J4J

The Luropean |ournal of Nineral Processing and Lnvironmental Protection

Vol.2, Mo.3, J3D3-DB6B, 2DD2, pp. J4J-J5D
Lffect of Salinity on the Cxidative Pctivity of Pcidophilic acteria
during ioleaching of a Complex Zn]Pb Sulphide Cre

H. Deveci*

KaraUcn!z Tccnn!ca1 Un!+crs!1], A!n!ng ng!nccr!ng Ocpar1ncn1, UOBO TraDzcn, TDrRc]

Received 3 Ppril 2DD2 accepted JD |une 2DD2

PSTRPCT

The presence of some anions and cations at certain levels in the bioleaching environment may exert an
inhibitory effect on the grovth and hence leaching activity of a bacterial culture. In this respect, the quality
of process vater available vith particular reference to salinity can be of prime importance for the
application or development of a bioleaching process for a particular feed at an operation site.
The current study investigates the extent to vhich salinity up to B% Cl
-
(BDg]l) affects the bioleaching
activity of mesophilic, moderately and extremely thermophilic strains of bacteria during the bioleaching of a
complex Zn]Pb sulphide ore. The results indicated that salinity can adversely influence the "optimum"
bioleaching activity of mesophiles and moderate thermophiles; the extent being dependent upon the strain
(and type) of bacteria and the concentration of chloride. The mesophilic W|N strain vas found to oxidise the
complex ore at concentrations of up to D.B% Cl
-
(Bg]l) vithout any significant effect on the extraction of
zinc vhile the limited extraction of zinc by DSN 5B3 strain occurred at D.2% Cl
-
. It vas noted that
mesophiles can be adapted to tolerate D.B-J% Cl
-
(B-JDg]l) in solution. The bioleaching ability of the
strains of moderate themophiles vas adversely influenced even at D.2% Cl
-
(2g]l). Cn the other hand, the
extreme thermophiles vere shovn to perform vell under saline conditions up to 5% Cl
-
(5Dg]l). This
probably indicates the halophilic peculiarity of the extreme thermophiles compared vith the mesophiles
and the moderate thermophiles. 2DD2 SDU. Pll rights reserved.

Keyvords. Pcidophilic bacteria; ioleaching; Chloride

J. IMTRCDUCTICM

iooxidation of gold bearing arsenopyrite]pyrite concentrates using mesophilic and
moderately thermophilic bacteria as a pre-treatment step prior to cyanidation has already been
proved as an economically viable and competitive alternative to conventional roasting and
pressure oxidation (Dev c1 a1., J997; Niller, J997). This has stimulated the extension of the
technology for the extraction of other metals such as Cu, Zn, Co and Mi (rierley and rierley,
J999; Niller c1 a1., J999; Gilbertson, 2DDD).
Iron and]or sulphur oxidising strains of mesophilic bacteria such as Tn!cDac!11Ds /crrcc!Uans,
Lcp1cspr!11Dn /crrcc!Uans and Tn!cDac!11Ds 1n!cc!Uans operating at 4D
o
C are the most
extensively studied microorganisms for the oxidation sulphide minerals vith commercial
interest (Dev c1 a1., J997; |ordan c1 a1., J996). Since the kinetics of metal dissolution tend to
increase vith temperature, the commercial application of thermophilic bacteria vhich can
operate at temperatures exceeding 45
o
C has great potential for improving the rates of metal
extraction from sulphide minerals. Pccordingly, in recent years several commercial bioleaching
processes using thermophilic bacteria have been developed for the extraction of base metals
(Cu, Mi and Zn) and successfully operated at pilot scale vith the reports of imminent
commercial realisation (Niller c1 a1., J999; Staden, J99B; Gilbertson, 2DDD).

