METAL UPTAKE IN WATER SPINACH GROWN ON CONTAMINATED SOIL AMENDED WITH CHICKEN MANURE AND COCONUT TREE SAWDUST

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1 Environmentl Engineering nd Mngement Journl Septemer 214, Vol.13, No. 9, Gheorghe Ashi Tehnil University of Isi, Romni METAL UPTAKE IN WATER SPINACH GROWN ON CONTAMINATED SOIL AMENDED WITH CHICKEN MANURE AND COCONUT TREE SAWDUST Azln Kmri 1, Siti Njih Mohd Yusoff 1, Wiwid Prnt Putr 1, Che Fuzih Ishk 2, Norhyti Hshim 1, Azmi Mohmed 1, Esther Phillip 3 1 Universiti Pendidikn Sultn Idris, Fulty of Siene nd Mthemtis, Deprtment of Chemistry, 359, Tnjong Mlim, Perk, Mlysi 2 Universiti Putr Mlysi, Fulty of Agriulture, Deprtment of Lnd Mngement, 434, Serdng, Selngor, Mlysi 3 Mlysin Nuler Ageny, 43, Kjng, Selngor, Mlysi Astrt A pot experiment ws onduted to evlute the growth nd metl umultion in wter spinh (Ipomoe quti) estlished on 3-yer old tive firing rnge soil mended with hiken mnure (CM) nd oonut tree swdust (CTS) t pplition rtes of %, 1% nd 3% (w/w). Both mendments inresed iomss yield nd redued plnt metl uptke. The ioonentrtion ftor (BCF) nd trnsporttion ftor (TF) vlues of the metls were in the order of Zn > Cu > P. The mmonium ette extrtle metls in soil deresed signifintly (p <.5) following CM nd CTS tretments. It ws estimted tht the off-tke vlue of Zn ould e redued from 1.1 kg/h (zero tretment) to 6.6 kg/h (CM 3% w/w) nd 3.17 kg/h (CTS 3% w/w). No toxiity symptoms were oserved in wter spinh over the pot experiment. Therefore, hiken mnure nd oonut tree swdust re two promising gents for immoilizing hevy metls in ontminted lnd. Key words: hiken mnure, oonut tree swdust, ontminted soil, hevy metls, soil stiliztion, wter spinh Reeived: Mrh, 214; Revised finl: August, 214; Aepted: Septemer, Introdution Soil ontmintion y hevy metls is worldwide environmentl prolem. For exmple, in 25 metls were the min ontminnts ffeting 6 sites in Denmrk (Jensen et l., 29). Menwhile, Sun et l. (29) reported tht out 14,, h sites in Chin were ontminted y hevy metls, prtiulrly As nd Cd. Besides their nturl ourrene in the prent roks, speifi soures of hevy metls in soils re from nthropogeni tivities suh s mining, industry nd griulture (Swidis et l., 214; Kmri, 211). Ativities ssoited with firing rnges hve lso ontriuted to elevted levels of hevy metls in soil. Typil militry-grde ullets re minly omposed of P-lloy slugs enlosed with Cu-lloy jkets (Dermts et l., 24; Moon et l., 213). Metl prtiultes originting from multiple impts of ullet frgments during rnge opertions n e oxidized nd trnsformed into ompounds tht n e moilized (Chrstný et l., 21; Jorgensen nd Willems, 1997; Lndserger et l., 1999;). Moon et l. (213) mesured 11,885 mg/kg P in militry firing rnge soil olleted from Busn Metropolitn City, Kore. Topsoil smples olleted from firing rnge in New Mexio, USA ws reported to ontin 9,6 mg/kg P (Prr et l., 28). An nlysis on metl onentrtions in firing rnge soil of the Smll Arms Trining Are, Aiken, USA y Wilde et l. (25) found 3,282 mg/kg P nd 1,762 mg/kg Cu. Undoutedly, this senrio my pose gret risk to Author to whom ll orrespondene should e ddressed: e-mil: zlkm@yhoo.o.uk; Phone: ; Fx:

2 Kmri et l./environmentl Engineering nd Mngement Journl 13 (214), 9, the environment espeilly food hin nd wter supplies. A rnge of remedition strtegies exist for metl ontminted soils. Physil nd hemil tretments suh s exvtion nd lndfill, therml tretment, id lehing nd eletro-relmtion re effetive t lowering risk (EPA, 26; Jing et l., 29; Kshem et l., 21). However, these tehniques re expensive nd would e more idel for reltively smll ontminted sites (Bst nd MGowen, 24; Deel et l., 212). One promising remedition tehnique is the in situ stiliztion wherey n mendment is inorported into ontminted soil in order to immoilize hevy metls nd redue the dose(s) reeived y plnts (reeptors) without set-side effets (Hosseini et l., 213; Hrtley nd Lepp, 28). This tehnique dereses the hzrd potentil of the ontminted soil y onverting the ontminnts into their lest solule, moile or toxi form (Holm, 21; Krssim et l., 214). Amendments my ind, dsor or o-preipitte metl ontminnts, reduing