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Thereafter, these samples were combined and then inoculated into a basal salts solution in which different substrates (ferrous sulfate, elemental sulfur, and chalcopyrite) served as energy sources. After that, the mixed cultures growing on different substrates were pooled equally, resulting in a final mixed culture. After being adapted to gradually increasing pulp densities of chalcopyrite concentrate by serial subculturing for more than 2 years, the final culture was able to efficiently leach the chalcopyrite at a pulp density of 20% (wt\/vol). At that pulp density, the culture extracted 60.4% of copper from the chalcopyrite in 25 days. The bacterial and archaeal diversities during adaptation were analyzed by denaturing gradient gel electrophoresis and constructing clone libraries of the 16S rRNA gene. The results show that the culture consisted mainly of four species, including\n                    <jats:named-content content-type=\"genus-species\">Leptospirillum ferriphilum<\/jats:named-content>\n                    ,\n                    <jats:named-content content-type=\"genus-species\">Acidithiobacillus caldus<\/jats:named-content>\n                    ,\n                    <jats:named-content content-type=\"genus-species\">Sulfobacillus acidophilus<\/jats:named-content>\n                    , and\n                    <jats:named-content content-type=\"genus-species\">Ferroplasma thermophilum<\/jats:named-content>\n                    , before adapting to a pulp density of 4%. However,\n                    <jats:named-content content-type=\"genus-species\">L. ferriphilum<\/jats:named-content>\n                    could not be detected when the pulp density was greater than 4%. Real-time quantitative PCR was employed to monitor the microbial dynamics during bioleaching at a pulp density of 20%. The results show that\n                    <jats:named-content content-type=\"genus-species\">A. caldus<\/jats:named-content>\n                    was the predominant species in the initial stage, while\n                    <jats:named-content content-type=\"genus-species\">S. acidophilus<\/jats:named-content>\n                    rather than\n                    <jats:named-content content-type=\"genus-species\">A. caldus<\/jats:named-content>\n                    became the predominant species in the middle stage.\n                    <jats:named-content content-type=\"genus-species\">F. thermophilum<\/jats:named-content>\n                    accounted for the greatest proportion in the final stage.\n                  <\/jats:p>","DOI":"10.1128\/aem.02907-13","type":"journal-article","created":{"date-parts":[[2013,11,16]],"date-time":"2013-11-16T00:55:13Z","timestamp":1384563313000},"page":"741-750","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":78,"title":["A Moderately Thermophilic Mixed Microbial Culture for Bioleaching of Chalcopyrite Concentrate at High Pulp Density"],"prefix":"10.1128","volume":"80","author":[{"given":"Yuguang","family":"Wang","sequence":"first","affiliation":[{"name":"School of Minerals Processing and Bioengineering, Central South University, Changsha, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weimin","family":"Zeng","sequence":"additional","affiliation":[{"name":"School of Minerals Processing and Bioengineering, Central South University, Changsha, China"},{"name":"Key Laboratory of Biometallurgy, Ministry of Education, Changsha, 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