* Corresponding author. L-mail. hdeveci@ktu.edu.tr
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H. Deveci ] The Luropean |ournal of Nineral Processing and Lnvironmental Protection
Vol.2, Mo.3, J3D3-DB6B, 2DD2, pp. J4J-J5D

ioleaching of the sulphide minerals is based on the exploitation of the ability of the
acidophilic bacteria to oxidise ferrous iron (Lq. J) and]or reduced sulphur compounds (Lq. 2 for
elemental sulphur) as substrates vhereby the bacteria derive the energy required for the grovth
and other metabolic functions (Ingledev, J9B2). The oxidation of ferrous iron and sulphur
compounds by bacteria results in the production of ferric iron and]or acid vhich attack sulphide
minerals (Lqs 3-4 for ZnS) (Sand c1 a1., 2DDJ). This leads to the enhanced dissolution of metals
from sulphides in the presence of bacteria.

2FeSC
4
+ J]2C
2
+ H
2
SC
4
bacteria
 o  Fe
2
(SC
4
)
3
+ H
2
C (J)
S
o
+ 3]2C
2
+ H
2
C
bacteria
 o  H
2
SC
4
(2)
ZnS + J]2C
2
+ H
2
SC
4
o ZnSC
4
+ H
2
C + S
o
(3)
ZnS + Fe
2
(SC
4
)
3
o ZnSC
4
+ 2FeSC
4
+ S
o
(4)

The degree of enhancement in the metal dissolution is intimately controlled by the oxidising
activity (grovth) of bacteria. These microorganisms in general obtain their cellular material from
inorganic sources such as CC
2
, MH
4
+
, K
+
, Ng
2+
, Ca
2+
, SC
4
2-
and PC
4
3-
present in the grovth
media (Tuovinen and hatti, J999). Hovever, the presence of some anions such as Cl
-
and
cations at certain levels in the bioleaching environment may exert an inhibitory effect on the
grovth and hence the leaching performance of a bacterial culture. In this regard, the quality of
process vater available vith particular reference to salinity can be of prime importance for the
application of the bioleaching processes since chloride-free process vater may not be available
at an operation site. udden and Spencer (J99J) and Weston c1 a1. (J994) have reported the
analyses of some borehole vaters vith a Cl
-
content of J2.7-25.7g]l and JJDg]l respectively.
Depending on the concentration of chloride that a bacterial culture can tolerate the treatment of
poor quality vater may be required, vhich vould in turn lead to the increase in both operating
and capital costs (udden and Spencer, J99J).
Leong c1 a1. (J993) reported that the dissolution of copper from a high grade ore by a
mesophilic mixed culture of Tn!cDac!11! vas adversely affected in the presence of up to Bg]l Cl
-
.
Weston c1 a1. (J994) observed a reduction in the oxidation rate of pyrite vith an increase in the
concentration of Cl
-
above 2g]l during the pilot scale test vork on a pyritic gold concentrate
using the ICX

culture. udden and Spencer (J99J) claimed that moderately thermophilic

bacteria (acTech

`s culture) can operate vell under saline conditions (D-5% MaCl).

The current study reports the results of an investigation undertaken to evaluate the Cl
-
tolerance limits of all three groups of bacteria; mesophiles, moderate and extreme thermophiles
during the bioleaching of a complex ore.

2. LXPLRINLMTPL

2.J. Nineralogy and Chemical Composition of the Cre

Samples of the NcPrthur River complex ore vere obtained from the mine site in the
Morthern Territory of Pustralia (kindly via NIN Holdings, ovners of the deposit). The chemical
and mineralogical analyses of the ore are shovn in Table J. Nineralogical analysis of the ore
using the XRD and SLN revealed that the ore consisted predominantly of sphalerite (ZnS),
galena (PbS) and pyrite (FeS
2
) as the important sulphide phases. The sphalerite, galena and
pyrite vere found to be very fine grained and intimately associated vith each other to such a
degree that even fine grinding (<JDm) vould not have produced complete liberation. P
particle size of <3DDm (d
BD
= -25Dm) vere used in the experiments.

