A family breeding method for large-scale barbel

Abstract

The invention provides a family breeding method for large-scale barbel, comprising the following steps: one, selecting parents; two, disinfecting; three, cultivating parents; four, artificially inducing spawning; five, artificial insemination; six, cultivating fry; and seven, late family cultivation. The invention introduces high-quality exogenous genes of wild large-scale barbel, which can effectively improve the body color differentiation, frequent diseases, slow growth, decreased fecundity and other phenomena of existing cultured large-scale barbel, and is helpful to breed new strains with excellent growth traits such as fast growth. The invention significantly improves the fertilization rate, hatching rate and fry survival rate of large-scale barbel by optimizing conditions such as cultivating parent fish, types and concentration ratios of oxytocin, injection methods and feed types.

Description

Technical Field

[0001] The invention relates to the technical field of large-scale barbel breeding, and in particular to a family selection and breeding method for large-scale barbel. Background Art

[0002] The large-scale barb belongs to the Cyprinidae, the subfamily Barbinae, and the genus Barb. It is an upstream migratory fish and an endangered and valuable large economic fish. The large-scale barb has a variety of diets, delicious meat, fast growth, strong resistance to adversity, and salt and alkali tolerance. It is a wide-temperature (survival water temperature is 1-30℃) and moderately salt-alkali tolerant fish.

[0003] As an alien aquaculture species, the bigscale barbel has a small parental population. Inbreeding depression and decreased genetic diversity caused by the founder effect gradually appear in individuals, leading to a decline in seed quality and causing problems such as decreased growth rate and frequent diseases in aquaculture production. Therefore, genetic improvement of the bigscale barbel and breeding of new strains with excellent growth traits such as rapid growth are necessary conditions to ensure the sustained, healthy and stable development of the bigscale barbel industry.

[0004] Selective breeding is a technical method that uses existing varieties to select natural variation groups that meet the needs of production and life through artificial screening, and obtains new varieties with excellent traits, stable inheritance, and that meet the selection requirements after multiple generations of screening. Among them, family selection is one of the important methods in traditional selective breeding methods. It refers to selective breeding based on families, and compares and analyzes the growth traits of each family. The average value of the target trait is used as a parameter, and families with high average values ​​are retained, and families with low average values ​​are eliminated, thereby improving the genetic homozygosity of the varieties from generation to generation, and then breeding new varieties with stable and excellent traits.

[0005] However, since the screening of excellent pedigrees of large scale barbel is still in a blank stage, whether it is possible to screen large scale barbel fry with excellent traits and rapid growth has become one of the main directions at present. Therefore, providing a pedigree breeding method for large scale barbel has become an urgent problem to be solved. Summary of the invention

[0006] In order to solve the above problems, the present invention provides a family breeding method for large-scale barbel, which specifically comprises the following steps:

[0007] Step 1: Select parents

[0008] Selecting 4-5 year old wild male barbels with no parasites, strong physique, no external injuries, intact scales, strong vitality and mature gonads from natural water bodies, and selecting female barbels with a body length of ≥40 cm and a weight of ≥850 g from aquaculture water bodies, and using the male and female fish as parents for breeding;

[0009] Wild fish live in the natural environment and follow the cruel laws of nature. Those that survive are individuals with excellent traits. In the process of facing natural enemies and coexisting with microorganisms, wild fish make their meat firmer, more resistant to diseases, and have more genetic advantages.

[0010] Step 2: Disinfection

[0011] Use chlorine dioxide effervescent tablets to disinfect the water in the culture pool, with a dosage of 10-15g / m 3 , use it 2-5 times continuously, with an interval of 10-20 minutes each time; before placing the parent in the cultivation pond, soak it in a potassium permanganate solution with a mass fraction of 1-3% for 10-30 minutes; when the parent is placed in the cultivation pond, the density is 1-1.5 tails / m 3 , the male-female ratio is (1-2):(1-3);

[0012] Using chlorine dioxide to disinfect the breeding pond can kill bacteria in the fish tank, reduce the chance of fish getting sick, and eliminate the fishy smell and odor in the pond; using quicklime and bleaching powder to disinfect the fish pond requires controlling the dosage. Once the dosage is too large, it will cause irreversible damage to the fish body. Chlorine dioxide has almost no negative impact on the fish body and does not require precise dosage control. In addition, chlorine dioxide can appropriately increase the oxygen content in the water.

