CN109430117B - Method for improving reproductive capacity of sepia lycidas - Google Patents
Method for improving reproductive capacity of sepia lycidas Download PDFInfo
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- CN109430117B CN109430117B CN201810978633.8A CN201810978633A CN109430117B CN 109430117 B CN109430117 B CN 109430117B CN 201810978633 A CN201810978633 A CN 201810978633A CN 109430117 B CN109430117 B CN 109430117B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A23K10/00—Animal feeding-stuffs
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- A—HUMAN NECESSITIES
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- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
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- A23K20/158—Fatty acids; Fats; Products containing oils or fats
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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Abstract
The invention discloses a method for improving the fertility of a sepia mimosoides, which comprises the steps of temporarily culturing the prepared sepia mimosoides in an indoor culture pond provided with an egg-attached base, carrying out flowing water stimulation, feeding bait containing fermented soybean meal, collecting fertilized eggs for artificial incubation after the parents of the sepia mimosoides naturally lay eggs, then culturing incubated seedlings in a seedling pond, culturing the seedlings in the culture pond after the parents of the sepia mimosoides grow well, and fermenting the fermented soybean meal by using lactobacillus rhamnosus containing tocopherol and cysteine. The beneficial effects are that: the method can improve the ingestion rate, the utilization rate and the organism immunity of the sepia pseudoacacia parents, can ensure that fertilized eggs are adhered together and are not easy to be dispersed by water flow, improve the bacteriostatic ability of the fertilized eggs, the quality and the hatching rate of the fertilized eggs, inhibit the growth of bacteria and finally improve the reproductive capacity of the sepia pseudoacacia.
Description
Technical Field
The invention relates to the technical field of aquaculture, in particular to a method for improving the fertility of sepia pseudobulb.
Background
The cuttlefish belongs to the phylum mollusca, class cephalopoda, order of ten wrists and order of cuttlefish, is one of the most developed potential types of cephalopoda, has delicious meat quality, and accounts for more than 90 percent of edible parts; its gonad can be processed into "fish ear" and "fish egg"; in addition, the cuttlebone and the ink sac have the effects of blood coagulation, tumor resistance, cancer resistance and the like. Therefore, the cuttlefish is popular with the public and is an important marine fishery resource in China. In recent years, the distribution of cuttlefish resources has been deeply studied at home and abroad, main species which are caught, developed and utilized at present are available, and the yield of sepia esculenta, sepiella maindroni, cuttlefish, sepiella mimosoides, tiger spot cuttlefish and the like is high in China. However, due to over-fishing, deterioration of sea environment and the like, the cuttlefish resources are obviously declined, and fishing flood cannot be formed. The breeding period is an important stage of the life cycle of the cuttlefish, most of the cuttlefish except Australian giant cuttlefish hardly gather in the breeding period, but the cuttlefish has the phenomenon of reproductive migration, and the common cuttlefish has only one breeding period, such as the golden cuttlefish and the Sepiella maindroni in the natural sea area. However, under the condition of artificial cultivation, the sepiella maindroni can breed for 2 times due to the change of environmental factors. The number of brood of parents determines the number and quality of offspring, and limits the number, quality and diversity level of the supplemented population. The ovum amount of different cuttlefish has obvious difference, for example, the ovum amount of Sepiella maindroni is generally 3000 in 1000-. In order to recover, develop and utilize the cuttlefish resource, foreign scholars have earlier developed the research on artificial breeding of cuttlefish. The research on the breeding and breeding technology of the sepia esculenta in China mainly focuses on the sepia maindroni, sepia esculenta and sepia tiger, and the research on the breeding and breeding technology of the sepia mimosoides is less.
