Quantitative Monitoring of Microbial Species during Bioleaching of a Copper Concentrate
Quantitative Monitoring of Microbial Species during Bioleaching of a Copper Concentrate
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DOI：
10.3339/fmicb.2016.02044
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发表时间：
2016-12-20
影响因子：
5.2
通讯作者：
Joulian, Catherine
中科院分区：
文献类型：
--
作者：
Hedrich, Sabrina;Guezennec, Anne-Gwenaelle;Joulian, Catherine
对浸出过程中的微生物群落进行监测是控制浸出过程参数和提高浸出效率的必要条件。然而，合适的方法是有限的，因为它们通常不适合生物浸出样品，并且在这些类型的财团的文献中通常没有分类群特异性测定。因此，我们的研究重点是开发新的定量实时荧光定量PCR （qPCR）方法来定量酸性钙化硫杆菌。嗜铁钩端螺旋体、氧化硫硫杆菌和有益硫杆菌，并将结果与其他常用分子监测方法的数据进行比较，以评价其准确性和特异性。用于铜精矿浸出的搅拌槽生物反应器，容纳了一个与几个生物浸出操作相关的嗜酸、适度嗜热细菌联合体，作为模型系统。通过qPCR分析微生物群落，可以精确监测总生物量的演变以及特定物种的丰度。同一样品的标准指纹图谱、末端限制性片段长度多态性（T-RFLP）和毛细管电泳单链构象多态性（CESSCP）数据与qPCR数据趋势一致。然而，qPCR的主要附加价值是为每个物种提供定量数据，而从T-RFLP和CE-SSCP谱只能推断出相对丰度。通过应用另外两种不需要核酸提取的定量方法，SYBR Green染色后的总细胞计数和通过微量热法测量金属硫化物氧化活性，获得了额外的价值。总的来说，这些互补的方法允许在各种生物浸出操作中进行有效的定量微生物群落监测。
Monitoring of the microbial community in bioleaching processes is essential in order to control process parameters and enhance the leaching efficiency. Suitable methods are, however, limited as they are usually not adapted to bioleaching samples and often no taxon-specific assays are available in the literature for these types of consortia. Therefore, our study focused on the development of novel quantitative real-time PCR (qPCR) assays for the quantification of Acidithiobacillus caldus. Leptospirillum ferriphilurn, Sulfobacillus the rmosulfidooxidans, and Sulfobacillus benefaciens and comparison of the results with data from other common molecular monitoring methods in order to evaluate their accuracy and specificity. Stirred tank bioreactors for the leaching of copper concentrate, housing a consortium of acidophilic, moderately thermophilic bacteria, relevant in several bioleaching operations, served as a model system. The microbial community analysis via qPCR allowed a precise monitoring of the evolution of total biomass as well as abundance of specific species. Data achieved by the standard fingerprinting methods, terminal restriction fragment length polymorphism (T-RFLP) and capillary electrophoresis single strand conformation polymorphism (CESSCP) on the same samples followed the same trend as qPCR data. The main added value of qPCR was, however, to provide quantitative data for each species whereas only relative abundance could be deduced from T-RFLP and CE-SSCP profiles. Additional value was obtained by applying two further quantitative methods which do not require nucleic acid extraction, total cell counting after SYBR Green staining and metal sulfide oxidation activity measurements via microcalorimetry. Overall, these complementary methods allow for an efficient quantitative microbial community monitoring in various bioleaching operations.