CN115886159B - Gonad development strengthening bait and hybridization method for puffer fish and puffer fish hexaflumuron - Google Patents
Gonad development strengthening bait and hybridization method for puffer fish and puffer fish hexaflumuron Download PDFInfo
- Publication number
- CN115886159B CN115886159B CN202310218875.8A CN202310218875A CN115886159B CN 115886159 B CN115886159 B CN 115886159B CN 202310218875 A CN202310218875 A CN 202310218875A CN 115886159 B CN115886159 B CN 115886159B
- Authority
- CN
- China
- Prior art keywords
- fish
- injection
- female
- puffer fish
- male
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Fodder In General (AREA)
- Feed For Specific Animals (AREA)
Abstract
The invention provides a parent fish gonad development strengthening bait and a hybridization method of male puffer fish and female puffer fish, belonging to the technical field of aquaculture. The parent fish gonad development strengthening bait comprises the following components in percentage by mass: 65% of fish feed, 20% of fresh shrimp meat, 5% of propagation enhancing additive, 5% of fish oil powder, 2.5% of vitamin C, 1.8% of vitamin E and 0.7% of taurine. The strengthening bait can promote gonad development and sexual maturity of parent fish and improve the quality of parent fish sperms and ova; according to the invention, the male puffer fish and female puffer fish are combined with sperm and eggs for the first time in China by a special artificial fertilization mode, so that high-quality hybrid fertilized eggs are obtained, and a certain number of seedlings are obtained through later-stage seedling cultivation. The realization of the technical scheme of the invention promotes the diversified development of the cultivated varieties in China, and has important social significance for improving the fishery industry structure and promoting the industrial economic development.
Description
Technical Field
The invention belongs to the technical field of aquaculture, and particularly relates to a parent fish gonad development strengthening bait and a hybridization method of male puffer fish and female puffer fish.
Background
Both the takifugu obscurus (Diodon holocanthus) and the takifugu hexapetri (Diodon holocanthus) are of the class of the finfish, the order of the tetrodotodes, one of the families of the Diodonidae, commonly known as the angry young, the thorn gauge and the like, and the tropical marine bottom fishes mainly inhabit the shallow sea reef area, the soft bottom sea area or the open water area. The fish skin is mainly used for predating invertebrates such as mollusks, sea urchins, living crabs and crabs at night, the meat is delicious, the fish skin is rich in rich collagen, and the fish skin is popular in the market. The puffer fish has large individual and average weight of about 3000g, but the wild parent fish has unstable quantity and poor disease resistance, and is difficult to popularize on a large scale. Although the individual of the fugu hexapustus is small, the average weight is about 500g, but the disease resistance is strong, and the population quantity is large.
The hybridization of the puffer fish and the puffer fish can further promote the diversified development of the cultivated varieties in China, and has important social significance for improving the industrial structure of fishery and promoting the economic development of industry. At present, the bait for promoting the gonad development of parent fish is mainly effective only for one organism, so that the hybridization of the puffer fish and the puffer fish is realized, and one of the key problems comprises how to ensure that the gonad development of the parent fish of the puffer fish and the puffer fish is good at the same time, and continuously and stably supply high-quality fertilized eggs.
Disclosure of Invention
The invention aims to provide a parent fish gonad development strengthening bait for promoting gonad development of takifugu obscurus and takifugu hexaflumus, which can promote gonad development of takifugu obscurus and takifugu hexaflumus simultaneously.
The invention provides parent fish gonad development strengthening bait which comprises the following components in percentage by mass:
65% of fish feed, 20% of fresh shrimp meat, 5% of propagation enhancing additive, 5% of fish oil powder, 2.5% of vitamin C, 1.8% of vitamin E and 0.7% of taurine.
Preferably, the propagation enhancing additive comprises the following raw materials: cuttlefish viscera powder, shell-kettle algae powder, marine alcohol mother, compound vitamin, vitamin E acetate, soybean lecithin and antarctic krill powder.
The invention provides a hybridization method of male puffer fish and female puffer fish, which comprises the following steps:
culturing the male puffer fish and the female puffer fish with the enhanced bait according to the technical scheme;
ripening the male and female puffer fish subjected to nutrition-enhanced culture to obtain sexually mature male and female puffer fish;
manually collecting sperms of male puffer fish and ova of female puffer fish, and combining sperm and ova to obtain fertilized ova;
incubating the fertilized eggs.