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H. Deveci ] The Luropean |ournal of Nineral Processing and Lnvironmental Protection
Vol.2, Mo.3, J3D3-DB6B, 2DD2, pp. J4J-J5D

Table J
Chemical and mineralogical composition of the NcPrthur River complex ore used in this study
Netal content Nineralogical content
Zn
(%)
Fe
(%)
Pb
(%)
Cu
(%)
S
(%)
Pg
(g]t)
J6.2 7.95 5.6D D.27 J5.2 59
Sphalerite, galena, pyrite,
chalcopyrite, quartz, dolomite and
various silicate phases such as
ortochlase
2.2. acterial Cultures

Tvo strains of mesophiles, moderate and extreme thermophiles vere used in this study. The
mesophiles vere Tn!cDac!11Ds /crrcc!Uans DSN 5B3 (from Deutsche Sammlung von
Nikroorganismen (DSN) culture collection) and the mixed enrichment culture designated W|N
(an isolate from the acid mine drainage vaters of Wheal |ane mine, Cornvall, UK) composed of
iron and sulphur oxidisers such as Tn!cDac!11Ds /crrcc!Uans, and heterotrophic bacteria. The
strains of moderate thermophiles vere YTFJ and THWX (5D1/cDac!11Ds ac!Ucpn!1Ds) vhich vere
kindly provided by Dr D.. |ohnson (University of Wales, angor, UK). The extreme thermophiles,
DSN J65J (c!U!anDs Br!cr1c]!) and 5D1/c1cDDs vere available in the culture collection of
Camborne School of Nines and had previously been obtained from the DSN culture collection
and kindly from Dr. P.R. Morris (Department of iological Sciences, Warvick University, UK)
respectively. Pll cultures vere grovn (and maintained) on the ore sample (J% v]v) prior to the
experimental vork. The grovth of the cultures vas carried out in an enriched salt solution
containing NgSC
4
.7H
2
C (D.4g]l), (MH
4
)
2
SC
4
(D.2g]l), K
2
HPC
4
.3H
2
C (D.Jg]l) and KCl (D.Jg]l).
Yeast extract (D.D2% v]v) vas also added to support the grovth of moderate thermophiles.

2.3. ioleaching experiments

ioleaching experiments (in duplicate) vere carried out in 25Dml Lrlenmeyer flasks. Lnriched
salt solution (9Dml) vas transferred into each flask to vhich Jg of the ore sample (J% v]v pulp
density) vas added. The calculated (stoichiometric) amount of chloride (as MaCl) vas added to
each flask to produce the required chloride concentration in the final volume of solution. The
flasks vere then autoclaved under the pressure of Jatm at J2J
o
C for 2D minutes. Folloving
autoclaving each flask vas inoculated under aseptic conditions vith a JDml aliquot of the
selected culture producing a final volume of slurry of approximately JDDml. The control flasks
vere prepared using the same procedure vith the exception that, instead of being inoculated
vith a bacterial culture, JDml of 5% thymol in methanol vas transferred to prevent any
bacterial activity for the duration of experiments. For the experiments using the mesophiles an
orbital shaker operating at JDDrpm vithin a varm room controlled to a temperature of 3DqC
vas used to shake the flasks. The moderate and extreme thermophile experiments vere
performed using a separate incubator rotating at J6Drpm, thermostated at the appropriate
temperature such as 5DqC for moderate thermophiles and 7D
o
C for extreme thermophiles.
Sampling of each flask vas carried out daily by removing a Jml aliquot of the leach solution
vhich vas then used for analysis of metals (Zn, Fe and Pb) by Ptomic Pbsorption Spectrometry
(PPS) and for monitoring of pH. Pd[ustment of the pH vas undertaken only if the pH value
exceeded the initial pre-set values of pH J.6 for mesophiles and moderate thermophiles, and
pH J.4 for extreme thermophiles.

3. RLSULTS PMD DISCUSSICM

3.J. Lffect of Chloride on Nesophiles

Figure J-4 illustrate the dissolution of zinc and iron from the complex ore using mesophilic
strains W|N and DSN 5B3 in a range of D-5% Cl
-
added. In the presence of strains (D% Cl
-
) there
vas a significant enhancement in the extraction of zinc and iron in comparison vith the chemical
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H. Deveci ] The Luropean |ournal of Nineral Processing and Lnvironmental Protection
Vol.2, Mo.3, J3D3-DB6B, 2DD2, pp. J4J-J5D

acid leaching in the absence of strains as indicated by the control. The results also indicated that
the addition of chloride exerted an adverse effect on the oxidative activity of the bacterial cultures
vith the extent being dependent upon the strain and concentration of chloride added.
Ps shovn in Figure J-2 for W|N strain, the addition of chloride led to a lag period in the
dissolution of zinc and iron vith the extent of the lag period having increased vith increasing the
concentration of chloride. This increase in the lag period vas possibly the indication of the adverse
effect of chloride on the bioleaching activity of the strain. During these initial periods, the
adaptation of the strain or selection of the more resilient cells vithin the bacterial population may
be presumed to have occurred since a substantial increase in the dissolution of zinc and iron
occurred particularly at Cl
-
concentrations of dD.B% folloving these initial periods.

