Minerals Engineering 98 (2016) 279–285

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Minerals Engineering
journal homepage: www.elsevier.com/locate/mineng

The effect of potential heap construction methods on column

bioleaching of copper flotation tailings containing high levels
of fines by mixed cultures
Xiaodong Hao, Yili Liang ⇑, Huaqun Yin, Liyuan Ma, Yunhua Xiao, Yazi Liu, Guanzhou Qiu, Xueduan Liu ⇑
School of Minerals Processing and Bioengineering, Central South University, Changsha, China
Key Laboratory of Biometallurgy, Ministry of Education, Changsha, China

a r t i c l e i n f o a b s t r a c t

Article history: It is known that excess fines may reduce heap permeability and block channelings of leachate flow in
Received 12 January 2016 heap bioleaching operation, and further cause low metal recovery. The purpose of this investigation
Revised 2 June 2016 was to compare the effects of three potential heap construction methods including layered heap con-
Accepted 27 July 2016
struction method (Method A), agglomerate heap construction method (Method B) and pelletized sinter-
ing heap construction method (Method C) of copper flotation tailings on column bioleaching behaviors. In
the three heap construction methods, the tailings copper extractions achieved 54.61%, 60.09% and 43.93%,
Keywords:
respectively, in Method A, B and C on day 83. Copper extraction of Method B reached maximum. In addi-
Fines
Heap construction methods
tion, compositions and structures of microbial communities were examined using Illumina Miseq
Copper flotation tailings sequencing technology based on 16S rRNA amplification. Acidithiobacillus, Leptospirillum and
Column bioleaching Ferroplasma were main microorganisms in three heap construction methods. Detrended correspondence
Microbial community analysis showed that Method C had little effect on microbial communities. These studies revealed the
influence of different heap construction methods on leaching behaviors and microbial dynamics, which
will facilitate the bioleaching of fine-grained ores.
Ó 2016 Published by Elsevier Ltd.

1. Introduction heap. The impermeable layers would affect the transfer of oxygen
and carbon dioxide and eventually result in low metal recovery
Due to the continuous exploitation and consumption of efficiency. Thus, heap construction methods and raw ores pretreat-
valuable mineral resources, metal recovery from the tailings and ment were the principal factors in bio-heap construction.
low-grade ores becomes increasingly important. Traditional Many methods were utilized to resolve this problem. Garcia and
pyrometallurgy grows less desirable for tailings or low-grade ores Jorgensen (1997) had indicated that the ores should be determined
on account of the stringent requirement of environmental protec- for agglomeration supposing that the percentage of fines smaller
tion and the inefficient use of resources and energies (Brierley, than 75 lm was >15%. Many studies had applied laboratorial prac-
2008; Watling, 2006). However, bioleaching, as an emerging and tice of agglomeration with many polymeric binders such as poly-
frontier technology, has been applied into the extraction of copper, acrylamide, stucco, bentonite et al. into metal recovery in heap
cobalt, nickel, zinc and uranium from low grade sulfide minerals. leaching operations and gained high metal leaching rates
The megaton flotation tailings used in this study were from (Bouffard, 2008; Kodali et al., 2011; Lewandowski and Kawatra,
Lualo in Zambia. It has high fines content (the proportion was 2009). In the 1980s, Sociedad Mineral Pudahuel originally applied
48% of <75 lm), but a low clay content. As the leachate percolates the thin-layer leaching concept into copper oxides and copper sul-
through the heap containing high levels of fines, the raffinate can fides extractions in the copper industry, which contributed to the
cause migration of fine particles and clog the natural flow channels remarkable expansion of the heap bioleaching, solvent extraction,
within the ore bed, which form impermeable layers inside the and electrowinning process in South America. Although above
methods had advanced the industrial practice of copper heap
leaching operations, the effects of heap construction method and
⇑ Corresponding author at: School of Minerals Processing and Bioengineering,
Central South University, Changsha, China.
ores pretreatment on metal recovery were still unclear.
E-mail addresses: [email protected] (Y. Liang), [email protected] Understanding the microbial structure and succession was also
(X. Liu). important to advance commercial bioheap leaching operations

http://dx.doi.org/10.1016/j.mineng.2016.07.015
0892-6875/Ó 2016 Published by Elsevier Ltd.
280 X. Hao et al. / Minerals Engineering 98 (2016) 279–285

