Leffler Brunson
Development of a Multi-Trophic, Bioflocbased, Polyculture System for Production of Marine Shrimp, Red Drum, and Oysters John W. Leffler and Jeff F. Brunson
South Carolina Department of Natural Resources Marine Resources Research Institute Waddell Mariculture Center
Superintensive, Indoor, Minimal-exchange, Biofloc Shrimp Systems
235 m3 Advantages: Biosecurity High productivity Efficient & economical nitrogen control Supplemental food/nutrient recycling
Shrimp Feed
Shrimp Waste
Bacteria Cyanobacteria Green Algae
Chemoautotrophs
BIOFLOC
Photoautotrophs Heterotrophs
Diatoms Dinoflagellates Water Quality
Benefits Supplemental Nutrition
Solids NO3
Wastes
In an indoor, minimal-exchange, superintensive shrimp biofloc culture: Could the biofloc community of the shrimp culture expand to also process wastes generated by fish in the same system? Could shellfish utilize the biofloc as a food source? Could shellfish reduce the suspended solids to produce relatively clean water for fish production? Could shrimp, fish, and shellfish be grown at commercially viable rates?
Greenhouse-based, recirculating biofloc system for the simultaneous culture of Pacific white shrimp (Litopenaeus vannamei)
Red Drum (Sciaenops ocellatus)
Eastern oyster (Crassostrea virginica) Photos: Courtesy of SC Dept. of Natural Resources
Constructed four replicate polyculture systems each with total volume of 31 m3. Shrimp held in 30 m3 tanks filled to 22 m3. Provided main biofloc reservoir.
Fish were held in 7 m3 circular tanks with center drains.
Oysters held in 1 m3 rectangular troughs. Oysters in plastic mesh bags suspended from floats.
Included in each system was a 1 m3 settling tank to manage settleable solids.
Trials 1 and 2
Settling Tank
Oyster Tank
Flow rate ~ 20 L min-1 Shrimp Raceway
Red Drum Tank
Early Trials: Will shrimp system accommodate addition of fish? Shrimp
Red Drum
Oysters
4.4 ± 0.3 g
300 ± 92 g
7-10 cm
250 m-1
14.3 m-1
50 m-1
Length of trial
107 d
107 d
10 d
Survival
88%
90%
0%
1.0 g wk-1
7.4 g wk-1
0.0 g wk-1
Stocking size Stocking density
Growth rate
Final biomass supported = 134 kg; 4.3 kg m-3 Shrimp: Fish biomass ratio = 2:1
Shrimp and red drum found to be compatible in polyculture system.
Trial 2
NH3-N (mg/L)
Trial 1 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0
Ammonia-nitrogen
Shrimp Raceway
NO2-N (mg/L0
0.5
Fish Tank
Nitrite-nitrogen
When red drum were added, the Biofloc increased to process the additional nitrogen input.
0.4 0.3
NH3-N < 1.4 mg L-1
0.2 0.1
NO2-N ≤ 0.4 mg L-1
0.0
Fish Tank
Shrimp Raceway
Trial 3 Objectives
Trial 3
System modifications to improve movement of biofloc particles through all tanks Evaluate replicability of simultaneous systems in terms of water quality and production.
Diffuser manifolds for oxygen transfer and circular flow.
External standpipes removed daily to flush solids to shrimp tanks.
Trial 3
Trial 3 Initial Stocking System A
System B
System C
Shrimp mean weight ± SD (g)
11.6 ± 2.6
10.9 ± 3.2
12.6 ± 3.3
Fish mean weight ± SD (g)
559 ± 178
568 ± 163
562 ± 172
Oyster length ± SD (mm)
41 ± 7
38 ± 8
36 ± 9
Shrimp density (m-3)
275
275
275
Fish density (m-3)
14.3
14.3
14.3
30
30
30
Oyster density (m-3)
Shrimp fed Zeigler High Intensive 35 at 400 g d-1 per tank. Red drum were fed Zeigler Finfish Gold 42-16 slow sinking feed at 280 g d-1 per tank.
