james m. ebeling, ph.d. aquaculture engineer
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An Engineering Analysis of the Stoichiometry An Engineering Analysis of the Stoichiometry of Autotrophic, Heterotrophic Bacterial Control of Autotrophic, Heterotrophic Bacterial Control
of Ammonia-Nitrogen in Zero-Exchange Productionof Ammonia-Nitrogen in Zero-Exchange Production
James M. Ebeling, Ph.D. James M. Ebeling, Ph.D. Aquaculture EngineerAquaculture Engineer
Michael B. Timmons, Ph.D.Michael B. Timmons, Ph.D.ProfessorProfessor
Dept. of Bio. & Environ. Eng.Dept. of Bio. & Environ. Eng.Cornell UniversityCornell University
James J. Bisogni, Ph.D.James J. Bisogni, Ph.D.ProfessorProfessor
School of Civil & Environ. Eng.School of Civil & Environ. Eng.Cornell UniversityCornell University
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
IntroductionIntroduction
Ammonia-nitrogen
NH4+
-N NO2--N
Ammonia Oxidizing Bacteria
Photoautotrophic(green-water systems)
HeterotrophicHeterotrophic(zero-exchange systems)
NH4+
-N C5H7O2N
BacteriaBacteria
Autotrophic(fixed-cell bioreactors)
NO2- -N NO3
--NNitrite Oxidizing Bacteria
NH4+
-N CC106106HH263263OO110110NN1616PP
AlgaeAlgae
Cinorganic / N Ratio Corganic / N Ratio
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
““New Paradigm”New Paradigm”
Zero-exchange Systems “Belize System”Zero-exchange Systems “Belize System”
ShrimpShrimp – – high health, selectively bred SPF stockhigh health, selectively bred SPF stock
FeedFeed – – high protein feeds combined with carbon supplementationhigh protein feeds combined with carbon supplementation
Water managementWater management – – zero water exchange, recycling water between cropszero water exchange, recycling water between crops
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
““New Paradigm”New Paradigm”
Zero-exchange Systems “Belize System”Zero-exchange Systems “Belize System”
ShrimpShrimp – – high health, selectively bred SPF stockhigh health, selectively bred SPF stock
FeedFeed – – high protein feeds combined with carbon supplementationhigh protein feeds combined with carbon supplementation
Water managementWater management – – zero water exchange, recycling water between cropszero water exchange, recycling water between crops
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??? Understanding of the ‘Removal System’??? Understanding of the ‘Removal System’
• PhotoautotrophicPhotoautotrophic• AutotrophicAutotrophic• HeterotrophicHeterotrophic• Some Combination!Some Combination!
Impact on Water Quality!!!!Impact on Water Quality!!!! Management Strategies!Management Strategies!
““New Paradigm” New Paradigm” ???? ????
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Ammonia ProductionAmmonia Production
In general:In general:
PPTANTAN = F * PC * 0.092 = F * PC * 0.092
where:where: PPTANTAN = Production rate of total ammonia nitrogen, (kg/day) = Production rate of total ammonia nitrogen, (kg/day)
F = Feed rate (kg/day)F = Feed rate (kg/day)
PC = protein concentration in feed (decimal value)PC = protein concentration in feed (decimal value)
For marine shrimp:
PTAN = F * PC * 0.144
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Nitrogen Removal Pathways
Extensive Pond
• Photoautotrophic
• Autotrophic
• Heterotrophic