(Brierley, 2001; Rawlings, 2002). Brierley and Brierley (2013)

showed that potential interactions among microbial community
dynamics were the keys to improve bioleaching and biooxidation.
In most cases, the investigation of microbial populations was
mainly focused on free microorganisms in the leachate. However,
attached microorganisms on the mineral surfaces also played
undeniable roles on bioleaching (Gautier et al., 2008). Therefore,
it was necessary to monitor the shifts of attached microorganisms
on the mineral surfaces during tailings bioleaching. Although many
nucleic acid-based molecular methods (e.g., RFLP, SSCP, DGGE) had
been applied to determine microbial structures in bioleaching
environments, high-throughput and high-resolution detection
methods could offer a rapid and simple method to quantify the
microbial communities (Orgiazzi et al., 2013).
Therefore, in this paper, three potential heap construction
methods were carried out to investigate the column bioleaching
of fine-grained flotation tailings by mixed cultures. This paper
attempts to detail the effects of heap construction methods and
raw ore pretreatments on leaching behaviors and microbial
dynamics of fines bioleaching, and the data presented in this study
would be helpful in the industrial applications of copper industry.

2. Materials and methods

2.1. Minerals components Fig. 1. Diagram of the column bioreactor in three potential heap construction
methods of flotation tailings. 1, glass container; 2, peristaltic pump; 3, sterile
The copper flotation tailings in this test were obtained from tail- sponge; 4, flotation tailings; 5, quartz sand; 6, Yulong mineral; 7, support plate.

ings dam in Lualo, Zambia. The percentages of different particle

size distributions of tailings were 48% (<75 lm), 43% (75–
2.2.3. Pelletized sintering heap construction method (Method C)
150 lm) and 9% (150–270 lm), respectively. Copper phase analy-
The tailings added with 1.5% (w/w) bentonit were placed into
sis showed 29.47% free copper oxide, 6.32% combined copper
an agglomeration drum. The drum was turned at 20 rpm with 5°
oxide, 28.95% secondary copper sulfide and 35.26% primary copper
inclination while deionized water as binder was sprayed onto the
sulfide. The main chemical composition of tailings was (w/w)
ores for 20 min. The deionized water was applied with a spray bot-
0.19% Cu, 19.73% Fe, 2.25% S, 3.27% Ca, 1.25% Na, 4.6% Al and
tle at a dosage of 0.15 mL/g ores. After the agglomeration process
1.4% K. Whole ore substrate used in this study was obtained from
completed, the agglomerates were removed from the drum and
the Yulong copper mine in Xizang of China, which contains (w/w)
passed through the 6–8 mm screen. The agglomerates were air
2.75% Cu, 16.64% Fe, 17.41% S and 10.47% Al.
dried at 60 °C for 4 h, then placed into the muffle remaining
800 °C for 10 min. Parts of agglomerates were cooling and sub-
2.2. Column bioreactor configuration and heap construction methods mersed completely into a H2SO4 solution (pH 1.5) for 24 h to test
the integrity of agglomerates.
50 cm high column made of 0.5 cm thickness plexiglass with In Method B and C, a layer of support quartz sand sized at 2.5–
the internal diameter of 7 cm were used in this study. A plexiglass 5 mm was placed 20 mm thickness at the bottom of the columns.
support plate with numerous 0.3 cm holes was placed at the bot- In each heap construction method, approximate 1.3 kg weight tail-
tom of the column. The leaching solution passed through the ore ings were loaded into columns. Finally, a piece of sterile sponge
samples by gravity and recirculated through a side loop with a was placed on the top surface of each column to ensure the uni-
peristaltic pump (Fig. 1). form wetting of the ores (Fig. 1).