Preferably, the ripening method of female globefish is to inject chorionic gonadotrophin and luteinizing hormone releasing hormone A2;
the injection includes a first injection and a second injection;
the injection quantity of chorionic gonadotrophin in the first injection is 230-245 units/kg; the injection quantity of the luteinizing hormone releasing hormone A2 is 2.5-2.8 mug/kg;
the injection amount of chorionic gonadotrophin in the second injection is 350-360 units/kg; the injection quantity of the luteinizing hormone releasing hormone A2 is 4.4-4.5 mug/kg.
Preferably, the ripening method of the male puffer fish is to inject luteinizing hormone releasing hormone A2;
the injection includes a first injection and a second injection;
the injection quantity of the luteinizing hormone releasing hormone A2 in the first injection is 2-2.2 mu g/kg;
and the injection quantity of the luteinizing hormone releasing hormone A2 in the second injection is 4-4.3 mu g/kg.
Preferably, the first injection and the second injection are separated by a time of 24 hours.
Preferably, when the sperms and the ovum are mixed, the mass ratio of the sperms of the male puffer fish to the ovum of the female puffer fish is 0.5mL:500g.
Preferably, the water temperature of the nutrition-enriched culture is 26-28 ℃; the salinity of the nutrition-enriched culture is 28-30; the time of the nutrient enrichment culture is 45-50 d.
Preferably, the temperature of the seawater for hatching fertilized eggs is 25-26 ℃, and the salinity of the seawater is 28-30.
The invention has the beneficial effects that:
the invention provides parent fish gonad development strengthening bait which comprises the following components in percentage by mass: 65% of fish feed, 20% of fresh shrimp meat, 5% of propagation enhancing additive, 5% of fish oil powder, 2.5% of vitamin C, 1.8% of vitamin E and 0.7% of taurine. The parent fish gonad development strengthening bait provided by the invention can promote the gonad development of female puffer fish and male puffer fish parent fish, promote the sexual maturity of parent fish and improve the quality of parent fish sperms and ova; according to the invention, the male puffer fish and female puffer fish are combined with sperm and eggs for the first time in China in a special artificial fertilization mode, so that high-quality hybrid fertilized eggs are obtained, and a certain number of offspring seeds are obtained through later-stage offspring seed cultivation. The realization of the hybridization technology of the puffer fish and the puffer fish promotes the diversified development of the cultivated varieties in China, and has important social significance for improving the industrial structure of fishery and promoting the economic development of industry.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a picture of a male puffer fish selected in example 2;
FIG. 2 is a picture of female puffer fish selected in example 2;
FIG. 3 is a view of a fertilized egg obtained in example 2 under a microscope;
FIG. 4 is a diagram of an initially hatched fish in example 2;
fig. 5 is an external packing diagram of the fish feed;
FIG. 6 is a pellet view of fish feed.
Detailed Description
The invention provides parent fish gonad development strengthening bait which comprises the following components in percentage by mass:
65% of fish feed, 20% of fresh shrimp meat, 5% of propagation enhancing additive, 5% of fish oil powder, 2.5% of vitamin C, 1.8% of vitamin E and 0.7% of taurine.
The present invention is not limited to the sources of the components, and may be practiced using commercially available products known to those skilled in the art, unless otherwise specified.
In the present invention, the feed for fish is preferably purchased from the company of both forestry and plant, the outer package of the purchased feed for fish is shown in fig. 5, and the specific pellet of the feed for fish is shown in fig. 6.
In the present invention, the propagation enhancing additive preferably includes squid viscera powder, column-kettle seaweed powder, marine alcohol master, compound vitamin, vitamin E acetate, soybean lecithin, and euphausia superba powder. In the invention, the propagation enhancing additive comprises, by mass, 30% of cuttlefish viscera powder, 20% of column-kettle algae powder, 20% of marine alcohol master batch, 5% of compound vitamin, 5% of vitamin E acetate, 10% of soybean lecithin and 10% of antarctic krill powder. In the present invention, the propagation enhancing additive is preferably purchased from the company of both woods and plants.
The parent fish gonad development strengthening bait provided by the invention is prepared on site. In the invention, the preparation method of the parent fish gonad development enhancement bait is preferably to granulate the raw materials after mixing the raw materials. In the present invention, the particle size of the granulated particles is preferably 0.5 to 0.6cm, more preferably 0.5cm. The method of granulating the present invention is not particularly limited, and conventional granulating methods in the art may be employed. The invention prepares the parent fish gonad development strengthening bait into pellet feed to ensure that the nutrition components cannot be lost or changed.
The parent fish gonad development strengthening bait provided by the invention can promote the gonad development of male puffer fish and female puffer fish, promote the sexual maturation of parent fish and improve the quality of parent fish sperms and ova, thereby continuously and stably supplying high-quality fertilized ova, realizing the hybridization of puffer fish and puffer fish, and obviously improving the artificial fertilization rate.