• Other Mysterious Ways
NH4+-N CO2 Alkalinity
VSSAlgae
VSSAuto
VSSHetero
CO2
Alkalinity
O2
Corganic
NO3--N
TraceNutrients
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Nitrogen Removal Pathways
Intensive Pond • Photoautotrophic• Autotrophic
• Heterotrophic
• Other Mysterious Ways
NH4+-N CO2 Alkalinity
VSSAlgae
VSSAuto
VSSHetero
CO2
Alkalinity
O2
Corganic
NO3--N
TraceNutrients
Algae Based Systems
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Nitrogen Removal Pathways
Recirculation Systems
• Photoautotrophic
• Autotrophic• Heterotrophic
• Denitrification
NH4+-N O2 Alkalinity
VSSAlgae
VSSAuto
VSSHetero
Alkalinity
CO2
Corganic
NO3--N
TraceNutrients
NO2--N
Fixed-film Bioreactors
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Nitrogen Removal Pathways
Zero-exchange
• Photoautotrophic
• Autotrophic
• Heterotrophic
• Denitrification
NH4+-N O2 Alkalinity
VSSAlgae
VSSAuto
VSSHetero
Alkalinity
CO2
Corganic
NO3--N
TraceNutrients
Suspended Growth Systems
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PhotoautotrophicPhotoautotrophic (algal based systems)(algal based systems)
Biosynthesis of saltwater Biosynthesis of saltwater algaealgae:: Nitrate as nitrogen sourceNitrate as nitrogen source
16 16 NONO33-- + 124 + 124 COCO22 + 140 H + 140 H22O + O + HPOHPO44
2-2- → →
CC106106HH263263OO110110NN1616PP + 138 + 138 OO22 + 18 + 18 HCOHCO33--
Ammonia as nitrogen sourceAmmonia as nitrogen source
16 16 NHNH44++ + 92 + 92 COCO22 + 92 H + 92 H22O + 14 O + 14 HCOHCO33
-- + + HPOHPO442-2- → →
CC106106HH263263OO110110NN1616PP + 106 + 106 OO22
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PhotoautotrophicPhotoautotrophic (algal based systems)(algal based systems)
Consumes C organic C inorganic N
Consumables Stoichiometry (g) (g) (g) (g)
NH4+-N 1.0 ----- ----- 1.0
Carbon Dioxide 18.07 g CO2/ g N 18.07 ----- 4.93 -----
Alkalinity 3.13 g Alk/ g N 3.13 ----- 0.75 -----
Yields C organic C inorganic N
Products Stoichiometry (g)(g) (g) (g)
VSSAlgae 15.85 g VSSA / g N 15.85 5.67 ----- 1.0
Oxygen 15.14 g O2/ g N 15.14 ----- ----- -----
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Autotrophic - NitrificationAutotrophic - Nitrification
Biosynthesis of Biosynthesis of Autotrophic bacteriaAutotrophic bacteria:: NH4
+ + 1.83 O2 + 1.97 HCO3- →
0.024 C5H7O2N + 0.976 NO3- + 2.9 H2O + 1.86 CO2
The major factors affecting the rate of nitrification include:
• ammonia-nitrogen and nitrite-nitrogen concentration • carbon/nitrogen ratio• dissolved oxygen • pH• temperature• alkalinity• salinity
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Autotrophic - NitrificationAutotrophic - Nitrification
Consumes C organic C inorganic N
Consumables Stoichiometry (g) (g) (g) (g)
NH4+-N 1.0 ----- ----- 1.0
Alkalinity 7.05 g Alk/ g N 7.05 ----- 1.69 -----
Oxygen 4.18 g O2/ g N 4.18 ----- ----- -----
Yields C organic C inorganic N
Products Stoichiometry (g) (g) (g) (g)
VSSA 0.20 g VSSA / g N 0.20 0.106 ----- 0.025
NO3--N 0.976 g NO3
--N /g N 0.976 ----- ----- 0.976
CO2 5.85 g CO2/ g N 5.85 ----- 1.59 -----
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Heterotrophic BacteriaHeterotrophic Bacteria
The major factors affecting the rate of nitrification include:
• ammonia-nitrogen• carbon/nitrogen ratio• dissolved oxygen • pH• temperature• alkalinity• salinity
