< Rotifer Culture Protocol 1>

1. Material
  - live rotifer for inoculation
  - cold chlorella(CH) solution for rotifer food
  - 1L plastic bottle for culture bottle - Fig. A
  - heater for water bath at 28 ℃
  - aeration (6mm tubing, air stone, air pump)
  - culture solution: KW10(10ppt sea salt/RO)
  - Cage with a mesh(#350) – Fig. B

2. Condition
  - water bath 28℃
  - aeration




3. Time table (refer to Fig.A)
  ① 1st day (09:00)
     <1 STEP> 1L plastic bottle for culture bottle.
       A ◀ KW10 600mL + C 200mL ▶ add CH 2mL
       B ◀ KW10 600mL + C 200mL ▶ add CH 2mL
       C ◀ KW10 600mL + C 200mL ▶ add CH 2mL
     <2 STEP> put the 1 airstone with 6mm tubing into the 1L plastic bottle.
     <3 STEP> aeration
  ② 2~3 day (09:00) (once a day)
       A ▶ add CH 2mL
       B ▶ add CH 2mL
       C ▶ add CH 2mL
  ③ 4 day (09:00)
       A, B ▶ recovery using #350 (Fig.B) and feeding.
       C ▶ inoculate to A', B', C' (repeat ①)
            A' ◀ C 200mL + KW10 600mL ▶ add CH 2mL
            B' ◀ C 200mL + KW10 600mL ▶ add CH 2mL
            C' ◀ C 200mL + KW10 600mL ▶ add CH 2mL
  ④ Repeat ②~③

Warning.
1. cold fresh water chlorella is better than cold sea water chlorella.
2. may use mesh cage(#350) 2~3 times.
3. must check feeding time depending on chlorella concentration, color from green to light brown.
(Refer to Fig.C) : over-dose CH may inhibit proliferation of rofiter.
4. rotifer and fresh water chlorella supplier : http://www.aquasm.com/    4Liter/pk.
5. rotifer must be used in a day.
6. must be cleaned by a neutral detergent.
7. fry must be raised in 1ppt sea salt/RO.




※ Rofiter culture Tips.
  1. increase recovery efficiency and shorten the culture period using high density inoculation.
      (at least more than 1,500ea.)
  2. check solution color change from strong green to transparent brown.
  3. check the density of rotifer every morning using spoid or microscope.