Effects of EDTA on phytoextraction of heavy metals (Zn, Mn and Pb) from sludge-amended soil with Brassica napus

Bioresource Technology 101 (2010) 3978–3983

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Effects of EDTA on phytoextraction of heavy metals (Zn, Mn and Pb) from sludge-amended soil with Brassica napus Hanen Zaier a,*, Tahar Ghnaya a, Kilani Ben Rejeb a, Abdelbasset Lakhdar a, Salwa Rejeb b, Fatima Jemal c a

Laboratoire d’Adaptation des plantes aux Stress Abiotiques, Centre de Biotechnologie, Technopole Borj-Cédria, BP 901, 2050 Hammam-lif, Tunisia Laboratoire de Physiologie Végétale de l’INRGREF, Tunisia c Unité de Recherche Nutrition et Métabolismes Azotés et Protéines des Stress, Faculté des Sciences de Tunis, Tunisia b

a r t i c l e

i n f o

Article history: Received 7 August 2009 Received in revised form 27 October 2009 Accepted 11 January 2010 Available online 2 February 2010 Keywords: Chelator Phytoremediation Plant biomass Pollution Sewage sludge

a b s t r a c t Sludge application is a reliable practice to ameliorate soil fertility. However, repetitive sludge addition represents a potential soil contamination source with heavy metals, which must be extracted. The aim of this study was to evaluate the capacity of Brassica napus to remove metals from soils amended with sludge, and to study the effect of EDTA on this process. Seedlings were cultivated in presence of sludge combined or not with EDTA. Results showed that sludge ameliorate significantly biomass production. This effect was accompanied with an increase in Pb, Zn and Mn shoot concentrations. EDTA application does not affect significantly plant growth. However, this chelator enhances shoot metals accumulation. It’s therefore concluded that sludge has a beneficial effect on soil fertility, B. napus can be used for the decontamination of affected soils and that the EDTA addition increases the ability of B. napus to accumulate heavy metals. Published by Elsevier Ltd.

1. Introduction In order to ameliorate the physicochemical proprieties of soil, the use of sewage sludge in agriculture is a worldwide practice (Gupta and Sinha, 2007; Singh and Agrawal, 2008). Since sludge is a result of wastewater treatment, which is generally bulky with high moisture and from highly organic and mineral, these fertilizers are rich with organic matter and mineral elements (N, P, K, Ca) essential for plant growth (Singh and Sinha, 2004a,b; Tsadilas et al., 2005). However, the characterization of sewage sludge metal is an important requirement prior to sludge disposal to farmland because there is a major risk of toxic elements accumulation in soil (Gupta and Sinha, 2006, 2007). Sewage sludge may contain some trace elements, such as heavy metals (Cd, Mn, Zn, Pb. . .) which are toxic for plant, animal and human (Renoux et al., 2007). Recent study demonstrated that the extensive addition of sludge to agricultural soils lead to an excessive accumulation of trace metals which are not biodegradable and tend to accumulate in soils, waters and crop plants or transferred to human thought contaminated food (Singh and Agrawal, 2007). In fact, very high amounts of Fe, Cu and Zn were observed in maize and barley grown on sludgeamended soil than those grown on the unamended ones (Hernandez et al., 1991). Because of the potential toxicity and the persis-

* Corresponding author. E-mail address: [email protected] (H. Zaier). 0960-8524/$ - see front matter Published by Elsevier Ltd. doi:10.1016/j.biortech.2010.01.035

tence of heavy metals, the clean up of agricultural amended soils is one of the most difficult tasks for environmental engineering. Physicochemical methods for heavy metals removal from contaminated soils are expensive and soil disturbing by intensive labor (Gardea-Torresdey et al., 2005). Recently, the use of plants to remediate polluted soils, or phytoremediation, has appeared as an alternative more reliable (Arthur et al., 2005; Cho et al., 2009). This emerging biological approach, based on the capability of some plant species to take up and to concentrate pollutants in their shoots, is often simpler in design and cheaper to implement (Blaylock et al., 1997; McGrath et al., 2002; Petra et al., 2009). The use of hyperaccumulators species in continuous phytoextraction process is limited by the low bioavailability of theses pollutants to the root uptake (Salt et al., 1998; Peer et al., 2005). On the other hand, the majority of hyperaccumulators present a slow growth rate leading to a low annual biomass (Brooks, 1994; Peer et al., 2005). More recently, it was demonstrated that the application of mobilizing/chelating agents, such as ethylene diaminetetraacetic acid (EDTA) to the soil is a reliable practice to increase plant metal bioavailability, uptake and shoot accumulation (Evangelou et al., 2007; Meers et al., 2008; Luo et al., 2008; Petra et al., 2009). Such chelator presents a strong affinity for different heavy metals cations (Cooper et al., 1999; Römkens et al., 2002; Shen et al., 2002; Meers et al., 2005; Lotte et al., 2007) and is readily translocated from the roots to the shoots as a metal–chelate complex (Lombi et al., 2001; Collins et al., 2002; Wenger et al., 2003; Tandy et al., 2006). Hence, chelator enhance in the same time the uptake

