ZA200600484B - Cyclic derivatives as modulators of chemokine receptor activity - Google Patents

Description

TITLE

“Improved Bacterial Oxidation of Sulphide Ores and Concentrates”

FIELD OF THE INVENTION

The present invention relates to the improved bacterial oxidation of sulphide ores and concentrates using a bacterial culture.

The bacterial oxidation process of the present invention has particular application in the bacterial oxidation of ores and concentrates containing chalcopyrite.

BACKGROUND ART

Bacterial oxidation has been used for a number of years in successfully processing arsenopyrite, pyrite, pyrrhotite, covellite and chalcocite ores and concentrates, the one exception to this processing has been the oxidation of chalcopyrite (CuFeS;) ores and concentrates.

Prior art mixes of bacteria used to facilitate oxidation of sulphide ores and concentrates, other than chalcopyrite ores and concentrates, use a variety of suites of bacteria. For example, the mixed bacterial culture employed by Gencor

Limited of South Africa comprise predominantly Thiobacillus ferrooxidans,

Thiobacillus thiooxidans and: Leptospirillum ferrooxidans. The Gencor cultures consist of a mixed population of mesophilic bacteria, which operate in the temperature range of 350 C to 45° C (Dew & Miller, 1997). :

Further, Finnish Patent Application 953488 to Gencor Limited discloses the use of

Thiobacillus ferrooxidans, Thiobacillus thiooxidans and Leptospirillum ferrooxidans to achieve oxidation at a pH of preferably 3 with an ore preferably crushed to below 6 mm.

The bacterial culture utilised by BacTech (Australia) Pty Ltd, see for example US

Patent 5429659, is a moderately thermophilic bacterial culture operating in the semperature range of 46° C to 50° C. The cuiture has been designated “M4” ty

Barrett et al (1988) and has been described by Nobar et ai. (1988) (Brierley and

Brans 1994).

The MINBAC process developed by Mintek — Anglo American Corporation based in Randburg, South Africa utilises a mesophilic mixed bacterial culture comprising

Thiobacillus ferrooxidans/Leptospirillum ferrooxidans (Brierley and Brans 1994).

The bacterial cultures presently used are unable to produce commercially acceptable results for chalcopyrite without either ultra fine milling (Pgo<20 um) of the ore or concentrate to facilitate bacterial oxidation, or the use of very long leach times to achieve oxidation. Times of over 100 days are not uncommon.

Current trends are moving towards the use of higher temperatures to encourage ferric oxidation. However, the high temperatures employed lead to having to cool post-oxidation and to provide reactors formed of specialised materials, for example surgical grade stainless steel. Both circumstances increase the cost of such an operation.

Heap leaching is by far the most commonly utilised bacterial process for recovering copper from the more easily oxidised secondary copper minerals such as covellite and chalcocite. The process involves stacking crushed ore onto a specially prepared impermeable pad. The pad is designed so that the pregnant liquor draining from the heap collects at a point from which it is drained to a collection pond. Metals are recovered from the pregnant liquor solution either via precipitation, solvent extraction and/or electrowinning.

In order for successful heap leaching to take place it is essential to maintain the integrity of the heap. The main factor determining the heaps stability is the crush size of the ore. Crushing of the ore must take place to an extent where the ore is fine enough to allow good lixiviant percolation through the heap without excessive channelling taking place whilst also maintaining void spaces essential for good air dispersion and lixiviant drainage. If the ore is crushed too finely percolation through the heap may be very slow. Insufficient void spaces will be present and

. =o inefficient drainage of the heap will occur resulting in pooling in the heap anc a high phreatic head. If on the cther hand the ore size is too coarse, drainage of the heap will be fast and the level of metals in solution will be low, in addition the structure of the heap may fail as the ore is broken down through biological and chemical processes. in many cases the crushed ore is agglomerated with: binders, sulphuric acid and water prior to stacking, the result being a more uniform particle size and acid distribution throughout the heap. oo

Prior to stacking the heap a drainage layer is usually placed on the pad, this is generally composed of unreactive rock such as quartzite and ensures adequate drainage of the pregnant liquor. Heaps are irrigated with acidified bacterial liquor which acts as the lixiviant for leaching of the copper from the ore. The bacteria employed in heap leaching are generally aerobic and therefore require oxygen.

