Protocol 07 National Development Centre Siam catfish Indonesia: Parent Maintenance, spawning and hatching Siam catfish (Pangasius)

Aquaculture Fisheries Development : The goal is to produce larvae that will one day be a prospective parent raised for breeding

A. Container cultivation requirement

1. Holding size pool maintenance 125-250 m2
2. Holding tank size of 2x5x0, 8 m3
3. Nets parent container 2x2x0, 8 m3 total of 4 pieces
4. Funnel hatching along the system
5. Fiber anesthetic volume size of 100 liters
6. mesh size 0.8 larvae shelter x0, 6x0, 5 m3
7. Fiber glass volume of 100 liters

 

B. Equipment

1. Volume of 3 ml syringe
2. plastic basin
3. plastic bucket
4. Fishing nets holding size of 1.5 x 2 x 1 m3
5. scoop parent
6. Weighing 60 kg parent; accuracy of 0.01 g
7. Scales eggs 2 kg; accuracy 0,001 g
8. Maximum-minimum thermometer
9. tagging equipment
10. microchip
11. microscope
12. Headcount minimum of 5 units
13. Pipette 5 ml volume
14. towel
15. Fish carrier bag measures 80 x 20 x 40 cm3
16. measuring cup
17. quill
18. Fiber anesthesia 100 liters
19. Water pumps for parent recovery
20. catheter
21. petri dish
22. 100 ml measuring cups

C. Material

1. parent feed 28-32% protein
2. Ovulation hormone (GnRH-a + Domperidone)
3. Materials anesthetic (clove oil: 25 ppm)
4. Sera solution (absolute ethanol, 40% formalin, and glacial acetic acid in the ratio 6:3:1)
5. Suspension of red soil (filtered first with subtle scoop
6. Sodium chloride (Na Cl 0.9%)
7. Tissue paper to dry the container and fish at the time of conception

D. Work Procedures
D.1. Maintenance of Fish parent

1. Parent kept in ponds at a density of 0.7 to 1 kg/m2 at a water depth of 1.5 to 2 m with male female ratio of 1: 2
2. The feed is 28-32% protein as much as 1-2% of the weight of the fish biomass per day
3. Feeding frequency 2-3 times a day.

D2.Selection of parent is ready spawned

1. Parent captured by netted.
2. Parent captured one by one and observed the state of the stomach and the urogenital.
3. Abdomen of female gonads mature enlarged, indicating there are eggs in it and urogenital red.
4. Parent mature female gonad put fiber anesthesia for egg samples taken by biopsy.
5. Egg samples were observed visually and microscopically with soaked in sera solution for 10 minutes.
6. Observations eggs are: Uniform egg size,Color yellowish white eggs, non-transparent (opaque), Core eggs are in a position in the middle or slightly to one side or stadia and stadia 2 (see apendix 1), Diameter eggs 1.0-1.2 mm.
7. Parent mature male gonad is characterized by abdominal massage anal direction.
8. If a thick white liquid had came out the male parent was transferred to parent incubation bath.
9. The selected female parent measured body length and body weight were weighed to determine the volume of hormones to be used, and then transferred to the incubation bath parent.
10. The selected female parent was transferred to parent incubation bath.
11. Parent females housed in the net in the incubation bath at a density of 1 fish per net.
12. Incubation for parent aerated Bath, water flowed continuously during the spawning
13. Male parent accommodated in a separate tub with a female parent

D3. Injecting hormones

1. Ovulation hormone is used GnRH-a + ​​Domperidone
2. Injection dose is 0.5 ml / kg female parent.
3. The injection is done 2 times at intervals of 6 hours.
4. Male parent was not injected with hormones.
5. The mother was anesthetized using anesthetic in fiber anesthesia
6. Anesthesia performed until the parent is not strong rebel (partially done)
7. Intramuscular injections made in the dorsal or pectoral fins intraperitonial below.
8. Checking ovulation time is 6 hours after the second injection.

D4. Fertilization

1. Checking ovulation time is 6 hours after the second injection
2. Anesthetized female parent (partially)
3. The abdomen of female genital pressed gently toward
4. When the eggs come out easily then the female parent is ready to stripping
5. Anesthetized female parent totally, gill cover to the tail and not moving
6. Parent females weighed his weight.
7. Water in the body of the female parent dried using a soft towel and tissue paper
8. Parent stripping by gently massaging the belly parent to the anal.
9. Eggs collected in plastics or stainless bowl.
10. Simultaneously, the male captured and drugged.
11. The eggs were processed for ovulation weighed
12. Stripping male parent who has fainted then sperm directly inserted in basin reservoirs eggs.
13. The addition of NaCl 0.9% in sperm-egg mixture and stir to coat the chicken until sperm covers the eggs evenly.
14. Fertilization is done by mixing water into the egg-sperm mixture and stir until evenly for 2-3 minutes.
15. Eggs are rinsed with clean water 3 times until the bubbles of sperm disappeared
16. The use of clay suspensions (see Appendix 2.) To remove the adhesive eggs
17. Stirring the mixture on top of using chicken feathers for 2-3 minutes until the eggs are separated from each other.
18. Eggs are rinsed again with clean water 3-4 times to rid the suspension of red soil.

 D5. Hatching in the hatching funnel

1. Preparing funnel hatching and installation.
2. Appropriated water quality requirements of water quality siamese catfish hatcheries (Table 1.)
3. Eggs inserted into hatch funnel a maximum of 500 ml per funnel.
4. Observation and calculation of the degree of fertilization performed 6 hours after fertilization (27-30 0C).
5. The eggs will hatch 16-26 hours after fertilization depending on the temperature of the hatching container
6. (27-30 0C).
7. The larvae hatch in the first 4-5 hours are used for the production of the prospective parent.
8. The larvae were harvested with a scoop shelter hapa smooth and collected in 100 liter fiber glass.
9. Larvae calculated using volumetric sampling methods *
10. Larvae ready to be distributed to the larval rearing container

* Volumetric sampling method

• Larvae collected in a container of known volume A (eg 100 liters) and strong aeration.
• Take a random sampling volume B (eg 100 ml)
• Calculate the larvae of volume B
• Repeat the second step several times
• Calculate the average number of larvae (C)
• The total number of larvae = A x ((1000 / B) xC) fish

Table 1. Water quality suitable for siamese catfish hatcheries media (SNI 01-6483.4-2000)
Temperature             : 27 C - 30 C
PH value                   : 6.5 to 8.5
Dissolved Oxygen   : > 5 mg / L

Apendix 1. The development of stadia egg or oocyte nucleus position at the end of oocyte maturation

Appendix 2. Preparation of clay suspensions

1. Prepare clay (red / orange / white) to taste (± 5 kg)
2. Clean dirt from the gravel, or grass
3. Soil dissolved in water
4. Filter the solution with a net scoop finely ground 2 layers
5. Filtered red soil deposited
6. Layer of water over the soil suspension removed to obtain a thick soil suspension
7. Keep the soil suspension in a cool (room temperature)
8. Suspension ground ready for use.

 Aquaculture Fisheries Development