You are on page 1of 10

J Soils Sediments (2012) 12:900–908

DOI 10.1007/s11368-012-0520-2

SOILS, SEC 3 I REMEDIATION AND MANAGEMENT OF CONTAMINATED OR DEGRADED LANDS I RESEARCH ARTICLE

Bioleaching of heavy metals from sewage sludge
by Acidithiobacillus thiooxidans—a comparative study
Ye-Ming Wen & Qing-Ping Wang & Caixian Tang &
Zu-Liang Chen

Received: 19 December 2011 / Accepted: 1 April 2012 / Published online: 27 April 2012
# Springer-Verlag 2012

Abstract
Purpose To understand the bioleaching of metals from
sludge by Acidithiobacillus thiooxidans, the aims of this
study were to evaluate the experimental conditions affecting
the efficiency of removal of the metals, including solids
concentration, initial pH, sulfur concentration and inoculum
level were examined, and following the bioleaching mechanism was proposed.
Materials and methods A. thiooxidans were isolated from
collected sludge samples containing bacteria from Fuzhou
Jingshan sewage treatment plant, and identification of bacteria
by sequencing the 16 s rDNA gene sequences. Conditions
affecting the bioleaching and application were conducted by
batch experiments. The analysis of Cr, Cu, Pb, and Zn was
carried out using an atomic absorption spectrophotometer, and
the pH and oxidation–reduction potential (ORP) were measured using a pH meter and an ORP meter.
Results and discussion The results show that a high metal
leaching efficiency was achieved at low solid concentrations
due to decreases in buffering capacity. In addition, the best
conditions of the bioleaching included 2 % (w/v) solid concentration, 5.0 gL−1 sulfur concentration, and 10 % (v/v)
inoculum concentration, where the removal efficiencies of
Responsible editor: Bernd Markert
Y.-M. Wen : Q.-P. Wang : Z.-L. Chen (*)
School of Environmental Science and Engineering,
Fujian Normal University,
Fuzhou 350007 Fujian Province, People’s Republic of China
e-mail: zlchen@fjnu.edu.cn
C. Tang
Department of Agricultural Sciences, La Trobe University,
Bundoora,
Melbourne, VIC 3086, Australia

Cr, Cu, Pb, and Zn in sewage sludge was 43.6 %, 96.2 %,
41.6 %, and 96.5 %, respectively.
Conclusions We found that the bioleaching of Zn was governed by direct and indirect mechanisms, while the bioleaching of Cu, Pb, and Cr was mainly dominated by the
bioleaching indirect mechanism. After processing with the
proposed techniques, the heavy metals in the sewage sludge
did meet the requirement of the national standards.
Keywords Acidithiobacillus thiooxidans . Bioleaching .
Heavy metal . Sewage sludge

1 Introduction
In recent years, the quantity of the total sludge generated all
over the world has increased dramatically, the disposal of
which is one of the serious environmental concern (Babel
and Dacera 2006; Pathak et al. 2009; Lombardi et al. 2001).
Since the origin of the sewage and its treatment from the
sewage treatment plant varies, and sewage sludge contains
high concentrations of toxic metals (Lombardi and Garcia
2002), the disposal of untreated sludge to land shows a
potential hazard to human health and to the environment
(Tsai et al. 2003). For these reasons, it is necessary to remove
the heavy metals from the sludge before being used on land.
Various chemicals such as chelating agents such as ethylene
di-amine tetraacetic acid and nitrilotriacetic acid (Lo and Chen
1990) have been reported for extraction of metals from sludge.
However, the practical application of the chemical processes
is still limited due to the requirement of large amount of
chemicals, the high operating cost, the operational difficulties,
and the secondary pollution problems associated with them
(Pathak et al. 2009; Kumar and Nagendran 2008). As such,

