Characterization of microbial community dynamics during the anaerobic co-digestion of thermally pre-treated slaughterhouse wastes with glycerin addition

  • Á. Rodríguez-AbaldeEmail author
  • M. Guivernau
  • F. X. Prenafeta-Boldú
  • X. Flotats
  • B. Fernández
Research Paper


Microbial community dynamics during the anaerobic co-digestion of pig manure, pasteurized slaughterhouse waste and glycerin were studied in a lab-scale CSTR. The feed composition was optimized through progressive co-substrate additions for enhanced methane production and organic matter removal without accumulation of intermediate compounds. Microbial community structure of biomass samples was studied by means of qPCR and DGGE profiling of 16S rRNA genes (Bacteria and Archaea), and genus-specific qPCR of the methyl coenzyme M reductase gene (mcrA), which encodes for an enzyme universally involved in methanogenesis. The composition of the dominant bacterial populations remained relatively stable, when compared to those in the influent, but the highest changes were observed upon the introduction of glycerin. Biodiversity of archaea was restricted to a few representatives of the genera Methanosaeta and Methanosarcina, but Methanospirillum sp. was detected only when glycerin was introduced in the feeding. Glycerin supplementation coincided with the strongest increase in methane yield (from 0.22 to 0.64 m3CH4 m−3 d−1).


Co-digestion Microbial community Slaughterhouse waste 16S rDNA DGGE profiling qPCR of mcrA functional genes 



Animal by-products


Total chemical oxygen demand


Continuous stirred tank reactor


Denaturing gradient gel electrophoresis


Hydraulic retention time


Methyl-coenzyme M reductase






Chain fatty acids


Organic loading rate


Polymerase chain reaction


Pig manure


Pasteurized animal by-product


Quantitative time PCR


Residual glycerin


Ribosomal ribonucleic acid


Total alkalinity


Total solids


Volatile fatty acids


Volatile solids



This work was supported by the Spanish Ministry of Science and Innovation (project PROBIOGAS PSE-120000–2008-57v). The support of the CERCA Program and of the Consolidated Research Group TERRA (ref. 2017 SGR 1290), both from the Generalitat de Catalunya, is also acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Á. Rodríguez-Abalde
    • 1
    • 2
    Email author
  • M. Guivernau
    • 2
  • F. X. Prenafeta-Boldú
    • 2
  • X. Flotats
    • 3
  • B. Fernández
    • 2
  1. 1.EnergyLabVigoSpain
  2. 2.GIRO ProgramInstitute of Agrifood Research and Technology (IRTA)BarcelonaSpain
  3. 3.GIRO Joint Research Unit IRTA-UPC, Department of Agrifood Engineering and BiotechnologyUniversitat Politècnica de Catalunya (UPC)BarcelonaSpain

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