Trial 3
Replicability among Systems
A Shrimp A Fish B Shrimp B Fish C Shrimp
Replicate Systems - pH 7.8
1.4
21-Mar
14-Mar
7-Mar
28-Feb
21-Feb
14-Feb
7-Feb
Fish A Shrimp B
0.6
Fish B
0.4
21-Mar
14-Mar
7-Mar
Fish C
28-Feb
0.0
21-Feb
Shrimp C
14-Feb
0.2
7-Feb
21-Mar
14-Mar
7-Mar
28-Feb
21-Feb
14-Feb
6.8
Fish C
7-Feb
Shrimp C
31-Jan
7.0
24-Jan
Fish B
1.0 0.8
31-Jan
Shrimp B
7.2
Shrimp A
24-Jan
Fish A
7.4
1.2
17-Jan
Shrimp A
NO2-N (mg L-1)
1.6
17-Jan
pH
Replicate Systems - NO2-N
8.0
7.6
31-Jan
C Fish
24-Jan
1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0
17-Jan
NH3-N (mg L-1)
Replicate Systems - NH3-N
Replicability among Systems - Solids Replicate Systems - TSS
TSS (mg L-1)
1200.0 1000.0 800.0 600.0 400.0 200.0 0.0
Replicate Systems - Settleable solids 4.5
Shrimp A Fish A Shrimp B Fish B Shrimp C Fish C
4.0
Settleable solids (mL L-1)
1400.0
Trial 3
3.5
Settling tank input
3.0 2.5 2.0
Settling tank output
1.5 1.0 0.5 0.0 1/17/2013
2/17/2013
Replicability among Systems - Oysters
Trial 3
Oysters suffered 100% mortality within 10 days of stocking.
500
AO
400
BO
300
High solids load ?
CO
200
Replicate Systems TSS in Oyster Troughs
100 0
Inadequate food ?
TSS (mg L-1)
Total Chlorophyll (µg L-1)
Replicate Systems - Total Chlorophyll in Oyster Troughs
800 700 600 500 400 300 200 100 0
Oyster A Oyster B Oyster C
Trial 4 Objective: Permit Oyster Survival
Trial 4
Halted Trial 3 and Altered Shrimp & Fish Biomass to Create Different Biofloc Communities System
Shrimp Biomass (kg)
A
63.5
Fish System Biomass Total (kg) (kg)
48.8
B
88.9
63.2
C
33.4
26.6
D
Microbial Community
112.3 cyanobacterium Synechococcus few unicellular algae & diatoms, 152.1 filamentous bacteria, dinoflagelates, many rotifers unicellular algae, more diatoms, 60.0 dinoflagellates, many rotifers
Filtered seawater & unicellular green algae fertilized; no biomass
Trial 4 Oyster Survival? 25 oysters stocked in each trough for 10 days System
% Survival
Density
A
20
Medium
B
68
High
C
80
Low
D
88
No shrimp or fish
A - Dominated by cyanobacterium Synechococcus B - Filamentous bacteria, hetero. dinoflagellates C - No filamentous bacteria, hetero. dinoflagellates, more diatoms D - Unicellular green algae
Survival of Oysters in Serial Dilutions of Synechococcus sp. Bloom Water
No dilution
25% dilution
50% dilution
75% dilution
Trial 4
Trial 4
Impact of a Gymnodinium sp Bloom Change in Shrimp Weight during Gymnodinium Bloom in System C 27 25
grams
23 4/30/2013
21
5/22/2013
19 17 15 System A
System B
System C
Cysts so abundant that water was gray; Cysts settled into 1-2 mm layer in a 500 mL flask.
Impact of a Gymnodinium sp Bloom Change in Red Drum Percent Survival during Gymnodinium Bloom in System C 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0%
4/30/2013 5/22/2013
System A
System B
System C
Cysts very abundant on gills. Toxins probably killed the fish.
Preliminary Conclusions – BioFloc Polyculture System • Shrimp and red drum compatible. • Biofloc will process wastes of combined shrimp-fish biomass. • Oysters highly problematic. • Biomass loading influences biofloc community composition. • Outbreaks of cyanobacteria and heterotrophic dinoflagellates have significant negative impacts on growth and survival.
Can oysters survive and prosper? Can the biofloc community be made more stable? Could such a system approach profitability?