0. The oxidation and acid production by A. the characteristics of the metal.66 Zn Cu Cr Pb 1. energy source. thiooxidans are the primary impetuses of solubilization of heavy metals (Zhang et al. the different forms of heavy metal in the sludge vary according to the type of sludge.J Soils Sediments (2012) 12:900–908 the microbial method is one of the options that can be used to remove heavy metals in the sludge due to its low cost and low energy requirement (Hsu and Harrison 1995). The influence of initial pH of the system on bioleaching of heavy metals from metalcontaminated soils has also been reported. Molecular identification of the isolated culture was done by sequencing the 16 s rDNA gene sequences.1 Sewage sludge source and its characterization The sludge sample containing bacteria was collected from Fuzhou Jingshan sewage treatment plant. 0. 2009). has been reported to be an efficient and economical method for removal of heavy metals from the sludge (Babel and Dacera 2006. where a decrease in pH. where the reports concerned only one condition impacting on the bioleaching such as the pH. thiooxidans. In bioleaching process. After three generations of enrichment. and the reduction of metal were associated with increases in the concentration and the buffering capacity of sludge solids (Chen and Lin 2004a.430 285 204 . solids concentration. The initial pH of the Waksman liquid medium was 4.9 7. leading to cause sludge acidification and to make heavy metal solubilization (Babel and Dacera 2006. thiooxidans have recently been reported.2 Isolation and identification of indigenous sulfur-oxidizing bacteria For enrichment of sulfur-oxidizing bacteria. Pathak et al. 2008). Bioleaching. an increase in oxidation–reduction potential (ORP). which is based on the ability of microorganisms to transform metals from the solid waste into soluble and extractable forms. The measured characteristics of the sludge sample were listed in Table 1. where A.2 g L−1 (NH4)2SO4. the isolate was inoculated on the Waksman solid medium. Ltd. and following. sulfur concentration.0 gL−1 K2HPO4. The effects of sludge solids concentration on the bioleaching process in a batch bioreactor have been reported. this study systematically investigated the bioleaching of Cr. A number of studies on bioleaching of heavy metals using A. The sludge sample was transported back to the laboratory and stored at 4°C prior to use. 10 mL of the acidified sludge was transferred to the second flask containing 100 mL of the Waksman liquid medium under the same conditions. 3. thiooxidans. and the pure culture for further use was obtained after three successive subcultures (Kumar and Nagendran 2007).5 gL−1 MgSO4·7H2O.6 95. which highlights that pH plays an important role in metal solubilization and that microorganisms can utilize elemental sulfur as an energy source (Kumar and Nagendran 2007). Cr. Pathak et al. Pb. and Zn in sewage sludge using A. and leaching microorganisms (Babel and Dacera 2006. 0. thiooxidans was demonstrated under the optimal conditions. 2 Materials and methods 2.9 10. thiooxidans oxides sulfur to sulfuric acid to obtain energy from the oxidation process. the Waksman liquid medium of the following composition was used: 0. 10 gL−1 S0 as an energy source. For these reasons. 2009). the pH of the medium was monitored. 2. Acidithiobacillus thiooxidans can be used in leaching of metals from sewage sludge. 2009). the bioleaching mechanism was proposed. Pb.6 40. and inoculum level were examined. However. including solids concentration. another part was used for bioleaching experiments.25 gL−1 CaCl2·2H2O. However. 901 The feasibility of the removal of metal ions such as Cu. Test sludge (10 mL with solids concentration of 4 % (w/v)) was taken in a 250-mL flask containing 100 mL of the Waksman liquid medium.0. The subsample was sterilized by autoclaving (30 min at 120°C) and was characterized. it is therefore necessary to systemically investigate various conditions affecting the bioleaching of heavy metals from sludge. this is limited to understanding the mechanisms of bioleaching because of its lacking evidence.732 2. Table 1 The selected characteristics of sewage sludge Characteristics Solids (%) Organic matter (%) Total P (g kg−1 dry sludge) Total N (g kg−1 dry sludge) pH Heavy metals (mg kg−1 dry sludge) 17. Cu. When the pH reached 2. The experimental conditions affecting the efficiency of removal of the metals.. SKY-2102) and shaken at 150 rpm at 30°C. To understand the bioleaching process. The flasks were mounted on a shaking incubator (Shanghai Sukun CO. For example. Pathak et al. and the method employed for the treatment of the sewage. and Zn from sewage sludge using A. the effect of sulfur on bioleaching of heavy metals from mine tailings bacteria has been recently reported. the results have indicated that 2 % sulfur substrate concentration is optimal for bacterial activity and metal solubilization (Liu et al. initial pH. b). 2009). The sample was divided into two parts. One part was used for isolating A.