Biosynthesis of Biosynthesis of Heterotrophic bacteriaHeterotrophic bacteria:: NH4
+ + 1.18 C6H12O6 + HCO3- + 2.06 O2 →
C5H7O2N + 6.06 H2O + 3.07 CO2
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Heterotrophic BacteriaHeterotrophic Bacteria
Consumes C organic C inorganic N
Consumables Stoichiometry (g) (g) (g) (g)
NH4+-N 1.0 ----- ----- 1.0
C6H12O6 15.17 g Carbs/ g N 15.17 6.07 ----- -----
Alkalinity 3.57 g Alk/ g N 3.57 ----- 0.86 -----
Oxygen 4.71 g O2/ g N 4.71 ----- ----- -----
Yield C organic C inorganic N
Products Stoichiometry (g) (g) (g) (g)
VSSH 8.07 g VSSH / g N 8.07 4.29 ----- 1.0
CO2 9.65 g CO2/ g N 9.65 ----- 2.63 -----
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Impact of C/N Ratio
Autotrophic Heterotrophic
inorganic carbon as alkalinity
organic carbon from the feed
(109 g C organic /kg feed)@ 35% protein
organic carbon from the feed plus supplemental carbohydrates
C/N Ratio
Clabile /N ~ 2.2
C/N ~ 8-10
35.6% Heterotrophic 64.4 % Autotrophic
Clabile /N ~ 6.2
C/N ~ 12-16
Clabile /N ~ 0
Corganic/N ~ small
100% Heterotrophic 100 % Autotrophic
(Recirculation System withexcellent solids removal)
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StoichiometryStoichiometry
Photoautotrophic System
16 NO3- + 124 CO2 + 140 H2O + HPO4
2- → C106H263O110N16P + 138 O2 + 18 HCO3-
50.4 g N * 15.8 g VSS/ g N
= 800 g VSSphotoautotrophic
0.063 gN/gVSSA 0.358 gC/gVSSA
50.4 g NVSS 286 g CVSS
Conversion of 1 kg of feed @ 35% protein
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating AquacultureConversion of 1 kg of feed @ 35% protein
Consumes C organic C inorganic N
Consumables Stoichiometry (g) (g) (g) (g)
NH4+-N 50.4 ----- ----- 50.4
Carbon Dioxide 18.07 g CO2/ g N 911 ----- 249 -----
Alkalinity 3.13 g Alk/ g N 158 ----- 37.9 -----
Yields C organic C inorganic N
Products Stoichiometry (g) (g) (g) (g)
VSSAlgae 15.85 g VSSA / g N 800 286 ----- 50.4
O2 15.14 g O2/ g N 763 ----- ----- -----
Photoautotrophic (Pond intensive system)
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StoichiometryStoichiometry
50.4 g N * 0.20 g VSS/ g N
= 10.1 g VSSautotrophic
0.124 gN/gVSSA 0.531 gC/gVSSA
1.25 g NVSS 5.35 g CVSS
+ 49.2 g NO3-N + 80.1 g CO2
Autotrophic System
Conversion of 1 kg of feed @ 35% protein
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Autotrophic (Intensive Recirculation System)
Consumes C organic C inorganic N
Consumables Stoichiometry (g) (g) (g) (g)
NH4+-N 50.4 ----- ----- 50.4
Alkalinity 7.05 g Alk/ g N 355 ----- 85.6 -----
Oxygen 4.18 g O2/ g N 211 ----- ----- -----
Yields C organic C inorganic N
Products Stoichiometry (g) (g) (g) (g)
VSSA 0.20 g VSSA / g N 10.1 5.35 ----- 1.25
NO3--N 0.976 g NO3
--N /g N 0.976 ----- ----- 49.2
CO2 5.85 g CO2/ g N 295 ----- 80.1 -----
85.6 g CAlk / 50.5 g N C/N ratio of 1.7 and TOC is very, very small
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Zero-exchange System (no Carbon Supplementation)
Heterotrophic and Autotrophic Components
1 kgfeed * 0.36 kg BOD/kg feed * 0.40 kg VSS/ kg BOD =
= 144 g VSSheterotrophic
0.124 gN/gVSSH 0.531 gC/gVSSH
17.9 g NVSS 76.5 g CVSS
+ 47.1 g CCO2 = 123.6 g C
108.2 g Cfeed 15.4 g CAlkalinityAssume that the heterotrophic bacteria out compete the autotrophic bacteria.
