CN108522371B - Hatching method for dormant eggs of black pearl killifish - Google Patents
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- 235000013601 eggs Nutrition 0.000 title claims abstract description 140
- 230000012447 hatching Effects 0.000 title claims abstract description 92
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 54
- 238000011534 incubation Methods 0.000 claims abstract description 47
- 238000005507 spraying Methods 0.000 claims abstract description 37
- 241000251468 Actinopterygii Species 0.000 claims abstract description 32
- 241000276569 Oryzias latipes Species 0.000 claims abstract description 22
- 230000018109 developmental process Effects 0.000 claims abstract description 17
- 230000000284 resting effect Effects 0.000 claims abstract description 9
- 239000003814 drug Substances 0.000 claims abstract description 7
- 239000002184 metal Substances 0.000 claims description 32
- 239000002689 soil Substances 0.000 claims description 17
- 239000003415 peat Substances 0.000 claims description 16
- 229920000742 Cotton Polymers 0.000 claims description 15
- 238000007789 sealing Methods 0.000 claims description 14
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 11
- 241001263038 Viguiera Species 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 11
- 241001137324 Cyprinodontiformes Species 0.000 claims description 7
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 claims description 6
- 229960000907 methylthioninium chloride Drugs 0.000 claims description 6
- 238000003860 storage Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 238000002372 labelling Methods 0.000 claims description 5
- 239000003755 preservative agent Substances 0.000 claims description 5
- 230000002335 preservative effect Effects 0.000 claims description 5
- 238000005096 rolling process Methods 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 4
- 238000004321 preservation Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 3
- 238000010438 heat treatment Methods 0.000 abstract description 3
- 238000009395 breeding Methods 0.000 description 6
- 230000001488 breeding effect Effects 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 5
- 102000002322 Egg Proteins Human genes 0.000 description 5
- 108010000912 Egg Proteins Proteins 0.000 description 5
- 235000014103 egg white Nutrition 0.000 description 5
- 210000000969 egg white Anatomy 0.000 description 5
- 230000017448 oviposition Effects 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000003889 eye drop Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
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- 241000243321 Cnidaria Species 0.000 description 1
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- 244000052616 bacterial pathogen Species 0.000 description 1
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Classifications
-
- 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
- A01K61/10—Culture of aquatic animals of fish
- A01K61/17—Hatching, e.g. incubators
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention relates to a hatching method and discloses a method for hatching dormant eggs of black pearl medaka, which comprises the following steps: 1) storing the resting eggs; 2) pretreating dormant eggs; 3) hatching in water; 4) hatching the un-hatched eggs. According to the method, the fertilized eggs are stored in a soilless manner, the development condition of the fish eggs can be accurately judged, the incubation time node is more accurate, the development process is accelerated by heating and spraying medicines, the incubation rate is improved by 30% compared with the traditional incubation method, the incubation frequency is only required to be 2 times, the incubation time is greatly shortened, manual egg picking is not needed, the manpower is greatly saved, and the method is suitable for incubation of the dormant eggs of the black pearl killifish for commercial application.
Description
Technical Field
The invention relates to a hatching method, in particular to a hatching method for dormant eggs of black pearl killifishes.
Background
The black pearl killifish belongs to oviposition killifish grown in one year in south America, has extremely attractive body color, can be said to be one of few fish species which can be compared favorably with seawater coral reef fish in fresh water ornamental fish, and is lively and lively in a character by virtue of the small body type, so that the black pearl killifish is naturally and absolutely matched when placed in a waterweed jar. In addition, most of the medaka eggs have wide adaptability to water quality and are easy to breed, and the characteristics make the medaka eggs become one of popular tropical ornamental fish species at home and abroad. In particular, fertilized eggs of medaka have a characteristic of resting for a certain period of time, and therefore, the medaka and the fish eggs can be sold on the market. However, medaka oviposits are extremely specific in terms of reproductive behavior: the method has the advantages of multiple oviposition, uncertain time, small amount (more oviposition at one time, dozens of eggs, and one or two eggs at a time), short fish egg storage time, and only 12-52% of natural killifish eggs sold on the market after one month. These reasons all cause that the yield of the natural oviparous medaka is extremely low, and some species are rare endangered species in the world. However, the market industrialization of killifishes has a plurality of problems of small spawning amount of breeding fishes, low fertilization rate, low egg hatching rate, low fry breeding survival rate and the like, and the development and the industrial development of the ornamental type egg-grown killifishes market are restricted.