In contrast with the trend of change in pH.30 to 1. which was based on homology of its 16 S rDNA gene sequences from the National Center for Biotechnology Information databank using BLASTN (Lin et al. ORP increased rapidly from 300 to 445 mV in the first 4 days of bioleaching. A rapid drop in pH was observed in the first 6 days. Cu. 6 %. microscopic examination of the culture revealed the presence of gram negative. the ORP of the control was kept around 295 mV during the same bioleaching period. Ltd. 4.. leading to decreased concentration of sulfur followed by increases in ORP (Kumar and Nagendran 2008). the pH decreased gradually to 4. The factors affecting the metal leachability using A. the pH of the sludge medium decreased with increasing bioleaching time as shown in Fig. A control run without inoculum of bacteria was also carried out in order to compare the results. Pb. 2009. 5.. then increased to 5. 10. 8 %. respectively. which was agitated at 30°C and 150 rpm on a gyratory shaker for 24 h to raise its temperature and initial ORP. A 10 % volume of active bacterium suspension was added to the sludge slurry with an initial cell concentration of 108~109 cells per milliliter and followed by the addition of 0. However. the water loss due to evaporation was replenished with distilled water daily. Varian. thiooxidans. rod shaped. The whole set-up was weighed and incubated as described above. thiooxidans accelerated the sludge acidification resulted from oxidizing sulfur to sulfuric acid and an increase in the ORP due to a stimulated microbial activity.75 g elemental sulfur. Consequently.07 due to the microbial oxidation of sulfur and the production of sulfuric acid based on their bioleaching mechanisms (Pathak et al. a sludge slurry (solids concentration. and its alkaline range observed in the control can be attributed to buffering action and release of carbonate substance from the sewage sludge with basic nature (Kumar and Nagendran 2008). as shown in Fig.45-μm membrane filter. 2a.000 rpm for 10 min. After 12 days of bioleaching. the final pH was stabilized at about 4.2 Changes in pH and ORP during bioleaching The changes in pH and ORP during the bioleaching process are shown in Fig. 15 %. As shown in Fig. Suzuki 2001).3 Experimental conditions affecting the bioleaching and application Laboratory scale bioleaching experiments were carried out in 250-mL Erlenmeyer flasks containing 150 mL of the sludge samples. sulfur concentration was 5 gL−1. five levels of sulfur (2. A slight increase in pH observed in the control indicates that chemical oxidation of sulfur did not occur. Thiooxidans in the sludge was able to alter oxided sulfur to sulfuric acid during their growth. and the supernatant was filtered through a 0. 4 %. increased slightly to 505 mV in the following 10 days and was then maintained until the end of bioleaching. ORP-421). and 6) which were achieved by adjustment using 2 N H2SO4. Similar results were reported on the study of the pH profile during bioleaching of metals resulted from anaerobical digestion by sulfur-oxidizing bacteria (Villar and Garcia 2006). All experiments were performed in replicate and were incubated in a gyratory incubator at 30°C and 150 rpm.4 Analytical methods The pH and ORP were measured using a pH meter (Shanghai Precision and Scientific Instrument Co. 3. thiooxidans A. 150 rpm for 24 h. The samples were centrifuged at 7. The data showed that the isolated J Soils Sediments (2012) 12:900–908 microorganism was closely related to acidophilic sulfuroxidizing bacteria A. These results show that the inoculation of A. 2b. 5 %. thiooxidans includes five solids concentrations (2 %. These results provide the evidence for the indirect bioleaching mechanism by A. where the sulfur in the sludge is oxidized . Ltd. five initial pH (2.62 in the first 6 days. During the bioleaching process. To test the removal efficiency of metals under the optimal conditions. 10 %. when sludge concentration was 2 %. 2. thiooxidans. where the pH was reduced from 5. and motile bacteria.902 2. 3. However. thiooxidans isolated from the sewage sludge grew well on a medium containing elemental sulfur as the energy source..42 in the following 2 days. The high values of ORP together with the low pH are responsible for the solubilization of metals (Lombardi and Garcia 2002). 3 Results and discussion 3. and 20 g L−1) and five inoculum concentrations of 108~109 cells/mL (2 %. The activated A. USA). which provided a suitable condition for the growth of A. and 10 % (w/v)).1 Identification of A. Samples (15 mL) were collected at 2-day intervals from the flasks for analyses of heavy metals. thiooxidans with 99 % similarity by the classification. 5. 2 % (w/v)) of 135 mL contained in a 250-mL flask was agitated at 30°C. thiooxidans. 2.98. the pH of the control was higher than those of the test treatments. The analysis of Cr. and Zn was carried out using an atomic absorption spectrophotometer (AA240. 2010). 15. This is because the elemental sulfur was oxidized to soluble sulfate. This also indicates that the sulfur can be used as the energy source for the growth of A. and inoculum concentration was 10 %.. PHS-3 C) and an ORP meter (Shanghai Kang-Yi Instruments Co. and 20 %). in the control medium. 1.