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and the translocation of heavy metals (Wu et al.,2004; Tandy et al., 2006), and it can also reduce the toxicity of free metal-cations in the photosynthetic organs by its complexation (Vassil et al., 1998; Nowack, 2002; Greˇman et al., 2003; Hernandez-Allica et al., 2003). Among other characteristics, for chemically-induced phytoextraction, ideally plants should be fast-growing, have the capability to tolerate and accumulate high concentrations of the metal in their shoots (McGrath et al., 2002). Brassica napus, belonging to Brassicacea family, is recognized as fast-growing metal-accumulator species, constituting a good candidate for induced phytoextraction (Veerle Grispen et al., 2006). On the other hand, the choice of the chelator could determinate the success of this process. Hence scientists used currently EDTA as the adequate chelator for the induction of heavy metals uptake and translocation (Shen et al., 2002; Van Engelen et al., 2007; Quartacci et al., 2006, 2007). The present study was carried out to assess the effectiveness of B. napus to absorb and accumulate heavy metals (Zn, Mn and Pb) in their shoots from sludge-amended soils and to evaluate the possible effect of EDTA application on this process.

Table 1 Physical and chemical characteristics of sludge and soil used in the experiment. All these values are the means of four replicates. Nd no determined.

2. Methods

Available phosphorus was determined according to Egner et al. (1960). Soil and sludge samples were digested with a mixture of HCl/HNO3 (McGrath and Cunliffe, 1985) and total concentrations of heavy metals in soils and sludge were determined using an Atomic Absorption Spectrometer (PERKIN ELMER Analyst 300). K+ and Na+ contents were determined in the same homogenate by flame spectrometry (Corning photometer).

2.1. Field experiment design Our experimentation was carried out under natural conditions experimental field located in Oued souhil in the north of Tunisia (state of Nabeul) 60 km north-east of Tunis. The site was characterized by a semi-arid climate with a mean annual precipitation of 456 mm (mainly during the winter) and an average temperature of 19.4 °C. The soil had clayey sandy texture characterized by low level of organic matter and the electric conductivity about 1 mmhos/cm in the superficial horizons (0–30 cm) (Table 1). The experimental field covered a total surface area of 196 m2 (14  14 m). The total area was divided into 16 elementary plots of 4 m2 (2  2 m) separated by a neutral zone of 2 m. The experiment was carried out using tow repetitive plot for each treatment, arranged randomly. The eight treatments were: 1: 2: 3: 4:

0 t/ha sludge (control) 20 t/ha sludge 40 t/ha sludge 80 t/ha sludge

5: 6: 7: 8:

0 t/ha sludge + EDTA 20 t/ha sludge + EDTA 40 t/ha sludge + EDTA 80 t/ha sludge + EDTA

The sludge was mixed thoroughly with the soil and applied by hand digging 15 day before seeds sowing at depth of 0–30 cm. After sludge application seeds of B. napus were sown at the density of 28 plants/4 m2 (56 plants per treatment). Eight weeks later, EDTA was added (3 g EDTA/kg dry soil) in the treatment 5, 6, 7 and 8. During the period of the treatment, irrigation was achieved every 3 days with tap water. Sewage sludge used in this study was obtained from a station of waste water treatments of Nabeul. It was previously aerobically stabilized and desiccated. The physico-chemical characteristics of used sludge are summarized in Table 1. 2.2. Soil and sludge chemical analysis The chemical properties of sewage sludge and soil studied were determined by standard methods. The pH was measured potentiometrically in 1 M KCl after 24 h in the water/soil ratio of 5. Electrical conductivity (EC) and the percentage of organic carbon were determined with the method of Kalra and Maynard (1991). The total nitrogen (Nt) was determined by the Kjeldahl’s method.

Parameters

Sewage sludge

Soil

pH Relative humidity (%) CE (mmhos/cm) Organic carbon (g kg1) Organic matter (g kg1) Total nitrogen (g kg1) C/N Phosphorus (mg kg1) K+ (g kg1) Ca2+ (g kg1) Mg2+ (g kg1) Na+ (g kg1) Mn (mg kg1) Zn (mg kg1) Pb (mg kg1) Cu (mg kg1) Co (mg kg1) Cd (mg kg1)

4.2 ± 0.21 1.8 ± 0.18 3 ± 0.3 367 ± 18.35 632.7 ± 31.6 30.5 ± 1.25 12.03 ± 2.40 29.7 ± 1.48 1.30 ± 0.26 16.86 ± 3.37 3.16 ± 0.3 0.85 ± 0.17 234 ± 11.17 515 ± 25.7 301 ± 15.05 155 ± 7.75 10 ± 0.5 2 ± 0.4

7.3 ± 1.46 1.5 ± 0.15 1 ± 0.4 9.8 ± 1.22 16.8 ± 2.1 1 ± 0.4 9.8 ± 1.34 8.50 ± 1.06 0.19 ± 0.023 3.74 ± 0.53 0.49 ± 0.024 0.28 ± 0.04 168 ± 9.33 59 ± 11.8 98 ± 12.25 20 ± 3.33 3 ± 0.56 Nd