2.2.1. Layered heap construction method (Method A) 2.3. Mixed microorganisms culture
The tailings were mixed with 10% (v/v) H2SO4 solution ade-
quately, and the liquid ratio (v/w) was 20%. 5 cm thickness quartz Pregnant leaching solution (PLS) and leaching slags were
sand sized at 2.5–5 mm was loaded at the bottom of the column, collected from the Dexing Copper Mine in Jiangxi, China. The
then 5 cm thickness tailings mixed with H2SO4 solution were microorganisms in PLS were harvested by centrifuging at
placed on the surface of the quartz sand. Then quartz sand and tail- 12,000 rpm for 15 min. The leaching slags were washed with pH
ings were loaded in accordance with this order. Finally, four alter- 2.0 sterilized deionized water, and the supernatant was
nating layers of quartz sand and tailings were used in the column, centrifuged as well as PLS. The microorganisms from PLS and slag
respectively (Fig. 1). surfaces were gained and then inoculated into the pH 2.0 culture
medium at 30 °C for the enrichment of initial inoculum. The
composition of culture medium was 3 g/L (NH4)2SO4; 0.5 g/L
2.2.2. Agglomerate heap construction method (Method B) MgSO4
7H2O; 0.1 g/L KCl; 0.5 g/L K2HPO4; 0.01 g/L Ca(NO3)2. In
The tailings blended fully with Yulong mineral sized at 2.5– addition, FeSO4
7H2O (44.7 g/L), S (10 g/L) and tailings (2%, w/v)
5 mm, and the ratio of tailings and Yulong mineral was 2:1 (w/ were added as energy sources. Finally, microorganisms were
w). The mixed ores were mixed with 10% (v/v) H2SO4 solution harvested when the cell density reached 1  108 to 2  108
and the liquid ratio (v/w) was 20% as well. Then agglomerate ores cells/mL and then washed with pH 2.0 culture medium, which
were loaded into the column (Fig. 1). resulted in the initial inoculum.
 X. Hao et al. / Minerals Engineering 98 (2016) 279–285 281

