Appendix 4:  Schedule of animals' daily diet and food preparation

Animals' daily diet

Food is supplied to the growing animals according to their age (size) and filtration efficiency by varying the amount of food components together with the seawater flow rate (turnovers). The following table describes the schedule that we apply during the first month of life. The feeding increments are approximatively 30% at the 20^th day and 50% at the 30^th day (after this time there are no substantial changes in the filtration efficiency).

Age (days)	Seawater turnovers per day	food: Spirulina	food: Progression	food: Revolution
5	10	1.5 g	0.5 g	2.0 ml
15	7	1.5 g	0.5 g	2.0 ml
20	7	2.0 g	0.7 g	2.5 ml
30	7	3.0 g	1.0 g	4.0 ml

Food used in our study was selected by performing several trials in which various foods were tested in different combinations.

Food preparation

1. Prepare three 250 ml plastic bottles (mix-bottles) with 150 ml of clean seawater.
2. Weigh out the amount of the three different foods and add each into a mix-bottle with seawater (Fig. 15).
3. Shake vigorously until the mixtures have a uniform appearance.
4. The Spirulina algae solution should be allowed to hydrate for 5 min before use.
5. Prepare two 5 liter bottles (food bottle) half-filled with clean seawater and place them on the stirrers.
6. Add the Spirulina food solution to one food bottle, and the Progression and Revolution solutions to the other food bottle.
7. If desired, pass the mixtures through an appropriately sized micron net prior to adding to the bottles.
8. Fill the food bottles up to 5 liters.
9. Set the peristaltic pump tubes between the bottles and the "food tank," and switch on the pumps.

NOTES:

1. The use of two food bottles is for practical reasons, in order to obtain a more dilute food solution; but it is also possible to use a single 10-liter bottle in which to prepare the food mixtures.

2. The bore of a peristaltic tube is selected so as to obtain a flow of food solutions into the water ("food tank") of either 5 or 10 liters per 24 h, depending on whether 2 or 1 food bottles are used.

3. The feeds (provided by Salt Creek, Inc.) used are :

   - Jade Spirulina algae

   - Progression 1

   - Revolution 1