metl moility nd vilility. Addition of orgni mtter to metl ontminted soil hs een employed to id their revegettion y improving fertility nd deresing the plnt-vilility of hevy metls (Aott et l., 21; Kumpiene et l., 27; vn Herwijnen et l., 27). Orgni mendments n derese hevy metls iovilility, shifting them from plnt-ville forms, tht is extrtle with wter or neutrl slts suh s lium hloride, to frtions ssoited with orgni mtter, rontes or metl oxides (Wlker et l., 24; Geeelen et l., 26). In ddition, the presene of funtionl groups on the surfe of orgni mendments would provide inding sites for hevy metls. Soil stiliztion using low-ost mterils hs een onsidered s ost-effetive tehnique for soil remedition. Numerous mendments hve een proposed nd tested for soil stiliztion, inluding griulturl nd industril y-produts. Lee et l. (213) studied metl immoiliztion in ontminted rie pddy soil using eggshell wste. The ddition of eggshell wstes t 5% (w/w) ws reported to redue the toxiity hrteristis lehing proedure (TCLP) of Cd nd P y 67.9 nd 93.2%, respetively. Moon et l. (213) reported tht mending firing rnge soil with iohr t n pplition rte of 2% (w/w) redued TCLP P lehility y 9%. Nwhukwu nd Pulford (29) mended ontminted soil with green wste ompost, pet nd wood rk t the rtes of 1%, 1% nd 2% (w/w). They reported tht P onentrtion in the shoots of perennil ryegrss (Lolium perenne) deresed signifintly with mendment tretments. Li et l. (212) treted ontminted soil using tnnery sludge hr. The immoiliztion effiienies of Zn, Cu nd Ni y tnnery sludge hr mendment were reported to e 84.2, 1. nd 4.%. A study y Fng et l. (212) hs shown tht phosphte rok tiling nd triple superphosphte fertilizer were le to redue CCl 2 -extrtle P nd Zn y % nd %, respetively. Cstldi et l. (29) studied metl uptke of pe nd whet ultivted on ontminted soil treted with red mud, zeolite nd lime t n pplition rte of 1% (w/w). The ddition of mendments ws found to redue Cd, P nd Zn umultion in plnt shoots nd roots. Intensive prodution of poultry met nd wood-sed produts hs generted lrge mount of wstes, espeilly mnure nd swdust. It is estimted tht out 44.4 million tons of poultry mnure ws produed in the USA in 28 (MDonld et l., 29). Menwhile, pproximtely 591,3 tons of roiler mnure wste is generted nnully in New Zelnd (Boln et l., 21). Both low-ost mterils re importnt soures of mironutrients for rop prodution nd orgni mtter for improving physil nd iologil fertility of soil (Dikiny nd Mufwnzl, 21; Stirling et l., 212). A numer of studies hve reported the potentil of mnure s n immoilizing gent for hevy metls in soil. Wlker et l. (24) reported tht mending metl ontminted soil with ow mnure inresed the growth of Chenopodium lum L. nd redued shoot onentrtions of Cu, Mn nd Zn. Sto et l. (21) evluted Cd phytovilility in soils s ffeted y pplition of hemil fertilizer nd three types of niml wste ompost (AWC), derived from ttle, swine nd poultry wstes. They reported tht the Cd onentrtion in spinh grown on soil mended with AWC ws 34-38% lower thn in plnts ultivted on hemil fertilizer-treted soil. The effetiveness of stiliztion strtegy depends on the nture of the ontminnts, physil nd hemil hrteristis of the mendment, nd type of soil. Hn et l. (212) studied Cd uptke y rie plnts grown on three different soils treted with pig mnure t the rtes of 1% nd 3% (w/w). Grin Cd onentrtions were reported to exeed the Chinese Ntionl Food Qulity Stndrd of.2 mg/kg most often on the lom, intermedite frequeny on the silt lom, nd lest often on the sndy lom. Mnure pplition in metl ontminted soil my not lwys redue metl vilility. Shw et l. (27) investigted hevy metl lehing from mine tilings mended with omposted ttle mnure, ged ttle mnure nd omposted yrd wste t n pplition rte of 9 tonne/h. They found tht ll mendments inresed the mount of hevy metl lehed from the tilings, s ompred to untreted mine tilings. As disussed y Nwhukwu nd Pulford (29), Lee et l. (213) nd Fng et l. (212), it is neessry to ssess the potentil of mendments for soil remedition in pot experiments, prior to full sle field studies. Suh informtion should e onsidered when pplying mendments on rel ontminted site, nd therefore, ostly future mistkes n e voided (Lee et l., 29; vn Herwijnen et l., 27). 222