[0013] Step 3: Cultivate the parents

[0014] Feed 2-3 times a day, each feeding amount is 4-5% of the total weight of the fish; water quality conditions: dissolved oxygen at the bottom of the pond ≥ 6mg / L, ammonia nitrogen ≤ 0.01mg / L, nitrite ≤ 0.01mg / L, water transparency 15-20cm, water temperature 23-27℃, pH 7-8; change water every 5-7d, and the amount of water changed is ≥ 1 / 3 of the total water volume; add chlorine dioxide effervescent tablets every 10-15d, with a dosage of 10-15g / m 3 ; Antimicrobial drugs were administered every 15 days;

[0015] Preferably, the ingredients of the feed are dried shrimp, bean curd dregs, fish meal, wheat bran, fish oil, rapeseed cake, and peanut powder, with a mass ratio of (4-8):(3-7):(2-6):(5-7):(7-9):(3-5):(4-5);

[0016] The protein content required by large-scale barbel is relatively high, especially when producing a large number of reproductive cells, it needs to consume more protein. Shrimp skin, tofu dregs, and fish meal contain a lot of high-quality protein; fat is an important nutrient necessary for the growth of large-scale barbel, which can provide energy for its life activities. Fish oil and peanut powder can provide a lot of high-quality fat; adding an appropriate amount of carbohydrates, such as wheat bran, to barbel feed can enhance its physique and promote its growth; rapeseed cake contains a large amount of vitamins and trace mineral elements, which can increase the growth rate and weight of large-scale barbel, and is beneficial to enhance its immunity and resist the invasion of bacteria, and plays a vital role in regulating the metabolism of large-scale barbel;

[0017] Preferably, the antibacterial drug is allicin, and the dosage is 10-20 mg / tail;

[0018] Allicin has a strong bactericidal and anti-inflammatory effect. It can inhibit a variety of Gram-positive cocci and Gram-negative bacilli at extremely low concentrations. It also has inhibitory effects on molds, viruses, protozoa, pinworms, etc. Allicin is environmentally friendly and will not pollute water quality. It is a safe and reliable green antibacterial drug.

[0019] Step 4: Artificial induction of labor

[0020] After the parents have been cultivated for 1-2 months, the induced spawning pool is disinfected in the same way as in step 2. The parents are then transferred to the induced spawning pool and the males and females are separated. The water temperature is 28±1℃, and the parents are then marked with PIT.

[0021] The amount of feed is the same as step 3. On this basis, the induced labor drug is fed once every 2 days, the bloodworm is fed once a day, and the water flushing stimulation is performed once a day. After 7 days, the parents are injected with induced labor drugs once, and artificial insemination is performed 12 hours after the injection.

[0022] The large bloodworm can promote estrus in fish; feeding oxytocin drugs can make fish slowly absorb oxytocin hormones, gradually increase the number of reproductive cells in the fish, and make the fish gradually adapt to the high-concentration hormone environment. Then injecting oxytocin drugs can greatly improve the production performance of fish.

[0023] Preferably, the oxytocin drug is gonadotropin, and the feeding amount is 1.5-2.5 mg / tail;

[0024] Preferably, the feeding amount of the large bloodworm is 10-30g / tail;

[0025] Preferably, the female fish is injected with 2-3 mg / tail of the oxytocin agent, and the male fish is injected with 1 / 2-1 / 3 of the female fish, and the composition of the oxytocin agent is PG+HCG, and the mass ratio is (1-3):(1-3);

[0026] Step 5: Artificial insemination

[0027] Squeeze the abdomen of the female fish to squeeze the female fish eggs into a container, squeeze the male fish semen into the container in the same way, add 100-200 ml of physiological saline with a mass concentration of 0.8-0.9%, then mix the fish eggs with the diluted semen, stir for 30-60 seconds, and then transfer to an incubation facility for incubation. The fish eggs of the same family are placed in the same incubation facility;

[0028] Preferably, the incubation facility is an incubation bucket, the water temperature in the incubation bucket is 18-26°C, the incubation time is 50-80h, and the pH is 6.5-7.5;

[0029] Step 6: Fry cultivation

[0030] After the fry are hatched for 2-3 days, feed the fry feed at 1-3% of the total weight of the fry every day, divided into three feedings, and the mass ratio of each feeding amount is 1:2:1;

[0031] After 14 days of hatching, feed the fry with fry feed and live feed at a rate of 2-3% of the total weight of the fry every day, divided into three feedings, with the mass ratio of each feeding being 3:4:3;

[0032] After the fry are hatched for 30 days, feed 3-4% of the total weight of the fry is fed every day, and the feeding is carried out three times, and the mass ratio of each feeding amount is 1:1:1, and the feed composition is consistent with the feed composition in step 3;

[0033] After 45 days of hatching, the fry are fed 2-3 times a day, with the amount of each feed being 3-4% of the total weight of the fish. The feed ingredients are consistent with those in step 3.

