Minerals Engineering 180 (2022) 107514

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

Stirred-tank bioleaching of copper and cobalt from mine tailings in Chile

Ruiyong Zhang 1, Axel Schippers *
Federal Institute for Geosciences and Natural Resources (BGR), Geomicrobiology, Stilleweg 2, 30655 Hannover, Germany

A R T I C L E I N F O A B S T R A C T

Keywords: Metals from low-grade sulfide ores, electronic waste and mine tailings can be recovered using bio­
Bioleaching hydrometallurgy. In this study, acidophilic microbial consortia of mesophiles and moderate thermophiles were
Mine tailings applied in shake flasks as well as in stirred tank reactors (STR) to bioleach cobalt and copper from mine tailings
Cobalt
in Chile. The mine tailings near Taltal contained considerable amounts of copper (~5000 ppm) and cobalt
Moderate thermophiles
Stirred tank reactor
(~115 ppm). After adaptation of the mesophilic consortium, 38% of Co and 86% of Cu were recovered in 2-L STR
at 10% solid load after 12 days bioleaching at 30 ◦ C. In a mini-pilot test in 20-L STR at 15% solid load, Co and Cu
extraction reached 49% and 83%, respectively. Improved metal recovery was obtained by using a moderate
thermophilic microbial consortium at 42 ◦ C, reaching 74% of Co and almost 100 % Cu in 2-L STR at 10% solid
load. Low Fe dissolution below 14% for all tests was observed which may favor the downstream processing of Cu
and Co. Together with the results from chemical leaching, the data indicated that Cu was mainly leached by
sulfuric acid (acid leaching) while a high Co extraction required Fe(II)-oxidizing microbial activity (bioleaching).

1. Introduction recovery of Cu and Co (Ahmadi et al., 2015; Marrero et al., 2015;

Chile is the country with the highest copper reserves and has the
highest copper mining activity, which generated huge quantities of ore
processing residues known as mine tailings. These tailings contain not
extracted, partly economically relevant quantities of base metals such as
copper and cobalt, thus, they can be considered as potential metal re­
sources. Due to the increased demand of metals and the gradual deple­
tion of the conventional ore deposits, metal recovery from mine tailings
has been raised increased attention. For an economic reprocessing of
mine tailings for Cu and Co recovery a suitable process scheme for each
case needs to be developed including various steps (Alcalde et al., 2018;
Berkh et al., 2019; Kinnunen and Kaksonen, 2019; Dold, 2020; Araya
et al., 2021; Drobe et al., 2021; Schueler et al., 2021; Shengo, 2021).
Bioleaching takes advantage of acidophilic bacteria, which generate
ferric ions and sulfuric acid for dissolution of metal sulfide minerals and
metal solubilization. Biomining processes are estimated to be relatively
low-cost, environmentally friendly and suitable for both, large scale as
well as small scale applications for low-grade ores and mine residues
(Vera et al., 2013; Schippers et al., 2014; Kaksonen et al., 2018). One
processing option is bioleaching of mine tailings, which has been suc­
cessfully tested in a number of laboratory to pilot studies also for the
Falagán et al., 2017; Altinkaya et al., 2018; Mäkinen et al., 2020; Zhang
et al., 2020; Hao et al., 2021; Shengo, 2021) or even in an industrial
application (Morin and d’Hugues, 2007). One example of bioleaching of
copper from flotation tailings is the copper mine at Bor, Serbia, where
the benefits of a higher metal recovery with bioleaching compared to
acid leaching was demonstrated, combined with a lower environmental
impact (Stanković et al., 2015).
In most of the studies, mesoacidophilic iron- and sulfur-oxidizing
bacteria and sometimes also archaea were used for bioleaching of the
sulfidic tailings. The following are some bioleaching examples for Cu
and Co recovery. A maximum copper extraction of 60 % was obtained in
column bioreactor tests with copper flotation tailings (~0.2% Cu) from
Zambia (Hao et al., 2021). Bioleaching in 10-L stirred tank reactors
(STR) reached metal leaching of 87% Co and 43% Cu from Scandinavian
tailings (Mäkinen et al., 2020). The most effective extraction of copper
(84 to > 90%) was achieved by bioleaching the tailings from Bor, Serbia
and Sevilla, Spain at 45 ◦ C, using a defined thermo-tolerant microbial
consortium and addition of elemental sulfur allowing for a low pH 1
(Falagán et al., 2017). Co and Cu extraction achieved even 91% and
57%, respectively, from bulk tailings of the abandoned Rammelsberg
mine, Germany in STR bioleaching experiments, and 66% Co and 33%