2.3. Measured parameters During the vegetative stage of plant development, five periodic harvests were performed. The first one was applied the day of the application of EDTA (8 weeks after seeds sowing). The other harvests were conducted every 2 weeks after the application of EDTA. At the harvests, six uniforms plants for each treatment were considered and divided into shoots and roots. Shoots were successively rinsed three times in cold water and blotted between two layers of filter-paper. Roots were dipped for 5 min in a 0.01 M HCl cold solution to eliminate any external heavy metal adsorbed at the root surface, rinsed three times with cold distilled water (Aldrich et al., 2003) and blotted with filter-paper. The fresh weight was measured immediately, and the dry weight after 48 h of desiccation in an oven at 60 °C. The tolerance index (Ti) is defined according to Cottenie et al. (1983) by the relationship between the growths on a soil enriched compared to the growth on a control soil.

Ti ¼

Growth on a soil enriched Growth on a control soil

The value of Ti is equal to 1 when there is no influence of treatment on the growth, higher than 1 when there is a favorable effect of sludge on the growth and lower than 1 when the growth is affected negatively by the treatment. 2.4. Cations assay in plant Desiccated plant samples were grounded to a fine powder using a porcelain mortar and a pestle. Shoots and roots grinded samples (100 mg) were digested in 4/1 (v/v) HNO3/HClO4 (20 ml) mixture at temperature of 100 °C. After total evaporation, 30 ml of HNO3 0.5% were added to the remained residue. Then, heavy metals (Pb2+, Mn2+, Zn2+) and nutrients (Ca2+and Mg2+) concentrations were determined by atomic absorption spectrometry (PERKIN ELMER Analyst 300).

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2.5. Statistical analysis Analyses of variance (ANOVA) with orthogonal contrasts and mean-comparison procedures were used to detect differences between treatments. Mean-separation procedures were carried out using the multiple range tests with Fisher’s least significant difference (LSD) (P < 0.05). 3. Results

Table 2 Plants biomass production and tolerance index in Brassica napus cultivated during 4 months in the presence of different sludge’s amounts and in the absence of EDTA. Data are the means of six replicates. Mean values followed by the same letter are not significantly different at p = 0.05. Sludge doses (t/ha)

Whole plants dry weight (g)

Tolerance index (TI)

0 20 40 80

38.65 ± 0.60 a 55.65 ± 0.14 b 78.38 ± 0.22 c 116.24 ± 0.16 d

1.44 ± 0.02 2.04 ± 0.03 3.02 ± 0.04

3.1. Effect of sludge on the biomass production of B. napus Results showed that sludge does not affect significantly plant biomass production in B. napus, during the two first months after sowing. Hence, in the first harvest plant biomass were similar in the presence of different sludge doses. However, at the final harvest, sewage sludge stimulated significantly the biomass production of B. napus, compared to control (Fig. 1). This effect increased with the increasing sludge amounts and the duration of culture. Thus, 16 weeks after sowing, biomasses measured in plants cultivated in the presence of different sludge doses, as compared to control ones, represented 150%, 200% and 300%, respectively, at 20 t/ha, 40 t/ha and 80 t/ha. According to Table 2, we notice that the index of tolerance for the three treatments related to the sludge doses is higher than 1 and reaches 3 for the 80 t/ha treatment. These data confirm the beneficial effect of the sludge on the growth of B. napus probably due to its fertilizing effect.

150

A

-EDTA +EDTA

100

+ EDTA

50

a

a

b ab

b b

c c

0 150

B 100

3.2. Effect of the EDTA on the growth of B. napus

c c

Whole plant dry weight (g)

+ EDTA

Results showed that the addition of EDTA (3 g/kg) in 8 weeks old plants of B. napus cultivated in the presence of different sludge doses, did not modify significantly plant biomass production in this species during the treatment period (Fig. 2). Additionally, plant morphology did not show any modification due to the addition of EDTA in the soil as compared to those cultivated in the absence of this chelator. However, in plants subjected to 40 and 80 t/ha doses of sludge, we noticed that the addition of EDTA induced a slight increase of biomasses production at the final harvest (8 weeks after EDTA application).

50

b b a

0 150

a

a

a

C

e

d

100 + EDTA

140

50

Sludge: 80t/ha

Whole plant dry weight (g)

Sludge: 40t/ha Sludge : 20t/ha 105

b

e

c c b

b

b

0 150

Sludge : 0 t/ha

D

d 70

c

f

g

100

c 35

ab

b

+ EDTA

b

a a

a

0 8

10

c c

50

ab

ab

12

14

16

Different harvests period (weeks) Fig. 1. Variation of plant biomass production in Brassica napus cultivated in the presence of different sludge doses without addition of EDTA in the soil. Data are the means of six replicates. Values marked with the same letter are not significantly different at p = 0.05.

b

b b

0 8

10

12

14

16

Different harvests (weeks) Fig. 2. Effect of EDTA on growth of Brassica napus cultivated in the presence of different sludge doses. (A) Without sludge, (B) sludge: 20 t/ha, (C) sludge: 40 t/ha and (D) sludge: 80 t/ha. Means of six replicates and values marked with the same letter are not significantly different at p = 0.05.