3 Metl uptke in wter spinh grown on ontminted soil mended with hiken mnure nd oonut tree swdust The overll im of this work ws to evlute the potentil of hiken mnure nd oonut tree swdust s immoilizing gents for the remedition of metl ontminted soil. The ojetives of this work were: (1) to evlute the ility of hiken mnure nd oonut tree swdust to immoilize hevy metls in ontminted soil; nd (2) to study the effets of oth mendments on plnt growth nd plnt metl uptke. 2. Mterils nd methods 2.1. Soil nd mendments Soil smple ws olleted from 3-yer old tive firing rnge in Selngor, Mlysi. The soil smples were tken t the surfe lyer of up to 25 m depth using stinless steel trowel. The smples were ir-dried for 1 week, thoroughly mixed nd pssed through 2 mm mesh sieve. The soil texturl nlysis ws performed using the pipetting method s desried y Gee nd Buder (1986). The soil onsists of snd (79%), silt (16%) nd ly (5%). The soil ph ws mesured in deionised wter with soil:solution rtio of 1:2.5 using ph meter (Orion 2- Str, Thermo Sientifi, USA). The eletril ondutivity (EC) of soil ws mesured in soil nd wter suspension t rtio of 1:5 (w/v) using n EC meter (Orion 3-Str, Thermo Sientifi, USA). The Nelson-Sommers method (Nelson nd Sommers, 1996) ws used to determine the soil orgni mtter (OM), while the Kjeldhl distilltion proedure (Mremner, 1996) ws used to determine the totl N. The tion exhnge pity (CEC) ws determined using the mmonium-sturtion nd distilltion method (Sumner nd Miller, 1996). The totl Cu, P nd Zn onentrtions in the soil ws determined y qu regi extrtion. In ten replites, 9 ml of HCl (6. mol/l) nd 3 ml of HNO 3 (69%) were dded to.25 g of soil. The soilqu regi mixture ws left overnight for equilirtion. Soil smples nd lnks were digested t 125 ºC for 3 h. The digests were filtered through Whtmn No. 5 hrdened filter pper into 5 ml volumetri flsks nd mde up to volume with deionized wter. In ddition to the totl frtion, the ioville frtion of metls in soil ws lso determined using mmonium ette (1. mol/l, ph 7) t soil:extrtnt rtio of 1:1. The mixtures were gitted on n oritl shker for 1 h t room temperture nd were then filtered into smple ontiners. In this study, two orgni mendments nmely hiken mnure (CM) nd oonut tree swdust (CTS) were tested. Chiken mnure ws olleted from hiken frm in Klng, Selngor, while oonut tree swdust ws otined from sw-mill in Bnting, Selngor. The hiken mnure ws sun dried to kill ny germs ville whih ould hinder the growth of rop. The oonut tree swdust ws ir-dried for 1 week. The dried mendments were ground efore nlysis nd use. In order to mesure metl ontent in mendments, CM nd CTS were digested seprtely using 15 ml of nitri id (69%) t 11 ºC for 3 h. The hrteristis of the soil nd mendments re summrized in Tle 1. Tle 1. Chrteristis of the soil nd mendments used in the study Chrteristi Soil Chiken mnure Coonut tree swdust ph EC (ds/m) OM (%) Totl-N (g/kg) CEC (mol /kg) P (mg/kg) 2544 <.5 <.5 Cu (mg/kg) Zn (mg/kg) P (mg/kg) Cu (mg/kg) Zn (mg/kg) Totl frtion. Bioville frtion Pot experiments Pots with dimeter of 14. m nd height of 12. m were filled with 4 g of soil. CM nd CTS were dded seprtely to the soil t %, 1% nd 3% (w/w), in six replites. The soils were left to equilirte for two weeks. As the soil hs poor plnt nutrient ontent, ¼ strength Hoglnd s nutrient solution (Wtson et l., 23) ws dded to eh pot thrie week t pplition rte of 2 ml. The Hoglnd s nutrient solution ws pplied to the soils for two weeks only (equilirtion period). The ddition of nutrient solution ws disontinued when the pot experiment egn. After two weeks, eh pot ws tipped out nd remixed to ensure homogeneity nd to prevent the soil smples from eoming neroi. Wter spinh (Ipomoe quti) seed ws sown two weeks fter ddition of mendments. The pots were rrnged in rndomized lok design. The wter ontent of the soils ws djusted to otin 7% of the wter holding pity y dding deionized wter dily, voiding prolonged wter logging. Plnts were llowed to grow under nturl lighting nd temperture for 8 weeks. Men dily temperture nd humidity were monitored with digitl thermometer. At the end of the pot experiment, the soil ph nd mmonium ette extrtle metl ontent in the soil were determined, s previously desried Hrvest nd digestion of plnt tissue The plnts were hrvested fter 8 weeks of growth. The eril prts were ut t 1. m ove the soil surfe to void ontmintion y soil using pir of sissors, whih ws wiped fter eh use. Roots were refully extrted from the soil nd 2221