Chinese patent application No. 201610259750.X discloses a method for efficient breeding and hatching of medaka, which provides a method for efficient breeding and hatching of medaka with high breeding controllability and high breeding efficiency, but the method is directed to medaka, which and black pearl medaka in different families and genera in classification, after the medaka eggs are mature, the medaka produces the medaka without leaving the mother, but the medaka is hung outside a maternal cloaca through a sticky wire, has no dormant period, is easy to hatch, and in the case of one-year medaka hatching, the fertilized eggs must go to dormancy after leaving water for 2-3 months, and even in the case of the same batch of fertilized eggs, the hatching time is not completely consistent, so that the method is not suitable for commercialization.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for hatching dormant eggs of black pearl killifishes, fertilized eggs are stored in a soilless mode, the development process is accelerated through temperature rise and medicine spraying, the hatching rate is greatly improved, the cost is reduced, and the method is suitable for commercial application.
The specific technical scheme of the invention is as follows: the method comprises the following steps:
1) and (3) preservation of dormant eggs: storing the dormant eggs in a soilless mode, and checking the development condition of the eggs after the fish eggs are dormant for 50 days at the temperature of 23-27 ℃;
2) pretreatment of resting eggs: when dormant eggs have golden eye circles, increasing the storage temperature by 1-2 ℃, taking out the dormant eggs and placing the dormant eggs in a metal screen when all dormant eggs have golden eye circles for 14-16 days, and spraying hatching water on the eggs;
3) hatching in water: after spraying is finished for 18-22min, placing the metal screen mesh into an incubation cylinder, adding 13-17cm of incubation water into the incubation cylinder, enabling the upper end of the metal screen mesh to be submerged for 2-4cm, hatching partial eggs after 5.5-6.5h, and inclining a screen mesh opening to one side to enable the larvae which are not swim out to be free from the screen mesh;
4) hatching the unhatched eggs: washing the unhatched eggs with clear water, draining, placing on egg-carrying material, storing at 27-28 deg.C for 10-12 days, spraying the medicinal liquid, and incubating in water.
The eggs are usually stored in peat soil in the traditional hatching mode, after the eggs are hatched for the first time, the eggs containing the un-hatched eggs and the peat soil are taken out together, drained and sucked by newspaper, so that the operation is complicated, the hatching rate is low, and the ratio of the hatched larval fish lying on the bottom is high; because the eggs are mixed in the soil, the development condition of the eggs is inconvenient to observe, the optimal initial hatching time cannot be found accurately, and a small amount of deciduate larvae after initial hatching die due to the fact that the larvae cannot dissociate from a peat soil layer and affect the eggs for secondary hatching, the traditional hatching mode usually needs three hatching or four hatching to enable the hatching rate to reach 60%; according to the invention, the dormant eggs are stored in a soilless manner, so that the development condition of the fish eggs can be accurately judged, and the incubation time node is more accurate.
The dormant eggs of the black pearl medaka are developed in 5 stages, and the 5 stages can be distinguished from color change and are respectively as follows: the method is characterized by comprising the steps of transparency, light yellow, deep yellow, black eyepoint and golden eyedrop, aiming at different requirements on temperature in the process of the growth of the black pearl killifish, and accelerating the growth process of the fish eggs by heating when the golden eyedrop appears.
The invention can stimulate the fish eggs to be stripped through spraying and medicine spraying, reduce the hatching times and improve the hatching rate by 30 percent.
Preferably, the soilless mode in the step 1) is to seal the dormant eggs in a sealing bag wrapped by biochemical cotton and preservative films.
Preferably, the development of the eggs is checked every 2 to 3 days in step 1).
Preferably, the preparation method of the water for hatching in the step 2) comprises the following steps: adding 9-11wt% of peat soil leachate and 20-30ppm of glutaraldehyde into water at the temperature of 26.5-27.5 ℃, and circularly filtering for 3 days by using a biochemical cotton filter.
When the black pearl killifish is hatched, weak acid water quality and light brown water color are needed, peat soil leachate is added and filtered, so that not only can the water quality be adjusted, but also other impurities and germs can be prevented from being introduced due to the direct addition of peat soil.