* Corresponding author.
E-mail address: axel.schippers@bgr.de (A. Schippers).
1
Present addresses: Key Laboratory of Marine Environmental Corrosion and Biofouling, Institute of Oceanology, Chinese Academy of Sciences, 266071 Qingdao,
China; Open Studio for Marine Corrosion and Protection, Pilot National Laboratory for Marine Science and Technology (Qingdao), 266237 Qingdao, China.

https://doi.org/10.1016/j.mineng.2022.107514
Received 31 January 2022; Received in revised form 15 March 2022; Accepted 16 March 2022
Available online 23 March 2022
0892-6875/© 2022 Elsevier Ltd. All rights reserved.
R. Zhang and A. Schippers
Cu were extracted from a tailings flotation concentrate (Zhang et al.,
2020). STR bioleaching experiments at 40 ◦ C using pyrrhotite-rich
tailings from Sudbury, Canada showed 90–100% and 80–100% leach­
ing of Ni and Co, respectively (Cameron et al., 2018). These two metals
could also be extracted, 53–57% Ni and 55–60% Co, from non-sulfidic
tailings of laterite processing in STR bioleaching experiments supple­
mented with elemental sulfur (Marrero et al., 2015). STR processing
showed a good performance for bioleaching (Vakylabad et al., 2012).
Reprocessing flowsheets for copper mine tailings in Chile combining
flotation, magnetic separation and acid leaching were recently pub­
lished without considering bioleaching (Drobe et al., 2021). This study
explores the efficiency of bioleaching for metal extraction (i.e. Cu and
Co) from these tailings in Chile. Different acidophilic microbial con­
sortia including sulfur- and iron- oxidizing mesophiles, moderate ther­
mophiles as well as indigenous bacteria from tailings were used in
bioleaching experiments in shake flask as well as in batch 2-L and 20-L
STR to treat tailings’ samples from four sites in Chile.
2. Materials and methods
2.1. Tailings samples and characterization
Since the work was part of a collaboration of BGR (Bundesanstalt für
Geowissenschaften und Rohstoffe), the German Chamber of Commerce
in Chile (AHK Chile) and the Chilean state organization SERNAGEOMIN,
tailings of the state owned company ENAMI (Empresa Nacionál de
Minería, Santiago de Chile, Chile) were accessible and sampled from
four tailings sites (Supplementary Fig. S1; Drobe et al., 2021): Minera
Clarita (NW San Felipe, S 32◦ 41‘25‘‘ E 70◦ 47‘29‘‘), Minera Carola (SE
Copiapó S 27◦ 34‘21‘‘ E 70◦ 14‘58‘‘), Planta El Salado (E Chañaral S
26◦ 25‘16‘‘ E 70◦ 19‘06‘‘), Planta Taltal (Taltal S 25◦ 23‘53‘‘ E 70◦ 28‘23‘‘).
From each tailings dump, about 20 kg samples were collected at three to
five locations close to the surface for shake flask bioleaching tests. The
Taltal tailings were additionally sampled in more detail with 18 drilling
locations. Inside the Taltal tailings, a vertical and horizontal gradation
of the material was observed according to the deposition velocity of the
tailings particles. Two size fractions could be identified, a coarse sized
fraction was usually of black to grey colour and a fine-grained clayish
material was either black, reddish or brownish (Supplementary Fig. S1;
Drobe et al., 2021). Fine and coarse material from all Taltal coring lo­
cations was mixed each for a bulk fine and a coarse tailings sample. Both
samples were sieved and the fraction < 500 µm (99 % of the sample
material) was stored under nitrogen in gas-tight bottles to avoid
oxidation reactions. These two samples were separately used in bio­
leaching experiments in stirred-tank reactors.
The elemental composition of the two Taltal tailings samples and of
bioleaching residues was determined by peroxide fusion ICP-MS (FUS-
MS-Na2O2, Actlabs), X-ray fluorescence analysis (XRF) and chemical
analysis. For the latter, the samples were digested by aqua regia and
then analyzed for their metal ion concentration by ICP-OES.
Total sulfur and organic carbon contents were analyzed using a LECO
CS 230 carbon–sulfur analyzer. For the latter, samples were first treated
with 10% HCl at 80 ◦ C to remove inorganic carbon. Treated samples
were then combusted in a high frequency oven at about 2000 ◦ C with an
oxygen stream. The resulting carbon dioxide was analyzed by infrared
detection, carbon monoxide was catalytically oxidized to carbon dioxide
before detection. The sulfide content was determined by using
chromium-reducible sulfur (CRS) distillation as previously described
(Gröger et al., 2009). The specific surface area was determined by N2