In the invention, the use method of the parent fish gonad development enhancement bait preferably comprises the following steps: feeding parent fish gonad development strengthening baits in a feeding way, wherein the feeding times are 2 times per day; the 2 feeds are preferably performed at 8:00 a.m. and 12:00 a.m..
The invention provides a hybridization method of male puffer fish and female puffer fish, which comprises the following steps:
culturing the male puffer fish and the female puffer fish with the enhanced bait according to the technical scheme;
ripening the male and female puffer fish subjected to nutrition-enhanced culture to obtain sexually mature male and female puffer fish;
manually collecting sperms of male puffer fish and ova of female puffer fish, and combining sperm and ova to obtain fertilized ova;
incubating the fertilized eggs.
The invention adopts the reinforced bait for carrying out nutrition reinforced culture on male puffer fish and female puffer fish.
In the invention, the male puffer fish and the female puffer fish are preferably selected parent fish; the selection criteria are preferably body surface intact and swimming normal. In the invention, the weight of the male puffer fish is preferably more than or equal to 3kg; the weight of the female puffer fish is preferably more than or equal to 1kg. In the present invention, the male and female puffer fish are preferably purchased from wild puffer fish and wild puffer fish captured in the sea area of south sea. In the invention, the parent fish is fed by using running water during transportation. When the invention is used for nutrition-enriched culture, the culture water of the parent fish is preferably seawater filtered by a dark sediment and sand filter. In the invention, the water temperature of the nutrient enrichment culture is preferably 26-28 ℃; the salinity of the nutrient enrichment culture is preferably 28-30. In the invention, the water temperature and the salinity are favorable for promoting the gonad development of the fugu and facilitating the oviposition of the fugu when the nutrition-enhanced culture is carried out. The method for nutrition-enhanced culture of the invention is to feed the parent fish gonad development-enhanced bait in a feeding way. In the invention, the parent fish gonad development strengthening bait is preferably fed for 2 times per day; the 2 feeds are preferably performed at 8:00 a.m. and 12:00 a.m.. The two parent fish nutrition enrichment cultures are carried out simultaneously, and the time of the nutrition enrichment culture is preferably 45-50 d, more preferably 50d. In the present invention, the nutrient enrichment culture is preferably performed in a culture pond. The invention promotes the sexual maturity of parent fish through early-stage nutrition-enriched culture, and improves the quality of parent fish sperms and ova.
After the nutrition-enhanced culture of the male and female puffer fish, the invention ripening the male and female puffer fish to obtain sexually ripe male and female puffer fish.
The method for ripening female puffer fish is preferably injection of gonadotrophin. In the present invention, the gonadotrophin preferably includes chorionic gonadotrophin and luteinizing hormone releasing hormone A2. The number of injections according to the present invention is preferably 2, the first injection and the second injection, respectively. In the invention, the injection amount of chorionic gonadotrophin in the first injection is preferably 230-245 units/kg, more preferably 235 units/kg; the injection quantity of the luteinizing hormone releasing hormone A2 is preferably 2.5-2.8 mug/kg, and more preferably 2.6 mug/kg. In the invention, the injection amount of chorionic gonadotrophin in the second injection is preferably 350-360 units/kg, more preferably 350 units/kg; the injection quantity of the luteinizing hormone releasing hormone A2 is preferably 4.4-4.5 mug/kg, and more preferably 4.5 mug/kg. In the present invention, the interval time between the first injection and the second injection is preferably 24 hours.
The method for ripening male puffer fish is preferably injection of gonadotrophin. In the present invention, the gonadotropin-developing hormone preferably comprises luteinizing hormone releasing hormone A2. The number of injections according to the present invention is preferably 2, the first injection and the second injection, respectively. In the invention, the injection amount of the luteinizing hormone releasing hormone A2 in the first injection is preferably 2-2.2 mug/kg, and more preferably 2.2 mug/kg. In the invention, the injection amount of the luteinizing hormone-releasing hormone A2 in the second injection is preferably 4-4.3 mug/kg, and more preferably 4.3 mug/kg. In the present invention, the interval time between the first injection and the second injection is preferably 24 hours.
The sources of chorionic gonadotrophin and luteinizing hormone releasing hormone A2 in gonadotrophin are not particularly limited in the invention, and the products are generally commercially available in the art. In the present invention, chorionic gonadotrophin and luteinizing hormone releasing hormone A2 among said gonadotrophin are purchased from Ningbo Sansheng biotechnology company.