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3.3. Heavy metals accumulation

250

3.3.2. Effect of EDTA on shoots Zn2+, Mn2+ and Pb2+ accumulation Zn2+, Pb2+ and Mn2+ shoot concentrations given in Fig. 4 are those of the final harvest (16 weeks). In presence of sludge, EDTA application increased Zn, Mn and Pb concentrations in the shoots. This effect was more pronounced at the highest dose of sludge 80 t/ ha (Fig. 4). The comparison between three metals shows that the

-EDTA +EDTA

Pb, µg/gDW

200

e d

150

d

c 100 50

b

b a

a

0 250

d

200

Zn, µg/gDW

3.3.1. Effect of sludge on shoot Zn2+, Mn2+ and Pb2+ accumulation In plant cultivated in unamended soil, shoot Zn, Mn and Pb concentrations were respectively on the average of 22.3, 17.5 and 27.76 lg/g DW. The sewage sludge application increased significantly plant Zn, Mn and Pb uptake (Fig. 3). The effect of sludge on the accumulation of metals on aboveground tissues increased with the increasing sludge doses and the period of treatment for the three metals. However, for both Pb and Mn, shoot concentrations were stabilized at respectively values of 117.6 and 112.3 lg/g DW after 12 weeks of treatments. Zn concentration in the shoot is positively correlated with the sludge doses and treatment duration, reaching 142.67 lg/g DW at the final harvest (16 weeks after sowing).

d

150

c b

100 50

e

c

a

a

0 250

150

0 t/ha

20 t/ha

40 t/ha

e

80 t/ha

200

Mn, µg/gDW

d

Pb, µg/gDW

100

50

150

c 100 50

12

14

a

a 0

20

40

80

Sludge amount, t/ha

150

Zn, µg/gDW

b

0

0

100

Fig. 4. Effects of EDTA on Pb, Zn and Mn shoot concentrations in Brassica napus plants cultivated during 8 weeks in soil amended with different sludge doses. Means of six replicates and values marked with the same letter are not significantly different at p = 0.05.

50

more important increases by EDTA concern Mn compared to Zn, and Pb.

4. Discussion

0 150

100

Mn µg/gDW

c

c

50

0

12

14

8

10

16

12

18

14

16

Treatment duration Fig. 3. Variation of Pb, Zn and Mn shoot concentrations in Brassica napus with different sludge dose in the absence of EDTA, means of six replicates and values marked with the same letter are not significantly different at p = 0.05.

With respect to the investigated heavy metals concentrations in sludge, ours data confirm the possibility of sludge use in agriculture. In fact, Cd2+, Pb2+, Zn2+ and Mn2+ sludge concentrations which are 2, 301, 515 and 234 (Table 1), respectively, does not exceeded the regulated limits allowed in agriculture amendment which are, 20, 800, and 300 ppm respectively for Cd2+, Pb2+, Zn2+ and Mn2+ (Wallace and Wallace, 1994; ADEME, 2001). The beneficial effects of sewage sludge addition on plant growth is evident in our experiment because we showed a significant increase of biomass production in B. napus cultivated in the presence of sludge as compared to control plants cultivated in unamended soil. This effect was accentuated with increasing sludge amounts (Fig. 1), suggesting that this amendment presents any toxic effect on plant growth, even at 80 t/ha. This result may be ascribed to the fertilizing effect of sewage sludge due to its high organic matter and macronutrients concentrations. The same result was found

