Optimization of copper extraction for bioleaching of complex Cu-polymetallic concentrate by moderate thermophiles

Effects of initial pH, temperature, liquid volume, rotation speed, galvanic interaction （pyrite ratio） and pulp density on bioleaching of complex Cu-polymetallic concentrate were investigated. The results indicated that the copper extraction at pH 1.5 was 1.5 and 1.4 times that at pH 1.0 and pH 2.0 respectively. The copper extraction obtained at 45 ℃ was 1236.8%higher than that at 50 ℃. With the increase of rotation speed or the decrease of liquid volume, copper extraction was improved obviously. Copper extraction was improved gradually with the increase of pyrite ratio. However, when the ratio was higher than 20.0%, no further increase in copper extraction was observed. And the statistically significant interactive effects on copper extraction were found between temperature and pH, and temperature and pyrite ratio.

Comparative study on chalcopyrite bioleaching with assistance of different carbon materials by mixed moderate thermophiles

The catalysis of four carbon materials including artificial graphite(AG), carbon black(CB), activated carbon(AC) and carbon nanotube(CN) on chalcopyrite bioleaching by mixed moderate thermophiles was comparatively investigated. In AC and AG added bioleaching groups, low solution pH and suitable redox potential values, high total iron and ferric iron concentrations, and large number of adsorbed bacteria were obtained, resulting in high copper extractions. CB and CN inhibited the growth of bioleaching bacteria and led to the low bioleaching efficiency.X-ray diffraction analysis showed that jarosite and sulfur film were the main components of passivation layer with the addition of AG and AC,but did not hinder the dissolution of chalcopyrite. Microbial community structures of free and attached cells in AC and AG added groups changed dramatically compared with mixed moderate thermophiles. The sulfur-oxidizing bacteria of A. caldus S1 strain dominated the microbial community(93%-98%) at the end of bioleaching.The iron-oxidizing bacteria of L.ferriphilum YSK only accounted for low percentage(1%-2%).

Biodissolution of pyrite and bornite by moderate thermophiles

Acid mine drainage(AMD)has become a widespread environmental issue and its toxicity can cause permanent damage to the ecosystem.However,there are few studies focusing on the formation of AMD under moderately thermophilic conditions,hence we employed X-ray diffraction(XRD),scanning electron microscopy(SEM),X-ray photoelectron spectroscopy(XPS)and 16S rRNA sequencing to study the dissolution of pyrite and bornite by a moderate thermophilic consortium,and explored the role of free and attached microorganisms in the formation of AMD.The consortium mainly comprised Acidithiobacillus caldus,Leptospirillum ferriphilum and Sulfobacillus thermosulfidooxidans.The results indicated that total iron in pyrite solution system reached 33.45 g/L on the 12th day,and the copper dissolution rate of bornite dissolution reached 91.8%on the 24th day.SEM results indicated that the surfaces of pyrite and bornite were significantly corroded by microorganisms.XRD and XPS results showed that ore residues contained jarosite,and the dissolving residue of bornite contained elemental sulfur.The dominant bacterial genus in pyrite dissolution was A.caldus,and L.ferriphilum in bornite dissolution.To sum up,microbes significantly accelerated the mineral dissolution process and promoted the formation of AMD.

Split Addition of Nitrogen-Rich Substrate at Thermophilic and Mesophilic Stages of Composting: Effect on Green House Gases Emission and Quality of Compost

Composting as a solution to the increasing generation of municipal solid waste (MSW), also contribute to GHGs emission when not controlled and could lack some basic nutrients, especially nitrogen. This study assessed the split-additions of nitrogen-rich substrate to composting materials and their effect on GHGs emissions as well as the quality of the composts. Nitrogen-rich substrates formulated from pig and goat manure were co-composted with MSW for a 12-weeks period by split adding at mesophilic (˚C) and thermophilic (>50˚C) stages in five different treatments. Representative samples from the compost were taken from each treatment for physicochemical, heavy metals and bacteriological analysis. In-situ CH4, CO2, N2O gas emissions were also analyzed weekly during composting. It was observed that all the treatments showed significant organic matter decomposition, reaching thermophilic temperatures in the first week of composting. The absence affects the suitable agronomic properties. All nitrogen-rich substrate applied at thermophilic stage (Treatment two) recorded the highest N, P and K concentrations of 1.34%, 0.97% and 2.45%, respectively with highest nitrogen retention. In terms of GHG emissions, CO2 was highest at the thermophilic stage when N-rich substrate was added in all treatment, while CH4 was highest in the mesophilic stage with N-rich substrate addition. N2O showed no specific trend in the treatments. Split addition of the N-rich substrate for co-composting of MSW produced compost which is stable, has less nutrient loss and low GHG emissions. Split addition of a nitrogen-rich substrate could be an option for increasing the fertilizer value of MSW compost.

