Abstract
The microbial consortium involved in anaerobic digestion has not yet been precisely characterized and this process remains a ‘black box’ with limited efficiency. In this study, seven anaerobic sludge digesters were selected based on technology, type of sludge, process and water quality. The prokaryotic community of these digesters was examined by constructing and analysing a total of 9890 16S rRNA gene clones. Libraries were constructed using primers specific for the Bacteria and Archaea domains for each digester, respectively. After phylogenetic affiliation, the libraries were compared using statistical tools to determine the similarities or differences among the seven digesters. Results show that the prokaryotic community of an anaerobic digester is composed of phylotypes commonly found in all anaerobic digesters sampled and also of specific phylotypes. The Archaea community is represented by an equilibrium among a restricted number of operational taxonomic units (OTUs). These OTUs are affiliated with Methanosarcinales, Methanomicrobiales and Arc I phylogenetic groups. Statistical analysis revealed that the Bacteria community can be described as a three component model: one-third making up a core group of phylotypes common to most of the digesters, one-third are phylotypes shared among a few digesters and another one-third are specific phylotypes. The core group is composed of only six OTUs affiliated with Chloroflexi, Betaproteobacteria, Bacteroidetes and Synergistetes. Its role in anaerobic degradation appears critical to investigate. This comparison of anaerobic digester populations is a first step towards a future understanding of the relationship among biodiversity, operating conditions and digester efficiency.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ . (1990). Basic local alignment search tool. J Mol Biol 215: 403–410.
Ariesyady HD, Ito T, Okabe S . (2007). Functional bacterial and archaeal community structures of major trophic groups in a full-scale anaerobic sludge digester. Water Res 41: 1554–1568.
Ashelford KE, Chuzhanova NA, Fry JC, Jones AJ, Weightman AJ . (2005). At least 1 in 20 16S rRNA sequence records currently held in public repositories is estimated to contain substantial anomalies. Appl Environ Microbiol 71: 7724–7736.
Chao A . (1984). Non-parametric estimation of the number of classes in a population. Scand J Stat 11: 265–270.
Chao A, Yang MCK . (1993). Stopping rules and estimation for recapture debugging with unequal failure rates. Biometrika 80: 193–201.
Chouari R, Le Paslier D, Daegelen P, Ginestet P, Weissenbach J, Sghir A . (2003). Molecular evidence for novel planctomycete diversity in a municipal wastewater treatment plant. Appl Environ Microbiol 69: 7354–7363.
Chouari R, Le Paslier D, Daegelen P, Ginestet P, Weissenbach J, Sghir A . (2005a). Novel predominant archaeal and bacterial groups revealed by molecular analysis of an anaerobic sludge digester. Environ Microbiol 7: 1104–1115.
Chouari R, Le Paslier D, Dauga C, Daegelen P, Weissenbach J, Sghir A . (2005b). Novel major bacterial candidate division within a municipal anaerobic sludge digester. Appl Environ Microbiol 71: 2145–2153.
DeLong EF . (1992). Archaea in coastal marine environments. Proc Natl Acad Sci USA 89: 5685–5689.
DeSantis TZ, Hugenholtz P, Larsen N, Rojas M, Brodie EL, Keller K et al. (2006). Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Appl Environ Microbiol 72: 5069–5072.
Excoffier L, Smouse PE, Quattro JM . (1992). Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics 131: 479–491.
Felsenstein J, Churchill GA . (1996). A Hidden Markov Model approach to variation among sites in rate of evolution. Mol Biol Evol 13: 93–104.
Ficker M, Krastel K, Orlicky S, Edwards E . (1999). Molecular characterization of a toluene-degrading methanogenic consortium. Appl Environ Microbiol 65: 5576–5585.
Godon JJ, Moriniere J, Moletta M, Gaillac M, Bru V, Delgenes JP . (2005). Rarity associated with specific ecological niches in the bacterial world: the ‘Synergistes’ example. Environ Microbiol 7: 213–224.
