Advertisement

Applied Microbiology and Biotechnology

, Volume 102, Issue 8, pp 3819–3830 | Cite as

Enriching ruminal polysaccharide-degrading consortia via co-inoculation with methanogenic sludge and microbial mechanisms of acidification across lignocellulose loading gradients

  • Yuying Deng
  • Zhenxing Huang
  • Wenquan Ruan
  • Hengfeng Miao
  • Wansheng Shi
  • Mingxing Zhao
Environmental biotechnology

Abstract

Using lignocellulosic materials as substrates, ruminal microbiota were co-inoculated with anaerobic sludge at different loading rates (LR) to study the microbial community in the semi-continuous mode. The results indicated that the highest CH4 yield reached 0.22 L/g volatile solid at LR of 4 g/L/day, which obtained 56–58% of the theoretical value. In the steady stage with LR of 2–4 g/L/day and slurry recirculation, copies of total archaea increased. Especially the Methanobacteriales increased significantly (p < 0.05) to 3.30 × 108 copies/mL. The microbial communities were examined by MiSeq 16S rRNA sequencing. Enriched hydrolytic bacteria mainly belonged to Clostridiales, including Ruminococcus, Ruminiclostridium, and Ruminofilibacter settled in the rumen. High-active cellulase and xylanase were excreted in the co-inoculated system. Acid-producing bacteria by fermentation were affiliated with Lachnospiraceae and Bacteroidales. The acidogen members were mainly Spirochaetaceae and Clostridiales. Syntrophic oxidation bacteria mainly consisted of Synergistetes, propionate oxidizers (Syntrophobacter and Pelotomaculum), and butyrate oxidizers (Syntrophus and Syntrophomonas). There had no volatile fatty acid (VFA) accumulation and the pH values varied between 6.94 and 7.35. At LR of 6 g/L/day and a recirculation ratio of 1:1, the hardly degradable components and total VFA concentrations obviously increased. The total archaea and Methanobacteriales then deceased significantly to 8.56 × 105 copies/mL and 4.14 × 103 copies/mL respectively (p < 0.05), which resulted in the inhibition of methanogenic activities. Subsequently, microbial diversity dropped, and the hydrolytic bacteria and syntrophic oxidizers obviously decreased. In contrast, the abundances of Bacteroidales increased significantly (p < 0.05). Acetate, propionate, and butyrate concentrations reached 2.02, 6.54, and 0.53 g/L, respectively, which indicated “acidification” in the anaerobic reactor. Our study illustrated that co-inoculated anaerobic sludge enriched the ruminal function consortia and hydrogenotrophic methanogens played an important role in anaerobic digestion of lignocelluloses.

Keywords

Co-inoculated consortia Lignocellulose digestion Anaerobic acidification 

Notes

Acknowledgments

The authors were grateful to the Enzyme Engineering Technology Center of Jiangsu for the help in the experiments of this work

Funding information

This work was funded by the National Natural Science Foundation of China [grant numbers 21506076, 51678279, and 51508230], National Science and Technological Support of China [grant number 2014BAC25B01], Fundamental Research Funds for the Central Universities [grant number JUSRP1703XNC], the fund of Jiangsu Key Laboratory of Anaerobic Biotechnology [grant number JKLAB201607], and the funds for green catalysis and applied enzyme team of Changzhou Vocational Institute of Engineering [111308002216006, 201713102015Y].

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_8877_MOESM1_ESM.pdf (827 kb)
ESM 1 (PDF 827 kb)

