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Frontiers of Environmental Science & Engineering

ISSN 2095-2201

ISSN 2095-221X(Online)

CN 10-1013/X

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Front. Environ. Sci. Eng.    2018, Vol. 12 Issue (4) : 10    https://doi.org/10.1007/s11783-018-1067-2
RESEARCH ARTICLE
Methanogenic community structure in simultaneous methanogenesis and denitrification granular sludge
Yujiao Sun1, Juanjuan Zhao1,3(), Lili Chen2, Yueqiao Liu1, Jiane Zuo2()
1. College of Water Science, Beijing Normal University, Beijing 100875, China
2. School of Environment, Tsinghua University, Beijing 100084, China
3. Environmental Engineering Department, North China Institute of Science and Technology, Beijing 101601, China
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Abstract

• UASB with SMD granules was operated with high removal efficiency of COD and NO3--N.

• Methanosaetaceae was absolute predominant methanogen in SMD granules.

• The methanogen quantity and activity decreased as C/N decreased from 20:1 to 5:1.

• Bacterial community succession happened with C/N decreasing.

A laboratory scale up-flow anaerobic sludge bed (UASB) bioreactor fed with synthetic wastewater was operated with simultaneous methanogenesis and denitrification (SMD) granules for 235 days with a gradient decrease of C/N. Molecular cloning, qRT-PCR and T-RFLP were applied to study the methanogenic community structures in SMD granules and their changes in response to changing influent C/N. The results indicate that when C/N was 20:1, the methane production rate was fastest, and Methanosaetaceae and Methanobacteriaceae were the primary methanogens within the Archaea. The richness and evenness of methanogenic bacteria was best with the highest T-RFLP diversity index of 1.627 in the six granular sludge samples. When C/N was reduced from 20:1 to 5:1, the methanogenic activity of SMD granules decreased gradually, and the relative quantities of methanogens decreased from 36.5% to 10.9%. The abundance of Methanosaetaceae in Archaea increased from 64.5% to 84.2%, while that of Methanobacteriaceae decreased from 18.6% to 11.8%, and the richness and evenness of methanogens decreased along with the T-RFLP diversity index to 1.155, suggesting that the community structure reflected the succession to an unstable condition represented by high nitrate concentrations.
Keywords Simultaneous methanogenesis and denitrification (SMD)      Methanogens      Community structure      Diversity index      Granular sludge     
Corresponding Author(s): Juanjuan Zhao,Jiane Zuo   
Issue Date: 03 August 2018
 Cite this article:   
Yujiao Sun,Juanjuan Zhao,Lili Chen, et al. Methanogenic community structure in simultaneous methanogenesis and denitrification granular sludge[J]. Front. Environ. Sci. Eng., 2018, 12(4): 10.
 URL:  
https://academic.hep.com.cn/fese/EN/10.1007/s11783-018-1067-2
https://academic.hep.com.cn/fese/EN/Y2018/V12/I4/10
Phase Time (d) C/N Loading rate
(kg/m3/d)		 Influent strength
(mg/L)		 Removal efficiency (%)
COD NO3−-N COD NO3−-N COD NO3−-N
I 0–46 – 10.0 – 3762.6 – 96.5 –
II 47–134 20:1 10.0 0.5 3762.6 188.5 97.7 100.0
III 135–152 15:1 10.1 0.7 3648.1 251.4 98.0 99.6
IV 153–219 10:1 10.7 1.1 3846.2 408.5 98.0 99.6
V 220–234 5:1 10.6 2.0 2216.9 406.4 96.0 100.0
Tab.1  Performances of UASB bioreactor under different C/N ratios
Fig.1  Changing of loading rate and removal efficiency of COD and NO3--N in the UASB bioreactor for 235 days
Fig.2  SEM images of sludge particles and microbial communities
Fig.3  Methane production rate in biomethane potential test of SMD granular sludge samples
Flora classification (Archaea) OTUs
quantity		 Clone quantity Content
(%)		 Representative strain
Methanosaetaceae 19 63 71.59 Methanosaeta concilli (X51423)
Methanobacteriaceae 9 20 22.73 Unculture Methanobacteriaceae
Archaeon (AB236056)
Methanospirillaceae 3 4 4.55 Unculture Methanoaspirillum sp.(AY692060)
Methanomicrobiaceae 1 1 1.14 Unculture Methanomicrobiaceae Archaeon (AB236985)
Tab.2  Clone results of Archaea in granules of sample D
Fig.4  Relative content of Archaea and Bacteria in SMD granular sludge samples
Fig.5  Relative content of Methanosaetaceae and Methanobacteriaceae in Archaea in SMD granular sludge samples
Reactions DrHm? (kJ/mol
e donor)		 DrSm?
(J/mol/K
e donor)		 DrGm? (kJ/mol
e donor)
Methanogenesis:
H2(g) + 0.25CO2(ao)→0.25CH4(g) + 0.5H2O(l) -29 -39 -17
CH3COOH(ao)→CH4(g) + CO2(ao) -1 31 -10
Denitrification:
H2(g) + 0.4H+(ao) + 0.4NO3−(ao)→0.2N2(g) + 1.2H2O(l) -130 -34 -164
CH3COO−(ao) + 2.6H+(ao) + 1.6NO3−(ao)→0.8N2(g) + 2CO2(ao) + 2.8H2O(l) -102 33 -155
Tab.3  Reactions occur in a SMD process
Fig.6  T-RFLP profiles of Archaea 16S rDNA PCR products amplified from SMD sludges at different C/N ratio
Sample No. T-RFLP diversity index
A 0.966
B 0.924
C 1.627
D 1.338
E 1.219
F 1.155
Tab.4  Diversity index T-RFLP of Archaea in six sludge samples
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