inoculum concentration.3 % Cu. This is due to an increased precipitation of PbSO4 at a high solids concentration (Ryu et al.J Soils Sediments (2012) 12:900–908 903 Fig. 1995). the elemental sulfur in the sludge is oxidized by A. 30°C. and Cr from the sludge during the bioleaching process. the bioleaching efficiency clearly 6 a b 500 5 450 ORP (mv) 4 pH Fig. which reduces the pH of the sludge medium. thiooxidans MS þ 2O2 ! MSO4 3. agitation speed. Major factors are investigated in the following sections. thiooxidans with a high bioleaching efficiency of metals from the sludge. 3. Cu. Pb. Generally. thiooxidans into soluble metal sulfates according to the following reaction: A. Cu. the removal efficiency of Pb was substantially lower at high solids concentrations even with longer leaching times compared to other treatments. thiooxidans into sulfuric acid.3.t ORP control 350 2 300 1 0 2 4 6 Time (d) 8 10 12 250 0 2 4 6 Time (d) 8 10 12 . 81.3 Conditions affecting the bioleaching In bioleaching of metals from sludge using A. into sulfuric acid and leads to acidification of the sludge medium (Jain and Tyagi 1993. 75. the highest removal efficiency achieved was at 2 % solids. 47. thiooxidans S0 þ H2 O þ 1:5O2 ! H2 SO4 ð2Þ H2 SO4 þ materials  M ! materials  2H þ MSO4 ð3Þ where M is a bivalent metal. b. thiooxidans. 1 The colony morphology and individuals of A. 10 %. sulfur concentration. 2009). thiooxidans depends on physical.9 % Zn. To demonstrate the feasibility of using A. and 64. unlike other reports (Chen and Lin 2004a. thereby enhancing the solubilization of the metal: A. However.1 Effort of sludge solids concentration on bioleaching Figure 3 presents how sludge solids concentration affects the solubilization of Zn. In direct bioleaching. 150 rpm) pH A.t pH control 3 400 ORP A. chemical. 2 Dynamics of pH and oxidation–reduction potential (sludge concentration. For each metal.5 % Pb. Pb. both direct and indirect mechanisms have been proposed (Pathak et al. temperature. and biological factors in the system. Du et al. Kumar and Nagendran 2007). 2 %. and Cr decreased with increasing solid sludge concentration. the bioleaching efficiency of Zn. it is necessary to systematically understand the experimental conditions that affect the bioleaching process. For example. On the viewpoint of direct and indirect mechanisms. 5 gL−1. metal sulfides are directly oxidized by the A. Longer bioleaching times were required to produce a sufficient acidification of the leaching solution at high sludge solid concentrations. with 10 gL−1 sulfur concentration and 10 % inoculum concentration.7 % Cr in the sludge were removed in the first 6 days due to a faster reduction of pH at ð1Þ In indirect bacterial leaching. 2003).

2002). respectively. this current study shows that A. the removal efficiency varied only from 86. and 49. 4a is in agreement with the study on bioleaching of heavy metals from contaminated soil using A. In addition.904 100 100 a b 80 80 Removal of Cu (%) Removal of Zn (%) Fig. 3. the removal efficiency of Cu ranged from 83. inoculum concentration.1 %. and Pb was increased to 93. 1991). The removal efficiency for Zn. Cr.3 % at pH of 5 after bioleaching for 12 days. Another reason is that microorganisms are able to survive at higher solid contents of the sludge (Henry et al. and resulted in reduce of bioleaching solubilization. In the Zn approach.1 % at an initial pH of 2 to 40. b Cu. the removal of metals increased significantly with time and then plateaued at the values lower than 2 % solids concentration. the higher removal efficiency of Zn and Cu was observed at different initial pH. 91. 2002). c Pb. Low solubilization of Cu at initial pH 4 and 5 was reported while carrying out bioleaching of heavy metals from aquatic sediments employing Acidithiobacillus ferroxidans (Chartier and Couillard 1997).1 % after bioleaching for 12 days. Nevertheless. 4a. by day 12 using 2 % solids concentration. 64. temperature.2 %.2 Effort of initial pH on bioleaching Figure 4 shows the removal efficiency of the heavy metals at various initial pH values at 2 % sludge concentration.3 %.5 % Cr were removed at solids concentration of 10 %. and d Cr during the bioleaching process as affected by sludge solids concentrations (sulfur concentration. This is because high solids concentrations result in high buffering capacity. Similarly. However.4 % to 86. initial pH. 10 %. 2. 2007). leading to an increased time to attain a low pH (Xiang et al.2 % to 88. while the lowest value found at pH 2. Liu et al.66. agitation speed.7 % after bioleaching for 12 days.62×10−8) (Lo and Chen 1990). In comparison. 10 g 8 10 12 0 0 2 4 6 8 10 12 Time (d) L−1 a sulfur concentration and 10 % inoculum concentration. b. indicating the initial pH values did not significantly impact on the bioleaching of metals. The removal efficiency of Pb varied from 36.2 % Zn.1 %. the high range of solubilization of Zn as shown in Fig. 90. d). It is this reason that . 7.2 % Cu. the increase in concentration of the leached metals also results in toxicity that may inhibit the growth of the microorganisms (Cho et al. In contrast. Cu.3. 53. 2003). thiooxidans can be used effectively for bioleaching of heavy metals with the efficiency in the order of Zn>Cu>Cr>Pb. 2000.2 % Pb. 4c. However. 150 rpm) J Soils Sediments (2012) 12:900–908 60 40 2% 4% 6% 8% 10% 20 0 0 2 4 6 8 10 60 40 2% 4% 6% 8% 10% 20 0 12 0 2 4 Time (d) 6 8 10 12 Time (d) 60 c 2% 4% 6% 8% 10% 30 Removal of Cr (%) Removal of Pb (%) 60 40 20 10 0 d 70 50 2% 4% 6% 8% 10% 50 40 30 20 10 0 2 4 6 Time (d) this solid concentration (Ryu et al. 10 gL−1. the solubilization pattern of Pb and Cr was highly dependent on initial pH values (Fig. but there was slightly difference. and 18. 3 Solubilization of a Zn. thiooxidans (Kumar and Nagendran 2007). leading to complete cessation of metal bioleaching at solids concentration of 10 % (Cho et al. As shown in Fig. although the highest removal efficiency was recorded at an initial pH of 6. where variations in the initial pH of the system did not influence the bioleaching of Zn and Cu. 30°C. The overall low Pb removal with solution pH was mainly due to the precipitation of PbSO4 with a very low solubility (Ksp 01.