Characterization of apparent sulfur oxidation activity of thermophilic archaea in bioleaching of chalcopyrite

The apparent sulfur oxidation activities of four pure thermophilic archaea, Acidianus brierleyi （JCM 8954）, Metallosphaera sedula （YN 23）, Acidianus manzaensis （YN 25） and Sulfolobus metallicus （YN 24） and their mixture in bioleaching chalcopyrite were compared, which were characterized indirectly by the evolution of the cells concentration, pH value and sulfate ions concentration in solution. The results show that the mixed culture contributed significantly to the raising of leaching rate, which suggests that the mixed culture had a higher sulfur oxidation activity than the pure culture. Meanwhile, the results also indicate that the changes of parameters characterizing the sulfur oxidation activity of thermophilic archaea are often influenced by many factors, so it is hard to reflect accurately the specific sulfur oxidation activities among the different sulfur-oxidizing microbes when bioleaching chalcopyrite at different conditions. Accordingly, an efficient method to characterize microbial sulfur oxidation activity appears to be desirable.

Surface species of chalcopyrite during bioleaching by moderately thermophilic bacteria

X-ray diffraction （XRD） and X-ray photoelectron spectroscopy （XPS） analyses were carried out to investigate the surface species and interfacial reactions during bioleaching of chalcopyrite by different strains of moderately thermophilic bacteria （45 °C）. Results show that monosulfide （CuS）, disulfide （S22？）, polysulfide （Sn2？）, elemental sulfur （S0） and sulfate （SO42？） are the main intermediate species on the surface of chalcopyrite during bioleaching byA. caldus,S. thermosulfidooxidans andL. ferriphilum. The low kinetics of dissolution of chalcopyrite inA. caldus can be mainly attributed to the incomplete dissolution of chalcopyrite and the passivation layer of polysulfide. Polysulfide and jarosite should be mainly responsible for the passivation of chalcopyrite in bioleaching byL. ferriphilumorS. thermosulfidooxidans. However, elemental sulfur should not be the main composition of passivation layer of chalcopyrite during bioleaching.

Effect of moderately thermophilic bacteria on metal extraction and electrochemical characteristics for zinc smelting slag in bioleaching system

The effects of moderately thermophilic bacteria on the extraction of metals from zinc smelting slag and electrochemical characteristics of zinc smelting slag carbon paste electrode in bioleaching process were studied. The results show that the extraction rates of Fe, Cu and Zn from the slag reach 86.7%, 90.3% and 66.7% after adsorbed bacteria sterilize, while those with adsorbed bacteria are 91.9%, 96.0% and 84.5% in conditions of pulp density 2%, pH 1.0, temperature 65 °C and stirring rate 120 r/min, respectively. Some stretching peaks of functional groups from bacterial secretes on the bioleached residue surface, such as 1007 cm-1 and 1193 cm-1, turn up through FI-IR analysis and indirectly reveal the presence of the adsorbed bacteria on the slag particles surface. Besides, the corrosion of zinc smelting slag is enhanced by bacteria according to the characteristics of cyclic voltametry and Tafel curves in bioleaching system.

Feasibility study on heap bioleaching of chalcopyrite

Bioleaching of chalcopyrite often encountered the formation of passivation layer, which inhibited the leaching process and resulted in a low leaching rate. This inhibitory effect can be eliminated by thermophilic biole- aching. The industrial test of BioCOP technology based on thermophiles was successfully completed, which confirmed the feasibility of chalcopyrite bioleaching. However, industrial leaching rate of chalcopyrite heap bioleaching is lower. This paper described the development status and industrial test of chalcopyrite heap bioleaching technology. The reasons for the lower efficiency of chalcopyrite heap bioleaching were analyzed. The strategies for successful chalcopyrite heap bioleaching were proposed.