Godon JJ, Zumstein E, Dabert P, Habouzit F, Moletta R . (1997a). Microbial 16S rDNA diversity in an anaerobic digester. Water Sci Technol 36: 49–55.
Godon JJ, Zumstein E, Dabert P, Habouzit F, Moletta R . (1997b). Molecular microbial diversity of an anaerobic digestor as determined by small-subunit rDNA sequence analysis. Appl Environ Microbiol 63: 2802–2813.
Good IJ . (1953). The population frequencies of species and the estimation of population parameters. Biometrika 40: 237–264.
Guermazi S, Daegelen P, Dauga C, Riviere D, Bouchez T, Godon JJ et al. (2008). Discovery and characterization of a new bacterial candidate division by an anaerobic sludge digester metagenomic approach. Environ Microbiol 10: 2111–2123.
Hicks RE, Amann RI, Stahl DA . (1992). Dual staining of natural bacterioplankton with 4′,6-diamidino-2-phenylindole and fluorescent oligonucleotide probes targeting kingdom-level 16S rRNA sequences. Appl Environ Microbiol 58: 2158–2163.
Howe K, Bateman A, Durbin R . (2002). QuickTree: building huge Neighbour-Joining trees of protein sequences. Bioinformatics 18: 1546–1547.
Juretschko S, Timmermann G, Schmid M, Schleifer KH, Pommerening-Roser A, Koops HP et al. (1998). Combined molecular and conventional analyses of nitrifying bacterium diversity in activated sludge: Nitrosococcus mobilis and Nitrospira-like bacteria as dominant populations. Appl Environ Microbiol 64: 3042–3051.
Katoh K, Misawa K, Kuma K, Miyata T . (2002). MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res 30: 3059–3066.
Kindaichi T, Ito T, Okabe S . (2004). Ecophysiological interaction between nitrifying bacteria and heterotrophic bacteria in autotrophic nitrifying biofilms as determined by microautoradiography-fluorescence in situ hybridization. Appl Environ Microbiol 70: 1641–1650.
Leclerc M, Delgènes J-P, Godon J-J . (2004). Diversity of the archael community in 44 anaerobic digesters as determined by single strand conformation polymorphism analysis and 16S rDNA sequencing. Environ Microbiol 6: 809–819.
Ley RE, Backhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI . (2005). Obesity alters gut microbial ecology. Proc Natl Acad Sci USA 102: 11070–11075.
Ludwig W, Strunk O, Westram R, Richter L, Meier H, Kumar Y et al. (2004). ARB: a software environment for sequence data. Nucleic Acids Res 32: 1363–1371.
Maidak BL, Larsen N, McCaughey MJ, Overbeek R, Olsen GJ, Fogel K et al. (1994). The Ribosomal Database Project. Nucleic Acids Res 22: 3485–3487.
Pelletier E, Kreimeyer A, Bocs S, Rouy Z, Gyapay G, Chouari R et al. (2008). ‘Candidatus cloacamonas acidaminovorans’: genome sequence reconstruction provides a first glimpse of a new bacterial division. J Bacteriol 190: 2572–2579.
Radajewski S, Ineson P, Parekh NR, Murrell JC . (2000). Stable-isotope probing as a tool in microbial ecology. Nature 403: 646–649.
Sakamoto M, Umeda M, Ishikawa I, Benno Y . (2000). Comparison of the oral bacterial flora in saliva from a healthy subject and two periodontitis patients by sequence analysis of 16S rDNA libraries. Microbiol Immunol 44: 643–652.
Schloss PD . (2008). Evaluating different approaches that test whether microbial communities have the same structure. ISME J 2: 265–275.
Schloss PD, Handelsman J . (2005). Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. Appl Environ Microbiol 71: 1501–1506.
Schloss PD, Handelsman J . (2006a). Introducing SONS, a tool for operational taxonomic unit-based comparisons of microbial community memberships and structures. Appl Environ Microbiol 72: 6773–6779.
Schloss PD, Handelsman J . (2006b). Introducing TreeClimber, a test to compare microbial community structures. Appl Environ Microbiol 72: 2379–2384.
Schloss PD, Larget BR, Handelsman J . (2004). Integration of microbial ecology and statistics: a test to compare gene libraries. Appl Environ Microbiol 70: 5485–5492.
Schlotelburg C, von Wintzingerode F, Hauck R, Hegemann W, Gobel UB . (2000). Bacteria of an anaerobic 1,2-dichloropropane-dechlorinating mixed culture are phylogenetically related to those of other anaerobic dechlorinating consortia. Int J Syst Evol Microbiol 50 (Pt 4): 1505–1511.
Sekiguchi Y, Kamagata Y . (2004). Strict and facultative anaerobes: medical and environmental aspects. In: Nakano MM, Zuber P (eds). Microbial Community Structure and Functions in Methane Fermentation Technology for Wastewater Treatment. Horizon Bioscience: UK, pp 361–384.
Sekiguchi Y, Kamagata Y, Syutsubo K, Ohashi A, Harada H, Nakamura K . (1998). Phylogenetic diversity of mesophilic and thermophilic granular sludges determined by 16S rRNA gene analysis. Microbiology 144: 2655–2665.
Sekiguchi Y, Takahashi H, Kamagata Y, Ohashi A, Harada H . (2001). In situ detection, isolation, and physiological properties of a thin filamentous microorganism abundant in methanogenic granular sludges: a novel isolate affiliated with a clone cluster, the green non-sulfur bacteria, subdivision I. Appl Environ Microbiol 67: 5740–5749.
Stach JEM, Maldonado LA, Masson DG, Ward AC, Goodfellow M, Bull AT . (2003). Statistical Approaches for Estimating Actinobacterial Diversity in Marine Sediments. Appl Environ Microbiol 69: 6189–6200.
Vartoukian SR, Palmer RM, Wade WG . (2007). The division ‘Synergistes’. Anaerobe 13: 99–106.
Zehnder AJB . (1988). Biology of Anaerobic Microorganisms. John Wiley & Sons, Inc.: New York. 872 p.
Zheng D, Alm EW, Stahl DA, Raskin L . (1996). Characterization of universal small-subunit rRNA hybridization probes for quantitative molecular microbial ecology studies. Appl Environ Microbiol 62: 4504–4513.
Acknowledgements
We are grateful to the Genoscope sequencing and informatics team and especially Simon Vallet for their technical support. We thank Sébastien Chaussonnerie, Isabelle Houdoin and Isabelle Bordelais for their technical assistance during the preparation of the libraries. We address a special acknowledgment to Denis Le Paslier for his advice and corrections on the article. We thank Dr Patrick Schloss for his help on SONS software. This study was supported by a grant from the ANRT.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on The ISME Journal website (http://www.nature.com/ismej)
Supplementary information
Rights and permissions
About this article
Cite this article
Rivière, D., Desvignes, V., Pelletier, E. et al. Towards the definition of a core of microorganisms involved in anaerobic digestion of sludge. ISME J 3, 700–714 (2009). https://doi.org/10.1038/ismej.2009.2
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/ismej.2009.2
Keywords
This article is cited by
-
MiDAS 5: Global diversity of bacteria and archaea in anaerobic digesters
Nature Communications (2024)
-
Functional and molecular approaches for studying and controlling microbial communities in anaerobic digestion of organic waste: a review
Reviews in Environmental Science and Bio/Technology (2023)
-
Studies on biodiversity and bioprospecting of active mud volcano associated bacteria and actinobacteria from Baratang, Andaman Islands, India
Systems Microbiology and Biomanufacturing (2023)
-
Effect of different multichannel ventilation methods on aerobic composting and vegetable waste gas emissions
Environmental Science and Pollution Research (2023)
-
Mechanism insights into enhanced treatment of wasted activated sludge by hydrogen-mediated anaerobic digestion
Environmental Science and Pollution Research (2023)