3 Effort of sulfur concentration on bioleaching As shown in Eq.5 to 15 gL−1. Figure 5 shows the effect of sulfur concentration on the removal efficiency 8 10 12 0 0 2 4 6 8 10 12 Time (d) of heavy metals when the sludge concentration of 2 %. the metals in sludge become soluble during bioleaching. inoculum concentration. while Cr in the range of 18. extremely acidic conditions tend to accelerate the solubilization of Cr (Fang and Zhou 2007). Since Cr solubilization is pH dependent. b). The similarly low solubilization of Pb during bioleaching of heavy metals from sludge employing sulfur-oxidizing bacteria was also observed (Couillard and Mercier 1993).5 and 15 gL−1 significantly increased the removal efficiency of metals after bioleaching for 2 days. b Cu. resulting in acidification of the sludge medium. and the inoculum concentration of 10 % were used. Chen and Lin 2001). In addition. 2 %. 3.3. Consequently.4 % at pH 2 to 64. Increasing sulfur concentration from 0. the batch study shows a major portion of the sulfur used in bioleaching was not completely oxidized (Chen and Lin 2004a. This is because Cu is mainly associated with sulfides/organic matter which can be directly oxidized by A. exchangeable. 30°C. thiobacilli.J Soils Sediments (2012) 12:900–908 100 100 a 60 Removal of Cu (%) 80 Removal of Zn (%) Fig. 3. It can be seen from Fig. Therefore. Cr and Pb data are identical where obtained at sulfur concentration of 0.1–99.5 was recommended for a sufficient removal of all the types of heavy metals at various sludge solids concentrations. leading to enhanced solubilization of metals (Chen and Lin 2004a. and Fe/Mn oxide fractions) (Chen and Lin 2000).5 % at pH 4 after 12 days of bioleaching. 10 gL−1. 150 rpm) 905 2 3 4 5 6 40 60 2 3 4 5 6 40 20 20 0 b 80 0 2 4 6 8 10 0 12 0 2 4 Time (d) 6 8 10 12 Time (d) 70 c 40 30 2 3 4 5 6 20 10 0 d 60 Removal of Cr (%) Removal of Pb (%) 50 50 40 2 3 4 5 6 30 20 10 0 2 4 6 Time (d) the efficiency of Pb solubilization was generally lower than other metals in the bioleaching process. The removal efficiency of Cu and Zn is often high from sludge (Chen and Lin 2004a.e. respectively. the determination of sulfur concentration is of great significance in developing an efficient bioleaching process. This may be due to the high pH buffer capacity or the release of alkaline from sewage sludge during the initial bioleaching (Liu et al. carbonate. thiooxidans at neutral pH without any initial pH adjustment (Ryu et al. the results suggest that bioleaching of metals is easily affected by pH.8 % and 84. while the bioleaching of Zn is found in more available and mobile fractions (i. The increase in the bioleaching efficiency of metal after 2 days of bioleaching is due to a fast acidification rate of sewage sludge. 4 Effect of initial pH on solubilization of a Zn.5– 99. the removal efficiency of Cr ranged from 59.6 %. The higher removal efficiency of Zn and Cu was achieved in the range of 90. 2003). A pH of 1–1. the low efficiency of Cr can be attributed to its low solubilization at extremely . 2008).4–66. However.5 % after bioleaching for 12 days. temperature. However. b).8 % and Pb in the range of 9. agitation speed. b). This finding is also supported by the study on the microbial leaching processes using A. c Pb. The bioleaching efficiency of Pb is generally lower than other metals because of the formation of insoluble PbSO4 as discussed above (Tyagi 1992..8– 43. and d Cr during the bioleaching process (sludge concentration. sulfur concentration. 10 %. the elemental sulfur is oxidized to form sulfuric acid. In addition. 5 that the removal efficiency of metals from sewage sludge was in the decreasing order of Zn>Cu>Cr>Pb.

