Detection of Methanogens in Anaerobic Granular Sludge by FISH Targeting for Functional Genes

Abstract:

Methanogens play an important role in the anaerobic digestion and production of methane, and show significant influence on the performance of anaerobic wastewater treatment process. Then the methanogens in anaerobic granular sludge samples from full-scale UASB bioreactors treating avermectin or starch wastewater were detected by FISH using 16S rRNA gene-based probe and functional gene-based probes. The results showed that the hybridization of methanogens in simultaneous FISH with mcrA gene-based and 16S rRNA gene-based probes was high coincident and the coincidence degree was about 60%-80%, implying the preferable hybridization consistency between functional gene-based probe and 16S rRNA gene-based probe. The relative abundance of methanogens obtained in FISH analysis using 16S rRNA gene-based probe seemed higher than that using functional gene-based probes, indicating that functional gene could provide more specific detection of methanogens than 16S rRNA gene probably. For functional gene-based probes, the methanogen-specificity was in the following order: mcrA＜F420＜mtr. During the development process of both granular sludge samples, the maximum relative abundance of methanogens was obtained at its mature phase using all probes. The relative abundance of methanogens in the sludge sample treating avermectin wastewater was lower than that treating starch wastewater, indicating smaller methanogenic population in the sludge sample treating avermectin wastewater due to inhibitory effect of antibiotic residue on methanogens probably.

• Periodical: Advanced Materials Research (Volumes 518-523)
• Pages: 299-304
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.518-523.299
• Online since: May 2012
• Authors: Run Zhang, Chun Liu, Liang Li, Hui Na Yang, Jing Liang Yang
• Keywords: Anaerobic Granular Sludge, Fish, Functional Gene, Methanogens

References

[1] L.W. Hulshoff Pol, S.I. de Castro Lopes, G. Lettinga and P.N.L. Lens: Water Res. Vol. 38 (2004), pp.1376-1389

DOI: 10.1016/j.watres.2003.12.002

[2] A.T. Akarsubasi, O. Ince, A.N. Oz, B. Kırdar and B.K. Ince: Process Biochem. Vol. 41 (2006), pp.28-35

DOI: 10.1016/j.procbio.2005.01.029

[3] A. Vlyssides, E.M. Barampouti and S. Mai: J. Environ. Manage. Vol. 86 (2008), pp.660-664

[4] J. O'Reilly, C. Lee, G. Collins, F. Chinalia, T. Mahony and V. O'Flaherty: Water Res. Vol. 43 (2009), pp.3365-3374

[5] J.L. Sanz and T. Köchling: Process Biochem. Vol. 42 (2007), pp.119-133

[6] J.R. Coleman, D.E. Culley, W.B. Chrisler and F.J. Brockman: J. Microbiol. Methods Vol. 71 (2007), pp.246-255

[7] Xian Zhang, A. Tani, F. Kawai and K. Kimbara: J. Biosci. Bioeng. Vol. 110 (2010), pp.716-719

[8] T. Tagawa, Y. Sekiguchil, A. Ohashi, H. Harada and K. Syutsubo: Water Sci. Technol. Vol. 42 (2000), pp.77-82

[9] G. Crocetti, M. Murto and L. Björnsson: J. Microbiol. Methods Vol. 65 (2006), pp.194-201

[10] K. Kubota, A. Ohashi, H. Imachi and H. Harada: J. Microbiol. Methods Vol. 66 (2006), pp.521-528

[11] G. Rastogi, D.R. Ranade, T.Y. Yeole, M.S. Patole and Y.S. Shouche: Bioresource Technol. Vol. 99 (2008), pp.5317-5326

DOI: 10.1016/j.biortech.2007.11.024

[12] W.T. Liu, O.C. Chan and H.H.P. Fang: Water Res. Vol. 36 (2002), pp.1767-1775

[13] W. Jupraputtasri, N. Boonapatcharoen and S. Cheevadhanarak: J. Microbiol. Methods Vol. 61 (2005), pp.95-104

[14] M.W. Friedrich: Methyl-coenzyme M Reductase: Methods Enzymol. Vol. 397 (2005), pp.428-442

[15] T. Lienard and G. Gottschalk: FEBS Letters Vol.425 (1998), pp.204-208

[16] M. Song, S.G. Shin and S.H. Wang: Bioresource Technol. Vol. 101 (2010), pp.523-528

[17] W. Jupraputtasri, N. Boonapatcharoen, S. Cheevadhanarak, P. Chaiprasert, M. Tanticharoen and S. Techkarnjanaruk: J. Microbiol. Methods Vol.61 (2005), pp.95-104

DOI: 10.1016/j.mimet.2004.11.017