Improving zinc recovery by thermoacidophilic archaeon Acidianus copahuensis using tetrathionate

The attachment and bioleaching experiments were conducted to evaluate the zinc recovery from Hualilan ore by thethermoacidophilic archaeon Acidianus copahuensis. Cells of this species pregrown on tetrathionate showed higher capability ofattachment to the ore than cells pregrown on other energy sources and such attachment seemed to be mediated by the product ofextracellular polymeric substances. A. copahuensis achieved a successful bioleaching of the ore reaching 100% of zinc recoverywhen tetrathionate was added. Simultaneous addition of yeast extract and tetrathionate maintained the zinc extraction at higher rate.Zinc dissolution kinetics was controlled by chemical reaction in cultures with the external addition of tetrathionate but by thediffusion through a product layer of jarosite in the other cultures.

CHARACTERISTICS OF THERMOPHILIC ANAEROBIC ACIDOGENESIS OF STARCH WASTEWATER

Acidogenic dissimilation of synthetic starch wastewater (1 000～10 000 mg COD·L -1 ) was studied in a thermophilic (55 ℃) upflow anaerobic sludge blanket (UASB) reactor.The production of volatile fatty acids (VFA) was proportional to the chemical oxygen demand (COD) loading rate.The yield of VFA was around 0.28 g VFA/g COD over the COD loading rate from 1.25 to 30 g COD·L -1 ·d -1 and the hydraulic retention time from 8.8 h to 24 h.Distribution of organic acids,the contents of propionic and butyric acids in the effluent in particular were also dependent on the COD loading rate.The thermophilic UASB reactor showed a stable performance on hydrolysis and acidogenesis of starch as well as suspended solid removal at short hydraulic retention times and high influent pH(10～11),during the operation of 110 d.

Research on the Nature of Thermophilic Anaerobic Ethanol Producer Thermo anaerobacter sp DF3 in Petroleum Reservoirs

[Objective] The aim was to study the physiological and biochemical char- acteristics of Thermoanaerobacter sp DF3 in petroleum reservoirs and optimize the culture plan of producing ethanol from xylose. [Method] DF3, an anaerobic bacillus producing ethanol, was isolated from produced liquid from oil layer of Dagang oil field with anaerobic isolation technique. The phylogenetic position was analyzed by physiological and biochemical identification and phylogeny of 16S rDNA sequence. The metabolites were analyzed by gas chromatograph. [Result] The strain DF3 was a strict anaerobic thermophilic bacterium, which was straight in rod shape,and gram negative. Besides, it was 0.42 μmx（1.60-5.20） iJm in length. The strains can be soli- tary,in pairs or string and apical spore usually produced. Its growth temperature was 45-78 and 65 ℃ was the optimum. Many substances could be used as carbon sources, including glucose, xylose, fructose, ribose, mannose, arabinose, sucrose, galactose, lactose, cellobiose, melizitose, raffinose-, and starch. The similarity between strain DF3 and T. pseudoethanolicus achieved 99.7%. The main product of glucose and xylose fermentation was ethanol. After the culture plan was optimized,the final concentration of ethanol was 2.0 g/L. [Conclusion] It was proved through experiments that the strain DF3 was one of the strains with higher activity to produce ethanol at present and it could produce 2.0 g/L ethanol from xylose metabolization at 65 ℃. It has been demonstrated that DF3 is one of the known strains with high-production to produce ethanol,for example, 2.0 g/L ethano at 65℃. Presently, all the high-yield ethanol can be produced from metabolic xylose strains of metabolic xylose were isolat- ed by foreign countries, therefore, isolation of strain DF3 has provided an excellent original strain for studying ethanol production from lignocellulose in China.

深海嗜热菌超氧化物歧化酶基因耐盐性研究

为探究深海嗜热菌(Thermophile thermus)中铁离子超氧化物歧化酶Fe-SOD基因(Tt SOD)与植物耐盐性的关系,对该基因的全长序列进行了分析,构建了Tt SOD基因过量表达载体,并将该过量表达载体成功转化烟草中,获得转基因植株。结果表明,该基因全长615 bp,是典型的Fe-SOD。转Tt SOD基因烟草中超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性显著提高,而丙二醛(MDA)含量显著降低;转基因烟草的种子萌发率、叶绿素和类胡萝卜素含量显著提高。本研究的结果表明植物过表达Tt SOD基因,提高了转基因烟草耐盐能力,为选育作物耐盐性新品种提供技术参考。

Influence of Hot Spring Phages on Community Carbon Metabolism: Win, Lose or Draw?