Heterotrophic Component: Organic Carbon from Feed
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Zero-exchange System (no Carbon Supplementation)
Heterotrophic and Autotrophic Components
Excess Ammonia-nitrogen:50.4 g NH3-N - 17.9 g NVSS = 32.5
g NA
Autotrophic Component: Inorganic Carbon Alkalinity
32.5 g N * 0.20 g VSS/ g N
= 6.5 g VSSautotrophic
0.124 gN/gVSSA 0.531 gC/gVSSA
0.80 g NVSS 3.45 g CVSS
+ 31.7 g NO3-N + 55.4 g CAlk
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Heterotrophic Bacteria Consumes C organic C inorganic N
Stoichiometry (g) (g) (g) (g)
NH4+-N 0.356 * NT 17.9 ----- ----- 17.9
C6H12O6 feed 15.17 g Carbs/ g N 272 108.9 ----- -----
Alkalinity 3.57 g Alk/ g N 63.9 ----- 15.4 -----
Autotrophic Bacteria Consumes C organic C inorganic N
Stoichiometry (g) (g) (g) (g)
NH4+-N 0.644 * NT 32.5 ----- ----- 32.5
Alkalinity 7.05 g Alk/ g N 229.1 ----- 55.4 -----
C organic C inorganic N
Total Consumed Consumes (g) (g) (g)
NH4+-N 50.4 g N ----- ----- 50.4
C6H12O6 272 g Carbs 108.9 ----- -----
Alkalinity 293 g Alk ----- 70.8 -----
Zero-exchange System (no Carbon Supplementation)
Heterotrophic and Autotrophic Components
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Heterotrophic Bacteria Yields C organic C inorganic N
Stoichiometry (g) (g) (g) (g)
VSSH 8.07 g VSSH / g N 144 76.5 ----- 17.9
CO2 9.65 g CO2/ g N 174 ----- 47.4 -----
Autotrophic Bacteria Yields C organic C inorganic N
Stoichiometry (g) (g) (g) (g)
VSSA 0.20 g VSSA / g N 6.5 3.45 ----- 0.81
NO3--N 0.976 g NO3-N/g N 31.7 ----- ----- 31.7
CO2 5.85 g CO2/ g N 189 ----- 51.7 -----
C organic C inorganic N
Total Products Yields (g) (g) (g)
VSS 150.5 g VSS 80.0 ----- 18.7
NO3--N 31.7 g NO3-N ----- ----- 31.7
CO2 363.4 g CO2 ----- 99.1 -----
Zero-exchange System (no Carbon Supplementation)
Heterotrophic and Autotrophic Components
460 g Cfeed / 50.5 g N C/N ratio of 9
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Zero-exchange System (no Carbon Supplementation)
Heterotrophic and Autotrophic Components
83%
62%
50%
64%
69%72%
75%77%
23%25%
28%31%
36%
41%
17%
38%
50%
59%
0%
20%
40%
60%
80%
100%
12.4% 15% 20% 25% 30% 35% 40% 45% 50% 55%
Heterotrophic Bacteria
Autotrophic Bacteria
Percent removal of ammonia-nitrogen by Heterotrophic or Autotrophic Process as a function of % Protein
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Excess Ammonia-nitrogen:50.4 g NH3-N - 17.9 g NVSS = 32.5 g NA
32.5 g N * 8.07 g VSS / g N
= 262 g VSSheterotrophic
0.124 gN/gVSSH 0.531 gC/gVSSH
32.5 g NVSS 139 g CVSS
+ 85 g C CO2 = 225 g C
197 g Cs 28 g CAlkalinity
Carbon Supplement
Zero-exchange System (Carbon Supplementation)
Carbohydrate is 40% Carbon 492 g carbs
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture(460 g Cfeed + 197 g Ccarb ) / 50.5 g N C/N ratio of 13
Consumes C organic C inorganic N
Consumables Stoichiometry (g) (g) (g) (g)
NH4+-N 50.4 ----- ----- 50.4
C6H12O6 15.17 g Carbs/ g N 765 306 ----- -----
Alkalinity 3.57 g Alk/ g N 180 ----- 43.3 -----
Oxygen 4.71 g O2/ g N 237 ----- ----- -----
Yield C organic C inorganic N
Products Stoichiometry (g) (g) (g) (g)
VSSH 8.07 g VSSH / g N 407 216 ----- 50.4
CO2 9.65 g CO2/ g N 487 ----- 133 -----
Zero-exchange System (Carbon Supplementation)
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Supplemental Carbohydrate as percentage of feed ratefor heterotrophic metabolism of ammonia-nitrogen to microbial biomass
6%
17%
28%
38%
60%
71%
82%
93%
49%
0%
20%
40%
60%
80%
100%
15 20 25 30 35 40 45 50 55
Feed Protein (%)
Supp
lem
enta
l Car
bohy
drat
e .
