Response of microbial systems to thermal stress in

biooxidation-heap pretreatment of refractory gold ores

James A. Brierley
*
Brierley Consultancy LLC, P.O. Box 260012, Highlands Ranch, CO 80163-0012, USA
Abstract
Operation of a commercial-scale biooxidation heap for pretreatment of refractory gold ores demonstrates the propensity of
heaps to heat concurrent with pyrite oxidation. Heap temperatures can reach 75 jC. The high temperature is lethal for
mesophilic iron-oxidizing bacteria. Column testing was conducted to compare effects on biooxidation pretreatment of a sulfidic
refractory gold ore and the microbes response to temperature when bacteria and archaea were grown at different temperatures.
Five columns were operated at a different temperature regime: 2023 jC; 35 jC; 50 jC; 60 jC; and one column varied from
2023 to 60 jC. For the variable temperature column, the temperature was increased stepwise from 2023 jC, to 35 jC, to 50
jC, to 60 jC in 2-week increments; after 2 weeks at 60 jC, the temperature was decreased every 2 weeks in the same stepwise
manner. The following inocula were used: (1) columns at 2023 and 35 jC, mixed culture of Acidithiobacillus ferrooxidans
and Leptospirillum ferrooxidans; (2) column at 50 jC, moderate-thermophilic iron-oxidizing Sulfobacillus-type bacteria; (3)
column at 60 jC, hyper-thermophilic archaea, Acidianus and Metallospheara species; and (4) an equal mix of all of the above
for the variable temperature column. The hyper-thermophiles did not increase in numbers until the temperature was increased to
50 jC and above. Lowering the temperature resulted in a decrease in population density of this group of microbes. The
moderately thermophilic bacteria increased in number until the temperature was raised to 60 jC; at that point, numbers
decreased and remained stable as further decrease in temperature occurred. Decimation of the mesophilic iron-oxidizing bacteria
occurred when the temperature was increased to 50 and 60 jC. A low-level population concentration was detectable at
increased temperatures attributable to their presence in the ambient temperature solution reservoir. However, this population
increased as the temperature was lowered. There was little difference in the amount of sulfide oxidation at either ambient room
temperature or 35 jC. Increasing the biooxidation temperature from 35 to 50 jC increased apparent sulfide oxidation from
about 38% to 48%. At 60 jC, sulfide oxidation was highest at about 51%. The variable temperature column also had higher
sulfide oxidation, 41%, attributable to periods of high temperature biooxidation. The data suggest that there is little difference
among the groups of microbes used in this study in terms of sulfide oxidation and improved gold recovery, with some apparent
slight advantage in using the hyper-thermophilic microbes.
D 2003 Elsevier B.V. All rights reserved.
Keywords: Biooxidation; Microbial system; Thermal stress
1. Introduction
Newmont Mining operates a commercial-scale
biooxidation heap for pretreatment of refractory
sulfidic gold ore at its Carlin, Nevada operation.
0304-386X/$ - see front matter D 2003 Elsevier B.V. All rights reserved.
doi:10.1016/S0304-386X(03)00143-9
* Fax: +1-303-683-1944.
E-mail address: j.brierley@worldnet.att.net (J.A. Brierley).
www.elsevier.com/locate/hydromet
Hydrometallurgy 71 (2003) 1319
The process entails use of three aerated heaps, each
containing about 787,400 tonnes of ore for bioox-
idation over a period of 150 days. Following bio-
oxidation, the ore is removed from the heaps and
processed through a conventional mill at a rate of
about 10,200 tonnes/day for gold recovery. The plant
is designated as a Bio-mill process as it is a
combination of a biooxidation-heap pretreatment
and a conventional metallurgical milling (cyanide
leach and carbon recovery of gold) technologies.
The commercial process was developed over 10
years of research and developmental testing of pi-
lot-scale (Brierley and Luinstra, 1993; Brierley et al.,
1995) and large-scale demonstration heaps (Shutey-
McCann et al., 1997; Brierley, 1997). During 2000,
the bio-milling project went into production. The
plant produced 2 tonnes (66,000 oz) of gold in 2000
(Kral, 2001).