The ripening of male and female puffer fish is preferably carried out synchronously. The injection according to the invention is preferably a tail intramuscular injection. The injection time is preferably 8:00-9:00. The ripening method adopted by the invention can improve the synchronous rate of gonadal maturation of parent fish and improve the fertilization rate.
After the second injection of the female globefish, the invention preferably observes the female globefish every 2 hours. The invention observes that the outer edge of the female fecund hole of the puffer fish is reddish, when the female fecund hole of the female puffer fish is loosened, a straw is gently inserted into the female fecund hole of the female puffer fish, and a few ova are taken for microscopic examination. The invention detects whether female puffer fish reaches sexual maturity or not through microscopic examination.
After sexually mature male and female puffer fish are obtained, the invention artificially collects sperms of male puffer fish and ova of female puffer fish, and carries out sperm-egg combination to obtain fertilized eggs.
The invention preferably uses scissors to cut off thorns on the abdomen of the parent fish before the parent fish is subjected to sperm and ovum extraction. The invention cuts off the thorns on the abdomen of the parent fish, which mainly prevents the thorns from causing inconvenience for later-stage sperm and egg taking. After the sexually mature male and female puffer fish are obtained, the invention artificially collects the sperms of the male puffer fish and the ovum of the female puffer fish. The parent fish is preferably subjected to micro anesthesia before the parent fish sperm and ovum are collected. The micro anesthesia of the invention preferably adopts eugenol water solution; the volume ratio of eugenol to water in the eugenol water solution is preferably 1mL:1L. In the present invention, the water used in the aqueous eugenol solution is preferably seawater for breeding parent fish. The method for carrying out micro anesthesia on the parent fish is preferably to place the parent fish in a eugenol water solution for 2-3 min, more preferably for 2min; preferably, the aqueous eugenol solution is subjected to micro-aeration during the said placing.
After the parent fish is micro-anesthetized, the invention preferably extracts the male puffer fish and simultaneously extracts the female puffer fish.
The method of the present invention is not particularly limited, and methods known in the art may be used. The method for extracting the sperms of the male puffer fish is preferably to put the sides of the male puffer fish in an anatomical plate, and cover the head with a wet towel; wiping the genital orifice with dry paper towel, and slightly extruding with thumb and index finger at the positions of about 7cm above the genital orifice and facing the genital orifice; and when the white liquid flows out, sucking the white liquid by using a suction pipe to obtain the sperms. The obtained sperms are stored in a centrifuge tube; the preservation temperature is preferably 6-8 ℃. In the invention, the parent fish after the semen collection is preferably cultured independently. The method for taking the eggs of female puffer fish is preferably artificial extrusion ovulation. The artificial extrusion ovulation method is that female puffer fish is placed in an anatomical disc at the side, and the female puffer fish is wrapped towards the head 10cm above a reproduction hole by a wet towel; wiping the genital orifice with a dry paper towel, and then slightly and slowly extruding from the 10cm position of the genital orifice towards the direction of the genital orifice; when the ovum flows out, the ovum collecting utensil is placed under the reproduction hole to collect the ovum. The parent fish after ovum taking is preferably cultured in a culture pond separately. The invention carries out unified cultivation on male puffer fish and female puffer fish in the nutrition-enhanced cultivation stage, and carries out independent cultivation on parent fish after sperm and egg taking, thereby being convenient for distinguishing and observing the later-stage state.
After the collected sperms and ova are obtained, the invention performs sperms and ova combination. In the invention, the sperm-egg combination is preferably carried out by mixing the sperm and the ovum obtained in the technical scheme; the mixing is preferably performed by pouring the extracted sperm into a vessel containing the extracted egg. When the sperm and the ovum are mixed, the mass ratio of the sperm volume of the male puffer fish to the ovum of the female puffer fish is preferably 0.5mL:500g. When sperm-egg combination is carried out, stirring is preferably carried out; the stirring mode is preferably gentle stirring by using feathers; the stirring time is preferably 1min. In the present invention, the stirring is advantageous for the complete binding of sperm and eggs. After stirring, the invention preferably slowly adds water to the fertilized egg; the amount of water added is preferably 1/3 of the fertilized egg volume. In the invention, the water temperature of the water is preferably 25-26 ℃; the salinity of the water is preferably 28-30. In the present invention, the water is preferably seawater filtered through a dark sediment, sand filter. After adding 1/3 of the fertilized egg volume of water, the present invention preferably continues the above stirring. After the completion of the stirring, sea water containing fertilized eggs was obtained. The mode of semen collection, ovum collection and fertilized ovum of the invention obviously improves the artificial fertilization efficiency, simultaneously improves the fertilization rate, and solves the problem of fertilization limitation different from other fishes due to thorns on the body surface of the puffer fish.