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by Singh and Sinha (2005) and was attributed to the effect of sludge on soil fertility by the enhancement of the availability of some nutrients as N, P, K, Mg. . . and through the improvement of the water holding capacity of soil. Accordingly, beneficial effects of sludge on plant growth were also demonstrated by Roberts et al. (1988) and Pigozzo et al. (2000), respectively, in ray grass, fescue and maize. Cameron et al. (1997) and Abou Seeda (1997) stated that organic wastes such as sludge are valuable source of plant nutrients especially N, P and organic substrates, and could, therefore, positively affect the physical, the chemical and the biological properties of soil (Santos and Bettiol, 2003; Bettiol and Camargo, 2006; Singh and Agrawal, 2008). Similar results were observed by Zaman et al. (1999) Tawfik and Gomaa (2005) indicated that application of organic fertilizer greatly increased all growth parameters, grain yield and protein contents of the grain. A new concept for sewage sludge management involves its use as a fertilizer for crop production, including field and vegetable crops (Sermviriyakul, 2004). Sewage sludge contains considerable amounts of nutritive value especially due to high nitrogen content (Kalembasa et al., 2001; Tsadilas et al., 2005). Sewage sludge from dairy plant has a low C/N ratio (Fidecki, 2002). However, due to its origin (urban and industrial used water), sludge could contain high concentrations of heavy metals (Gupta and sinha, 2007; Ailincâi et al., 2008). Accumulation of these toxic elements in the upper layers of soils and in crop plants, due to the repetitive use of swage sludge as fertilizer, represent the principal limit of this practice (Nyamangara and Mzezewa, 1999). Toxic metals accumulation in aerial part of plants was observed in several species cultivated on soil amended with sludge. In fact, Rejeb and Bahri (1995), Singh and Sinha; (2005) detected a high concentrations of heavy metals, respectively, in Lactuca sativa and Brassica juncea cultivated in the presence of sewage sludge. More recently, Jamali et al. (2009) demonstrated the possibility of toxic heavy metals accumulation in wheat cultivated on soil dressed with untreated sewage sludge. Theses micro-pollutants are not biodegradable, so they must be extracted. In our case, although the relative low heavy metals sludge concentration, we made a great attempt to extract heavy metal. We tested the possibility of B. napus use to accumulate metals in harvestable parts from sludge enriched soils. Results showed that Zn, Mn and Pb shoot concentrations in plants presented a significant increase in the presence of sludge as compared to unamended soil (Fig. 3). Our results indicated that, although the positive correlation between sludge amounts and metals shoot accumulation, B. napus plants does not exhibited any toxic symptom like chlorosis or growth reduction. These data suggest that this species tolerate the analysed heavy metals. Furthermore, the plant ability to compartmentalize heavy metal in shoots suggests its use to remove theses pollutants from soil. Accordingly, Marchiol et al. (2004) demonstrated the possibility of the use of this species to absorb and accumulate metals from contaminated sites. Biomass of these plants could be used in industrial purposes (Veerle Grispen et al., 2006). On the other hand, in order to ameliorate the phytoextraction potential, the addition of synthetic metals chelators, as EDTA, is a reliable solution. This type of extraction is named assisted-phytoextraction (Zhou et al., 2007). Researchers demonstrated that this chelator enhances significantly metals availability on soil solution and so facilitates its absorption by the roots of plants and its translocation from the roots to the shoots (Wu et al., 2004; Meers et al., 2005, 2008). For EDTA assisted-phytoextraction, several others studies demonstrated the enhancement of heavy metals availability in soils and accumulation in harvestable organs of plants (Greˇman et al., 2001, 2003; Ulrich Schmidt, 2003; Nowack et al., 2006; Quartacci et al., 2007). However, the applied concentration of this chelator must be appropriate and not toxic for plant growth and microbial biomass. In fact some studies suggested that the

addition of this synthetic chelator may had a significantly adverse effect on plant growth (Lai and Chen, 2005; Quartacci et al., 2006). In our study, 3 g/kg soil of EDTA does not affect significantly B. napus growth in the absence and in presence of sludge in the soils. This data suggests that this dose of EDTA has not any deleterious effects on B. napus growth. However, this concentration of EDTA enhanced significantly shoots accumulations of Zn, Mn and Pb from sludge-amended soils, especially at 40 and 80 t/ha of sludge doses. We suggest that the total amounts of extracted metals will be more elevated in the presence of EDTA because this chelator enhanced shoots metals concentration without effects on plant growth. This result was consistent with others studies showing that the enhancement of heavy metals availability in soils by the addition of EDTA improves significantly their phytoextraction (Liphadzi and Kirkham, 2006; Van Engelen et al., 2007). In fact, Lai and Chen (2005) found that, in a soil artificially polluted with several heavy metals, EDTA application induced the accumulation of significant amounts of Cd2+ and Pb2+ in shoots of Dianthus chinensis. This result could be attributed to the effect of EDTA on the absorption and translocation of these metals on their complexed form metal–EDTA (Wu et al., 2004). More recently, it was reported that the accumulation of Cd in plant shoots correlated with the formation of the Cd–EDTA complex, and that Cd–EDTA was the major form of Cd absorbed and translocated by the plant (Van Engelen et al., 2007). The enhancement of metals accumulation and extraction by B. juncea plants in the presence of EDTA can reduce significantly the contamination of agricultural soils due to the application of sewage sludge as fertilizer. We suggest that used concentration of EDTA (3 ppm of dry soil) had not any negative effect on plant growth but enhanced significantly metal extraction.

5. Conclusion Owing to its fertilizing effects, sludge induced a significant improvement of B. napus growth. This effect was accompanied with metals (Zn2+, Pb2+ and Mn2+) accumulation in the shoots. Without effect on plant growth, the application of EDTA in sludge-amended soils enhanced significantly metals uptake and translocation in this species. So, as a whole, B. napus cultivation on sewage sludge-amended soils may be a promising solution to efficiently reduce toxic metal concentrations in soils and avoid the risk of groundwater and food heavy metal-induced contamination.