4 Kmri et l./environmentl Engineering nd Mngement Journl 13 (214), 9, wshed thoroughly with deionized wter to remove soil prtiles. Plnt tissues were dried in n oven t 7ºC for 48 h. After two dys, the dry weight of plnt tissue ws mesured. The dried shoots nd roots were milled seprtely using grinder. In triplite,.25 g milled plnt tissue ws shed in furne t 45 ºC for 3 h. In 5 ml tll form eker, 12 ml of HNO 3 (69%) ws dded to the shed smple. The plnt smple ws left overnight in the fume upord for equilirtion. Smples nd lnks were then digested on hot plte t 11 ºC for 3 h, llowed to ool nd then filtered through Whtmn No. 5 hrdened filter pper into 25 ml volumetri flsks. The solutions were mde up to volume using deionized wter. A Perkin-Elmer AAnlyst 4 Atomi Asorption Spetrometer ws used to mesure the onentrtion of hevy metls in plnt tissue digests nd soil extrts Stndrd nd ertified referene mterils Stndrd referene plnt mterils (SRM 1573 Tomto Leves Ntionl Institute of Stndrds & Tehnology, USA, nd SRM 1575 Pine Needles Ntionl Bureu of Stndrds, USA) nd ertified referene soil mteril (LGC 6135 Hkney Brik Works Soil Lortory of the Government Chemist, UK) were used to verify the ury of metl determintion. Referene mterils were treted nd nlyzed using the sme proedures pplied for plnt tissue nd soil smples. The reovery rtes were within 92-15% for soil nd 88-94% for plnt tissue, respetively (Tle 2). Tle 2. Metl onentrtions in referene mterils Referene P Cu Zn mteril LGC 6135 Certified 391 ± ± ± 5 Mesured 358 ± 6 3 ± ± 7 Reovery (%) SRM 1573 Certified ± ±.1 Mesured ± ±.6 Reovery (%) SRM 1575 Certified 1.8 ±.5-3. ±.3 Mesured 9.5 ± ±.8 Reovery (%) Vlues represent men of three replites ± stndrd devition Sttistil nlysis All sttistil nlyses were performed using Minit 15 Sttistil Softwre (Minit In., PA, USA). The dt were nlyzed using the generl liner model of one-wy nlysis of vrine (ANOVA), followed y Tukey s test t signifine level of p =.5 to determine lest signifint differene (LSD) for the omprison of mens. Correltion ws y Person s oeffiients t p < Results nd disussion 3.1. Plnt growth nd iomss yield In this study, wter spinh seeds were lso plnted on ompost nd reeived no mendment, in six replites. This enled diret omprison in terms of growth performne of wter spinh grown on untreted ontminted soil (zero tretment) nd unontminted soil (ompost). The wter spinh seeds germinted three dys fter sowing nd no ovious differene in plnt growth ws oserved up to two weeks of the pot experiment. Plnts grown on ompost were oserved to e helthier thn plnts ultivted on untreted ontminted soil. CM nd CTS tretments hve resulted in helthy pperne on the wter spinh leves, wherey the leves were oth longer nd greener s ompred to plnts grown on untreted ontminted soil. The growth progress of plnts grown on zero tretment soil ws found to e slower thn plnts grown on soil reeived mendments. However, no toxiity symptom suh s reddish or urnt pperne ws pprent on leves of untreted wter spinh. This suggests tht wter spinh is roust plnt speies nd hs gret tolerne to high metl onentrtions. Tle 3 presents the dry iomss yield of wter spinh fter 8 weeks of growth. Tle 3. Biomss yield of wter spinh Dry weight (g/pot) Tretment Shoot yield Root yield Compost* Zero CM 1% CM 3% 1.75 d 3.47 d CTS 1% CTS 3% LSD *Plnts grown on ompost only (unontminted soil). Vlues represent men of 6 replites. Letters,, nd d show the signifint differenes etween the soil tretments, where letter represents the lowest men. Different letters indite signifint sttistil differenes (Tukey s test t p <.5). As expeted ddition of CM nd CTS to ontminted soil inresed iomss yield. From Tle 3, it is ler tht the shoot nd root yields inresed with the rtes of mendment pplition. A pronouned effet ws otined for CM tretment t 3% (w/w), of whih the shoot yield for this tretment ws found to e higher thn zero tretment y ftor of 3.2. CM 1% nd CTS 3% (w/w) tretments gve lmost similr shoot yield, with % inrement in iomss prodution. The highest perentge of inrement in root yield ws hieved with CM 3% (w/w) tretment, followed y pplition of CM t 1% (w/w). Although there ws n inrese in the root yield following pplition of the CTS, sttistil nlysis reveled no signifint differene ws otined etween the CTS 1% nd CTS 3% (w/w) tretments. 2222