(% o
f fee
d) .
Zero-exchange System (Carbon Supplementation)
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Research Trial #1 – C/N RatioResearch Trial #1 – C/N Ratio
StockingStocking 3.6 gm mean weight3.6 gm mean weight 150 shrimp / m150 shrimp / m22
Treatment (Sucrose)Treatment (Sucrose) ControlControl 50 % of Feed Rate50 % of Feed Rate 100% of Feed Rate100% of Feed Rate
4x12 Juvenile Production Tanks
DosageDosage 100% carbon Demand100% carbon Demand 200% of carbon Demand200% of carbon Demand
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Research SystemResearch Systemy =0.90 gms/wk
R2 = 0.98
2.0
4.0
6.0
8.0
10.0
12.0
0 10 20 30 40 50 60 70Days
Mea
n W
eigh
t (gm
s) .
Control
50% C Demand
100% C Demand
4x12 System with sludge settling tank,automatic feeders, bacterial floc
Weekly Growth – about 0.9 gm/wk
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Water Quality ParametersWater Quality Parameters
• Dissolved OxygenDissolved Oxygen• SalinitySalinity• TemperatureTemperature• pHpH• AlkalinityAlkalinity• TSS/TVSTSS/TVS
• TANTAN• NONO22 –N –N
• NONO33 –N –N
• Total NitrogenTotal Nitrogen• Total Organic CarbonTotal Organic Carbon
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Water Quality SummaryWater Quality Summary
DO Temp Salinity pH TAN NO2-N NO3-N Alkalinity
(mg/L) (Cº) (ppt) (mg/L) (mg/L) (mg/L) (mg/L)
Control 6.1 29.5 4.8 7.78 1.15 0.13 54.7 183StDev: 0.4 0.5 0.4 0.20 1.06 0.15 29.0 49
50% of Feed 5.7 29.8 4.5 8.15 1.06 0.38 7.7 328StDev: 0.9 0.9 0.4 0.14 0.26 1.0 3.3 22
100% of Feed: 5.3 29.4 4.7 8.19 1.36 0.60 1.9 360StDev: 1.5 0.2 0.2 0.18 0.81 1.1 0.8 24
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Water Quality Parameters - Water Quality Parameters - Dissolved OxygenDissolved Oxygen
4x12 Tanks: Dissolved Oxygen (mg/L)
0.0
2.0
4.0
6.0
8.0
10.0
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Dis
solv
ed O
xyge
n (m
g/L
) .
Control
50% Feed
100% Feed
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Water Quality Parameters - Water Quality Parameters - TemperatureTemperature
4x12 Tanks: Water Temperature (Deg. C)
27.0
28.0
29.0
30.0
31.0
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Tem
pera
ture
(Deg
C)
.
Control
50% Feed
100% Feed
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Water Quality Parameters - Water Quality Parameters - pHpH
4x12 Tanks: pH
7.00
7.50
8.00
8.50
9.00
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pH
Control
50% Feed
100% Feed
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Water Quality Parameters - Water Quality Parameters - TANTAN
4x12 Tanks: Ammonia-nitrogen (mg/L)
0.00
0.50
1.00
1.50
2.00
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TA
N (m
g/L
)
Control
50% Feed
100% Feed
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Water Quality Parameters – Water Quality Parameters – NONO22- N- N
4x12 Tanks: Nitritie-nitrogen (mg/L)
0.00
0.10
0.20
0.30
0.40
0.50
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Nit
rite
-nit
roge
n (m
g/L
) .