References Abou Seeda, M., 1997. Use of sewage sludge for sustainable agriculture and pollution preservation. 3. Treatment sewage sludge and its effect on chemical characteristics of sludge, soil and some nutrients uptake by radish, Spanish and lettuce plants. J. Agric. Sci. Mansoura Univ. 22 (10), 3423–3450. ADEME, 2001. Hygiénisation des bio-déchets: Validation du caractère hygiénisant de procédés de traitement. Déportes I. et Gay J. Rapport ADEME/OSER, p. 86. Ailincâi, C., Tsadilas, C.D., Sgouras, I.D., Bucur, D., Despina, A., Adriana, B., 2008. Influence of different organic resources on crop yield and soil and fertility in the Moldavian plateau. Cerceta˘ri Agronomice în Moldova 134, 23–34. Aldrich, M.V., Gardea-Torresdey, J.L., Peralta-Videa, J.R., Parsons, J.G., 2003. Uptake and reduction of Cr(VI) to Cr(III) by mesquite (Prospis spp.): chromate-plant interaction in hydroponics and solid media studied using XAS. Environ. Sci. Technol. 37, 1859–1864. Arthur, E.L., Rice, P.J., Rice, P.J., Anderson, T.A., Baladi, S.M., Henderson, K.L.D., Coats, J.R., 2005. Phytoremediation. Crit. Rev. Plant Sci. 24, 109–122. Bettiol, W., Camargo, O.A., 2006. Lodo de esgoto: impactos ambientais na agricultura. Jaguariuna: Embrapa Meio Ambiente, p. 349. Blaylock, M.J., Salt, D.E., Dushenkov, S., Zakharova, O., Gussman, C., Kapulnik, Y., Ensley, B.D., Raskin, I., 1997. Enhanced accumulation of Pb in Indian mustard by soil applied chelating agents. Environ. Sci. Technol. 31, 860–865. Brooks, R.R., 1994. Plants that hyperaccumulate heavy metals. In: Farago, M.E. (Ed.), Plants and the Chemical Elements: Biochemistry, Uptake, Tolerance and Toxicity. VCH, Weinheim, Germany, pp. 88–105.

H. Zaier et al. / Bioresource Technology 101 (2010) 3978–3983 Cameron, K.C., Di, H.J., McLoren, R.G., 1997. Is soil an appropriate dumping ground for our waste. Aust. J. Soil Res. 33, 995–1035. Cho, Y., Bolick, J.A., Butcher, D.J., 2009. Phytoremediation of lead with green onions (Allium fistulosum) and uptake of arsenic compounds by moonlight ferns (Pteris cretica cv Mayii). Microchem. J. 91, 6–8. Collins, R.N., Merrington, G., McLaughlin, M.J., Knudsen, C., 2002. Uptake of intact zinc-ethylenediaminetetraacetic acid from soil is dependent on plant species and complex concentration. Environ. Toxicol. Chem. 21, 1940–1945. Cooper, E.M., Sims, J.T., Cunnigham, S.D., Huang, J.W., Berti, W.R., 1999. Chelate assisted phytoextraction of lead from contamined soils. J. Environ. Qual. 28, 1709–1719. Cottenie, A.M.V., Kiekens, L., Camerlynck, R., Velgh, G., Dhaese, A., 1983. Essential and non essential trace elements in the system soil–water–plant. Laboratory Analytical Agrochemical State University, Ghent, Belgium. p. 75. Egner, H., Riehm, H., Domingo, W.R., 1960. Untersuchungen uber die chemische Bodenanalyse als Grundlage fur die Beurteilung des Nahrstoffzustandes der Boden, II. Chemische Extraktionsmethoden zur Phosphor-und Kalium bestimmung. Kungl. Lantbrukshogsko. Annal. 26, 45–46. Evangelou, M.W.H., Ebel, M., Schaeffer, A., 2007. Chelate assisted phytoextraction of heavy metals from soil: effect, mechanism, toxicity and fate of chelating agents. Chemosphere 68, 989–1003. Fidecki, M., 2002. Nutritive Value of Sewage Sludge from a Dairy Plant. Ph.D. Thesis. AR Lublin (in Polish). Gardea-Torresdey, J.L., Peralta-Videab, J.R., De La Rosaa, G., Parsons, J.G., 2005. Phytoremediation of heavy metals and study of the metal coordination by X-ray absorption spectroscopy. Coord. Chem. 249, 1797–1810. Greˇman, H., Velikonja-Bolta, Vodnik, D., Kos, B., Le tan, D., 2001. EDTA enhanced heavy metal phytoextraction: metal accumulation, leaching and toxicity. Plant Soil 235, 105–114. Greˇman, H., Vodnik, D., Velikonja-Bolta, S., Lestan, D., 2003. Ethylenediaminedisuccinate as a new chelate for environmentally safe enhanced lead phytoextraction. J. Environ. Qual. 32, 500–506. Gupta, A.K., Sinha, S., 2006. Chemical fractionation and heavy metal accumulation in the plant of Sesamum indicum (L.) var. T55 grown on soil amended with tannery sludge: selection of single extractants. Chemosphere 64, 161–173. Gupta, A.K., Sinha, S., 2007. Phytoextraction capacity of the Chenopodium album L. Grown on soil amended with tannery sludge. Biores. Technol. 