Abundant virus-like particles were concentrated from large-volume samples from two hot springs. Both addition of viral concentrates and addition of samples induced by addition of mitomycin-C changed patterns of carbon source utilization by hot spring microbial communities. Specific effects of the two treatments depended upon both temperature and incubation period. Increased metabolic capability with greater exposure to free phages, consistent with the view that phages are major lateral transporters of metabolic genes, was observed most clearly in microbes incubated at a temperature lower than that encountered in situ. On the other hand, decreases in the diversity of utilizable C sources upon exposure to phages may have been due to lytic activity in which susceptible bacterial populations were differentially reduced by infective viruses, consistent with the “killing the winner” hypothesis. Treatment of cultures with MC-treated culture extracts, assumed to increase exposure to excised prophages, resulted in higher average metabolic rates after 18 h, but lower rates after 48 h of incubation. With incubation at in situ temperature, this same treatment led to an initial increase in the number of readily utilized C sources, followed by a decrease in community metabolic diversity relative to controls in samples from both hot springs. Thus, treatments designed to increase the interaction between hot spring microbes and either free or newly-excised phages had observable time- and temperature-dependent effects on community metabolism, demonstrating an important, yet complex, ecological role for phages in hot spring waters.

Start-up performances of dry anaerobic mesophilic and thermophilic digestions of organic solid wastes

Two dry anaerobic digestions of organic solid wastes were conducted for 6 weeks in a lab-scale batch experiment for investigating the start-up performances under mesophilic and thermophilic conditions. The enzymatic activities, i.e., β-glucosidase, N-α-benzoyl-Largininamide （BAA）-hydrolysing protease, urease and phosphatase activities were analysed. The BAA-hydrolysing protease activity during the first 2-3 weeks was low with low pH, but was enhanced later with the pH increase. β-Glucosidase activity showed the lowest values in weeks 1-2, and recovered with the increase of BAA-hydrolysing protease activity. Acetic acid dominated most of the total VFAs in thermophilic digestion, while propionate and butyrate dominated in mesophilic digestion. Thermophilic digestion was confirmed more feasible for achieving better performance against misbalance, especially during the start-up period in a dry anaerobic digestion process.

Autoheated thermophilic aerobic sludge digestion and metal bioleaching in a two-stage reactor system

A two-stage process has been developed for stabilization of sludge and removal of heavy metals from the secondary activated sludge with high rate of energy and time conservation. The first stage of the process involves autoheated thermophilic aerobic digestion at 55-60℃ inoculated with less-acidophilic thermophilic sulfur-oxidizing microorganisms （ATAD）. The results show that it is possible to maintain the autoheated conditions （55-60℃） in the ATAD reactor up to 24 hr, leading to reduction of 21% total solids （TS）, 27% volatile solids （VS）, 27% suspended solids （SS） and 33% volatile suspended solids （VSS） from the sludge. The sludge pH also decreased from 7 to 4.6 due to the activity of less-acidophilic thermophilic microorganisms. In the second stage operation, the digested sludge （pH 4.6, TS 31.6 g/L） from stage one was subjected to bioleaching in a continuous stirred tank reactor, operated at mean hydraulic retention times （HRTs） of 12, 24 and 36 hr at 30℃. An HRT of 24 hr was found to be sufficient for removal of 70% Cu, 70% Mn, 75% Ni, and 80% Zn from the sludge. In all, 39% VSS, 76% Cu, 78.2% Mn, 79.5% Ni and 84.2% Zn were removed from the sludge in both the stages.