References

  1. APHA (2005) Standard methods for the examination of water and wastewater, 21th edn. American Public Health Association (APHA), New YorkGoogle Scholar
  2. Azman S, Khadem AF, van Lier JB, Zeeman G, Plugge CM (2015) Presence and role of anaerobic hydrolytic microbes in conversion of lignocellulosic biomass for biogas production. Crit Rev Env Sci Tech 45:2523–2564.  https://doi.org/10.1080/10643389.2015.1053727 CrossRefGoogle Scholar
  3. Dai X, Tian Y, Li J, Su X, Wang X, Zhao S, Liu L, Luo Y, Liu D, Zheng H, Wang J, Dong Z, Hu S, Huang L (2015) Metatranscriptomic analyses of plant cell wall polysaccharide degradation by microorganisms in the cow rumen. Appl Environ Microbiol 81:1375–1386.  https://doi.org/10.1128/AEM.03682-14 CrossRefPubMedPubMedCentralGoogle Scholar
  4. De Bok FAM, Harmsen HJM, Plugge CM, De Vries MC, Akkermans ADL, De Vos WM, Stams AJM (2005) The first true obligately syntrophic propionate-oxidizing bacterium, Pelotomaculum schinkii sp. nov., co-cultured with Methanospirillum hungatei, and emended description of the genus Pelotomaculum. Int J Syst Evol Micr 55:1697–1703.  https://doi.org/10.1099/ijs.0.02880-0 CrossRefGoogle Scholar
  5. Demirel B, Scherer P (2008) The roles of acetotrophic and hydrogenotrophic methanogens during anaerobic conversion of biomass to methane: a review. Rev Environ Sci Biotechnol 7:173–190.  https://doi.org/10.1007/s11157-008-9131-1 CrossRefGoogle Scholar
  6. Deng Y, Huang Z, Ruan W, Zhao M, Miao H, Ren H (2017a) Co-inoculation of cellulolytic rumen bacteria with methanogenic sludge to enhance methanogenesis of rice straw. Int Biodeterior Biodegrad 117:224–235.  https://doi.org/10.1016/j.ibiod.2017.01.017 CrossRefGoogle Scholar
  7. Deng Y, Huang Z, Zhao M, Ruan W, Miao H, Ren H (2017b) Effects of co-inoculating rice straw with ruminal microbiota and anaerobic sludge: digestion performance and spatial distribution of microbial communities. Appl Microbiol Biot 101:5937–5948.  https://doi.org/10.1007/s00253-017-8332-3 CrossRefGoogle Scholar
  8. FitzGerald JA, Allen E, Wall DM, Jackson SA, Murphy JD, Dobson AD (2015) Methanosarcina play an important role in anaerobic co-digestion of the seaweed Ulva lactuca: taxonomy and predicted metabolism of functional microbial communities. PLoS One 10:e0142603.  https://doi.org/10.1371/journal.pone.0142603 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Fondevila M, Dehority BA (1996) Interactions between Fibrobacter succinogenes, Prevotella ruminicola, and Ruminococcus flavefaciens in the digestion of cellulose from forages. J Anim Sci 74(3):678–684.  https://doi.org/10.2527/1996.743678x CrossRefPubMedGoogle Scholar
  10. Fuma R, Oyaizu S, Nukui Y, Ngwe T, Shinkai T, Koike S, Kobayashi Y (2012) Use of bean husk as an easily digestible fiber source for activating the fibrolytic rumen bacterium Fibrobacter succinogenes and rice straw digestion. Anim Sci J 83(10):696–703.  https://doi.org/10.1111/j.1740-0929.2012.01017.x CrossRefPubMedGoogle Scholar
  11. Hatamoto M, Kaneshige M, Nakamura A, Yamaguchi T (2014) Bacteroides luti sp. nov., an anaerobic, cellulolytic and xylanolytic bacterium isolated from methanogenic sludge. Int J Syst Evol Micr 64:1770–1774.  https://doi.org/10.1099/ijs.0.056630-0 CrossRefGoogle Scholar
  12. Hu Y, Shen F, Yuan H, Zou D, Pang Y, Liu Y, Zhu B, Chufo WA, Jaffar M, Li X (2014) Influence of recirculation of liquid fraction of the digestate (LFD) on maize stover anaerobic digestion. Biosyst Eng 127:189–196.  https://doi.org/10.1016/j.biosystemseng.2014.09.006 CrossRefGoogle Scholar
  13. Hu ZH, Yu HQ, Yue ZB, Harada H, Li YY (2007) Kinetic analysis of anaerobic digestion of cattail by rumen microbes in a modified UASB reactor. Biochem Eng J 37:219–225.  https://doi.org/10.1016/j.bej.2007.04.013 CrossRefGoogle Scholar
  14. Hu ZH, Yu HQ (2005) Application of rumen microorganisms for enhanced anaerobic fermentation of corn stover. Process Biochem 40(7):2371–2377.  https://doi.org/10.1016/j.procbio.2004.09.021 CrossRefGoogle Scholar