The sepia pseudoacacia enters the breeding period and has obvious puppet behavior, and males often fight for puppets; when the squid is used for puppet, the squid does not eat food. After successful mating, males are accompanied by guarding behavior to avoid the interference of females with other males, which may be a mating strategy to increase fertility and hatchability. During mating, a male uses a stem-like wrist to convey a spermatophore into a female seminal vesicle, and after fertilization, the female individual selects sessile algae, branches, shells and the like as an attachment base to lay eggs. The sepia peltata exhibits collective oviposition, is bonded on the same egg chain to form a grape-shaped public oviposition area, has average oviposition amount of 354 grains, is similar to sepia maindroni, does not have obvious egg protecting action, and excessively catches young sepia peltata, overwintering sepia and oviposition parents in a breeding period, so that resource supplement groups of the sepia peltata receive serious threat and resource decline is caused. Therefore, the method for researching how to improve the fertility of the sepia pseudobulb can enrich the biological knowledge of the sepia pseudobulb, provide theoretical basis for further developing artificial breeding of the sepia pseudobulb on a large scale, and have important significance for improving the artificial breeding technology, promoting large-scale breeding and recovering population resources.
Disclosure of Invention
The invention aims to provide a method for improving the fecundity of sepiella squids, which can improve the ingestion rate, the utilization rate and the organism immunity of the parents of the sepiella squids, can ensure that fertilized eggs are adhered together and are not easy to be dispersed by water flow, improve the bacteriostatic ability of the fertilized eggs, the quality and the hatchability of the fertilized eggs, inhibit the growth of bacteria and improve the reproductive capacity of the sepiella squids.
Aiming at the problems mentioned in the technology, the invention adopts the technical scheme that:
a method for improving the fertility of sepia pseudoacacia includes such steps as temporarily culturing the sepia pseudoacacia in an indoor culture pool with egg-attached base, stimulating by flowing water, feeding the bait containing fermented soybean dregs, collecting fertilized eggs, artificial incubation, culturing the incubated seedlings in seedling pool, culturing in culture pool, and fermenting the fermented soybean dregs with rhamnose lactobacillus containing tocopherol and cysteine. The method can improve the ingestion rate, the utilization rate and the body immunity of the sepia pseudoacacia parents, improve the expression of the factor genes participating in the promotion of oocyte maturation, reduce the expression of the factor inhibiting oocyte maturation, promote the maturation of the oocytes, increase the egg laying amount, the fertilization rate and the hatching rate, and further improve the fertility of the sepia pseudoacacia; the fertilized eggs can be adhered together and are not easy to be dispersed by water flow, and the bacteriostatic ability, the fertilized egg quality and the hatching rate of the fertilized eggs are improved; the used egg-attaching base has good flexibility, antibacterial property, adhesive property and affinity property, can effectively inhibit the growth of bacteria, avoids the sepia pseudosciaenae from being injured and influencing the egg laying, enables fertilized eggs to be firmly attached to the egg-attaching base, improves the egg quality of the fertilized eggs, further improves the hatchability and finally improves the reproductive capacity of the sepia pseudosciaenae.
Preferably, the egg-attached base is a polyethylene mesh sheet with a mesh size of 25-30 mm. This attach ovum base can not cause the cuttlefish injury and influence to lay eggs, but reuse, resistant long-time soaking in the sea water, do not spoil quality of water, it is good to attach the ovum effect, can shorten the spawning time, 1min after the male and female mating, female just begins to attach the ovum on attaching the ovum base, it is even firm to adhere to, improve the egg production, it can reach 809 pieces/piece at most to attach the ovum volume, can also avoid the fertilized ovum of cuttlefish to milden and rot or the necrosis, improve the ovum quality, and then improve the hatchability, finally improve the cuttlefish reproductive capacity.
Further preferably, the preparation method of the egg-attached base comprises the following steps: fully mixing polyethylene and epsilon-polylysine with the weight ratio of 100:0.1-3 on a double-rod mixing mill at 163 ℃ under 158-. The polyethylene mesh-attached egg base has the functions of polyethylene and epsilon-polylysine, has excellent antibacterial effect, can effectively inhibit the growth of bacteria, can exert good antibacterial capability for a long time, is not easily polluted by the excrement or ink of the cuttlefish, avoids the mouldness or necrosis of fertilized eggs of the cuttlefish, improves the egg quality of the fertilized eggs, assists in improving the reproductive capacity of the cuttlefish, and simultaneously, the addition of the epsilon-polylysine does not influence the mechanical property of the polyethylene mesh and has no adverse influence on the fertilized eggs of the cuttlefish.