. temperature.5 g/L 2 g/L 5 g/L 10 g/L 15 g/L 30 Removal of Cr (%) Removal of Pb (%) 6 Time (d) 20 0.5 g/L 2 g/L 5 g/L 10 g/L 15 g/L 40 20 0 12 0 2 4 Time (d) 50 70 c 8 10 12 8 10 12 d 60 40 0. 2005). and 66. agitation speed. Zn and Cu are easily soluble in sludge during the bioleaching even at a low initial inoculum concentration. A higher concentration of sulfur enables higher solubilization of metals via short-term bioleaching but is not recommended because un-oxidized sulfur remains in the sludge..8) after 12 days (see Fig.4 % after 12 days of bioleaching. inoculum concentration.2–44. Figure 6 shows the removal efficiency of heavy metals at various initial inoculum concentrations with the sludge concentration of 2 % and the sulfur concentration of 5 gL−1. 7. 2009). 95. The removal efficiency of Zn and Cu ranged from 92.4 Removal of heavy metals from the sewage sludge Bioleaching of Zn. 150 rpm) J Soils Sediments (2012) 12:900–908 60 0. c Pb.66.906 100 100 a 80 Removal of Cu (%) Removal of Zn (%) Fig. and also oxidize the elemental sulfur to form sulfuric acid. 99.8 % Zn. 8 10 12 0 0 2 4 6 Time (d) where the inoculum addition had been greater than 2. The removal efficiency of Zn. 2 %.0 %. 2003).5 g/L 2 g/L 5 g/L 10 g/L 15 g/L 40 20 0 0 2 4 6 8 10 b 80 60 0. Furthermore. 5 Solubilization of a Zn. and it was concluded that that sulfur was completely oxidized to sulfuric acid. 99. and 10 % inoculum concentration. leading to increases in bioleaching of Pb and Cr (Pathak et al. In contrast. 3. Hence. 5. and 43.7 % and 4.3. including 2 % solid concentration.0 %. and the disposal of sulfurcontaminated sludge on land may promote the acidification of soil. 3.1 %) than in the control (7.5 % (Ishigaki et al. and Pb from sludge is depicted in Fig.0 g/L sulfur concentration. indicating that the reduction of pH was fast with an increased concentration of inoculum. Although the removal efficiency of Zn was greater in the bioleaching sample (40.5 % after 8 days of bioleaching. 10 %. Cu.8 % Cr were removed within 12 days of bioleaching. 30°C. and Cr was 95. This was because a higher concentration of inoculum accelerated sludge acidification. the removal efficiency in the control sample was reasonably high (83.5 %.1 % to 96. 7a).4 Effort of initial inoculum concentration on bioleaching As described in Eqs. b Cu. It can be seen that bioleaching of Zn and Cu from sewage sludge was not strongly influenced by inoculum concentration with an exception of 2 % inoculum concentration. 7 under the optimized conditions. At the sulfur concentration of 5 gL−1. A. initial pH. 2 and 3. 5 gL−1 sulfur was recommended.5 g/L 2 g/L 5 g/L 10 g/L 15 g/L 50 40 30 20 10 10 0 0 2 4 6 Time (d) acidic condition (Chen and Lin 2000).7 %).e. 41. This is because Zn binding to sludge (i. the removal efficiency of Pb and Cr increased with increasing the initial inoculum concentration. and a low final pH of the sewage sludge indicate effective microbial sulfur oxidation and hence ability to leach heavy metals (Ryu et al.6–26. Cu. The removal efficiency of Pb and Cr were about 6. 37. Similar results were also observed on the bioleaching of Zn and Cu from municipal waste incineration fly ash by using sulfur-oxidizing bacteria. thiooxidans can directly oxidize metal sulfides into soluble metal sulfate. where a high rate of acidification. respectively.5 % Cu.8 % Pb. and d Cr during the bioleaching process as affected by sulphur concentrations (sludge concentration. Cr. Pb.5 % after 12 days of bioleaching when the inoculum concentration was in the range of 2–20 % (v/v).