2.4. Bioleaching tests was at 72 °C for 10 min. After each sample was amplified in tripli-
cate following the thermal cycling, the PCR products were sepa-
In this study, the solution permeability experiment was carried rated by 1.0% agarose gel electrophoresis and visualized by
out to determine the spray rate of leachate. When the spray rate staining with ethidium bromide under UV light. Replicate PCR
was about 0.66 L/h, the solution caused the saturation of the ores products for each sample were pooled and purified using E.Z.N.A.
in Method A. While the spray rate was approximately 0.90 L/h, Gel Extraction Kit (437 bp) (Omega Bio-Tek, Inc., USA). A single
the solution could lead to the saturation in Method B but not for sample for sequencing was prepared by combining approximately
Method C. Hence, the solution permeability order in three heap equimolar amounts of PCR products. Sequencing process was car-
construction methods was regarded as Method C > Method ried out on Illumina Miseq sequencing platform. The Miseq 500
B > Method A. And each bioreactor was fed with leaching solutions cycles kit was used for 2  250 bp paired-ends sequencing on
at the rate of 0.6 L/h, which did not cause the saturation of ores. Miseq machine.
The column bioleaching experiments were tested at ambient
temperature. The leaching process was divided into two stages
2.7. Processing of Illumina sequencing data
including acid pre-leaching stage (10 days) and bioleaching stage
(73 days). The ores were pre-leached with 1.6 L culture medium
Raw data generated from the Illumina Miseq sequencing run
(pH 1.5) in order to neutralize the acid-consuming minerals. The
were processed and analyzed following the Galaxy pipeline on
pH was adjusted using concentrated sulfuric acid everyday in the
our local server (Pond et al., 2009). Quality reads were assigned
pre-leaching stage, but not during the bioleaching stage. PLS evap-
to samples subsequently in accordance with their unique barcodes,
oration was supplemented by adding sterilized deionized water to
and were trimmed using Btrim with threshold of QC higher than 20
maintain constant volume. After the tests ended, the columns were
over five bp window size and the minimum length of 100 bp
unloaded and the residues were acid washed, dried and prepared
(Caporaso et al., 2012). Forward and reverse files with at least
for the final analysis.
10 bp overlap length and lower than 5% mismatches were joined
using Flash (Magoč and Salzberg, 2011). OTU clustering was
2.5. Analysis methods
through UPARSE using average clustering algorithm at 97% similar-
ity level (Edgar, 2010), and taxonomic assignment was through
The composition of the mineral samples was analyzed by Induc-
RDP classifier (Wang et al., 2007) with a minimal 50% confidence
tively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES).
estimate.
Samples of leaching solution (2 mL) were regularly collected for
measurement of pH and the concentrations of copper and iron.
Copper and total iron concentrations were analyzed by ICP-AES. 3. Results and discussion
The concentration of ferrous iron was determined by phenanthro-
line colorimetry, while ferric iron concentration was the difference 3.1. Effects of potential heap construction methods on bioleaching
between the concentrations of total iron and ferrous iron. The pH behavior
in PLS was measured with a pH meter (pHS-3C, Leici, China). The
cell density in leachate was monitored by direct counting using a The column bioleaching of copper flotation tailings by mixed
hemocytometer with an optical microscope. After the experiments, cultures was carried out in three heap construction methods for
the tailings residues in Method B were separated from the mixed 83 days. As shown in Fig. 2a, in Method A and B, the copper con-
ores by passing them through a 270 lm screen, which was washed centrations increased slowly, then remained stable in the late
by flowing water. Then, the tailings residues were collected, dried bioleaching stage. The addition of Yulong mineral in Method B
and calculated copper extraction. resulted in a higher copper concentration than Method A and C.
In Method C, the copper concentration increased rapidly in acid
2.6. DNA extraction and PCR amplification leaching stage, while accelerated slightly during bioleaching. This
may be attributed to the reason that the tailings did not be cured
The genomic DNA from initial inoculum and mixed microorgan- with H2SO4 before agglomeration. Fig. 2b showed that the copper
isms in PLS on day 83 were extracted using a TIANamp Bacteria extractions of tailings based on copper contents were 54.61%,
DNA kit (Tiangen Biotech, Co., Ltd., China) according to the manu- 60.09% and 43.93%, respectively, in Method A, B and C on day 83.
facturer’s instruction. The total DNA was extracted from leached Method TB represents the mixed minerals copper extraction
residues at the top and bottom of the columns respectively as including tailings and Yulong mineral in Method B, which was up
described previously (Zhou et al., 1996). Meanwhile, the strong to 81.12%.
and weak attached microorganisms were also separated from Fig. 3 showed the variation of pH in leachate during column
leaching residues of the whole columns. Leaching residues (10 g) bioleaching in three heap construction methods. The pH increased
were loaded into 500 mL-glass flasks including 100 mL-culture in bioleaching stage, which was up to 2.2, 2.3 and 2.5 on the 13th
medium (pH 2.0) and then shaked at 175 rpm for 5 min at room day in Method A, B and C, respectively. Thereafter, it stabilized
temperature. The genomic DNA of the weak and the strong until the 25th day. Finally, the pH values declined to 1.6, 1.3 and
attached microorganisms were extracted from the supernatant 2.35, respectively at the end of bioleaching. The rising pH during
and the residues, respectively. the first 10 days may be due to the ferrous iron oxidation (con-
Two primers, F515 (50 -GTGCCAGCMGCCGCGGTAA-30 ) and R806 sumed proton), which was in accordance with the previous result
(50 -GGACTACHVGGGTWTCTAAT-30 ) with Illumina adapter (Wang et al., 2014). The lower pH in Method B may be attributed
sequences (Caporaso et al., 2012), were used to amplify the V4 to the addition of Yulong minerals (17.41% S), and more elemental
hypervariable region of bacterial and archaeal 16S rRNA gene frag- sulfur was released and oxidized by sulfur-oxidizers. In bioleaching
ments (Bates et al., 2011). PCR system (25 ll) consisted of 1 ll process, elemental sulfur would be released through the minerals
(10 lM) of each forward and reverse primer, 1 ll of template oxidation after the attack of ferric iron.
DNA, 12.5 ll of 2  Taq PCR Master Mix (Tiangen Biotech, Co., In three potential heap construction methods, the total iron
Ltd., China) and 9.5 ll ddH2O. Samples were initially denatured concentration variation presented different trends during column
at 94 °C for 5 min, then amplified by using 30 cycles of 94 °C for bioleaching process (Fig. 4). In the acid leaching stage, the ferrous
45 s, 62 °C for 45 s, and 72 °C for 1 min. A final extension process iron concentration was equal to the total iron concentration, and
282 X. Hao et al. / Minerals Engineering 98 (2016) 279–285