  15. Janssen PH, Kirs M (2008) Structure of the archaeal community of the rumen. Appl Environ Microbiol 74:3619–3625.  https://doi.org/10.1128/AEM.02812-07 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Karthikeyan OP, Visvanathan C (2013) Bio-energy recovery from high-solid organic substrates by dry anaerobic bio-conversion processes: a review. Rev Environ Sci Biotechnol 12:257–284.  https://doi.org/10.1007/s11157-012-9304-9 CrossRefGoogle Scholar
  17. Kröber M, Bekel T, Diaz NN, Goesmann A, Jaenicke S, Krause L, Miller D, Runte KJ, Viehöver P, Pühler A, Schlüter A (2009) Phylogenetic characterization of a biogas plant microbial community integrating clone library 16S-rDNA sequences and metagenome sequence data obtained by 454-pyrosequencing. J Biotechnol 142:38–49.  https://doi.org/10.1016/j.jbiotec.2009.02.010 CrossRefPubMedGoogle Scholar
  18. Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874.  https://doi.org/10.1093/molbev/msw054 CrossRefPubMedGoogle Scholar
  19. Lazuka A, Auer L, Bozonnet S, Morgavi DP, O'Donohue M, Hernandez-Raquet G (2015) Efficient anaerobic transformation of raw wheat straw by a robust cow rumen-derived microbial consortium. Bioresour Technol 196:241–249.  https://doi.org/10.1016/j.biortech.2015.07.084 CrossRefPubMedGoogle Scholar
  20. Lerm S, Kleyböcker A, Miethling-Graff R, Alawi M, Kasina M, Liebrich M, Würdemann H (2012) Archaeal community composition affects the function of anaerobic co-digesters in response to organic overload. Waste Manag 32:389–399.  https://doi.org/10.1016/j.wasman.2011.11.013 CrossRefPubMedGoogle Scholar
  21. Limam RD, Chouari R, Mazéas L, Wu TD, Li T, Grossin-debattista J, Guerquin-Kern JL, Saidi M, Landoulsi A, Sghir A, Bouchez T (2014) Members of the uncultured bacterial candidate division WWE1 are implicated in anaerobic digestion of cellulose. MicrobiologyOpen 3:157–167.  https://doi.org/10.1002/mbo3.144 CrossRefPubMedPubMedCentralGoogle Scholar
  22. Liu FH, Wang SB, Zhang JS, Zhang J, Yan X, Zhou HK, Zhao GP, Zhou ZH (2009) The structure of the bacterial and archaeal community in a biogas digester as revealed by denaturing gradient gel electrophoresis and 16S rDNA sequencing analysis. J Appl Microbiol 106:952–966.  https://doi.org/10.1111/j.1365-2672.2008.04064.x CrossRefPubMedGoogle Scholar
  23. Müller N, Worm P, Schink B, Stams AJ, Plugge CM (2010) Syntrophic butyrate and propionate oxidation processes: from genomes to reaction mechanisms. Env Microbiol Rep 2:489–499.  https://doi.org/10.1111/j.1758-2229.2010.00147.x CrossRefGoogle Scholar
  24. Miller T, Currenti E, Wolin M (2000) Anaerobic bioconversion of cellulose by Ruminococcus albus, Methanobrevibacter smithii, and Methanosarcina barkeri. Appl Microbiol Biot 54:494–498.  https://doi.org/10.1007/s002530000430 CrossRefGoogle Scholar
  25. Miura Y, Okabe S (2008) Quantification of cell specific uptake activity of microbial products by uncultured Chloroflexi by microautoradiography combined with fluorescence in situ hybridization. Environ Sci Technol 42:7380–7386.  https://doi.org/10.1021/es800566e CrossRefPubMedGoogle Scholar
  26. Morrison M, Pope PB, Denman SE, Mcsweeney CS (2009) Plant biomass degradation by gut microbiomes: more of the same or something new? Curr Opin Biotech 20:358–363.  https://doi.org/10.1016/j.copbio.2009.05.004 CrossRefPubMedGoogle Scholar
  27. Mussoline W, Esposito G, Giordano A, Lens P (2013) The anaerobic digestion of rice straw: a review. Crit Rev Env Sci Tec 43:895–915.  https://doi.org/10.1080/10643389.2011.627018 CrossRefGoogle Scholar
  28. Nelson MC, Morrison HG, Benjamino J, Grim SL, Graf J (2014) Analysis, optimization and verification of Illumina–generated 16S rRNA gene amplicon surveys. PLoS One 9:e94249–e94249.  https://doi.org/10.1371/journal.pone.0094249 CrossRefPubMedPubMedCentralGoogle Scholar