This may be forced into the heap by means of low pressure blowers or air may be sucked into the heap due to a chimney effect that occurs as bacteria oxidise ore and create heat. . The Geocoat process (see WO 96/38381) is a variation on heap leaching and has been marketed by US company Geobiotics. The process involves producing a concentrate from the sulphidic ore, coating this onto crushed, sized rock and ) producing a heap that can be subjected to bacterial oxidation. Importantly, the invention of this process appears to be an effort to ensure adequate air and fluid flow, and heat dissipation, in the heap.

Dump leaching is very similar to heap leaching and is generally reserved for lower grade ores. Often dump leaching will be considered an as accompanying process to heap leaching rather than a stand alone project in its own right. Essentialiy, where waste or low grade rock is to be mined and stockpiled anyway, with little ground preparation beforehand, some value can be extracted from the material. indigenous bacteria will bé present in the heap and all that is required is to promote their activity. This is done by the addition of acid and nutrients to the irrigation solution, such as with heap leaching. The difference is in the cost.

Little or no crushing wiil be performed prior to stacking. Only the bare minimum cf pad preparation is performed and there is no forced aeration.

Vat leaching can be considered to be intermediate between heap leaching and tank leaching in terms of cost, sophistication and efficiency. It is a process in which the material to be treated is fully immersed in the leach solution but is not agitated, at least not to any significant extent, though some agitation due to the air : - and/or solution flow may take place. The process has the advantage over heap or dump leaching in that complete wetting of the mineral surfaces is achieved and channelling is avoided. Finer crush sizes can also be handled better in a vat, though there is still a limit to the fineness imposed by the need for permeability by both the air and solution. Beyond this limit, it becomes necessary to suspend the material in the solution. If the vats are to be single use only, they can be constructed as lined dams, sloped to one corner to allow circulation and recovery of the leach liquor. Multiple use vats would need to be of a more robust construction such as concrete or brick. Aeration would be by submerged pipe or could otherwise by accomplished by intermittently draining the vat and allowing air to be drawn into the ore by the retreating liquor. ~ Tank leaching, as the name suggests, entails the bacterial leaching of aerated mineral slurries in agitated tanks. It is envisaged that the technology would be very similar for base metal bio-leaching, but to date a system for copper has not been commercially developed. :

Available results indicate that the costs associated with ultra-fine milling of the concentrate (Pg <30 um) can be expected to make the capital and operating costs too high.

The process of the present has as one object thereof to overcome the abovementioned problems associated with the prior art, or to at least provide a useful alternative thereto.

The preceding discussion of the background art is intended to facilitate an understanding of the present invention only. It should be appreciated that the discussion is not an acknowledgement or admission that any of the material referred to was part of the common general knowledge as at the priority cate of the application.

Throughout this specification, unless the context requires otherwise, the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

Throughout the specification, reference to a bacterial species is to be understood to include also its sub-species.

Throughout the specification, an ore is considered material that has been removed from the ground and does not receive any treatment to increase the metal concentration. A concentrate is produced by passing an ore through a treatment process, generally gravity or flotation, in order to increase the concentration of desired metals and decrease the volume of material which is subsequently treated to recover those desired metals.

DISCLOSURE OF THE INVENTION

In accordance with the present invention there is provided a bacterial culture for use in the bacterial oxidation of sulphide ores and concentrates, the bacterial culture identified by AGAL deposit Accession No. NM99/07541 or having been adapted therefrom, the bacterial culture containing least one or more strains of both Suifobaciilus anc Thermoplasma.

In accordance with the present invention there is further provided a process for the bacterial oxidation of sulphide ores and concentrates characterised in that the ore or concentrate is leached using either a bacterial culture identified by AGAL deposit Accession No. NM328/07541 or a bacterial culture adapted therefrom, the bacterial culture containing least one or more strains of both Sulfobacillus and

Thermoplasma.

: E -6- ir one form of the present invention the sulphide cre cr concentrate contains chaicopyrite.

The leach utilised in the process of the present invention may be conducted in a form selected from the group consisting of: a heap leach, a tank leach, a vat leach, and a dump leach.

The bacterial culture is preferably not indigenous to the ore or concentrate to be oxidised.

The bacterial culture and process of the present invention is effective in the oxidation of sulphide ores or concentrates when the ore or concentrate is provided at a grind or crush size of equal to or greater than Pgo 90pm. Preferably, the ore or concentrate is provided at a grind or crush size of equal to or greater than Pgo 75um.

