor and reviewer of some international journals. His area of specialization is Water Supply Engineering, Wastewater Engineering, Solid Waste Management, and Noise Pollution.

Professor M. Razip Selamat is at the School of Civil Engineering, Universiti Sains Malaysia. Dr. M. Razip received his PhD from University of Queensland, Australia.

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