Hybridization method of Sinocyclocheilus grahami and carp

11547,097

17/204302

March 17, 2021

A hybridization method of Sinocyclocheilus grahami and carp, which comprises parent fish cultivation, artificial insemination, fish egg incubating and fry feeding, strictly controls environmental parameters and feed types of parent fish cultivation, simulates good living conditions for S. grahami and carp, is beneficial for wild S. grahami and carp to adapt to the artificial breeding environment quickly, realizes natural spermatogenesis and oviposition in artificial environment without using the oxytocin, and provides sperm and eggs both in quality and quantity for artificial hybridization. At the same time, the key parameters during incubation (including incubating water level, water temperature, water flow velocity, pH, dissolved oxygen and sterilization of fertilized eggs, etc.) are also defined to provide suitable environmental conditions for the successful incubation of fertilized eggs.

Kunming Institute of Zoology, Chinese Academy of Sciences (Yunnan Province, CN)

KUNMING INSTITUTE OF ZOOLOGY, CHINESE ACADEMY OF SCIENCES (Yunnan Province, CN)

1. A hybridization method of S. grahami and carp, wherein comprising the following steps: placing the S. grahami and carp in culture ponds respectively for parent fish culture, wherein environmental parameters in culture ponds of S. grahami parent fish are as follows: water temperature is in a range of 15-22° C., water flow velocity is in a range of 5-20 cm/s, the concentration of dissolved oxygen of water is in a range of 6.0-8.5 mg/L, water pH is in a range of 7.0-7.6; environmental parameters in culture ponds of carp parent fish are as follows: photoperiod is in a range of 9-12 h, water temperature is in a range of 16-25° C., water flow velocity is in a range of 25-50 m/min, water transparency is in a range of 90-95 cm, the concentration of dissolved oxygen of water is in a range of 6.5-8.5 mg/L, water pH is in a range of 6.8-7.2; during the parent fish culture of S. grahami and carp, feeding pellet feed twice a day; collecting sperm and eggs from male and female S. grahami parent fish and carp parent fish with mature gonads respectively, artificially inseminating and incubating the eggs from female S. grahami parent fish and carp parent fish; after feeding an incubated fry, obtaining a hybrid variety of S. grahami and carp.

2. The hybridization method of S. grahami and carp according to claim 1 , wherein the environmental parameters of parent fish culture of S. grahami are rhythmically regulated: in Stage 1: water temperature is 15-20° C., water flow velocity is 10-20 cm/s, dissolved oxygen of water is 7.0-8.0 mg/L, and water pH is 7.0-7.6; in Stage 2: water temperature is in a range of 18-22° C., water flow velocity is in a range of 5-15 cm/s, the concentration of dissolved oxygen of water is in a range of 7.0-8.5 mg/L, and water pH is in a range of 7.2-7.4; in Stage 3: water temperature is in a range of 15-20° C., water flow velocity is in a range of 5-15 cm/s, the concentration of dissolved oxygen of water is in a range of 6.0-7.0 mg/L, and water pH is in a range of 7.0-7.6.

3. The hybridization method of S. grahami and carp according to claim 1 , wherein the environmental parameters of parent fish culture of carp are rhythmically regulated: in Stage 1′: photoperiod is in a range of 9-10 h, water temperature is in a range of 16-20° C., water flow velocity is in a range of 30-50 m/min, water transparency is in a range of 90-95 cm, the concentration of dissolved oxygen of water is in a range of 6.5-7.5 mg/L, and water pH is in a range of 6.8-7.2; in Stage 2′: photoperiod is in a range of 11-12 h, water temperature is in a range of 18-25° C., water flow velocity is in a range of 25-40 m/min, water transparency is in a range of 90-95 cm, the concentration of dissolved oxygen of water is in a range of 7.0-8.5 mg/L, and water pH is in a range of 7.0-7.2; in Stage 3′: photoperiod is in a range of 11-12 h, water temperature is in a range of 18-25° C., water flow velocity is in a range of 40-45 m/min, water transparency is in a range of 90-95 cm, the concentration of dissolved oxygen of water is in a range of 7.0-7.5 mg/L, and water pH is in a range of 6.8-7.2.