After fertilized eggs are obtained, the fertilized eggs are hatched.
After fertilized eggs are obtained, the invention pours seawater containing fertilized eggs into a hatching barrel for hatching. The water temperature for hatching is preferably 25-26 ℃; the salinity of hatching is preferably 28-30.
In the invention, the devices used in the hybridization process are stored in an incubator with the temperature of 6-8 ℃ after being sterilized.
The invention promotes the sexual maturity of the parent by the early-stage nutrition-enriched culture and improves the quality of the parent sperms and ova. According to the invention, the gonadal hormone is injected for several times, so that the synchronous rate of parent gonadal maturation can be improved, and the fertilization rate can be improved; compared with other fishes, the artificial insemination is different because of the thorns on the body surface, and the artificial insemination efficiency and the insemination rate are obviously improved by adopting the methods of semen collection, ovum collection and fertilized ovum.
Test results show that 3 batches of fertilized eggs artificially hybridized with the male and female puffer fish are obtained by the hybridization method of the male and female puffer fish provided by the invention, the total quantity of the fertilized eggs is about 40.8 ten thousand grains, the fertilized eggs (shown in figure 3) are obtained by about 25.4 ten thousand grains, and the fertilization rate is 62.2%. The number of the initially hatched fries is about 16.5 ten thousand tails (as shown in figure 4), the average body length is about 3mm, and the hatching rate reaches 65 percent.
The technical solutions provided by the present invention are described in detail below with reference to the drawings and examples for further illustrating the present invention, but they should not be construed as limiting the scope of the present invention.
The devices used in the following technical proposal are all stored in an incubator with the temperature of 6 ℃ to 8 ℃ after being disinfected.
The seawater used in the following technical proposal is the seawater filtered by the dark sediment and sand filter.
Example 1
The parent fish gonad development strengthening bait comprises the following components in percentage by mass:
65% of fish feed, 20% of fresh shrimp meat, 5% of propagation enhancing additive, 5% of fish oil powder, 2.5% of vitamin C, 1.8% of vitamin E and 0.7% of taurine.
The feed for the fish is purchased from a forest and plant company, the outer package of the purchased feed for the fish is shown in figure 5, and the specific pellet diagram of the feed for the fish is shown in figure 6; propagation enhancing additives were purchased from woodland co.
The propagation strengthening additive comprises the following components: cuttlefish viscera powder, shell-kettle algae powder, marine alcohol mother, compound vitamin, vitamin E acetate, soybean lecithin and antarctic krill powder;
the contents of the components in mass percent are shown in table 1.
TABLE 1 ratio of ingredients of propagation-enhancing additives
Example 2
A hybridization method of male puffer fish and female puffer fish comprises the following steps:
parent fish selection
Parent fish is derived from wild puffer fish and puffer fish captured in the sea area of south China sea purchased from the wharf of the Tan gate of Hainan. Culturing male puffer fish (shown in figure 1) with body surface intact, normal swimming and body weight above 3000g and female puffer fish (shown in figure 2) with body surface intact, normal swimming and body weight above 1000 g.
Nutrient enrichment culture
When nutrition-enriched culture is carried out, the seawater filtered by the dark sediment and the sand filter is taken as the culture water for parent fish; the water temperature of the seawater is 26-28 ℃, and the salinity is 28-30. The specific method for nutrient enrichment culture comprises the following steps: the parent fish gonad development enhancing bait in example 1 was used to feed male and female puffer fish for satiation twice daily at 8:00 a.m. and 12:00 a.m. The time of nutrient enrichment culture is 50d.
Ripening acceleration
In order to ensure synchronous maturation of parent fish ovum and sperm, the parent fish is ripened by the method specifically comprises the following steps:
the synchronous tail muscle injection gonadotrophin of male and female puffer fish comprises the following steps:
the ripening method of female globefish is to inject chorionic gonadotrophin and luteinizing hormone releasing hormone A2, and the injection times are 2 times. In the first injection, the injection is carried out according to 235 units/kg of chorionic gonadotrophin, 2.6 mug/kg of luteinizing hormone releasing hormone A, and the injection time is 8:00-9:00. At 24h intervals, a second injection was performed in which injections of chorionic gonadotrophin at 350 units/kg and luteinizing hormone releasing hormone A2 at 4.5 μg/kg were performed.
The ripening method of the male puffer fish is to inject luteinizing hormone releasing hormone A2, and the injection times are 2 times. In the first injection, the lutein-releasing hormone A2 is injected at the rate of 2.2 mug/kg, and the injection time is 8:00-9:00. At 24h intervals, a second injection was performed in which injections of luteinizing hormone-releasing hormone A2 were made at 4.3 μg/kg.