The Newmont Mining commercial biooxidation
pretreatment heap is the first of its kind. As such,
specific information related to heap biooxidation of
refractory sulfidic gold ores is not available. Expe-
rience now gained with biooxidation heap pretreat-
ment has demonstrated generation of internal high
temperatures of the heaps at relatively low sulfide-
sulfur concentrations. Beck (1967) reported temper-
atures of 6080 jC in copper leach dumps, and
suggested that one may have to reconsider the role
of microbial activity in leaching operations where
these temperature occur. Conventional mesophilic
biooxidation bacteria die off at the high temperatures
(>45 jC). Thermophilic bacteria, which can exist at
temperatures H45 jC, can oxidize sulfide minerals
in conditions of die off for the mesophilic bacteria.
Potential for use of the thermophilic bacteria, their
advantages, shortcomings and technology for on-site
culturing of the inoculum suspension should be
determined to better define and improve the bioox-
idation-heap technology. Conventional mesophilic,
moderate-thermophilic mineral-oxidizing bacteria,
and hyper-thermophilic archaea were compared for
effect on sulfide oxidation and associated gold
recovery. Effect of temperature on the mineralogical
characteristics was also evaluated.
This report summarizes results of the comparative
study. The results have potential for indicating how
heaps could be better designed, controlled and inoc-
ulated and extend the knowledge base for use of the
bacterial catalysts in the biooxidation-heap pretreat-
ment process.
2. Materials and methods
2.1. Biooxidation pretreatment
Column testing was performed to compare bioox-
idation at different temperatures and effect on subse-
quent gold leaching. Water-jacketed columns and
circulating water baths were set up for temperature
control. Five columns were operated at a different
temperature regime: 2023 jC; 35 jC; 50 jC; 60 jC;
and one column varied from 2023 to 60 jC. For the
variable temperature column, the temperature was
increased stepwise from 2023 jC, to 35 jC, to 50
jC, to 60 jC in 2-week increments; after 2 weeks at
60 jC, the temperature was decreased every 2 weeks
in the same stepwise manner. A thermistor was buried
in the ore of each column for monitoring internal
temperature.
Each column contained 10 kg of sulfidic ore,
crushed to 100% passing 12.7 mm, from Newmonts
Carlin, Nevada Gold Quarry Mine operation. The
Newmont patented pre-inoculation process was per-
formed for each column (Brierley and Hill, 1994). Ore
for each column was mixed with 300 mL of inoculum.
The following inocula were used: (1) columns at 20
23 and 35 jC, conventional laboratory culture of
Acidithiobacillus ferrooxidans and Leptospirillum fer-
rooxidans; (2) column at 50 jC, moderate-thermo-
philic iron-oxidizing Sulfobacillus-type bacteria; (3)
column at 60 jC, hyper-thermophilic archaea, Acid-
ianus and Metallospheara species; and (4) an equal
mix of all of the above for the variable temperature
column.
2.2. Biooxidation monitoring
The circulating biooxidation liquor for each col-
umn was monitored on a weekly basis for pH,
Eh(SCE) and soluble iron concentrations. Column
temperatures were recorded daily, five times per week.
Every 2 weeks, the biooxidation solution was
aseptically sampled at each columns discharge point
to estimate the density of microbial populations. Serial
dilutions were prepared from each sample for incuba-
J.A. Brierley / Hydrometallurgy 71 (2003) 1319 14
tion at 30, 50 and 60 jC. A basal salts solution
containing per liter, 0.4 g (NH
4
)
2
SO
4
, 0.4 g MgSO
4

7H
2
O and 0.04 g KH
2
PO
4
, was used with ferrous iron
energy source (33.3 g FeSO
4
7H
2
O). Dilution cultures
at 50 and 65 jC were supplemented with 0.02% (w/v)
yeast extract to facilitate growth of any thermophiles
present. In addition, the 60 jC dilution cultures were
supplemented with about 0.1 g FeS
2
to enhance
growth of hyper-thermophiles.
2.3. Ore sample characteristics
Table 1 provides the head assay for the Gold Quarry
Mine ore sample used in this study. The ore, containing
3.1 ppm Au/tonne, is sulfidic-refractory. Sulfide con-
tent is 1.8% as pyrite. The size fraction assay for the
sample indicates enrichment of gold in the fine fraction
( 0.295 mm). This gold is unlocked free gold (non-
refractory) as indicated by the 76.6% ratio of cyanide-
soluble gold to total gold determined by fire assay.