adsorption with a Micromeritics Gemini III 2375 surface area analyzer
(Kaufhold et al., 2010). The mineralogical composition was determined
via X-ray diffraction analysis (XRD) with Rietveld refinement (data
taken from Drobe et al. 2021). Surface analyses of solids was done by
electron probe microanalysis (EPMA-SEM).
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Minerals Engineering 180 (2022) 107514
2.2. Bioleaching microorganisms
Microbial consortia of acidophilic microorganisms from the BGR
strain collection were used to bioleach valuable metals from the tailings
samples. A mixed culture of mesophiles (consortium 1) composed of
Acidithiobacillus (At.) thiooxidans, At. ferrooxidans, Leptospirillum (L.)
ferrooxidans, and Ferroplasma (Fa.) acidiphilum and a mixed culture of
moderate thermophiles/thermophiles (consortium 2) composed of At.
caldus, L. ferriphilum, Sulfobacillus (Sb.) thermosulfidooxidans, Sb. bene­
faciens and Fa. acidiphilum were cultivated in basal salt medium
(Wakeman et al., 2008) on 50 mM ferrous iron plus 10 g/L S0 with an
initial pH of 2.0 at 30 ◦ C or 42 ◦ C, respectively.
2.3. Shake flask bioleaching tests with samples from four tailings sites
To evaluate the metal recovery from the four tailings sites by using
bioleaching vs. chemical leaching, experiments in shake flasks with 4 g
of sample material with 200 mL basal salt medium were set up. A 1 M
ferrous sulfate solution was added to get a final concentration of 50 ppm
Fe(II) in each flask which was inoculated with a mixed iron(II)- and
sulfur-oxidizing culture of the two acidophilic bacteria Acidithiobacillus
ferrooxidans and Acidithiobacillus thiooxidans. There were three parallel
bioleaching replicates for each sample and one control experiment
without inoculation of bacteria, but under the same chemical condi­
tions. The leaching was performed for 28 days.
2.4. Bioleaching of Taltal tailings
Bioleaching assays were performed at either 30 ◦ C or 42 ◦ C to
comparatively study the leaching efficiency of mesophilic and moderate
thermophilic consortia as well as to select the optimal bioleaching
conditions. Abiotic controls representing chemical leaching were per­
formed by adding 0.5 mM formic acid to the assays to inhibit potentially
introduced bioleaching bacteria without inoculation (Ballerstedt et al.,
2017). The leaching experiments were first conducted in shake flasks at
2% solids load and then at 5%, 10% or 15% solids load in stirred-tank
reactors (STR) using basal salts medium at pH 2.0. Elemental sulfur
(10 g/L) was supplemented as the additional energy sources. The pre-
cultivation strategy for increasing the pulp density was applied ac­
cording to the previous report (Hedrich et al., 2016). The experimental
conditions for tests in 2-L and 20-L STR (Supplementary Fig. S2) were as
follows: Temperature at 30 ◦ C or 42 ◦ C; leaching time of ~ 13 days (2-L
or 20-L STR); two or three inoculated parallel assays and one abiotic
control (one for 20-L STR due to the limited amount of material). Once
the experiments started, the pH in abiotic reactors was set according to
the pH values of the bioleaching reactors, which allows for a direct
comparison of bioleaching vs. chemical leaching at the same pH over the
experimental time. The pH adjustment was automated by the STR
operation systems through pumping 5 M sulfuric acid. Water evapora­
tion was compensated at regular intervals by addition of acidic deion­
ized water during the experiments, and samples were taken at different
time intervals. Bioleaching results are shown as mean of parallel STR
runs with standard deviation as error bars.
2.5. Analytical methods
Homogenously suspended samples were regularly taken from the
leaching assays for different analyses. Microscopic cell counting after
SYBR Green staining by using a fluorescence microscope, and quanti­
tative real-time PCR (qPCR) using copy numbers of bacterial 16S rRNA
genes were used to quantify the cell numbers, and terminal restriction
fragment length polymorphism (T-RFLP) was used to analyze the mi­
crobial community succession as previously described (Hedrich et al.,
2016). The pH and redox potential were analyzed with digital pH- /
redox meters. Consumed acid during leaching was recorded for com­
parison. Dissolved iron(II) and total iron were measured by the ferrozine
R. Zhang and A. Schippers Minerals Engineering 180 (2022) 107514

