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Distinctive denitrifying capabilities leads to different N 2 O - - PowerPoint PPT Presentation

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Distinctive denitrifying capabilities leads to different N 2 O production in dPAO and dGAO cultures Anna Ribera-Guardia 1 , R. Marques 2 , C. Arangio 1 , M. Carvalheira 2 , A. Oehmen 2 , M. Pijuan 1 aribera@icra.cat 16th September 2016 N2O

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SLIDE 1

Anna Ribera-Guardia1, R. Marques2, C. Arangio1, M. Carvalheira2, A. Oehmen2, M. Pijuan1

aribera@icra.cat

Distinctive denitrifying capabilities leads to different N2O production in dPAO and dGAO cultures

16th September 2016

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SLIDE 2

N2O during denitrification

PO4- P

ATP

NO3

  • NO2
  • NO

N2 N2O

NiR Nor Nos NaR

ATP rNO3

  • Biomass

growth rNO2

  • rNO

rN2O

PHA

NO3

  • N2

ATP

PO4

3-

Poly- P PAO

?

PHA

NO3

  • ATP

Glycoge n

N2

GAO

? C-source

e- e- e- e-

PHA EBPR communities

Slower e- supply e- competition??

16th September 2016

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SLIDE 3
  • Explore the denitrification kinetics of two different dPAO and

dGAO cultures

  • Study the N2O accumulation potential of both cultures
  • Assess the occurrence of electron competition

Objectives

16th September 2016

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SLIDE 4

Materials and Methods: Bioreactors operation

  • 2 different SBR for dPAO and dGAO

cultures, respectively

  • Cycle: 6 hours
  • Anaerobic phase (1h 45 min)
  • Anoxic phase (2h)
  • Aerobic phase (1h 30 min)
  • Settling /decant phase/Idle (45 min)
  • Carbon source: Propionic and acetic

acids

  • SRT: 10 days
  • pH : 7.5  0.1

Phosphate media Waste activated sludge Argon Air Nitrogen media Carbon media O2 pH HCl 0.1 M

V = 2 L V = 6 L dGAO dPAO

16th September 2016

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SLIDE 5

Batch test type A B C D E F G Electron acceptors used NO3

  • NO2
  • N2O

NO3

  • NO2
  • NO3
  • NO3
  • N2O

N2O NO2

  • NO2
  • N2O

pH probe reservoir N2O microsensor Temperature probe

  • Sludge withdrawn from the end of the anaerobic phase of each reactor
  • 330 mL sealed reactor anoxic conditions
  • pH controlled at 7.5
  • Dissolved N2O online monitored by an N2O microsensor

Materials and Methods: Batch tests

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SLIDE 6

Results:SBRs operation

NO2- NO3- PO43- Acetate Propionate

3- Time (min) 100 200 300 Acetate, Propionate (mg COD/L); PO43- (mg P/L) 20 40 60 80 100 120 140 NO3-, NO2- (mg N/L) 5 10 15 20 25 30 35 Anaerobic Anoxic Aerobic Feed P and C Feed N

a

dPAO

16th September 2016

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SLIDE 7

Results:SBRs operation

NO2- NO3- PO43- Acetate Propionate

Time (min) 100 200 300 Acetate, Propionate (mg COD/L); PO43- (mg P/L) 20 40 60 80 100 120 140 NO3-, NO2- (mg N/L) 5 10 15 20 25 30 35 Anaerobic Anoxic Aerobic Feed P and C Feed N

b

Time (min) 100 200 300 Acetate, Propionate (mg COD/L); PO43- (mg P/L) 20 40 60 80 100 120 140 NO3-, NO2- (mg N/L) 5 10 15 20 25 30 35 Anaerobic Anoxic Aerobic Feed P and C Feed N

a

dPAO dGAO

16th September 2016

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SLIDE 8

Results:Microbial community

FISH PROBES Relative abundance dPAO-SBR PAO I 26.03 ± 4.75 % PAO II 15.42 ± 2.82 % PAOMIX 42.40 ± 8.32 % GAOMIX 22.93 ± 4.41 % DFImix, DFIImix and DFIII 4.17 ± 0.16 % dGAO-SBR GAOMIX 55.60 ± 1.86 % DFImix, DFIII and DFIV 6.33 ± 0.24 % DFIImix 13.20 ± 0.88 % PAOMIX 14.30 ± 1.52 %

16th September 2016

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SLIDE 9

Results:single electron acceptor

  • N2O accumulates for both cultures

dPAOs Time (min)

10 20 30 40

NO3- , NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

0.0 0.1 0.2 0.3 0.4 0.5

A

dGAOs Time(min)