Overall, the free gold content is between 42%and 46%,
determined either by assay of a composite sample or
calculated from assay of the size fractions, indicating
the refractory nature of the ore sample.
2.4. Leach procedure
Bottle roll leach procedures were used to determine
the potential gold extraction following biooxidation
pretreatment at each temperature condition. Heap
leaching was simulated using a 96-h bottle roll leach
with the whole ore at minus 12.7 mm. The initial
cyanide concentration was 0.33 g NaCN/L without
carbon. The Bio-mill process was simulated with
biooxidized ore crushed to about P
80
of 0.074 mm and
leached for 24 h with 0.33 g NaCN/L and 15 g
carbon/L.
3. Results and discussion
3.1. Column operation parameters
Weekly column monitoring data are presented in
Figs. 13. Fig. 1 shows the increase in total soluble
iron from biooxidation of the pyrite. The data suggest
biooxidation pretreatment achieved an effective oxi-
dation of the exposed pyrite by about day 60, when
soluble iron levels tended to plateau. This phenom-
enon appeared to be independent of temperature. The
final total soluble iron concentrations may have been
influenced by temperature as the concentrations
ranged from about 21 to 31 g Fe/L. The lowest
soluble iron concentration occurred with the column
run at ambient temperature, even though iron solubi-
lization appeared to have terminated at about day 60.
The data suggest greater iron precipitation at the
higher temperatures.
Fig. 2 compares the solution Eh values with
temperature of incubation. Temperature appears to
influence the redox potential either through physico-
chemical or microbiological causes. The higher tem-
perature columns operated at lower redox values.
Increasing the temperature of the variable temperature
column resulted in lowering of the redox potential
during the 50 and 60 jC periods. Returning the
variable temperature column to 35 jC and ambient
temperature resulted in an increase in redox potential.
Literature reports state the thermophilic microorgan-
isms operate at lower redox, suggesting their iron
oxidation systems require greater concentrations of
ferrous iron than the mesophilic iron-oxidizing bacte-
ria. This would result in a higher ferrous to ferric iron
ratio with concomitant lower Eh.
Fig. 3 presents the comparative solution pH values
for the five columns. The pH of the higher tempera-
Table 1
Gold Quarry Mine ore sample assay
Sample
particle size
Total
Au, ppm
Cyanide-soluble
Au, ppm
Ratio, CN-soluble
Au/total Au 100
Sulfide-S,
%
Fe, %
12.7 + 6.35 mm 2.09 0.51 24.4 1.94 1.69
6.35 + 1.65 mm 2.74 0.99 36.1 1.78 1.67
1.65 + 0.295 mm 4.63 2.98 64.4 1.74 1.69
0.295 mm 7.30 5.59 76.6 2.06 1.85
Calculated 3.09 1.41 45.6 1.83 1.69
Composite 3.12 1.30 41.7 1.82 1.75
J.A. Brierley / Hydrometallurgy 71 (2003) 1319 15
ture columns and the variable temperature column at
higher temperatures tended to be lower. This may
reflect hydrolysis of ferric iron, resulting in acid
production at the higher temperatures. The solution
pH remained in a range favorable for growth of the
mineral-oxidizing bacteria throughout the course of
the experimentation.
3.2. Microbial response to temperature
An aspect of the comparative temperature study
was to monitor the response of microbial populations
to temperature. The viable counts were estimates of
population density of the microbes capable of growth
at either 30, 50 or 60 jC in the circulating biooxida-
tion liquor. Mesophilic bacteria predominated in the
column incubated at ambient room temperature. A
substantial mesophilic population between 10
7
and
10
9
/mL, developed and maintained throughout the
biooxidation period. The ore was not inoculated with
either moderate- or extreme-thermophilic microbes,
but a low level (10/mL) of the moderately thermo-
philic bacteria was detected. The moderately thermo-
philic bacteria are capable of survival and slow
Fig. 2. Solution oxidationreduction potential (SCE) for the comparative biooxidation columns operated at different temperatures.
Fig. 1. Total soluble iron for the comparative biooxidation columns operated at different temperatures.
