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Quantification of methanogens by fluorescence in situ hybridization with oligonucleotide probe

  • Environmental Biotechnology
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Abstract

To monitor anaerobic environmental engineering system, new method of quantification for methanogens was tested. It is based on the measurement of specific binding (hybridization) of 16S rRNA-targeted oligonucleotide probe Arc915, performed by fluorescence in situ hybridization (FISH) and quantified by fluorescence spectrometry. Average specific binding of Arc915 probe was 13.4±0.5 amol/cell of autofluorescent methanogens. It was 14.3, 13.3, and 12.9 amol/cell at the log phase, at stationary phase and at the period of cell lysis of batch culture, respectively. Specific binding of Arc915 probe per 1 ml of microbial sludge suspension from anaerobic digester linearly correlated with concentration of autofluorescent cells of methanogens. Coefficient of correlation was 0.95. Specific binding of oligonucleotide probe Arc915 can be used for the comparative estimation of methanogens during anaerobic digestion of organic waste. Specific binding of Arc915 probe was linear function of anaerobic sludge concentration when it was between 1.4 and 14.0 mg/ml. Accuracy of the measurements in this region was from 5 to 12%.

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Acknowledgements

The research was supported by the National Environment Agency, the Ministry of the Environment, Singapore, and by Nanyang Technological University, Singapore.

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  1. School of Civil and Environmental Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798, Singapore

    O. Stabnikova, X.-Y. Liu, J.-Y. Wang & V. Ivanov

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  1. O. Stabnikova
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  2. X.-Y. Liu
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  3. J.-Y. Wang
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  4. V. Ivanov
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Correspondence to O. Stabnikova.

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Stabnikova, O., Liu, XY., Wang, JY. et al. Quantification of methanogens by fluorescence in situ hybridization with oligonucleotide probe. Appl Microbiol Biotechnol 73, 696–702 (2006). https://doi.org/10.1007/s00253-006-0490-7

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  • DOI: https://doi.org/10.1007/s00253-006-0490-7

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