7.t control 30 20 10 10 0 0 0 2 4 6 Time (d) 8 10 12 0 2 4 6 Time (d) 12 . 30°C. Pb. 10 %. and Cr. agitation speed. and d Cr during the bioleaching process (sludge concentration.J Soils Sediments (2012) 12:900–908 100 100 a Removal of Cu (%) 80 Removal of Zn (%) Fig. b).66. initial pH. 7 The removal efficiency of a Zn.66. 6 Effect of inoculum level on solubilization of a Zn.t control Removal of Cr (%) Removal of Pb (%) 12 50 5 Fig. sulfur concentration. indicating that the inoculation was an essential requirement a A. 150 rpm) 907 60 2% 5% 10% 15% 20% 40 20 0 0 2 4 6 8 10 12 b 80 60 2% 5% 10% 15% 20% 40 20 0 14 0 2 4 6 Time (d) 35 d Removal of Cr (%) Removal of Pb (%) 25 2% 5% 10% 15% 20% 20 15 10 40 30 2% 5% 10% 15% 20% 20 10 0 2 4 6 8 10 0 12 0 2 4 6 Time (d) exchangeable. 2 %. temperature. temperature. 12 100 b 80 80 Removal of Cu (%) Removal of Zn (%) 10 In the case of Cu.t control 60 40 20 0 2 4 6 8 10 0 12 0 2 4 Time (d) 6 8 10 8 10 12 Time (d) 50 50 d c 40 40 A. 5 gL−1. sulfur concentration. b Cu. thiooxidans can be governed by both direct and indirect mechanisms. carbonate. and Fe/Mn oxides fractions) was not strong and hence easily exchanged (Chen and Lin 2004a. agitation speed. 5 g L−1.t control 60 40 20 0 8 Time (d) 100 A. 30°C. b Cu. It is concluded that Zn bioleaching by A. 2 %. inoculum concentration. it can be seen that removal efficiency was remarkably different between the bioleached sample and the control sample after 2 days of bioleaching. c Pb. c Pb. and d Cr in sewage sludge by bioleaching (sludge concentration. 7. initial pH. 150 rpm) 10 60 c 30 0 8 Time (d) 30 20 A.