98, 442–446. Hernandez, T., Moreno, J.I., Costa, F., 1991. Influence of sewage sludge application on crop yields and heavy metal availability. Soil Sci. Plant Nutr. 37, 201–210. Hernandez-Allica, J., Barrutia, O., Becerril, J.M., Garbisu, C., 2003. EDTA reduces the physiological damage of lead on cardoon plants grown hydroponically. J. Phys. IV 107, 613–616. Jamali, M.K., Kazi, T.G., Arain, M.B., Afridi, H.I., Jalbani, N., Kandhro, G.A., Shah, A., Baig, J.A., 2009. Heavy metal accumulation in different varieties of wheat (Triticum aestivum L.) grown in soil amended with domestic sewage sludge. J. Hazard Mater. 164, 1386–1391. Kalembasa, S., Kalembasa, D., Kania, R., 2001. Zesz. Probl. Post. Nauk Roln. vol. 475, p. 279. Kalra, Y.P., Maynard, D.G., 1991. Methods for Forest Soil and Plant Analysis. Information Report NOR-X-319. Forestry Canada, Northwest Region, Northern Forestry Center, p. 116. Lai, H., Chen, Z., 2005. The EDTA effect on the phytoextraction of single and combined metals contaminated soils using rainbow pink (Dianthus chinensis). Chemosphere 60, 1062–1071. Liphadzi, M.S., Kirkham, M.B., 2006. Availability and plant uptake of heavy metals in EDTA-assisted phytoremediation of soil and composted biosolids. S. Afr. J. Bot. 72, 391–397. Lombi, E., Zhao, F.J., Dunham, S.J., McGrath, S.P., 2001. Phytoremediation of heavy metal-contaminated soils: natural hyperaccumulation versus chemically enhanced phytoextraction. J. Environ. Qual. 30, 1919–1926. Lotte, V.N., Jan, M., Koen, O., Kris, V., 2007. Phytoextraction of metals from soils: how far from practice? Environ. Pollut. 150, 34–40. Luo, C.L., Shen, Z.G., Li, X.D., 2008. Plant uptake and leaching of metals during the hot EDDS-enhanced phytoextraction process. Int. J. Phytorem. 9, 181–196. Marchiol, L., Assolari, S., Sacco, P., Zerbi, G., 2004. Phytoextraction of heavy metals by canola (Brassica napus) and radish (Raphanus sativus) grown on multicontaminated soil. Environ. Pollut. 132, 21–27. McGrath, S.P., Cunliffe, C.H., 1985. A simplified method for the extraction of the metals Fe, Zn, Cu, Ni, Pb, Cr, Co and Mn from soils and sewage sludges. J. Sci. Food Agric. 36, 794–798. McGrath, S.P., Zhao, F.J., Lombi, E., 2002. Phytoremediation of metals, metalloids, and radionuclides. Adv. Agron. 75, 1–56. Meers, E., Hopgood, M., Lesge, E., Vervake, P., Tack, F.M.G., Verloo, M.G., 2005. Comparison of EDTA and EDDS as potential soil amendments for enhaced phytoextraction of heavy metal. Chemosphere 58, 1011–1022. Meers, E., Tack, F.M.G., Verloo, M.G., 2008. Degradability of ethylenediaminedisuccinic acid (EDDS) in metal contaminated soils: implications for its use soil remediation. Chemosphere 70, 358–363. Nowack, B., 2002. Environmental chemistry of aminopolycarboxylate chelating agents. Environ. Sci. Technol. 36, 4009–4016. Nowack, B., Schulin, R., Robinson, B.H., 2006. Critical assessment of chelantenhanced metal phytoextraction. Environ. Sci. Technol. 40, 5225–5232. Nyamangara, J., Mzezewa, J., 1999. The effect of long-term sewage sludge application on Zn, Cu, Ni and Pb levels in a clay loam soil under pasture grass in Zimbabwe. Agric. Ecosyst. Environ. 73, 199–204.