Control
50% Feed
100% Feed
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Water Quality Parameters – Water Quality Parameters – NONO33- N- N
4x12 Tanks: Nitrate-nitrogen (mg/L)
0
20
40
60
80
100
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Nit
rate
-nit
roge
n (m
g/L
) . Control
50%Feed
100% Feed
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Water Quality Parameters – Water Quality Parameters – AlkalinityAlkalinity
4x12 Tanks: Alkalinity (mg/L CaCO3)
50
100
150
200
250
300
350
400
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Alk
alin
ity
(mg/
L C
aCO
3) .
Control
50% Feed
100% Feed
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Solids ManagementSolids Management
Solids Management:Solids Management:
• Settling conesSettling cones
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Settling BasinsSettling Basins
Sedimentation: AdvantagesSedimentation: Advantages• Simplest technologiesSimplest technologies• Little energy inputLittle energy input• Relatively inexpensive to install and operateRelatively inexpensive to install and operate• No specialized operational skillsNo specialized operational skills• Easily incorporated into new or existing facilitiesEasily incorporated into new or existing facilities
18
D)(gV
2pp
s
Sedimentation: DisadvantagesSedimentation: Disadvantages• Low hydraulic loading ratesLow hydraulic loading rates• Poor removal of small suspended solidsPoor removal of small suspended solids• Large floor space requirementsLarge floor space requirements• Resuspension of solids and leechingResuspension of solids and leeching
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Settling BasinsSettling Basins
Design to minimize turbulence:Design to minimize turbulence:
vs = 0.0015 ft/sec
Q = flow (gpm)
vo = 0.00076 ft/sec
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Autotrophic/Heterotrophic ModelAutotrophic/Heterotrophic Model
ControlTank "A"
Initial Final TSS Removed % RemovedDay (mg/L) (mg/L) (gms)24 466 210 666 55%43 487 260 590 47%56 462 263 517 43%
Mean: 591
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50% of FeedTank "C"
Initial Final TSS Removed % RemovedDay (mg/L) (mg/L) (gms)11 379 218 419 42%22 441 134 798 70%34 449 206 632 54%42 441 242 517 45%52 482 218 686 55%60 485 313 447 35%
Mean: 583
Heterotrophic Model Heterotrophic Model (50% feed)(50% feed)
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100% of FeedTank "B"
Initial Final TSS Removed % RemovedDay (mg/L) (mg/L) (gms)
7 381 165 562 57%13 448 220 591 51%20 604 192 1071 68%25 569 304 689 47%28 490 219 705 55%32 445 208 616 53%35 422 187 611 56%40 507 185 837 64%45 520 264 666 49%49 490 254 614 48%54 463 350 294 24%58 673 587 224 13%62 579 338 627 42%
Mean: 623
Heterotrophic Model Heterotrophic Model (100% feed)(100% feed)
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TSS Production ModelTSS Production Model
Solids Production Model:Solids Production Model: autotrophic/heterotrophicautotrophic/heterotrophic
heterotrophicheterotrophic
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Autotrophic/Heterotrophic ModelAutotrophic/Heterotrophic Model
• allocated the daily feed organic carbon to heterotrophic bacterial production, allocated the daily feed organic carbon to heterotrophic bacterial production,
• calculated VSScalculated VSSHH, [, [VSSVSS
HH = feed g/m = feed g/m33 day * 0.36 g BOD/g feed * 0.40 g VSS day * 0.36 g BOD/g feed * 0.40 g VSSHH / g BOD] / g BOD]
• calculated amount of ammonia-nitrogen assimilated in the VSScalculated amount of ammonia-nitrogen assimilated in the VSSHH, [, [TANTAN
HH = 0.123 * VSS = 0.123 * VSSHH] ]
• subtracted TANsubtracted TANHH from the daily TAN from the daily TAN
feedfeed produced, produced,
[TAN[TANfeedfeed = feed g/m = feed g/m33 day * (0.35 * 0.16 * 0.9)] day * (0.35 * 0.16 * 0.9)]
• allocated excess ammonia-nitrogen to autotrophic bacterial consumption, allocated excess ammonia-nitrogen to autotrophic bacterial consumption,
[TAN[TANAA= TAN= TAN
feedfeed – TAN – TANHH]]
• determined VSSdetermined VSSA A [VSS[VSS
AA = TAN = TANAA * 0.20 g VSS * 0.20 g VSS
AA/g N]/g N]
• calculated Total VSS and TSS.calculated Total VSS and TSS.