Figure J. Lffect of chloride on the extraction of zinc from the ore (J% v]v) by W|N strain at pH J.6 and
3D
o
C
Figure 2. Lffect of chloride on the extraction of iron from the ore (J% v]v) by W|N strain at pH J.6 and 3D
o
C
The extraction of zinc vas comparable at dD.B% Cl
-
vith a slight decrease at D.5-D.B% Cl
-
over
the bioleaching period (Figure J). Further increase in the Cl
-
level resulted in significant reduction in
the rate and extent of metal dissolution. The extraction of zinc at 3-5% Cl
-
appeared to be
prematurely levelled off after JB9h. This could be attributed to the formation]encrustation of a
passivating layer such as elemental sulphur and]or ferric iron precipitates on the surfaces of
sphalerite particles. In this respect, the presence of relatively high concentrations of Ma
+
such as
2D g]l at 3% Cl
-
(introduced in the form of MaCl) could have promoted the precipitation of sodium
[arosite (Lq. 5) providing that the bacteria vere able to generate ferric iron under these conditions.

Ma
+
+ 3Fe
3+
+ 2SC
4
2-
+ 6H
2
C o MaFe
3
(SC
4
)
2
(CH)
6
+ 6H
+
(5)
0
20
40
60
80
100
0 100 200 300
Time in hours
ControI
(0)
ControI
(5)
0 CI
0.2
0.5
0.8
1
3
0
20
40
60
80
100
0 100 200 300
Time in hours
0 CI
0.2
0.5
0.8
1
3
5
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H. Deveci ] The Luropean |ournal of Nineral Processing and Lnvironmental Protection
Vol.2, Mo.3, J3D3-DB6B, 2DD2, pp. J4J-J5D

The onset of a substantial dissolution of iron vas coincident vith the occurrence of a significant
extraction of zinc, 45% Zn at D% Cl
-
(Figure J-2). This vas probably a reflection of the
electrochemical aspects of the dissolution process peculiar to the complex sulphide systems vith
"preferential or selective" oxidation of sphalerite over pyrite (Matara[an, J99D). The continual
oxidation of ferrous iron by bacteria vas presumed to occur vithin the bioleaching environment.
This vould probably lead to the gradual build-up of ferric iron as a consequence of depletion of
more active sulphides such as sphalerite and, more importantly, proliferation of the bacterial
population vith increasing oxidation capacity. Had the redox potential of the system
approached]surmounted the rest potential of the pyrite, the dissolution of iron vould have been
promoted. This vas consistent vith the observed pattern for the dissolution of iron (Figure 2).
Figure 3-4 illustrate the effect of added Cl
-
on the bioleaching performance of DSN 5B3 strain.
Compared vith W|N strain, DSN 5B3 strain appeared to be more sensitive to Cl
-
ions in that
severe inhibition of the oxidising activity of the strain vas discernible even at D.2% Cl
-
vith a J7%
decrease in the final extraction of zinc. This highlights the different sensitivity of both strains to
tolerate Cl
-
ions. Pn apparent decrease in the concentration of iron in solution at D.2% Cl
-
after
26Dh vas observed for DSN 5B3 strain. This could be attributed to the formation of presumably
[arosite type precipitates at the operating pH of J.6. In this regard, the limited zinc extraction at
D.2% Cl
-
may have been due to the encrustation of precipitates on the surfaces of sphalerite
particles possibly leading to the impediment of the further zinc release.
The dissolution of lead vithin the experiments vhere no chloride vas added vas minimal; over
9B% of lead reported to the residues most likely in the form of either "insoluble" lead sulphate or
undissolved galena irrespective of the strain (and type) of bacteria used (the solubility of PbSC
4
is
D.D45g]l at 25
o
C (Forvard and Peters, J9B5)). Hovever, the solubilisation of lead vas promoted
vith increasing the concentration of Cl
-
added such as 5% Pb at J% Cl
-
and JDD% Pb (D.44g]l) at
5% Cl
-
. It should be noted that lead chloride is reasonably soluble (the solubility of PbCl
2
is JD.7g]l
at 25
o
C (Forvard and Peters, J9B5)) and the solubility of lead increases vith increasing Cl
-
concentration and temperature probably due to the formation of soluble lead-chloro complexes,
PbCl
i
2-i
vhere i=J-4 (Holdich and Lavson, J9B7).
Folloving the termination of the experiments, the cultures from the flasks at each Cl
-
level vere
then subcultured on ferrous iron (JDDmN, pH J.7) and the oxidation of ferrous iron vas observed
to occur in the flasks vhich had received inoculum from the flasks containing up to D.5% Cl
-
for
DSN 5B3 strain and up to J% Cl
-
for W|N strain. It vas noted that a prolonged period for the
oxidation of ferrous iron vas necessary for the strains vhich had been exposed to the increasing
concentrations of Cl
-
. Pdaptation of both strains to Cl
-
vas also carried out on ferrous iron using
serial subculturing procedures. Plthough the oxidation of ferrous iron vas relatively slov, both
strains vere developed to tolerate D.B% and J% Cl
-
(B and JDg]l) respectively beyond vhich
there vas little success.