Selecting sexually mature parent fish
Female globefish individuals were examined every 2 hours after the second gonad hormone injection. When the outer edge of the female genital hole of the puffer fish is observed to be reddish, when the female genital hole is loosened, a straw is gently inserted into the female genital hole to take a few ova, and whether sexual maturity is reached or not is checked by a microscope. If mature, the artificial extrusion ovulation can be performed.
After sexually mature male and female puffer fish, sperm and ovum were collected.
Before collection of sperm and ovum, the male and female puffer fish are subjected to micro anesthesia. The method for micro anesthesia specifically comprises the following steps:
placing parent fish in eugenol seawater solution for 2min, and micro-aerating in eugenol seawater solution. The preparation method of the eugenol seawater solution comprises the following steps: eugenol and seawater are mixed according to the volume ratio of 1mL:1L of the components were mixed together.
After the parent fish is micro-tingled, the male and female puffer fish are respectively subjected to sperm and egg taking, and before the sperm and egg taking, the abdominal thorns are subtracted by scissors.
The semen extraction method comprises the following steps:
placing the side of the male puffer fish in an anatomical plate, and covering the head with a wet towel; the genital opening was wiped dry with a dry paper towel and gently pressed toward the genital opening with thumb and index finger at about 7cm sides above the genital opening. When the white liquid flows out, the white liquid is timely sucked by a suction pipe, and the semen is stored in an incubator with the temperature of 6-8 ℃ for standby. And (5) putting the parent fish after the semen collection back into a culture pond for independent culture.
The method for taking the ovum comprises the following steps:
the female puffer hexa is placed laterally in an anatomical tray and wrapped with a wet towel 10cm above the reproduction aperture toward the head. The genital orifice was wiped dry with a dry paper towel and then gently pressed from 10cm toward the orifice. When the ovum flows out, the ovum collecting vessel is placed under the reproduction hole for collection. And (5) putting the parent fish after ovum taking back into a culture pond for independent culture.
After sperm and ovum are obtained, sperm-ovum combination is performed. The sperm-egg combination method comprises the following steps:
pouring the sperms which are picked in advance into a utensil filled with eggs, wherein the mass ratio of the volume of the sperms to the mass of the eggs is 0.5mL:500g. Mixing sperm and egg, and lightly stirring with feather for 1min to fully combine sperm and egg; slowly adding 1/3 of the fertilized egg volume of seawater, wherein the water temperature of the seawater is 25-26 ℃, the salinity of the seawater is 28-30, and continuously lightly stirring the seawater for 1min by using feathers to obtain the seawater containing fertilized eggs.
And pouring the seawater containing fertilized eggs into a hatching barrel for hatching, wherein the temperature of the hatching water is 25-26 ℃, and the salinity is 28-30.
Hybridization of male and female Takifugu obscurus was performed as described in example 2 to obtain 3 batches of fertilized eggs of artificial hybridization of male and female Takifugu obscurus, the first batch being 2022, 5 months and 6 days, 3 tails of male Takifugu obscurus and 6 tails of female Takifug obscurus; the second batch time is 2022, 5 months and 10 days, 4 tails of male puffer fish and 7 tails of female puffer fish; the third batch time is 2022, 6 months and 12 days, 3 tails of male puffer fish and 5 tails of female puffer fish; the total fertilized egg amount is 40.8 ten thousand grains, the fertilized eggs (as shown in figure 3 and a 40-fold enlarged observation chart of a microscope) are obtained, and the fertilized rate is 62.2%. The length of the initially hatched fries (shown in figure 4) is about 3mm on average, the number of the initially hatched fries is about 16.5 ten thousand, and the hatching rate reaches 65%.
Culturing the initially hatched fries for 60 days, wherein the average growth speed of the 60-day-old hybrid puffer fish fries is 0.9mm/d; the average growth rate of the seeds is improved by more than 14.3 percent compared with that of the seeds of the fugu hexaflumuron.