Fig. 2. Variation of copper concentrations in leachate and copper extractions based on copper contents of residues in three potential heap construction methods. (a) The
copper concentration; (b) Copper extraction. Method A, B and C in (b) represent the tailings copper extractions; Method TB represents the mixed minerals copper extraction
containing tailings and Yulong mineral.

reached the lowest values of 1.16  107, 1.6  106 and 0.8  106
cells/mL in Method A (day 25), B (day 17) and C (day 21), respec-
tively. It may be due to that the lower permeability caused the
longer time of microorganisms adsorbed on the ore surfaces. The
addition of Yulong mineral containing 17.41% S and 16.64% Fe
could provide more energy sources to promote the growth of
microorganisms in Method B. The cell density decreased in the
final stage, and it may result from the decrease of pH and the accu-
mulation of heavy metal ions and organic substances, which
severely inhibited the growth and activity of the microorganisms.
In Method C, the cell density remained the lowest among three
heap construction methods during the whole leaching process,
and the reason was mainly the sulfur loss of tailings in sintering
process and high pH value in leachate, which could affect the
growth of acidophilic microorganisms.

3.2. Microbial community dynamics analysis of free and attached

Fig. 3. Variation of pH in leachate during column bioleaching of tailings in three microorganisms
potential heap construction methods.
Bioleaching is a complex process and affected by many biolog-
ical, chemical and physical factors. These factors will determine the
only low ferric iron was present. However, the ferric iron concen- succession of the microbial community structures in different
tration generated from ferrous iron oxidation by mixed cultures depth of the bioheap (Pradhan et al., 2008). The structures of
increased rapidly, while ferrous iron concentration stayed low. bacteria and archaea in three heap construction methods during
Total iron concentration reached 2.63 g/L on the 13th day and column bioleaching of tailings were detected by Illumina Miseq
remained stable during bioleaching in Method A. Compared with sequencing platform based on 16S rRNA PCR amplification. The
Method A and C, there was higher total iron concentration and cop- microbial community structures and compositions from the PLS
per extraction in Method B. It was demonstrated that high concen- (PLS-FREE), leaching residues at the top (LS-TOP) and bottom
tration of total iron was beneficial to copper extraction (Yu et al., (LS-BOTTOM) of the columns and the strong (LS-STRONG) and
2014). It was worth noting that the total iron concentration weak (LS-WEAK) attached microorganisms in the whole columns
decreased from 1.44 g/L on day 13 to 0.15 g/L on day 83 in Method after experiments ended were analyzed shown in Fig. 6.
C. It was inferred that higher pH in leachate greatly promoted the The results indicated that Acidithiobacillus (67.80%), Sulfobacillus
formation of jarosite, and drastically reduced the available oxidiz- (24.64%), Leptospirillum (2.93%) and the only archaea genera Ferro-
ing reagent (ferric ion) in leachate (Ojumu and Petersen, 2011). plasma (1.62%) existed in the original inoculum (Fig. 6). In three
These jarosite particles aggregated together to form a layer to wrap heap construction methods, Acidithiobacillus, Leptospirillum and
the mineral surfaces, especially in the leaching reaction place, Ferroplasma were predominanted. In addition, Acidiphilium was
inhibited the interaction between ores and microorganisms, which also the main organism of Method C. However, Sulfobacillus was
resulted in a lower copper extraction in Method C. not the dominant microorganism in three heap construction meth-
The variation of free cell density in leachate was shown in Fig. 5. ods during bioleaching.
The initial cell density was 4.7  107 cells/mL. The number of free The microbial community population of Method A in five sam-
cells decreased with the increase of permeability in three heap ples presented large differences. Acidithiobacillus and Leptospirillum
construction methods during bioleaching process. In the early were the dominant species in PLS, accounting for approximately
stage of bioleaching, the density of free cells kept decreased and 35.73% and 50.23%, respectively. Besides, Leptospirillum had a
 X. Hao et al. / Minerals Engineering 98 (2016) 279–285 283