In one form of the invention the culture is operable in the oxidation of sulphide ores and concentrates across a temperature range of 45 to 90°C. Preferably, the culture is operable in the oxidation of sulphide ores and concentrates across a temperature range of 45 to 65°C. .

BRIEF DESCRIPTION OF THE DRAWINGS :

The present invention will now be described, by way of example only, with reference to the accompanying Figure, in which:-

Figure 1 is a photographic representation of the denaturing gradient gel results for six samples of the bacterial culture of the present invention, - processed by three different methods.

DESCRIPTION :

In order fo raise a culture capable of processing chalcopyrite cres anc concentrates a bacterial culture indigenous to a chalcopyrite mineral was sought.

Indigenous bacterial cultures are typically superior to modified isolated cultures as the indigenous culture has already been adapted to the toxins and mineral components associated with a particular ore resulting in more effective and resilient bacterial strains.

Bacterial cultures indigenous to chalcopyrite ores were cultivated and tested for their ability to oxidise both their native ore/concentrate and other chalcopyrite ores and concentrates. During this program of work a stock bacteria culture was raised from a chalcopyrite (CuFeS) concentrate obtained from a base metal ore found in New Brunswick, Canada. Following the isolation of the bacterial culture, testing of the culture has taken place on both its’ native ore and concentrate, and on a variety of other ores and concentrates. Additions to the original stock culture have taken place as, during testing of the culture of different materials, any native bacteria capable of operating under the parameters of the test and being able to operate competitively with the introduced culture have not only survived but have thrived in the environment. In this way any bacteria native to the ore or concentrate being tested have been incorporated into the culture over time. In addition, the stock culture has been grown successfully at different temperatures ranging from 40° C to 90° C and at varying levels of acidity with pH levels ranging from 0.8-2.2. The applicants believe pH limits of 0.5 to 3.0 are not unreasonable.

Successful testing of the stock culture has taken place in both aerated agitated stirred tank reactors and in aerated columns to facilitate column leaching.

Successful testing of the stock culture has taken place at a variety of temperatures and on a variety of ores and concentrates.

The bacterial culture of the present invention consists of a variety of iron, sulphide and sulphur oxidising bacteria capable of working at temperatures of between 35°C and 90°C, and at pH ranges of between 0.5 and 3.0, although most preferably 0.8 and 2.5. The mixed bacterial culture is believed to include one or more representatives of both the genus’ Sulfebacillus and Thermoplasma,

notentially together with a number cf additionai bacteria! species. Such may 23 include, but is not limited to, Sulffobacillus thermosulfidooxidans, Thiobaciilis caldus and/or Thiobacillus ferrooxidans.

The mixed bacterial culture of the present invention has been deposited for the purpose of the Budapest Treaty at the Australian Government Analytical

Laboratories under Accession No. NM99/07541.

In the process for the bacterial oxidation of sulphide ores and concentrates of the present invention the stock bacterial culture described above is, in general terms, combined with the sample of the ore or concentrate to be leached as a process of adaptation. The stock culture is not indigenous to the ore or concentrate to which it is adapted. The adapted culture may then in turn be used to inoculate a heap of the ore or concentrate to which it has been adapted. Alternatively, other leach options may be used as a mechanism by which the adapted culture is exposed to the ore or concentrate, including vats, tanks and dumps.

The present invention will now be described by way of reference to a number of examples without limiting the generality of the invention described above.

Example 1

Prior to testing any material the stock bacterial culture, referred to hereinabove, is first adapted to the material of interest. This is facilitated by placing 2700 mi of modified OK solution (1.0 g/L ammonium sulphate, 0.5 g/L di-potassium orthophosphate, 0.16 g/L magnesium sulphate heptohydrate, pH 1.6 — 1.8) into an agitated aerated stirred tank reactor heated to the required temperature. To the modified 0K medium a 150g sample of milled (Pg<45 pm) test material is added and the pH adjusted down to: between 1.6 and 1.8 if necessary using concentrated sulphuric acid. To this slury a 300 mi slurry sample of the stock : inoculum is introduced. The agitated reactor is aerated at a rate of 1L/min/L slurry. By design, the stock inoculum is not indigenous to the ore of the test material. The adaption Is continued until the level of relevant metals reporting to solution reaches either 100% or reaches a plateau. Solution samples are assayed for metal ievels ir soiutior: through the use of an (CP, where sppreprizie the pH of the siurry is adjusted with concentrated sulphuric acid so that the or is between 1.6 and 1.8. in addition to metal ievels reporting to solution the progress : of the adaptionftest is further monitored according to its oxidation reduction potential (ORP), ferrous concentration and dissolved oxygen concentration (DO).