More preferably, epsilon-polylysine contains 0.03 to 0.06 wt% of D-lysine. The existence of D-lysine in the reaction system can fully disperse epsilon-polylysine in polyethylene, enhance the compatibility and blending property of the epsilon-polylysine and the polyethylene, improve the service performance and biocompatibility of the egg-attached matrix, has no adverse effect on fertilized eggs of the sepia esculenta, can assist in improving the reproductive capacity of the sepia esculenta, can be matched with epsilon-polylysine, weakens the acting force between polyethylene, hinders the formation of a crystal structure, reduces the crystallinity of the polyethylene, so that the egg-attached base has good flexibility, avoids the sepia pseudosciaenae from being injured and influencing the egg laying, in addition, the addition of the D-lysine is beneficial to improving the cell adhesion performance and affinity performance of the egg-attached base, so that the fertilized eggs can be firmly attached to the egg-attaching base, thereby improving the hatching rate and finally improving the reproductive capacity of the sepia pseudosciaenae.
Preferably, the specific fermentation steps for fermenting the soybean meal are as follows: grinding the soybean meal to 80-120 meshes, adding water until the water content is 45-53%, uniformly mixing, then sterilizing at high pressure for 20-30min, cooling, inoculating lactobacillus rhamnosus with the inoculation amount of 3-5%, fermenting for 1-2d at 35-40 ℃, freeze drying, crushing, and sieving with a 200-mesh and 300-mesh sieve to obtain the fermented soybean meal. In addition, lactobacillus rhamnosus in the fermented soybean meal can improve the expression of a factor gene participating in promotion of oocyte maturation and reduce the expression of a factor inhibiting oocyte maturation, promote the maturation of oocytes, increase the egg yield, fertility rate and hatching rate, further improve the fertility of the cuttlefish, and produce enzymes such as amylase, protease and lipase for enhancing the digestion of a host in the intestinal tract of the host to promote the digestion of the host, so as to promote the digestion rate of organic substances and proteins of the host, and the fermented soybean meal can improve the ingestion rate, the utilization rate, the immunity and the fertility of the sepia lycidas.
Preferably, the tocopherol is 3.5-5.6% by weight of the lactobacillus rhamnosus, and the cysteine is 0.8-1.5% by weight of the lactobacillus rhamnosus. The special existence of the tocopherol and the cysteine can provide sufficient carbon source and nitrogen source for the lactobacillus rhamnosus, promote the growth of lactobacillus rhamnosus, prolong the logarithmic phase of the lactobacillus rhamnosus, enable the lactobacillus rhamnosus to metabolize more metabolites such as organic acid, enzymes and extracellular polysaccharide, change the molecular structure in the bean pulp, enable the bean pulp to have more hydrophilic groups and be beneficial to the absorption and utilization of the bean pulp; on the other hand, the composition can be absorbed and utilized by the sepia pseudomanilensis, the content of FMRFamide in gray matter and white matter is improved, then the yield of secretion with the function of adhering egg strings in the ootheca and the paraootheca is increased, fertilized eggs are adhered together and are not easy to be dispersed by water flow, the hatchability is improved, bacteria capable of producing antibiotics in the lumen of the paraootheca of a female parent can be stimulated to propagate in a large quantity, and then the bacteria are attached to the tertiary egg membranes of the eggs along with the secretion, the bacteriostatic ability of the fertilized eggs is improved, and the hatchability of the fertilized eggs is improved; in conclusion, the special existence of the tocopherol and the cysteine can improve the hatching rate of the fertilized eggs of the sepia pseudoacacia.
Preferably, the bait comprises the following components in parts by weight: 18-28 parts of fermented soybean meal, 10-20 parts of fish meal, 10-20 parts of bone meal, 7-15 parts of shrimp bran powder, 0.002-0.004 part of vitamin A, 0.001-0.002 part of vitamin E, 0.002-0.005 part of vitamin C, 3-7 parts of vegetable oil, 0.2-0.4 part of citric acid and 0.1-0.3 part of phytase.
Preferably, the conditions for parent breeding are: the water temperature is 20-25 deg.C, the salinity of seawater is 25-30, the photoperiod L: D is 4:18-25, the flow velocity of flowing water stimulation is 5-8m/min, the time is 10-15min, the bait is thrown for 2-4 times daily, and the total feeding amount is 0.3-0.6 times of the parent weight.