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
Heterotrophic Model Heterotrophic Model (50% feed)(50% feed)
• allocated the daily feed carbon to heterotrophic bacterial production, allocated the daily feed carbon to heterotrophic bacterial production,
• calculated VSScalculated VSSHH, [, [VSSVSSHH = feed g/m = feed g/m33 day * 0.36 g BOD/g feed * 0.40 g VSS day * 0.36 g BOD/g feed * 0.40 g VSSHH / g BOD] / g BOD]
• calculated amount of ammonia-nitrogen sequestered in the VSScalculated amount of ammonia-nitrogen sequestered in the VSSHH, [, [TANTANHH = 0.123 * VSS = 0.123 * VSSHH] ]
• subtracted from the daily TANsubtracted from the daily TANfeedfeed produced, [ produced, [TANTANfeedfeed = feed g/m = feed g/m33 day * (0.35 * 0.16 * 0.9)] day * (0.35 * 0.16 * 0.9)]
• allocated excess ammonia-nitrogen to additional heterotrophic bacterial production, allocated excess ammonia-nitrogen to additional heterotrophic bacterial production,
[[TANTANH+H+= TAN= TANfeedfeed – TAN – TANH H ]]
• determined VSSdetermined VSSH+H+ [VSS[VSSH+H+ = 8.07 g VSS = 8.07 g VSSHH/g N * g N]/g N * g N]
• calculated Total VSS and TSS.calculated Total VSS and TSS.
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
Heterotrophic Model Heterotrophic Model (100% feed)(100% feed)
•allocated the daily feed carbon to heterotrophic bacterial production,
•calculated VSSH, [VSSH = feed g/m3 day * 0.36 g BOD/g feed * 0.40 g VSSH / g BOD]
•assumed all of the sucrose carbon was converted into bacterial biomass(sufficient nitrogen available)
•determined VSSdetermined VSSH+ H+ [VSSH+ = g sucrose/m3 day * 0.56 g VSSH/g sucrose]
•calculated Total VSS and TSS.
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
4x12 System: Total Suspended Solids Management - Control
y = 14.7 mg/L day
R2 = 0.92
y = 14.3 mg/L day
R2 = 0.93
y = 14.4 mg/L day
R2 = 0.87
y = 13.8 mg/L day
R2 = 0.94
50
150
250
350
450
550
0 7 14 21 28 35 42 49 56 63
Days
TS
S (
mg/
L)
Autotrophic/Heterotrophic ModelAutotrophic/Heterotrophic Model
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
4x12 System Total Suspended Solids - 50% of Feed
y = 29.1mg/L dayR2 = 0.93
y = 23.5 mg/L day R2 = 0.88
y = 27.4 mg/L dayR2 = 0.93
y = 35.5 mg/L dayR2 = 0.99
y = 21.8 mg/L dayR2 = 0.91
y = 38.1 mg/L dayR2 = 0.95
50
150
250
350
450
550
0 7 14 21 28 35 42 49 56 63
Days
TS
S (
mg/
L)
Heterotrophic Model Heterotrophic Model (50% feed)(50% feed)
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
4x12 System Total Suspended Solids - 100% of Feed
y = 53.7 mg/L day9
R2 = 0.96
y = 87.8 mg/L day
R2 = 0.96
y = 77.1 mg/L day
R2 = 0.99
y = 81 mg/L day
R2 = 0.99
y = 52.2 mg/L day
R2 = 0.98
50
200
350
500
650
0 7 14 21 28 35 42 49 56 63
Days
TS
S (
mg/
L)
Heterotrophic Model Heterotrophic Model (100% feed)(100% feed)
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Water Quality Management – C/N RatioWater Quality Management – C/N Ratio
0.0
2.0
4.0
6.0
8.0
10.0
7/18/04 7/25/04 8/1/04 8/8/04
Suga
r (k
g)
0.0
0.5
1.0
1.5
Nit
rite
(mg/
L-N
) .