Fig. 4. Variation of iron ion concentration in leachate during column bioleaching of tailings in three potential heap construction methods. (a) Method A; (b) Method B; (c)
Method C.

additional air was not forced into the column bioreactors. Oxygen
should be consumed by the oxidation of sulfide minerals, ferrous
ion, RISCs or energy yielding reactions (Acevedo and Gentina,
1989), which resulted in the decrease of oxygen concentration at
the bottom of the column. As a result, these autotrophic strains
tended to be restrained by the consumed oxygen and carbon diox-
ide at the bottom. In addition, Pseudomonas was also detected in
LS-TOP, LS-BOTTOM and LS-WEAK. In the final bioleaching stage,
a high concentration of dissolved carbon which derived from
microorganisms as exudates, cell lysates and minerals dissolving
(Okibe et al., 2003) could stimulate the growth of heterotrophic
microorganisms (Pseudomonas and Acidiphilium). Previous studies
indicated that Pseudomonas not only oxidized or reduced elemen-
tal sulfur and other sulfides (Patel et al., 2009; Rzhepishevska,
2008), but also promoted the dissolution of minerals through the
secretion of organic acids such as citric acid, oxalic acid and suc-
cinic acid.
Fig. 5. Variation of free cell density in leachate during column bioleaching of In Method B, the microbial community in PLS-FREE had a great
tailings in three potential heap construction methods. difference from other four samples. Acidiphilium (23.86%) existed
in PLS, but not detected in leaching residues. Compared with the
original inoculum, the proportion of Ferroplasma increased from
higher abundance in PLS than other four samples. Previous study 1.62% to 18.42%, 84.66%, 66.12%, 88.78% and 22.04% in five sam-
had indicated that Acidithiobacillus and Leptospirillum frequently ples, respectively. Previous reports had indicated that Ferroplasma
existed in the most bioleaching systems. It was interesting that a often grows well in the final bioleaching stage (Dopson et al., 2004;
lower ratio of these strains in LS-BOTTOM was found probably Hawkes et al., 2006; Zhang et al., 2009). The PLS became increas-
due to air depletion from the top to the bottom. In this work, ingly acidic (Fig. 3) and enriched with high metal concentrations
284 X. Hao et al. / Minerals Engineering 98 (2016) 279–285

Fig. 6. The analysis of microbial community structures from initial inoculum, PLS and leaching residues at different positions of columns on day 83. The pie chart represents
the initial inoculum.