Preferably, the selection and matching of parents specifically comprises: according to the natural breeding method of the sepia pseudosciaenae, healthy and sexually mature male and female sepia pseudosciaenae are respectively selected as parents, and then the male and female sepia pseudosciaenae are matched according to the ratio of 2-4: 1.
Preferably, the artificial hatching is specifically as follows: putting the fertilized eggs into a culture pond, controlling the water temperature of the culture pond to be 20-25 ℃, the salinity of seawater to be 25-30, the water-soluble oxygen content to be more than or equal to 6mg/L, and the incubation time to be 20-35d, and covering a single-layer sunshade net above the incubation pond.
Compared with the prior art, the invention has the advantages that: 1) the method can improve the ingestion rate, the utilization rate and the body immunity of the sepia pseudoacacia parents, improve the expression of the factor genes participating in the promotion of oocyte maturation, reduce the expression of the factor inhibiting oocyte maturation, promote the maturation of the oocytes, increase the egg laying amount, the fertilization rate and the hatching rate, and further improve the fertility of the sepia pseudoacacia; 2) the method can adhere the fertilized eggs together, is not easy to be dispersed by water flow, and improves the bacteriostatic ability, the quality and the hatchability of the fertilized eggs; 3) the egg-attaching base used in the method has good flexibility, antibacterial property, adhesive property and affinity property, can effectively inhibit the growth of bacteria, prevents the sepiella from being injured and influencing spawning, enables fertilized eggs to be firmly attached to the egg-attaching base, improves the egg quality of the fertilized eggs, further improves the hatchability, and finally improves the reproductive capacity of the sepiella.
Drawings
FIG. 1 shows the results of the tests of average egg attachment amount, hatchability and survival rate in example 6.
Detailed Description
The scheme of the invention is further illustrated by the following examples:
example 1:
a method for improving the fertility of sepia pseudoacacia includes such steps as temporarily culturing the sepia pseudoacacia in an indoor culture pool with egg-attached base, stimulating by flowing water, feeding the bait containing fermented soybean dregs, collecting fertilized eggs, artificial incubation, culturing the incubated seedlings in seedling pool, culturing in culture pool, and fermenting the fermented soybean dregs with rhamnose lactobacillus containing tocopherol and cysteine. The method can improve the ingestion rate, the utilization rate and the organism immunity of the sepia pseudoacacia parents, improve the expression of the factor genes participating in the promotion of oocyte maturation, reduce the expression of the factor inhibiting oocyte maturation, promote the maturation of the oocytes, increase the egg laying amount, the fertilization rate and the hatching rate, and further improve the fertility of the sepia pseudoacacia; the fertilized eggs can be adhered together and are not easy to be dispersed by water flow, and the bacteriostatic ability, the fertilized egg quality and the hatching rate of the fertilized eggs are improved; the used egg-attaching base has good flexibility, antibacterial property, adhesive property and affinity property, can effectively inhibit the growth of bacteria, avoids the sepia pseudosciaenae from being injured and influencing the egg laying, enables fertilized eggs to be firmly attached to the egg-attaching base, improves the egg quality of the fertilized eggs, further improves the hatchability and finally improves the reproductive capacity of the sepia pseudosciaenae.
The egg-attached base is a polyethylene mesh sheet with a mesh of 30 mm. This attach ovum base can not cause the cuttlefish injury and influence to lay eggs, but reuse, resistant long-time soaking in the sea water, do not spoil quality of water, it is good to attach the ovum effect, can shorten the spawning time, 1min after the male and female mating, female just begins to attach the ovum on attaching the ovum base, it is even firm to adhere to, improve the egg production, it can reach 809 pieces/piece at most to attach the ovum volume, can also avoid the fertilized ovum of cuttlefish to milden and rot or the necrosis, improve the ovum quality, and then improve the hatchability, finally improve the cuttlefish reproductive capacity.
