Minerals Engineering 15 (2002) 689–694

This article is also available online at:

www.elsevier.com/locate/mineng

Bioleaching capacity of an extremely thermophilic culture

for chalcopyritic materials
A. Rubio a, F.J. Garc
ıa Frutos b,*

a
Instituto Geol

ogico y Minero de Espa~
na (IGME), La Calera n° 1, 28760 Tres Cantos, Madrid, Spain
b
Centro de Investigaciones Energ

eticas, Medioambientales y Tecnol

ogicas (CIEMAT), Av. Complutense 22, Edificio 20, 28040 Madrid, Spain
Received 9 March 2001; accepted 2 May 2002

Abstract
A thermophilic culture specific to bioleaching chalcopyritic materials has been obtained from a typical chalcopyritic copper
concentrate of the Spanish Pyritic Belt. This paper shows the effect of pulp density (w/v) on bioleaching culture capacity with respect
to this copper concentrate. The results of the batch tests show that it is possible, operating at 10% of pulp density to attain copper
extraction of 94% in 10 days and, at higher pulp densities (20%), to attain good copper extraction (80%) in only 14 days. In the same
way, the culture has been amply tested with different chalcopyritic ores and copper concentrates. The results obtained with four of
these materials, two refractory gold ores and two copper concentrates are also presented. These results show a varying and versatile
bioleaching capacity of this bacterial culture. A model of bacterial attack of this culture to leach chalcopyrite is postulated.
Ó 2002 Elsevier Science Ltd. All rights reserved.

Keywords: Mineral processing; Sulphide ores; Hydrometallurgy; Bioleaching and bacteria