  29. Nordberg Å, Jarvis Å, Stenberg B, Mathisen B, Svensson BH (2007) Anaerobic digestion of alfalfa silage with recirculation of process liquid. Bioresour Technol 98:104–111.  https://doi.org/10.1016/j.biortech.2005.11.027 CrossRefPubMedGoogle Scholar
  30. Nyonyo T, Shinkai T, Mitsumori M (2014) Improved culturability of cellulolytic rumen bacteria and phylogenetic diversity of culturable cellulolytic and xylanolytic bacteria newly isolated from the bovine rumen. FEMS Microbiol Ecol 88:528–537.  https://doi.org/10.1111/1574-6941.12318 CrossRefPubMedGoogle Scholar
  31. Ozbayram EG, Kleinsteuber S, Nikolausz M, Ince B, Ince O (2017) Effect of bioaugmentation by cellulolytic bacteria enriched from sheep rumen on methane production from wheat straw. Anaerobe 46:122–130.  https://doi.org/10.1016/j.anaerobe.2017.03.013 CrossRefPubMedGoogle Scholar
  32. Piao H, Lachman M, Malfatti S, Sczyrba A, Knierim B, Auer M, Tringe SG, Mackie RI, Yeoman CJ, Hess M (2014) Temporal dynamics of fibrolytic and methanogenic rumen microorganisms during in situ incubation of switchgrass determined by 16S rRNA gene profiling. Front Microbiol 5:307.  https://doi.org/10.3389/fmicb.2014.00307 CrossRefPubMedPubMedCentralGoogle Scholar
  33. Romsaiyud A, Songkasiri W, Nopharatana A, Chaiprasert P (2009) Combination effect of pH and acetate on enzymatic cellulose hydrolysis. J Environ Sci 21:965–970.  https://doi.org/10.1016/S1001-0742(08)62369-4 CrossRefGoogle Scholar
  34. Rychlik JL, May T (2000) The effect of a methanogen, Methanobrevibacter smithii, on the growth rate, organic acid production, and specific ATP activity of three predominant ruminal cellulolytic bacteria. Curr Microbiol 40:176–180.  https://doi.org/10.1007/s002849910035 CrossRefPubMedGoogle Scholar
  35. Shakeri Yekta S, Gonsior M, Schmitt-Kopplin P, Svensson BH (2012) Characterization of dissolved organic matter in full scale continuous stirred tank biogas reactors using ultrahigh resolution mass spectrometry: a qualitative overview. Environ Sci Technol 46:12711–12719.  https://doi.org/10.1021/es3024447 CrossRefGoogle Scholar
  36. Steinmetz RLR, Mezzari MP, da Silva MLB, Kunz A, do Amaral AC, Tápparo DC, Soares HM (2016) Enrichment and acclimation of an anaerobic mesophilic microorganism’s inoculum for standardization of BMP assays. Bioresour Technol 219:21–28.  https://doi.org/10.1016/j.biortech.2016.07.031 CrossRefPubMedGoogle Scholar
  37. Sun L, Liu T, Müller B, Schnürer A (2016) The microbial community structure in industrial biogas plants influences the degradation rate of straw and cellulose in batch tests. Biotechnol Biofuels 9:128.  https://doi.org/10.1186/s13068-016-0543-9 CrossRefPubMedPubMedCentralGoogle Scholar
  38. Van Soest PJ, Robertson JB, Lewis BA (1991) Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci 74:3583–3597.  https://doi.org/10.3168/jds.S0022-0302(91)78551-2 CrossRefPubMedGoogle Scholar
  39. Wang H, Vuorela M, Keränen AL, Lehtinen TM, Lensu A, Lehtomäki A, Rintala J (2010) Development of microbial populations in the anaerobic hydrolysis of grass silage for methane production. FEMS Microbiol Ecol 72:496–506.  https://doi.org/10.1111/j.1574-6941.2010.00850.x CrossRefPubMedGoogle Scholar
  40. Xiao KK, Guo CH, Zhou Y, Maspolim Y, Wang JY, Ng WJ (2013) Acetic acid inhibition on methanogens in a two-phase anaerobic process. Biochem Eng J 75:1–7.  https://doi.org/10.1016/j.bej.2013.03.011 CrossRefGoogle Scholar
  41. Zhang H, Zhang P, Ye J, Wu Y, Fang W, Gou X, Zeng G (2016) Improvement of methane production from rice straw with rumen fluid pretreatment: a feasibility study. Int Biodeterior Biodegrad 113:9–16.  https://doi.org/10.1016/j.ibiod.2016.03.022 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Changzhou Vocational Institute of EngineeringChangzhouChina
  2. 2.School of Environmental and Civil EngineeringJiangnan UniversityWuxiChina
  3. 3.Jiangsu Key Laboratory of Anaerobic BiotechnologyWuxiChina
  4. 4.Jiangsu Collaborative Innovation Center of Technology and Material of Water TreatmentSuzhouChina

Personalised recommendations