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Plankton Culture for Feeding Larval Fish and Shellfish. LaDon Swann Mississippi-Alabama Sea Grant Consortium Auburn University. Shellfish Larval Culture. Larval Rearing. Trocophore Size 50-60 microns 6-12 hours post-fertilization D-Hinge or Straight Hinge Size 70-80 microns

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plankton culture for feeding larval fish and shellfish

Plankton Culture for Feeding Larval Fish and Shellfish

LaDon Swann

Mississippi-Alabama Sea Grant Consortium

Auburn University

larval rearing
Larval Rearing
  • Trocophore
    • Size 50-60 microns
    • 6-12 hours post-fertilization
  • D-Hinge or Straight Hinge
    • Size 70-80 microns
    • 24-48 hours post-fertilization
    • Bivalve Shell
    • Velum - Swimming and Feeding organ
larval rearing4
Larval Rearing
  • Veliger or Umbo Larvae
    • Size 80-300 microns
    • Stage last 8-12 days
    • Well defined umbo
  • Pediveliger or “Eyed” Larvae
    • Size >300 microns
    • Pigmented eye-spot
    • Muscular foot
phytoplankton production
Phytoplankton Production
  • Feeding Larvae
    • Cell Size 4-8 microns
    • Species
      • Isochrysis galbana
      • Chaetoceros gracilis
      • Nannochloris sp.
      • Chlorella sp.
      • Pavlova lutheri
pavlova lutheri
Pavlova lutheri
  • Morphology
    • Golden brown
    • Spherical with 2 flagella
    • 3-6 µm
  • Salinity
    • 8-32 ppt
  • Temperature
    • 11-26 °C
  • Culture media
    • Guillards f/2
  • Proximate Analysis
    • 52% Protein
    • 24% Carbs
    • 29% Fat
isochrysis galbana
Isochrysis galbana
  • Morphology
    • Tahiti (T-Iso strain)
    • Golden brown
    • Cells spherical with 2 flagella
    • 5-6 µm length, 2-4 µm wide
  • Salinity
    • 8-32 ppt
  • Temperature
    • 23 - 28°C
  • Culture media
    • Guillards f/2
  • Proximate Analysis
    • 47% Protein
    • 24% Carbs
    • 17% Fat
chaetoceros gracilis
Chaetoceros gracilis
  • Morphology
    • Golden brown diatom
    • Medium-size 12 µm wide, 10.5 µm long
    • Cells united in chains
  • Salinity
    • 26 - 32 ppt
  • Temperature
    • 28 - 30°C
  • Culture media
    • Guillards f/2 with Si
  • Proximate Analysis
    • 28% Protein
    • 23% Carbs
    • 9% Fat
plankton for shellfish
Plankton for Shellfish
  • Broodstock and Spat
    • Cell Size 10-24 microns
    • Species
      • Tetraselmis sp.
        • Green
      • Thalassiosra sp.
        • Diatom
tetraselmis sp
Tetraselmis sp.
  • Morphology
    • Ovoid green cells
    • 14 to 23 µm L X 8 µm W
    • 4 flagella
  • Salinity
    • 28-36 ppt
  • Temperature
    • 22-26°C
  • Culture media
    • Guillards f/2
  • Proximate Analysis
    • 55% Protein
    • 18% Carbs
    • 14% Fat
thalassiosra sp
Thalassiosra sp.
  • Morphology
    • Golden brown diatom
    • Cells united in chains
    • Barrel-shaped
    • Non-motile
    • 4 µm
  • Salinity
    • 26 – 32 ppt
  • Temperature
    • 22-29 °C
  • Culture media
    • Guillards f/2 with Si
  • Other characteristics
micro algae culture
Micro Algae Culture
  • Culture Water
  • Sterilization
  • Nutrient Enrichment
  • Inoculation
  • Cell Counts
  • Harvest and Feeding
  • Stock Culture
culture water
Culture Water
  • Sources
    • Seawater
    • Saltwater wells
    • Prepared seawater
  • Salinity
    • 26-32 ppt
sterilization
Sterilization
  • Methods
    • Heat Pasteurization
      • 80 C and cool naturally
    • Autoclave
    • Sodium Hypochlorite (bleach)
      • 0.5 ml/L (10 drops)
      • Neutralize: 10-15 ml sodium thiosulfate (248 g/L) per liter
    • Hydrochloric acid (muriatic)
      • 0.2 ml/L (4 drops)
      • Neutralize: Na2CO3 0.4-0.9 g/L
nutrient enrichment
Nutrient Enrichment
  • Guillard’s f/2
    • Part A and B
    • 0.5 ml/L each part
    • Na2Si03 for diatoms
inoculation
Inoculation
  • Culture vessels
    • 1,000 ml flask
    • 18.7 L (5 gal.) Carboy (glass)
    • 178 L (47 gal) Transparent Tank
  • Add enough algae to give a strong tint to the water
    • 100,000-200,000/ml
  • Lighting
    • Types
      • Sunlight
      • Fluorescent
      • VHO fluorescent
      • Metal halide
    • Highest Densities: 24/7
cell counts
Peak Algae Density

I. Galbana

10-12 million cells/ml

10-14 days

2 wk stability

T. pseudonana

4 million cells/ml

3 days

5 day stability

Hemacytometer

Count total in centermost 1 mm

Multiply by 10,000

Product = number/ml

Cell Counts

Motile cells should be killed

harvest and feeding
Algae Density

Wk 1 = 50,000 cells/ml

Wk 2+ = 100,000 cells/ml

Onset of spatting = 200,000/ml

Tank cleared in 24hrs

Larvae Density

5-10 larvae/ml

Harvest and Feeding

Liters to feed = (TD x V)/CD

TD = Target Density (1,000s/ml)

V = Volume of larval tank (thousands of L)

CD = Cell Density (millions/ml)

harvesting and feeding
Harvesting and Feeding
  • Batch
    • Total harvest occurs once or over several days
  • Semi-Continuous
    • Works well with diatoms
    • Part of the algae remains in the vessel
    • New media is added to replenish the algae removed
stock culture
Purchase pure strain

Avoid contamination

No aeration

Half filed container

Redundancy

Holding

Test tubes

Conical flasks

Transfer

1 drop/wk for T. pseudonana

1 drop/2 wk for I. galbana

Stock Culture
marine fish larval culture
Marine Fish Larval Culture
  • Relies on zooplankton
marine rotifer brachionus plicatilis
Marine Rotifer Brachionus plicatilis
  • Culture units
    • 40L plastic bags
    • 40L cone-bottomed tanks
  • Temperature 27-30 C
  • Salinity 26 ppt
rotifers
Rotifers
  • Laboratory production
    • 100 to 200+ mm size
    • 2-3 week life span
    • small size suitable as first food
feeding marine fish
Feeding Marine Fish
  • Rotifers
    • Typical first food in hatchery
    • Feed algae or yeast
    • Enrichment needed
artemia
Artemia
  • Feeding of older larvae
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