1. Introduction metal concentrates, this technology remains as a prom-

The resources of high ores in the world are becoming
more and more scarce making the processing of more
complex ores necessary. Conventional mineral process-
ing on complex sulphide ores carried out by differential
flotation often produces high grade concentrates but
with contaminants that penalise and make its marketing
and pyrometallurgical processing difficult. Therefore, a
lot of effort has been made in the development of hy-
drometallurgical process suitable for the treatment of
these ores, but most of the proposed methods are
complex and expensive, Alvarez (1996).
Among these hydrometallurgical processes, biohy-
drometallurgical techniques appear to be an alternative
for the treatment of these concentrates. These methods,
that were first applied industrially to copper and ura-
nium productions using bioassisted heap, dump and
in situ technologies, are successfully used today in ex-
traction of gold from refractory sulphide-bearing ores
and concentrates, Jordan et al. (1996). However, for other
*
Corresponding author. Tel.: +34-91-3466232; Fax: +34-91-
3466269.
E-mail address: garcia.frutos@ciemat.es (F.J. Garc
ıa Frutos).
0892-6875/02/$ - see front matter Ó 2002 Elsevier Science Ltd. All rights reserved.
PII: S 0 8 9 2 - 6 8 7 5 ( 0 2 ) 0 0 1 2 4 - 3
ising alternative against conventional pyrometallurgical
extraction processes. This is the case of the treatment
of chalcopyritic concentrates, which represent a more
complicated situation, due to the natural refractivity of
chalcopyrite.
One of the mainly problems of the bioleaching pro-
cesses applied to copper concentrates and refractory ores
are the low kinetics of the reactions, with high residence
time that does not permit it to be an economic process.
For this reason, current researches into this field are
focused on how to increase this bioleaching rate. Apart
from improving engineering design of bioreactors, the
possibilities to increase bioleaching rates depend on the
use of catalyst and isolation and adaptation of new mi-
croorganisms with high capacity to leach these ores.
Several studies with mesophilic microorganisms as
Thiobacillus ferrooxidans and Leptospirillum ferrooxi-
dans had showed very slow copper leaching rates, Mehta
(1982) and Sand et al. (1992). However, when thermo-
philic microorganisms were used leaching rates were
considerably increased due to high temperatures, higher
metal tolerance capacity and metabolic characteristics of
this type of microorganisms, Brierley (1993) and Clark
and Norris (1996).
690 A. Rubio, F.J. Garc
ıa Frutos / Minerals Engineering 15 (2002) 689–694
Many researches have investigated the possibility of
using thermophilic microorganisms to improve metal
leaching rates instead to mesophilic microorganisms.
Future process developments must and will include
thermophilic bacteria, which will play an increasingly
important role in biooxidations of minerals. Thermo-
philic Archaea, Sulfolobus species, Acidianus brierleyi
and Metallosphaera sedula which grow from 60 to 75 °C,
are particularly adept in bioleaching of copper from the
highly refractory chalcopyrite, Brierley and Brierley
(1999) and Dew et al. (1999). Moreover, the use of these
microorganisms that naturally thrive on ore samples and
its aqueous environments, could be a good option as
they probably have specificity for leaching chalcopyrite
and a much higher capacity for adaptation. In this sense,
there is a work carried out by Lopez-Archilla et al.
(2001), on the microbial ecology of Rio Tinto waters,
where they report to have isolated a thermophilic ar-
chaeabacteria, Sulfolobus rivotincti, with specificity to
bioleach chalcopyrite and therefore with possibilities to
be used in chalcopyritic concentrates treatment, Bal-
lester (1996) and G
omez et al. (1999).
The use of these microorganisms for bioleaching
chalcopyrite implies the knowledge of the mechanisms