Preferably, the spraying method in the step 2) comprises the following steps: placing the metal screen above water surface of the hatching jar, spraying for 4.5-5.5min, and spraying for 4.5-5.5min at intervals of 18-22 min.
Preferably, the hatching jar in the step 3) has a length, width and height (45-55) cm x (25-35) cm x (20-30) cm.
Preferably, the egg-carrying material in the step 4) is black compact biochemical cotton which is soaked and squeezed in peat soil leachate containing 4-6ppm of methylene blue.
Methylene blue can prevent the fish eggs which are not completely developed from infecting Saprolegnia.
Preferably, the method for preserving the unhatched eggs in the step 4) comprises pouring the eggs on the egg-carrying material, rolling up the egg-carrying material, sealing the bag in a fresh-keeping bag, attaching a label to the bag, and noting the fish species, the initial incubation date and the number of eggs.
Preferably, the drug spraying method in the step 4) is spraying with 80-120ppm glutaraldehyde for 2-4 min.
Preferably, the method for hatching the water in the step 4) comprises the steps of placing a metal screen into a hatching cylinder after spraying is finished for 10min, adding 13-17cm of hatching water into the hatching cylinder, and placing the upper end of the metal screen 2-4cm below the hatching water, and hatching for 22-26h to finish, wherein eggs which cannot be hatched or hatched larvae on the bottom can be eliminated.
The glutaraldehyde with higher concentration can effectively stimulate the fish egg to be stripped, and reduce the hatching times.
Compared with the prior art, the invention has the beneficial effects that: according to the method, the fertilized eggs are stored in a soilless manner, the development condition of the fish eggs can be accurately judged, the incubation time node is more accurate, the development process is accelerated by heating and spraying medicines, the incubation rate is improved by 30% compared with the traditional incubation method, the incubation frequency is only required to be 2 times, the incubation time is greatly shortened, manual egg picking is not needed, the manpower is greatly saved, and the method is suitable for incubation of the dormant eggs of the black pearl killifish for commercial application.
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1
Preparing hatching water: adding 10 wt% of peat soil leachate and 20ppm of glutaraldehyde into water at the temperature of 27 ℃, and circularly filtering for 3 days by using a biochemical cotton filter.
Sealing the dormant eggs in a sealing bag wrapped by biochemical cotton and preservative film, and checking the development condition of the eggs every 3 days after the fish eggs are dormant at 25 ℃ for 50 days; when dormant eggs have golden eye circles, increasing the storage temperature by 2 ℃, taking out the dormant eggs and placing the dormant eggs in a metal screen when all dormant eggs have golden eye circles on day 15, spraying the eggs with hatching water for 5min, and spraying for 5min at intervals of 20 min; after spraying is finished for 20min, placing the metal screen into an incubation cylinder with the specification of length, width and height of 50cm multiplied by 30cm multiplied by 25cm, adding 15cm of incubation water into the incubation cylinder, enabling the upper end of the metal screen to be submerged for 3cm, hatching partial eggs after 6h, and inclining a screen opening to one side to enable the larvae which are not swim out to be free from the screen; washing the un-incubated egg white with water, draining, soaking in peat soil leachate containing 5ppm methylene blue, squeezing to obtain black compact biochemical cotton, rolling up the egg-carrying material, sealing in a fresh-keeping bag, labeling, and indicating the fish species, the initial incubation date and the number of eggs; storing at 27 deg.C for 11 days, spraying with 100ppm glutaraldehyde for 3min, placing metal screen in hatching tank after spraying for 10min, adding hatching water 15cm into the hatching tank, leaving the upper end of the metal screen 3cm below the hatching water, and hatching for 24h to remove un-hatched eggs or hatched larva fish.
Example 2
Preparing hatching water: 9 wt% of peat soil leachate and 30ppm of glutaraldehyde are added into water, the water temperature is 26.5 ℃, and the mixture is circularly filtered for 3 days by a biochemical cotton filter.