[0034] During the growth of fry, gradually adding live feed to the fry nutrient feed can gradually improve the fry's adaptability to the environment, increase the fry's resistance to stress and disease, and make the fry stronger.

[0035] Preferably, the method for preparing the fry feed is to grind tofu, boiled eggs, cabbage and carrots in a mass ratio of 3:5:2:3 and mix them evenly;

[0036] Preferably, the mass ratio of the fry feed to the live feed is (8-10):(1-2);

[0037] Preferably, the live feed is Paramecium and rotifer, with a mass ratio of 1:3;

[0038] Step 7: Family Cultivation

[0039] When large-scale barbel families are selected and grown to 60 days, 90 days, 120 days and 240 days of age, 30-50 fish are randomly sampled from each family for measurement of body length, weight and excellent traits. Excellent large-scale barbel families are screened out and cultivated as the next generation of parents, and then a new round of family selection and breeding is carried out.

[0040] The present invention has the following advantages:

[0041] (1) The invention introduces high-quality exogenous genes from wild barbel, which can effectively improve the body color differentiation, frequent diseases, slow growth, and decreased reproductive capacity of existing cultured barbel, and is helpful for breeding new strains with excellent growth traits such as fast growth;

[0042] (2) The present invention significantly improves the fertilization rate, hatching rate and fry survival rate of large-scale barbel by optimizing the cultivation of broodstock, the type and concentration ratio of oxytocin, the injection method and the type of feed. DETAILED DESCRIPTION

[0043] The technical solutions in the embodiments of the invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the invention, not all of the embodiments. Based on the embodiments of the invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the invention.

[0044] Example 1

[0045] Step 1: Select parents

[0046] Selecting 4-5 year old wild male barbels with no parasites, strong physique, no external injuries, intact scales, strong vitality and mature gonads from natural water bodies, and selecting female barbels with a body length of ≥40 cm and a weight of ≥850 g from aquaculture water bodies, and using the male and female fish as parents for breeding;

[0047] Step 2: Disinfection

[0048] Use chlorine dioxide effervescent tablets to disinfect the water in the culture pool, with a dosage of 15g / m 3 , used three times in succession, with an interval of 15 minutes between each use; before the parent is placed in the culture pond, it is soaked in a 3% potassium permanganate solution for 20 minutes; when the parent is placed in the culture pond, the density is 1 tail / m 3 , the male-female ratio is 1:1.5, including 100 males and 150 females;

[0049] Step 3: Cultivate the parents

[0050] Feed was given three times a day, and the amount of each feed was 4% of the total weight of the fish; the ingredients of the feed were dried shrimp, bean curd residue, fish meal, wheat bran, fish oil, rapeseed cake, and peanut powder, and the mass ratio was (4-8):(3-7):(2-6):(5-7):(7-9):(3-5):(4-5); water quality conditions: dissolved oxygen at the bottom of the pond was 6 mg / L, ammonia nitrogen content was 0.005 mg / L, nitrite content was 0.005 mg / L, pool water transparency was 20 cm, water temperature was 25°C, pH 7; water was changed every 5 days, and the water change amount was 1 / 2 of the total water volume; chlorine dioxide effervescent tablets were added every 15 days, and the dosage was 10 g / m 3 ; Anti-allicin is administered once every 15 days, with the dosage of 10-20 mg / tail;

[0051] Step 4: Artificial induction of labor

[0052] After the parents were cultivated for 1.5 months, the induced spawning pool was disinfected in the same way as in step 2. The parents were then transferred to the induced spawning pool and the males and females were separated. The water temperature was 28 ± 1 °C and the parents were then marked with PIT.

[0053] The feeding amount is the same as step 3. On this basis, gonadotropin is fed once every 2 days at a feeding amount of 2 mg / tail; large bloodworms are fed once a day at a feeding amount of 20 g / tail, and water stimulation is performed once a day. After 7 days, female fish are injected with PG1mg+HCG 2mg, and male fish are injected with 1 / 2 of the female fish. Artificial insemination is performed 12 hours after the injection;

[0054] Step 5: Artificial insemination

[0055] Squeeze the abdomen of the female fish and squeeze the female fish eggs into the container. Use the same method to squeeze the male fish semen into the container, and add 200ml of 0.9% saline solution. Then mix the fish eggs with the diluted semen, stir for 30s, and then transfer them to the hatching bucket for hatching. The eggs of the same family are placed in the same hatching bucket. The water temperature is 20℃, the incubation time is 72h, and the pH is 7.