Table 1 3. Results and discussion

Shake flask copper bioleaching test with samples from four Chilean tailings sites.
Mean and standard deviation (in brackets) of three parallel bioleaching flasks as
well as one not inoculated control are given.
Tailings site Carola El Clarita
Salado
Cu content in tailings sample (mg/ 1155 3991 1234
kg)
Cu extraction in bioleaching assays 21 (3.1) 59 (1.5) 51 (3.8)
(%)
Cu extraction in control assays (%) 18 44 50
assay (Lovley and Phillips, 1987). ICP-OES was used to determine the
concentration of metal ions. XRF and XRD were used to analyze
elemental and mineralogical compositions of the Taltal tailings samples
as well as leaching residues. The “readily extractable” iron, copper and
zinc present in the tailings were determined by shaking 5 g tailings in
100 mL 0.1 M H2SO4 using a reciprocal shaker at 120 rpm at room
temperature for 1 h. The extracts were filtered through nitrocellulose
filters (0.2 μm pore size) and metal concentrations were measured by
ICP-OES. Microcalorimetric activity measurements were done according
to previous reports (Schippers and Bosecker, 2005). The measured heat
output was given in μW per gram tailings.
2.6. Confocal laser scanning microscopy (CLSM)
Cells on tailingś particle surfaces were visualized by using nucleic
acid stain Syto 9 or protein stain Sypro orange. Samples were neutral­
ized with filter-sterilized tap water and incubated with fluorescently
labelled stains for 20 min at room temperature. Afterwards, stained
samples were washed three times with filter-sterilized tap water in order
to remove the unbound stains. Samples were directly observed using
confocal laser scanning microscopy (CLSM) without any further treat­
ment. Examination of stained biofilms was performed by CLSM using a
TCS SP5X, controlled by the LASAF 2.4.1 build 6384 (Leica, Heidelberg,
Germany). The system was equipped with an upright microscope and a
super continuum light source (470–670 nm) as well as a 405 nm laser
diode. Images were collected with a 63 × water immersion lens with a
numerical aperture (NA) of 1.2 and a 63 × water-immersible lens with
an NA of 0.9.
Fluorescence image analysis and surface coverage calculation were
done by using an extended version of the software ImageJ (v1.50i).
Maximum intensity projection (MIP) was produced with the software
IMARIS version 8.1.2 (Bitplane AG, Zurich, Switzerland).
3.1. Preliminary bioleaching tests
Taltal Results of the copper bioleaching for the four tailings sites from Chile
are shown in Table 1. The tailings from the two sites Carola and Clarita
4582 showed a lower Cu bioleaching potential, while a higher copper content,
a higher copper extraction and larger differences between the bio­
69 leaching and the control assays were observed for the El Salado and
(0.9)
53
Taltal tailings. Since tailings from Taltal showed the highest Cu
extraction, this tailings site was chosen for more comprehensive bio­
leaching tests in bioreactors.
3.2. Taltal tailings characterization
The specific surface area of the coarse and fine Taltal tailings was 3.4
and 7.9 m2 per g, respectively. Results of elemental analyses of both
samples are shown in Table 2 and 3. Both tailings samples contained a
similar amount of organic carbon (0.03–0.04%), Co (0.011–0.012%)
and Cu (0.47–0.58%). The Cu content of about 0.5 % is of most eco­
nomic interest. The four mineral phases quartz, magnetite, plagioclase
and chlorite accounted for half of the components in both, coarse and
fine tailings. Sulfide minerals were not detected via XRD (Table 3) and
EPMA-SEM (Supplementary Fig. S3 and S4) but small amounts of about
1% pyrite and 0.2% chalcopyrite in the Taltal tailings were reported by
Drobe et al. (2021). Cu was also present in 0.4% atacamite (Cu2Cl(OH)3)
or chrysocolla (Drobe et al., 2021). Only approximately 0.5% of reduced
sulfur compounds were detected. To secure sufficient substrate for bio­
leaching microorganisms, elemental sulfur was added in tailings bio­
leaching experiments.
3.3. Bioleaching of Taltal tailings in 2-L STR
The bioleaching results of coarse tailings in 2-L STR at 30 ◦ C are
shown in Fig. 1. Total dissolved Fe concentrations in both, bioleaching
and abiotic assays, increased over time and reached 54 and 19 mg/L
after 13 days, respectively (Fig. 1a). The dissolved Fe2+ concentration
showed almost no changes in bioleaching assays and it remained around
1.5 mM until the end of the tests (Fig. 1b). The content of Fe2+ in the
abiotic assays continuously increased due to chemical leaching. Obvi­
ously, most dissolved Fe in bioleaching assays was Fe3+ due to
Fe2+–oxidizing activity of the bacteria. This agrees with the measured
values of redox potential (Fig. 1c), which was mainly determined by the
ratio Fe3+/Fe2+ in acidic conditions. The pH values in bioleaching assays
dropped continuously from 2.3 to 1 (Fig. 1d). The dissolved Cu and Co
concentrations in bioleaching assays showed a steady increase over
time, while only a slight increase of the Cu and Co concentrations was
detected in the abiotic assays (Fig. 2a and b). The metal extraction of Co,