5 Metl uptke in wter spinh grown on ontminted soil mended with hiken mnure nd oonut tree swdust CM nd CTS were enefiil s growing medi through improvement of soil fertility nd provision of plnt nutrient. CM mendment resulted in higher iomss yield thn CTS. This n e explined y the ft tht CM ontins higher nutrient nd orgni mtter thn CTS. As presented in Tle 1, the nitrogen ontent mesured in CM ws 62.7 g/kg menwhile 25.5 g/kg nitrogen ws determined in CTS. The orgni mtter ontent in CM nd CTS ws estimted s 87.6% nd 72.9%, respetively. Lower yields of shoot nd root otined for zero tretment plnts n e ttriuted to metl toxiity. In mny ses iomss yield hs signifint liner response to pplition rtes of mendment. Wlker et l. (24) otined signifint liner response for dry mtter prodution of C. lum to inorportion rtes of ow mnure dded to metl ontminted soil. Li et l. (28) reported tht mending metl ontminted pddy soil with higher pplition rte of pig mnure resulted in greter yield of rie grin. A different oservtion ws reported y Chiu et l. (26), who studied the effet of pig mnure ompost nd sewge sludge mendment on the growth of ommon reed plnt ultivted on P/Zn mine tilings. They noted tht pplition of mnure ompost t 5% nd 1% (w/w) did not signifintly ffet the shoot yield of ommon reed, wherey the dry mtter prodution ws found to e rther similr to tht of plnts grown on untreted mine tilings Metl onentrtion in plnt tissue The onentrtions of Zn, P nd Cu in plnt tissues fter 8 weeks of growth+ re given in Figs. 1 nd 2. It is pprent tht CM nd CTS tretments redued metl onentrtions in shoots nd roots of wter spinh. In generl, shoot nd root metl onentrtions deresed with the rtes of mendments pplition. Mrked redutions in metl onentrtions were hieved following CTS tretment t 3.% (w/w). From Fig. 1, lthough CM tretments redued Zn shoot onentrtion, there ws no signifint differene etween 1% nd 3% (w/w) tretments. Applition of CM t 1% (w/w) deresed P shoot onentrtion y only 9%, s ompred to plnts grown on untreted ontminted soil. Menwhile, CM 3% nd CTS 1% (w/w) tretments were found to hve similr effets in reduing P shoot onentrtions. CM 1% (w/w) hd the lest immoilizing effet wherey the onentrtion of Cu in wter spinh shoots ws redued from 1.5 mg/kg (zero tretment) to 9.9 mg/kg. In the se of metl root onentrtions (Fig. 2), no signifint derese ws oserved fter mending ontminted soil with CM t 1% (w/w). A signifint derese (p <.5) in metl root onentrtions ws otined for CM 3%, CTS 1% nd CTS 3% (w/w) tretments. The ddition of CM inresed soil ph from 4.91 to 7.73, reduing metl vilility for metl uptke in wter spinh tissues. The ph vlue of CM ws determined s 8.75 (Tle 1). A similr oservtion in soil ph inrese following CM pplition ws lso reported y Pitmn nd Singh (1993) nd Hue (1992). The inrese in soil ph n e ttriuted to ion exhnge mehnisms whih our when terminl OH - of Al or Fe 2+ hydroxyl oxides re repled y orgni nions whih re deomposed produts of the CM suh s mlte, itrte nd trtrte (Dikiny nd Mufwnzl, 21; Hue, 1992). In ddition, the presene of si tions in the CM relesed upon miroil deroxyltion n lso inrese soil ph (Boln et l., 21; Pitmn nd Singh, 1993). The effet of CM pplition on soil ph is gretly influened y the nture of the soil. For exmple, Dikiny nd Mufwnzl (21) noted tht the ddition of CM did not hnge the ph of the Luvi Clisol, menwhile sustntil inrese ws oserved for Ferrli Arenosol nd Verti Luvisol. It is known tht orgni mtter onsists of humi sustnes, whih re le to form stle metl heltes (Wlker et l., 24). This redues the soluility of hevy