Figure 3. Lffect of chloride on the extraction of zinc from the ore (J% v]v) by DSN 5B3 strain at pH J.6 and
3D
o
C
0
20
40
60
80
100
0 100 200 300
Time in hours
ControI (0)
0 CI
0.2
0.5
0.8
1
3
5
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H. Deveci ] The Luropean |ournal of Nineral Processing and Lnvironmental Protection
Vol.2, Mo.3, J3D3-DB6B, 2DD2, pp. J4J-J5D

Figure 4. Lffect of chloride on the extraction of iron from the ore (J% v]v) by DSN 5B3 strain at pH J.6 and
3D
o
C
Leong c1 a1. (J993) observed the detrimental effect of chloride (D-Bg]l Cl
-
) on the dissolution
of copper using mixed cultures of Thiobacilli. These investigators also shoved the adaptation of
bacteria to chloride levels of 5g]l ameliorating the deleterious effect of chloride on the
dissolution process. Vorreiter and Nadgvick (J9B2) demonstrated that the average release of
copper decreased (by BD%) in the presence of 2Jg]l Cl
-
(as MaCl) during the bioleaching of a
lov grade copper ore using T. /crrcc!Uans in a percolation column. They also noted a decrease
in the number of viable cells (T. /crrcc!Uans) from JD
3
to <JD cells]ml on average.
Weston c1 a1. (J994) reported the results of pilot plant and bench scale test vork carried out
using the ICX

process for the biooxidation of a refractory gold concentrate vith no apparent

effect of salinity at J-2g]l Cl
-
on the bacterial oxidation rate vhen operating at lover pHs (J.J-
J.3). They argued that the deleterious effect of salt vas not due to the chloride present but
rather due to the coexisting ions such as sodium leading to the formation of [arosite precipitates.
They also reported a significant reduction in the subsequent extraction of gold by cyanidation at
high Cl
-
levels vhich vas attributed to the increasing amount of [arosite precipitates under these
conditions. Dev c1 a1. (J997) suggested that high concentrations of chloride in solution could
damage the membrane of the bacteria. They also reported that the negative effect of chloride
on the oxidation of ferrous iron by the ICX

culture vas negligible in a range of D-5g]l Cl

-
,
thereafter the oxidation rate of ferrous iron significantly decreased by 25% at 7g]l Cl
-
vith
complete inhibition of the activity of bacteria at Cl
-
concentrations of tJ9g]l.
It can be inferred that although the salinity exerts an adverse effect on the oxidising activity
of mesophiles depending on the concentration of Cl
-
and the strain of bacteria, the adaptation of
the mesophilic bacteria up to D.B-J% Cl
-
(B-JDg]l) appears to be possible. Hovever, the
presence of the counter ions such as sodium may present further problems partaking in the
formation of potentially deleterious [arosites.