In conclusion, the parent fish gonad development strengthening bait provided by the invention can promote parent fish to mature, and improve the quality of parent fish sperms and ova. The invention can improve the synchronous rate of parent gonad maturation and the fertilization rate through nutrition-enhanced culture and the hybridization method of male puffer fish and female puffer fish. Meanwhile, the growth rate of the hybrid puffer fish obtained by the hybridization method provided by the invention is obviously improved relative to that of the puffer fish hexaflumuron.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (5)
1. A method for hybridizing male puffer fish with female puffer fish, comprising the steps of:
culturing male and female puffer fish with gonad development strengthening bait for parent fish;
the parent fish gonad development strengthening bait comprises the following components in percentage by mass:
65% of fish feed, 20% of fresh shrimp meat, 5% of propagation strengthening additive, 5% of fish oil powder, 2.5% of vitamin C, 1.8% of vitamin E and 0.7% of taurine;
the propagation enhancing additive comprises the following raw materials: cuttlefish viscera powder, shell-kettle algae powder, marine alcohol mother, compound vitamin, vitamin E acetate, soybean lecithin and antarctic krill powder;
the propagation strengthening additive comprises, by mass, 30% of cuttlefish viscera powder, 20% of column-kettle algae powder, 20% of marine alcohol master batch, 5% of compound vitamin, 5% of vitamin E acetate, 10% of soybean lecithin and 10% of antarctic krill powder;
ripening the male and female takifugu obscurus after nutrition-enhanced cultivation to obtain sexually ripe male and female takifugu obscurus;
the method for ripening female globefish comprises injecting chorionic gonadotrophin and luteinizing hormone releasing hormone A2;
the injection includes a first injection and a second injection;
the injection quantity of chorionic gonadotrophin in the first injection is 230-245 units/kg; the injection quantity of the luteinizing hormone releasing hormone A2 is 2.5-2.8 mug/kg;
the injection quantity of chorionic gonadotrophin in the second injection is 350-360 units/kg; the injection quantity of the luteinizing hormone releasing hormone A2 is 4.4-4.5 mug/kg;
the ripening method of the male puffer fish is to inject luteinizing hormone releasing hormone A2;
the injection includes a first injection and a second injection;
the injection quantity of the luteinizing hormone releasing hormone A2 in the first injection is 2-2.2 mug/kg;
the injection quantity of the luteinizing hormone releasing hormone A2 in the second injection is 4-4.3 mug/kg
Manually collecting sperms of male puffer fish and ova of female puffer fish, and combining sperm and ova to obtain fertilized ova;
incubating the fertilized eggs.
2. The hybridization method according to claim 1, wherein the first injection and the second injection are separated by a time of 24 hours.
3. The hybridization method according to claim 1, wherein the mass ratio of the sperm volume of the male puffer fish to the ovum mass of the female puffer fish is 0.5mL:500g.
4. The hybridization method according to claim 1, wherein the water temperature of the nutrient enrichment culture is 26℃to 28 ℃; the salinity of the nutrition-enriched culture is 28-30; the time of the nutrient enrichment culture is 45-50 d.
5. The hybridization method according to claim 1, wherein the fertilized eggs are hatched at a seawater temperature of 25 to 26℃and a seawater salinity of 28 to 30.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310218875.8A CN115886159B (en) | 2023-03-09 | 2023-03-09 | Gonad development strengthening bait and hybridization method for puffer fish and puffer fish hexaflumuron |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310218875.8A CN115886159B (en) | 2023-03-09 | 2023-03-09 | Gonad development strengthening bait and hybridization method for puffer fish and puffer fish hexaflumuron |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115886159A CN115886159A (en) | 2023-04-04 |
| CN115886159B true CN115886159B (en) | 2023-10-20 |
Family
ID=86474911
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310218875.8A Active CN115886159B (en) | 2023-03-09 | 2023-03-09 | Gonad development strengthening bait and hybridization method for puffer fish and puffer fish hexaflumuron |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115886159B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101785440A (en) * | 2010-03-12 | 2010-07-28 | 河北省水产研究所 | Method for improving growth trait of Fugu bimaculatus |
| CN105075936A (en) * | 2015-06-19 | 2015-11-25 | 福建省水产研究所 | Method for quickly cultivating Takifugu bimaculatus |