Bioleaching of chalcopyrite by mixed culture of moderately thermophilic microorganisms

A mixed culture of moderately thermophilic microorganisms was enriched from acid mine drainages(AMDs)samples collected from several sulphide mines in China,and the bioleaching of chalcopyrite was conducted both in shake flask and bioreactor.The results show that in the shake flask,the mixture can tolerate 50 g/L chalcopyrite after being acclimated to gradually increased concentrations of chalcopyrite.The copper extraction increases obviously in bioleaching of chalcopyrite with moderately thermophilic microorganisms supplemented with 0.4 g/L yeast extract at 180 r/min,74% copper can be extracted in the pulp of 50 g/L chalcopyrite after 20 d.Compared with copper extractions of mesophilic culture,unacclimated culture and acclimated culture without addition of yeast extract,that of accliniated culture with addition of yeast extract is increased by 53%,44% and 16%,respectively.In a completely stirred tank reactor,the mass fraction of copper and total iron extraction reach up to 81% and 56%,respectively.The results also indicate that it is necessary to add a large amount of acid to the pulp to extract copper from chalcopyrite effectively.

Relatedness between catalytic effect of activated carbon and passivation phenomenon during chalcopyrite bioleaching by mixed thermophilic Archaea culture at 65°C

The relatedness between catalytic effect of activated carbon and passivation phenomenon during chalcopyrite bioleachingby mixed thermophilic Archaea culture(Acidianus brierleyi,Metallosphaera sedula,Acidianus manzaensis and Sulfolobusmetallicus)at65°C was studied.Leaching experiments showed that the addition of activated carbon could significantly promote thedissolution of chalcopyrite for both bioleaching and chemical leaching.The results of synchrotron-based X-ray diffraction,ironL-edge and sulfur K-edge X-ray absorption near edge structure spectroscopy indicated that activated carbon could change thetransition path of electrons through galvanic interactions to form more readily dissolved secondary mineral chalcocite at a low redoxpotential(?400mV)and then enhanced the copper dissolution.Jarosite accumulated immediately in the initial stage of bioleachingwith activated carbon but copper dissolution was not hindered.However,much jarosite precipitated on the surface of chalcopyrite inthe late stage of bioleaching,which might account for the decrease of copper dissolution rate.More elemental sulfur(S0)was alsodetected with additional activated carbon but the mixed thermophilic Archaea culture had a great sulfur oxidation activity,thus S0waseliminated and seemed to have no significant influence on the dissolution of chalcopyrite.

Comparison of leaching of bornite from different regions mediated by mixed moderately thermophilic bacteria

Bioleaching experiments combined with X-ray photoelectron spectroscopy(XPS),X-ray diffraction(XRD)and scanning electron microscopy(SEM)were conducted to investigate three kinds of bornites from different regions leached by moderately thermophilic mixed bacteria of Leptospirillum ferriphilum YSK,Acidithiobacillus caldus D1 and Sulfobacillus thermosulfidooxidans ST.The results of bioleaching experiments showed that the leaching efficiency and the redox potential were significantly increased.The copper extraction efficiencies of three kinds of bornite maintained rapid growth until around the 12th day and no longer increased after the 18th,reaching 83.7%,96.5%and 86.6%,respectively.The XRD results of the leaching residue indicated that three kinds of bornites all produced jarosite in the late stage of leaching,and the leaching residues from of Daye Museum and Yunnan Geological Museum contained a mass of elemental sulfur.XPS analysis and scanning electron microscopy experiments showed that the surface of mineral particles was jarosite and the copper in the leaching residue was almost dissolved.

Removal of copper from molybdenite concentrate by mesophilic and extreme thermophilic microorganisms

Mixed mesophilic and extreme thermophilic bioleaching were evaluated to remove copper from the molybdenite concentrate.Bioleaching tests were carried out in shake flasks and in a 50-L bioreactor.The shake flask tests were performed with different inoculum size,solids density,pH.and temperature in order to identify optimum conditions.The highest amount of copper elimination,75%was obtained with extreme thermophilic microorganisms(at 12%inoculation,10%solids,65℃and a pH of 1.5).The highest copper elimination by mesophilic microorganisms was 55%(at 12%inoculation,5%solids,30℃at pH 2).The optimum conditions in shake flask tests were applied to 7 days batch tests in a50-L bioreactor.Extreme thermophilic experiment gave the best copper elimination of 60%(at 12%inoculation,10%solids,65℃and pH 1.5).Mesophilic test removed 50%of the copper(at 12%inoculation,10%solids,35℃at pH 2).

Screening and Characterization of the Alkaline Protease Isolated from PLI-1, a Strain of Brevibacillus sp. Collected from Indonesia's Hot Springs

A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia's hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70℃ and pH 8.0-9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL-1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70℃ and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.