metls in soil. As disussed y Kumpiene et l. (28) nd Moon et l. (213), mendments with funtionl groups re prtiulrly effetive t immoilizing metl due to their ility to ind or omplex metl. In order to identify the presene of ertin funtionl groups nd hemil onds on CTS, s well s to understnd the nture of metl inding y CTS, Fourier Trnsform Infrred (FTIR) nlysis ws rried out. Zn in shoots (mg/kg) Cu in shoots (mg/kg) P in shoots (mg/kg) d Tretment Fig. 1. Metl onentrtions in wter spinh shoots fter 8 weeks of growth. Vlues represent men of 18 replites ± stndrd devition. Different letters indite signifint sttistil differenes (Tukey s test t p <.5). CM 1: Chiken mnure 1% (w/w), CM 3: Chiken mnure 3% (w/w), CTS 1: Coonut tree swdust 1% (w/w), nd CTS 3: Coonut tree swdust 3% (w/w). 2223

6 Kmri et l./environmentl Engineering nd Mngement Journl 13 (214), 9, Zn in roots (mg/kg) It is evident from FTIR nlysis tht OH nd NH 2 funtionl groups re ville on the surfe of CTS. FTIR spetr suggest tht metl inding ffets ll hemil onds ssoited with O nd N toms. Hene, it is resonle to speulte tht O nd N toms re indeed the min inding sites for metl on CTS, wherey oth toms n ind metl ions through omplextion. P in roots (mg/kg) Cu in roots (mg/kg) Fig. 3. FTIR spetr of CTS efore nd fter metl inding Tretment Fig. 2. Metl onentrtions in wter spinh roots fter 8 weeks of growth. Vlues represent men of 18 replites ± stndrd devition. Different letters indite signifint sttistil differenes (Tukey s test t p <.5). CM 1: Chiken mnure 1% (w/w), CM 3: Chiken mnure 3% (w/w), CTS 1: Coonut tree swdust 1% (w/w), nd CTS 3: Coonut tree swdust 3% (w/w) FTIR nlysis ws performed on Thermo Niolet 67 FTIR Spetrometer using the KBr dis tehnique. A rod sorption nd oserved t 3327 m -1 in Fig. 3 orresponds to the strething virtion of O-H. The FTIR spetrum of CTS exhiits hrteristi frequenies of C=O strething virtion for ronyl group t 1718 m -1. The NH 2 sissoring virtion ws oserved t 1596 nd 156 m -1. Aording to Crews et l. (1998), the sorption nd t 1232 m -1 orresponds to the C-N strething virtion. The C-O strething virtion for primry lohol groups ppered t 129 m -1 (Willims nd Fleming, 1995). Intertion of CTS with hevy metls led to shift in wve numer from 3327 m -1 to 3271 m -1 for O-H strething virtion nd from 1718 m -1 to 1723 m -1 for C=O strething virtion. A signifint hnge in sorption intensity of C=O, NH 2, C-N nd C-O strethes ws oserved. It is ler tht pplition of mendments to ontminted soil my influene the ehvior of plnt in umulting metls. This ehvior is minly ffeted y severl ftors suh s the nture of the soil, mendment nd metl ontminnt, pplition rte of mendment nd plnt speies. Correltions etween metl onentrtions in soil nd metl onentrtions in plnt tissues were mde using two extrtnts, nmely EDTA nd mmonium ette (NH 4 OA) (Tle 4). It ws found tht mmonium ette gve signifint orreltions etween metl onentrtions in soil nd metl onentrtion in plnt tissues. In ontrst, EDTA exhiited poor orreltions. The poor orreltion etween EDTA extrtle metl onentrtions nd plnt tissue metl onentrtions my e euse EDTA is good extrtnt for metl ssoited with orgni mtter, whih my not e ville for uptke y plnts (Nwhukwu nd Pulford, 29) Bioonentrtion ftor (BCF) nd trnslotion ftor (TF) The ioonentrtion ftor is defined s the rtio of metl onentrtion in plnt shoots to metl onentrtion in soil (MGrth nd Zho, 23). Tle 4. Correltions etween metl onentrtions in soil nd metl onentrtions in plnt tissue of wter spinh Shoot tissue Root tissue Extrtnt Metl Correltion p-vlue Correltion p-vlue oeffiient oeffiient EDTA P.11 NS -.48 NS NH 4 OA Cu.26 NS NS Zn.39 NS -.64 NS P.385.3*.571.2* Cu.514.2*.328.1* Zn.746.*.62.4* n = 65, NS: Not signifine, Person s orreltion oeffiient nd signifine t p <.5). 2224

