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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 l.jpg

Plankton Culture for Feeding Larval Fish and Shellfish

LaDon Swann

Mississippi-Alabama Sea Grant Consortium

Auburn University



Larval rearing l.jpg
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 l.jpg
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


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Phytoplankton Production

  • Feeding Larvae

    • Cell Size 4-8 microns

    • Species

      • Isochrysis galbana

      • Chaetoceros gracilis

      • Nannochloris sp.

      • Chlorella sp.

      • Pavlova lutheri


Pavlova lutheri l.jpg
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 l.jpg
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 l.jpg
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 l.jpg
Plankton for Shellfish

  • Broodstock and Spat

    • Cell Size 10-24 microns

    • Species

      • Tetraselmis sp.

        • Green

      • Thalassiosra sp.

        • Diatom


Tetraselmis sp l.jpg
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 l.jpg
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 l.jpg
Micro Algae Culture

  • Culture Water

  • Sterilization

  • Nutrient Enrichment

  • Inoculation

  • Cell Counts

  • Harvest and Feeding

  • Stock Culture


Culture water l.jpg
Culture Water

  • Sources

    • Seawater

    • Saltwater wells

    • Prepared seawater

  • Salinity

    • 26-32 ppt


Sterilization l.jpg
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 l.jpg
Nutrient Enrichment

  • Guillard’s f/2

    • Part A and B

    • 0.5 ml/L each part

    • Na2Si03 for diatoms


Inoculation l.jpg
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 l.jpg

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 l.jpg

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 l.jpg
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 l.jpg

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 l.jpg
Marine Fish Larval Culture

  • Relies on zooplankton


Marine rotifer brachionus plicatilis l.jpg
Marine Rotifer Brachionus plicatilis

  • Culture units

    • 40L plastic bags

    • 40L cone-bottomed tanks

  • Temperature 27-30 C

  • Salinity 26 ppt


Rotifers l.jpg
Rotifers

  • Laboratory production

    • 100 to 200+ mm size

    • 2-3 week life span

    • small size suitable as first food


Feeding marine fish l.jpg
Feeding Marine Fish

  • Rotifers

    • Typical first food in hatchery

    • Feed algae or yeast

    • Enrichment needed


Artemia l.jpg
Artemia

  • Feeding of older larvae



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