involved in the process.
Initially, chalcopyrite can be oxidised by dissolved
oxygen according to Eq. (1), Hiroyosi et al. (1997), fa-
voured for acidic solution, and as it is known, two
modes of bacterial attack can be distinguished:
In indirect attack (Eq. (2)), the role of bacteria is to
regenerate the oxidant ferric ion in the bulk phase, from
the ferrous iron resulting from the chemical oxidation
of the metal sulphide in the ore by ferric iron (Eq. (3)).
In the direct attack, the bacteria leach the metal sul-
phide by attaching to the mineral surface and oxidising
it enzimatically by conveying electrons from the reduced
moiety of the mineral, Erlich (1999).
CuFeS2 þ O2 þ 4Hþ ¼ Cu2þ þ Fe2þ þ 2S0 þ 2H2 O
CuFeS2 þ 4Fe 3þ
¼ Cu 2þ
þ 5Fe 2þ
þ 2S 0
2þ þ 3þ
4Fe þ 4H þ O2 ¼ 4Fe þ 2H2 O
Sand (1997) have suggested that because Fe3þ oxi-
dises metal sulphide with both the direct and indirect
mechanisms, there is no difference between the two
mechanisms. Their model emphasises a similarity in the
chemistry of attack of the sulphide moiety by iron, and
makes no distinction between ferric iron in the bulk
phase and ferric iron bound in the cell envelope.
The authors have obtained a mixed natural thermo-
philic culture from a typical chalcopyiritic copper con-
centrate of the Spanish Pyritic Belt, Rubio (1998). This
culture has selectivity with respect to leaching chal-
copyrite, when this is present together with other sulp-
hides in ores and concentrates.
This paper describes the bioleaching capacity of the
culture to operate at high pulp density and its versatility
for leaching chalcopyritic materials, from ores to con-
centrates, postulating an ‘‘in situ’’ indirect bacterial at-
tack.
2. Methods
2.1. Microbial inoculum
A mixed thermophilic culture (MTC) of native mi-
croorganisms, isolated on a typical chalcopyritic con-
centrate described below (Concentrate 0), was used as
inoculum in the initial bioleaching experiments.
The methodology of the isolation was the following:
The concentrate was strong stirring (360 rpm) at pH
higher than 4 and 65 °C of temperature during 2 h for
remove the microorganisms attached on the chalcopy-
rite surface. Once the microorganisms were unattached
and free in solution, the solid was removed of the liquid
by filtration. The bacteria-laden liquid obtained was
filtered through a 0.22 lm Millipore membrane filter to
collect the microorganisms, which were suspended in
1:10 9 K medium at pH 1.30.
Preliminary comparative bioleaching tests were per-
formed using a bacterial inoculum of Sulfolobus sp.
kindly supplied by Dr. F. Carranza, Universidad de
Sevilla, Spain.
Both thermophilic cultures were maintained on cop-
per concentrate (Concentrate 0) at 1% w/v pulp density
medium with 1:10 salts of 9 K medium at 65 °C of
temperature and initial pH of 1.30.
The cultures were successfully adapted to higher pulp
density of copper concentrate until obtaining high cop-
per extractions in short residence times. In these tests,
the inoculum was obtained as the culture isolation was
made.
The final pulp of the bioleching test was filtered,
ð1Þ obtaining the leach liquor and the solid residue. The
ð2Þ solid was intensively stirring during 2 h with water at pH
4. The pulp obtained was filtered again and the final
ð3Þ liquid obtained, which contains most the bacteria, which
were attached to the solid residue, was filtered through a
0.22 lm Millipore filter where bacteria were retained.
Finally, the bacteria were re-suspended in 50 ml of the
leach liquor obtained in the first filtration in order to get
the inoculum volume (5% v/v) for the next bioleaching
test.
2.2. Ore samples
A chalcopyritic copper concentrate obtained from
conventional differential flotation was used to develop
the mixed thermophilic culture. This concentrate (Con-
centrate 0) came from the Spanish Pyritic Belt and its
 A. Rubio, F.J. Garc
ıa Frutos / Minerals Engineering 15 (2002) 689–694 691