3.2. Lffect of Chloride on Noderate Thermophiles

The addition of chloride severely impeded the oxidative activity of YTFJ strain even at the
lovest Cl
-
level tested (D.2%) as illustrated in Figure 5. The strain completely solubilised the zinc
available in the ore in the absence of added Cl
-
(D% Cl
-
). Hovever, a severe decrease in the rate
and extent of zinc extraction vas observed in response to the addition of chloride at D.2-B% Cl
-
vhere a limited extraction of zinc occurred over the bioleaching period.
Cn the contrary, previous studies by the author (Deveci, J997) had demonstrated that the
addition of chloride (J-4% Cl
-
) had no apparent effect on the dissolution of zinc by YTFJ strain
from the ore but all be at a slover rate (at D% Cl 9.5mg Zn]l]h compared vith J6.5mg Zn]l]h
in the current study). These findings vere later confirmed using the same strain at D-3% Cl
-
(Deveci, 2DDJ). These earlier tests (Deveci, J997) had been performed in the absence of yeast
0
20
40
60
80
100
0 100 200 300
Time in hours
0 CI
0.2
0.5
0.8
1
3
5
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extract. The limited ability of the strain for the oxidation of ferrous iron had been noted and
attributed to the maintenance of the strain in the absence of yeast extract. It should be noted
that many pure strains of moderate thermophiles including those used in this study require
yeast extract to support the grovth of these microorganisms (Deveci, 2DDJ; Ghauri and
|ohnson, J99J). The results of the current tests (at D% Cl
-
) shoved that the dissolution rate of
zinc vas enhanced J.7-fold in the presence of D.D2% v]v yeast extract compared vith the
earlier studies. The rationale behind this behaviour of YTFJ strain maintained in the presence
and absence of yeast extract under saline conditions vas not understood.

Figure 5. Lffect of chloride on the extraction of zinc from the ore (J% v]v) by YTFJ strain at pH J.6, 5D
o
C
and D.D2% v]v yeast extract

The effect of chloride on the bioleaching activity of THWX strain vas also investigated as
shovn in Figure 6. The dissolution of zinc at D.2% Cl
-
vas significantly faster (3.B times) than
that produced by YTFJ strain (Figure 5) at the same level of chloride. This probably indicates the
varying sensitivity of both strains to chloride ions. The extraction of zinc by THWX strain though
vas severely suppressed at D.5-3% Cl
-
(Figure 6).

Figure 6. Lffect of chloride on the extraction of zinc from the ore (J% v]v) by THWX strain at pH J.6, 5DoC
and D.D2% v]v yeast extract

Plthough there appeared a significant impact on the bioleaching ability of the strains used in
this study even at D.2% Cl
-
, udden and Spencer (J99J) reported that the acTech

`s
moderately thermophilic culture vas able to oxidise a pyrite concentrate at a rate comparable
(25% decrease) to the control (D% Cl
-
) in a range of D.6-3% Cl
-
. They attributed the observed
difference in the oxidation rate to the precipitation of iron rather than the inhibition of the
0
20
40
60
80
100
0 50 100 150 200 250
Time in hours
ControI (0)
ControI (8)
0 CI
0.2
0.5
1
3
5
8
0
20
40
60
80
100
0 50 100 150
Time in hours
ControI (0)
0 CI
0.2
0.5
0.8
1
3
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H. Deveci ] The Luropean |ournal of Nineral Processing and Lnvironmental Protection
Vol.2, Mo.3, J3D3-DB6B, 2DD2, pp. J4J-J5D

bacterial activity since their microscopic examination of the culture revealed no apparent
difference betveen the control and the chloride tests. Mo change in the number of bacteria or
the morphology of the cells or the motility of the bacteria vas noted in the presence of salt
(MaCl). In a separate study, udden and Spencer (J993) also demonstrated that the extraction
rate of copper from a chalcopyrite concentrate decreased by 2D% in a range of D.3-J% Cl
-
(added as MaCl) and by 5D% at J.2% Cl
-
. These investigators noted an initial lag period; the
extent being proportional to the amount of chloride present. Hovever, in the subsequent cycles
of bioleaching process, they no longer observed the lag time that vas attributed to the
adaptation of the culture to the saline conditions.