The preparation method of the adlay base comprises the following steps: fully mixing polyethylene and epsilon-polylysine with the weight ratio of 100:0.1 on a double-roll mixing mill at 163 ℃ and 280rpm for 12min, then putting the mixed composite material into a mould, transferring the mould onto a flat vulcanizing machine, carrying out hot pressing at 158 ℃ and 12MPa for 4min, then carrying out cold pressing at room temperature, and taking out the mould after the cold pressing is finished, thus obtaining the polyethylene mesh sheet attached egg base. The epsilon-polylysine contains 0.06 wt% of D-lysine. The existence of D-lysine in the reaction system can fully disperse epsilon-polylysine in polyethylene, enhance the compatibility and blending property of the epsilon-polylysine and the polyethylene, improve the service performance and biocompatibility of the egg-attached matrix, has no adverse effect on fertilized eggs of the sepia esculenta, can assist in improving the reproductive capacity of the sepia esculenta, can be matched with epsilon-polylysine, weakens the acting force between polyethylene, hinders the formation of a crystal structure, reduces the crystallinity of the polyethylene, so that the egg-attached base has good flexibility, avoids the sepia pseudosciaenae from being injured and influencing the egg laying, in addition, the addition of the D-lysine is beneficial to improving the cell adhesion performance and affinity performance of the egg-attached base, so that the fertilized eggs can be firmly attached to the egg-attaching base, thereby improving the hatching rate and finally improving the reproductive capacity of the sepia pseudosciaenae. The polyethylene mesh-attached egg base has the functions of polyethylene and epsilon-polylysine, has excellent antibacterial effect, can effectively inhibit the growth of bacteria, can exert good antibacterial capability for a long time, is not easily polluted by the excrement or ink of the cuttlefish, avoids the mouldness or necrosis of fertilized eggs of the cuttlefish, improves the egg quality of the fertilized eggs, assists in improving the reproductive capacity of the cuttlefish, and simultaneously, the addition of the epsilon-polylysine does not influence the mechanical property of the polyethylene mesh and has no adverse influence on the fertilized eggs of the cuttlefish.
The specific fermentation steps of the fermented soybean meal are as follows: grinding soybean meal to 80 meshes, adding water until the water content is 53%, uniformly mixing, autoclaving for 20min, cooling, inoculating lactobacillus rhamnosus with an inoculum size of 5%, fermenting at 35 deg.C for 2d, freeze drying, pulverizing, and sieving with 200 meshes sieve to obtain fermented soybean meal. In addition, lactobacillus rhamnosus in the fermented soybean meal can improve the expression of a factor gene participating in promotion of oocyte maturation and reduce the expression of a factor inhibiting oocyte maturation, promote the maturation of oocytes, increase the egg yield, fertility rate and hatching rate, further improve the fertility of the cuttlefish, and produce enzymes such as amylase, protease and lipase for enhancing the digestion of a host in the intestinal tract of the host to promote the digestion of the host, so as to promote the digestion rate of organic substances and proteins of the host, and the fermented soybean meal can improve the ingestion rate, the utilization rate, the immunity and the fertility of the sepia lycidas.
The tocopherol accounts for 5.6 percent of the weight of the lactobacillus rhamnosus, and the cysteine accounts for 0.8 percent of the weight of the lactobacillus rhamnosus. The special existence of the tocopherol and the cysteine can provide sufficient carbon source and nitrogen source for the lactobacillus rhamnosus, promote the growth of lactobacillus rhamnosus, prolong the logarithmic phase of the lactobacillus rhamnosus, enable the lactobacillus rhamnosus to metabolize more metabolites such as organic acid, enzymes and extracellular polysaccharide, change the molecular structure in the bean pulp, enable the bean pulp to have more hydrophilic groups and be beneficial to the absorption and utilization of the bean pulp; on the other hand, the composition can be absorbed and utilized by the sepia pseudomanilensis, the content of FMRFamide in gray matter and white matter is improved, then the yield of secretion with the function of adhering egg strings in the ootheca and the paraootheca is increased, fertilized eggs are adhered together and are not easy to be dispersed by water flow, the hatchability is improved, bacteria capable of producing antibiotics in the lumen of the paraootheca of a female parent can be stimulated to propagate in a large quantity, and then the bacteria are attached to the tertiary egg membranes of the eggs along with the secretion, the bacteriostatic ability of the fertilized eggs is improved, and the hatchability of the fertilized eggs is improved; in conclusion, the special existence of the tocopherol and the cysteine can improve the hatching rate of the fertilized eggs of the sepia pseudoacacia.