7 Metl uptke in wter spinh grown on ontminted soil mended with hiken mnure nd oonut tree swdust As disussed y Yoon et l. (26), BCF is mesure of the ility of plnt to umulte metls from soil. Plnts with BCF vlue of greter thn 1 re suitle for phytoextrtion (Ruiz et l., 29). In this study, the influene of CM nd CTS tretments on BCF vlues of the metls ws determined nd the vlues re given in Tle 5. Tle 5. BCF vlues for P, Cu nd Zn Tretment BCF P Cu Zn Zero.19 d.22 d 1.15 d CM 1%.15 d CM 3% CTS 1% CTS 3% LSD Vlues represent men of 18 replites. Letters,, nd d show the signifint differenes etween the soil tretments, where letter represents the lowest men. Different letters indite signifint sttistil differenes (Tukey s test t p <.5). The BCF vlue of Zn for plnts ultivted on zero tretment soil ws lulted s 1.15, suggesting tht wter spinh hs gret potentil for phytoextrtion of Zn from ontminted soil. CM 1% nd 3% (w/w) tretments hd similr effets on Zn umultion, wherey the BCF vlue of Zn ws found to derese from 1.15 (zero tretment) to.63 (CM 1%) nd.58 (CM 3%), respetively. The pplition of CM t the rte of 1% (w/w) did not signifintly (p <.5) redue P uptke in wter spinh. Overll, the ility of wter spinh to tke up metls from ontminted soil deresed with the ddition of mendments. The derese in metl umultion n e ttriuted to metl inding to funtionl groups of mendments nd redution in metl vilility in soil, s disussed in the preeding setion. The onentrtion of metl in plnt shoot nd soil gretly ffets the BCF vlue. In this study, the BCF vlues of Zn were found to e higher thn the BCF vlues omputed for P nd Cu (Tle 5). This n e explined y the ft tht more Zn ws mesured in the plnt shoots s ompred to P nd Cu (Fig. 1). In ddition, the totl onentrtion of Zn in the ontminted soil (318 mg/kg) ws lower thn the 2544 mg/kg mesured for P. The trnslotion ftor is defined s the rtio of metl onentrtion in plnt shoots to metl onentrtion in roots (Ruiz et l., 29), nd is mesure of the ility of plnt to trnslote metls from the roots to the shoots (Yoon et l., 26). From phytoremedition viewpoint, n effiient phytoextrtion strtegy should hve high TF vlues euse metls must e in the eril prts of the plnts so tht they n e removed t hrvest (Pulford nd Dikinson, 26; Vmerli et l., 21). The effets of CM nd CTS tretments on the TF vlues of the metls re given in Tle 6. Wter spinh grown on ontminted soil reeived no mendment gve TF vlue of 1.9 for Zn. Aording to Bker nd Brooks (1989) nd Cunninghm et l. (1995), plnts with TF vlue of 1 re effiient in trnsporting metls from the root tissue to the shoot tissue. Therefore, phytoextrtion of Zn using wter spinh is fesile. However, mending ontminted soil with CM nd CTS redued the trnslotion of metls in wter spinh, espeilly with CTS t pplition rte of 3% (w/w). Following CTS 3% (w/w) tretment, the BCF vlue of Zn deresed signifintly (p <.5) from 1.9 (zero tretment) to.14. There ws no gret differene in the TF vlue of P for plnts grown soil treted with CM 3% nd CTS 1 % (w/w). Tretment Tle 6. TF vlues for P, Cu nd Zn TF P Cu Zn Zero.14 d.25 d 1.9 d CM 1% CM 3% CTS 1% CTS 3% LSD Vlues represent men of 18 replites. Letters,, nd d show the signifint differenes etween the soil tretments, where letter represents the lowest men. Different letters indite signifint sttistil differenes (Tukey s test t p <.5). In generl, the BCF nd TF vlues of P, Cu nd Zn showed similr trend following the immoiliztion tretments. This suggests tht oth mendments hd similr effet on metl umultion nd trnsport in wter spinh tissue. Bsed on the BCF nd TF vlues, metl uptke in wter spinh tissue is in the order of Zn > Cu > P. Zn nd Cu re essentil mironutrients for plnt growth. Therefore, the trnsport nd umultion of Zn nd C were found to e higher thn P. Unlike Zn nd Cu, P normlly uses toxi effet to plnts (Kshem et l., 21; Vmerli et l., 21). The toxi effet of P on hlorophyll synthesis, photosyntheti tivity nd ntioxidnt enzymes hs een disussed y Kim et l. (23). Overll, the trend otined for BCF nd TF vlues orroortes the trend oserved for metl onentrtion in plnt roots (Fig. 2). P ws the metl most mesured in plnt roots nd the lest trnsported from the root to the shoot of wter spinh. There re two min mehnisms holding P in plnt roots, nmely inding of P to ion exhnge sites on the root ell wlls nd extrellulr preipittion (Jrvis nd Leung, 22). It is known tht plnts hve different ilities to tke up nd trnslote hevy metls in their tissues. For exmple, mize nd sunflower hd preferene of Zn > Cu > P (Ruiz et l., 29). Menwhile, Yoon et l. (26) reported the order of P > Cu > Zn for 2225

