Step 1 — Prepare Incubation Solution
- Prepare a saline solution using sea salt at a concentration of 28–30 ppt
- Alternatively, use filtered seawater (preferably UV or ozone treated)
Adjust pH to 7.9–8.5
- If pH is lower, add 2 g Sodium Bicarbonate (NaHCO₃) per liter
- Maintain temperature at 27–29°C
Step 2 — Prepare Decapsulation Solution
For 454 g of cysts:
- Use 1.5 L Sodium Hypochlorite (NaOCl)
- Add 34–50 ml NaOH (saturated solution)
Standard ratio:
1 L bleach (10% chlorine)
- 23–33 ml NaOH
Step 3 — Hydration
- Place cysts in freshwater
- Apply strong aeration for 1 hour
- Hydration is complete when cysts change from biconcave to spherical shape
Step 4 — First Decapsulation
- Transfer hydrated cysts into 0.75 L decapsulation solution
- Stir continuously for 25–40 minutes
- Monitor temperature closely, as the reaction is exothermic
- If temperature rises, cool the tank externally with cold water
Step 5 — Separation and Washing
- Separate cysts using a 100-micron mesh filter
- Rinse thoroughly with clean seawater
Step 6 — Second Decapsulation
- Repeat decapsulation for 40–50 minutes
- Continue until cysts change color from white to orange
Step 7 — Activation
- Add Sodium Erythorbate (C₆H₇O₆Na) to the incubation solution
Dosage:
- 0.45 g per 1 g cysts
- Approximately 200 g per 454 g cysts
- Add 30 minutes before incubation
Step 8 — Incubation
- Add cysts at a density of 2 g per liter
- Start strong aeration
- Provide 1500–2000 lux illumination
- Maintain incubation for 24 hours
Step 9 — Harvest
- Stop aeration
- Remove shells from the surface
- Collect nauplii using filtration
- Washing is optional
Nauplii are ready for use