Sealing the dormant eggs in a sealing bag wrapped by biochemical cotton and preservative film, and checking the development condition of the eggs every 2 days after the fish eggs are dormant at 27 ℃ for 50 days; when dormant eggs have golden eye circles, increasing the storage temperature by 1 deg.C, and when all dormant eggs have golden eye circles, taking out the dormant eggs, placing in a metal screen, spraying with hatching water for 4.5min, and spraying for 4.5min every 22 min; 22min after spraying, placing a metal screen into an incubation cylinder with the specification of length, width and height of 55cm multiplied by 35cm multiplied by 20cm, adding 13cm of incubation water into the incubation cylinder, enabling the upper end of the metal screen to be submerged for 2cm, hatching out partial eggs after 6.5h, and inclining a screen opening to one side to enable the fry which are not swim out to be free from the screen; washing the un-incubated egg white with water, draining, soaking in peat soil leachate containing 4ppm methylene blue, squeezing to obtain black compact biochemical cotton, rolling up the egg-carrying material, sealing in a fresh-keeping bag, labeling, and indicating the fish species, the initial incubation date and the number of eggs; storing at 27 deg.C for 12 days, spraying with 120ppm glutaraldehyde for 2min, placing metal screen in hatching tank after spraying for 10min, adding hatching water 13cm into the hatching tank, leaving the upper end of the metal screen 2cm below the hatching water, hatching for 26h, and eliminating un-hatched eggs or hatched larva fish.
Example 3
Preparing hatching water: adding 11wt% of peat soil leachate and 25ppm of glutaraldehyde into water at the temperature of 27.5 ℃, and circularly filtering for 3 days by using a biochemical cotton filter.
Sealing the dormant eggs in a sealing bag wrapped by biochemical cotton and a preservative film, and checking the development condition of the eggs every 3 days after the fish eggs are dormant for 50 days at 23 ℃; when the dormant eggs have golden eye circles, increasing the storage temperature by 2 ℃, taking out the dormant eggs and placing the dormant eggs in a metal screen when all the dormant eggs have golden eye circles on the 16 th day, spraying the eggs with hatching water for 5.5min, and spraying for 5.5min at intervals of 18 min; after spraying for 18min, placing the metal screen into an incubation cylinder with the specification of length, width and height of 45cm multiplied by 25cm multiplied by 30cm, adding 17cm of incubation water into the incubation cylinder, immersing the upper end of the metal screen in water for 4cm, hatching partial eggs after 5.5h, and inclining a screen opening to one side to enable the larvae which are not swim out to be free from the screen; washing the un-incubated egg white with water, draining, soaking in peat soil leachate containing 6ppm methylene blue, squeezing to obtain black compact biochemical cotton, rolling up the egg-carrying material, sealing in a fresh-keeping bag, labeling, and indicating the fish species, the initial incubation date and the number of eggs; storing at 28 deg.C for 10 days, spraying with 80ppm glutaraldehyde for 4min, placing metal screen in hatching tank after spraying for 10min, adding 17cm hatching water into the hatching tank, leaving the upper end of the metal screen 4cm below the hatching water, and hatching for 22h to remove un-hatched eggs or hatched larva fish.
Example 4
The incubation method was completely identical to that of example 1, except that the resting temperature of the eggs was 23 ℃.
Example 5
The incubation method was completely identical to that of example 1, except that the resting temperature of the eggs was 27 ℃.
Comparative example 1
The incubation water was prepared in the same manner as in example 1.
Storing the resting eggs in peat soil, and checking the development condition of the eggs every 3 days after the fish eggs are dormant at 23 ℃ for 50 days; on the 15 th day after all the dormant eggs have golden eye circles, taking out the dormant eggs, placing the dormant eggs in a metal screen mesh, placing the metal screen mesh in an incubation cylinder with the specification of length, width and height of 50cm multiplied by 30cm multiplied by 25cm, adding 15cm of incubation water into the incubation cylinder, enabling the upper end of the metal screen mesh to be submerged by 3cm, hatching part of the eggs after 6 hours, and inclining a screen mesh port to one side to enable the fry which are not swim out to be free from the screen mesh; washing the un-incubated egg white with water, draining, recovering the un-incubated egg white, sealing in a fresh-keeping bag, and labeling with the fish species, the initial incubation date and the number of eggs; after being preserved for 11 days at the temperature of 27 ℃, the metal screen is placed in the original hatching jar again for hatching, after 24 hours, the unhatched eggs are recovered and preserved again, and the hatching steps are repeated for 4 times.