Once the culture has adapted to the material of interest it is used as an inoculum for further agitated aerated stirred tank reactor tests or as an inoculum for heap or column tests. The adapted bacterial inoculum is diluted further through the addition of an acidic basic nutrient solution containing ammonium sulphate, potassium orthophosphate and magnesium sulphate. The concentration of these nutrients in solution may vary between laboratory tests and commercial operation and between different commercial operations. In all cases the progress of oxidation is monitored through the levels of metals reporting to solution, pH, ORP, ferrous concentration and DO content.

The bacterial culture of the present invention was tested on a range of chalcopyrite bearing samples from various locations around the world. Table 1 below illustrates the mineralogy and origin of the chalcopyrite concentrates and ores tested using the bacterial culture of the present invention.

TABLE 1 | : [Sample [Mineralogy ~~ |Orgin 8 Molybdenum concentrate with low levels of copper in| Canada chalcopyrite.

C Concentrate comprising predominantly chalcopyrite (35%) Canada and cubanite (17%) with lesser quantities of pyrrhotite (10%) and minor amounts of pentiandite (3%) and sphalerite (3%).

D Copper Nickel concentrate in which copper is present as both USA racopyr (18.5 — 28.5%) and cubanite (15.8 — 30.8%). - | Nickel is present as pentlandite (17.7 — 10.4%) and occasionally replaced as violarite.

and minor amounts of bornite.

F Copper concentrate consisting of chalcocite (14%), | South 3 es chalcopyrite (10%), bornite (1%) and pyrite (1%). Africa ]

G Samples i and iii are ore samples and sample ii is a| Western concentrate sample. The sulphide minerals are | Australia predominantly pentlandite, chalcopyrite and pyrrhotite. ]

General test procedure

All tests on mineral samples were conducted in agitated aerated tank reactors. "Each test had a solids density of 10%w/v and was aerated by sparging at a rate of 1L air per minute per litre of slurry in the reactor. The evaporative losses due to the heating and aeration of the slurry were made up prior to sampling the tests.

This was accomplished through the addition of tap water. All slurries were made up in a proprietary nutrient media with a starting pH of 1.0. Sampling involved assaying the solution for iron, copper and other relevant metal ions. in addition, the oxidation-reduction potential (ORP), pH, ferrous iron and dissolved oxygen levels were also monitored and recorded. Copper release was used to monitor the progress of the test and once this reached a stable plateau or attained approximately 100% of the copper reporting to solution the test was deemed complete. Once complete the pulps were pressure filtered, the final leach liquor assayed and the filter cake washed with acidified water and dried. The dried filter cake was weighed and the residue assayed in order to conduct a metallurgical balance.

The results from head analysis, particle size analysis and the results following oxidation are summarised and displayed in Table 2.

TABLEZ

Sampie T°C| Days | Results

Leached after leachin

Particle Ea

Size % Leached

Analysis I

A Pg<90 um | 28.60 | 29.40 36 %.6 all :

I a alc lI I aL hh KA A Rl NL

Emi EE Rn

I Cc a NC Sl I . *nominal sizing of the “as received” concentrate.

A number of samples of the adapted bacterial culture of the present invention have been grown at temperatures ranging generally from 35° C to 65° C, although the inventors have noted operation at temperatures of up to about 90°C. Samples from each of the cultures have been removed and prepared for identification using 16SrRNA sequencing. Preparation of the samples prior to RNA sequencing was undertaken using three different methods. The methods used and the results obtained from 16SrRNA sequencing are as follows.

Methods . :

Six samples (designated SN45, SM45, PO45, SS 45, RH 14K, and 014A) were tested.

The sampies were mixed or: a hand shaker at maximum speed for 32 minties and processed as foliows:

A. Shaken. 500 pl of the shaken sample was immediately sedimented onto glass fibre filters (#30 Sliecher and Schuell, Keene, NH) in a 1.5 mi microtube by centrifugation at 14Krpm for 4 minutes. The supematant was carefully removed, and the sedimented material was washed twice in 1 mi of tissue culiure grade water.