Sugar (kg)
Nitrite (mg/L)
Impact of carbon supplementation (or lack of) on nitrite-nitrogen
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
Water Quality Parameters - Water Quality Parameters - TOCTOC
TOC for 4x12 System: Control, 50% and 100% Feed
y = 1.10 mg/L day
R2 = 0.97
Control
0
20
40
60
80
100
120
5/17/04 5/27/04 6/6/04 6/16/04 6/26/04 7/6/04 7/16/04
TO
C (m
g/L
-C)
.
Control
50% Feed
100% Feed
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
Water Quality Parameters - Water Quality Parameters - TNTN
0
50
100
150
200
250
300
0 10 20 30 40 50 60 70
Nit
roge
n (m
g/L
-N)
.
T otal Nitrogen - Model
T otal Nitrogen - FeedT otal Nitrogen - Experimental Data
Measured T otal Nitrogen
0
50
100
150
200
250
0 10 20 30 40 50 60 70
Nit
roge
n (m
g/L
-N)
.
Total Nitrogen - Model
Total Nitrogen - FeedTotal Nitrogen - Experimental Data
Measured Total Nitrogen
Mass balance on nitrogen for the autotrophic/heterotrophic system without carbon supplementation and with periodic harvesting of excess bacterial biomass
Impact of carbon supplementation at 50% of the feed as sucrose on the system with excess bacterial biomass and nitrogen being periodically removed from the system
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
Conclusions
Further work is needed to characterize the impact on Further work is needed to characterize the impact on production system performance at various C/N ratios.production system performance at various C/N ratios.
Alternative forms of Carbon need to be evaluated for Alternative forms of Carbon need to be evaluated for effectiveness and economics.effectiveness and economics.
Fundamental research is needed on carbon assimilation and Fundamental research is needed on carbon assimilation and conversion efficiency for heterotrophic bacteria.conversion efficiency for heterotrophic bacteria.
Development of optimal strains of bacteria for zero-exchange Development of optimal strains of bacteria for zero-exchange systems. systems.
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
Conclusions
““New Paradigm”New Paradigm” Zero-exchange systemsZero-exchange systems Mixed-cell racewaysMixed-cell raceways SPF, low salinity growoutSPF, low salinity growout
““Engineering Sustainability”Engineering Sustainability”
Organic CarbonOrganic Carbon + Nitrogen + Nitrogen → → Bacterial BiomassBacterial Biomass
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
AcknowledgementsAcknowledgements
Research was supported by the Agriculture Research ServiceResearch was supported by the Agriculture Research Service of the United States Department of Agriculture, of the United States Department of Agriculture,
under Agreement No. 59-1930-1-130under Agreement No. 59-1930-1-130 and Magnolia Shrimp LLC, Atlanta Georgiaand Magnolia Shrimp LLC, Atlanta Georgia
with special thanks to Miami Aqua-culture, Inc., Dan Spotts.with special thanks to Miami Aqua-culture, Inc., Dan Spotts.
Opinions, conclusions, and recommendations are of the authorsOpinions, conclusions, and recommendations are of the authors and do not necessarily reflect the view of the USDA.and do not necessarily reflect the view of the USDA.
All experimental protocols involving live animals were in complianceAll experimental protocols involving live animals were in compliance with Animal Welfare Act (9CFR) and have been with Animal Welfare Act (9CFR) and have been
approved by the Freshwater Institute Animal Care and Use Committee.approved by the Freshwater Institute Animal Care and Use Committee.
6th International Conference on Recirculating Aquaculture6th International Conference on Recirculating Aquaculture
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