4. The hybridization method of S. grahami and carp according to claim 1 , wherein during the parent fish culture of S. grahami and carp, the pellet feed is rhythmically regulated according to requirements: feeding feed SGF1 in Stage 1 or Stage 1′, wherein the feed SGF1 is a mixture of mash feed containing 30% crude protein, shrimp, vitamin D3 calcium tablets, multi-vitamin tablets and water, with a mass ratio of 250:20:10:8:300; feeding feed SGF2 in Stage 2 or Stage 2′, wherein the feed SGF2 is a mixture of mash feed containing 35% crude protein, vitamin D3 calcium tablets, multi-vitamin tablets and water, with a mass ratio of 280:20:10:300; feeding feed SGF3 in Stage 3 or Stage 3′, wherein the feed SGF3 is a mixture of mash feed containing 32% crude protein, vitamin D3 calcium tablets, vitamin C and water, with a mass ratio of 250:20:20:300.

5. The hybridization method of S. grahami and carp according to claim 1 , wherein the artificially inseminating comprises using S. grahami as female parent and carp as male parent, and using S. grahami as male parent and carp as female parent.

6. The hybridization method of S. grahami and carp according to claim 1 , wherein the quantity ratio of eggs to sperm is 1-3:1-10×10 5 during the artificial inseminating.

7. The hybridization method of S. grahami and carp according to claim 1 , wherein the fry is fed with feed 7 days after the fry breaks an egg membrane.

8. The hybridization method of S. grahami and carp according to claim 7 , wherein the feeding of the feed is adjusted according to different growth periods of fry: feeding pulpous feed SCF1 from the 1st day to the 7th day of feeding, 4 times per day with the feeding amount of 8-10 mL/m 2 , wherein the pulpous feed SCF1 is a mixture of boiled egg yolk, shrimp, multi-vitamin tablets and water, with a mass ratio of 300:20:10:750; feeding feed SCF2 and rotifer from the 8th to 20th day, 3 times per day, the feeding amount of rotifer each time is 1.5 million/m 2 , wherein the feed SCF2 is a mixture of boiled egg yolk, shrimp, freshly ground soybean milk, multi-vitamin tablets and water, with a mass ratio of 250:20:250:10:750; feeding feed SCF3 from the 21st to 60th day, twice a day, the feeding amount is 10-12 g/m 2 , wherein the feed SCF3 is a mixture of freshly ground soybean milk, shrimp, mash feed containing 28% crude protein, vitamin D3 calcium tablets, multi-vitamin tablets and water, with a mass ratio of 250:10:250:10:750; after feeding for 60 days, feeding the pellet feed used for parent fish culture, and the feeding amount is according to the feed coefficient of 3%-4%, and the feed is consumed within 15-30 min.

9. The hybridization method of S. grahami and carp according to claim 8 , wherein the water quality is regulated by adding nitrifying bacteria in an amount of 15-17 g/m 2 every day during the fry rearing period.

10. The hybridization method of S. grahami and carp according to claim 8 , wherein each feeding amount of the pellet feed is according to the feed coefficient of 3%-4%; the pellet feed is a mixture of mash feed containing 30%-35% crude protein, vitamin D3 calcium tablets, multi-vitamin tablets and water, with a mass ratio of 250-280:10-20:8-20:300; after artificial inseminating, rinsing the sperm-egg mixture with normal saline with osmotic pressure of 280-320 mOsm/L and temperature of 18-22° C. for 3-4 times, then splashing the sperm-egg mixture on a palmsheet, and incubating the palmsheet adhered with fertilized eggs in incubating pond; the incubating parameters of the incubating pond are as follows: water level is 1.2 m, water surface is in a range of 40-80 cm above fertilized eggs, no direct light source, water temperature is in a range of 18-22° C., water pH is in a range of 6.8-7.5, the concentration of dissolved oxygen of water is in a range of 7.0-8.0 mg/L; during incubation, sterilizing the fertilized eggs in potassium permanganate solution for 15 min every day for 5 days, the fry is incubated on the 6th day and the incubation ends on the 9th day.

11. The hybridization method of S. grahami and carp according to claim 6 , wherein the artificially inseminating comprises using S. grahami as female parent and carp as male parent, and using S. grahami as male parent and carp as female parent.

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