B. Fast Prep. 500 ul was immediately removed and homogenized using a Savant

BIO 101 Fast Prep machine (BioCan Scientific) at speed 4 for 20 seconds. The homogenates were sedimented and washed as described above.

C. Supernatant. Following shaking, the samples were allowed to sit for 5 minutes to allow the particulate matter to settle to the bottom of the tubes. 500 pl of the supematant was then sedimented and washed as previously described.

RNA was extracted from ail samples using InstaGene Matrix (BioRad, Hercules,

CA) as per manufacture’s instructions. The RNA concentration was determined by uv spectrophotometry (Azo) and 50 ng were added to the PCR reaction mixture with a final concentration of 2mM magnesium ion, 100 uM dNTP, 0.32 pm each primer and 0.625 units of Taq Gold Polymerase. The universal primers p515f and p806r (Reiman 1993) were used to amplify an approximately 300bp segment of the 16S ribosomal RNA gene. The forward primer was modified with a 40bP GC rich sequence that terminated the migration of the ampiified product at various concentrations of urea/fcrmamide within a denaturing gradient gel (Sheffield et al. 1989; Muyzer et at. 1993) Bands of interest were cut from the "denaturing gels and purified amplified product was subjected to cycle sequencing using Big Dye Terminator extension from the reverse primer using the conditions recommended (PE Applied Biosystems). Sequence determination was performed on a 310 Genetic Analyser (PE Applied Biosystems). Sequence comparisons were conducted using the Basic Local Alignment Search Tool (BLAST; Altschul et al. 1990).

Results

Each of the three samples processing methods resulting in a different profile for the same sample, as shown in Figure 1. Nine predominant bands were selected for sequencing. The 300bp segments sequenced had the closest match with partial sequences of the 16S rRNA gene of the bacterial species listed in the

BLAST result column. A larger 16S segment would have to be sequenced for more precise identification.

A summary of the BLAST search results for the 300 base pair 16S rRNA gene segments sequenced is shown in Table 3. The numbers in parentheses refer to the % homology between the unknowns and their closest matches.

TABLE 3 band with same mobility BLAST result 1 SM45-fast prep SN45-shaken, C1 (1998) | Sulfobacillus shaken C/C (1998)-shaken | thermosulfidooxidans (98%) 2 C1 (1998)-shaken | SM45-fast prep SN45- | Sulfobacillus shaken CIC (1998)-shaken | thermosulfidooxidans (98%) 8 RH14K 60°C)- Unidentified ( ) PO45-fastprep/shaken/ entine supernatant supernatant : bacterium (97%)

SS45-supemnatant Denitrifying ~~ Fe<ll> oxidizing bacteria 197%) 014A 50°C Thiobacillus ( r SN45-fastprep/shaken/ supernatant supernatant ferrooxidans (96%) oo SM45-fastprep 014A shaken

It is envisaged that bacterial species may be omitted or substituted to the mixed culture outlined above in order to facilitate its operation at different temperatures.

For example, Thiobacillus thiooxidans a sulphur oxidising bacteria may be substituted fcr Thiobacillus caldus at lower temperatures.

The inventors considered that at least some of the initial results in the characterisation of the bacterial culture of the present invention may have been anomalous. Further work in the characterisation of the bacterial cuiture of the present invention has been undertaken as follows in Example 2. Over time the accuracy of such characterisation work increases due to improvements in search engines and the updating of databases.

Example 2

Methods

Denaturing gradient gel electrophoresis (“DGGE”) analysis of 16SrDNA fragments was performed on a sample of the bacterial culture of the present invention. This method, first described by Muyzer et al. (1993), is particularly suitable for profiling complex microbial populations.

Six aliquots of 10mL of the bacterial culture were centrifuged at ~13,000 g for 15 min. Total sample DNA was extracted from the pellets using a modified version of a previously described method (Plumb et al, 2001). Sample pellets were resuspended in pH 7.2 phosphate buffered saline to initiate cell lysis. Further cell lysis was obtained by treating samples with the lytic enzymes, lysozyme and proteinase K and also the strong detergent sodium dodecy! sulphate (SDS).