Table 2
Elemental analysis of the two mixed bulk samples of Taltal tailings (%).
Tailings Corg Fe Co Cu TS CRS* As Pb Zn Ca

Coarse 0.03 29.6 0.012 0.47 0.94 0.49 0.006 0.004 0.012 2.93
Fine 0.04 19.6 0.011 0.58 0.93 0.48 0.009 0.004 0.018 3.39

Corg: organic carbon; TS: total sulfur content; CRS: chromium reducible sulfur (reduced sulfur e.g. pyrite).

Table 3
Quantitative mineralogical analysis of the two mixed bulk samples of the Taltal tailings by XRD with Rietveld refinement (%) of the coarse and fine tailings in Taltal,
Chile (from Drobe et al. 2021).
Tailings Mineral phase

Quartz Magnetite Gypsum Amphibole Calcite Hematite Muscovite Plagioclase Chlorite

Coarse 13 22 3 10 3 6 8 23 12
Fine 15 15 4 8 4 8 15 19 12

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R. Zhang and A. Schippers Minerals Engineering 180 (2022) 107514

Fig. 1. Bio- and abiotic leaching of coarse tailings with 10% solid loads in 2-L STR at 30 ◦ C by consortium 1. Changes of (a) total Fe concentration, (b) Fe2+
concentration, (c) redox potential (vs. Ag/AgCl) and (d) pH. ■ bioleaching assays, □ abiotic controls. The pH in the abiotic reactors was adjusted continuously as it
was in the bioleaching reactor runs before.

Fig. 2. Changes of (a) Cu and (b) Co concentrations during bioleaching of coarse Taltal tailings in 2-L STR at 30 ◦ C and (c) metal extraction after 13 days. ■
bioleaching assays, □ abiotic controls.