| CN106852283A (en) * | 2016-12-28 | 2017-06-16 | 大连富泰水产养殖有限公司 | A kind of Fugu rubripes breeding method |
| CN108077116A (en) * | 2017-12-08 | 2018-05-29 | 中国水产科学研究院东海水产研究所 | A kind of artificial raise seedling method of wild six spots thorn Puffer |
| CN108077122A (en) * | 2017-12-25 | 2018-05-29 | 中国水产科学研究院东海水产研究所 | A kind of artificial acclimation method of wild lead point Fugu |
| CN109601438A (en) * | 2019-01-28 | 2019-04-12 | 南通龙洋水产有限公司 | A kind of Fugu rubripes raun and fugu obscurus milter cross breeding method |
| CN109819913A (en) * | 2019-03-26 | 2019-05-31 | 海南晨海水产有限公司 | A kind of artificial breeding method of Porcupinefiss |
| CN113317241A (en) * | 2021-06-09 | 2021-08-31 | 中国水产科学研究院东海水产研究所 | Method for gonadotropic development and artificial induced spawning of parent fish of takifugu hexamaculatus |
| CN114651751A (en) * | 2021-12-29 | 2022-06-24 | 广东省鳇鲸海洋生物科技有限公司 | Artificial fry breeding method for yellow lip fish |
-
2023
- 2023-03-09 CN CN202310218875.8A patent/CN115886159B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101785440A (en) * | 2010-03-12 | 2010-07-28 | 河北省水产研究所 | Method for improving growth trait of Fugu bimaculatus |
| CN105075936A (en) * | 2015-06-19 | 2015-11-25 | 福建省水产研究所 | Method for quickly cultivating Takifugu bimaculatus |
| CN106852283A (en) * | 2016-12-28 | 2017-06-16 | 大连富泰水产养殖有限公司 | A kind of Fugu rubripes breeding method |
| CN108077116A (en) * | 2017-12-08 | 2018-05-29 | 中国水产科学研究院东海水产研究所 | A kind of artificial raise seedling method of wild six spots thorn Puffer |
| CN108077122A (en) * | 2017-12-25 | 2018-05-29 | 中国水产科学研究院东海水产研究所 | A kind of artificial acclimation method of wild lead point Fugu |
| CN109601438A (en) * | 2019-01-28 | 2019-04-12 | 南通龙洋水产有限公司 | A kind of Fugu rubripes raun and fugu obscurus milter cross breeding method |
| CN109819913A (en) * | 2019-03-26 | 2019-05-31 | 海南晨海水产有限公司 | A kind of artificial breeding method of Porcupinefiss |
| CN113317241A (en) * | 2021-06-09 | 2021-08-31 | 中国水产科学研究院东海水产研究所 | Method for gonadotropic development and artificial induced spawning of parent fish of takifugu hexamaculatus |
| CN114651751A (en) * | 2021-12-29 | 2022-06-24 | 广东省鳇鲸海洋生物科技有限公司 | Artificial fry breeding method for yellow lip fish |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115886159A (en) | 2023-04-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100464633C (en) | Erythtoculer ilishaefornis and megajobrama amblycephala crossbreeding method | |
| CN101438690B (en) | Artificial propagation method of crocodile | |
| KR100991274B1 (en) | Culture method young crab for snow crab | |
| CN111956783B (en) | Artificial spawning induction mixture for largehead black weever and artificial breeding method | |
| CN107581106B (en) | A kind of artificial fecundation method of platax orbicularis | |
| CN101243778B (en) | Method for cultivating Protosalanx hyalocranius fingerlings | |
| CN111264423B (en) | Seedling raising method for acanthocephalus spinosus | |
| CN108967278A (en) | A kind of artificial fecundation method of Scatophagus argus (Linnaeus) | |
| CN113508766A (en) | Method for domesticating and feeding largemouth bass fry | |
| KR101169013B1 (en) | Method for mass producing artificial seed of Misgurnus sp. | |
| CN108770741A (en) | A kind of hatching method of east star spot | |
| CN101658146A (en) | Salt tolerant breeding technology for tilapia | |
| CN110612930A (en) | Artificial breeding method for rainbow trout | |
| CN104542478A (en) | Imitative ecological breeding and reproduction technology of andrias davidianus | |
| KR20140040336A (en) | Aquaculture method for a cynoglossus semilaevis | |
| CN102308769B (en) | Exopalaemon modestus fry culturing method through combining net cage with aquarium | |
| Ndimele et al. | Growth performance of the reciprocal hybrids of Clarias gariepinus (Burchell, 1822) and Heterobranchus bidorsalis (Valenciennes, 1840) | |
| CN115886159B (en) | Gonad development strengthening bait and hybridization method for puffer fish and puffer fish hexaflumuron | |
| Chellam | Biology of pearl oyster Pinctada fucata (Gould) | |
| CN102792910A (en) | Cultivating method of oncorhynchus mykiss walbaum gynoecial hybrid offspring seed | |
| CN102812913A (en) | Cultivation method for hybridized all-female golden trout fries | |
| CN110074019A (en) | A kind of fugu obscurus, knife long-tailed anchovy and the cultivation of soft-shelled turtle stereo ecological method | |
| KR101289847B1 (en) | New culturing method of penshell pearl | |
| CN109169416A (en) | Pelteobagrus fulvidraco offspring seed cultivation method | |
| KR101611139B1 (en) | the feed composition and Hatchery technique for anguillidae |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |