anammox for nutrient nitrogen removal from sludge liquors fiel… · mineralization. o 2. e-o. 2....
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Anammox for nutrient nitrogen removal from sludge liquors
Dept. Chemical & Environmental EngineeringUniversity of Arizona
Jim Field, Guangbin Li and Reyes Sierra
Industrial Partners: Pima County Regional Wastewater Reclamation Department (PCRWRD)
Los Angeles County Sanitation District (LACSD)
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The N Cycle
2
NH4+N2
NO2-
NO3-
Org-Nmineralizationas
sim
ilatio
n
O2
O2e-
e-
N-fixation
nitrification-denitrification
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Costs N Removal
Conventional Nutrient N Removal Costs
Nitrification:
Denitrification: NO3- + 5 e- eq → 0.5 N2
64 g O2 / mol N
40 g COD/ mol N
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Conventional Nutrient N Removal
DenitrificationAOBNitrification
NOB
N2
NH4+ NO3-
AOB
NOB
O2 COD
ammonia nitrate
dinitrogen gas
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The Missing Link in the N Cycle
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NH4+N2
NO2-
NO3-
Org-N
anammox
mineralizationas
sim
ilatio
n
O2
O2e-
e-
N-fixation
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Anammox
Discovered in Late 1980’s in The Netherlands
Simplified Stoichiometry
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N2NH4+ NO2-ammonia nitrate dinitrogen gas
+
Anaerobic Ammonium Oxidation
Nitrate was mixed with anaerobic effluent at Gist Brocadus to oxidize hydrogen sulfide and anammox was discovered due to NH4
+ in wastewater
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Costs and Savings of AnammoxAnammox Reaction (Detailed Stoichiometry)
NH4+
(100% N)
57% flow
43% flow
57% less O2
no O2 needed for anammox
NitritationAnammox
N2 89% NNO3
- 11% NAOB
NO2-
no COD needed for anammox
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Anammox is combined with AOB as a technology
O2 No COD
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Issues with Anammox
pH optima are critical
Sensitive to Substrate (NO2-) Toxicity
H2S is very toxic
Sensitivity depends heavily on physological status cells
Low pH exacerbates NO2- stress
High pH slowly deactivates anammox
Slow growthe rate
Arizona has high sulfate levels in municipal wastewater 7
Doubling times range from 5 to 15 d
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Toxicity of NO2-
starved/NO2- pre-exposure
NO2- pre-exposure
active
starved/active
Physiological status
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Is it Possible to Cultivate Anammox from Return Activated Sludge (RAS)?
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RAS anammox enrichment
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Enrichment Procedure
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minerals + Medium:
NH4+ + NO2
-
NH4+ + NO2
- NH4+ + NO2
- 10% v/v--------- spikes --------- --------------- transfers ------------
90% vol new medium
10% v/v
time
Establish Enrich
Controls: Just NH4+ (no NO2
-) “no nitrite control”Just NO2
- (no NH4+) “endogenous control”
NO2- + NH4
+ (no inoculum) “abiotic control”
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Enrichment ExperimentExample: Anammox Enrichment (RAS, Ina Rd)
NH4+ Graph
abiotic
no nitrite control
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Enrichment ExperimentExample: Anammox Enrichment (RAS, Ina Rd)
NO2- Graph
abiotic
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Activity Experiment RAS Anammox Enrich.
0.0
1.0
2.0
3.0
4.0
5.0
0 5 10 15 20 25 30 35Time (hr)
NO
2- con
cent
ratio
n (m
M)
0.0
1.0
2.0
3.0
4.0
0 5 10 15 20 25 30 35Time (hr)
NH
4+ con
cent
ratio
n (m
M)
0.0
1.0
2.0
3.0
4.0
0 5 10 15 20 25 30 35Time (hr)
N2 c
once
ntra
tion
(mm
ole
L liq
uid-1
)
0.0
0.2
0.4
0.6
0 5 10 15 20 25 30 35Time (hr)
NO
3- con
cent
ratio
n (m
M)
NO2-
NH4+
N2
NO3-
abiotic
endogenous
abiotic
no nitrite control
no nitrite control
no nitrite control
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N-Balance RAS Anammox Enrichment
Final Balance
NO2- = 3.89 mM
NH4+ = 2.75 mM
Sum N = 6.64 mM
Consumed
NO3- = 0.57 mM
N2 = 3.02 mmol gas/Lliq
Sum N = 6.61 mM
Produced
Ratios
Measured: NO2-/NH4
+ = 1.40
Theoretical: NO2-/NH4
+ = 1.3214
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DNA Evidence of RAS Anammox EnrichmentPCR forward primer Pla46F, reverse primer AMX-820R
Lane 1: MW markers
Lane 2: demiwater
Lane 3: anammox enrichment, Univ. Santiago de Compostela, Spain
Lane 4: anammox sludge Bioreactor Paques, The Netherlands
Lane 7: Activated Sludge High O2
Lane 8: RAS Ina Rd, original sample
Lane 5: RAS oxidation ditch
Lane 6: RAS Ina Rd, anammox enrichment
In theory PCR product 774 bp
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Conclusions Enrichment RAS
First consistent evidence of Anammox activity = 45 d
Starting Material = Ina Road, RAS
Anammox the dominant reaction = 65 d
Genus of enriched anammox = Brocadia
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Doubling time = 9.2 d
Details of enrichment: Sun et al. Chemosphere 2011, 84:1262-1269
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Expanded Granular Sludge Bed (EGSB)
17influent
effluent
N2 gas
sludge bed
gas cap
settler
baffles
sludge granulegas bubble
weir3 phaseseparator
recirculationeffluent
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Expanded Granular Sludge Bed (EGSB)
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Expanded Granular Sludge Bed (EGSB)
HRT = 5.11 hInfl NO2- = 29 mM (1340 mg NO2-/L)Infl NH4+ = 25 mM (450 mg NH4+ /L)pH range effluent = 7.2-7.7
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Statistics Field-Sierra Research Group with the Anammox Process
7 oral presentations (inter)national conferences
3 research projects
3 PhD dissertations, 3 MS theses, 4 post docs
15 journal articles on Anammox (incl. 1 pending)
NSF-CBET $337,340 (2013-2017)USDA (US-Egypt STDF) $180,000 (2010-2012)WSP $83,450 (2008-2009)
WEF/IWA Nutrient Removal and Recovery, Jul. 12th, 2016 Denver
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Questions
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Biochemistry
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Cell Morphology
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Bioaugmentation Strategy
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Strategy to Recover Bioreactor Experienceing NO2
- Stress
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Bioaugmentation Strategy
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Experience Research Group Field-Sierra with the Anammox Process
Molecular ecology
Reactor technology experience
Operating anammox under “optimal conditions”
Experience with upward-flow anaerobic sludge bed for annamoxMembrane bioreactor
Preventing inhibitionReversing reactor failure
Clone librariesPrimer sets for functional genes
Granular biomass
pH optimumStress markers
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What Field-Sierra Group Can do for Pima Co.
Operate bench-scale reactors with Tres Rios reject water
Develop, test and monitor strategies for rapid cultivation of anammox biomass with reject water
Advise and provide monitoring program pilot scale reactors
Compare reactor technology strategies
Monitoring program
Advise full-scale plant
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pH Optima versus Biomass Types
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Toxicity of H2S
Granular biomass
Suspended biomass
IC50 suspended = 0.03 mM (1 ppm)
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pH Optima versus Biomass Types
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Toxicity of H2S
Granular biomass
Suspended biomass
IC50 suspended = 0.03 mM (1 ppm)
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