3.3. Lffect of Chloride on Lxtreme Thermophiles

Complete dissolution of zinc vas achieved by 5D1/c1cDDs at all the chloride concentrations
tested (D-5% Cl
-
) over the bioleaching period (Figure 7). The addition of chloride (D.2-5% Cl
-
)
appeared to have a slightly positive effect on the dissolution of zinc compared vith no addition
of chloride. P similar pattern for metal dissolution under the saline conditions vas also
produced by DSN J65J strain as shovn in Figure B. These findings are consistent vith the
earlier findings using both strains at J-4% Cl
-
(Deveci, J997).
The addition of chloride in the range of D-3% Cl
-
also appeared to improve the dissolution of
zinc by DSN J65J strain particularly after the initial 63h (Figure B). The extraction of zinc at 5%
Cl
-
vas similar to that vith no chloride addition during the initial period of J39h despite the
relatively lov zinc recovery over the bioleaching period. Further increase in the concentration
of chloride (at B% Cl
-
) adversely affected the bioleaching process and the final extraction of zinc
vas limited to only 77%.

Figure 7. Lffect of chloride on the extraction of zinc from the ore (J% v]v) by 5D1/c1cDDs strain at pH J.4 and
7D
o
C
It should be noted that there vas no evidence for the precipitation of iron (presumably due
to the relatively lov operating pH of J.4) in the current experiments using the extreme
thermophiles. Motvithstanding this, the precipitation of ferric iron as [arosites vould probably
be promoted by the increase in temperature but this may be partially alleviated by decreasing
pH (Dutrizac, J9B3). The extreme thermophiles can efficiently operate at high levels of acidity
as lov as pH J (Deveci, 2DDJ).
These findings indicate that the extreme thermophiles are halophilic in character as these
microorganisms are able to operate under extremely saline environments at chloride
concentrations up to 5% Cl
-
(5Dg]l).

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H. Deveci ] The Luropean |ournal of Nineral Processing and Lnvironmental Protection
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Figure B. Lffect of chloride on the extraction of zinc from the ore (J% v]v) by DSN J65J strain at pH J.4
and 7D
o
C
4. CCMCLUSICMS

Investigations into the chloride tolerance of the selected strains of mesophiles, moderate and
extreme thermophiles shoved that the quality of process vater vith respect to salinity is of
prime importance for the bioleaching processes since it affects the activity of the bacteria and
the chemistry of the dissolution process. The folloving conclusions can be dravn from this
study.

i) The salinity exerts an adverse effect on the bioleaching activity of the mesophiles and
moderate thermophiles resulting in a decrease in the rate and extent of zinc extraction from
the ore. The extent of the adverse effect depends on the concentration of chloride and the
strain of bacteria used.
ii) The adaptation of mesophilic bacteria to D.B-J% Cl
-
(B-JDg]l) appears to be possible.
Chloride tolerance of moderate thermophiles appears to be limited to a Cl
-
concentration of
D.2% (2g]l).
iii) The extreme thermophiles are halophilic in character vith their ability to perform under
extremely saline environments at Cl
-
concentrations up to 5% (5Dg]l).
iv) The presence of counter ions such as Ma
+
may present further problems as they could
promote the formation of potentially deleterious ferric precipitates.
v) The solubilisation of lead increases vith increasing the concentration of Cl
-
probably due to
the formation of lead-chloro complexes. In the absence of added Cl
-
>9B% of the lead
reports to the residues.

This vork highlights the vital need to consider the quality of process vater available at the
early stages of the development of bioleaching proposals and the potential benefit of using
extreme thermophiles rather than mesophiles and moderate thermophiles for the bioleaching of
ores]concentrates in saline environments.

PCKMCWLLDGLNLMTS

The author vould like to express his sincere thanks and appreciation to the late Dr. C.V.
Phillips for his support, assistance and guidance, and to the Turkish Ninistry of Lducation for
financial support during his studies at Camborne School of Nines, University of Lxeter, UK.

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RLFLRLMCLS

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