3 % Pb. Chen ZL (2010) Biodegradation characteristics of naphthalene by strain bacillus fusiformis. and 41. Lee IS. Fujita M (2005) Bioleaching of metal from municipal waste incineration fly ash using a mixed culture of sulfur-oxidizing and iron-oxidizing bacteria. These results provide the evidence that the bioleaching of Cu. Cu. Cho KS.” References Babel S. Zhang GM. Zhou M. J Air Waste Mgmt Assoc 52:237–243 J Soils Sediments (2012) 12:900–908 Couillard D. Lin JG (2004b) Bioleaching of heavy metals from livestock sludge by indigenous sulfur-oxidizing bacteria: effects of sludge solids concentration. leaching microorganisms.0 % Cr in bioleached samples were removed after bioleaching for 12 days. while only 17. thiooxidans can be used in the bioleaching of heavy metals from sewage sludge. Lin JG (2001) Bioleaching of heavy metals from sediment: significance of pH. Waste Manag 26:988–1004 Chartier M. J Hazard Mater 182:771–777 Liu YG. and Zn is mainly dominated by bioleaching indirect mechanism. Water Air Soil Poll 96:249–267 Chen SY. Chen SF. Ike M. Pb. Dacera DDM (2006) Heavy metal removal from contaminated sludge for land application: a review. percentage inoculum and nutrient enrichment on the solubilization of sediment bound metals. Nagendran R (2008) Changes in nutrient profile of soil subjected to bioleaching for removal of heavy metals using Acidithiobacillus thiooxidans. Zeng GM. and Cr because these metals strongly bind with the organic matter. and Zn from the sewage sludge was 43. This study also demonstrates that using A. Our results indicate that the bioleaching of Zn was governed by both direct and indirect mechanisms. Biores Technol 99:4124–4129 Lo KSL.6 %. Our systematic studies show that solid concentration. Moon HS. J Chem Technol Biotechnol 75:649–656 Chen SY. pH.0 % Cu. J Hazard Mater 156:102–107 Lin C. Water Res 37:2449–2457 Tyagi RD (1992) Microbial leaching of metals from municipal sludge: effects of sludge solids concentration. 95. Lin JG (2004a) Bioleaching of heavy metals from contaminated sediment by indigenous sulfur-oxidizing bacteria in an air-lift bioreactor: effects of sulfur concentration. Mozeto AA (2001) Bioleaching of metals from anaerobic sewage sludge: effects of total solids. Prasad D. Chen YH (1990) Extracting heavy metals from municipal and industrial sludges. resulting in low pH and high ORP to facilitate the heavy metals leaching from sludge. A. Can J Civ Eng 18:237–243 Hsu CH. Sreekrishnan TR (1995) Operational strategy for metal bioleaching based on pH measurements.5 %. and Cr was mainly dominated by the bioleaching mechanism. Cu and Ti in an anaerobic sewage sludge affectuated by Thiobacillus ferrooxidans and its effect on metal partitioning. Nakanishi A. Tateda M. Water Res 27:1227–1235 Du YG. Lohaza WB (1991) Fate of indicator microorganism in sludge during bacterial leaching of metals. Zhou M. Wang X (2008) Bioleaching of heavy metals from mine tailings by indigenous sulfur-oxidizing bacteria: effects of substrate concentration.908 for better acclimatization and growth of A. Garcia O Jr. under the optimized conditions. thiooxidans can utilize elemental sulfur as an energy source. respectively. Chan LC. Wong JWC (2000) Removal of heavy metals from anaerobically digested sewage sludge by isolated indigenous ironoxidizing bacteria.7 % Pb. Zn. Zeng GM. J Hazard Mater 141:202–208 Liu YG. Xu WH.2 %. Water Res 38:3205–3214 Chen SY. This suggests that A. J Environ Manage 90:2343–2353 Ryu HW. Sci Total Environ 90:99–116 Lombardi AT. Chemosphere 41:283–287 Zhang PY. Pb. carbonate. Garcia O Jr (2006) Effect of anaerobic digestion and initial pH on metal bioleaching from sewage sludge. and Fe/Mn oxide in sludge (Kumar and Nagendran 2007). For example. Biores Technol 45:33–41 Kumar RN. Ryu HW. J Environ Qual 32:751–759 Suzuki I (2001) Microbial leaching of metals from sulfide minerals. Tyagi RD (1993) Factors affecting toxic metals removal from digested sludge by enriched sulfuroxidizing microorganisms. Biores Technol 100:1394–1398 . sulfur concentration. Harrison RG (1995) Bacterial leaching of zinc and copper from mining wastes. Gan L. and 10. Biotechnol Adv 19:119–132 Tsai LJ. The bioleaching efficiency of Cr. Lee EY. Chemosphere 66:1775–1781 Kumar RN. Chemosphere 44:1093–1102 Chen SY. Choi HM (2002) Effect of solids concentration on bacterial leaching of heavy metals from sewage sludge. Fan T (2007) Effect of solids concentration on the removal of heavy metals from mine railings via bioleaching.6 %. thiooxidans is a potential technique to bioleach heavy metals from sewage sludge. Chemosphere 60:1087–1094 Jain DK. Hydrometall 37:169–179 Ishigaki T. Pb. 9. Acknowledgments The authors sincerely thank Fujian Normal University for financial support through the “Min-Jiang Fellowship.6 % Cu. J Environ Eng 121:527–535 Fang D. Lin JG (2000) Influence of solid content on bioleaching of heavy meals from contaminated sediment by Thiobacillus spp. thiooxidans (Kumar and Nagendran 2007). Al. Zhou LX (2007) Enhanced Cr bioleaching efficiency from tannery sludge with coinoculation of Acidithiobacillus thiooxidans TS6 and Brettanomyces B65 in an air-lift reactor. 43. Nagendran R (2007) Influence of initial pH on bioleaching of heavy metals from contaminated soil employing indigenous Acidithiobacillus thiooxidans. and inoculum concentration effectively influenced the bioleaching of metals. while the bioleaching of Cu. Dastidar MG. 96. Zhu Y. Sreekrishnan TR (2009) Bioleaching of heavy metals from sewage sludge: a review. and 96. Tyagi RD. Choi H (2003) Leaching characteristics of heavy metals from sewage sludge by Acidithiobacillus thiooxidans MET. Process Biochem 27:89–96 Villar LD. J Environ Sci Health A 36:793–806 Pathak A. 41. Chemosphere 69:303–310 Henry JG. Zeng GM. Li X. Yu KC. 4 Conclusions The present study demonstrated that the isolated indigenous sulfur-oxidizing bacteria A.4 % Cr were removed in control sludge medium. Garcia O Jr (2002) Biological leaching of Mn. Kung PY (2003) Effect of temperature on removal of heavy metals from contaminated river sediments via bioleaching. Water Res 36:3193–3202 Lombardi AT. thiooxidans plays an important role in the bioleaching of Cu. Couillard D (1997) Biological processes: the effects of initial pH. and energy source. Mercier G (1993) Removal of metals and fate of N and P in the bacterial leaching of aerobically digested sewage sludge. Zou S. Pb. J Environ Sci Health A 41:211–222 Xiang L. Wu Z (2009) Sewage sludge bioleaching by indigenous sulfur-oxidizing bacteria: effects of ratio of substrate dosage to solid content. Chemosphere 54:283–289 Cho KS.

& Remediation is the property of Springer Science & Business Media B. . users may print. However. Risk Assessment. or email articles for individual use. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission.V.Copyright of Journal of Soils & Sediments: Protection. download.