Bioreactor performance and microbial community dynamics in a production-scale biogas plant in northeastern China

Gao Yamei, Yang Anyi, Bao Jun, Ma Ruxia, Yan Lei, Wang Yanjie, Wang Weidong


In cold regions, heating is necessary to maintain the continuous and steady year-round operation of biogas fermentation. In this study, changes in the liquid composition, biogas production, and microbial diversity in heated- and unheated-phase samples were evaluated in a production-scale biogas plant that was fed continuously with cattle manure as a mono-substrate in Heilongjiang province in northeastern China. The volatile solid (VS) and volatile fatty acid (VFA) contents both gradually decreased in the heated and unheated fermentation processes. The chemical oxygen demand (COD) removal efficiency in the unheated phase sampled on June 15 (s-6-15) and October 15 (a-10-15) and in the heated phase sampled on January 15 (w-1-15) was 63.35%, 44.2% and 44.0%, respectively. The biogas production yields were in agreement with the results obtained for the VS and VFA contents and COD removal efficiency. The performance of the reactor in the heated phase was less efficient than that in the unheated phase, and the biogas production efficiency in June-August was higher than that in the other months. However, the CH4 content in the biogas remained similar all year. Moreover, ARDRA (Amplified Ribosomal DNA Restriction Analysis) was used to study the microbial community composition in the fermentation process. The results showed that the methanogenic archaeal consortium consisted mainly of members of the genera Methanomicrobiales and Methanosarcinales. In the heated phase, hydrogenotrophic methanogens represented the dominant methanogen in w-1-15 feedstock. After fermentation, the strict aceticlastic methanogen Methanosaeta became the dominant methanogen. In the unheated phase, the hydrogenotrophic methanogens and aceticlastic methanogens were equivalent in s-6-15 feedstock and effluent, and aceticlastic methanogens were dominant in both a-10-15 feedstock and effluent. Assessments of the bacteria diversity of the microbial communities revealed that the common strains in the feed and effluent of the three samples included the rumen bacteria, Bacteroides, Clostridium, Ruminococcaceae and Proteobacteria.
Keywords: biogas production, production-scale plant, dairy manure, microbial community, northeast of China
DOI: 10.3965/j.ijabe.20171001.2025

Citation: Gao Y M, Yang A Y, Bao J, Ma R X, Yan L, Wang Y J, et al. Bioreactor performance and microbial community dynamics in a production-scale biogas plant in northeastern China. Int J Agric & Biol Eng, 2017; 10(1): 191-201.


biogas production, production-scale plant, dairy manure, microbial community, northeast of China


Lin D. The development and prospective of bioenergy technology in China. Biomass and Bioenergy, 1998; 15(2): 181–186.

Naegele H J, Lindner J, Merkle W, Lemmer A, Jungbluth T, Bogenrieder C. Effects of temperature, pH and O2 on the removal of hydrogen sulfide from biogas by external biological desulfurization in a full scale fixed-bed trickling bioreactor (FBTB). Int J Agric & Biol Eng, 2013; 6(1): 69–81.

Zhang T, Mao C, Zhai N, Wang X, Yang G. Influence of initial pH on thermophilic anaerobic co-digestion of swine manure and maize stalk. Waste Manag., 2015; 35: 119–126.

Chang J, Leung D Y C, Wu C Z, Yuan Z H. A review on the energy production, consumption, and prospect of renewable energy in China. Renewable and Sustainable Energy Reviews, 2003; 7(5): 367–468.

Nansubuga I, Banadda N, Babu M, Vrieze J D, Verstraete W, Rabaey K. Enhancement of biogas potential of primary sludge by co-digestion with cow manure and brewery sludge. Int J Agric & Biol Eng, 2015; 8 (4):86–94.

Jiang X, Sommer S G, Christensen K V. A review of the biogas industry in China. Energy Policy, 2011; 39(10): 6073–6081.

Briones A, Raskin L. Diversity and dynamics of microbial communities in engineered environments and their implications for process stability. Curr. Opin. Biotechnol., 2003;14(3): 270–276.

Goberna M, Insam H, Klammer S, Pascual J A, Sánchez J. Microbial community structure at different depths in disturbed and undisturbed semiarid Mediterranean forest soils. Microb. Ecol., 2005; 50(3): 315–326.

Muyzer G, Smalla K. Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie Van Leeuwenhoek, 1998; 73(1): 127–141.

Wang W D, Yan L, Cui Z J, Gao Y M, Wang Y J, Jing R Y. Characterization of a microbial consortium capable of degrading lignocellulose. Bioresour. Technol., 2011; 102: 9321–9324.

Ventorino V, Aliberti A, Faraco V, Robertiello A, Giacobbe S, Ercolini D, et al. Exploring the microbiota dynamics related to vegetable biomasses degradation and study of lignocellulose-degrading bacteria for industrial biotechnological application. Sci. Rep., 2015; 5: 8161.

Sekiguchi Y, Kamagata Y, Syutsubo K, Ohashi A, Harada H, Nakamura K. Phylogenetic diversity of mesophilic and thermophilic granular sludges determined by 16S rRNA gene analysis. Microbiology, 1998; 144: 2655–2665.

Hori T, Haruta S, Ueno Y, Ishii M, Igarashi Y. Direct comparison of single-strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) to characterize a microbial community on the basis of 16S rRNA gene fragments. J. Microbiol. Methods, 2006; 66(1): 165–169.

Klocke M, Nettmann E, Bergmann I, Mundt K, Souidi K, Mumme J, et al. Characterization of the methanogenic Archaea within two-phase biogas reactor systems operated with plant biomass. Syst. Appl. Microbiol., 2008; 31(3): 190–205.

Goberna M, Gadermaier M, García C, Wett B, Insam H. Adaptation of methanogenic communities to the cofermentation of cattle excreta and olive mill wastes at 37°C and 55°C. Appl. Environ. Microbiol., 2010; 76(19): 6564–6571.

Ziganshin A M, Ziganshina E E, Kleinsteuber S, Pröter J, Ilinskaya O N. Methanogenic community dynamics during anaerobic utilization of agricultural Wastes. Acta Nature, 2012; 4(4): 91–97.

Karakashev D, Batstone D J, Angelidaki I. Influence of environmental conditions on methanogenic compositions in anaerobic biogas reactors. Appl. Environ. Microbiol., 2005; 71(1): 331–338.

Schlüter A, Bekel T, Diaz N N, Dondrup M, Eichenlaub R, Gartemann K H, et al. The metagenome of a biogas-producing microbial community of a production-scale biogas plant fermenter analysed by the 454-pyrosequencing technology. J. Biotechnol., 2008; 136(1-2): 77–90.

Kröber M, Bekel T, Diaz N N, Goesmann A, Jaenicke S, Krause L, et al. Phylogenetic characterization of a biogas plant microbial community integrating clone library 16S-rDNA sequences and metagenome sequence data obtained by 454-pyrosequencing. J. Biotechnol., 2009; 142(1): 38–49.

Nettmann E, Bergmann I, Pramschüfer S, Mundt K, Plogsties V, Herrmann C, et al. Polyphasic analyses of methanogenic archaeal communities in agricultural biogas plants. Appl. Environ. Microbiol., 2010; 76(8): 2540–2548.

Stolze Y, Zakrzewski M, Maus I, Eikmeyer F, Jaenicke S, Rottmann N, et al. Comparative metagenomics of biogas-producing microbial communities from production- scale biogas plants operating under wet or dry fermentation conditions. Biotechnol. Biofuels., 2015; 8: 14.

Zhou J Y, Li P F, Li G, Zhang Q G, Ding P, Wang S P, et al. Design and preliminary experimental research on a new biogas fermentation system by solar heat pipe heating. Int. J Agric. & Biol. Eng., 2016; 9(2):153–162.

APHA. Standard methods for the examination of water and wastewater. American Public Health Association, Washington. DC, USA, 2005; pp.150–187.

Zhu H, Qu F, Zhu L. Isolation of genomic DNAs from plants, fungi and bacteria using benzyl chloride. Nucleic Acids Res., 1993; 21: 5279–5280.

Wang Y X, Liu Q, Yan L, Gao Y M, Wang Y J, Wang W D. A novel lignin degradation bacterial consortium for efficient pulping. Bioresour. Technol., 2013; 139: 113–119.

Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol. Biol. Evol., 2007; 24(8): 1596–1599.

Lee D H, Lau A K, and Pinder K L. Development and performance of an alternative Biofilter System. J. Air Waste Manage. Assoc., 2001; 51: 78–85.

Hanreich A, Schimpf U, Zakrzewski M, Schlüter A, Benndorf D, Heyer R, et al. Metagenome and metaproteome analyses of microbial communities in mesophilic biogas-producing anaerobic batch fermentations indicate concerted plant carbohydrate degradation. Syst. Appl. Microbiol., 2013; 36(5): 330–338.

Harwood C S, Canale-Parola E. Ecology of spirochetes. Annu. Rev. Microbiol., 1984; 38: 161–192.

Klocke M, Mähnert P, Mundt K, Souidi K, Linke B. Microbial community analysis of a biogas-producing completely stirred tank reactor fed continuously with fodder beet silage as mono-substrate. Syst. Appl. Microbiol., 2007; 30(2): 139–151.

Koeck D E, Wibberg D, Maus I, Winkler A, Albersmeier A, Zverlov V V, et al. Complete genome sequence of the cellulolytic thermophile Ruminoclostridium cellulosi wild-type strain DG5 isolated from a thermophilic biogas plant. J. Biotechnol., 2014; 188: 136–137.

Yan L, Gao Y M, Wang Y J, Liu Q, Sun Z Y, Fu B R, et al. Diversity of a mesophilic lignocellulolytic microbial consortium which is useful for enhancement of biogas production. Bioresour. Technol., 2012; 11: 49–54.

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