10 20 30 40

NO3-, NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

0.0 0.1 0.2 0.3 0.4 0.5

A

NO2- NO3- N2O

  • Nitrite accumulates for dGAOs

16th September 2016

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SLIDE 10

Results:single electron acceptor

  • dPAOs prefer nitrite over nitrate
  • dGAOs prefer to consume nitrate

dPAOs Time (min)

10 20 30 40

NO3- , NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

0.0 0.1 0.2 0.3 0.4 0.5

A

dGAOs Time(min)

10 20 30 40

NO3-, NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

0.0 0.1 0.2 0.3 0.4 0.5

A

NO2- NO3- N2O

  • NO2- inhibits N2O reduction in dGAOs

Time (min)

10 20 30 40

NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

1 2 3 4 5 6

B

dPAOs Time(min) 10 20 30 40 NO2- (mg N/g VSS) 5 10 15 20 25 30 35 N2O (mg N/g VSS) 1 2 3 4 5 6

B

dGAOs

16th September 2016

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SLIDE 11

Results:single electron acceptor

  • dPAOs consume N2O faster than dGAOs

dPAOs Time (min)

10 20 30 40

NO3- , NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

0.0 0.1 0.2 0.3 0.4 0.5

A

dGAOs Time(min)

10 20 30 40

NO3-, NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

0.0 0.1 0.2 0.3 0.4 0.5

A

NO2- NO3- N2O

Time (min)

10 20 30 40

NO2- (mg N/g VSS)

5 10 15 20 25 30 35

N2O (mg N/g VSS)

1 2 3 4 5 6

B

dPAOs Time(min) 10 20 30 40 NO2- (mg N/g VSS) 5 10 15 20 25 30 35 N2O (mg N/g VSS) 1 2 3 4 5 6

B

dGAOs Time (min)

10 20 30 40 N20 (mg N/g VSS) 5 10 15 20 25 30 35

C dPAOs Time (min)

10 20 30 40

N2O (mg N/g VSS)

5 10 15 20 25 30 35

C dGAOs

16th September 2016

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SLIDE 12

Results: multiple e- acceptors

dPAOs Tests A B C D E F G NOx red rates (mg N/gVSS·h) 5 10 15 20 25 30 35

NO3- NO2- N2O

dGAOs Tests A B C D E F G NOx red rates (mg N/gVSS·h) 5 10 15 20 25 30 35

NO3- NO2- N2O 16th September 2016

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SLIDE 13
  • dPAOs have higher denitrifying capacity
  • Nitrate reduction rates in dPAOs are constant in all tests
  • dGAOs presented higher N2O accumulated/N-reduced

Results: multiple e- acceptors

dPAOs Tests A B C D E F G NOx red rates (mg N/gVSS·h) 5 10 15 20 25 30 35

NO3- NO2- N2O

dGAOs Tests A B C D E F G NOx red rates (mg N/gVSS·h) 5 10 15 20 25 30 35

NO3- NO2- N2O 16th September 2016

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SLIDE 14
  • No electron competition

Results: electron competition

dPAOs Tests

A B C D E F G

Electron consumption rate (mmole/gVSSh

1 2 3 4 5 6 7 8 9

Nar Nir Nor Nos

dGAO Tests

A B C D E F G

Electron consumption rate (mmole/gVSSh

1 2 3 4 5 6 7 8 9

Nar Nir Nor Nos

  • Higher e- consumption rates in dPAOS higher reduction rates

16th September 2016

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SLIDE 15

 N2O accumulation was higher in dGAOs than in dPAOs and

intensified when nitrite was present  No electron competition was detected  dPAOs have higher denitrifying capacity than dGAOs

Conclusions

16th September 2016

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SLIDE 16

Acknowledgements

Project CTM 2011-27163 and CTM 2015-66892-R Ramon y Cajal RYC-2009-04959 PhD grant BES-2012-052753 Acciones IntegradasPRI-AIBPT-2011-1232) Luso-Espanhola action E-61/12 COST action ES1202 (Water 2020) PTDC/AAC-AMB/12058/2010, UID/Multi/04378/2013 , PhD grant SFRH/BD/74515/2010

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SLIDE 17

16th September 2016

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SLIDE 18

Distinctive denitrifying capabilities leads to different N2O production in dPAO and dGAO cultures

Anna Ribera-Guardia1, R. Marques2, C. Arangio1, M. Carvalheira2, A. Oehmen2, M. Pijuan1

aribera@icra.cat

Thank you very much for your attention

16th September 2016