Comparative example 2
The incubation method was completely identical to comparative example 1, except that the resting temperature of the eggs was 25 ℃.
Comparative example 3
The incubation method was completely identical to comparative example 1, except that the resting temperature of the eggs was 27 ℃.
Selecting 600 black pearl medaka eggs of the same day age, dividing the eggs into two groups of 6 parts, hatching 3 parts of 1 group of the eggs in a comparative example 1-3 mode for 100 eggs in each part, hatching 4 times in each part, hatching 3 parts of 2 groups of the eggs in examples 1, 4 and 5 modes for 2 times in each part, and the hatching results are shown in table 1.
Table 1: results of comparison tests of traditional fish egg preservation and hatching modes of black pearl killifishes and modes adopted by the patent
According to the table 1, the hatching method has the hatching times half that of the traditional method, the total hatching rate is improved by about 30% compared with the traditional method, the survival rate of the fries is improved by more than 10%, the hatching success rate of dormant eggs of the black pearl killifishes is remarkably improved, the hatching time and the labor cost are saved, and the hatching method is beneficial to commercialization.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (5)
1. A method for hatching dormant eggs of black pearl killifishes is characterized by comprising the following steps: the method comprises the following steps:
1) and (3) preservation of dormant eggs: storing the dormant eggs in a soilless manner, and after the fish eggs are dormant at the temperature of 23-27 ℃ for 50 days, checking the development condition of the eggs, wherein the soilless manner is to seal the dormant eggs in a sealing bag wrapped by biochemical cotton and preservative films;
2) pretreatment of resting eggs: when dormant eggs have golden eye circles, increasing the storage temperature by 1-2 ℃, taking out the dormant eggs and placing the dormant eggs in a metal screen when all dormant eggs have golden eye circles for 14-16 days, and spraying hatching water on the eggs;
the preparation method of the hatching water comprises the following steps: adding 9-11wt% of peat soil leachate and 20-30ppm of glutaraldehyde into water at the temperature of 26.5-27.5 ℃, and circularly filtering for 3 days by using a biochemical cotton filter;
3) hatching in water: after spraying is finished for 18-22min, placing the metal screen mesh into an incubation cylinder, adding 13-17cm of incubation water into the incubation cylinder, enabling the upper end of the metal screen mesh to be submerged for 2-4cm, hatching partial eggs after 5.5-6.5h, and inclining a screen mesh opening to one side to enable the uniparted fries to free the metal screen mesh;
4) hatching the unhatched eggs: washing unhatched eggs with clear water, draining, placing on an egg carrying material, preserving at 27-28 ℃ for 10-12 days, spraying the medicine, and incubating in water, wherein the incubation method in water comprises the steps of after spraying is finished for 10min, placing a metal screen into an incubation cylinder, adding 13-17cm of incubation water into the incubation cylinder, immersing the upper end of the metal screen in the incubation water for 2-4cm, and incubating for 22-26h to finish, so that eggs which cannot be incubated or young fish lying on the bottom after incubation can be eliminated;
the egg-carrying material is black compact biochemical cotton which is soaked and squeezed in peat soil leachate containing 4-6ppm of methylene blue;
the medicine spraying method in the step 4) is to spray 80-120ppm glutaraldehyde for 2-4 min.
2. The method for hatching dormant eggs of black pearl medaka according to claim 1, wherein the development of the eggs is checked every 2 to 3 days in step 1).
3. The hatching method of dormant eggs of black pearl medaka according to claim 1, wherein the spraying method in the step 2) is: placing the metal screen above water surface of the hatching jar, spraying for 4.5-5.5min, and spraying for 4.5-5.5min at intervals of 18-22 min.
4. The method for hatching the dormant eggs of black pearl medaka according to claim 1, wherein the size of the hatching jar in the step 3) is length, width and height (45-55) cm x (25-35) cm x (20-30) cm.
5. The method for hatching the dormant eggs of black pearl medaka according to claim 1, wherein the unhatched eggs in the step 4) are stored by pouring the eggs on an egg carrying material, rolling up the egg carrying material, sealing and labeling a fresh keeping bag, and noting the fish species, the initial hatching date and the number of eggs.
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