8 Kmri et l./environmentl Engineering nd Mngement Journl 13 (214), 9, Cyperus esulentus nd Cu > Zn > P for Ruus frutiosus Off-tke vlues The effet of the pplition of CM nd CTS on metl uptke in wter spinh ws further evluted in terms of off-tke vlue. The off-tke vlue onsiders oth metl onentrtion in plnt tissues nd iomss yield, nd hs een regrded s n importnt prmeter in evluting the effetiveness of mendments for metl immoiliztion (Kmri, 211). The off-tke vlue ws lulted per pot (mg/pot) nd per hetre (kg/h). The estimtion in kg/h ws sed on onversion ftor of pot re to hetre. The mount of P, Cu nd Zn removed y wter spinh from ontminted soil is given in Tle 7. From Tle 7, it is oserved tht pplition of CM nd CTS redue the removl of P, Cu nd Zn y wter spinh. The influene of mendments ws less pprent for P uptke euse P ws the metl lest extrted y plnts. At the end of the pot experiment, it is estimted tht.7 mg/pot of P,.36 mg/pot of Cu nd 3.85 mg/pot of Zn were removed from the untreted ontminted soil. Amending ontminted soil with CM nd CTS t the rte of 3% (w/w) deresed the off-tke vlue of Zn from 1.1 kg/h (zero tretment) to 6.6 nd 3.17 kg/h, respetively. The redution in the off-tke vlues of metls n e ttriuted to the immoiliztion effet of the hiken mnure nd oonut tree swdust. l., 21; Pulford nd Dikinson, 26). Therefore, the effet of the pplition of CM nd CTS on ioville frtion of P, Cu nd Zn in ontminted soil ws investigted using mmonium ette s extrtnt. The mmonium ette extrtle metls in soils fter 8 weeks of the pot experiment re given in Tle 8. Amending ontminted soil with CM nd CTS deresed the iovilility of metls signifintly (p <.5), prtiulrly t pplition rte of 3% (w/w). A lesser effet in reduing the metl iovilility ws otined when mendments were pplied t 1% (w/w). The derese in ioville frtion of hevy metls fter the pot experiment n e ttriuted to immoiliztion effet of the mendments nd uptke y plnts. Sine plnts were wtered with deionized wter dily, metls my hve lehed from the soil to the pot suer. Therefore, lehing my hve lso ontriuted to the derese in metl onentrtion of soil. Tle 8 lso presents the perentge derese in the mmonium ette extrtle metls in the soil fter the pot experiment. The untreted ontminted soils (ontrols) show derese in ioville frtion of hevy metls. This ould e due to the hnges in the hemil form of the metl tht re not relted to the mendments during the pot experiment. It is pprent tht CTS 3% (w/w) ws the est tretment in lowering ioville frtion of metls, menwhile CM 1% (w/w) ws the lest effetive tretment in immoilizing metls. 4. Conlusions 3.5. Bioville frtion of metls Results from this study highlight the potentil The uptke of hevy metls y plnts is of hiken mnure nd oonut tree swdust s minly influened y their ioville frtion immoilizing gents for the remedition of metl rther thn y the totl mount in soil ( Vmerli et ontminted lnd. Pot experiment however is only one spet of suh utiliztion. Tle 7. Removl of P, Cu nd Zn from ontminted soil using wter spinh Tretment Off-tke (mg/pot) Off-tke (kg/h) P Cu Zn P Cu Zn Zero CM 1% CM 3% CTS 1% CTS 3% Tle 8. Ammonium ette extrtle metls in soils Tretment Conentrtion (mg/kg) Derese (%) P Cu Zn P Cu Zn Zero d 129 d CM 1% d CM 3% CTS 1% CTS 3% LSD Vlues represent men of 18 replites. Letters,, nd d show the signifint differenes etween the soil tretments, where letter represents the lowest men. Different letters indite signifint sttistil differenes (Tukey s test t p <.5). 2226

9 Metl uptke in wter spinh grown on ontminted soil mended with hiken mnure nd oonut tree swdust The effetiveness of hiken mnure nd oonut tree swdust s soil mendments rely on their stility in the soil-wter environment. It is neessry to study the iodegrdtion of oth mendments nd their effet on metl iovilility. Aknowledgments The uthors thnk Ministry of Edution Mlysi nd Universiti Pendidikn Sultn Idris Mlysi for providing reserh grnts (RACE nd GPU ). We thnk Mr. Mohd Zurin, Mr. Mohd Hshimi, Miss Siti Nor Frin nd Mrs. John for their ssistne. 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