Fe, Cu was 38%, 8% and 86% for the bioleaching assays, and 20%, 2%
and 43% for the abiotic assays, respectively (Fig. 2c). These data indi­
cate that consortium 1 showed both, Fe2+- and /S0-oxidizing activity, as
expected, which enabled a better metal leaching from the tailings ma­
terial than just chemical leaching at the same pH as it was in the bio­
leaching reactor runs before.
Cell numbers and microbial community changes are shown in Fig. 3.
The consortium 1 showed a lag phase, but grew rapidly after day 4. Cell
numbers reached 9 × 108 cells/mL (Fig. 3a), which is comparable to the
cell density of the sample consortium grown on Rammelsberg tailings
containing pyrite (Zhang et al., 2020). The microcalorimetrically
measured heat output, due to S0 and Fe2+-oxidation activity, for the
coarse tailings was 33 μW/g on day 1. It increased continuously and
reached 84 μW/g on day 6. The heat output was 70 μW/g at the end of
the tests (Fig. 4). These values are in the range of those for bioleaching of
the pyritic Rammelsberg tailings (Zhang et al., 2020), but lower than for
stirred tank bioleaching of a copper concentrate (Hedrich et al., 2016).
Microcalorimetric measurements were also applied to monitor acid
mine drainage generation in mine waste dumps and the effect of coun­
termeasures (Schippers et al., 1995; Sand et al., 2007) as well as the
metabolic activity of the mineral-associated microbial community in
bioleaching (Makaula et al., 2017).

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R. Zhang and A. Schippers Minerals Engineering 180 (2022) 107514

Fig. 3. Changes of cell number and microbial community composition during the STR bioleaching experiment with coarse tailings using consortium 1 at 30 ◦ C. (a)
Cell number; (b) microbial succession, white: At. thiooxidans, black: At. ferrooxidans, vertical lines: L. ferrooxidans, horizontal lines: unidentified/indigenous bacteria.

Fig. 4. Microbial activity determined by microcalorimetric measurements during bioleaching of the coarse tailings material in STR at 30 ◦ C.

Community succession analysis showed dramatic changes of the
bioleaching community composition over time. At. ferrooxidans was the
dominant species in the initial stage, accounting for 50% and 42% of the
total community at day 1 and 6, respectively (Fig. 3b). At. thiooxidans
had a variable abundance with 21% on day, 3% on day 6 and 84% at the
end of the experiment becoming the dominant species. L. ferrooxidans
became the most dominant species on day 6 but the organismś abun­
dance decreased to 15% at the end of the tests likely due to the low pH. A
small proportion of unidentified/indigenous bacteria was detected on
day 13. The inoculated archaeal species Fa. acidiphilum seemed to be not
adapted to the leaching conditions and was not detected (not shown),
although it was frequently detected in bioleaching environments (Dop­
son et al., 2004; Van Hille et al., 2011). Previous work showed that both,
changing substrates and redox potentials, govern the microbial com­
munity changes during bioleaching processes (Rawlings and Johnson,
2007). In the current study, the supplementary S0 and continuously
formed Fe2+ due to tailings dissolution may simultaneously determine
the succession of the microbial community during bioleaching.
In order to test the influence of temperature and associated other
bioleaching community on metal dissolution of Taltal tailings,
bioleaching tests using consortium 2 at 42 ◦ C were conducted and results
are shown in Fig. 5. Total Fe and Fe2+ concentrations in bioleaching
assays using consortium 2 showed similar trends than for consortium 1
at 30 ◦ C (Fig. 5a and b; Fig. 1a and b). Total Fe concentration in abiotic
assays continuously increased and reached 62 mM, much higher than at
30 ◦ C. This indicates that Fe leaching from Taltal coarse tailings is
promoted by temperature. The redox potential in bioleaching assays
increased linearly in the first 7 days. It reached 674 mV (vs. Ag/AgCl)
after 12 days. In contrast, the redox potential increased slowly from 405
mV to 460 mV (Fig. 5c). Interestingly, Co showed a linear leaching
pattern, which indicates a suitable leaching environmental using con­
sortium 2 at 42 ◦ C. It also shows the potential of optimization of Co
leaching by extension of leaching time. In contrast, Co leaching in the
abiotic assays was rather poor (Fig. 5e). The final metal extraction of Co,
Fe, and Cu was 74%, 10% and 100% for the bioleaching assays, and
28%, 9% and 100% for the abiotic assays at 42 ◦ C, respectively (Fig. 5f).
The comparison with metal bioleaching at 30 ◦ C is also shown in this
figure. Bioleaching with fine tailings (instead of the coarse material)
using consortium 2 showed a similar metal leaching performance
(Supplementary Fig. S5), with a final metal leaching extraction of Co, Fe,