Table 1 ished, solids were removed by filtration, and chemically

Chemical composition of copper concentrate characterised as well as the leachate.
Sample Chemical analysis (wt%) Sterile bioleaching tests were carried out with ore
Cu Zn Pb Fe S sterilised by autoclaving at 121 °C, 30 min and 1atm of
Concentrate 0 23.37 2.58 2.52 32.63 38.00 pressure, and adding a solution of 10% ethanol to the
leaching media.

composition is given in Table 1. The mineralogical 2.4. Analysis

composition of this sample shows chalcopyrite and py-
rite as main mineralogical species and galena and
sphalerite as secondary mineralogical phases. The par-
ticle size distribution presents a passing d80 of 20 lm.
Concentrate 1, also used in bioleaching tests is es-
sentially the same as Concentrate 0, corresponding to
another lot of copper concentrate obtained in the same
flotation point at the industrial plant. This sample pre-
sents less copper content and higher zinc content than
Concentrate 0.
The other chalcopyritic samples, refractory gold ores
(Ores 1 and 2) and copper concentrates (Concentrate 1
and 2), also came from the Spanish Pyritic Belt, were
selected in this work to study the selectivity of the MTC
to chalcopyrite. The chemical composition of these
chalcopyritic samples is given in Table 2. The copper
present in these samples is as chalcopyrite.
2.3. Bioleaching experiments
Soluble species of copper, zinc, lead, total iron and
minor elements were analysed by ICP, Del Barrio
(1992), using a spectrophotometer ICAP-61 Thermo
Jarrel Ash. The ferrous iron was analysed by a volu-
metric method by titration with potassium dichromate,
Kolthoff (1979). Copper, zinc, lead and iron content in
the ore samples and leaching solid residues were analy-
sed by XRF, Mart
ın Rub
ı (1998), using a spectropho-
tometer Philips PW-1404, and minor elements by ICP.
Total sulphur was gravimetrically determined and ele-
mental sulphur was analysed by toluene extraction in
a Soxhlet apparatus.
The pH was measured with a 704 pH-meter Metr-
ohm. The redox potential (Eh) was measured with a
platinum electrode with an Ag/AgCl reference electrode.
Mineralogical composition was determined by XRD
using a difractometer PW-1700 Philips.