Zoliowing extraction of samples twice with phenol-chioroform-isoamy! aicohot,

DNA in solution was precipitated with isopropanol and sodium acetate. Extracted

DNA was further purified using the UltraClean™ PCR Clean-up Kit (MO BIO

Laboratories Inc). DNA samples were visualised by staining with ethidium bromide after electrophoresis through a 1% w/v agarose gel,

From each DNA sample, full-length 16S rRNA genes were amplified using the polymerase chain reaction (PCR). PCR primers specific for Bacteria and Archaea were used along with HotStarTaq polymerase (Qiagen) as described previously

{Plumb et al, 2002). PCR products were purified using the Ult-allean™ PTR . Clean-up, Kit and then used as template for DGGE PCR reactions. DGGE PCR was performed using previously described primer sets (Muyzer et al., 1993,

Ovreas et al, 1997). DNA from three reference strains, Leptospirillum ferroxidans, Sulfobacillus thermosulfidooxidans and Sulfolobus sp. Strain JP2 was also used to generate PCR fragments for DGGE analysis to provide a comparison. DGGE was performed using the DCode™ universal mutation detection system (BioRad Laboratories, USA), and 6% wiv polyacrylamide gels with a denaturing gradient ranging from 30% to 70% (where 100% denaturant : 10 contains 7 M urea and 40% v/v formamide). Electrophoreiss was conducted for 16 h at 100 V and 60°C. Gels were stained in 1XTAE buffer containing 0.5 mg L? ethidium bromide and documented using Multilmage™ light cabinet transilluminator TM-26 (Alpha Innotech Corporation, USA) and Chemilmage V5.5 software. Selected bands were excised from the gels and reamplified using PCR with DGGE primers. Purified PCR products were then sequenced using automated cycle sequencing as described previously (Plumb et al, 2002).

Sequence data was analysed using the Basic Local Alignment Search Tool (BLAST, Altschul et al., 1990) to compare sequences with sequence data ina non-redundant nucleic acid sequence database accessed through 2C hitp://www.ncbi.nim.nih.qov/BLAST/. :

Results

Examination of the sample using a phase contrast microscope revealed low numbers of small rod-shaped cells. DNA was successfully extracted from each of the six 10mL samples. The six samples were pooled during the DNA purification step resulting in three purified DNA samples.

From the purified genomic DNA, full-length 16S rDNA was amplified using

Bacteria-specific and Archaea-specific primers. This result showed that both

Bacteria and Archaea were present. These PCR products were purified and used : as templates for PCR amplification of DNA fragments for DGGE analysis. PCR using Bacteria-specific and Archaea-specific DGGE primers successfully amplified DNA fragments for DGGE analysis.

DCR ‘fragments were then subjected to analysis using DGGE tc separate INA fragments according tc their electrophoretic mobility through a get matrix containing increasing concentrations of denaturants. PCR samples from reference strains gave banding profiles. as expected. Analysis of the inoculum sample fragment generate using Bacteria-specific primers gave two feint, but distinctive bands. The sample fragment generated using Archaea-specific primers produced only one distinctive band on the gel. The rest of the profile showed non-distinct areas of smeared appearance. "PCR products from bands of interest were sequenced and analysed using BLAST to determine the identity of the DNA sequences. Results of BLAST analyses of the sequence data are summarised in Table 4. Band A contained DNA highly similar (99%) to DNA from strains of the genus Sulfobacillus. Bands B and GC contained DNA highly similar (98-99%) to DNA from unknown strains of

Thermoplasma. The genus Thermoplasma comprises organisms of the domain

Archaea that are characterised by their pleomorphic cell morphology due to the lack of a cell wall and their ability to grow at a range of temperatures from mesophilic to thermophilic. ‘Representatives of the genus are acidophiles capable of growing heterotrophically under aerobic and anaerobic conditions. : TABLE 4 . 20 BLAST analysis of sequence data from DNA bands excised from the DGGE profiles of the sample.

Band Label | Sample Information | Closest Match (% homology) (primers) - :

Inoculum (Bacteria) Sulfobacillus sp. G2 (99%)

B Inoculum (Baceria) Thermoplasma sp. clone ASL1 (99%)

Cc Inoculum (Archaea) Thermoplasma sp. clone ASL%

i is unexpected that a bacterial culture having the ability to oxidise suipitice cres and concentrates under the conditions described hereinabove would have representatives of both the genus’ Sulfobacillus and Thermoplasma, with potential for the addition of one or more of the bacterial species identified in Example 1.