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R. Zhang and A. Schippers Minerals Engineering 180 (2022) 107514

Fig. 5. Bio- and abiotic leaching of coarse tailings with 10% solid loads in 2-L STR at 42 ◦ C by consortium 2. Changes of (a) total Fe concentration, (b) Fe2+
concentration, (c) redox potential (vs. Ag/AgCl), (d) pH and (e) Co concentration over time. (f) Comparison of metal leaching by consortium 1 and consortium 2 with
abiotic control. ■ bioleaching assays, □ abiotic controls.

Fig. 6. Microbial community composition and total cell numbers during the STR bioleaching experiment at 42 ◦ C with consortium 2. (a) Cell number, (b) community
analysis, white: At. caldus, black: L. ferriphilum, vertical lines: Sb. benefaciens/Sb. thermosulfidooxians.

6
R. Zhang and A. Schippers
Fig. 7. Bioleaching of the Taltal fine tailings in 20-L STR with 15% solid loads at 42 ◦ C by consortium 2 for 12 days. Changes of (a) Fe2+ concentration and total iron
concentration, (b) cell number, (c) redox potential, (d) pH, (e) dissolved Co and (f) Cu concentrations.
and Cu of 68%, 14% and 100% for the bioleaching assays, and 32%, 12%
and 100% for the abiotic assays at 42 ◦ C, respectively. The acid con­
sumption (calculated for concentrated sulfuric acid per kg dry tailings)
was 60 g/kg and 20 g/kg for abiotic leaching and bioleaching of coarse
tailings, respectively, and 113 g/kg and 29 g/kg for abiotic leaching and
bioleaching of fine tailings, respectively.
Considering the experiments one can conclude that Co was mainly
bioleached, probably associated to the about 1% pyrite in the samples
(Drobe et al., 2021), while Cu was mainly acid leached, which was
present in 0.4% atacamite (Cu2Cl(OH)3) and chrysocolla, besides chal­
copyrite (Drobe et al., 2021). Overall these results show that 1) an
increased temperature promotes Fe and Cu leaching, 2) Co leaching was
promoted mainly due to microbial activity (Fe2+ - and sulfur-oxidation),
and 3) bioleaching using moderate thermophiles showed a much higher
Co performance than using mesophiles.
Consortium 2 showed good growth in 2-L STR at 42 ◦ C and the
community succession analysis showed that the sulfur-oxidizer At. cal­
dus dominated the bioleaching process (at the beginning and at the end;
Fig. 6b). This was expected since the supplemental S0 was the main
substrate. Cells of Sb. benefaciens/Sb. thermosulfidooxians accounted for
4% on day 1 and its proportion increased to 16% at the end of the tests.
Sb. thermosulfidooxidans was shown to have a weak Fe2+ oxidation ac­
tivity (Christel et al., 2018), in addition to its sulfur oxidation capacity
(Golovacheva and Karavaiko, 1978). Sulfobacillus species belong to the
7
Minerals Engineering 180 (2022) 107514
indigenous bacteria in mine tailings (Zhang et al., 2021).
3.4. Bioleaching of Taltal tailings in 20-L STR
For a scale up test, the tailings adapted culture of consortium 2 was
used to bioleach fine Taltal tailings in 20-L STR at 15% solid load
without running an abiotic control due to material limitation. The re­
sults are shown in Fig. 7. Total Fe increased continuously during the
whole period, while the Fe2+ concentration was kept below 1 mM due to
the high Fe2+ oxidizing activity of consortium 2 (Fig. 7a). The cell
number increased over time and reached 1.5 × 109 cells/mL after 12
days (Fig. 7b). Redox potential, in accordance with the changes of total
Fe and Fe2+ concentrations, continuously increased and reached 670 mV
(vs. Ag/AgCl). This value is usually sufficient to dissolve pyrite in bio­
leaching assays (Liu et al., 2008). The pH in the 20-L STR assay dropped