[0056] Step 6: Fry cultivation

[0057] After the fry are hatched for 2 days, feed the fry feed at 1% of the total weight of the fry every day, divided into three feedings, and the mass ratio of each feeding amount is 1:2:1;

[0058] The method for preparing the fry feed is as follows: tofu, boiled eggs, cabbage and carrots in a mass ratio of 3:5:2:3 are crushed and evenly mixed;

[0059] After 14 days of hatching, the fry were fed with fry feed and live feed at a rate of 2% of the total weight of the fry every day, divided into three feedings, with the mass ratio of each feeding amount being 3:4:3; the mass ratio of fry feed to live feed was 8:1;

[0060] The live feed is Paramecium and rotifer, with a mass ratio of 1:3;

[0061] After the fry are hatched for 30 days, feed 3% of the total weight of the fry is fed every day, and the feeding is carried out three times, and the mass ratio of each feeding amount is 1:1:1, and the feed composition is consistent with the feed composition in step 3;

[0062] After 45 days of hatching, the fry were fed twice a day, with the amount of each feeding being 4% of the total weight of the fish, and the feed composition was consistent with the feed composition in step 3;

[0063] Step 7: Family Cultivation

[0064] When the large-scale barbel families were selected and grown to 60 days, 90 days, 120 days and 240 days of age, 30-50 fish were randomly sampled from each family for measurement of body length, weight and excellent traits. The results are shown in Table 1. The excellent large-scale barbel families were screened out and cultivated as the next generation of parents, and then a new round of family selection and breeding was carried out.

[0065] Comparative Example 1

[0066] Different from Implementation 1, male large scale barbel in aquaculture water was selected in this comparative example.

[0067] Comparative Example 2

[0068] Different from implementation 1, this comparative ratio did not feed big bloodworms.

[0069] Comparative Example 3

[0070] Different from Implementation 1, in this comparative example, no labor-inducing drug was fed, and the labor-inducing agent was directly injected.

[0071] Comparative Example 4

[0072] Different from implementation 1, this comparative example replaces the fry feed with cooked egg yolk.

[0073] Comparative Example 5

[0074] Different from implementation 1, this comparative example does not feed live bait.

[0075] Test example

[0076] During the process of artificial induction of spawning and artificial insemination, the fertilization rate, hatching rate and survival rate of fish eggs were statistically analyzed, and the results are shown in Table 1;

[0077] Table 1

[0078]

[0079] Note: Fertilization rate = number of fertilized eggs / total number of eggs; hatching rate = total number of fry / number of fertilized eggs; survival rate = number of live fry after 30 days of hatching / total number of fry; body length and weight are average values;

[0080] As shown in Table 1, the large-scale barbel cultured in Example 1 has extremely high fertilization rate, hatching rate and survival rate, and the fry grows rapidly and has strong stress resistance; Comparative Example 1 does not introduce high-quality genes of wild large-scale barbel, which has a significant effect on fertilization rate, hatching rate, survival rate and growth rate; Comparative Examples 2 and 3 use different induced production methods, which have a significant effect on fertilization rate, hatching rate and survival rate, but have no significant effect on fry growth rate; Comparative Examples 4 and 5 use different feeds to feed fry, which have a significant effect on fry growth rate, and the fry have low stress resistance, and diseases occur, but have no significant effect on fertilization rate, hatching rate and survival rate;

[0081] In addition, the fry of Example 1 has a strong physique, firm and elastic meat, bright and shiny body surface, strong vitality, and no obvious fishy smell after cooking in clear water after maturity; the fry of Comparative Example 1 has soft and inelastic meat, dull body surface, low vitality, and obvious fishy smell after cooking in clear water after maturity.