It is envisaged that the materials the mixed bacterial culture of the present invention may be used to treat include base metal ores and concentrates (copper, nickel, cobalt zinc etc), precious metal ores and concentrates (gold and silver) and platinum group metal (PGM) ores and concentrates. It is further envisaged that the culture may be used in a heap leach, tank leach, vat leach or dump leach oxidation.

The bacterial culture and process of the present invention is capable of operating at a wide range of temperatures, thereby leading to a reduction in costs ~ associated with cooling bacterial oxidation systems. The process is further capable of oxidising all forms of chalcopyrite, and at crush sizes that need not 16 incur significant capital and operating costs.

Modifications and variations such as would be apparent to the skilled addressee are considered to fall within the scope of the present invention.

REFERENCES

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Brierley C.L. and R Brans, 1994 — Selection of BacTech’s Thermophilic Bio-

Oxidation Process for Youanmi Mine, In Biomine 94, Conference Proceeding

Perth Western Australia. Section 5.

Barrett, J., Hughes, M.N., Ewart, D.K. and Poole, RK, 1988 — The isolation and characterisation of a moderately thermophilic mixed culture of autotrophic bacteria: application: of the oxidation of refractory gold concentrates. Perth Goid 88, Randol International Ltd, Golden, Colorado, pp 148 150.

Dew, D.M. and D.M. Miler, 1997 — The BioNIC Process; Bioleaching of Mineral

Sulphide Concentrates For Recovery of Nickel, In IBS Biomine '97, Conference

Proceedings, Sydney, p M7.1.0 — M7.1.9.

Muyzer, G.E. C. DeWall, and A.G. Uitterlinden, 1993. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction amlified genes coding for the 16s rRNA. Appl. Environ.

Microbiol. 59: 695-700.

Nobar, A.M., Ewart, D.K., Alsaffar, L., Barrett, J., Hughes M.N. and Poole, RK, 1988 — Isolation and characterisation of a mixed microbial community from an

Australian mine: application to the leaching of gold from refractory ores,

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Letters, Kew Surrey, UK, pp 530-531.

Ovreds, L., Fomey, L., Daae, F.. and Torsvik, V. 1997. Distribution of bacterioplankton in meromictic Lake Saelenvannet, as determined by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA. Applied and Environmental Microbiology, 63, 3367-3373.

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Claims (13)

4 A } CLAIMS

1. A bacterial culture for use in the bacterial oxidation of sulphide ores and concentrates, the bacterial culture having been adapted from AGAL deposit Accession No. NM99/07541 and further containing one or more strains of both Sulfobacillus and Thermoplasma.

2. A process for the bacterial oxidation of sulphide ores and concentrates wherein the ore or concentrate is leached using a bacterial culture adapted from AGAL deposit Accession No. NM99/07541, the bacterial culture further containing one or more strains of both Sulfobacillus and Thermoplasma.

3. A process according to claim 2, wherein the sulphide ore or concentrate contains chalcopyrite.

4. A process according to claim 2 or 3, wherein the leach is conducted in a form selected from the group consisting of: a heap leach, a tank leach, a vat leach, and a dump leach.

5. A process according to any one of claims 2 to 4, wherein the bacterial culture is not indigenous to the ore or concentrate to be oxidised.

6. A process according to any one of claims 2 to 5, wherein the ore or concentrate is provided at a grind or crush size of equal to or greater than Pg 75um. AMENDED SHEET

JF AN ,

7. A process according tc any one of claims 2 tc §, wherein the cre or concentrate is provided at a grind or crush size of equal to or greater than Pgo 90pm.

8. A bacterial culture according to claim 1, wherein the culture is operable in the oxidation of sulphide ores and concentrates across a temperature range of 45 to 90°C.

9. A bacterial culture according to claim 1, wherein the culture is operable in the oxidation of sulphide ores and concentrates across a temperature range of 45 to 65°C.

10. A bacterial culture as claimed in any one of claims 1, 8 and 9, substantially as hereinbefore described or exemplified.

11. A bacterial culture according to the invention including any new and inventive integer or combination of integers, substantially as herein described.

12. A process according to the invention for the bacterial oxidation of sulphide ores and concentrates, substantially as hereinbefore described or exemplified.

13. A process for the bacterial oxidation of sulphide ores and concentrates including any new and inventive integer or combination of integers, substantially as herein described. AVMIENDED SHEET