Bioleaching experiments were carried out in 1 l glass 3. Results and discussion

cylindrical reactors provided with a cap with four holes
to allow mechanical stirring (at 130 rpm), aeration (10– 3.1. Influence of pulp density
15 l/h) and sampling. These reactors were placed in a
thermostatic bath to keep the temperature constant at A study of bioleaching capacity on copper concen-
65 °C. trates by MTC culture was done at different pulp den-
During the experiments the pH was kept at 1.30 by sities such as 1%, 5%, 10%, 15% and 20% (w/v).
the addition of 10N H2 SO4 when were necessary. This Preliminary tests carried out at 1% pulp density (w/v)
10 N H2 SO4 addition was made to avoid the precipita- show a higher capacity of MTC culture to bioleach the
tion of iron in form of jarosites, which damage the bio- copper present in the copper concentrate (Concentrate
leaching process. 0) compared to the tests carried out with Sulfolobus sp.
Redox potential and pH were measured daily, while culture (Sb) on the same copper concentrate. Both cul-
the levels of copper, zinc, and iron in solution were tures had been maintained and adapted to this copper
analysed daily or every 2 days, depending on the test. concentrate by successive sub-culturing for the realisa-
Water was added to the reactors in order to compensate tion of these bioleching tests.
for evaporation losses. Once bioleaching tests were fin- Fig. 1 shows copper extractions obtained on copper
concentrate with MTC and Sb cultures in representative
adaptation tests at 1% of pulp density (w/v). In the
Table 2 sterile test, only 11 % copper extraction was obtained in
Chemical composition of chalcopyritic samples more than 500 h. In bioleaching tests with Sulfolobus sp.
Sample Chemical analysis (wt%) culture (Sb), initially a cooper extraction up 98% was
Cu Zn Pb Fe S reached in 500 h, but after successive adaptation tests,
Ore 1 0.69 1.64 0.71 39.55 33.35 the kinetic of reaction was increasing, obtaining maxi-
Ore 2 1.47 1.1 0.53 42.15 45.6 mum copper extractions (>98%) in 350 h. However,
Concentrate 1 20.76 4.65 2.87 30.11 37.29 with MTC culture the same copper extraction (more
Concentrate 2 6.70 <0.10 <0.10 24.20 n.a. than 98%) was obtained in only 165 h with only two
n.a: no appear. successive adaptive tests.
692 A. Rubio, F.J. Garc
ıa Frutos / Minerals Engineering 15 (2002) 689–694
Fig. 1. Copper extraction in bioleaching adaptive tests at 1% (w/v)
pulp density. (Sterile test, Sulfolobus sp. and MTC tests at pH 1.30 and
65 °C). Sample: Concentrate 0. (The sub-indexes 1; 2; 3; . . . ; n corre-
spond to consecutive tests.)
Bioleaching tests carried out at 5% pulp density (w/v)
are shown in Fig. 2. Similar ability to preferentially
leach copper by MTC thermophilic culture was ob-
tained. The extractions achieved with the MTC were
much more significant than Sb culture. It can be ob-
served that in the first test on Concentrate 0 with the
culture isolated MTC, a copper extraction of about 90%
was obtained in only 21 days. In second test with this
culture, in 17 days the copper extraction reached 97%,
considerably higher than the extraction attained (80%)
after 56 days of bioleaching when Sulfolobus sp. culture
was used.
From these results, a study in order to define the
optimum conditions of this bioleaching process was
carried out increasing pulp density (1%, 5%, 10%, 15%
and 20% (w/v)). This study is described elsewhere,
Rubio (1998). The best conditions obtained from 1%
to 20% pulp densities vary only in oxygen uptakes.
In Fig. 3 appears the evolution of copper extractions
to all tests done at pulp densities studied in its optimum
conditions. As it can be observed the results of batch
tests show that it is possible to work both with high pulp
Fig. 2. Copper extraction in bioleaching adaptive tests at 5% (w/v)
pulp density. (Sulfolobus sp. (Sb) and MTC tests at pH 1.30 and 65
°C). Sample: Concentrate 0. (The sub-indexes 1; 2; 3; . . . ; n correspond
to consecutive tests.)
Fig. 3. Comparative copper extraction at the different pulp densities
studied (1, 5, 10, 15 and 20% (w/v) at pH 1.30 and 65 °C). Sample:
Concentrate 0.
density (20%) obtaining good copper extraction (80%)
and with lower pulp density (10%) obtaining better
copper extraction (94%) in 14 and 10 days respectively.
In all tests done at different pulp densities, iron in
solution at first stages was mainly as ferrous iron and
only the presence of ferric iron was more notable when
almost 80% of copper is leached.
Fig. 4 shows total iron and ferrous iron during a bio-
leaching test on Concentrate 0 at 10% of pulp density
with MTC culture. As it can be observed, mainly form
of iron in solution was as ferrous iron. This ferrous iron
in solution decreases at the final stages (from the eighth-
day on) which corresponds to an increase in the solution
redox potential over 450 mV and a copper dissolution of
80%.
Redox potential evolution is showed in Fig. 5. As can
be seen, Eh in first stages of bioleaching is about 400 mV
to reach 425 mV in an intermediate step, confirming the
most presence of ferrous iron in solution. The more
notable redox potential increase was produced from day
7, when ferrous iron starts to decrease in solution
reaching a final Eh of 475 mV.
Fig. 6 shows zinc extraction in the same bioleaching
test. A fast initial leaching of zinc can be appreciated. At
Fig. 4. Concentration of iron in leach solution to a representative test
at 10% (w/v) pulp density, at pH 1.30 and 65 °C. Sample: Concentrate 0.
 A. Rubio, F.J. Garc
ıa Frutos / Minerals Engineering 15 (2002) 689–694 693