The bait comprises the following components in parts by weight: 28 parts of fermented soybean meal, 10 parts of fish meal, 20 parts of bone meal, 7 parts of mysidacea powder, 0.004 part of vitamin A, 0.001 part of vitamin E, 0.005 part of vitamin C, 3 parts of vegetable oil, 0.4 part of citric acid and 0.1 part of phytase.
The conditions for parent breeding were: the water temperature is 25 deg.C, the salinity of seawater is 25, the photoperiod L: D is 4:25, the flow velocity of flowing water stimulation is 5m/min, the time is 15min, the bait is thrown for 2 times a day, and the total feeding amount is 0.6 times of the parent weight.
The parent selection and matching specifically comprises the following steps: according to the natural breeding method of the sepia pseudosciaenae, healthy and sexually mature male and female sepia pseudosciaenae are respectively selected as parents, and then the male and female sepia pseudosciaenae are matched according to the ratio of 2: 1.
The artificial incubation comprises the following specific steps: and (3) placing the fertilized eggs in a culture pond, controlling the water temperature of the culture pond to be 20 ℃, the salinity of seawater to be 30, the water-soluble oxygen content to be more than or equal to 6mg/L and the incubation time to be 20d, and covering a single-layer sunshade net above the incubation pond.
Example 2:
a method for improving the fertility of sepia pseudoacacia includes such steps as temporarily culturing the sepia pseudoacacia in an indoor culture pool with egg-attached base, stimulating by flowing water, feeding the bait containing fermented soybean dregs, collecting fertilized eggs, artificial incubation, culturing the incubated seedlings in seedling pool, culturing in culture pool, and fermenting the fermented soybean dregs with rhamnose lactobacillus containing tocopherol and cysteine.
The egg-attached base is a polyethylene mesh sheet with a mesh of 27 mm.
The preparation method of the adlay base comprises the following steps: fully mixing polyethylene and epsilon-polylysine with the weight ratio of 100:1.3 on a double-roll mixing mill at 160 ℃ and 300rpm for 10min, then putting the mixed composite material into a die, transferring the die onto a flat vulcanizing machine, carrying out hot pressing at 160 ℃ and 10MPa for 5min, then carrying out cold pressing at room temperature, and taking out the die after the cold pressing is finished, thus obtaining the polyethylene mesh sheet attached egg base. The epsilon-polylysine contains 0.04 wt% of D-lysine.
The specific fermentation steps of the fermented soybean meal are as follows: grinding soybean meal to 100 meshes, adding water until the water content is 50%, uniformly mixing, autoclaving for 25min, cooling, inoculating lactobacillus rhamnosus with an inoculum size of 4%, fermenting at 38 deg.C for 1.5d, freeze drying, pulverizing, and sieving with 250 mesh sieve to obtain fermented soybean meal.
The tocopherol accounts for 4.0 percent of the weight of the lactobacillus rhamnosus, and the cysteine accounts for 1.1 percent of the weight of the lactobacillus rhamnosus.
The bait comprises the following components in parts by weight: 22 parts of fermented soybean meal, 15 parts of fish meal, 15 parts of bone meal, 10 parts of mysidacea, 0.003 part of vitamin A, 0.0015 part of vitamin E, 0.003 part of vitamin C, 3-7 parts of vegetable oil, 0.3 part of citric acid and 0.2 part of phytase.
The conditions for parent breeding were: the water temperature is 22 deg.C, the salinity of seawater is 28, the photoperiod L: D is 4:20, the flow velocity of flowing water stimulation is 6m/min, the time is 12min, the bait is thrown for 3 times a day, and the total feeding amount is 0.45 times of the parent weight.
The parent selection and matching specifically comprises the following steps: according to the natural breeding method of the sepia pseudosciaenae, healthy and sexually mature male and female sepia pseudosciaenae are respectively selected as parents, and then the male and female sepia pseudosciaenae are matched according to the ratio of 3: 1.
The artificial incubation comprises the following specific steps: and (3) placing the fertilized eggs in a culture pond, controlling the water temperature of the culture pond to be 22 ℃, the salinity of seawater to be 28, the water-soluble oxygen content to be more than or equal to 6mg/L and the incubation time to be 30d, and covering a single-layer sunshade net above the incubation pond.