Please return for Biofloc Polyculture – Part 2
Trials 5 and 6!
presented by our next speaker:
Dr. Luis Poersch
Thank you! Acknowledgements
The Waddell Mariculture Center Team Deliah Arrington Matthew Brown Dr. Michael Denson Charles Hamilton Dr. Peter Kingsley-Smith Kevin Pitts Dr. Luis Poersch Jacob Richardson Robert Shumate Al Stokes
South Carolina Department of Natural Resources Marine Resources Research Institute Waddell Mariculture Center
Superintensive, Indoor, Minimal-exchange, Biofloc Shrimp Systems
235 m3 Advantages: Biosecurity High productivity Efficient & economical nitrogen control Supplemental food/nutrient recycling
Shrimp Feed
Shrimp Waste
Bacteria Cyanobacteria Green Algae
Chemoautotrophs
BIOFLOC
Photoautotrophs Heterotrophs
Diatoms Dinoflagellates Water Quality
Benefits Supplemental Nutrition
Solids NO3
Wastes
In an indoor, minimal-exchange, superintensive shrimp biofloc culture: Could the biofloc community of the shrimp culture expand to also process wastes generated by fish in the same system? Could shellfish utilize the biofloc as a food source? Could shellfish reduce the suspended solids to produce relatively clean water for fish production? Could shrimp, fish, and shellfish be grown at commercially viable rates?
Greenhouse-based, recirculating biofloc system for the simultaneous culture of Pacific white shrimp (Litopenaeus vannamei)
Red Drum (Sciaenops ocellatus)
Eastern oyster (Crassostrea virginica) Photos: Courtesy of SC Dept. of Natural Resources
Constructed four replicate polyculture systems each with total volume of 31 m3. Shrimp held in 30 m3 tanks filled to 22 m3. Provided main biofloc reservoir.
Fish were held in 7 m3 circular tanks with center drains.
Oysters held in 1 m3 rectangular troughs. Oysters in plastic mesh bags suspended from floats.
Included in each system was a 1 m3 settling tank to manage settleable solids.
Trials 1 and 2
Settling Tank
Oyster Tank
Flow rate ~ 20 L min-1 Shrimp Raceway
Red Drum Tank
Early Trials: Will shrimp system accommodate addition of fish? Shrimp
Red Drum
Oysters
4.4 ± 0.3 g
300 ± 92 g
7-10 cm
250 m-1
14.3 m-1
50 m-1
Length of trial
107 d
107 d
10 d
Survival
88%
90%
0%
1.0 g wk-1
7.4 g wk-1
0.0 g wk-1
Stocking size Stocking density
Growth rate
Final biomass supported = 134 kg; 4.3 kg m-3 Shrimp: Fish biomass ratio = 2:1
Shrimp and red drum found to be compatible in polyculture system.
Trial 2
NH3-N (mg/L)
Trial 1 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0
Ammonia-nitrogen
Shrimp Raceway
NO2-N (mg/L0
0.5
Fish Tank
Nitrite-nitrogen
When red drum were added, the Biofloc increased to process the additional nitrogen input.
0.4 0.3
NH3-N < 1.4 mg L-1
0.2 0.1
NO2-N ≤ 0.4 mg L-1
0.0
Fish Tank
Shrimp Raceway
Trial 3 Objectives
Trial 3
System modifications to improve movement of biofloc particles through all tanks Evaluate replicability of simultaneous systems in terms of water quality and production.
Diffuser manifolds for oxygen transfer and circular flow.
External standpipes removed daily to flush solids to shrimp tanks.
Trial 3
Trial 3 Initial Stocking System A
System B
System C
Shrimp mean weight ± SD (g)
11.6 ± 2.6
10.9 ± 3.2
12.6 ± 3.3
Fish mean weight ± SD (g)
559 ± 178
568 ± 163
562 ± 172
Oyster length ± SD (mm)
41 ± 7
38 ± 8
36 ± 9
Shrimp density (m-3)
275
275
275
Fish density (m-3)
14.3
14.3
14.3
30
30
30
Oyster density (m-3)
Shrimp fed Zeigler High Intensive 35 at 400 g d-1 per tank. Red drum were fed Zeigler Finfish Gold 42-16 slow sinking feed at 280 g d-1 per tank.