Fig. 7. Copper extraction in bioleaching tests with copper Concentrate

Fig. 5. Redox potential (Eh) evolution to a representative test at 10% 1 and 2 (10% (w/v) pulp density, pH 1.3 and 65 °C).
(w/v) pulp density, at pH 1.30 and 65 °C. Sample: Concentrate 0.

done at 10% pulp density. The copper present in these

ores was quickly leached in 3 and 4 days, and the zinc
was only extracted when copper is almost leached and
the redox potential was over 475 mV.
Fig. 7 shows the copper extraction obtained in bio-
leaching tests carried out at 10% pulp density with the
other copper concentrates used (Concentrate 1 and 2).
Concentrate 2 with less initial copper content than
Concentrate 1, was more easily bioleached by MTC
culture, obtaining more than 90% of copper extraction
in 6 days. With Concentrate 1 (similar to Concentrate 0)
and without a period of adaptation, more than 70% of
copper extraction was obtained in 12 days. In the sterile
Fig. 6. Concentration of zinc in leach solution to a representative tests, copper extraction obtained was 14% for Concen-
test at 10% (w/v) pulp density, at pH 1.30 and 65 °C. Sample: Con- trate 1 and 21% for Concentrate 2.
centrate 0. From these tests it can be observed that the leaching
behaviour of this culture with respect to chalcopyrite
intermediate stage of leaching, as there are not sufficient present in these samples, was similar to the initial copper
ferric iron in solution, the zinc dissolution is low, and concentrate, leaching in a selective way this mineralog-
only an increase in zinc extraction was observed when ical species.
copper was bioleached (after 8 days of test) coinciding
with the increase of ferric iron in solution. 3.3. Mechanism of bioleaching
The zinc present was almost completely leached only
when the duration of tests was prolonged longer after all Three significant facts were observed in all bioleach-
copper had been leached. ing tests carried out with the MTC culture:
X-ray diffraction of solid residues showed elemental
sulphur and pyrite as main species present, and anglesite 1. Iron in solution was mainly found as ferrous iron.
and sphalerite as minor species. 2. Only when copper extraction was significant the pres-
ence of ferric iron in solution was more notable.
3.2. Bioleaching of chalcopyritic samples 3. Zinc extractions were only about 60%, being almost
constant along the leaching time.
In Table 3, appear in short, the results obtained with
the two refractory gold ores selected. The tests were In addition to the high temperature of leaching, these
characteristics may have a significant incidence on the
high bioleaching rate obtained with MTC culture. From
Table 3
Results obtained in bioleaching tests with the refractory gold ores
these facts the following ‘‘in situ’’ indirect leaching
model may be considered:
Sample Pulp density Time Yield (%)
(% w/v) (days)
In acidic solution, chalcopyrite is oxidised by dis-
Cu Zn Fe
solved oxygen according to the reaction in Eq. (1), Hi-
Ore 1 10 3 90.20 70.59 11.52 royosi et al. (1997). In addition to this reaction, the Eq.
Ore 2 10 4 89.16 59.02 23.99
(3) occurs in the presence of ferrous ions. If ferric ions
694 A. Rubio, F.J. Garc
ıa Frutos / Minerals Engineering 15 (2002) 689–694
were produced by the reaction in Eq. (3), chalcopyrite
oxidation by ferric ions would be taking place in addi-
tion to reaction in Eq. (1) through the reaction in
Eq. (2).
Considering the final products obtained in this case,
an indirect leaching mechanism could be postulated.
However, ferric iron in solution was low and MTC
culture have very low capacity to oxidise ferrous iron in
solution, Rubio (1998). Besides, according to rest po-
tentials, sphalerite should be easier to be leached but this
is not true, in this case. Because of that, the authors
believe that probably, ferrous ions are adsorbed first
onto chalcopyrite surface, being the culture able to
catalyse ferrous oxidation to ferric iron at the surface.
Then, ferric ions produced on the surface oxidise chal-
copyrite according to reaction of Eq. (2) being simulta-
neously reduced to ferrous ions. When the chalcopyrite
leaching is almost completed, ferric ion concentration in
solution is already high (10 g/l) and then the Fe3þ is able
to leach sphalerite by ferric leaching through reaction
in Eq. (4).
ZnS þ 2Fe3þ ¼ Zn2þ þ 2Fe2þ þ S0 ð4Þ
This model is in accordance with the results obtained
by the authors in a study carried out improving the rate
of bioleaching of MTC culture on this copper concen-