Example 3:
a method for improving the fertility of sepia pseudoacacia includes such steps as temporarily culturing the sepia pseudoacacia in an indoor culture pool with egg-attached base, stimulating by flowing water, feeding the bait containing fermented soybean dregs, collecting fertilized eggs, artificial incubation, culturing the incubated seedlings in seedling pool, culturing in culture pool, and fermenting the fermented soybean dregs with rhamnose lactobacillus containing tocopherol and cysteine.
The egg-attached base is a polyethylene mesh sheet with a mesh of 25 mm.
The preparation method of the adlay base comprises the following steps: and (2) fully mixing polyethylene and epsilon-polylysine in a weight ratio of 100:3 on a double-roll mixing mill at 158 ℃ and 350rpm for 8min, then putting the mixed composite material into a mold, transferring the mold to a flat vulcanizing machine, carrying out hot pressing at 163 ℃ and 8MPa for 6min, then carrying out cold pressing at room temperature, and taking out the mold after the cold pressing is finished, thus obtaining the polyethylene mesh sheet attached egg base. The epsilon-polylysine contains 0.03 wt% of D-lysine.
The specific fermentation steps of the fermented soybean meal are as follows: grinding soybean meal to 120 meshes, adding water until the water content is 45%, uniformly mixing, autoclaving for 30min, cooling, inoculating lactobacillus rhamnosus with the inoculation amount of 3%, fermenting at 40 deg.C for 1d, freeze drying, pulverizing, and sieving with 300 meshes sieve to obtain fermented soybean meal.
The tocopherol accounts for 3.5 percent of the weight of the lactobacillus rhamnosus, and the cysteine accounts for 1.5 percent of the weight of the lactobacillus rhamnosus.
The bait comprises the following components in parts by weight: 18 parts of fermented soybean meal, 20 parts of fish meal, 10 parts of bone meal, 15 parts of mysidacea powder, 0.002 part of vitamin A, 0.002 part of vitamin E, 0.002 part of vitamin C, 7 parts of vegetable oil, 0.2 part of citric acid and 0.3 part of phytase.
The conditions for parent breeding were: the water temperature is 20 deg.C, the salinity of seawater is 30, the photoperiod L: D is 4:18, the flow velocity of flowing water stimulation is 8m/min, the time is 10min, the bait is thrown for 4 times a day, and the total feeding amount is 0.3 times of the parent weight.
The parent selection and matching specifically comprises the following steps: according to the natural breeding method of the sepia pseudosciaenae, healthy and sexually mature male and female sepia pseudosciaenae are respectively selected as parents, and then the male and female sepia pseudosciaenae are matched according to the ratio of 4: 1.
The artificial incubation comprises the following specific steps: and (3) placing the fertilized eggs in a culture pond, controlling the water temperature of the culture pond to be 25 ℃, the salinity of seawater to be 25, the water-soluble oxygen content to be more than or equal to 6mg/L and the incubation time to be 35d, and covering a single-layer sunshade net above the incubation pond.
Example 4:
the method for improving the reproductive capacity of the sepia lycidas comprises the following steps: epsilon-polylysine contained no D-lysine, and the others were identical to those of example 2.
Example 5:
the method for improving the fertility of the sepia lycidas is characterized in that fermented soybean meal is fermented by lactobacillus rhamnosus which does not contain tocopherol and cysteine, and the rest is completely consistent with the embodiment 2.
Example 6:
the average amount of egg-attached (pellets), hatching rate (%), and survival rate (%) were calculated by setting example 2 as the test group and example 4 and example 5 as the control group 1 and control group 2, respectively. The calculation results are shown in fig. 1.
As can be seen from fig. 1, the amount of egg attachment, the hatching rate and the survival rate of the test group are all higher than those of the control group 1 and the control group 2, which indicates that the reproductive capacity of the sepia pseudobulb is higher than that of the example 4 and the example 5, and further indicates that the addition of epsilon-polylysine can improve the amount of egg attachment of the sepia pseudobulb, and further improve the hatching rate; the special existence of the tocopherol and the cysteine can improve the hatchability and the survival rate of the sepia pseudobulb and finally improve the reproductive capacity of the sepia pseudobulb.