J.A. Brierley / Hydrometallurgy 71 (2003) 1319 16
growth under mesophilic conditions. No hyper-ther-
mophilic bacteria were detected. Prevailing low tem-
perature conditions for this column test preclude
development of the thermophilic microbes.
Similar results occurred for the microbial popula-
tion response in the column maintained at 35 jC. The
mesophilic bacterial population density was lower in
this column compared with the ambient temperature
column. The mesophiles varied between 10
4
and 10
5
/
mL increasing to 10
9
/mL at the end of biooxidation.
The moderately thermophilic bacteria were not delib-
erately inoculated onto the ore, and they remained at
low numbers (1010
2
/mL). No hyper-thermophilic
microbes were detected.
The moderately thermophilic bacteria were
detected at population densities of up to 10
5
10
6
/
mL for the column operated at 50 jC. Surprisingly,
mesophilic iron-oxidizing bacteria were also present
at high numbers (10
3
10
6
/mL). This suggests either
survival of the mesophiles as they pass through the
column, the reservoir was at ambient temperature, or
growth occurred at locations of the column not at 50
jC. Most likely, the mesophilic bacteria were present
in the column reservoir and not in the column where
the internal thermocouple indicated a consistent 50 jC
temperature. No hyper-thermophilic microbes were
detected in this column.
The hyper-thermophiles were detected in the col-
umn operated at 60 jC. A population density up to
10
6
/mL was present. The other populations of
microbes remained at lower densities (10
2
10
3
/mL
for mesophilic and moderately thermophilic bacteria),
indicating temperature stress on the microbes.
Fig. 4 provides population density data from the
column with variable temperature. The response of the
three groups of microorganisms to variation of tem-
Fig. 4. Comparison of microbial populations capable of growth at respective temperatures in the variable temperature column.
Fig. 3. Solution pH values for the comparative biooxidation columns operated at different temperatures.
J.A. Brierley / Hydrometallurgy 71 (2003) 1319 17
perature is evident. The hyper-thermophiles did not
increase until the temperature was increased to 50 jC
and above. Lowering the temperature decreased the
population density of this group of microbes. The
moderate-thermophilic bacteria increased in number
until the temperature was raised to 60 jC; at that
point, decreasing and remaining stable in number as
further decrease in temperature occurred. Decimation
of the mesophilic iron-oxidizing bacteria occurred
when the temperature was increased to 50 and 60
jC. However, this population increased as the tem-
perature was lowered. The mesophilic bacteria appear
to survive in the ambient temperature column solution
reservoir and subsequently recolonize the ore when
the temperature decreases to a favorable range.
Hyper-thermophiles were only detected in the
columns (60 jC and variable temperature) inoculated
with this group. In contrast, the mesophilic and
moderately thermophilic iron-oxidizing bacteria coex-
ist. A large biooxidation pretreatment facility will
likely require a unit to produce the hyper-thermophilic
archaea for inoculation of the ore to facilitate bioox-
idation under extremely thermophilic conditions.
3.3. Biooxidation of sulfide
Sulfide oxidation data for the respective columns
are presented in Table 2. These data indicate increas-
ing oxidation at elevated column temperatures. There
was little difference in amount of sulfide oxidation at
either ambient room temperature or 35 jC. Increasing
the biooxidation temperature from 35 to 50 jC in-
creased apparent sulfide oxidation from about 35 to
47%. At 60 jC, sulfide oxidation was highest at about
51%. The variable temperature column also had
higher sulfide oxidation, 41%, attributable to periods
of high temperature biooxidation.
3.4. Gold recovery following biooxidation
pretreatment
The resultant gold extractions following biooxida-
tion pretreatment are presented in Table 3. Higher
gold recovery occurred using the Bio-mill CIL
bottle roll leach procedure for pulverized biooxidation
ore compared to the coarse ore bottle roll using
cyanide without carbon. The coarse bottle roll extrac-
tion simulating a heap cyanidation process improved
from the baseline control of 45% to 6872%, the
highest occurring for the 60 jC biooxidized ore.
Pulverizing the biooxidized ore, which simulates the
Bio-mill process, increased gold recovery from a
baseline of 44% to 8595%, the highest recovery for
the 60 jC biooxidized ore.