trate when initial additions of ferrous ions were made.
When initial additions of 1.8 g/l of ferrous iron were
done, the rate increased from 60 mg/lh to 96 mg/lh,
Rubio (1998).
4. Conclusions
A natural MTC was obtained from a chalcopyritic
copper concentrate with an ability to preferentially leach
chalcopyrite in concentrates against other mineralogical
species less refractory as sphalerite. After a period of
adaptation of the mixed thermophilic culture to in-
creasing pulp densities (1, 5, 10, 15 and 20% w/v), high
copper extractions (>94%) and high leaching rates (2.3
g/l per day at 10% of pulp density) can be obtained.
From the results observed in all bioleaching tests, it
was proposed an ‘‘in situ’’ indirect leaching mechanism
of chalcopyrite.
From these laboratory results this culture can be
considered as a promising advance in the biohydro-
metallurgical treatment of chalcopyritic concentrates
and its potential use on an industrial scale.
Acknowledgements
We would like to thank A. Ilarri, researchers and
technicians of IGME Laboratory where this study was
carried out for your assistance in the analyses and ex-
periments.
References
Alvarez, J.L., 1996. Revisi
on cr
ıtica de los procesos de beneficio de
sulfuros complejos. Simposio sulfuros polimet
alicos de la Faja
Pir
ıtica Ib
erica, Huelva.
Ballester, A., 1996. Biohidrometalurgia y sulfuros complejos. Investi-
gaciones actuales. Simposio sulfuros polimet
alicos de la Faja
Pir
ıtica Ib
erica, Huelva, 1996.
Brierley, C.L., 1993. Practical role of thermophilic bacteria in
bioleaching and biooxidation. In: International Conference and
Workshop Application of Biotechnology to the Mineral Industry,
Australian Mineral Foundation, pp. 2.1–2.7.
Brierley, J.A., Brierley, C.L., 1999. Present and future commercial
applications of biohydrometallurgy. In: Amils, R., Ballester, A.
(Eds.), International Biohydrometallurgy Symposium IBSÕ99. Part
A. Elsevier, pp. 81–89.
Clark, D.A., Norris, P.R., 1996. Oxidation of minerals sulfides by thermo-
philic microorganisms. Minerals Engineering 9 (11), 1119–1125.
Del Barrio, S., 1992. Estudio e investigaci
on de un espectrofot
ometro
de emisi
on plasma ICP aplicado a la prospecci
on geoqu
ımica
multielemental. Bolet
ın Geologico y Minero (Publicaci
on especial).
Dew van Buuren, McEwan, Bowker, 1999. Bioleaching of base metal
sulphide concentrates: a comparison of mesophile and thermophile
bacterial cultures. In: Amils, R., Ballester, A. (Eds.), International
Biohydrometallurgy Symposium IBSÕ99. Part A. Elsevier, pp. 229–
238.
Erlich, H., 1999. Past, present and future of biohydrometallurgy. In:
Amils, R., Ballester, A. (Eds.), International Biohydrometallurgy
Symposium IBSÕ99. Part A. Elsevier, pp. 3–12.
G
omez, E., Ballester, A., Gonz
alez, F., Bl
azquez, M.L., 1999.
Leaching capacity of a new extremely thermophilic microorganism.
Sulfolobus rivotincti. Hydrometallurgy 52, 349–366.
Hiroyosi, N., Hirota, M., Hirajima, T., Ysunekawa, M., 1997. A case
of ferrous sulfate addition enhancing chalcopyrite leaching.
Hydrometallurgy 47, 37–45.
Jordan, M.A., McGuiness, S., Philips, C.V., 1996. Acidophilic
bacteria-their potential mining and environmental applications.
Minerals Engineering 9 (2), 169–181.
Kolthoff, I., 1979. M. An
alisis qu
ımico cuantitativo, Ed.Niger SRL.
Lopez-Archilla, A.I., Mar
ın, I., Amils, R., 2001. Microbial community
composition and ecology of an acidic aquatic environment: the
Tinto river, Spain. Microbial Ecology 41, 20–35.
Mart
ın Rub
ı, J.A., 1998. An
alisis por fluorescencia de Rayos X de los
sulfuros masivos en perlas de fusi
on. In: IV Simposio Internacional
de Sulfuretos Polimet
alicos de la Faixa Piritosa Ib
erica, C10, pp. 1–13.
Mehta, A.P., Murr, L.E., 1982. Kinetics study of sulfide leaching by
galvanic interactions between chalcopyrite, pyrite and sphalerite
in the presence of T. ferrooxidans (30 °C) and a thermophilic
microorganism (55 °C). Biotechnology and Bioengineering XXIV,
919–940.
Rubio, A., 1998. Investigaci
on en la mejora de la cin
etica de
biolixiviaci
on de concentrados calcopir
ıticos. Utilizaci
on de micro-
organismos term
ofilos, PhD Thesis, Dpto. Biolog
ıa Molecular,
Universidad Aut
onoma de Madrid.
Sand, W., Rhode, K., Sobotke, B., Zenneck, C., 1992. Evaluation of
leptospirillum ferrooxidans for leaching. Applied and Environ-
mental Microbiology 58, 85–92.
Sand, W., 1997. Novel mechanism for bioleaching of metal sulfides. In:
Biotechnology Comes of Age. International Biohydrometallurgy
Symposium IBS97 BIOMINE97. Australian Mineral Foundation,
Glenside, SA.