3983

Peer, W.A., Baxter, I.R., Richards, E.L., Freeman, J.L., Murphy, A.S., 2005. Phytoremediation and hyperaccumulator plants. In: Tamas, M., Martinoia, E. (Eds.), Molecular Biology of Metal Homeostasis and Detoxification. Topics in Current Genetics. Springer, Berlin, vol. 14, pp. 299–340. Petra, K., Juan, B., Pilar Bernal, M., Flavia, N., Charlotte, P., Stefan, S., Rafael, C., Carmela, M., 2009. Trace element behaviour at the root–soil interface. Implications in phytoremediation. Environ. Exp. Bot. 67, 243–259. Pigozzo, A.T.J., Gobbi, M.A., Lenzi, E., Luchese, E.B., 2000. Effects of the application of sewage sludge and petrochemical residue in maize culture as source of micronutrients on soils of Parana state. Braz. Arch. Biol. Technol. 43, 143–149. Quartacci, M.F., Argilla, A., Baker, A.J.M., Navari-Izzo, F., 2006. Phytoextraction of metals from a multiply contaminated soil by Indian mustard. Chemosphere 63, 918–925. Quartacci, M.F., Irtelli, B., Baker, A.J.M., Navari-Izzo, F., 2007. The use of NTA and EDDS for enhanced phytoextraction from a multiply contaminated soil by Brassica carinata. Chemosphere 68, 1920–1928. Rejeb, S., Bahri, A., 1995. Incidence de l’apport de boues résiduaires urbaines sur la composition minérale et la productivité de quelques espèces cultivées en Tunisie. Les Cahiers du CRGR 24, 13–31. Renoux, A.Y., Rocheleau, S., Sarrazin, M., Sunahara, G.I., Jean-Francois, B., 2007. Assessment of a sewage sludge treatment on cadmium, copper and zinc bioavailability in barley, ryegrass and earthworms. Environ. Pollut. 145, 41–50. Roberts, J.A., Daniels, W.L., Bell, J.C., Martens, D.C., 1988. Tall fescue production and nutrient status on southwest Virginia mine soils. J. Environ. Qual. 17, 55–61. Römkens, P., Bouwman, L., Japenga, J., Draaisma, C., 2002. Potentials and drawbacks of chelate-enhanced phytoremediation of soils. Environ. Pollut. 116, 109–121. Salt, D.E., Smith, R.D., Raskin, I., 1998. Phytoremediation. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49, 643–668. Santos, I., Bettiol, W., 2003. Effect of sewage sludge on the rot and seedling damping-off of bean plants caused by Sclerotium rolfsii. Crop Prot. 22, 1093– 1097. Sermviriyakul, K., 2004. The Study of the Substitution of Sewage Sludge for Chemical Fertilizer in Soybean Production. Master Thesis, National Institute of Development Administration, Bangkok, Thailand. Shen, Z.G., Li, X.D., Wang, C.C., Chen, H.M., Chua, H., 2002. Lead phytoextraction from contaminated soil with high-biomass plant species. J. Environ. Qual. 31, 1893–1900. Singh, R.P., Agrawal, M., 2007. Effects of sewage sludge amendment on heavy metal accumulation and consequent responses of Beta vulgaris. Chemosphere 67, 2229–2240. Singh, R.P., Agrawal, M., 2008. Potential benefits and risks of land application of sewage sludge. Waste Manage. 28, 347–358. Singh, S., Sinha, S., 2004a. Morphoanatomical response of two cultivars of Brassica juncea (L.) Czern grown on tannery waste amended soil. Bull. Environ. Contamin. Toxicol. 72, 1017–1024. Singh, S., Sinha, S., 2004b. Scanning electron microscopic studies and growth response of the plants of Helianthus annuus L. Grown on tannery sludge amended soil. Environ. Int. 30, 389–395. Singh, S., Sinha, S., 2005. Accumulation of metals and its effects in Brassica juncea (L.) Czern. (cv. Rohini) grown on various amendments of tannery waste. Ecotox. Environ. Safety 62, 118–127. Tandy, S., Schulin, R., Nowack, B., 2006. The influence of EDDS on the uptake of heavy metals in hydroponically grown sunflowers. Chemosphere 62, 1454–1463. Tawfik, M.M., Gomaa, A.M., 2005. Effect of organic and bio fertilizers on the growth and yield of wheat plants. Egypt J. Agric. Res. 2, 711–725. Tsadilas, C.D., Matsi, T., Barbayiannis, N., Dimoyiannis, D., 2005. Influence of Sewage Sludge Application on Soil Properties and the Distribution and Availability of Heavy Metal Fraction, Soil Science Laboratory, Aristotelian University, Thessaloniki, Greece, pp. 2603–2618. Schmidt, Ulrich., 2003. Enhancing phytoextraction: the effect of chemical soil manipulation on mobility, plant accumulation, and leaching of heavy metals. J. Environ. Qual. 32, 1939–1954. Van Engelen, D.L., Sharpe-Pedler, R.C., Moorhead, K.K., 2007. Effect of chelating agents and solubility of cadmium complexes on uptake from soil by Brassica juncea. Chemosphere 68, 401–408. Vassil, A.D., Kapulnik, Y., Raskin, I., Salt, D.E., 1998. The role of EDTA in lead transport and accumulation by Indian mustard. Plant Physiol. 117, 447–453. Veerle Grispen, M.J., Hans Nelissen, J.M., Jos Verkleij, A.C., 2006. Phytoextraction with Brassica napus L. A tool for sustainable management of heavy metal contaminated soils. Environ. Pollut. 144, 77–83. Wallace, A., Wallace, G.A., 1994. A possible flow in EPA’s 1993 new sludge rule due to heavy metal interactions. Commun. Soil Sci. Plant Anal. 25, 129–135. Wenger, K., Gupta, S.K., Furrer, G., Schulin, R., 2003. The role of nitrilotriacetate in copper uptake by tobacco. J. Environ. Qual. 32, 1669–1676. Wu, L.H., Luo, Y.M., Xing, X.R., Christie, P., 2004. EDTA-enhanced phytoremediation of heavy metal contaminated soil with Indian mustard and associated potential leaching risk. Agric. Ecosyst. Environ. 102, 307–318. Zaman, M., Di, H.J., Cameron, K.C., Frampton, C.M., 1999. Gross nitrogen mineralization and nitrification rates and their relationships to enzyme activities and the soil microbial biomass in soil treated with dairy shed effluent and ammonium fertilizer at different water potential. Biol. Fertil. Tilt Soils 29, 178–186. Zhou, D.M., Chen, H.F., Cang, L., Yu-Jun, W., 2007. Ryegrass uptake of soil Cu/Zn induced by EDTA/EDDS together with a vertical direct-current electrical field. Chemosphere 67, 1671–1676.