[0082] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for family selection of large-scale barbel, characterized in that: The following steps are involved: Step 1: Select parents Selecting 4-5 year old wild male barbels with no parasites, strong physique, no external injuries, intact scales, strong vitality and mature gonads from natural water bodies, and selecting female barbels with a body length of ≥40 cm and a weight of ≥850 g from aquaculture water bodies, and using the above wild male barbels and female barbels as parents for breeding; Step 2: Disinfection Use chlorine dioxide effervescent tablets to disinfect the water in the culture pool, with a dosage of 10-15g / m 3 , use it 2-5 times continuously, with an interval of 10-20 minutes each time; before placing the parent in the cultivation pond, soak it in a potassium permanganate solution with a mass fraction of 1-3% for 10-30 minutes; when the parent is placed in the cultivation pond, the density is 1-1.5 tails / m 3 , the male-female ratio is (1-2):(1-3); Step 3: Cultivate the parents Feed 2-3 times a day, each feeding amount is 4-5% of the total weight of the fish; water quality conditions: dissolved oxygen at the bottom of the pond ≥ 6mg / L, ammonia nitrogen ≤ 0.01mg / L, nitrite ≤ 0.01mg / L, water transparency 15-20cm, water temperature 23-27℃, pH 7-8; change water every 5-7d, and the amount of water changed is ≥ 1 / 3 of the total water volume; add chlorine dioxide effervescent tablets every 10-15d, with a dosage of 10-15g / m 3 ; Antimicrobial drugs were administered every 15 days; Step 4: Artificial induction of labor After the parents are cultivated for 1-2 months, the induced labor pool is disinfected in the same way as in step 2. Then the parents are transferred to the induced labor pool and the male and female are separated. The water temperature is 28±1℃ and the parents are marked with PIT. The amount of feed is the same as in step 3. On this basis, induced labor drugs are fed once every 2 days, large bloodworms are fed once a day, and flushing stimulation is performed once a day. After 7 days, the parents are injected with induced labor drugs and artificial insemination is performed 12 hours after the injection. Step 5: Artificial insemination Squeeze the abdomen of the female fish to squeeze the female fish eggs into a container, squeeze the male fish semen into the container in the same way, add 100-200 ml of physiological saline with a mass concentration of 0.8-0.9%, then mix the fish eggs with the diluted semen, stir for 30-60 seconds, and then transfer to an incubation facility for incubation. The fish eggs of the same family are placed in the same incubation facility; Step 6: Fry cultivation After the fry are hatched for 2-3 days, feed the fry feed at 1-3% of the total weight of the fry every day, divided into three feedings, and the mass ratio of each feeding amount is 1:2:1; After 14 days of hatching, feed the fry with fry feed and live feed at a rate of 2-3% of the total weight of the fry every day, divided into three feedings, with the mass ratio of each feeding being 3:4:3; After the fry are hatched for 30 days, feed 3-4% of the total weight of the fry is fed every day, and the feeding is carried out three times, and the mass ratio of each feeding amount is 1:1:1, and the feed composition is consistent with the feed composition in step 3; After 45 days of hatching, the fry are fed 2-3 times a day, with the amount of each feed being 3-4% of the total weight of the fish. The feed ingredients are consistent with those in step 3. Step 7: Family Cultivation When large-scale barbel families are selected and grown to 60 days, 90 days, 120 days and 240 days of age, 30-50 fish are randomly sampled from each family for measurement of body length, weight and excellent traits. Excellent large-scale barbel families are screened out and cultivated as the next generation of parents, and then a new round of family selection and breeding is carried out.

2. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step three, the ingredients of the feed are dried shrimp, bean curd dregs, fish meal, wheat bran, fish oil, rapeseed cake, and peanut powder, with a mass ratio of (4-8):(3-7):(2-6):(5-7):(7-9):(3-5):(4-5).

3. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step three, the antibacterial drug is allicin, and the dosage is 10-20 mg / tail.

4. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step 4, the oxytocin drug is gonadotropin, and the feeding amount is 1.5-2.5 mg / tail.

5. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step 4, the feeding amount of the large bloodworm is 10-30g / tail.

6. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step 4, 2-3 mg / tail of oxytocin is injected into female fish, and the injection amount of male fish is 1 / 2-1 / 3 of that of female fish. The composition of the oxytocin is PG+HCG, and the mass ratio is (1-3):(1-3).

7. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step 5, the incubation facility is an incubation bucket, the water temperature in the incubation bucket is 18-26° C., the incubation time is 50-80 hours, and the pH is 6.5-7.

5.

8. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step six, the method for preparing the fry feed is to grind tofu, boiled eggs, cabbage and carrots in a mass ratio of 3:5:2:3 and then evenly mix them.

9. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step six, the mass ratio of the fry feed to the live feed is (8-10):(1-2).

10. The method for family selection of large-scale barbel according to claim 1, characterized in that: In step six, the live feed is Paramecium and rotifer, with a mass ratio of 1:3.

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