The Bio-mill process results in higher recovery
of gold following biooxidation pretreatment compared
to gold recovery by a leach process. Another advan-
tage of Bio-milling is more rapid recovery of gold
and capital return. However, a simple heap leach
following biooxidation pretreatment can be used for
lower grade ores.
3.5. Mineralogy of biooxidized residues
A study of possible impact of temperature on
formation of residual iron compounds resulting from
biooxidation was conducted (Chen and Dutrizac,
2000). The hydrated iron sulfate mineral ferricopiapite
[Fe
4.67
3 +
(SO
4
)
6
(OH)
2
20H
2
O] was identified as the pri-
mary iron precipitate in samples from all columns.
Higher temperature favored crystallinity of this min-
eral. An unexpected benefit of biooxidation at temper-
Table 2
Sulfide-S oxidation at respective incubation temperatures
Biooxidation
temperature, jC
Sulfide-S
content, %
Sulfide-S
oxidation, %
2324 1.17 36.1
35 1.13 38.3
50 0.96 47.5
60 0.90 50.8
Variable, 23 !60 !23 1.08 41.0
Table 3
Gold extraction following biooxidation pretreatment
Conditions Biooxidation Gold extraction, %
temperature, jC
CN leach
minus
12.7 mm
CN-CIL
leach minus
0.074 mm
Control not applicable 45.4 43.6
Biooxidized 2324 67.8 88.5
Biooxidized 35 72.8 91.5
Biooxidized 50 68.7 85.0
Biooxidized 60 72.4 95.2
Biooxidized variable,
23 !60 !23
69.2 85.3
J.A. Brierley / Hydrometallurgy 71 (2003) 1319 18
atures z50 jC was an apparent precipitation of
arsenic, probably as scorodite [FeAsO
4
2H
2
O]. Oth-
erwise, there was little difference in terms of conduct-
ing biooxidation at low or high temperatures on
formation of residual iron compounds.
Acknowledgements
The author thanks Newmont Mining, Malozemoff
Technical Facility, 10101 East Dry Creek Road,
Englewood, CO 80112, USA, for permission to
publish this manuscript based on research conducted
while the author was Chief Research Scientist,
Biohydrometallurgy for Newmont Mining.
References
Beck, J., 1967. The role of bacteria in copper mining operations.
Biotechnology and Bioengineering 9, 487497.
Brierley, J., 1997. Heap leaching of gold bearing deposits, theory
and operational description. In: Rawlings, D.E. (Ed.), Biomin-
ing: Theory, Microbes and Industrial Processes, Springer-Verlag
and Landes Bioscience, Georgetown, TX, pp. 103115.
Brierley, J.A., Hill, D.L., 1994. Biooxidation process for recovery
of metal values from sulphur-containing ore materials. US Pat-
ent No. 5,332,559.
Brierley, J.A., Luinstra, L., 1993. Biooxidation-heap concept for
pretreatment of refractory gold ore. In: Torma, A.E., Wey, J.E.,
Lakshmanan, V.I. (Eds.), Biohydrometallurgical Technologies.
Bioleaching Processes, vol. I. The Minerals, Metals and Materi-
als Society (TMS), Warrendale, PA, pp. 437448.
Brierley, J.A., Wan, R.Y., Hill, D.L., Logan, T.C., 1995. Biooxida-
tion-heap pretreatment technology for processing lower grade
refractory gold ores. In: Vargas, T., Jerez, C.A., Wiertz, J.V.,
Toledo, H. (Eds.), Biohydrometallurgical Process, vol. I. Uni-
versity of Chile, Santiago, pp. 253262.
Chen, T.T., Dutrizac, J.E., 2000. Identification of iron precipitates in
biooxidized gold ore. CANMET Report MMSL 2000-048, Ot-
tawa, 2000.
Kral, S., 2001. Mining at Deep PostNewmonts newest under-
ground mine. Mining Engineering 53 (12), 2529.
Shutey-McCann, M.L., Sawyer, F.-P., Logan, T., Schindler, A.J.,
Perry, R.M., 1997. Operation of Newmonts biooxidation dem-
onstration facility. In: Hausen, D.M. (Ed.), Global Exploitation
of Heap Leachable Gold Deposits. The Minerals, Metals and
Materials Society (TMS), Warrendale, PA, pp. 7582.
J.A. Brierley / Hydrometallurgy 71 (2003) 1319 19