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Vs. Aquaculture Live or Prepared Feeds Vs. Aquaculture Live or Prepared Feeds

Man-made. . . • Pellets or flakes. • Dry pellets the norm (uniform nutrition. Man-made. . . • Pellets or flakes. • Dry pellets the norm (uniform nutrition. ) • Disadvantages: rapid sinking, unless extruded. • Semi-moist pellets: soft, high quality. • Disadvantage: expensive!! difficult to store in bulk.

What do we use? ? • Manufactured fish feeds are made from a variety What do we use? ? • Manufactured fish feeds are made from a variety of ingredients: fish meal being the main one! • Alternatives are always welcome. • Many times it is easier to buy off the shelf. • What if you’re fish are unique, or won’t eat man made diets, or they just can’t? !

What is common here? • Small size • Incomplete development • Lack of digestive What is common here? • Small size • Incomplete development • Lack of digestive ability! Images of a tautog as it develops

Feeding Larval Fish: Plankton Culture Feeding Larval Fish: Plankton Culture

Phytoplankton Production • Feeding Larval Fish – Cell Size 4 -8 microns – Species Phytoplankton Production • Feeding Larval Fish – Cell Size 4 -8 microns – Species • Isochrysis galbana • Chaetoceros gracilis • Nannochloris sp. • Chlorella sp. • Pavlova lutheri

Pavlova lutheri • Morphology – Golden brown – Spherical with 2 flagella – 3 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 • Morphology – – Tahiti (T-Iso strain) Golden brown Cells spherical with 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 • Morphology – Golden brown diatom – Medium-size 12 µm wide, 10. 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 • Broodstock and Spat – Cell Size 10 -24 microns – Plankton for Shellfish • Broodstock and Spat – Cell Size 10 -24 microns – Species • Tetraselmis sp. – Green • Thalassiosra sp. – Diatom

Tetraselmis sp. • Morphology – Ovoid green cells – 14 to 23 µm L 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. • Morphology – – – Golden brown diatom Cells united in chains 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 • • Culture Water Sterilization Nutrient Enrichment Inoculation Cell Counts Harvest Micro Algae Culture • • Culture Water Sterilization Nutrient Enrichment Inoculation Cell Counts Harvest and Feeding Stock Culture

Culture Water • Sources – Seawater – Saltwater wells – Prepared seawater • Salinity Culture Water • Sources – Seawater – Saltwater wells – Prepared seawater • Salinity – 26 -32 ppt

Sterilization • Methods – Heat Pasteurization • 80 C and cool naturally – Autoclave 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: Na 2 CO 3 0. 4 -0. 9 g/L

Nutrient Enrichment Nutrients Na. NO 3 Conc. • Guillard’s f/2 (mg/l Seawater) – Part Nutrient Enrichment Nutrients Na. NO 3 Conc. • Guillard’s f/2 (mg/l Seawater) – Part A and B 75 Na. H 2 PO 4. H 2 O 5 Na 2 Si. O 3. 9 H 2 O 30 Na 2 C 10 H 14 O 8 N 2. H 2 O (Na 2 EDTA) 4. 36 Co. Cl 2. 6 H 2 O 0. 01 Cu. SO 4. 5 H 2 O 0. 01 Fe. Cl 3. 6 H 2 O 3. 15 Mn. Cl 2. 4 H 2 O 0. 18 Na 2 Mo. O 4. 2 H 2 O 0. 006 Zn. SO 4. 7 H 2 O 0. 022 Thiamin HCl 0. 1 Biotin 0. 0005 B 12 0. 0005 – 0. 5 ml/L each part – Na 2 Si 03 for diatoms

Inoculation • Culture vessels – 1, 000 ml flask – 18. 7 L (5 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 Cell Counts • Peak Algae Density – I. Galbana • 10 -12 million cells/ml • 10 -14 days • 2 wk stability – T. pseudonana • Hemacytometer – Count total in centermost 1 mm – Multiply by 10, 000 – Product = number/ml • 4 million cells/ml • 3 days • 5 day stability Motile cells should be killed

Harvest and Feeding • Algae Density • Larvae Density – 5 -10 larvae/ml – Harvest and Feeding • Algae Density • Larvae Density – 5 -10 larvae/ml – Wk 1 = 50, 000 cells/ml – Wk 2+ = 100, 000 cells/ml – Onset of spatting = 200, 000/ml • Tank cleared in 24 hrs Liters to feed = (TD x V)/CD TD = Target Density (1, 000 s/ml) V = Volume of larval tank (thousands of L) CD = Cell Density (millions/ml)

Harvesting and Feeding • Batch – Total harvest occurs once or over several days 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

Marine Fish Larval Culture • Relies on zooplankton Marine Fish Larval Culture • Relies on zooplankton

Marine Rotifer Brachionus plicatilis • Culture units – 40 L plastic bags – 40 Marine Rotifer Brachionus plicatilis • Culture units – 40 L plastic bags – 40 L cone-bottomed tanks • Temperature 27 -30 C • Salinity 26 ppt

Rotifers • Laboratory production – 100 to 200+ mm size – 2 -3 week Rotifers • Laboratory production – 100 to 200+ mm size – 2 -3 week life span – small size suitable as first food

Feeding Marine Fish • Rotifers – Typical first food in hatchery – Feed algae Feeding Marine Fish • Rotifers – Typical first food in hatchery – Feed algae or yeast – Enrichment needed

Artemia • Feeding of older larvae Artemia • Feeding of older larvae

Artemia Preparation • Brine shrimp eggs/cysts are used globally as a food for small Artemia Preparation • Brine shrimp eggs/cysts are used globally as a food for small fish. • Eggs/cysts: dry=dormant for years!! • Cysts can be used unhatched, but it’s risky. • Can kill small fish.

Decapsulation • Sometimes decapsulation is needed (remove shell): chlorine (household bleach), leaving the unhatched Decapsulation • Sometimes decapsulation is needed (remove shell): chlorine (household bleach), leaving the unhatched baby brine shrimp protected in a membrane. • Besides making the harvest of the hatched brine shrimp easier, this process also: -sterilizes the eggs -higher percentage of hatching -feed unhatched eggs to fish -decapsulated eggs can be hatched later (stored in the refrigorator)

Equipment • A 3 -gallon container with clear sides • 1 pound of brine Equipment • A 3 -gallon container with clear sides • 1 pound of brine shrimp eggs • 1 gallon of non-fragranced household bleach (5% chlorine) • Brine shrimp net or filter • Saturated brine solution*

Procedure • Soak 1 pound of eggs in 1 gallon of fresh water for Procedure • Soak 1 pound of eggs in 1 gallon of fresh water for 1 hour gently aerating the eggs. • After soaking, add 1 gallon of non-fragranced liquid household beach (5% chlorine) and reduce aeration. • Wait till eggs turn orange. • Strain contents through a brine shrimp net (or filter), and rinse in fresh water. • Store in saline solution for up to a month.

Use of copepods. . . Use of copepods. . .

 • Larval rearing systems are often labor intensive. • Skilled workers are needed • Larval rearing systems are often labor intensive. • Skilled workers are needed to maintain larval food culture. • Sometimes special equipment or systems are needed for small fish.