Trial 3
Replicability among Systems
A Shrimp A Fish B Shrimp B Fish C Shrimp
Replicate Systems - pH 7.8
1.4
21-Mar
14-Mar
7-Mar
28-Feb
21-Feb
14-Feb
7-Feb
Fish A Shrimp B
0.6
Fish B
0.4
21-Mar
14-Mar
7-Mar
Fish C
28-Feb
0.0
21-Feb
Shrimp C
14-Feb
0.2
7-Feb
21-Mar
14-Mar
7-Mar
28-Feb
21-Feb
14-Feb
6.8
Fish C
7-Feb
Shrimp C
31-Jan
7.0
24-Jan
Fish B
1.0 0.8
31-Jan
Shrimp B
7.2
Shrimp A
24-Jan
Fish A
7.4
1.2
17-Jan
Shrimp A
NO2-N (mg L-1)
1.6
17-Jan
pH
Replicate Systems - NO2-N
8.0
7.6
31-Jan
C Fish
24-Jan
1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0
17-Jan
NH3-N (mg L-1)
Replicate Systems - NH3-N
Replicability among Systems - Solids Replicate Systems - TSS
TSS (mg L-1)
1200.0 1000.0 800.0 600.0 400.0 200.0 0.0
Replicate Systems - Settleable solids 4.5
Shrimp A Fish A Shrimp B Fish B Shrimp C Fish C
4.0
Settleable solids (mL L-1)
1400.0
Trial 3
3.5
Settling tank input
3.0 2.5 2.0
Settling tank output
1.5 1.0 0.5 0.0 1/17/2013
2/17/2013
Replicability among Systems - Oysters
Trial 3
Oysters suffered 100% mortality within 10 days of stocking.
500
AO
400
BO
300
High solids load ?
CO
200
Replicate Systems TSS in Oyster Troughs
100 0
Inadequate food ?
TSS (mg L-1)
Total Chlorophyll (µg L-1)
Replicate Systems - Total Chlorophyll in Oyster Troughs
800 700 600 500 400 300 200 100 0
Oyster A Oyster B Oyster C
Trial 4 Objective: Permit Oyster Survival
Trial 4
Halted Trial 3 and Altered Shrimp & Fish Biomass to Create Different Biofloc Communities System
Shrimp Biomass (kg)
A
63.5
Fish System Biomass Total (kg) (kg)
48.8
B
88.9
63.2
C
33.4
26.6
D
Microbial Community
112.3 cyanobacterium Synechococcus few unicellular algae & diatoms, 152.1 filamentous bacteria, dinoflagelates, many rotifers unicellular algae, more diatoms, 60.0 dinoflagellates, many rotifers
Filtered seawater & unicellular green algae fertilized; no biomass
Trial 4 Oyster Survival? 25 oysters stocked in each trough for 10 days System
% Survival
Density
A
20
Medium
B
68
High
C
80
Low
D
88
No shrimp or fish
A - Dominated by cyanobacterium Synechococcus B - Filamentous bacteria, hetero. dinoflagellates C - No filamentous bacteria, hetero. dinoflagellates, more diatoms D - Unicellular green algae
Survival of Oysters in Serial Dilutions of Synechococcus sp. Bloom Water
No dilution
25% dilution
50% dilution
75% dilution
Trial 4
Trial 4
Impact of a Gymnodinium sp Bloom Change in Shrimp Weight during Gymnodinium Bloom in System C 27 25
grams
23 4/30/2013
21
5/22/2013
19 17 15 System A
System B
System C
Cysts so abundant that water was gray; Cysts settled into 1-2 mm layer in a 500 mL flask.
Impact of a Gymnodinium sp Bloom Change in Red Drum Percent Survival during Gymnodinium Bloom in System C 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0%
4/30/2013 5/22/2013
System A
System B
System C
Cysts very abundant on gills. Toxins probably killed the fish.
Preliminary Conclusions – BioFloc Polyculture System • Shrimp and red drum compatible. • Biofloc will process wastes of combined shrimp-fish biomass. • Oysters highly problematic. • Biomass loading influences biofloc community composition. • Outbreaks of cyanobacteria and heterotrophic dinoflagellates have significant negative impacts on growth and survival.
Can oysters survive and prosper? Can the biofloc community be made more stable? Could such a system approach profitability?
Please return for Biofloc Polyculture – Part 2
Trials 5 and 6!
presented by our next speaker:
Dr. Luis Poersch
Thank you! Acknowledgements
The Waddell Mariculture Center Team Deliah Arrington Matthew Brown Dr. Michael Denson Charles Hamilton Dr. Peter Kingsley-Smith Kevin Pitts Dr. Luis Poersch Jacob Richardson Robert Shumate Al Stokes