Conventional operations in the operation steps of the present invention are well known to those skilled in the art and will not be described herein.
The embodiments described above are intended to illustrate the technical solutions of the present invention in detail, and it should be understood that the above-mentioned embodiments are only specific embodiments of the present invention, and are not intended to limit the present invention, and any modification, supplement or similar substitution made within the scope of the principles of the present invention should be included in the protection scope of the present invention.
Claims (5)
1. The method for improving the fertility of the sepia lycidas is characterized by comprising the following steps of: temporarily culturing the combined sepia mimosoides in an indoor culture pond provided with an egg-attached base, carrying out flowing water stimulation, feeding bait containing fermented soybean meal, collecting fertilized eggs for artificial incubation after the parents of the sepia mimosoides naturally lay eggs, then culturing the incubated seedlings in a seedling pond, culturing the seedlings in the culture pond after the trunk of the seedlings grows to be qualified, and fermenting the fermented soybean meal by using lactobacillus rhamnosus containing tocopherol and cysteine;
the egg-attached base is a polyethylene mesh sheet with a mesh of 25-30 mm;
fully mixing polyethylene and epsilon-polylysine with the weight ratio of 100:0.1-3 on a double-rod mixing mill at 163 ℃ under 158-;
the epsilon-polylysine contains 0.03 to 0.06 weight percent of D-lysine;
the specific fermentation steps of the fermented soybean meal are as follows: grinding the soybean meal to 80-120 meshes, adding water until the water content is 45-53%, uniformly mixing, then sterilizing at high pressure for 20-30min, cooling, inoculating lactobacillus rhamnosus with the inoculation amount of 3-5%, fermenting for 1-2d at 35-40 ℃, freeze drying, crushing, and sieving with a 200-mesh and 300-mesh sieve to obtain fermented soybean meal;
the tocopherol accounts for 3.5-5.6% of the weight of the lactobacillus rhamnosus, and the cysteine accounts for 0.8-1.5% of the weight of the lactobacillus rhamnosus.
2. The method for improving the fertility of sepia pseudobulb according to claim 1, wherein: the bait comprises the following components in parts by weight: 18-28 parts of fermented soybean meal, 10-20 parts of fish meal, 10-20 parts of bone meal, 7-15 parts of shrimp bran powder, 0.002-0.004 part of vitamin A, 0.001-0.002 part of vitamin E, 0.002-0.005 part of vitamin C, 3-7 parts of vegetable oil, 0.2-0.4 part of citric acid and 0.1-0.3 part of phytase.
3. The method for improving the fertility of sepia pseudobulb according to claim 1, wherein: the conditions for parent culture are as follows: the water temperature is 20-25 deg.C, the salinity of seawater is 25-30, the photoperiod L: D is 4:18-25, the flow velocity of flowing water stimulation is 5-8m/min, the time is 10-15min, the bait is thrown for 2-4 times daily, and the total feeding amount is 0.3-0.6 times of the parent weight.
4. The method for improving the fertility of sepia pseudobulb according to claim 1, wherein: the parent selection and matching specifically comprises the following steps: according to the natural breeding method of the sepia pseudosciaenae, healthy and sexually mature male and female sepia pseudosciaenae are respectively selected as parents, and then the male and female sepia pseudosciaenae are matched according to the ratio of 2-4: 1.
5. The method for improving the fertility of sepia pseudobulb according to claim 1, wherein: the artificial incubation specifically comprises the following steps: putting the fertilized eggs into a culture pond, controlling the water temperature of the culture pond to be 20-25 ℃, the salinity of seawater to be 25-30, the water-soluble oxygen content to be more than or equal to 6mg/L, and the incubation time to be 20-35d, and covering a single-layer sunshade net above the incubation pond.
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| JPH10136828A (en) * | 1996-11-13 | 1998-05-26 | Asahi Chem Ind Co Ltd | Fish bank and substrate therefor |
| CN102823529A (en) * | 2012-09-17 | 2012-12-19 | 湛江师范学院 | Sepia lycidas fry breeding method |
| CN102939924A (en) * | 2012-11-13 | 2013-02-27 | 宁波大学 | Method for artificially breeding sepia lycidas gray |
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