below 0.9, which proves the activity of the sulfur-oxidizing microor­
ganisms in consortium 2. The leaching of Co increased rapidly within the
first 7 days and seemed to reach a plateau afterwards (Fig. 7e). The Co,
Fe and Cu extraction reached 49%, 11% and 83% at the end of the 20-L
STR experiment (Supplementary Fig. S6). Compared with the somehow
lower Co and Cu extraction than in the assays with 2-L STR (Supple­
mentary Fig. S5), further optimization of scaling-up tests, e.g. stir con­
trol or agitation, or using continuous stirred-tank reactors, as opposed to
batch leaching applied here, has to be considered for improving Co and
R. Zhang and A. Schippers
Cu recovery from tailings. In general, the low Fe extraction of below
14% in all assays would favor the downstream processing for Cu and Co
recovery from the pregnant leach solution (PLS).
It is worth to mention that Taltal tailings bioleaching was probably
rather conducted by planktonic cells than by cells attached to tailings
particles, since attached cells were quite limited as indicated by the
CLSM analysis (Supplementary Fig. S7). One reason might be that the
added elemental sulfur as the main substrate was dissolved during the
bioleaching process. However, bioleaching is performed by both,
planktonic cells and attached cells (Vera et al., 2013; Schippers et al.,
2014).
4. Conclusions
Bioleaching seems to be a suitable processing option for cobalt and
copper recovery from two of four mine tailings sites in Chile. Tailings
samples from the selected Taltal site were bioleached by using meso­
philic as well as moderate thermophilic consortia of bioleaching or­
ganisms. Moderate thermophiles at 42 ◦ C showed a higher bioleaching
performance compared to mesophilic acidophiles at 30 ◦ C in 2-L STR,
namely 74% vs. 38% for Co and 100% vs. 86% for Cu. Co was mainly
bioleached, probably associated to the pyrite in the samples, while Cu
was mainly acid leached. In the mini-pilot scale (20 L) bioreactor, Co
and Cu bioleaching reached 49% and 83%, respectively. Metal re­
coveries may be improved using continuous stirred-tank reactors, as
opposed to batch leaching applied here. A cost model for reprocessing of
the Taltal mine tailings including chemical copper leaching data is
presented by Drobe et al. (2021) indicating overall economic feasibility
of reprocessing. Chemical copper leaching has also been demonstrated
in our study, however the costly sulfuric acid is replaced by cheaper
elemental sulfur being oxidized to sulfuric acid by the acidophilic mi­
croorganisms. Additional value might be obtained for cobalt recovered
via bioleaching. This study points out the suitability of bioleaching for
re-processing of mine tailings.
CRediT authorship contribution statement
Ruiyong Zhang: Conceptualization, Investigation, Data curation.
Axel Schippers: Conceptualization, Investigation, Data curation.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
We thank Dr. Thomas Neu and Ute Kuhlicke (Department of River
Ecology, Helmholtz Centre for Environmental Research GmbH – UFZ,
Magdeburg, Germany) for CLSM analysis. We thank several colleagues
in BGR for chemical and mineralogical analyses, and especially Wiebke
Schulze for excellent technical assistance, Malte Drobe and Sabrina
Hedrich for providing samples, methodological support and scientific
discussions, Kristian Ufer for XRD analysis, and Jens Gröger-Trampe for
CRS analysis. This work was financially supported by the German
Mineral Resources Agency (DERA) within BGR.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.mineng.2022.107514.
8
Minerals Engineering 180 (2022) 107514
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