Bacterial Community Dynamics During the Application of a Myxococcus xanthus-Inoculated Culture Medium Used for Consolidation of Ornamental Limestone |
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Authors: | Guadalupe Piñar Concepcion Jimenez-Lopez Katja Sterflinger Jörg Ettenauer Fadwa Jroundi Antonia Fernandez-Vivas Maria Teresa Gonzalez-Muñoz |
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Institution: | (1) Institute of Applied Microbiology, Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Muthgasse 18, 1190 Vienna, Austria;(2) Department of Microbiology, University of Granada, Fuentenueva s/n, 18071 Granada, Spain |
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Abstract: | In this study, we investigated under laboratory conditions the bacterial communities inhabiting quarry and decayed ornamental
carbonate stones before and after the application of a Myxococcus xanthus-inoculated culture medium used for consolidation of the stones. The dynamics of the community structure and the prevalence
of the inoculated bacterium, M. xanthus, were monitored during the time course of the consolidation treatment (30 days). For this purpose, we selected a molecular
strategy combining fingerprinting by denaturing gradient gel electrophoresis (DGGE) with the screening of eubacterial 16S
rDNA clone libraries by DGGE and sequencing. Quantification of the inoculated strain was performed by quantitative real-time
PCR (qPCR) using M. xanthus-specific primers designed in this work. Results derived from DGGE and sequencing analysis showed that, irrespective of the
origin of the stone, the same carbonatogenic microorganisms were activated by the application of a M. xanthus culture. Those microorganisms were Pseudomonas sp., Bacillus sp., and Brevibacillus sp. The monitoring of M. xanthus in the culture media of treated stones during the time course experiment showed disparate results depending on the applied
technique. By culture-dependent methods, the detection of this bacterium was only possible in the first day of the treatment,
showing the limitation of these conventional techniques. By PCR-DGGE analysis, M. xanthus was detected during the first 3–6 days of the experiment. At this time, the population of this bacterium in the culture media
varied between 108–106 cells ml−1, as showed by qPCR analyses. Thereafter, DGGE analyses showed to be not suitable for the detection of M. xanthus in a mixed culture. Nevertheless, qPCR analysis using specific primers for M. xanthus showed to be a more sensitive technique for the detection of this bacterium, revealing a population of 104 cells ml−1 in the culture media of both treated stones at the end of the consolidation treatment. The molecular strategy used in this
study is proposed as an effective monitoring system to evaluate the impact of the application of a bacterially induced carbonate
mineralization as restoration/conservation treatment for ornamental stones. |
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