and organic matters (Okibe et al., 2003), which favored the growth
of the archaeon Ferroplasma (Zhou et al., 2008). However, other
bacteria might be disintegrated, and to some extent their cellular
activities were inhibited under the metabolic and metallic stresses
in the final stage. In addition, the average relative abundance of
Acidithiobacillus of Method B was lower than that of Method A
and C since the optimal pH for Acidithiobacillus was about 2.0.
The microbial community structure of Method C was signifi-
cantly different from that of Method A and B. Acidiphilium was
not detected in the original inoculum, but became abundant in
the later stage in Method C, accounting for approximately
25.62%, 16.80%, 12.86%, 11.32% and 7.09% in five samples, respec-
tively. Acidiphilium had a wide range of pH tolerance, but the opti-
mal pH was above 2.0. Previous studies had demonstrated that
Acidiphilium not only acted as a consumer of organic matters pro-
duced by autotrophic acidophiles like Acidithiobacillus sp., Lep-
tospirillum sp. and Sulfobacillus sp. (Johnson and Hallberg, 2003;
Zeng et al., 2010), but also had significant leaching capacity Fig. 7. DCA analysis of microbial community structures based on OTU data. H, N,
and represent the samples of inoculum, Method A, B and C, respectively; I, P, T, B,
towards sulfide ores (Zhang et al., 2013). In addition, Sulfobacillus
W and S represent microbial community structures of the inoculum, PLS-FREE,
existed in all five samples. It could utilize both ferrous iron and sul- LS-TOP, LS-BOTTOM, LS-WEAK and LS-STRONG in three heap construction methods,
fur as energy sources. The optimal pH was 2.4–2.5 when oxidizing respectively.
sulfur and 1.5–1.7 when utilizing ferrous iron (Yu et al., 2014). The
appropriate pH of PLS in Method C was beneficial to sulfur oxida-
tion of Sulfobacillus. However, low sulfur contents (2.25% S) inhib- LS-TOP, LS-WEAK and LS-STRONG samples. In addition, the
ited the growth of Sulfobacillus. percentage of Ferroplasma reached maximums on the mineral
Detrended Correspondence Analysis (DCA) in Fig. 7 presented surfaces in LS-TOP, LS-BOTTOM and LS-WEAK samples of Method
the potential correlation of microbial community across different B, which made a distinction with the situation in PLS-FREE and
samples in columns. The results indicated DCA with 75.88% of LS-STRONG. Interestingly, the spot distributions of Method C
the total variance explained by the first axis (DCA 1, 48.26%) and were clustered together due to the relative abundance of dominant
the second axis (DCA 2, 27.62%). The initial inoculum had a greater and rare species were similar in five samples (see Fig. 6). The
distance from other samples indicating that the microbial commu- microbial populations of Method A, B and C were different on
nity structures changed greatly in the process of bioleaching. In account of the mineral processings and heap construction
inoculum, Sulfobacillus was over 20% as the predominant microor- methods. The uneven distribution of the gas under different heap
ganisms, but Leptospirillum and Ferroplasma just accounted for construction methods and pH variation in the final bioleaching
2.93% and 1.62%, respectively shown in Fig. 6. However, after stage may lead to the discrepancy of microbial community
bioleaching Leptospirillum and Ferroplasma became the dominant structures in different positions of column.
species, but Sulfobacillus was hardly detected in three heap con-
struction methods, which had the difference from the microbial 3.3. Operation feasibility
community structure of initial inoculum. In Method A, proportion
of Leptospirillum was more than that of Acidithiobacillus and It is thought that the three potential heap construction methods
Ferroplasma in PLS-FREE, while Leptospirillum gained a competitive of flotation tailings in this study could be employed in constructed
disadvantage compared with Acidithiobacillus and Ferroplasma in heaps or large columns. Layered heap construction method
 X. Hao et al. / Minerals Engineering 98 (2016) 279–285 285

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This work was financially supported by the National Basic
of microbial community under the adjustment of initial and processing pH
Research Program of China (2010CB630901), the National High during bioleaching of chalcopyrite concentrate by moderate thermophiles.
Technology Research and Development Program of China Bioresour. Technol. 162, 300–307.
(2012AA061502) and the National Nature Science Foundation of Zeng, W., Qiu, G., Zhou, H., Peng, J., Chen, M., Tan, S.N., Chao, W., Liu, X., Zhang, Y.,
2010. Community structure and dynamics of the free and attached
China (41573072 and 31570113). microorganisms during moderately thermophilic bioleaching of chalcopyrite
concentrate. Bioresour. Technol. 101, 7068–7075.
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